WO2010128163A2 - Small molecule inhibitors of influenza a and b virus and respiratory syncytial virus replication - Google Patents

Small molecule inhibitors of influenza a and b virus and respiratory syncytial virus replication Download PDF

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WO2010128163A2
WO2010128163A2 PCT/EP2010/056304 EP2010056304W WO2010128163A2 WO 2010128163 A2 WO2010128163 A2 WO 2010128163A2 EP 2010056304 W EP2010056304 W EP 2010056304W WO 2010128163 A2 WO2010128163 A2 WO 2010128163A2
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Prior art keywords
methyl
thiazole
thienyl
thiophen
carboxylic acid
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PCT/EP2010/056304
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French (fr)
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WO2010128163A3 (en
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Ulrich Kessler
Charlène RANADHEERA
Muriel Joubert
Bruno Giethlen
Fabrice Garrido
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Pike Pharma Gmbh
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Priority to EP10719757A priority Critical patent/EP2427447A2/en
Publication of WO2010128163A2 publication Critical patent/WO2010128163A2/en
Publication of WO2010128163A3 publication Critical patent/WO2010128163A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
    • C07D333/22—Radicals substituted by doubly bound hetero atoms, or by two hetero atoms other than halogen singly bound to the same carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • the present invention relates to small molecules inhibiting the replication of influenza A and B virus and respiratory syncytial virus (RSV), and the use of such compounds for treating influenza A and B and RSV infections, in humans, mammals and birds.
  • RSV respiratory syncytial virus
  • Influenza viruses are negative-stranded RNA viruses that cause yearly epidemics as well as recurring pandemics, resulting in high numbers of human cases and severe economic burden.
  • pandemic influenza A viruses such as the 1 91 8 "Spanish" flu or H5N 1
  • pandemic influenza B viruses contribute greatly to the annual recurring epidemics that cause the vast majority of human cases and medical cost.
  • the WHO recommends an annual vaccination against circulating influenza A (FIuA) and B (FIuB) strains.
  • FIuA circulating influenza A
  • FIuB circulating influenza A
  • current vaccines confer incomplete protection against epidemic influenza.
  • oseltamivir oseltamivir
  • RelenzaTM zanamivir
  • RSV Human respiratory syncytial virus
  • SSV Human respiratory syncytial virus
  • Pa-ramyxoviridae RNA virus of the family Pa- ramyxoviridae
  • Treatment is mainly limited to supportive care, including oxygen.
  • Palivizumab (SynagisTM) is used as a prophylactic drug in prevention of respiratory RSV infections for infants with a high risk of infection.
  • Ribavirin has been used for treating RSV infections, but showed limited effectiveness.
  • One object of the invention is to provide new, improved and/or alternative influenza and RSV antiviral com pounds.
  • Another object of the invention is to obviate or mitigate disadvantages of influenza antiviral agents and RSV antiviral agents known from the state of the art.
  • Advantageous embodi- ments are given in the dependent claims.
  • a compound according to the invention is 2-(3-Q 5 4-Q 4 -5-Q 3 -thien-2-yl)4-R 3 -5-R 2 -l ,3-thiazol, 2-(3-Q 5 4-Q 4 - 5-Q 3 -fur-2-yl)-4-R 3 -5-R 2 -l ,3-thiazol, 2-(3-Q 5 4-Q 4 -5-Q 3 -thien-2-yl)-5-R 2 -l ,3,4-thiadiazol, 2-(3-Q 5 -4-Q 4 -5-Q 3 -fur- 2-yl)-5-R 2 -l ,3,4-thiadiazol, 2-(2-Q 3 -4-Q 5 -l ,3-thiazol-5-yl)4-R 3 -5-R 2 -l ,3-thiazol, 2-(2-Q 3 4-Q 5 -l ,3-oxazol-5-yl)4- R 3 -5-R 2 -l ,
  • R 2 is selected from a group consisting of: hydrogen, cyano, formyl, acetyl, ethylcarbonyl, propylcarbonyl, isopropylcarbonyl, carboxy, carboxymethyl, methoxycarbonyl, eth- oxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert- butoxycarbonyl, methoxycarbonylmethyl, methoxycarbonylethyl, ethoxycarbonylmethyl, methoxycarbon- ylethyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trihalogenomethyl, difluoromethyl, dihalogenomethyl, fluoromethyl, halogenomethyl, phenyl, 1 -naphthyl, 2-nap
  • Said compounds according to the invention are particularly advantageous as a medicament.
  • the above defined compounds according to the invention are particular advantageous for treating and/or preventing influenza type A and/or influenza type B infections in humans, mammals and/or birds; as well as for treating and/or preventing respiratory syncytial virus (RSV) infections in humans, mammals and/or birds.
  • RSV respiratory syncytial virus
  • the compound according to the invention can be used for the manufacture of a medicament for the treatment and/or prevention of influenza type A and/or influenza type B infections in humans, mam- mals and/or birds, and/or for the treatment and/or prevention of respiratory syncytial virus infections in humans, mammals and/or birds.
  • a pharmaceutical composition according to the invention comprises a compound according to the invention.
  • Advantageously such a composition comprises one or more excipients.
  • a method for identifying small molecules and/or peptidic inhibitors or dis- ruptors for protein-protein interactions comprises the steps of: a) selection of a proteome of interest; b) identification of potential target regions of PPI inhibitors; c) detection of drugable protein- binding domains; d) synthesis of peptides corresponding to the drugable protein-binding domains; e) testing of the synthesized peptides for their ability to bind the protein partner involved in the PPI; f) development of a binding assay for the identification of small molecules and/or peptidic inhibitors or disrupters based on the positively tested peptides; and g) high-throughput screening with the binding assay and selecting the active small molecules and/or peptidic inhibitors or disruptors.
  • compounds in accordance with the present invention are able to inhibit protein-protein interaction of the PA and PBl subunits of the heterotrimeric viral RNA polymerase complex of both influenza virus types A and B, and thus are able to inhibit replication of influenza A and B virus.
  • the viral polymerase subunit interaction domain turned out as an effective target for the new antiviral compounds, since correct assembly of the three viral polymerase subunits PBl , PB2 and PA is required for viral RNA synthesis and infectivity. Structural data for the entire trimeric complex is missing.
  • the crucial PA interaction domain of PBl consists of a 3io-helix formed by amino acids (amino acids 5-1 1 ).
  • the domain is highly conserved and virus type specific among both, influenza A and B viruses.
  • an Enzyme-Linked Immunosorbent Assay (ELISA) based screening assay and other assays are used to prescreen compounds according to the invention that show antiviral activity against influenza A and B viruses. Since they are effective against both virus types, such compounds represent an attractive alternative to neuraminidase inhibitors. Therefore, the present invention represents a major step toward a sorely needed, near-universal medicament against influenza virus, and one which, due to its protein- protein interaction domain target, will likely be less susceptible to the emergence of drug-resistant strains for which influenza is well known.
  • RSV respiratory syncytial virus
  • the compounds according to the invention can be used as a medicament, particularly as an influenza virus and/or RSV replication inhibitor and an influenza and/or RSV preventive/therapeutic agent, respectively.
  • Influenza Type A and B Therapeutic target
  • amino acids are preferably indicated by the IUPAC one letter code in the present application. Whenever three letter codes are used, they are also in accordance with IUPAC.
  • the letter X is used to indicate a wildcard/variable or other amino acid at a certain position.
  • Table Ia shows the inhibitory concentrations of FluA/FluB-derived peptides determined by competitive ELISA. Competitor peptides (0.048 to 300OnM) were mixed with cell extracts containing HA-tagged PA from either FIuA or FIuB. Table 1 lists 12 competitive peptides. The first peptide PBIM 5 A is the FIuA wild type, the second row shows the FIuB wild type. For the peptides of rows 3 to 8 letters indicate FIuB specific amino acids. Rows 9 to 12 list further competitive peptides with amino acids at position 6 being neither FIuA nor FIuB specific. Standard deviation is indicated in parenthesis. Asterisks indicate highest concentrations of peptides used without reaching 50% inhibition.
  • Table 1 c the amino acid sequences at amino acid residues X 5 to Xi 0 of wild type A mutants are indicated. Said peptides exhibit lower activities than the above mentioned peptides according to Tables I a and 1 b.
  • Table I c qualitative overview of further peptides
  • the synthesized or isolated influenza virus replication-inhibiting peptides interacting with the inhibition target for the small molecules compounds according to the invention comprise an amino acid sequence Of X 5 X 6 X 7 X 8 X 9 XiO, wherein X 5 is P; X 6 is T, Y, F, W, H, C, I, L, V, A or M; X 7 is L or F; X 8 is L, I, F or M; X 9 is F, Y, W, H, L, R or S, and Xio is L, I or Y.
  • Said amino acid sequence is at least 60 %, preferably at least 70%, more preferably at least 80% or 90% identical to the polypeptide according to the wild type PBI mA which is M DVN PTLLFLK.
  • those peptides are preferred which comprising the amino acid sequence Of X 6 X 7 X 8 XgXi 0 , wherein X 6 is T, Y, F, W, H, C, I, L or V; X 7 is L or F; X 8 is L or I; X 9 is F, Y or W and Xi 0 is L.
  • Even more preferred according to certain embodiments are peptides that comprise the amino acid sequence Of X 6 X 7 , wherein X 6 is T, Y, F, W, H, C, I, L or V and X 7 is L or F.
  • Effective peptides advantageously comprise at least 1 1 residues Xi n, whereby preferably the proteins comprise the amino acid sequence M DVN PX6X7 LFLKVPAQ wherein X6 is selected from the group: T, Y, F, W, H. C, A, I, L, V or M and X7 is selected from the group L or F.
  • a preferred peptide comprises an amino acid sequence elected from the group: MDVNPYFLFLKVPAQ, MDVNPYLLFLKVPAQ, MDVNPWLLFLKVPAQ or MDVNPFLLFLKVPAQ.
  • the peptides com prise at least 1 5 residues Xi 1 5 according to the wild type PBl 1 1 5 A but not the wild type sequence MDVNPTLLFLKVPAQ.
  • Table 2 shows the 50%-inhibitory concentrations (IC 50 ) of FluA-derived PBl peptides determined by competitive ELISA.
  • Peptide PBI 1 25 A was immobilized on microwell plates and incubated with increasing concentrations of competitor peptides and cell extract containing HA-tagged PA of FIuA. Bound PA was detected by HA-specific antibodies as described above. Standard deviation is shown in parenthesis. Asterisks indicate highest concentrations of peptides used without detectable inhibitory effect.
  • Grey boxes highlight amino acids that are part of the 3i O -helix, which comprises the core PA-binding region of PBl . Amino acids known to form hydrogen bonds with PA residues are represented in bold.
  • Table 3 illustrates the inhibitory concentrations (IC 50 ) of FluA-derived competitor peptides determined by ELISA.
  • Peptide PBl 1 . 25 A was again immobilized on microwell plates and incubated with increasing concentrations of competitor peptide and cell extract containing HA-tagged PA of FIuA. HA-specific antibodies detected bound PA. Standard deviations are shown in parenthesis. Asterisks indicate highest concentrations of peptides used without detectable inhibitory effect.
  • Figure 1 shows binding and inhibitory activity of PB1 1 -25AT6Y
  • Figure 1 shows binding and inhibitory activity of peptides binding to the inhibition target with a focus on the preferred protein PBl i 25 A 16Y shall be illustrated in the following part of the description
  • Figure I a shows in the upper panel the alignment of the consensus sequence of the N-terminal 25 ammo acids of FIuA and FIuB PBl , wherein the dotted box indicates the 3io-hel ⁇ x comprising the core PA-binding domain of PBl and the FluA-specific and FluB-specific ammo acids are printed in bold letters
  • Middle and lower panels show the alignment of the N-terminal 25 ammo acids of all available FIuA and FIuB sequences derived from PBl full length sequences provided by the NCBI influenza virus database
  • Figure I c provides graphic information on the structure of FIuA PBI i i s bound to FIuA PA T6 forms a hydrogen bond to a water molecule Molecular modeling suggests that the aromatic side chain in the mutant T6Y fits into a hydrophobic pocket and displaces the water molecule
  • the polymerase inhibitory activity of PBI i 25 -der ⁇ ved CFP fusion proteins in FIuA and FIuB polymerase reconstitution assays is shown in Figure I d
  • the activity in experiments containing all viral plasmids and Flag-CFP was set to 1 00%
  • Figure I e shows a plaque reduction assay using PBl i 25 A-Tat, PBl 1 25 A T6 ⁇ -Tat, PX-Tat (control peptide) with FIuA, FIuB and VSV (vesicular stomatitis virus)
  • PBl i 25 B-Tat could not be tested due to insolubility Error bars represent standard deviations
  • Figure 2a shows A/SC35M- and B/Yamagata/73-der ⁇ ved PBl chimeras used in tests according to Figure 2 b
  • Figure 2b shows human 293T cells which were transfected with expression plasmids coding for the indicated PBl proteins and the C-terminally hexahistidme- tagged PA of FIuA (FluAPA H
  • Cell lysates were prepared 24 hours post transfection and subjected to immunoprecipitation (IP) using ant ⁇ -HA (aHA) agarose
  • IP immunoprecipitation
  • aHA aHA
  • IP immunoprecipitation
  • aHA ant ⁇ -HA
  • FIG. 4a shows CFP-PBl fusion proteins used in tests according to Figure 4b
  • the complex formation of PBl 1 25 -der ⁇ ved CFP fusion proteins and HA-tagged PA of FIuA and FIuB is shown in Figure 4b
  • Indicated proteins were expressed in human 293T cells and binding of the CFP fusion proteins was analyzed by immunoprecitation (IP) of PA using anti-HA agarose and subsequent immunoblotting (IB) Precipitated material was analyzed by Western blot using the indicated antibodies for the presence of either HA- tagged PA or CFP Molecular weights are shown in kilodaltons
  • Virus strains For the infection experiments A/WSN/33 (H l N l ) according to Chanem et al (2007) and A/Tha ⁇ land/l (Kan-l )/2004 according to Chockephaibulkit et al (2005), B/Yamagat/73 according to Norton (1 987) and VSV (serotype Indiana) as described in Schwem mle (1 995) were used
  • Plasmid constructions Plasmids pCA-Flag-CFP and pCA-PBl 1 25 A-CFP, pCA-PBl -HA, the FIuA minireph- con plasmids and the expression plasmids for the FIuB minireplicon are described in Chanem (2007), Mayer (2007) and Pleschka (1 996)
  • the FIuB minigenome expression plasmid, pPoll-lucRT_B was obtained by cloning the firefly luciferase ORF (inverse orientation) flanked by the non-coding region of the segment 8 of the B/Yamagata/73 into the Sapl-digested plasmid pPoll-Sapl-Rib according to Pleschka (1 996)
  • a linker containing the first 25 codons of PBl was cloned into the EcoRI/
  • the A/B-chimeric expression plasmids were obtained by assembly PCR using the pCAPBl -HA plasmids of SC35M and B/Yamagata/73 and by cloning the resulting PCR product in pCA-PBl B / Ya m agata / 73 -HA digested with EcoRI/EcoRV.
  • H EK293T cells were transiently transfected with a plasmid mixture containing either FIuA- or FluB-derived PBl -, PB2-, PA- and NP-expression plasmids, polymerase I (Pol l)-driven plasmid transcribing an influenza A or influenza B virus-like RNA coding for the reporter protein firefly luciferase to monitor viral polymerase activity and with expression plasmids coding for the indicated CFP fusion proteins.
  • Both minigenome RNAs were flanked by non- coding sequences of segment 8 of FIuA and FIuB, respectively.
  • the transfection mixture also contained a plasmid constitutively expressing Renilla luciferase, which served to normalize variation in transfection efficiency.
  • the reporter activity was determined 24h post transfection and normalized using the Dual- Clu® Lufierase Assay System (Promega). The activity observed with transfection reactions containing Flag-CFP were set to 1 00%.
  • Peptide synthesis The solid-phase synthesis of the peptides was carried out on a Pioneer automatic peptide synthesizer (Applied Biosystems, Foster City, USA) em ploying Fmoc chemistry with
  • H EK293T cells were transfected with the indicated plasmids in 6- well plates using Metafectene (Biontex, Martinsried, Germany). Cells were incubated 24h post transfec- tion with lysis buffer (2OmM Tris pH7.5, 1 0OmM NaCI, 0.5mM EDTA, 0.5% NP40, 1 % Protease inhibitor Mix G, (Serva, Heidelberg, Germany), I mM DTT) for 1 5 min on ice. After centrifugation by 1 3.000 rpm at 4°C supernatant was incubated with anti HA-specific antibodies coupled to agarose beads (Sigma) for 1 h at 4°C.
  • lysis buffer 2OmM Tris pH7.5, 1 0OmM NaCI, 0.5mM EDTA, 0.5% NP40, 1 % Protease inhibitor Mix G, (Serva, Heidelberg, Germany), I mM DTT
  • Plaque reduction assay The experiments were carried out as described by Schmidke (2001 ) with modifications. Confluent MDCK cells were infected with I OOPFU of A/WSN/33, B/Yamagata/73, A/KAIM- 1 , or VSV/lndiana in PBS containing BSA at room temperature. After removal of the inoculum, cells were overlaid with medium (DMEM with 2OmM Hepes, 0.01 % DEAE Dextran, 0.001 % NaHC03) containing 1 % Oxoidagar and candidate peptides or small molecule compounds at the indicated concentrations.
  • medium DMEM with 2OmM Hepes, 0.01 % DEAE Dextran, 0.001 % NaHC03
  • Enzyme-Linked Immunosorbent Assay For the ELISA microwell plates (Pierce) were incubated with saturating concentrations of peptides at room temperature, washed and subsequently incubated at room temperature with HA-tagged PA. To obtain PA-HA, 293T cells were seeded into 94mm-dishes, transfected with the respective plasmid and treated with lysis buffer 24h post transfection as described in detail by Mayer et al. (2007).
  • HA-specific primary antibody Covance
  • a peroxidase- coupled secondary antibody Jackson l mmuno Research, Newmarket, UK
  • the competition ELISA was carried out as described above with the exception that the candidate peptide or small molecule competitor compound were added to wells of the plate with bound peptides prior to addition of the cell extract containing HA-tagged PA subunits.
  • the test sample includes a known binding pair of proteins or protein subunits including a fluorescent label, which can be analyzed according to a preferred embodiment of the present invention by fluorescence polarization.
  • a fluorescent label which can be analyzed according to a preferred embodiment of the present invention by fluorescence polarization.
  • PBl Influenza A virus polymerase subunit
  • PA subunit PA
  • the test sample is then contacted with a candidate peptide or small molecule inhibitor com pound and the resulting fluorescence polarization is determined.
  • the ability of the compound to cause dissociation of or otherwise interfere with or prevent binding of the proteins or protein subunits is monitored by fluorescence polarization (FP).
  • FP measurements allow for discrimination between fluorescently labeled bound and unbound proteins, peptides, subunits or fragments thereof.
  • the FP of the fluorescently labeled first fragment rotates rapidly in solution and, therefore, has randomized photo-selected distributions, which result in the small observed FP.
  • the rotation of the fluorescently labeled first fragment slows and the fluorescence polarization increases. Accordingly, disruption of the subunit interaction by a test compound provides a decrease in the fluorescence polarization, which is indicative of inhibition of the protein interactions.
  • the FP measurements in the presence of a test compound can be compared with the FP measurements in the absence of the test compound. Comparison can be made manually by the operator or automatically by a computer, especially in high throughput assays using 384-well plates.
  • PA protein purification influenza
  • a virus polymerase subunit PA was cloned into a suitable expression vector with a C-terminally attached 6xHis-linker or hemagglutinine epitope (HA).
  • Human 293T cells were transfected with the plasmid.
  • Cell lysates were prepared 24 hours post transfection using lysis buffer (2OmM TrisHCI pH 7.5, 1 0OmM NaCI, 0.5mM EDTA, 0.5% NP-40, I mM DTT and 1 % Protase inhibitor mix)
  • PA subunit was bound to Ni- or anti-HA-agarose and washed with lysis buffer without protease mix.
  • PA-protein was concentrated when necessary using Vivaspin20 5OK columns and frozen at -8O 0 C until further use. After thawing, the elution buffer was exchanged to low fluorescent grade reagents and any HA-peptide was removed simultaneously using 1 0-DC Bio-Gel columns.
  • Fluorescently labeled peptide corresponding to the 25 first N-terminal amino acids of Influenza A virus polymerase subunit PBl at 3 nM concentration was added to l O ⁇ M HA-PA in 2OmM TrisHCI pH 7.5, 1 5OmM NaCI, 0.5mM EDTA, I mM DTT, 5% Glycerol and l OOmg/ml bovine gamma globulin. The mix was distributed into black 384-well plates to a total volume of 20 ⁇ l per well and kept on ice. Test compounds solved in DMSO were added to a final concentration of 25 ⁇ M. After incubation for 1 0 minutes at room temperature, plates were read using an Infinite F200 reader (Tecan). FP values of the wells containing test compounds were compared to wells without test compounds, without DMSO and with peptide only.
  • Sequence alignment Alignments were performed with MUSCLE as described in Edgar (2004) using the full-length sequences provided from the public influenza virus database (https://www.ncbi.nlm.nih.gov/genomes/FLU/FLU.html).
  • H Ep-2 cells obtained from ATCC were seeded in 96-well plates (1 .5 x 1 04 cells per well) and grown in MEM-alpha medium containing 1 0% FBS (Cibco-BRL) for 24 h.
  • MEM-alpha medium containing 1 0% FBS (Cibco-BRL) for 24 h.
  • 500 pfu of RSV Long strain obtained from ATCC were added in 50 ⁇ l of OptiMEM (Cibco-BRL) for 1 h. Cells were then incubated in the presence of a serial dilution of compounds (from 1 00 to 0.14 ⁇ M) in MEM-alpha containing 2% FBS for 72 h.
  • Protein-protein interactions are crucial to most, if not all, biological processes. Of the roughly 30,000 protein sequences that comprise the human proteome, only about 1 % have been successfully targeted with small-molecule drugs. Yet, most of the conventional targets in drug discovery fall into the same few structural or functional families such as enzymes or C protein-coupled receptors (CPCRs). They typically share the property that the natural substrates or ligands, with which they interact are themselves small organic molecules. Historically there has been notably little success in developing drug-like inhibitors of proteins whose natural ligands are other proteins.
  • the present invention uses the fact that proteomes of many viruses and PPIs crucial for viral replication are described in the literature. For any proteome of interest, this data is according to the novel method supplemented with proteomic approaches for identification of PPIs like yeast two-hybrid or co-immuno precipitation screening in order to identify potential target regions for development of PPI inhibitors. Subsequently, a unique combination of phylogenetic analysis and structure prediction or structure analysis (where applicable) of the protein partners involved detects druggable protein-binding domains.
  • druggable denotes preferably protein-binding domains which can be blocked, altered or modified by small molecules in a way that the protein-protein interaction is inhibited or disrupted.
  • small molecules denotes organic molecules, preferably synthetic organic molecules (not peptides), which have a molecular weight below 1 500, preferably below 1 000 and most preferred below 500 u. It has been found, that these domains bear a couple of characteristic features: (i) helical structure, (ii) hydrophobic character and (iii) high conservation among all virus strains. It has been shown that they tend to be located at a term inal end of the protein or are located on their surface. The peptides corresponding to these potential binding domains are synthesized in an overlapping way and tested for their ability to bind the protein partner involved in the PPI.
  • peptides resembling short, (less than 20 amino acids) continuous binding domains are identified, these are used for the development of a binding assay, preferably an ELISA or fluorescence polarization (FP) assay, which is afterwards employed in a high-throughput screening campaign for small molecule and/or peptidic inhibitors of the PPI.
  • a binding assay preferably an ELISA or fluorescence polarization (FP) assay
  • PPI inhibitors identified by the novel method according to the present invention could offer a particular advantage when it comes to antivirals since it should be safe to assume that resistance development occurs at a much slower pace.
  • the ELISA assay described above for the influenza peptides was repeated with a number of small molecule com pounds obtained from corresponding compound libraries from Maybridge Ltd., Cambridge, UK (www.maybridge.com).
  • the tested compounds are listed in Ta ble 4, together with their systematic name, the product code, and the result for the ELISA assay.
  • PKE091 4-(3,4-d ⁇ fluorophenyl)-2-[5-(2-th ⁇ enyl)-2-th ⁇ enyl]-l ,3-th ⁇ azole SEW04837 no
  • PKE092 4-(tert-butyl)-2-[5-(2-th ⁇ enyl)-2-th ⁇ enyl]-l ,3-th ⁇ azole SEW04838 yes
  • PKEl 44 4-methyl-5-(l H-pyrazol-5-yl)-2-(2-th ⁇ enyl)-l ,3-th ⁇ azole SPB0261 2 no
  • PKE 064 and PKE 296 are identical. However, PKE 064 was obtained from a commercial compound library with restricted degree of purity and a considerable shelf time, while PKE 296 is the freshly synthesized, pure compound (>99%). It is assumed that the difference in the activities of PKE 064 and PKE 296 are due to impurities in PKE 064.
  • the assessed class of compounds seems to be effective in the inhibition of replication of certain virus types. A number of compounds very effectively inhibited the replication of influenza virus, particularly influenza A. Surprisingly it was found that a number of other compounds are also effective in the inhibition of RSV replication, and certain compounds inhibit the replication of both virus types.
  • Ar 1 is either a thiophene, a substituted thiophene, a furan or a substituted furan.
  • M can be a boronic acid, a boronic ester, or a stannyl functional group.
  • RI can be H, CH 3 , te/t-butyl, or CF 3 group.
  • Ester compounds (I) Compounds of formula (I), in the ester series, are generally prepared according to
  • a classic palladium (Pd) catalyzed coupling reaction such as a Suzuki coupling reaction or a Stille coupling reaction leads to bicyclic ester b.
  • Pd catalyst like tetrakis palladium triphenylphosphine
  • a base like aqueous NaHCO 3 or cesium fluoride
  • a classic organic solvent such as N 1 N- dimethylformamide or 1 ,2-Dimethoxyethane.
  • Brominated compound c is achieved by direct bromina- tion, using Br 2 in acetic acid media, /V-bromosuccinimide in a usual solvent, or copper (II) bromide in the presence of a nitrite in acetonitrile. Then another classic Pd catalyzed coupling reaction, a Suzuki reaction or a Stille reaction, yields the expected formula (I) compounds.
  • bicyclic systems Ar 2 -Ar I -M are directly available, as for thienyl-thienyl-M.
  • compounds of formula (I) are prepared from compound a by a one pot Suzuki or Stille coupling reaction, using classic Pd catalyst, such as tetrakis palladium triphenylphosphine, in the presence of a base like NaHCOs or cesium fluoride, in a usual solvent like /V,/V-dimethylformamide or 1 ,2-dimethoxyethane.
  • Acid compounds (II) Hydrolysis of the ester functional group of compounds (I) to the corresponding carboxylic acid (II) is made with lithium hydroxide in a methanol/water solution or in a tetra hyd rofu- ran/water solution. All acid compounds can be prepared as sodium salts or ammonium salts by usual procedures.
  • Alcohol compounds (III) Reduction of the ester functional group of compounds (I) to the corresponding alcohol is achieved in the presence of lithium aluminum hydride in a classic solvent such as tetrahy- drofuran, giving the compounds of formula (III).
  • Amide compounds (IV) The ester function of the brominated compound c is hydrolyzed with lithium hydroxyde to the corresponding carboxylic acid d. Then, the formation of the amid compound e can be achieved by classical reactions, such as formation of an acid chloride followed by addition of an amine, in the presence of a strong base like triethylamine, in a classical solvent like acetone or dichloro- methane. A Pd catalyzed coupling reaction like a Suzuki reaction or a Stille reaction gives the final compounds of formula (IV).
  • acetyl compounds of formula (V) follows the reaction scheme shown above.
  • the 2- aminothiazole f is brominated via a Sandmeyer reaction (EP 1 894930, 2008, or Organic Letters, 2003, 7963, or J. Chem. Soc Perkin Trans. J , 1 982, 1 59), using copper (II) bromide or NaBr/CuS0 4 , in the presence of a nitrite, either isoamylnitrite or sodium nitrite, in a solvent like acetonitrile.
  • the same synthesis steps as for formula (I) compounds apply: Pd catalyzed coupling reaction, bromination, and final Pd catalyzed coupling reaction lead to com pounds (V).
  • ester compounds (I) and acid compounds (II) follow the reaction scheme shown above.
  • M can be a boronic acid, a boronic ester, or a stannyl functional group.
  • Rl can be H, CH 3 , tert-butyl, or CF 3 group.
  • Starting 2-bromothiazole ester a undergoes a Pd catalyzed coupling reaction, like a Suzuki or a Stille reaction, to yield the alcohol j.
  • This alcohol is activated by use of triflic anhydride in the presence of 2,6-lutidine and DMAP in a solvent like dichloromethane (J. Chem.
  • Ar 1 is a thiophene
  • X can be H or Br.
  • Amides of formula (Vl) are prepared according to classic reaction, like peptide coupling reaction (J. Am. Chem. Soc, 2000, 6382-6394) using HOBt and EDCI in a usual solvent such as /V,/V-dimethylformamide.
  • X Br
  • a Pd catalyzed coupling reaction like a Suzuki or a Stille reaction leads to compounds of formula (VII).
  • Methylation of the nitrogen is achieved by using sodium hydride and methyl iodide in /V,/V-dimethylformamide, affording compounds of formula (VIII).
  • a classic hydrolysis of the ester function leads to compounds of formula (IX).
  • Ar1 , Ar2 substituted (or not) thiophene, phenyl, pyridine, A r s R9 benzothiophene, Ar 2 - ⁇ 1 - ⁇ V"
  • R1 ,R2 H, Me, COOEt, COOH, N R1
  • ArI is a thiophene, and either Rl or R2 or both can be a H, CH3, ester, acid, ester isoster or acid isoster (tetrazole, isoxazole hydroxamic acid or derivative).
  • Rl or R2 or both can be a H, CH3, ester, acid, ester isoster or acid isoster (tetrazole, isoxazole hydroxamic acid or derivative).
  • the synthesis of compounds of formula (Vl) follows the reaction scheme shown above. 2-Aminothiazole o is brominated via a Sandmeyer reaction. Then, the same synthesis steps as for formula (I) compounds apply: Pd catalyzed coupling reaction, bromination, and final Pd catalyzed coupling reaction lead to com pounds (Vl).
  • R1 ,R2,R 4 H, Me, CF 3 , COOEt, COOH, CHO,
  • ArI is a thiophene and Ar2 is a thiophene or a benzo[b]thiophene
  • term inal 2-linked-l ,3-thiazole has been replaced by other substituted cycles, such as thiophene, 5-lin ked-1 ,3- thiazole, 1 ,3,4-thiadiazole, or substituted phenyl.
  • the scheme shown above summarizes these structures.
  • the syntheses of compounds (X), (Xl), (XII) and (XIII) follow the same synthetic pathway described in reaction scheme 2 for thiazole derivatives.
  • Example Al -1 -1 l -(2-bromo-4-methyl-l ,3-thiazol-5-yl)-ethanone: To a mixture of 5-acetyl-2-amino-4- methyl-thiazole (4.50 g, 28.9 mmol) and anhydrous copper (II) bromide (6.46 g, 28.9 mmol) in dry ace- tonitrile (1 20 ml) was added isoamyl nitrite (1 1 .6 ml, 86.3 mmol). The mixture was stirred at 60 0 C for 4 hours and at room temperature for 1 6 hours.
  • Example Al -1 -2 / PKE 143 l -(4-methyl-2-thiophen-2-yl-thiazol-5-yl)-ethanone: To a mixture of thio- phene-2-boronic acid (3.37 g, 26.3 mmol), l -(2-bromo4-methyl-thiazol-5-yl)-ethanone Al -1 -1 (4.14 g, 1 8.8 mmol), and 2M aqueous Na 2 CO 3 (20 ml, 40.0 mmol ) in 1 30 ml of a DME-H 2 O solution (9/1 ) degassed under argon, was added Pd(PPh 3 J 4 (1 .09 g, 0.94 mmol).
  • reaction mixture was degassed under argon and heated up to 90 0 C for 1 6 h.
  • the reaction mixture was quenched by addition of water, diluted with EtOAc, filtered over celite and extracted with EtOAc. The organic layer was washed twice with water, dried over MgSO 4 and concentrated in vacuo. Chromatography of the residue on silica gel (cyclohexane-EtOAc: 8/2) afforded a yellow solid (83%).
  • Example Al -1 -3 l -[2-(5-Bromo-thiophen-2-yl)-4-methyl-l ,3-thiazol-5-yl]-ethanone: To a mixture of l -(4- methyl-2-thiophen-2-yl-thiazol-5-yl)-ethanone Al -1 -2 (3.25 g, 14.5 mmol) and anhydrous copper (II) bromide (3.25 g, 14.5 mmol) in dry acetonitrile (1 00 ml) was added isoamyl nitrite (1 .70 ml, 1 2.6 mmol). The mixture was stirred at 60 0 C for 20 hours. The solvent was evaporated in vacuo and the residue was purified by silica gel chromatography (EtOAc-cyclohexane: 2/8) to afford a yellow solid
  • Example Al -2 / PKE 296 l -(2-[2,2']Bithiophen-5-yl4-methyl-thiazol-5-yl)-ethanone: To a mixture of thiophene-2-boronic acid (0.1 2 g, 0.92 mmol), l -[2-(5-Bromo-thiophen-2-yl)-4-methyl-thiazol-5-yl]- ethanone Al -1 -3 (0.20 g, 0.66 mmol), and 2M aqueous Na 2 CO 3 (0.66 ml, 1 .22 mmol) in 1 0 ml of a DME-H 2 O (9/1 ) solution degassed under argon was added Pd(PPh 3 J 4 (38 mg, 33 mol).
  • reaction mixture was degassed under argon and heated up to reflux for 1 8 h.
  • the reaction mixture was quenched by addition of water, diluted with EtOAc, filtered over celite and extracted with EtOAc. The organic layer was washed twice with water, dried over MgSO 4 and concentrated in vacuo. Chromatography of the residue on silica gel (cyclohexane-EtOAc: 80/20) afforded a yellow solid (83%).
  • Example Al -3 / PKE 321 l -[4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazol-5-yl]-ethanone: Method as for Al -2, using pyridine-3-boronic acid; yellow solid (56%).
  • Example Al -4 / PKE 329 l - ⁇ 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazol-5-yl]-ethanone: Method as for Al -2, using 4-methoxy-phenyl boronic acid; yellow solid (35%).
  • Example Al -5 / PKE 346 l -[2-(5'-acetyl-[2,2']bithiophenyl-5-yl)-4-methyl-thiazol-5-yl]-ethanone: Method as for Al -2, using 2-acetyl-thiophene-5-boronic acid; yellow solid (22%).
  • Mp 1 64°C-1 66°C.
  • Example Al -6 / PKE 352 l - ⁇ 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazol-5-yl ⁇ - ethanone: Method as for Al -2, using 2-trifluoromethoxy-phenyl boronic acid; yellow solid (91 %).
  • Mp 1 01 0 C-1 03 0 C.
  • Example Al -7 / PKE 350 l -[2-(5-Benzo[b]thiophen-2-yl)-4-methyl-thiazol-5-yl]-ethanone: Method as for Al -2, using benzo[b]thiophene-2-boronic acid; yellow solid (66%).
  • 1 H NMR (DMSO) ⁇ : 8.00-7.97 (m, 1 H), 7.88-7.84 (m, 3H), 7.54 (d, J 4.0 Hz, 1 H), 7.43-7.39 (m, 2H), 2.68 (s, 3H), 2.56 (s, 3H).
  • Example Al -8 / PKE 385 l -[4-Methyl-2-(5'-methyl-[2,2']bithiophenyl-5-yl)-thiazol-5-yl]-ethanone: Method as for Al -2, using 5-methyl-thiophene-2-boronic acid; yellow solid (14%).
  • Mp 1 27 °C-1 30 0 C.
  • Example Al -9 / PKE 329 l -[4-Methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazol-5-yl]-ethanone hydrochloride salt (hydrochloride of compound PKE 321 ): Method as for Bl -60, using l -[4-methyl-2-(5-pyridin-3-yl- thiophen-2-yl)-thiazol-5-yl]-ethanone Al -3 / PKE 321 ; yellow solid (89%).
  • Example A2-1 -1 2-Bromo-4-t ⁇ t-butyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -1 , using 2-amino-4-tert-butyl-thiazole-5-carboxylic acid ethyl ester (prepared according to Journal of the Chemical Society, Perkin Trans I, 1 982, 1 52-1 64); white solid (94%).
  • MS (ESI + ) m/z: 291 .9-293.5 [M + H] + .
  • Example A2-1 -2 4-t ⁇ t-Butyl-2-thiophen-2-yl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al - 1 -2, using 2-bromo4-tert-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -1 ; beige solid (96%).
  • Example A2-1 -3 2-(5-Bromo-thiophen-2-yl)-4-tert-butyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -2, using 4-tert-butyl-2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester A2-1 -2; yellow solid (98%).
  • MS (ESI + ) m/z: 373.9-375.9 [M + H] + .
  • Example A2-M / PKE 259 2-[2,2 ']Bithiophenyl-5-yl-4-t ⁇ t-butyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-thiophen-2-yl)4-te/t-butyl-thiazole-5-carboxylic acid ethyl ester A2- 1 -3; yellow solid (75%).
  • Example A2-2 / PKE 357 2-[2,2 ']Bithiophenyl-5-yl-4-tert-butyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[2,2 ']bithiophenyl-5-yl-4-t ⁇ t-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -4; yellow solid (82%).
  • Example A2-3 / PKE 293 4-te/t-Butyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzene boronic acid and 2-(5-bromo-thiophen-2-yl)4-tert-butyl-thiazole-5- carboxylic acid ethyl ester A2-1 -3; yellow solid (91 %).
  • Example KlA / PKE 367 4-tert-Butyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-te/t-butyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A2-3; pale yellow solid (77%).
  • 1 H NMR (DMSO) ⁇ : 1 3.40 (br s, 1 H), 7.76-7.73 (m, 3H), 7.59 (d, J 3.9 Hz, 1 H), 7.46-7.35 (m, 3H), 1 .46 (s, 9H).
  • Example A2-5 / PKE 316 4-ferf-Butyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using pyridine-3-boronic acid and 2-(5-bromo-thiophen-2-yl)4-t ⁇ t-butyl- thiazole-5-carboxylic acid ethyl ester A2-l -3; yellow solid (66%).
  • Example A2-6 / PKE 320 4-te ⁇ f-Butyl-2-[5-(4-cyano-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-cyanobenzene boronic acid and 2-(5-bromo-thiophen-2-yl)-4-tert- butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (66%).
  • Example A2-7 / PKE 331 4-te ⁇ f-Butyl-2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylic acid ethyl ester: Method as for Al -2, using 4-trifluoromethoxy-phenyl boronic acid and 2-(5- bromo-thiophen-2-yl)4-tert " -butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (92%).
  • Example A2-8 / PKE 369 4-te ⁇ f-Butyl-2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-te ⁇ f-butyl-2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]- thiazole-5-carboxylic acid ethyl ester A2-7; yellow solid (27%).
  • Example A2-9 / PKE 332 4-te ⁇ f-Butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 3-methoxy-phenyl boronic acid and 2-(5-bromo-thiophen-2-yl)-4- te/t-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (60%).
  • Example A2-1 0 / PKE 344 4-te ⁇ f-Butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-te ⁇ f-Butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester A2-9; yellow solid (95%).
  • Example A2-1 1 / PKE 354 4-te ⁇ Butyl-2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methoxy-phenyl boronic acid and 2-(5-bromo-thiophen-2-yl)-4- te/t-butyl-thiazole-S-carboxylic acid ethyl ester A2-1 -3; green solid (79%).
  • Example A2-1 2 / PKE 355 4-te ⁇ f-Butyl-2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-te ⁇ f-Butyl-2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester A2-1 1 ; yellow solid (81 %).
  • Example A2-1 3 / PKE 358 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)4-t ⁇ t-butyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen- 2-yl)-4-t ⁇ t-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (83%).
  • Example A2-14 / PKE 379 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-t ⁇ t-butyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-tert-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 3; yellow solid (86%).
  • Example A2-1 5 / PKE 362 4-te ⁇ f-Butyl-2-(5 '-methyl-[2,2 ' ]bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 5-methylthiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-4- te/t-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (24%).
  • Example A2-1 6 / PKE 365 4-te/t-Butyl-2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]-l ,3-thiazole-5- carboxylic acid ethyl ester
  • Example A2-1 7 / PKE 356 4-te ⁇ f-Butyl-2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-te/t-Butyl-2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]- i o thiazole-5-carboxylic acid ethyl ester A2-1 6; yellow solid (75%).
  • Example A2-1 8 / PKE 370 2-(5 '-Acetyl-[2,2 ']bithiophenyl-5-yl)-4-te ⁇ f-butyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-acetyl-thiophene-5-boronic acid and 2-(5-bromo-thiophen-2-yl)-4- 15 te/t-butyl-thiazole-S-carboxylic acid ethyl ester A2-1 -3; yellow solid (27%).
  • Example A2-1 9 / PKE 377 4-te ⁇ f-butyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 3-quinoline boronic acid and 2-(5-bromo-thiophene-2-yl)-4-tert-butyl- 20 thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (27%).
  • Example A2-20 / PKE 380 4-te ⁇ f-butyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid: 25 Method as for B2-1 , using 4-te ⁇ f-butyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A2-1 9; yellow solid (92%).
  • Example A2-21 / PKE 345 Sodium 4-te ⁇ butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylate (sodium salt of compound PKE 344): Method as for B2-34, using 4-t ⁇ t-butyl-2-[5-(3- methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid A2-1 0 / PKE 344; yellow solid (75%).
  • Example A2-22 / PKE 31 9 4-te ⁇ f-Butyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester hydrochloride salt (hydrochloride of compound PKE 31 6): Method as for Bl -60, using 4-te/t-butyl- 2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A2-5 / PKE 31 6; yellow solid (60%).
  • Example A3-1 -1 2-Bromo-4-trifluoromethyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al - 1 -1 , using ethyl 2-amino4-trifluoromethyl-thiazole-5-carboxylate; white solid (71 %).
  • Example A3-1 -2 / PKE 470 2-[2,2 ']Bithiophenyl-5-yl-4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2,2'-bithiophene-5-boronic acid and 2-bromo4-trifluoromethyl- thiazole-5-carboxylic acid ethyl ester A3-1 -1 ; green solid (57%).
  • Example A3-2 / PKE 472 2-[2,2 ']Bithiophenyl-5-yl4-trifluoromethyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[2,2 ']Bithiophenyl-5-yl4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-1 -2; yellow solid (27%).
  • Example A3-3-1 2-Thiophen-2-yl4-trifluoromethyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2-bromo4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-1 -1 ; beige solid (68%).
  • Example A3-3-2 2-(5-Bromo-thiophen-2-yl)-4-trifluoromethyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -I -2, using 2-thiophen-2-yl-4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-3- 1 ; beige solid (99%).
  • Example A3-3-3 / PKE 506 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-4-trifluoromethyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo- thiophen-2-yl)4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-3-2; yellow solid (34%).
  • Example A3-4 / PKE 505 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-4-trifluoromethyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-trifluoromethyl- thiazole-5-carboxylic acid ethyl ester A3-3-3; yellow solid (53%).
  • Example A4-1 / PKE 21 2 2-[2,2']bithiophenyl-5-yl-thiazole-5-ca rboxylic acid ethyl ester: Method as for Al -1 -2, using 2,2'-bithiophene-5-boronic acid and ethyl 2-bromothiazole-5-carboxylate; green solid (4%).
  • Example A4-2 / PKE 21 1 2-[2,2']bithiophenyl-5-yl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[2,2']bithiophenyl-5-yl-thiazole-5-carboxylic acid ethyl ester A4-1 ; orange solid (80%).
  • Example A4-3-1 2-thiophen-2-yl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using ethyl 2-bromo-l ,3-thiazole-5-carboxylate; yellow solid (66%).
  • Example A4-3-2 2-(5-bromo-thiophen-2-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -I - 2, using 2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester A4-3-1 ; yellow solid (96%).
  • MS (ESI + ) m/z: 31 7.9 - 31 9.9 [M + H] + .
  • Example A4-3-3 / PKE 314 2-(5-Phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzene boronic acid and 2-(5-bromo-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A4-3-2; yellow solid (25%).
  • Example A4-4 / PKE 31 5 2-(5-Phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A4-3-3; yellow solid (83%).
  • Example A4-5 / PKE 371 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)- thiazole-5-carboxylic acid ethyl ester A4-3-2; yellow solid (1 6%).
  • Example A4-6 / PKE 214 Sodium 2-[2,2 ' ]bithiophenyl-5-yl-thiazole-5-carboxylate (sodium salt of compound PKE 21 1 ): Method as for B2-34, using 2-[2,2']b ithiophenyl-5-yl-thiazole-5-carboxylic acid A4-2 / PKE 21 1 ; yellow solid (83%).
  • Example A5-1 -1 2-Bromo-l ,3-thiazole4-carboxylic acid ethyl ester: Method as for Al -1 -1 , using 2- amino-thiazole-4-carboxylic acid ethyl ester; white solid (55%).
  • Example A5-2 / PKE 456 2-[2,2']Bithiophenyl-5-yl-thiazole4-carboxylic acid: Method as for B2-1, using 2-[2,2']bithiophenyl-5-yl-thiazole-4-carboxylic acid ethyl ester A5-1-2; pale yellow solid (100%).
  • Example A5-3 / PKE 448 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole: Method as for Bl-M, using 2- bromo-4-methyl-l,3-thiazole and 2,2'-bithiophenyl-5-boronic acid; beige solid (30%).
  • Example A5-4-1 4-Methyl-2-thiophen-2-yl-l,3-thiazole: Method as for Bl-M, using 2-bromo4-methyl- 1,3-thiazole; colourless oil (57%).
  • Example A5-4-2 2-(5-Bromo-thiophen-2-yl)-4-methyl-l,3-thiazole: Method as for BM -2, using 4-methyl- 2-thiophen-2-yl-thiazole A5-4-1; beige solid (88%).
  • Example A5-4-3 / PKE 489 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazole: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole A5-4-2; yellow solid (67%).
  • Example A5-5 / PKE 462 2-[2,2']bithiophenyl-5-yl-5-methyl-thiazole: Method as for Bl-M, using 2- bromo-5-methyl-l ,3-thiazole and 2,2'-bithiophenyl-5-boronic acid; brown solid (36%).
  • Example A5-6-1 5-Methyl-2-thiophen-2-yl-l ,3-thiazole: Method as for Bl-M, using 2-bromo-5-methyl- 1 ,3-thiazole; colourless oil (74%).
  • Example A5-6-2 2-(5-Bromo-thiophen-2-yl)-5-methyl-l ,3-thiazole: Method as for Bl -1 -2, using 5-methyl- 2-thiophen-2-yl-thiazole A5-6-1 ; yellow solid (58%).
  • MS (ESI + ) m/z: 260.0-261 .9 [M + H] + .
  • Example A5-6-3 / PKE 488 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-5-methyl-thiazole: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-5-methyl-thiazole A5-6-2; yellow solid (57%).
  • Example A5-7-1 2-Bromo-5-methyl-l ,3-thiazole-4-carboxylic acid ethyl ester: Method as for Al -1 -1 , us- ing 2-amino-5-methyl-thiazole4-carboxylic acid ethyl ester; yellow solid (83%).
  • Example A5-7-2 / PKE 540 2-[2,2 ']Bithiophenyl-5-yl-5-methyl-l ,3-thiazole-4-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2-bromo-5-methyl-thiazole-4-carboxylic acid ethyl ester A5-7-1 and 2,2'- bithiophenyl-5-boronic acid; yellow solid (84%).
  • Example A5-8 / PKE 541 2-[2,2 ']Bithiophenyl-5-yl-5-methyl-l ,3-thiazole4-carboxylic acid: Method as for B2-1 , using 2-[2,2 ' ]bithiophenyl-5-yl-5-methyl-thiazole-4-carboxylic acid ethyl ester A5-7-2; yellow solid (78%).
  • Example A5-9 / PKE 466 2-[2,2']-Bithophenyl-5-yl-benzothiazole: Method as for Bl -1 -1 , using 2,2'- bithiophenyl-5-boronic acid and 2-bromo-benzothiazole; yellow solid (42%).
  • Example A5-1 0-1 2-Bromo-6-methoxy-l ,3-benzo[d]thiazole: Method as for Al -1 -1 , using 2-amino-6- methoxy-benzothiazole; brown solid (26%).
  • MS (ESI + ) m/z: 243.9 - 245.9 [M + H] + .
  • Example A5-1 0-2 / PKE 463 2-[2,2 ']Bithiophenyl-5-yl-6-methoxy-benzothiazole: Method as for Bl -1 -1 , using 2,2'-bithiophenyl-5-boronic acid and 2-bromo-6-methoxy-benzothiazole A5-1 0-1 ; green solid (25%).
  • 1 H NMR (CDCI 3 ) ⁇ : 7.92 (brs, IH), 7.51 (br s, 1 H), 7.31-7.28 (m, 3H), 7.18 (brs, IH), 7.09 (br s, 1 H), 7.06 (t, J 4.0 Hz, 1 H), 3.89 (s, 3H).
  • Example A5-11-1 2-Thiophen-2-yl-l,3-benzo[d]thiazole: Method as for Bl-1-1 using 2-bromo- benzothiazole; white solid (74%).
  • Example A5-11-2 2-(5-Bromo-thiophen-2-yl)-l,3-benzo[d]thiazole: Method as for Bl-I -2, using 2- thiophen-2-yl-benzothiazole A5-11-1; white solid (97%).
  • Example A5-11-3 / PKE 527 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-benzothiazole: Method as for Bl-2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-benzothiazole A5-11-2; yellow solid (32%).
  • Example A5-12 / PKE 528 2-(5-Naphthalen-2-yl-thiophen-2-yl)-benzothiazole: Method as for Bl-2, using naphthalene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-benzothiazole A5-11-2; yellow solid (47%).
  • Example A5-13-1 2-Bromo4-methyl-thiazole-5-carbonitrile: 2-Bromo4-methyl-thiazole-5-carboxylic acid amide B3-11-1 (120 mg, 0.54 mmol) was dissolved in phosphorus oxychloride (8 ml) and reaction mixture was heated at 80 0 C for 3 h. After hydrolysis of the mixture in water (200 ml), the organics were extracted twice with EtOAc. Organic layers were washed twice with water, dried and concentrated in vacuo to afford a pale yellow solid (91%). Mp: 44°C46°C.
  • Example Bl -1 -1 / PKE 253 4-methyl-2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester: To a mixture of thiophene-2-boronic acid (0.32 g, 2.5 mmol), 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester
  • Example Bl -1 -2 / PKE 291 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: A mixture of 4-methyl-2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester Bl -1 -1 (0.8 g, 3.1 6 mmol) and bromine (0.55 g, 3.48 mmol) in 30 ml of glacial acetic acid was heated at 80 0 C for two hours. The 25 reaction mixture was poured into 200 ml of water and the precipitate formed was filtered off and dried to give a pale green solid (94 %).
  • Example Bl -1 -3 / PKE 290 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid: To a solution of 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (50 mg, 0.1 5 mmol) dissolved in a 8/2 mixture of methanol-water (25 ml) was added lithium hydroxide monohydrate (25 mg, 0.60 mmol). The reaction mixture was heated at reflux for 2 hours. The solvent was evaporated in vacuo, water (25 ml) was added, and the solution extracted with CH 2 CI 2 (3 x 1 5 ml).
  • Example Bl -2 / PKE 21 2 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: To a mixture, degassed under argon, of thiophene-2-boronic acid (0.1 1 g, 0.9 mmol), 2-(5-bromo-thiophen-2- yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (0.1 5 g, 0.5 mmol), and powdered CsF (0.28 g, 1 .9 mmol) in 1 0 ml of DME and 3 ml of water was added Pd(PPh 3 J 4 (37 mg, 0.03 mmol).
  • thiophene-2-boronic acid 0.1 1 g, 0.9 mmol
  • reaction mixture was degassed under argon and heated up to reflux for 1 8 h.
  • the reaction mixture was then diluted with EtOAc and a saturated aqueous solution of NaHCO 3 .
  • the organic layer was washed with brine, dried over MgSO 4 and concentrated under vacuum. Chromatography of the residue on silica gel (cyclohexane-EtOAc: 80/20) afforded a yellow solid (72%).
  • Example Bl -3 / PKE 254 2-[2,3']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using thiophene-3-boronic acid; yellow solid (7%).
  • Example Bl -4 / PKE 526 2-(5-furan-3-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using furane-3-boronic acid; yellow powder (99%).
  • Example Bl -5 / PKE 529 4-methyl-2-[2,2',5',2"]terthiophen-5-yl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using [2,2',5']tert-thiophene-2-boronic acid; yellow solid (69%).
  • Example Bl -7 / PKE 289 2-[5-(2-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-methoxyphenyl boronic acid; yellow powder (49%).
  • Example Bl -8 / PKE 288 2-[5-(2-cyano-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-cyanophenyl boronic acid; yellow solid (14%).
  • Example Bl -9 / PKE 287 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methoxyphenyl boronic acid; yellow solid (74%).
  • Example Bl -1 0 / PKE 301 2-[5-(2-fluoro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-fluorophenyl boronic acid; yellow powder (57%).
  • Example Bl -I l / PKE 297 4-methyl-2-(5'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 5-methyl-thiophene-2-boronic acid; yellow solid (72%).
  • Example Bl -1 2 / PKE 302 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid; yellow solid (43%).
  • Example Bl -1 3 / PKE 303 2-[5-(3-fluoro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-fluorophenyl boronic acid; yellow solid (67%).
  • Example Bl -14 / PKE 304 2-(5-benzofuran-2-yl-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzofurane-2-boronic acid; yellow powder (31 %).
  • Example Bl -1 5 / PKE 300 2-[5-(4-fluoro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-fluorophenyl boronic acid; yellow powder (5%).
  • Example Bl -1 6 / PKE 299 4-methyl-2-(5'-acetyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-acetyl-thiophene-5-boronic acid; yellow solid (44%).
  • Example Bl -1 7 / PKE 31 7 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 2-trifluoromethoxy-benzene boronic acid; yellow solid (62%).
  • Example Bl -1 8 / PKE 31 8 2-[5-(3-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 3-trifluoromethoxy-benzene boronic acid; yellow powder (30%).
  • Example Bl -1 9 / PKE 322 2-[5-(4-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-trifluoromethyl benzene boronic acid ; yellow powder (33%).
  • Example Bl -20 / PKE 324 2-(5-benzo[ l ,3]dioxol-5-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3,4-methylenedioxy benzene boronic acid; yellow solid (44%).
  • Example Bl -21 / PKE 325 2-[5-(2,3-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2,3-dimethoxyphenyl boronic acid; yellow solid (34%).
  • Example Bl -22 / PKE 326 2-[5-(2-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-trifluoromethyl-benzene boronic acid; yellow solid (42%).
  • Example Bl -23 / PKE 327 2-[5-(3-acetyl-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-acetylphenyl boronic acid; yellow powder (1 3%).
  • Example Bl -24 / PKE 333 4-methyl-2-(5-pyrimidin-5-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using pyrimidine-5-boronic acid; yellow powder (25%).
  • Example Bl -26 / PKE 349 2-[5-(3-formyl-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-formylphenyl boronic acid; yellow powder (1 6%).
  • Example Bl -27 / PKE 351 2-[5-(2,4-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2,4-dimethoxyphenyl boronic acid; yellow solid (64%).
  • Example Bl -28 / PKE 363 2-[5-(3-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-methoxyphenyl boronic acid; green solid (55%).
  • Example Bl -29 / PKE 378 4-methyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using quinoline-3-boronic acid; yellow powder (1 1 %).
  • Example Bl -30 / PKE 373 2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 4-trifluoromethoxy-benzene boronic acid; yellow powder (32%).
  • Example Bl -31 / PKE 372 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using naphthalene-2-boronic acid; yellow powder (26%).
  • Example Bl -32 / PKE 393 2-[5-(3-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-trifluoromethyl-benzene boronic acid; yellow powder (67%).
  • 1 H NMR (DMSO) ⁇ : 8.05 (s, 1 H), 8.03 (d, J 8.6 Hz, 1 H), 7.
  • Example Bl -33 / PKE 394 4-methyl-2-(4'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-thiophene-2-boronic acid; yellow powder (21 %).
  • Example Bl -34 / PKE 388 4-methyl-2-(5-m-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-methylbenzene boronic acid; yellow powder (46%).
  • Example Bl -35 / PKE 389 4-methyl-2-(5-naphthalen-l -yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using naphthalene-1 -boronic acid; yellow powder (57%).
  • Example Bl -36 / PKE 390 4-methyl-2-(5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methylbenzene boronic acid; yellow powder (77%).
  • Example Bl -37 / PKE 391 4-methyl-2-(5-o-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-methylbenzene boronic acid; yellow powder (48%).
  • Example Bl -38 / PKE 392 4-methyl-2-(3'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-methyl-thiophene-2-boronic acid; yellow powder (33%).
  • Example Bl -39 / PKE 398 4-methyl-2-(5'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-phenyl-thiophene-5-boronic acid; yellow powder (54%).
  • Example Bl -40 / PKE 401 2-[5-(2-chloro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-chloro-benzene boronic acid; yellow powder (24%).
  • Example Bl -41 / PKE 425 2-[5-(2,4-dimethyl-thiazol-5-yl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2,4-dimethyl-5-(4,4,5,5-tetramethyl-l ,3,2-dioxoborolan-2-yl)- 1 ,3-thiazole; yellow powder (46%).
  • Example B142 / PKE 427 4-methyl-2-[5-(4-methyl-2-phenyl-thiazol-5-yl)-thiophen-2-yl]-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-5-(4,4,5,5-tetramethyl-l ,3,2-dioxoborolan- 2-yl)-2-phenyl-l ,3-thiazole; yellow powder (86%).
  • Example Bl -43 / PKE 428 4-methyl-2-(4'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-phenyl-thiophene-5-boronic acid; yellow powder (65%).
  • Example Bl -44 / PKE 429 2-[5-(5-Ethoxycarbonyl4-methyl-thiazol-2-yl)-thiophen-2-yl]-indole-l - carboxylic acid tert-butyl ester: Method as for Bl -2, using /V-Boc-indole-2-boronic acid; yellow oil (35%).
  • Example Bl -45 / PKE 439 4-methyl-2-(5-thiazol-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: To a mixture of 2-(tributyl-stannyl)-l ,3-thiazole (0.25 g, 0.7 mmol) and 2-(5-bromo-thiophen-2-yl)-4- methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (0.1 7 g, 0.5 mmol) in 1 0 ml of DME degassed under argon was added Pd(PPh 3 J 4 (22 mg, 0.02 mmol).
  • reaction mixture was degassed under argon and heated up to reflux for 1 8 h.
  • the reaction mixture was then diluted with methylene chloride and water and the organic layer was dried and concentrated under vacuum. Chromatography of the residue on preparative silica TLC (cyclohexane-EtOAc: 80/20) afforded a yellow powder (42%).
  • Example Bl -46 / PKE 461 2-(5-benzo[b]thiophen-5-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-5-boronic acid; yellow solid (41 %).
  • Example Bl Al / PKE 493 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 5-methoxy-benzo[b]thiophene-2-boronic acid; yellow solid (56%).
  • Example Bl -48 / PKE 494 2-[5-(5-fluoro-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 5-fluoro-benzo[b]thiophene-2-boronic acid; yellow solid (31 %).
  • Example Bl -49 / PKE 501 2-[5-(6-hydroxy-naphthalen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 6-hydroxy-naphthalene-2-boronic acid; yellow solid (55%).
  • Example Bl -52 / PKE 258 4-Methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl es- ter: Method as for Bl -2, using pyridine-3-boronic acid; yellow solid (81 %).
  • Example Bl -53 / PKE 295 4-Methyl-2-(5-pyridin-4-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl es- ter: Method as for Bl -2, using pyridine-4-boronic acid; yellow solid (21 %).
  • Example Bl -54/ PKE 260 4-Methyl-2-[5-(l H-pyrazol4-yl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 1 W-pyrazole-4-boronic acid; yellow solid (14%).
  • Example Bl -55 / PKE 330 4-Methyl-2-[5-( 1 -methyl-1 H-pyrazol4-yl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using l -Methyl4-(4,4,5,5-tetramethyl-l ,3,2-dioxaborolan-2-yl)-l H- pyrazole; yellow solid (23%).
  • Example Bl -56 / PKE 402 2-[5-(3-Chloro-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-chlorobenzene boronic acid; yellow solid (26%).
  • Example Bl -57 / PKE 403 2-(5-Benzo[b]thiophen-3-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using l -benzothiophen-3-ylboronic acid; yellow solid (28%).
  • Example Bl -58 / PKE 405 4-Methyl-2-[5-(5-methyl-benzo[b]thiophen-2-yl)-thiophen-2-yl]-thiazole-5- ca rboxylic acid ethyl ester: Method as for Bl -2, using 5-methylbenzo[b]thiophen-2-boronic acid; yellow solid (31 %).
  • Example Bl -59 / PKE 406 2-[5-(4-Cyano-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-cyanobenzene boronic acid; yellow solid (30%).
  • Example Bl -60 / PKE 261 4-Methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester hydrochloride salt (hydrochloride of compound PKE 258): A 4M solution of hydrogen chloride in diox- ane (0.020 ml, 0.080 mmol) was added to a solution of 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole- 5-carboxylic acid ethyl ester Bl -52 / PKE 258 (25 mg, 0.076 mmol) in ethanol (2 ml) and dioxane (1 ml).
  • Example Bl -61 / PKE 294 4-Methyl-2-(5-pyridin-4-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester hydrochloride salt (hydrochloride of compound PKE 295): Method as for Bl -60, using 4-methyl-2-(5- pyridin-4-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -53 / PKE 295; yellow solid (68%).
  • Example B2-1 / PKE 193 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid: To a solution of 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -2 (1 20 mg, 0.36 mmol) dissolved in a 8/2 mixture of methanol-water (50 ml) was added lithium hydroxide monohydrate (50 mg, 1 .1 9 mmol). The reaction mixture was heated at reflux for 2 hours. The solvent was evaporated in vacuo, water (25 ml) was added, and the solution was washed with CH 2 Cb (3 x 1 5 ml).
  • Example B2-2 / PKE 262 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -52 / PKE 258; yellow needles (98%).
  • Example B2-3 / PKE 292 4-methyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -51 / PKE 257; yellow solid (44%).
  • Mp 236°C-238°C
  • ⁇ H NMR (DMSO) ⁇ : 1 3.29 (br s, 1 H), 7.68 (d, J 3.7 Hz, 1 H), 7.25 (br s, 2H), 6.82 (br s, 1 H), 2.59 (s, 3H), 2.45 (s, 3H).
  • Example B2-5 / PKE 31 3 2-(5'-acetyl-[2,2']bithiophen-5-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5'-acetyl-[2,2']bithiophen-5-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 6; ochre solid (95%).
  • Example B2-6 / PKE 361 : 2-[5-benzo[b]thiophen-2-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 2; orange solid (95%).
  • Example B2-7 / PKE 364 2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -28; yellow solid (72%).
  • Example B2-8 / PKE 366 2-[5-(2,4-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(2,4-dimethoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -27; yellow solid (97%).
  • Example B2-9 / PKE 368 2-(5-benzo[ l ,3]dioxol-5-yl-thiophen-2-yl)-4-methyl-thiazole-5-ca rboxylic acid: Method as for B2-1 , using 2-(5-benzo[ l ,3]dioxol-5-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -20; yellow solid (68%).
  • Example B2-1 0 / PKE 375 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -9; yellow solid (93%).
  • Example B2-1 1 / PKE 376 2-[5-(2-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(2-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -7; yellow solid (89%).
  • Example B2-1 2 / PKE 374 2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl -1 8; orange solid (86%).
  • Example B2-1 3 / PKE 347 2-[5-(3-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(3-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester Bl -32; yellow solid (94%).
  • Example B2-14 / PKE 382 4-methyl-2-(5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester BI -36; yellow solid (64%).
  • Example B2-1 5 / PKE 381 4-methyl-2-(3'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(3'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester Bl -38; orange solid (93%).
  • Example B2-1 6 / PKE 396 4-methyl-2-(5'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester Bl -39; ochre solid (50%).
  • Example B2-1 7 / PKE 397 4-methyl-2-(5-naphthalen-l -yl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-naphthalen-l -yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -35; pale yellow solid (50%).
  • Example B2-1 8 / PKE 404 2-(5-benzo[b]thiophen-3-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-3-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -57/ PKE 403; yellow solid (65%).
  • Example B2-1 9 / PKE 407 4-methyl-2-[5-(5methyl-benzo[b]thiophen-2-yl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-methyl-2-[5-(5methyl-benzo[b]thiophen-2-yl)-thiophen-2-yl]- thiazole-5-carboxylic acid ethyl ester Bl -58 / PKE 405; yellow solid (74%).
  • Example B2-20 / PKE 431 4-methyl-2-(4'-phenyl-[2,2']bithiophen-5-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4'-phenyl-[2,2']bithiophen-5-yl)-thiazole-5-carboxylic acid ethyl ester Bl 43; yellow solid (80%).
  • Example B2-21 / PKE 430 4-methyl-2-[5-(4-methyl-2-phenyl-thiazol-5-yl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-methyl-2-[5-(4-methyl-2-phenyl-thiazol-5-yl)-thiophen-2-yl]- thiazole-5-carboxylic acid ethyl ester Bl 42; yellow solid (75%).
  • Example B2-22 / PKE 440 2-[5-(l W-lndol-2-yl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(5-Ethoxycarbonyl4-methyl-thiazol-2-yl)-thiophen-2-yl]-indole-l -carboxylic acid te/t-butyl ester Bl 44; brown solid (47%).
  • Example B2-23 / PKE 449 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -31 ; pale yellow solid (81 %).
  • Example B2-24 / PKE 451 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl -1 7; yellow solid (1 00%).
  • Example B2-25 / PKE 537 2-(5-furan-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-furan-2-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -6; ochre 5 solid (82%).
  • Example B2-27 / PKE 468 2-(5-benzo[b]thiophen-5-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid: Method as for Bl -2, using 2-(5-benzo[b]thiophen-5-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid 15 ethyl ester Bl 46; brown solid (84%).
  • Example B2-28 / PKE 491 2-[5-(5-hydroxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: To a solution of 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-
  • Example B2-29 / PKE 492 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for Bl -2, using 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4- methyl-thiazole-5-carboxylic acid ethyl ester Bl 47; yellow solid (71 %).
  • Example B2-30 / PKE 495 2-[5-(5-fluoro-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for Bl -2, using 2-[5-(5-fluoro-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl 48; brown solid (72%).
  • Example B2-31 / PKE 507 2-[5-(6-hydroxy-naphthalen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for Bl -2, using 2-[5-(6-hydroxy-naphthalen-2-yl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl 49; yellow solid (72%).
  • Example B2-32 / PKE 524 2-(5-furan-3-ylthiophen-2-yl)4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-furan-3-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -4; yellow solid (88%).
  • Mp 214°C-21 7 °C.
  • Example B2-33 / 525 4-Methyl-2-[2,2',5',2"]tertiophen-5-yl-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-[2,2',5',2"]terthiophen-5-yl-thiazole-5-carboxylic acid ethyl ester Bl -5; orange solid (64%).
  • Example B2-34 / PKE 209 Sodium 2-[2,2 ' ]bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylate (sodium salt of compound PKE 1 93): To a suspension of 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid B2-1 / PKE 1 93 (430 mg, 1 .4 mmol) in water (20 ml) at room temperature is added sodium hy- droxide (56 mg, 1 .4 mmol). Reaction mixture is stirred at room temperature for 2 h. Solvents are evaporated to dryness to give a yellow solid (69%).
  • Example B2-35 / PKE 207 Ammonium 2-[2,2 ']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylate (ammonium salt of compound PKE 1 93): To a suspension of 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5- carboxylic acid B2-1 / PKE 193 in water (50ml) and ethanol (50 ml) at room temperature was added a 30% aqueous solution of ammonia, until dissolution of the precipitate. Solvents were evaporated to dryness to give a yellow solid (63%).
  • Example B2-36 / PKE 359 Sodium 4-methyl-2-(5 '-methyl-[2,2 ' ]bithiophenyl-5-yl)-thiazole-5-carboxylate (sodium salt of compound PKE 298): Method as for B2-34, using 4-methyl-2-(5'-methyl-[2,2']bithiophen- 5-yl]-thiazole-5-carboxylic acid B2 ⁇ / PKE 298; yellow solid (1 00%).
  • Example B2-37 / PKE 360 Sodium 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylate (sodium salt of compound PKE 262): Method as for B2-34, using 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)- thiazole-5-carboxylic acid B2-2 / PKE 262; yellow solid (1 00%).
  • Example B2-38 / PKE 348 Sodium 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-methyl-thiazole-5- carboxylate (sodium salt of compound PKE 361 ): Method as for B2-34, using 2-(5-benzo[b]thiophen-2- yl-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid B2-6 / PKE 361 ; yellow solid (1 00%).
  • Example B3-1 -1 / PKE 290 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid: To a solution of 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (50 mg, 0.1 5 mmol) dissolved in a 8/2 mixture of methanol-water (25 ml) was added lithium hydroxide monohydrate (25 mg, 0.60 mmol). The reaction mixture was heated at reflux for 2 hours. The solvent was evaporated in vacuo, water (25 ml) was added, and the solution extracted with CH 2 CI 2 (3 x 1 5 ml).
  • Example B3-1 -2 2-(5-bromo-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid (2-morpholin4-yl- ethyl)-amide: To a solution of 2-(5-bromo-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid B3-1 -1 (1 .64 mmol, 500 mg) in acetone (1 0 ml), between -1 0/-1 5 0 C, was added triethylamine (4.36 mmol, 0.6 ml), isobutylchloroformate (1 .89 mmol, 0.1 8 ml) followed by 2-aminoethyl morpholine (1 .64 mmol, 0.22 ml). The reaction mixture was stirred at room temperature. After completion of the reaction and concentration, the crude was purified by silica gel column chromatography (EtOAc-MeOH: 9/1 ) to
  • Example B3-1 -3 / PKE 1 89.1 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid (2-morpholin- 4-yl-ethyl)-amide: To a mixture, degassed under argon, of thiophene-2-boronic acid (0.1 6 g, 1 .3 mmol), 2-(5-bromo-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid (2-morpholin4-yl-ethyl)-amide B3-1 -2 (0.47 g, 1 .1 mmol), and powdered CsF (0.38 g, 2.5 mmol) in 1 0 ml of DME was added Pd(PPh 3 ) 4 (44 mg, 0.04 mmol, 3 mol percent).
  • Reaction mixture was degassed under argon and heated up to reflux for 1 8 h. Reaction mixture was then diluted with water and extracted with methylene chloride. Organic layer was dried and concentrated. Chromatography of the residue on silica gel (Et0Ac/Me0H/NH 4 0H: 90/9/1 ) afforded a yellow solid (1 5%).
  • Example B3-2 / PKE 500 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid (2-morpholin4-yl-ethyl)-amide: Method as for B3-1 -3, using benzo[b]thiophene-2-boronic acid; pale yellow solid (20%).
  • Example B3-3 / PKE 499 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid (2- morpholin-4-yl-ethyl)-amide: Method as for B3-1 -3, using naphthalene-2-boronic acid; yellow solid (59%).
  • Example B3-4-1 2-Bromo-4-methyl-l ,3-thiazole-5-carboxylic acid (3-morpholin4-yl-propyl)-amide: Method as for B3-1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid and 3-morpholin-4-yl propylamine; yellow oil (82%).
  • Example B3-4-2 / PKE 1 92.1 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid (3-morpholin- 4-yl-propyl)-amide: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and 2-bromo4-methyl- thiazole-5-carboxylic acid (3-morpholin4-yl-propyl)-amide B3-4-1 ; yellow solid (39%).
  • Example B3-5-1 2-Bromo4-methyl-l ,3-thiazole-5-carboxylic acid (3-imidazol-l -yl-propyl)-amide : Method as for B3-1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid and 3-imidazol-l -yl propylamine; color- less oil (61 %).
  • Example B3-5-2 / PKE 195 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid (3-imidazol-l -yl- propyl)-amide: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and 2-bromo4-methyl- thiazole-5-ca rboxylic acid (3-imidazol-l -yl-propyl)-amide B3-5-1 ; green solid (39%).
  • Example B3-6-1 2-Bromo-4-methyl-l ,3-thiazole-5-carboxylic acid ( 1 -ethyl-pyrrol id i n-2-yl methyl)-a m ide: Method as for B3-1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid and C-(I -ethyl-pyrrolidin-2-yl)- methylamine; pinkish oil (72%).
  • Example B3-6-2 / PKE 204 2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid (1 -ethyl- pyrrolidin-2-ylmethyl)-amide: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and 2-bromo-4- methyl-thiazole-5-carboxylic acid ( 1 -ethyl-pyrrol id i n-2-y lmethyl)-a m ide B3-6-1 ; yellow solid (1 2%).
  • Example B3-7-1 (2-Bromo4-methyl-l ,3-th i a zo l-5-y I )-[4-(4-f I uorophenyl)-piperazin-1 -yl]-methanone: Method as for B3-1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid and 1 -(4-fluorophenyl)- piperazine; white solid (1 5%).
  • 1 H NMR (CDCI 3 ) ⁇ : 6.95-6.88 (m, 2H), 6.87-6.81 (m, 2H), 3.71 (m, 4H),
  • Example B3-7-2 / PKE 208 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazol-5-yl-[4-(4-fluorophenyl)-piperazin- l -yl]-methanone: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and (2-bromo4-methyl- thiazole-5-yl)-[4-(4-fluoro-phenyl)-piperazin-l -yl]-methanone B3-7-1 ; yellow solid (3%). MS (ESI + ) m/z: 470.0 [M + H] + . Mp: 1 25°C-1 26°C.
  • Example B3-8 / PKE 21 5 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid isobutyl a mide: To a suspension of 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid B2-1 (307 mg, 1 .0 mmol) in dichloromethane (1 0ml) at room temperature are added oxalyl chloride (0.4 ml, 4.7 mmol) and N 1 N- dimethylformamide (0.4 ml). Reaction mixture is stirred at room temperature until complete dissolution (2h) and solvents are evaporated to dryness.
  • Residue is dissolved back in dichloromethane (1 0ml) and isobutylamine (0.6 ml, 6.0 mmol) is added. Reaction mixture is stirred at room temperature for 1 2 h, then diluted with EtOAc and washed twice with water. Organic layers are dried and evaporated. Solid obtained is washed with diethyl ether and dried to afford a yellow solid (33%).
  • Example B3-9-1 2-(5-bromo-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl amide: Method as for B3-1 -2, using ethylamine 2M solution in dioxane; white solid (55%).
  • Example B3-9-2 / PKE 535 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl amide: Method as for B3-1 -3, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2- yl)-4-methyl-thiazole-5-carboxylic acid ethyl amide B3-9-1 ; orange solid (72%).
  • Example B3-1 0-1 [2-(5-bromo-thiophen-2-yl)4-methyl-l ,3-thiazol-5-yl]-pyrrolidin-l -yl-methanone: Method as for B3-l -2, using pyrrolidine; yellow oil (51 %).
  • Example B3-1 0-2 / PKE 536 [2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazol-5-yl]-pyrrolidin- 1 -yl-methanone: Method as for B3-1 -3, using benzo[b]thiophene-2-boronic acid and [2-(5-bromo- thiophen-2-yl)4-methyl-thiazol-5-yl]-pyrrolidin-l -yl-methanone B3-1 0-1 ; yellow solid (28%).
  • Example B3-1 1 -1 2-bromo4-methyl-l ,3-thiazole-5-carboxylic acid amide: A suspension of 2-bromo-4- methyl-1 ,3-thiazole-5-carboxylic acid ethyl ester (0.60 g, 2.4 mmol) in a 30% aqueous ammonia solution (1 5 ml) was stirred at room temperature for 48 hours. Precipitate was filtered off, washed with water and dried to afford a white solid (94%).
  • Example B3-1 1 -2 / PKE 543 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid amide
  • Example B4-1 / PKE 213 (2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazol-5-yl)-methanol: To a suspension of lithium aluminium hydride (2.64 mmol, 1 00 mg) in dry TH F (1 0 ml) at 0 0 C, was added 2-
  • Example B4-2 / PKE 530 [2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-methyl-thiazole-5-yl] -methanol: Method as for B4-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 2; yellow solid (45%).
  • Example B5-1 / PKE 205 2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid (2-morpholin4-yl- ethyl)-ester: Method as for B3-8, using 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid B2-1 and 2-hydroxy-ethylmorpholine; yellow solid (1 9%).
  • Example Cl -1 -1 4-methyl-2-(3-methyl-thiophen-2-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -M , using 3-methyl-thiophene-2-boronic acid; pale yellow solid (65%).
  • Example Cl -1 -2 2-(5-bromo-3-methyl-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -I -2, using 4-methyl-2-(3-methyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Cl - 1 -1 ; white solid (99%).
  • Example Cl -1 -3 / PKE 414 4-methyl-2-(4-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-3-methyl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester CM -2 and thiophene-2-boronic acid; yellow solid (37%).
  • Example Cl -1 -4 / PKE 41 3 2-(5-Benzo[b]thiophen-2-yl-3-methyl-thiophen-2-yl)4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-3-methyl-thiophen-2-yl)4-methyl- thiazole-5-carboxylic acid ethyl ester Cl -1 -2 and benzo[b]thiophene-2-boronic acid; yellow solid (41 %).
  • Example CM -5 / PKE 424 4-Methyl-2-(3-methyl-5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-(5-bromo-3-methyl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester CM -2 and 4-methyl-benzene boronic acid; yellow solid (1 9%).
  • Example Cl -1 -6 / PKE 41 1 2-(5-Benzo[b]thiophen-2-yl-3-methyl-thiophen-2-yl)4-methyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-3-methyl-thiophen-2-yl)4-methyl- thiazole-5-carboxylic acid ethyl ester Cl -M; yellow solid (88%).
  • Example Cl -1 -7 / PKE 41 2 4-Methyl-2-(4-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester Cl -1 -3; yellow solid (90%).
  • Example Cl -2-1 4-Methyl-2-(4-methyl-thiophen-2-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 4-methyl-thiophene-2-boronic acid; beige solid (98%).
  • Example Cl -2-2 2-(5-Bromo-4-methyl-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -2, using 4-methyl-2-(4-methyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Cl - 2-1 ; beige solid (68%).
  • Example Cl -2-3 4-Methyl-2-(3-methyl-[2,2']bithiophenyl-5-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-4-methyl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Cl -2-2 and thiophene-2-boronic acid; yellow solid (28%).
  • Example Cl -2-4 / PKE 399 2-(5-Benzo[b]thiophen-2-yl4-methyl-thiophen-2-yl)4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo4-methyl-thiophen-2-yl)4-methyl- thiazole-5-carboxylic acid ethyl ester Cl -2-2 and benzo[b]thiophene-2-boronic acid; yellow solid (1 9%).
  • Example Cl -2-5 / PKE 395 4-Methyl-2-(3-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(3-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester Cl -2-3; ochre solid (45%).
  • Mp 224°C-227 °C dec.
  • Example Cl -3-1 / PKE 256 4-Methyl-2-thiophene-3-yl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester and thiophene-3-boronic acid; white solid (91 %).
  • Example Cl -3-2 2-(2-Bromo-thiophene-3-yl)4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: 4-Methyl- 2-thiophene-3-yl-thiazole-5-carboxylic acid ethyl ester Cl -3-1 (840 mg, 3.30 mmol) and bromine (53 mg, 3.30 mmol) in glacial acetic acid (40 ml) were heated at 70 0 C for 1 8 hours. Reaction mixture was concentrated. Residue was dissolved back in EtOAc (30 ml). Organic layers were washed twice with 1 0% aqueous K 2 CO 3 , 1 0% aqueous NaHSO 3 , and H 2 O. They were then concentrated, and dried under vacuum to give a pink solid (82 %).
  • Example Cl -3-3 / PKE 286 2-[2,2']-Bithiophenyl-3-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(2-bromo-thiophene-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Cl - 3-2 and thiophene-2-boronic acid; pink solid (30%).
  • Example Cl -3-4/ PKE 323 2-(5'-Acetyl-[2,2']-bithiophenyl-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(2-bromo-thiophene-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Cl -3-2 and 2-acetyl-thiophene-5-boronic acid; pale yellow solid (26%).
  • Example Cl -4 / PKE 426 4-Methyl-2-(4-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 4-phenylthiophene-2-boronic acid pinacol ester; pale yellow solid (23%).
  • Example C2-1 -1 4-Methyl-2-furan-2-yl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using furane-2-boronic acid; grey solid (65%).
  • Example C2-1 -2 2-(5-Bromo-furan-2-yl)4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -2, using 4-methyl-2-furan-2-yl-thiazole-5-carboxylic acid ethyl ester C2-1 -1 ; pink solid (65%).
  • Example C2-2 / PKE 523 2-(5-Benzo[b]thiophen-2-yl-furan-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-(5-bromo-furan-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C2-1 -2 and benzo[b]thiophene-2-boronic acid; pale yellow solid (1 5%).
  • Example C2-3 / PKE 533 2-(5-Benzo[b]thiophen-2-yl-furan-2-yl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-furan-2-yl)-4-methyl-thiazole-5-ca rboxylic acid ethyl ester C2-2; yellow solid (99%).
  • Example C3-1 / PKE 450 4,4'-Dimethyl-2'-phenyl-[2,5']bithiazolyl-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 4-methyl-2-phenyl-5- (4,4,5,5-tetramethyl-l ,3,2-dioxoborolan-2-yl)-l ,3-thiazole; pale yellow solid (54%).
  • Example C3-2 / PKE 447 4,4'-Dimethyl-2'-phenyl-[2,5']bithiazolyl-5-carboxylic acid: Method as for B2- 1 , using 4,4'-dimethyl-2'-phenyl-[2,5']bithiazolyl-5-carboxylic acid ethyl ester C3-1 ; white solid (75%).
  • Example C4-1 -1 2-(3-Hydroxy-phenyl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 3-hydroxyphenyl-boronic acid; white solid (49%).
  • Example C4-1 -2 4-Methyl-2-(3-trifluoromethanesulfonyloxy-phenyl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Under argon at -30 0 C, 2.6-lutidine (350 mg, 3.27 mmol) was added dropwise to a solution of 2- (3-hydroxy-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-1 -1 (500 mg, 1 .90 mmol) in dry methylene chloride.
  • Example C4-1 -3 / PKE 386 4-Methyl-2-(3-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(3-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-l -2 and thiophene-2-boronic acid; white solid (30%).
  • Example C4-1 -4 / PKE 383 2-(3-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(3-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-1 -2 and benzo[b]thiophene-2-boronic acid; white solid (65%).
  • Example C4-1 -5 / PKE 387 2-(3-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(3-benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-M; pale yellow solid (93%).
  • Example C4-2-1 / PKE 445 2-(4-Hydroxy-phenyl)4-methyl-thiazole-5-carboxylic acid ethyl este: Method as for Al -1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid ethyl ester and 4-hydroxyphenyl-boronic acid; white solid (43%).
  • Example C4-2-2 4-Methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for C4-l -2, using 2-(4-hydroxy-phenyl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4- 2-1 ; white solid (81 %).
  • Example C4-2-3 / PKE 384 4-Methyl-2-(4-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-2 and thiophene-2-boronic acid; white solid (57%).
  • Example C4-2-4 / PKE 442 2-(4-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-2 and benzo[b]thiophene-2-boronic acid; beige solid (71 %).
  • Example C4-2-5 / PKE 408 2-Biphenyl4-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 4-biphenyl boronic acid; white solid (45%).
  • Example C4-2-6 / PKE 457 4-Methyl-2-(4-naphthalen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-2 and naphthalene-2-boronic acid; white solid (59%).
  • Example C4-2-7 / PKE 400 4-Methyl-2-(4-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-3; yellow solid (22%).
  • Mp 21 1 °C-21 3°C.
  • Example C4-2-8 / PKE 458 2-(4-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(4-benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic ethyl ester C4-2-4; yellow solid (96%).
  • Example C4-2-9 / PKE 432 2-Biphenyl4-yl-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-Biphenyl-4-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-2-5; white solid (82%).
  • Example C4-2-1 0 / PKE 460 4-Methyl-2-(4-naphthalen-2-yl-phenyl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4-naphthalen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-6; yellow solid (86%).
  • Example C4-3-1 2-(2-Hydroxy-phenyl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 2-hydroxyphenyl-boronic acid; beige solid.
  • Example C4-3-2 4-Methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for C4-1 -2, using 2-(2-hydroxy-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-3-1 ; pale yellow oil (27% for two steps).
  • Example C4-3-3 / PKE 409 2-(2-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-2 and benzo[b]thiophene-2-boronic acid; light pink solid (94%).
  • Example C4-3-4 / PKE 434 2-(2-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(2-benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-3-3; white solid (67%).
  • Example C4-3-5 / PKE 41 0 4-Methyl-2-(2-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-2 and thiophene-2-boronic acid; beige solid (88%).
  • Example C4-3-6 / PKE 435 4-Methyl-2-(2-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid: thod as for B2-1 , using 4-methyl-2-(2-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-5; white solid (55%).
  • Example C4-3-7 / PKE 433 4-Methyl-2-(4 '-methyl-biphenyl-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-2 and 4-tolyl boronic acid; white solid (82%).
  • Example C4-3-8 / PKE 436 4-Methyl-2-(4 '-methyl-biphenyl-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4 '-methyl-biphenyl-2-yl)-thiazole-5-carboxylic acid ethyl ester C4-3-7; white solid (95%).
  • Example C4-4-1 2-(6-Hydroxy-pyridin-3-yl)4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 2-hydroxypyridine-5-boronic acid pinacol ester; brown solid (55%).
  • Example C4-4-2 4-Methyl-2-(6-trifluoromethanesulfonyloxy-pyridin-3-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for C4-1 -2, using 2-(6-hydroxy-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4 ⁇ -l ; white solid (14%).
  • Example C4-4-3 / PKE 437 2-(6-Benzo[b]thiophen-2-yl-pyridin-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 4-methyl-2-(6- trifluoromethanesulfonyloxy-pyridin-3-yl)-thiazole-5-carboxylic acid ethyl ester C4-4-2; yellow solid (36%).
  • Example C4-4-4 / PKE 441 2-(6-Benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(6-benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4-4-3; yellow solid (90%).
  • Example C4-5-1 5-Bromo-thionicotinamide: o,o'-Diethyl-dithiophosphate (3.3 ml, 21 .4 mmol) is added to a mixture of 5-bromonicotinonitrile (730 mg, 4.3 mmol) in pyridine (1 5 ml) and water (3 ml). Reac- tion mixture is stirred at 90 0 C in a sealed tube for 72 h. Solvents are evaporated to dryness. Residue is dissolved back in EtOAc, and washed three times with water. Organic layer is dried (Na 2 SO 4 ), filtered and solvents are evaporated to give a yellow solid.
  • Example C4-5-2 / PKE 459 2-(5-Bromo-pyridin-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: To a mixture of 5-bromo-thionicotinamide C4-5-1 (1 00 mg, 0.46 mmol) in absolute ethanol (3 ml) is added ethyl-2-chloroacetoacetate (0.064 ml, 0.46 mmol). Reaction mixture is heated up to 1 80 0 C in a microwave apparatus for 1 5 min under pressure (20 bars). Solvents are evaporated to dryness.
  • Example C4-5-3 / PKE 471 2-(5-Benzo[b]thiophen-2-yl-pyridin-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-pyridin-3-yl)-4- methyl-thiazole-5-carboxylic acid ethyl ester C4-5-2; white solid (48%).
  • Example C4-5-4 / PKE 473 2-(5-Benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4-5-3; yellow solid (1 00%).
  • Example C4-6-1 / PKE 444 2-Benzo[b]thiophen-2-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using benzo[b]thiophene-2-boronic acid and 2-bromo4-methyl-thiazole-5- carboxylic acid ethyl ester; beige solid (44%).
  • Example Dl -I / PKE 467 4-Methyl-2-[5-(thiophene-2-carbonyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: To a mixture of thiophene-2-carboxylic acid chloride (0.1 1 ml, 1 .0 mmol) and anhydrous ferric chloride (1 65 mg, 2.0 mmol) in dry dichloromethane (25 ml) was added 4-methyl-2-thiophen-2-yl- thiazole-5-carboxylic acid ethyl ester Bl -1 -1 (253 mg, 1 .0 mmol).
  • Example D2-1 -1 2-[(5-Bromo-thiophene-2-carbonyl)-amino]-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: A solution of ethyl-2-amino4-methyl-thiazole-5-carboxylate (430 mg, 2.3 mmol), 5-bromo-2- thiophene carboxylic acid (400 mg, 1 .9 mmol), 1 -hydroxybenzotriazole hydrate (320 mg, 2.3 mmol) and l -(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (445 mg, 2.3 mmol) in N 1 N- dimethylformamide (20 ml) was heated up to 60 0 C in a sealed tube for 1 8 h.
  • Example D2-1 -2 / PKE 496 2-[(5-Benzo[b]thiophen-2-yl-thiophene-2-carbonyl)-amino]4-methyl- thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2- [(S-bromo-thiophene ⁇ -carbonylJ-aminoH-methyl-thiazole-S-carboxylic acid ethyl ester D2-1 -1 ; yellow solid (1 00%).
  • Example D2-2 / PKE 497 2-[(5-Benzo[b]thiophen-2-yl-thiophene-2-carbonyl)-methyl-amino]4-methyl- thiazole-5-carboxylic acid ethyl ester: To a suspension of 2-[(5-benzo[b]thiophen-2-yl-thiophene-2- carbonyl)-amino]4-methyl-thiazole-5 ⁇ arboxylic acid ethyl ester D2-1 -2 (1 00 mg, 0.23 mmol) in N 1 N- dimethylformamide (5 ml) under argon at room temperature was added sodium hydride (60% dispersion in mineral oil, 1 3 mg, 0.32 mmol).
  • Reaction mixture was stirred at room temperature for 20 minutes, and methyl iodide (0.044 ml, 0.71 mmol) was then added. Reaction mixture was heated up to 60 0 C for 3 h. Saturated aqueous NH 4 CI was added, precipitate was filtered off, washed with saturated NaHCO 3 solution, and dried under vacuum to give a yellow solid (73%).
  • Example D2-3 / PKE 498 2-[(5-Benzo[b]thiophen-2-yl-thiophene-2-carbonyl)-methyl-amino]4-methyl- thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[(5-benzo[b]thiophen-2-yl-thiophene-2-carbonyl)- methyl-amino]-4-methyl-thiazole-5-carboxylic acid ethyl ester D2-2; yellow solid (75%).
  • Example D2-4 / PKE 469 4-Methyl-2-[(thiophene-2-carbonyl)-amino]-thiazole-5-carboxylic acid ethyl ester: Method as for D2-1 -1 , using 2-thiophene carboxylic acid and ethyl-2-amino4-methyl-thiazole-5- carboxylate; pink solid (50%).
  • Example E2-1 -1 5-Bromo-[ l ,3,4]thiadiazole-2-carboxylic acid ethyl ester: Method as for Al -1 -1 , using 5- amino-[ l ,3,4]thiadiazole-2-carboxylic acid ethyl ester; white solid (82%).
  • Example E2-2/ PKE 455 2-[2,2']Bithiophenyl-5-yl-[l,3,4]thiadiazole: Method as for Bl-1-1, using 2,2'- bithiophene-5-boronic acid and 5-Bromo-[l,3,4]thiadiazole-2-carboxylic acid ethyl ester E2-1-1; grey flakes (36%).
  • Example E2-3-1 2-Bromo-5-methyl-[l,3,4]thiadiazole: Method as for Al -1-1, using 5-methyl- [1 ,3,4]thiadiazol-2-ylamine; white solid (48%).
  • Example E2-3-2/ PKE 464 2-[2,2']Bithiophenyl-5-yl-5-methyl-[l,3,4]thiadiazole: Method as for Bl-1-1, using 2,2'-bithiophene-5-boronic acid and 2-bromo-5-methyl-[l,3,4]thiadiazole E2-3-1; greenish flakes (50%).
  • Example E2-4-1 2-Bromo-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Al -1-1, using 2-amino-5- trifluoromethyl-[l,3,4]thiadiazole; yellow oil (43%). 13 C NMR (CDCI 3 ) ⁇ 142.7, 120.2, 116.6.
  • Example E2-4-2/ PKE 504 2-[2,2']Bithiophenyl-5-yl-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Bl-1-1, using 2,2'-bithiophene-5-boronic acid and 2-bromo-5-trifluoromethyl-[l,3,4]thiadiazole E2-4-1; greenish solid (20%).
  • MS (ESI + ) m/z: 319.1 [M + H] + . Mp: 175°C-177°C.
  • Example E2-4-3 2-Thiophen-2-yl-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Bl-1-1, using thio- phene-2-boronic acid and 2-bromo-5-trifluoromethyl-[l,3,4]thiadiazole E2-4-1; yellow solid (44%).
  • MS (ESI + ) m/z: 236.9 [M + H] + .
  • Example E2-4-4 2-(5-Bromo-thiophen-2-yl)-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Bl-1-2, using 2-thiophen-2-yl-5-trifluoromethyl-[l,3,4]thiadiazole E2-4-3; white solid (60%).
  • Example E2-4-5 / PKE 508 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-5-trifluoromethyl- [l,3,4]thiadiazole: Method as for Bl-2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo- thiophen-2-yl)-5-trifluoromethyl-[ l ,3,4]thiadiazole E2-4-4; greenish solid (45%).
  • Example E3-1 -1 5-Bromo-2-methyl-benzoic acid ethyl ester: A solution of 5-bromo-2-methylbenzoic acid (200 mg, 0.93 mmol) and concentrated sulphuric acid (0.20 ml) in absolute ethanol (1 0 ml) was heated up to reflux for 1 5 h. Solvents were evaporated. Residue was dissolved back in EtOAc and washed with NaHCO 3 solution and brine. Organic layers were dried (MgS04), and concentrated under vacuum to give a yellow oil (97%).
  • Example E3-2 / PKE 531 5-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid: Method as for B2-1 , using 5- [2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester E3-1 -2; pale yellow solid (86%).
  • Example E3-3-1 2-Methyl-5-thiophen-2-yl-benzoic acid ethyl ester: Method as for Bl -1 -1 , using thio- phene-2-boronic acid and 5-bromo-2-methyl-benzoic acid ethyl ester E3-1 -1 ; orange oil (78%).
  • Example E3-3-2 5-(5-Bromo-thiophen-2-yl)-2-methyl-benzoic acid ethyl ester: Method as for Bl -1 -2, using 2-methyl-5-thiophen-2-yl-benzoic acid ethyl ester E3-3-1 ; orange oil (98%).
  • Example E3-4 / PKE 539 5-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-2-methyl-benzoic acid: Method as for B2-1 , using 5-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-2-methyl-benzoic acid ethyl ester E3-3-3; white solid (93%).
  • Example E3-5 / PKE 503 4-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester: Method as for Bl - 1 -1 , using 2,2'-bithiophene-5-boronic acid and 4-bromo-2-methyl-benzoic acid ethyl ester; yellowish solid (68%).
  • Example E3-6 / PKE 502 4-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid: Method as for B2-1 , using 4- [2,2 ']bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester E3-5; yellow solid (83%).
  • MS (ESI + ) m/z: 301 .1 [M + H] + . Mp: 305°C-308°C.
  • Example E4-1 -2 4-Methyl-5-thiophen-2-yl-1 ,3-thiazole-2-carboxylic acid ethyl ester: Method as for Bl -I - 1 , using thiophene-2-boronic acid and 5-bromo4-methyl-thiazole-2-carboxylic acid ethyl ester E4-1 -1 ; yellow solid (46%).
  • Example E4-1-3 5-(5-Bromo-thiophen-2-yl)4-methyl-l,3-thiazole-2-carboxylic acid ethyl ester: Method as for Bl-I -2, using 4-methyl-5-thiophen-2-yl-thiazole-2-carboxylic acid ethyl ester E4-1-2; yellow solid (96%).
  • Example E4-1-4/ PKE 538 5-(5-Benzo[b]thiophen-2yl-thiophen-2-yl)-4-methyl-thiazole-2-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 5-(5-Bromo-thiophen-2-yl)-4- methyl-thiazole-2-carboxylic acid ethyl ester E4-1-3; yellow solid (94%).

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Abstract

Compounds according to the invention are 2-(3-Q5-4-Q4-5-Q3-thien-2-yl)-4-R3-5-R2-1,3-thiazol, 2-(3-Q5-4-Q4- 5-Q3-fur-2-yl)-4-R3-5-R2-1,3-thiazol, 2-(3-Q5-4-Q4-5-Q3-thien-2-yl)-5-R2-1,3,4-thiadiazol, 2-(3-Q5-4-Q4-5-Q3-fur-2-yl)-5-R2-1,3,4-thiadiazol, 2-(2-Q3-4-Q5-1,3-thiazol-5-yl)4-R3-5-R2-1,3-thiazol, 2-(2-Q3-4-Q5-1,3-oxazol-5-yl)4- R3-5-R2-1,3-thiazol, 2-(4-Q4-5-Q3-1,3-thiazol-2-yl)4-R3-5-R2-1,3-thiazol, 2-(4-Q4-5-Q3-1,3-oxazol-2-yl)4-R3-5- R2-1,3-thiazol, 2-(4-Q5-5-Q4-3-Q2-thien-3-yl)4-R3-5-R2-1,3-thiazol, 2-(4-Q5-5-Q4-3-Q2-fur-3-yl)4-R3-5-R2-1,3-thiazol, 2-benzo[b]thiophen-2-yl4-R3-5-R2-1,3-thiazol, 2-benzo[b]furan-2-yl4-R3-5-R2-1,3-thiazol, 2-(3-Q5-4- Q4-5-Q3-thien-2-yl)-6-Z2-1,3-benzo[d]thiazol, 2-(3-Q5-4-Q4-5-Q3-thien-2-yl)-6-Z2-1,3-benzo[d]oxazol, 2-(2-Z5-3- Z6-4-Z7-phenyl)4-R3-5-R2-1,3-thiazol, 2-(5-Z8-6-Z7-3-pyridyl)4-R3-5-R2-1,3-thiazol, 2-(3-R3-4-R2-phenyl)-3-Q5- 4-Q4-5-Q3-thiophen, 2-(3-R3-4-R2-phenyl)-3-Q5-4-Q4-5-Q3-furan, N-Z9-N-(4-R3-5-R2-1,3-thiazol-2-yl)-3-Q5-4-Q4- 5-Q3-thien-2-yl-carboxamid, N-Z9-N-(4-R3-5-R2-1,3-thiazol-2-yl)-3-Q5-4-Q4-5-Q3-fur-2-yl-carboxamid, 5-(3-Q5-4- Q4-5-Q3-thien-2-yl)4-R4-2-R2-1,3-thiazol, 5-(3-Q5-4-Q4-5-Q3-fur-2-yl)4-R4-2-R2-1,3-thiazol, 2-(3-Q5-4-Q4-5-Q3- thien-2-yl)-3-R4-4-R3-5-R2-thiophen, 2-(3-Q5-4-Q4-5-Q3-fur-2-yl)-3-R4-4-R3-5-R2-thiophen; where R2, R3, R4, Q2, Q3, Q4, Q5, Z2, Z5, Z6, Z7, Z8 and Z9 are as defined in the description. Said compounds are particularly advantageous for treating and/or preventing influenza type A and/or influenza type B infections in humans, mammals and/or birds, and for treating and/or preventing respiratory syncytial virus infections in humans, mammals and/or birds.

Description

SMALL MOLECULE INHIBITORS OF INFLUENZA A AND B VIRUS AND RESPIRATORY SYNCYTIAL VIRUS REPLICATION
FIELD OF THE INVENTION
The present invention relates to small molecules inhibiting the replication of influenza A and B virus and respiratory syncytial virus (RSV), and the use of such compounds for treating influenza A and B and RSV infections, in humans, mammals and birds.
BACKCROUND OF THE INVENTION
Influenza viruses are negative-stranded RNA viruses that cause yearly epidemics as well as recurring pandemics, resulting in high numbers of human cases and severe economic burden. In addition to the well-known pandemic influenza A viruses (such as the 1 91 8 "Spanish" flu or H5N 1 ), both type A and B viruses contribute greatly to the annual recurring epidemics that cause the vast majority of human cases and medical cost. The WHO recommends an annual vaccination against circulating influenza A (FIuA) and B (FIuB) strains. However, current vaccines confer incomplete protection against epidemic influenza. To date, only the neuraminidase inhibitors oseltamivir (Tamiflu™) and zanamivir (Relenza™) are available as antiviral treatment against both virus types. However, there is a growing fear within the medical community about the rapidly growing emergence of influenza strains resistant to both drugs. The older adamantane drugs are not effective against FIuB and the global spread of influenza viruses resistant to oseltamivir demonstrate the limitations of the neuraminidase inhibitors. A recent epidemiological survey in the U.S. found 98.5% of the H l N l isolates tested resistant to oseltamivir.
Human respiratory syncytial virus (RSV) is a negative-sense, single-stranded RNA virus of the family Pa- ramyxoviridae, and is the major cause for respiratory tract illnesses during infancy and childhood such as bronchiolitis and pneumonia. There is currently no vaccine available. Treatment is mainly limited to supportive care, including oxygen. Palivizumab (Synagis™) is used as a prophylactic drug in prevention of respiratory RSV infections for infants with a high risk of infection. Ribavirin has been used for treating RSV infections, but showed limited effectiveness.
Thus, new improved and alternative antiviral agents against both influenza A and B virus types and RSV are urgently needed. OBJECTS OF THE INVENTION
One object of the invention is to provide new, improved and/or alternative influenza and RSV antiviral com pounds.
Another object of the invention is to obviate or mitigate disadvantages of influenza antiviral agents and RSV antiviral agents known from the state of the art.
It is a further object of the present invention to provide a novel method for identifying small molecules that are active as protein-protein interaction disruptors.
These and other objects are achieved by a compound, a compound as a medicament, a compound for treating influenza type A and/or influenza type B and/or RSV infections in humans, mammals and/or birds, the use of a compound for the manufacture of a medicament for the treatment of influenza type A and/or influenza type B and/or RSV infections in humans, mammals and/or birds, and a pharmaceutical composition comprising such a compound, as well as a method for identifying small molecule disruptors of protein-protein interactions, according to the independent claims. Advantageous embodi- ments are given in the dependent claims.
SUMMARY OF THE INVENTION
A compound according to the invention is 2-(3-Q54-Q4-5-Q3-thien-2-yl)4-R3-5-R2-l ,3-thiazol, 2-(3-Q54-Q4- 5-Q3-fur-2-yl)-4-R3-5-R2-l ,3-thiazol, 2-(3-Q54-Q4-5-Q3-thien-2-yl)-5-R2-l ,3,4-thiadiazol, 2-(3-Q5-4-Q4-5-Q3-fur- 2-yl)-5-R2-l ,3,4-thiadiazol, 2-(2-Q3-4-Q5-l ,3-thiazol-5-yl)4-R3-5-R2-l ,3-thiazol, 2-(2-Q34-Q5-l ,3-oxazol-5-yl)4- R3-5-R2-l ,3-thiazol, 2-(4-Q4-5-Q3-l ,3-thiazol-2-yl)4-R3-5-R2-l ,3-thiazol, 2-(4-Q4-5-Q3-l ,3-oxazol-2-yl)4-R3-5- R2-l ,3-thiazol, 2-(4-Q5-5-Q4-3-Q2-thien-3-yl)4-R3-5-R2-l ,3-thiazol, 2-(4-Q5-5-Q4-3-Q2-fur-3-yl)4-R3-5-R2-l ,3- thiazol, 2-benzo[b]thiophen-2-yl4-R3-5-R2-l ,3-thiazol, 2-benzo[b]furan-2-yl4-R3-5-R2-l ,3-thiazol, 2-(3-Q54- Q4-5-Q3-thien-2-yl)-6-Z2-l ,3-benzo[d]thiazol, 2-(3-Q54-Q4-5-Q3-thien-2-yl)-6-Z2-l ,3-benzo[d]oxazol, 2-(2-Z5-3- Z64-Z7-phenyl)4-R3-5-R2-l ,3-thiazol, 2-(5-Z8-6-Z7-3-pyridyl)4-R3-5-R2-l ,3-thiazol, 2-(3-R34-R2-phenyl)-3-Q5- 4-Q4-5-Q3-thiophen, 2-(3-R34-R2-phenyl)-3-Q54-Q4-5-Q3-furan, N-Z9-N-(4-R3-5-R2-l ,3-thiazol-2-yl)-3-Q54-Q4- 5-Q3-thien-2-yl-carboxamid, N-Z9-N-(4-R3-5-R2-l ,3-thiazol-2-yl)-3-Q54-Q4-5-Q3-fur-2-yl-carboxamid, 5-(3-Q54- Q4-5-Q3-thien-2-yl)4-R4-2-R2-l ,3-thiazol, and 5-(3-Q54-Q4-5-Q3-fur-2-yl)4-R4-2-R2-l ,3-thiazol, 2-(3-Q54-Q4-5-
Q -thien-2-yl)-3-R 4-R -5-R -thiophen, 2-(3-Q 4-Q -5-Q -fur-2-yl)-3-R 4-R -5-R -thiophen, represented by formulas (Xl) to (XXIII)
Figure imgf000004_0001
Figure imgf000005_0001
where X is sulfur or oxygen; where R2 is selected from a group consisting of: hydrogen, cyano, formyl, acetyl, ethylcarbonyl, propylcarbonyl, isopropylcarbonyl, carboxy, carboxymethyl, methoxycarbonyl, eth- oxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert- butoxycarbonyl, methoxycarbonylmethyl, methoxycarbonylethyl, ethoxycarbonylmethyl, methoxycarbon- ylethyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trihalogenomethyl, difluoromethyl, dihalogenomethyl, fluoromethyl, halogenomethyl, phenyl, 1 -naphthyl, 2-naphthyl, aryl, hydroxy methyl, 1 -hydroxyethyl, 2-hydroxyethyl, carbamoyl, hydroxycarbamoyl, methylcarbamoyl, ethyl- carbamoyl, propylcarbamoyl, isopropylcarbamoyl, butylcarbamoyl, isobu-tylcarbamoyl, tert- butylcarbamoyl, (l -(2-thienylcarbonyl)-pyrazol-5-yl), (l -(m-trifluoromethyl-benzoyl)-pyrazol-5-yl), (l -(tert- butylcarbonyl)-pyrazol-5-yl), ( 1 -ethy l-tetra hyd ro py rro l-2-y I )-m ethy l-ca rba m oy I , (l -methyl-imidazol-4-yl)- sulfonamidomethyl, (2-morpholinoethoxy)-carbonyl, (2-morpholinoethyl)-carbamoyl, (3-(imidazol-l -yl)- propyl)-carbamoyl, (3-morpholinopropyl)-carbamoyl, (4-(p-fluorophenyl)-2,3,5,6-tetrahydropyrazin-l -yl)- carbonyl, (tetrahydropyrrol-l -yl)-carbonyl, 2-benzofuranyl, benzamidomethyl, cyclopropylamidomethyl, C1 -C4 alkylami-domethyl, benzamidoethyl, cyclopropylamidoethyl, C1 -C4 alkylamidoethyl, phenylsul- fonamidomethyl, cyclosulfonamidomethyl, C1 -C4 alkylsulfonamidomethyll, phenylsul-fonamidoethyl, cyclosulfonamidoethyl, and C1 -C4 alkylsulfonamidoethyl, 1 W-tetrazol-5-yl, 2W-tetrazol-5-yl, 1 W-I -Zl O- tetrazol-5-yl, 2/-/-1 -Zl 0-tetrazol-5-yl, where Zl O is selected from a group consisting of: methyl, C2-C4 alkyl, and substituted alkyl chains; where R3 is selected from a group consisting of: hydrogen, formyl, acetyl, ethylcarbonyl, propylcarbonyl, isopropylcarbonyl, carboxy, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxy-carbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trihalogenomethyl, difluromethyl, di- halo-genomethyl, fluoromethyl, halogenomethyl, hydrazinocarbonyl, m,p-difluorophenyl, m,p- dihalogenophenyl, p-(trifluoromethyl)-phenyl, p-(trihalogenomethyl)-phenyl, p-chlorophenyl, p- halogenophenyl, and p-nitrophenyl; where R4 is selected from a group consisting of: methyl, C2-C4 alkyl; where Q2 is selected from a group consisting of: hydrogen, 2-thienyl, 5-acetyl-thien-2-yl, and aryl; where Q3 is selected from a group consisting of: hydrogen, phenyl, 1 -naphthyl, 2-naphthyl, methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, tert-butyl, bromo, halogeno, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-thienyl, 3- thienyl, 2-furyl, 3-furyl, 4-pyrazolyl, benzo[b]furan-2-yl, benzo[b]thiophen-2yl, 3-quinolyl, 1 -isoquinolyl, 4- isoquinolyl, 2-indolyl, 5-pyrimidinyl, (2-thienyl)-carbonyl, l -(tert-butoxycarbonyl)-indol-2-yl, 1 ,2-oxazol-5-yl, 1 ,3-benzodioxol-5-yl, l ,3-thiazol-2-yl, l -methylpyrazol4-yl, 2,4-dimethyl-l ,3-thiazol-5-yl, 4-methyl-2-phenyl- l ,3-thiazol-5-yl, 4-phenyl-thien-2-yl, 5-(2-thienyl)-thien-2-yl, 3-methyl-thien-2-yl, 3-ethyl-thien-2-yl, 4-methyl- thien-2-yl, 4-ethyl-thien-2-yl, o-Q7-phenyl, m-Q7-phenyl, p-Q7-phenyl, o,m-bis(Q7)-phenyl, o,p-bis(Q7)-phenyl, m,p-bis(Q7)-phenyl, 5-Q7-benzo[b]thiophen-2-yl, 5-Q7-benzo[b]furan-2-yl, 5-Q7-benzo[b]thiophen-3-yl, 5-Q7- benzo[b]furan-3-yl, 5-Q7-benzo[b]thiophen-5-yl, 5-Q7-benzo[b]furan-5-yl, 6-Q7-benzo[b]thiophen-3-yl, 6-Q7- benzo[b]furan-3-yl, 6-Q7-benzo[b]thiophen-5-yl, 6-Q7-benzo[b]furan-5-yl, 6-Q7-naphth-2-yl, and 5-Q7-thien- 2-yl, wherein Q7 is selected from a group consisting of: hydroxy, trifluoromethoxy, trifluoromethoxy, tri- halogenomethoxy, difluoromethoxy, dihalogenomethoxy, fluoromethoxy, halogenomethoxy, trifluoro- methyl, trihalogenomethyl, difluoromethyl, dihalogenomethyl, fluoromethyl, halogenomethyl, formyl, acetyl, chloro, fluoro, halogeno, cyano, nitro, methyl, ethyl, C3-C4 alkyl, methoxy, ethoxy, and C3-C4 alkoxy; where Q4, Q5, and Z9 are independently selected from a group consisting of: hydrogen, methyl, ethyl, C3-C4 alkyl, and phenyl; where Z2 is selected from a group consisting of: hydrogen, methyl, ethyl, C3-C4 alkyl, methoxy, ethoxy, and C3-C4 alkoxy; where Z5, Z6, and Z7 are independently selected from a group consisting of: hydrogen, phenyl, p-tolyl, 1 -naphthyl, 2-naphthyl, 2-thienyl, 2-furyl, benzo[b]thiophen-2-yl, benzo[b]furan-2-yl, and hydroxy; and where Z8 is selected from a group consisting of: bromo. halogeno, benzo[b]thiophen-2-yl, and benzo[b]furan-2-yl; or a physiologically tolerable salt, solvate, or physiologically functional derivative thereof.
Said compounds according to the invention are particularly advantageous as a medicament.
The following are compounds according to the invention for which the medical indication is claimed: 1 - {4-Methyl-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazol-5-yl}ethan-l -one; 4-(4-nitrophenyl)-2-[5-(2-thienyl)-2- thienyl]-l ,3-thiazole; 4-(4-chlorophenyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole; 2-[5-(2-thienyl)-2-thienyl]-4- [4-(trifluoromethyl)phenyl)-l ,3-thiazole; 4-(3,4-difluorophenyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole; 4- (tert-butyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole; (2-thienyl)(5-{2-[5-(2-thienyl)-2-thienyl]-4-methyl-l ,3- thiazol-5-yl}-l H-pyrazol-1 -yl)methanone; 1 -(5-{2-[5-(2-thienyl)-2-thienyl]-4-methyl-l ,3-thiazol-5-yl}-l H- pyrazol-1 -y I )-2 , 2-d i m ethy I p ro pa n- 1 -one; (5-{2-[5-(2-thienyl)-2-thienyl]4-methyl-l ,3-thiazol-5-yl]-l H-pyrazol- l -yl)[3-(trifluoromethyl)phenyl]methanone; 5-[5-(4-benzo[b]furan-2-yl-l ,3-thiazol-2-yl)thien-2-yl]isoxazole; methyl [2-(5-isoxazol-5-ylthien-2-yl)-5-methyl-l ,3-thiazol4-yl]acetate; 2-[5-(4-phenyl-l ,3-thiazol-2-yl)-2- thienyl]pyridine; 2-{5-[4-(tert-butyl)-l ,3-thiazol-2-yl]-2-thienyl}pyridine; ethyl 2-(5-isoxazol-5-ylthien-2-yl)-4- methyl-1 ,3-thiazole-5-carboxylate; 4-methyl-2-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-l ,3-thiazole-5- carboxylic acid; N-{[2-(2-thienyl)-l ,3-thiazol4-yl]methyl} benzenesulfonamide; N-{[2-(2-thienyl)-l ,3- thiazol-4-yl]methyl} benzamide; l -[4-methyl-2-(2-thienyl)-l ,3-thiazol-5-yl]ethan-l -one; 4-methyl-5-(l H- pyrazol-5-yl)-2-(2-thienyl)-l ,3-thiazole; 4-methyl-5-[2-(methylthio)pyrimidin4-yl]-2-(2-thienyl)-l ,3-thiazole; cyclopropyl{5-[4-methyl-2-(2-thienyl)-l ,3-thiazol-5-yl]-l H-pyrazol-l -yl}methanone; N l -isopropyl-5-[4- methyl-2-(2-thienyl)-l ,3-thiazol-5-yl]-l H-pyrazole-1 -carboxamide; l -methyl-N-{[2-(2-thienyl)-l ,3-thiazol-4- yl]methyl}-l H-imidazole-4-sulfonamide; N-{[2-(2-thienyl)-l ,3-thiazol-4-yl] methyl} cyclopropanecarbox- amide; 4-methyl-2-(2-thienyl)-l ,3-thiazole-5-carbohydrazide; or a physiologically tolerable salt, solvate, or physiologically functional derivative thereof.
The above defined compounds according to the invention are particular advantageous for treating and/or preventing influenza type A and/or influenza type B infections in humans, mammals and/or birds; as well as for treating and/or preventing respiratory syncytial virus (RSV) infections in humans, mammals and/or birds.
The compound according to the invention can be used for the manufacture of a medicament for the treatment and/or prevention of influenza type A and/or influenza type B infections in humans, mam- mals and/or birds, and/or for the treatment and/or prevention of respiratory syncytial virus infections in humans, mammals and/or birds.
A pharmaceutical composition according to the invention comprises a compound according to the invention. Advantageously such a composition comprises one or more excipients.
A method according to the invention for identifying small molecules and/or peptidic inhibitors or dis- ruptors for protein-protein interactions (PPIs) comprises the steps of: a) selection of a proteome of interest; b) identification of potential target regions of PPI inhibitors; c) detection of drugable protein- binding domains; d) synthesis of peptides corresponding to the drugable protein-binding domains; e) testing of the synthesized peptides for their ability to bind the protein partner involved in the PPI; f) development of a binding assay for the identification of small molecules and/or peptidic inhibitors or disrupters based on the positively tested peptides; and g) high-throughput screening with the binding assay and selecting the active small molecules and/or peptidic inhibitors or disruptors.
Surprisingly, it was found that compounds in accordance with the present invention are able to inhibit protein-protein interaction of the PA and PBl subunits of the heterotrimeric viral RNA polymerase complex of both influenza virus types A and B, and thus are able to inhibit replication of influenza A and B virus. The viral polymerase subunit interaction domain turned out as an effective target for the new antiviral compounds, since correct assembly of the three viral polymerase subunits PBl , PB2 and PA is required for viral RNA synthesis and infectivity. Structural data for the entire trimeric complex is missing. Based on the crystal structure of a truncated FIuA PA in complex with the N-terminus of PBl it was established that the crucial PA interaction domain of PBl consists of a 3io-helix formed by amino acids (amino acids 5-1 1 ). The domain is highly conserved and virus type specific among both, influenza A and B viruses.
An Enzyme-Linked Immunosorbent Assay (ELISA) based screening assay and other assays are used to prescreen compounds according to the invention that show antiviral activity against influenza A and B viruses. Since they are effective against both virus types, such compounds represent an attractive alternative to neuraminidase inhibitors. Therefore, the present invention represents a major step toward a sorely needed, near-universal medicament against influenza virus, and one which, due to its protein- protein interaction domain target, will likely be less susceptible to the emergence of drug-resistant strains for which influenza is well known.
Furthermore it was found that compounds according to the invention are also able to inhibit replication of respiratory syncytial virus (RSV).
Thus the compounds according to the invention can be used as a medicament, particularly as an influenza virus and/or RSV replication inhibitor and an influenza and/or RSV preventive/therapeutic agent, respectively.
The object, characteristics, and advantages of the present invention as well as the idea thereof will be apparent to those skilled in the art from the descriptions given herein. It is to be understood that the embodiments and specific examples of the invention described herein below are to be taken as preferred examples of the present invention. These descriptions are only for illustrative and explanatory purposes and are not intended to limit the invention to these embodiments or examples. Other aspects and features of the present invention will become apparent to those ordinarily skilled in the art upon review of the following description of specific embodiments of the invention in conjunction with the accompanying figures. It is further apparent to those skilled in the art that various changes and modifications may be made based on the descriptions given herein within the intent and scope of the present invention disclosed herein.
DETAILED DESCRIPTION OF THE INVENTION
Influenza Type A and B: Therapeutic target
In the international patent application with the title "Influenza A and B virus replication-inhibiting peptides" No. PCT/EP2009/055632, filed on 8 May 2009, novel peptides containing for example amino acid sequences from both virus types A and B, are described. The content of said application is hereby included by reference in its entirety. Surprisingly, it was found that those novel peptides bind to PA sub- units of both types of influenza A and B. Among said novel peptides, chimeric peptides, containing amino acid sequences from both virus types A and B, were identified which not only bind to both PA subunits, but also decrease the viral polymerase activity and the spread of virus in cell culture for both influenza A and B. In the following the findings concerning the binding of said novel peptides are described in order to further specify the inhibition target of the small molecules compounds according to the present invention.
It should be noted that all amino acids are preferably indicated by the IUPAC one letter code in the present application. Whenever three letter codes are used, they are also in accordance with IUPAC. The letter X is used to indicate a wildcard/variable or other amino acid at a certain position.
It has been found that the crucial PA interaction domain of PBl consists of a 3io-helix formed by amino acids X5 to Xn. This domain is highly conserved and type-specific among both influenza A and B viruses (Figure Ia). Additionally, FIuB PBl was able to bind to FIuA PA when these 25 amino acids were exchanged with the FIuA PBl sequence (Figure 2).
Table Ia shows the inhibitory concentrations of FluA/FluB-derived peptides determined by competitive ELISA. Competitor peptides (0.048 to 300OnM) were mixed with cell extracts containing HA-tagged PA from either FIuA or FIuB. Table 1 lists 12 competitive peptides. The first peptide PBIM5A is the FIuA wild type, the second row shows the FIuB wild type. For the peptides of rows 3 to 8 letters indicate FIuB specific amino acids. Rows 9 to 12 list further competitive peptides with amino acids at position 6 being neither FIuA nor FIuB specific. Standard deviation is indicated in parenthesis. Asterisks indicate highest concentrations of peptides used without reaching 50% inhibition. Further competitive peptides which are not listed in the table but have effectively reached 50% inhibition at low peptide concentra- tions are PB1 M5AT6I , PB1M5AT6L and PB1 M5AT6V, Peptides with slightly lower inhibition activity are PBl 1-I5A16A and PBl I-I5AT6M which are also not shown in Table Ia.
Table Ia: inhibitory concentrations of FluA/FluB-derived peptides determined by competitive ELISA
IC50(ITM)
Competitive peptide PA s'F!u/V! PA (FIuB)
PB1,- MDVNPTLLFLKVPAQ 43.32*+/- 5.31} >3000-
PBI1. s B .^I..1'F..ID..1. >3000* 45.0 (+/-12.5)
PBI1. 5 AjJsN1 V3! 1.1 ■ .f-ΪX.. X. 6.69 (+/- 1.73) >30G0*
PBI1.5 As_:0ικi:0 >3000" >3OOG*
PBI1, 5. Aθ2M V31 ,m, 12.96 *+/- 3.98} >30O0"
PBI1. S A rev L7F" -S? F 7.51 !>/- 071 ) 3450 (+/-81.5)
PBI1 ...... F ........ >3000* >3000'
PB1,. .....?;...,,.,.. 21.84(+M .48) 107.1 {+/-31.3}
PBI1, 5 A τ$κ .....F......... 2.84 (+/- 048) 750.4 (+/-249.6)
F1BI1 s A raw ..... W 3.40 (+/-0.51) 628.3 {+/-389.1)
PBI1- s Ar8M .H 282.16 (+/-34.04} >3000'
PBI1 5 Aτ5C .C... 4358 (+/- 5.87) >3000"
' highest concentration of competitive peptide used A comprehensive and qualitative overview on further peptides with high inhibitory activity is provided in Table 1 b. In the table the amino acid sequences at positions X5 to Xi0 of wild type A mutants are indicated.
Table I b: qualitative overview of further preferred peptides
Figure imgf000010_0001
In Table 1 c the amino acid sequences at amino acid residues X5 to Xi0 of wild type A mutants are indicated. Said peptides exhibit lower activities than the above mentioned peptides according to Tables I a and 1 b. Table I c: qualitative overview of further peptides
Figure imgf000010_0002
Based on the above presented information and results, it is clear for the person skilled in the art, that the synthesized or isolated influenza virus replication-inhibiting peptides interacting with the inhibition target for the small molecules compounds according to the invention comprise an amino acid sequence Of X5X6X7X8X9XiO, wherein X5 is P; X6 is T, Y, F, W, H, C, I, L, V, A or M; X7 is L or F; X8 is L, I, F or M; X9 is F, Y, W, H, L, R or S, and Xio is L, I or Y. Said amino acid sequence is at least 60 %, preferably at least 70%, more preferably at least 80% or 90% identical to the polypeptide according to the wild type PBI mA which is M DVN PTLLFLK. Within the aforementioned group of peptides, those peptides are preferred which comprising the amino acid sequence Of X6X7X8XgXi0, wherein X6 is T, Y, F, W, H, C, I, L or V; X7 is L or F; X8 is L or I; X9 is F, Y or W and Xi0 is L. Even more preferred according to certain embodiments are peptides that comprise the amino acid sequence Of X6X7, wherein X6 is T, Y, F, W, H, C, I, L or V and X7 is L or F.
Effective peptides advantageously comprise at least 1 1 residues Xi n, whereby preferably the proteins comprise the amino acid sequence M DVN PX6X7 LFLKVPAQ wherein X6 is selected from the group: T, Y, F, W, H. C, A, I, L, V or M and X7 is selected from the group L or F. A preferred peptide comprises an amino acid sequence elected from the group: MDVNPYFLFLKVPAQ, MDVNPYLLFLKVPAQ, MDVNPWLLFLKVPAQ or MDVNPFLLFLKVPAQ. According to further preferred embodiments the peptides com prise at least 1 5 residues Xi 1 5 according to the wild type PBl 1 1 5A but not the wild type sequence MDVNPTLLFLKVPAQ.
Table 2 shows the 50%-inhibitory concentrations (IC50) of FluA-derived PBl peptides determined by competitive ELISA. Peptide PBI 1 25A was immobilized on microwell plates and incubated with increasing concentrations of competitor peptides and cell extract containing HA-tagged PA of FIuA. Bound PA was detected by HA-specific antibodies as described above. Standard deviation is shown in parenthesis. Asterisks indicate highest concentrations of peptides used without detectable inhibitory effect. Grey boxes highlight amino acids that are part of the 3iO-helix, which comprises the core PA-binding region of PBl . Amino acids known to form hydrogen bonds with PA residues are represented in bold. The systematic truncation of the 25mer peptide comprising the PA-binding domain of PBl at the N- and C- terminus showed - based on the ELISA assay results - that i) the 25mer peptide can be truncated at the C- terminus until the first 14 or even 1 3 N-terminal amino acids remain without losing ability to inhibit the bound peptide-PA interaction. Truncation at the C-terminus down to the first 1 2 or even 1 1 amino acids resulted in peptides which still showed considerable activity. The systematic truncation showed further that ii) N-terminal truncation is not possible without major loss in inhibitory activity of the peptide.
Table 2: Inhibitory concentrations (IC50) of FluA-derived PBl peptides
FIuA-PBI peptides (aa) IC50 (nM)
3,0-iieiιx
1 -25 MDVN^f kI*FX»KVPAQNAISTTFPYT 1 .80 {+/- 0 49)
3 _25 — VNPfLXdPLKVPAQNAISTTFFYT 661 77 {+/- 22 08)
5 -25 FTOiFl^KVPAQNAISTTFPYT 483.20 (+/- 51.98)
7 -25 LLFLKVPAQNAISTTFPYT >300CT
9 -25 FLKVPAQNAISTTFPYT >3GGG*
11 -25 KVPAQNAISTTFPYT >3GOO*
Figure imgf000011_0001
i -16 MDVNJ^X&T&KVFAQN 45 86 (+/- 4.22)
1 -15 MDVNPTX-L(FI1KVPAQ 43 32 (+/- 5.31 )
1 -14 MDVNFXL&FX.KVPA 34 53 (+/- 2.19)
1 - 13 MDVWFfLl-FtKVP 138.17 (+/- 7.88) i -12 MDVNS^X&F&KV 643 93 {+/- 180 75} i -l i MDVN£>TIΛFI»K 899.53 (+/- 54.31 )
1 -10 MDVNPTLLFL >3000Λ
1-9 MDVNPTLLF >3G0CP
1 -8 MDVNPTLL >300(T
1 -7 MDVNPTL >3GQ(T i-δ MDVNFT >3GQ0*
* highest concentration of competitive peptide used Table 3 illustrates the inhibitory concentrations (IC50) of FluA-derived competitor peptides determined by ELISA. Peptide PBl 1.25A was again immobilized on microwell plates and incubated with increasing concentrations of competitor peptide and cell extract containing HA-tagged PA of FIuA. HA-specific antibodies detected bound PA. Standard deviations are shown in parenthesis. Asterisks indicate highest concentrations of peptides used without detectable inhibitory effect.
Table 3: Inhibitory concentrations (IC50) of FluA-derived PBl peptides
Competitive peptide IC50 in nM PBIM5A MDVNPTLLFLKVPAQ 43.32 (+/- 5.31)
PBI1.15AMIA ADVNPTLLFLKVPAQ 460.30 (+/- 27.85)
PB1115AD2A MAVNPTLLFLKVPAQ 209.17 (+/-44.62) PBI1-15 A V3A MDANPTLLFLKVPAQ 154.93 (+/- 18.18) PBIv15A N4A MDVAPTLLFLKVPAQ >3000* PBIv15A P5A MDVNATLLFLKVPAQ 2728.67 (+/-133.43) PBIM5ATSA MDVNPALLFLKVPAQ 701.87 (+/- 20.59) PBIM5A LJA MDVNPTΆLFLKVPAQ >3000X PBIM5A LSA MDVNPTLAFLKVPAQ >3QQG* PBIM5 A F9A MDVNPTLLALKVPAQ >3GQ0* PBIv16ALIOA MDVNPTLLFAKVPAQ >3000' PBIv15A K11A MDVNPTLLFLAVPAQ 1290.33 (+/-210.37) PB1vi5AV12A MDVNPTLLFLKΆPAQ 707.87 (+/-168.54) PBIv16A P13A MDVNPTLLFLKVΆAQ 257.93 ('+/- 3676) PBIv15AMID DDVNPTLLFLKVPAQ 1375.67 (+/-268.11) PBI1 f5AV3D MDDNPTLLFLKVPAQ >3000" PBIv15AhMD MDVDPTLLFLKVPAQ >3000" PBIv15APSD MDVNDTLLFLKVPAQ >3000X PB1visAT6D MDVNPDLLFLKVPAQ 2067.67 (+/- 584.98) PBI1-I5 A L7D MDVNPTDLFLKVPAQ >3QQ0* PB1v,sALBD MDVNPTLDFLKVPAO >3G00* PBIv15 A F9D MDVNPTLLDLKVPAO >3000' PBIv15ALIOD MDVNPTLLFDKVPAQ >3000< PB1115AK11D MDVNPTLLFLDVPAQ >3000" PBIv15 A V12D MDVNPTLLFLKDPAQ 2302.67 (+/- 280.39} PBIv15API 3D MDVNPTLLFLKVDAQ 1097.47 (+/-217.54)
highest concentration of competitive peptide used Figure 1 shows binding and inhibitory activity of PB1 1 -25AT6Y Based on Figure 1 the binding and inhibitory activity of peptides binding to the inhibition target with a focus on the preferred protein PBl i 25A16Y shall be illustrated in the following part of the description Figure I a shows in the upper panel the alignment of the consensus sequence of the N-terminal 25 ammo acids of FIuA and FIuB PBl , wherein the dotted box indicates the 3io-helιx comprising the core PA-binding domain of PBl and the FluA-specific and FluB-specific ammo acids are printed in bold letters Middle and lower panels show the alignment of the N-terminal 25 ammo acids of all available FIuA and FIuB sequences derived from PBl full length sequences provided by the NCBI influenza virus database
The binding of HA-tagged PA subunits from cell extracts to the immobilized peptides corresponding to the domains of FIuA PBl (PBI i 25A), FIuB PBl (PBI i 25B) or FIuA PBl T6Y (PBI i 25Aτ6γ) determined by ELISA is shown in Figure I b Signals using the cognate peptide and lysate were normalized to 1 Binding of the PA subunits to the control peptides was not observed Upper panels Western blot of the PA- containmg cell extracts used Molecular weights shown in kilodaltons
Figure I c provides graphic information on the structure of FIuA PBI i i s bound to FIuA PA T6 forms a hydrogen bond to a water molecule Molecular modeling suggests that the aromatic side chain in the mutant T6Y fits into a hydrophobic pocket and displaces the water molecule The polymerase inhibitory activity of PBI i 25-derιved CFP fusion proteins in FIuA and FIuB polymerase reconstitution assays is shown in Figure I d The activity in experiments containing all viral plasmids and Flag-CFP was set to 1 00%
Figure I e shows a plaque reduction assay using PBl i 25A-Tat, PBl 1 25AT6γ-Tat, PX-Tat (control peptide) with FIuA, FIuB and VSV (vesicular stomatitis virus) A H2O control was used to standardize the assay to 1 00% Note that PBl i 25B-Tat could not be tested due to insolubility Error bars represent standard deviations
Virus type-specific interaction of PA with PBl is illustrated in Figure 2 Figure 2a shows A/SC35M- and B/Yamagata/73-derιved PBl chimeras used in tests according to Figure 2 b Note that all PBl proteins were expressed with C-terminal HA-tags Figure 2b shows human 293T cells which were transfected with expression plasmids coding for the indicated PBl proteins and the C-terminally hexahistidme- tagged PA of FIuA (FluAPAH|S) Cell lysates were prepared 24 hours post transfection and subjected to immunoprecipitation (IP) using antι-HA (aHA) agarose Precipitated material was separated by SDS- PACE and analyzed by Western blot for the presence of either His- or HA-tagged polymerase subunits using appropriate antibodies Protein expression was controlled by analyzing equal amounts of cell lysate Molecular weights are shown in kilodaltons The 25-mer peptide, PBI i 25A, comprising a helical domain inhibits the polymerase activity and replication of FIuA, whereas the activity of FIuB polymerase is not affected In Figure 3 dual-binding properties of the FluA/B peptide chimera PBI i 2SAT6Y are illustrated in comparison to PBI 1 25A and PBI i 25B The Lower panels show peptides PBI 1 25A, PBI 1 25B or PBl 1 25A16Y immobilized on microwell plates and incubated with increasing concentrations of cell extract containing the indicated PA-HA from FIuA or FIuB strains Bound PA-HA was detected by HA-specific antibodies and peroxidase-labeled secondary antibodies Binding efficiency was quantified by measuring substrate conversion at 405 nm Standard deviations are indicated by error bars Experiments were repeated in triplicates Upper panels show analysis of corresponding amounts of cell lysate by Western blot controlled protein expression Molecular weights are shown in kilodaltons
Figure 4a shows CFP-PBl fusion proteins used in tests according to Figure 4b The complex formation of PBl 1 25-derιved CFP fusion proteins and HA-tagged PA of FIuA and FIuB is shown in Figure 4b Indicated proteins were expressed in human 293T cells and binding of the CFP fusion proteins was analyzed by immunoprecitation (IP) of PA using anti-HA agarose and subsequent immunoblotting (IB) Precipitated material was analyzed by Western blot using the indicated antibodies for the presence of either HA- tagged PA or CFP Molecular weights are shown in kilodaltons
Influenza Type A and B: Materials and Methods
Virus strains: For the infection experiments A/WSN/33 (H l N l ) according to Chanem et al (2007) and A/Thaιland/l (Kan-l )/2004 according to Chockephaibulkit et al (2005), B/Yamagat/73 according to Norton (1 987) and VSV (serotype Indiana) as described in Schwem mle (1 995) were used
Plasmid constructions: Plasmids pCA-Flag-CFP and pCA-PBl 1 25A-CFP, pCA-PBl -HA, the FIuA minireph- con plasmids and the expression plasmids for the FIuB minireplicon are described in Chanem (2007), Mayer (2007) and Pleschka (1 996) The FIuB minigenome expression plasmid, pPoll-lucRT_B, was obtained by cloning the firefly luciferase ORF (inverse orientation) flanked by the non-coding region of the segment 8 of the B/Yamagata/73 into the Sapl-digested plasmid pPoll-Sapl-Rib according to Pleschka (1 996) For the construction of pCA-PBl i 25B-CFP, a linker containing the first 25 codons of PBl (B/Yamagata/73) was cloned into the EcoRI/Notl sites of pCA-Flag-CFP plasmid, replacing the Flag- coding sequence with PBI 1 25B Site directed mutagenesis was carried out with pCA-PBl 1 25A-CFP to create the plasmid pCA-PBl i 25AT6γ-CFP The ORFs of PBl (B/Yamagata/73) and PA (A/SC35M, A/Thaιland/1 (KAN-I )/04, A/Vιetnam/1 203/04, BAamagata/73, B/Lee/40) were PCR amplified with sense primers containing an Notl site (FIuA strains) or a EcoRI site (FIuB strains) upstream of the initiation codon and antisense primers with a deleted stop codon followed by an Xmal site, a coding sequence for an HA-tag and a Xhol site The PCR products were cloned into a modified pCACCsvector (Schneider, 2003) digested either with EcoRI/Xhol or Notl/Xhol, resulting in pCA-PBl -HA or pCA-PA- HA plasmids, coding for C-terminal tagged versions of the polymerase subunits To obtain the pCA- PAvsc35M-Hιs plasmid, pCA-PA^scsswrHA was digested with Xmal/Xhol and the HA coding sequence was replaced by a 6xH is-l inker. The A/B-chimeric expression plasmids were obtained by assembly PCR using the pCAPBl -HA plasmids of SC35M and B/Yamagata/73 and by cloning the resulting PCR product in pCA-PBl B/Yamagata/73-HA digested with EcoRI/EcoRV.
Reconstitution of the influenza virus polymerase activity: H EK293T cells were transiently transfected with a plasmid mixture containing either FIuA- or FluB-derived PBl -, PB2-, PA- and NP-expression plasmids, polymerase I (Pol l)-driven plasmid transcribing an influenza A or influenza B virus-like RNA coding for the reporter protein firefly luciferase to monitor viral polymerase activity and with expression plasmids coding for the indicated CFP fusion proteins. Both minigenome RNAs were flanked by non- coding sequences of segment 8 of FIuA and FIuB, respectively. The transfection mixture also contained a plasmid constitutively expressing Renilla luciferase, which served to normalize variation in transfection efficiency. The reporter activity was determined 24h post transfection and normalized using the Dual- Clu® Lufierase Assay System (Promega). The activity observed with transfection reactions containing Flag-CFP were set to 1 00%.
Peptide synthesis: The solid-phase synthesis of the peptides was carried out on a Pioneer automatic peptide synthesizer (Applied Biosystems, Foster City, USA) em ploying Fmoc chemistry with
TBTU/diisopropylethyl amine activation. Side chain protections were as follows: Asp, CIu, Ser, Thr and
Tyr: t-Bu; Asn, CIn and His: Trt; Arg: Pbf; Lys and Trp: Boc. Coupling time was 1 h. Double couplings were carried out if a difficult coupling was expected according to the program Peptide Companion (Coshi-
Soft/PeptiSearch, Tucson, USA). All peptides were generated as carboxyl amides by synthesis on Rapp S RAM resin (Rapp Polymere, Tubingen, Germany). Biotin was incorporated at the C-terminus of indicated peptides with Fmoc- Lys(Biotin)-OH (NovaBiochem/Merck, Nottingham, UK) and
TBTU/diisopropylethylamine activation for 1 8h, followed by coupling of Fmoc-β-Ala-OH for 1 h. Peptides were cleaved from the resin and deprotected by a 3h treatment with TFA containing 3% triisobutylsi- lane and 2% water (1 0ml/g resin). After precipitation with t-butylmethylether, the resulting crude pep- tides were purified by preparative HPLC (RP-1 8) with water/acetonitrile gradients containing 0.1 % TFA and characterized by analytical HPLC and MALDI-MS. Some peptides were synthesized by pep- tides&elephants (Nuthetal, Germany) and subsequently purified and characterized as described above.
lmmunoprecipitation experiments: H EK293T cells were transfected with the indicated plasmids in 6- well plates using Metafectene (Biontex, Martinsried, Germany). Cells were incubated 24h post transfec- tion with lysis buffer (2OmM Tris pH7.5, 1 0OmM NaCI, 0.5mM EDTA, 0.5% NP40, 1 % Protease inhibitor Mix G, (Serva, Heidelberg, Germany), I mM DTT) for 1 5 min on ice. After centrifugation by 1 3.000 rpm at 4°C supernatant was incubated with anti HA-specific antibodies coupled to agarose beads (Sigma) for 1 h at 4°C. After three washes with 1 ml of washing buffer (lysis buffer without protease inhibitor mix), bound material was eluted under denaturing conditions and separated on SDSPAGE gels and transferred to PVDF membranes. Viral polymerase subunits and GFP fusion proteins were detected with antibodies directed against the HA-tag (Covance, Berkeley, California) or H is-tag (Qiagen) or CFP- tag (Santa Cruz Biotechnology).
Plaque reduction assay: The experiments were carried out as described by Schmidke (2001 ) with modifications. Confluent MDCK cells were infected with I OOPFU of A/WSN/33, B/Yamagata/73, A/KAIM- 1 , or VSV/lndiana in PBS containing BSA at room temperature. After removal of the inoculum, cells were overlaid with medium (DMEM with 2OmM Hepes, 0.01 % DEAE Dextran, 0.001 % NaHC03) containing 1 % Oxoidagar and candidate peptides or small molecule compounds at the indicated concentrations. After incubation for 24h (VSV), 48h (A/WSN/33, A/KAN-1 ) at 37 °C with 5% CO2, or 72h at 33°C with 5% CO2 (B/Yamagata/73) respectively, cells were fixed with formaldehyde and stained with crystal violet. Plaques were counted and mean plaque number of the water control was set to 1 00%.
Enzyme-Linked Immunosorbent Assay (ELISA): For the ELISA microwell plates (Pierce) were incubated with saturating concentrations of peptides at room temperature, washed and subsequently incubated at room temperature with HA-tagged PA. To obtain PA-HA, 293T cells were seeded into 94mm-dishes, transfected with the respective plasmid and treated with lysis buffer 24h post transfection as described in detail by Mayer et al. (2007). After washing the microwell plates, the wells were incubated with an HA-specific primary antibody (Covance), followed by three washes and an incubation with a peroxidase- coupled secondary antibody (Jackson l mmuno Research, Newmarket, UK) for further 30min. After the final wash step, ABTS-substrate (Sigma, ready-to-use solution) was added and the optical density was determined at 405nm.
The competition ELISA was carried out as described above with the exception that the candidate peptide or small molecule competitor compound were added to wells of the plate with bound peptides prior to addition of the cell extract containing HA-tagged PA subunits.
Fluorescence Polarization (FP) Assay: The test sample includes a known binding pair of proteins or protein subunits including a fluorescent label, which can be analyzed according to a preferred embodiment of the present invention by fluorescence polarization. Here, we use the interaction of Influenza A virus polymerase subunit PBl , represented by the first 25, N-term inal amino acids, and subunit PA. The test sample is then contacted with a candidate peptide or small molecule inhibitor com pound and the resulting fluorescence polarization is determined. The ability of the compound to cause dissociation of or otherwise interfere with or prevent binding of the proteins or protein subunits is monitored by fluorescence polarization (FP). FP measurements allow for discrimination between fluorescently labeled bound and unbound proteins, peptides, subunits or fragments thereof. The FP of the fluorescently labeled first fragment rotates rapidly in solution and, therefore, has randomized photo-selected distributions, which result in the small observed FP. When the fluorescently labeled first fragment of the first subunit interacts with the fragment of the second subunit, which is typically a larger, more slowly rotating molecule, the rotation of the fluorescently labeled first fragment slows and the fluorescence polarization increases. Accordingly, disruption of the subunit interaction by a test compound provides a decrease in the fluorescence polarization, which is indicative of inhibition of the protein interactions. The FP measurements in the presence of a test compound can be compared with the FP measurements in the absence of the test compound. Comparison can be made manually by the operator or automatically by a computer, especially in high throughput assays using 384-well plates.
For protein purification influenza A virus polymerase subunit PA was cloned into a suitable expression vector with a C-terminally attached 6xHis-linker or hemagglutinine epitope (HA). Human 293T cells were transfected with the plasmid. Cell lysates were prepared 24 hours post transfection using lysis buffer (2OmM TrisHCI pH 7.5, 1 0OmM NaCI, 0.5mM EDTA, 0.5% NP-40, I mM DTT and 1 % Protase inhibitor mix) For purification from the lysate, PA subunit was bound to Ni- or anti-HA-agarose and washed with lysis buffer without protease mix. After elution with HA-peptide in 2OmM TrisHCI pH 7.5, 1 5OmM NaCI, 0.5mM EDTA, I mM DTT and 5% Glycerol, PA-protein was concentrated when necessary using Vivaspin20 5OK columns and frozen at -8O0C until further use. After thawing, the elution buffer was exchanged to low fluorescent grade reagents and any HA-peptide was removed simultaneously using 1 0-DC Bio-Gel columns.
Fluorescently labeled peptide corresponding to the 25 first N-terminal amino acids of Influenza A virus polymerase subunit PBl at 3 nM concentration was added to l OμM HA-PA in 2OmM TrisHCI pH 7.5, 1 5OmM NaCI, 0.5mM EDTA, I mM DTT, 5% Glycerol and l OOmg/ml bovine gamma globulin. The mix was distributed into black 384-well plates to a total volume of 20 μl per well and kept on ice. Test compounds solved in DMSO were added to a final concentration of 25μM. After incubation for 1 0 minutes at room temperature, plates were read using an Infinite F200 reader (Tecan). FP values of the wells containing test compounds were compared to wells without test compounds, without DMSO and with peptide only.
Sequence alignment: Alignments were performed with MUSCLE as described in Edgar (2004) using the full-length sequences provided from the public influenza virus database (https://www.ncbi.nlm.nih.gov/genomes/FLU/FLU.html).
Modelling: Manual docking of the mutated peptide into the PA(C)-PBl (N) crystal structure (He et al., 2008) and subsequent minimization was performed with Accelrys Discovery Studio.
Respiratory Syncytial Virus: Materials and Methods
Activity of compounds in reducing RSV induced cell death: H Ep-2 cells (obtained from ATCC) were seeded in 96-well plates (1 .5 x 1 04 cells per well) and grown in MEM-alpha medium containing 1 0% FBS (Cibco-BRL) for 24 h. To infect cells, 500 pfu of RSV Long strain (obtained from ATCC) were added in 50 μl of OptiMEM (Cibco-BRL) for 1 h. Cells were then incubated in the presence of a serial dilution of compounds (from 1 00 to 0.14 μM) in MEM-alpha containing 2% FBS for 72 h. Cells were fixed in 3.7% formaldehyde and stained with 0.025% of crystal violet (Sigma). The integrity of the cellular monolayer was measured at 540 nm using a microplate reader. The activity of the com pounds to reduce virus- induced cell death is expressed as the mean of three independent experiments each performed in triplicates.
Experimental Results
Protein-protein interactions (PPIs) are crucial to most, if not all, biological processes. Of the roughly 30,000 protein sequences that comprise the human proteome, only about 1 % have been successfully targeted with small-molecule drugs. Yet, most of the conventional targets in drug discovery fall into the same few structural or functional families such as enzymes or C protein-coupled receptors (CPCRs). They typically share the property that the natural substrates or ligands, with which they interact are themselves small organic molecules. Historically there has been notably little success in developing drug-like inhibitors of proteins whose natural ligands are other proteins. Designing a small molecule to bind to a protein-protein interface and inhibit the interaction poses several challenges, including the initial identification of suitable PPIs, the surface area of the interface, and the localization of "hot spots". Thus, small molecule inhibition of PPIs is a challenging area in drug discovery.
The present invention uses the fact that proteomes of many viruses and PPIs crucial for viral replication are described in the literature. For any proteome of interest, this data is according to the novel method supplemented with proteomic approaches for identification of PPIs like yeast two-hybrid or co-immuno precipitation screening in order to identify potential target regions for development of PPI inhibitors. Subsequently, a unique combination of phylogenetic analysis and structure prediction or structure analysis (where applicable) of the protein partners involved detects druggable protein-binding domains. Within the present disclosure, the term druggable denotes preferably protein-binding domains which can be blocked, altered or modified by small molecules in a way that the protein-protein interaction is inhibited or disrupted. The term small molecules denotes organic molecules, preferably synthetic organic molecules (not peptides), which have a molecular weight below 1 500, preferably below 1 000 and most preferred below 500 u. It has been found, that these domains bear a couple of characteristic features: (i) helical structure, (ii) hydrophobic character and (iii) high conservation among all virus strains. It has been shown that they tend to be located at a term inal end of the protein or are located on their surface. The peptides corresponding to these potential binding domains are synthesized in an overlapping way and tested for their ability to bind the protein partner involved in the PPI. If peptides resembling short, (less than 20 amino acids) continuous binding domains are identified, these are used for the development of a binding assay, preferably an ELISA or fluorescence polarization (FP) assay, which is afterwards employed in a high-throughput screening campaign for small molecule and/or peptidic inhibitors of the PPI.
The PPI inhibitors identified by the novel method according to the present invention, as opposed to conventional active site inhibitors, could offer a particular advantage when it comes to antivirals since it should be safe to assume that resistance development occurs at a much slower pace.
In order to identify chemical compounds that efficiently interrupt or disrupt the interaction between PBl and PA, for example by binding to the inhibition target on PA of FIuA and FIuB, the ELISA assay described above for the influenza peptides was repeated with a number of small molecule com pounds obtained from corresponding compound libraries from Maybridge Ltd., Cambridge, UK (www.maybridge.com). The tested compounds are listed in Ta ble 4, together with their systematic name, the product code, and the result for the ELISA assay.
Table 4: Compounds tested with influenza ELISA
Comp. ID Compound name Product Active in code EUSA
PKE064 1 -{4-Methyl-2-[5-(2-thιenyl)-2-thιenyl]-l ,3-thιazol-5-yl}ethan-l -one SEW04843 yes
PKE088 4-(4-nιtrophenyl)-2-[5-(2-thιenyl)-2-thιenyl]-l ,3-thιazole SEW04834 no
PKE089 4-(4-chlorophenyl)-2-[5-(2-thιenyl)-2-thιenyl]-l ,3-thιazole SEW04835 yes
PKE090 2-[5-(2-thιenyl)-2-thιenyl]-4-[4-(trιfluoromethyl)phenyl)-1 ,3-thιazole SEW04836 yes
PKE091 4-(3,4-dιfluorophenyl)-2-[5-(2-thιenyl)-2-thιenyl]-l ,3-thιazole SEW04837 no
PKE092 4-(tert-butyl)-2-[5-(2-thιenyl)-2-thιenyl]-l ,3-thιazole SEW04838 yes
PKE093 (2-thιenyl)(5-{2-[5-(2-thιenyl)-2-thιenyl]-4-methyl-l ,3-thιazol-5-yl}-l H- SEW04867 yes pyrazol-1 -yl)methanone
PKE094 1 -(5-{2-[5-(2-thιenyl)-2-thιenyl]-4-methyl-l ,3-thιazol-5-yl}-l H-pyrazol-1 - SEW04868 yes yl)-2,2-dιmethylpropan-l -one
PKE095 (5-{2-[5-(2-thιenyl)-2-thιenyl]-4-methyl-l ,3-thιazol-5-yl}-l H-pyrazol-1 - SEW04870 yes yl)[3-(trιfluoromethyl)phenyl]methanone
PKE096 5-[5-(4-benzo[b]furan-2-yl-l ,3-thιazol-2-yl)thιen-2-yl]ιsoxazole HTS02299 yes
PKE097 methyl [2-(5-ιsoxazol-5-ylthιen-2-yl)-5-methyl-l ,3-thιazol-4-yl]acetate HTS02301 yes
PKE098 2-[5-(4-phenyl-l ,3-thιazol-2-yl)-2-thιenyl] pyridine HTS03053 yes
PKE099 2-{5-[4-(tert-butyl)-l ,3-thιazol-2-yl]-2-thιenyl}pyrιdιne HTS03331 yes
PKEl OO ethyl 2-(5-ιsoxazol-5-ylthιen-2-yl)-4-methyl-l ,3-thιazole-5-carboxylate HTSl 0580 yes
PKEl Ol 4-methyl-2-[4-phenyl-5-(trιfluoromethyl)-2-thιenyl]-l ,3-thιazole-5- HTSl 0582 yes carboxyhc acid
PKE141 N-{[2-(2-thιenyl)-l ,3-thιazol-4-yl]methyl} benzenesulfonamide SCROl 1 95 no
PKEl 42 N-{[2-(2-thιenyl)-l ,3-thιazol-4-yl]methyl} benzamide SCROl 314 no
PKE143 1 -[4-methyl-2-(2-thιenyl)-l ,3-thιazol-5-yl]ethan-l -one SPB02598 no
PKEl 44 4-methyl-5-(l H-pyrazol-5-yl)-2-(2-thιenyl)-l ,3-thιazole SPB0261 2 no
PKE145 4-methyl-5-[2-(methylthιo)pyrιmιdιn4-yl]-2-(2-thιenyl)-l ,3-thιazole SPB0261 8 no
PKEl 46 cyclopropyl{5-[4-methyl-2-(2-thιenyl)-l ,3-thιazol-5-yl]-l H-pyrazol-1 - SPB02622 no yljmethanone
PKEl 47 N 1 -ιsopropyl-5-[4-methyl-2-(2-thιenyl)-l ,3-thιazol-5-yl]-l H-pyrazole-1 - SPB02625 no carboxamide
PKEl 66 l -methyl-N-{[2-(2-thιenyl)-l ,3-thιazol-4-yl]methyl}-l H-ιmιdazole4- SCROl 202 no sulfonamide
PKEl 67 N-{[2-(2-thιenyl)-l ,3-thιazol-4-yl] methyl} cyclopropanecarboxamide SCROl 296 no
PKEl 68 4-methyl-2-(2-thιenyl)-l ,3-thιazole-5-carbohydrazιde SPB02247 no For some of the compounds with positive ELISA prescreening the inhibitory concentrations (IC50) have been determined in a plaque reduction assay as described above for the influenza peptide studies. For another group of compounds a competitive ELISA assay as described above for the influenza peptide studies has been carried out to determine the maximum ELISA inhibition at the highest concentration used (1 000 μM). The results are shown in Table 5.
Table 5: Influenza inhibitory concentrations (IC5o) of compounds
Compound ID IC50 [μM] Max. Inhibition (Plaque Reduct.) (ELISA) at 1000 μM
PKE064 002 45%
PKE088 n d 20%
PKE089 n d 30%
PKE090 n d 37%
PKE091 n d 20%
PKE092 20
PKE093 n d 36%
PKE094 009
PKE095 n d 36%
PKE096 n d 30%
PKE097 n d 30%
PKE098 20
PKE099 40
PKEl OO n d 28%
PKEl Ol n d 34%
PKEl 43 n d inactive
The compounds that have been found so far to be effective in binding to PA (Table 4, 5) have a basic structure of 2-(thien-2-yl)-thiazole. Comparing compound PKE 92 and PKE 99 one may assume that the use of 2-pyridyl as terminal residue slightly increases the inhibition activity of a compound according to the invention. The removal of the terminal thienyl residue, however, completely destroys the activity of a com pound, as can be seen for example when comparing PKE 64 and PKE 143.
Actually none of the tested 2-(thien-2-yl)-thiazole compounds in Table 5 without an aromatic terminal residue such as 2-thienyl, 2-pyridyl, 5-isoxazolyl, or phenyl at its thienyl ring position has been found active. Without being bound to any theory, it is assumed that such a residue plays a crucial role in the binding ability of a compound according to the invention.
A similar screening was carried out with the plaque reduction assay as described above, with influenza virus (A/WSN/33) and in addition also with RSV (Long strain), for the above-mentioned com pounds and with a number of newly synthesized compounds. Said novel compounds are defined by their structural formula in Figure 5. The synthesis of the compounds is disclosed in the synthesis section of the description. The results of the screening are given in Table 6, with the maximum inhibition obtained and, if determinable, the IC50 value. If in an influenza pre-screening assay (competitive ELISA or other) the compound was found to be inactive or having a too high ELISA IC50, the influenza assay was not carried out for efficiency reasons.
Table 6: Influenza and RSV inhibition of compounds
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000024_0001
Figure imgf000025_0001
* The compounds PKE 064 and PKE 296 are identical. However, PKE 064 was obtained from a commercial compound library with restricted degree of purity and a considerable shelf time, while PKE 296 is the freshly synthesized, pure compound (>99%). It is assumed that the difference in the activities of PKE 064 and PKE 296 are due to impurities in PKE 064.
The assessed class of compounds seems to be effective in the inhibition of replication of certain virus types. A number of compounds very effectively inhibited the replication of influenza virus, particularly influenza A. Surprisingly it was found that a number of other compounds are also effective in the inhibition of RSV replication, and certain compounds inhibit the replication of both virus types.
Without being bound to any theory, it seems that the compounds according to the invention can be very effective broad band inhibitors of virus replication, and thus are a valuable source of effective new medicaments against certain types of the orthomyxoviridae and paramyxoviridae families. Synthesis of compounds
The synthesis of the compounds according to the invention is described in the following chemical description part.
General reaction scheme for ester, acid, amide and alcohol thiazole derivatives
Figure imgf000026_0001
άCSiϊ ΪSSfS&S "SS"!' iβOδSϊ
Figure imgf000026_0002
Figure imgf000026_0003
In this general reaction scheme, Ar1 is either a thiophene, a substituted thiophene, a furan or a substituted furan. M can be a boronic acid, a boronic ester, or a stannyl functional group. RI can be H, CH3, te/t-butyl, or CF3 group. Starting 2-bromo-thiazole ester a is directly available or can be prepared accord- i o ing to standard procedure described in the litterature. For example, in the case of the te/t-butyl series (RI = tBu), the synthesis of a follows the procedures described in J. Chem. Soc, Perkin Trans /, 1 982, 1 52-1 64, or, for other cases, a can be prepared by a simple Sandmeyer reaction from the corresponding 2-aminothiazole ester.
Ester compounds (I): Compounds of formula (I), in the ester series, are generally prepared according to
15 the reaction scheme shown above, from a via b, c to (I). A classic palladium (Pd) catalyzed coupling reaction, such as a Suzuki coupling reaction or a Stille coupling reaction leads to bicyclic ester b. Follows a classic procedure of Suzuki reaction, using Pd catalyst like tetrakis palladium triphenylphosphine, a base, like aqueous NaHCO3 or cesium fluoride, in a classic organic solvent such as N1N- dimethylformamide or 1 ,2-Dimethoxyethane. Brominated compound c is achieved by direct bromina- tion, using Br2 in acetic acid media, /V-bromosuccinimide in a usual solvent, or copper (II) bromide in the presence of a nitrite in acetonitrile. Then another classic Pd catalyzed coupling reaction, a Suzuki reaction or a Stille reaction, yields the expected formula (I) compounds.
In some particular cases, bicyclic systems Ar2-ArI-M are directly available, as for thienyl-thienyl-M. Then, compounds of formula (I) are prepared from compound a by a one pot Suzuki or Stille coupling reaction, using classic Pd catalyst, such as tetrakis palladium triphenylphosphine, in the presence of a base like NaHCOs or cesium fluoride, in a usual solvent like /V,/V-dimethylformamide or 1 ,2-dimethoxyethane.
Acid compounds (II): Hydrolysis of the ester functional group of compounds (I) to the corresponding carboxylic acid (II) is made with lithium hydroxide in a methanol/water solution or in a tetra hyd rofu- ran/water solution. All acid compounds can be prepared as sodium salts or ammonium salts by usual procedures.
Alcohol compounds (III): Reduction of the ester functional group of compounds (I) to the corresponding alcohol is achieved in the presence of lithium aluminum hydride in a classic solvent such as tetrahy- drofuran, giving the compounds of formula (III).
Amide compounds (IV): The ester function of the brominated compound c is hydrolyzed with lithium hydroxyde to the corresponding carboxylic acid d. Then, the formation of the amid compound e can be achieved by classical reactions, such as formation of an acid chloride followed by addition of an amine, in the presence of a strong base like triethylamine, in a classical solvent like acetone or dichloro- methane. A Pd catalyzed coupling reaction like a Suzuki reaction or a Stille reaction gives the final compounds of formula (IV).
Acetyl compounds (V)
Figure imgf000027_0001
Figure imgf000027_0002
The preparation of acetyl compounds of formula (V) follows the reaction scheme shown above. The 2- aminothiazole f is brominated via a Sandmeyer reaction (EP 1 894930, 2008, or Organic Letters, 2003, 7963, or J. Chem. Soc Perkin Trans. J , 1 982, 1 59), using copper (II) bromide or NaBr/CuS04, in the presence of a nitrite, either isoamylnitrite or sodium nitrite, in a solvent like acetonitrile. Then, the same synthesis steps as for formula (I) compounds apply: Pd catalyzed coupling reaction, bromination, and final Pd catalyzed coupling reaction lead to com pounds (V).
General reaction scheme for compounds with a modified central thiophene (ArI not being thio- phene)
Figure imgf000028_0001
(H) (I) esier series
I n the case where Ar1 is a phenyl group or a pyridine, the syntheses of ester compounds (I) and acid compounds (II) follow the reaction scheme shown above. M can be a boronic acid, a boronic ester, or a stannyl functional group. Rl can be H, CH3, tert-butyl, or CF3 group. Starting 2-bromothiazole ester a undergoes a Pd catalyzed coupling reaction, like a Suzuki or a Stille reaction, to yield the alcohol j. This alcohol is activated by use of triflic anhydride in the presence of 2,6-lutidine and DMAP in a solvent like dichloromethane (J. Chem. Soc, Chem. Comm., 1 987, 904-905). Then another Pd catalyzed coupling reaction such as a Suzuki reaction, leads to the formula (I) ester compounds. A classic hydrolysis with lithium hydroxide affords the corresponding acid compounds (II).
Figure imgf000028_0002
(I) aster series
Figure imgf000028_0003
(II) acid series For some compounds where Ar1 is a pyridine for example, the above-shown reaction scheme is used to prepare formula (I) and formula (II) compounds. Reaction of O.O'-diethyl dithiophosphonate and starting bromonitrile I in a pyridine/water solution leads to thioamide m (Phosphorus and Sulfur and related Elements, 1 985, 297-306). Thiazole formation is achieved by action of ethyl-2-chloroacetate in a polar solvent like ethanol. Then a usual Pd catalyzed coupling reaction affords the formula (I) ester compounds, and a following hydrolysis leads to the acid compounds (II).
General reaction scheme for compounds with a central amide group
BDrI
Figure imgf000029_0001
Figure imgf000029_0002
n (Vl) (VII)
Figure imgf000029_0003
(IX) (VIII)
In this series, Ar1 is a thiophene, X can be H or Br. Amides of formula (Vl) are prepared according to classic reaction, like peptide coupling reaction (J. Am. Chem. Soc, 2000, 6382-6394) using HOBt and EDCI in a usual solvent such as /V,/V-dimethylformamide. When X = Br, a Pd catalyzed coupling reaction, like a Suzuki or a Stille reaction leads to compounds of formula (VII). Methylation of the nitrogen is achieved by using sodium hydride and methyl iodide in /V,/V-dimethylformamide, affording compounds of formula (VIII). Then a classic hydrolysis of the ester function leads to compounds of formula (IX).
General reaction scheme for compounds with other thiazole substitution:
Figure imgf000029_0004
Ar1 , Ar2 = substituted (or not) thiophene, phenyl, pyridine, Ar s R9 benzothiophene, Ar2-^ 1 -^ V"
R1 ,R2 = H, Me, COOEt, COOH, N R1
CONHOH, substituted (or not) tetrazole, oxazole,
(Vl)
In this series, ArI is a thiophene, and either Rl or R2 or both can be a H, CH3, ester, acid, ester isoster or acid isoster (tetrazole, isoxazole hydroxamic acid or derivative). The synthesis of compounds of formula (Vl) follows the reaction scheme shown above. 2-Aminothiazole o is brominated via a Sandmeyer reaction. Then, the same synthesis steps as for formula (I) compounds apply: Pd catalyzed coupling reaction, bromination, and final Pd catalyzed coupling reaction lead to com pounds (Vl).
General reaction scheme for compounds with a central carbonyl group:
Figure imgf000030_0001
s b (VII)
In one particular case (Example D l -I , ArI = Ar2 = thiophene), we prepared a compound with a central carbonyl functional group.
The synthesis of such compounds of formula (VII) follows the reaction scheme shown above. Acid chlo- ride s is coupled with biaryl b in the presence of a Lewis Acid such as FeCI3, AICI3, TiCI4,etc. Direct coupling reaction leads to compounds (VII).
Synthesis for thiazole replacement:
Figure imgf000030_0002
(X) (Xl) (XII) (XIII)
R1 ,R2,R4 = H, Me, CF3, COOEt, COOH, CHO,
In some specific cases where ArI is a thiophene and Ar2 is a thiophene or a benzo[b]thiophene, term inal 2-linked-l ,3-thiazole has been replaced by other substituted cycles, such as thiophene, 5-lin ked-1 ,3- thiazole, 1 ,3,4-thiadiazole, or substituted phenyl. The scheme shown above summarizes these structures. The syntheses of compounds (X), (Xl), (XII) and (XIII) follow the same synthetic pathway described in reaction scheme 2 for thiazole derivatives.
Specific Examples
5-acetyl-l ,3-thiazole series
Example Al -1 -1 : l -(2-bromo-4-methyl-l ,3-thiazol-5-yl)-ethanone: To a mixture of 5-acetyl-2-amino-4- methyl-thiazole (4.50 g, 28.9 mmol) and anhydrous copper (II) bromide (6.46 g, 28.9 mmol) in dry ace- tonitrile (1 20 ml) was added isoamyl nitrite (1 1 .6 ml, 86.3 mmol). The mixture was stirred at 600C for 4 hours and at room temperature for 1 6 hours. The solvent was evaporated in vacuo and the residue was purified by silica gel chromatography (EtOAc-cyclohexane: 1/1 ) to afford a yellow oil (67%). 1 H NMR (CDCI3) δ: 2.71 (s, 3H), 2.50 (s, 3H). MS (ESI+) m/z: 21 9.3-221 .3 [M + H]+.
Example Al -1 -2 / PKE 143: l -(4-methyl-2-thiophen-2-yl-thiazol-5-yl)-ethanone: To a mixture of thio- phene-2-boronic acid (3.37 g, 26.3 mmol), l -(2-bromo4-methyl-thiazol-5-yl)-ethanone Al -1 -1 (4.14 g, 1 8.8 mmol), and 2M aqueous Na2CO3 (20 ml, 40.0 mmol ) in 1 30 ml of a DME-H2O solution (9/1 ) degassed under argon, was added Pd(PPh3J4 (1 .09 g, 0.94 mmol). The reaction mixture was degassed under argon and heated up to 900C for 1 6 h. The reaction mixture was quenched by addition of water, diluted with EtOAc, filtered over celite and extracted with EtOAc. The organic layer was washed twice with water, dried over MgSO4 and concentrated in vacuo. Chromatography of the residue on silica gel (cyclohexane-EtOAc: 8/2) afforded a yellow solid (83%). 1 H NMR (CDCI3) δ: 7.61 (dd, J = 3.8, 1 .1 Hz, 1 H), 7.48 (dd, J = 5.1 , 1 .1 Hz, 1 H), 7.1 1 (dd, J = 5.1 , 3.8 Hz, 1 H), 2.75 (s, 3H), 2.55 (s, 3H). MS (ESI+) m/z: 224.0 [M + H]+.
Example Al -1 -3: l -[2-(5-Bromo-thiophen-2-yl)-4-methyl-l ,3-thiazol-5-yl]-ethanone: To a mixture of l -(4- methyl-2-thiophen-2-yl-thiazol-5-yl)-ethanone Al -1 -2 (3.25 g, 14.5 mmol) and anhydrous copper (II) bromide (3.25 g, 14.5 mmol) in dry acetonitrile (1 00 ml) was added isoamyl nitrite (1 .70 ml, 1 2.6 mmol). The mixture was stirred at 600C for 20 hours. The solvent was evaporated in vacuo and the residue was purified by silica gel chromatography (EtOAc-cyclohexane: 2/8) to afford a yellow solid
(80%). λ H NMR (CDCI3) δ: 7.34 (d, J = 3.9 Hz, 1 H), 7.07 (d, J = 3.9 Hz, 1 H), 1 .73 (s, 3H), 2.55 (s, 3H). MS (ESI+) m/z: 301 .8-303.8 [M + H]+.
Example Al -2 / PKE 296: l -(2-[2,2']Bithiophen-5-yl4-methyl-thiazol-5-yl)-ethanone: To a mixture of thiophene-2-boronic acid (0.1 2 g, 0.92 mmol), l -[2-(5-Bromo-thiophen-2-yl)-4-methyl-thiazol-5-yl]- ethanone Al -1 -3 (0.20 g, 0.66 mmol), and 2M aqueous Na2CO3 (0.66 ml, 1 .22 mmol) in 1 0 ml of a DME-H2O (9/1 ) solution degassed under argon was added Pd(PPh3J4 (38 mg, 33 mol). The reaction mixture was degassed under argon and heated up to reflux for 1 8 h. The reaction mixture was quenched by addition of water, diluted with EtOAc, filtered over celite and extracted with EtOAc. The organic layer was washed twice with water, dried over MgSO4 and concentrated in vacuo. Chromatography of the residue on silica gel (cyclohexane-EtOAc: 80/20) afforded a yellow solid (83%). 1 H NMR (DMSO) δ: 7.78 (d, J = 4.0 Hz, 1 H), 7.63 (dd, J = 5.1 , 1 .1 Hz, 1 H), 7.50 (dd, J = 3.6, 1 .1 Hz, 1 H), 7.39 (d, J = 4.0 Hz, 1 H), 7.1 5 (dd, J = 5.1 , 3.6 Hz, 1 H), 2.66 (s, 3H), 2.55 (s, 3H). MS (ESI+) m/z: 306.0 [M + H]+. Mp: 1 38°C-140°C.
Example Al -3 / PKE 321 : l -[4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazol-5-yl]-ethanone: Method as for Al -2, using pyridine-3-boronic acid; yellow solid (56%). λ H NMR (DMSO) δ: 8.99 (d, J = 0.8 Hz, 1 H), 8.55 (dd, J = 4.7, 1 .5 Hz, 1 H), 8.1 7-8.1 3 (m, 1 H), 7.88 (d, J = 3.9 Hz, 1 H), 7.75 (d, J = 4.1 Hz, 1 H), 7.48 (dd, J = 7.9, 4.7 Hz, 1 H), 2.66 (s, 3H), 2.55 (s, 3H). MS (ESI+) m/z: 301 .0 [M + H]+. Mp: 1 72 °C- 1 75°C.
Example Al -4 / PKE 329: l -{2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazol-5-yl]-ethanone: Method as for Al -2, using 4-methoxy-phenyl boronic acid; yellow solid (35%). 1 H NMR (DMSO) δ: 7.81 (d, J = 4.0 Hz, 1 H), 7.70 (d, J = 8.8 Hz, 2H), 7.51 (d, J = 4.0 Hz, 1 H), 7.02, (d, J = 8.8 Hz, 2H), 3.80 (s, 3H), 2.66 (s, 3H), 2.56 (s, 3H). MS (ESI+) m/z: 330.2 [M + H]+. Mp: 1 52 °C-1 55°C.
Example Al -5 / PKE 346: l -[2-(5'-acetyl-[2,2']bithiophenyl-5-yl)-4-methyl-thiazol-5-yl]-ethanone: Method as for Al -2, using 2-acetyl-thiophene-5-boronic acid; yellow solid (22%). 1 H NMR (DMSO) δ: 7.95 (d, J = 3.9 Hz, 1 H), 7.86 (d, J = 4.1 Hz, 1 H), 7.62 (d, J = 3.9 Hz, 1 H), 7.61 (d, J = 4.0 Hz, 1 H), 2.67 (s, 3H), 2.57 (s, 3H), 2.55 (s, 3H). MS (ESI+) m/z: 348.1 [M + H]+. Mp: 1 64°C-1 66°C.
Example Al -6 / PKE 352: l -{2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazol-5-yl}- ethanone: Method as for Al -2, using 2-trifluoromethoxy-phenyl boronic acid; yellow solid (91 %). 1 H NMR (DMSO) δ: 7.96-7.93 (m, I H), 7.87 (d, J = 3.9 Hz, I H), 7.65 (d, J = 3.9 Hz, I H), 7.55-7.49 (m, 3H), 2.67 (s, 3H), 2.56 (s, 3H). MS (ESI+) m/z: 384.1 [M + H]+. Mp: 1 01 0C-1 030C.
Example Al -7 / PKE 350: l -[2-(5-Benzo[b]thiophen-2-yl)-4-methyl-thiazol-5-yl]-ethanone: Method as for Al -2, using benzo[b]thiophene-2-boronic acid; yellow solid (66%). 1 H NMR (DMSO) δ: 8.00-7.97 (m, 1 H), 7.88-7.84 (m, 3H), 7.54 (d, J = 4.0 Hz, 1 H), 7.43-7.39 (m, 2H), 2.68 (s, 3H), 2.56 (s, 3H). MS (ESI+) m/z: 356.2 [M + H]+. Mp: 1 69°C-1 71 °C.
Example Al -8 / PKE 385: l -[4-Methyl-2-(5'-methyl-[2,2']bithiophenyl-5-yl)-thiazol-5-yl]-ethanone: Method as for Al -2, using 5-methyl-thiophene-2-boronic acid; yellow solid (14%). 1 H NMR (CDCI3) δ: 7.50 (d, J = 4.2 Hz, 1 H), 7.07 (m, 2H), 6.70 (d, J = 3.2 Hz, 1 H), 2.75 (s, 3H), 2.55 (s, 3H), 2.50 (s, 3H). MS (ESI+) m/z: 320.1 [M + H]+. Mp: 1 27 °C-1 300C.
Example Al -9 / PKE 329: l -[4-Methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazol-5-yl]-ethanone hydrochloride salt (hydrochloride of compound PKE 321 ): Method as for Bl -60, using l -[4-methyl-2-(5-pyridin-3-yl- thiophen-2-yl)-thiazol-5-yl]-ethanone Al -3 / PKE 321 ; yellow solid (89%). 1 H NMR (DMSO) δ: 9.1 9 (d, J = 5.1 Hz, I H), 8.71 (dd, J = 5.1 , 1 .3 Hz, I H), 8.51 (dt, J = 8.1 , 1 .3 Hz, I H), 7.95 (d, J = 4.1 Hz, I H), 7.89 (d, J = 4.0 Hz, 1 H), 7.79 (dd, J = 8.3, 5.3 Hz, 1 H), 2.68 (s, 3H), 2.58 (s, 3H). Mp: 252 °C-255°C.
4-(tert-butyl)-l ,3-thiazole series
Example A2-1 -1 : 2-Bromo-4-tøt-butyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -1 , using 2-amino-4-tert-butyl-thiazole-5-carboxylic acid ethyl ester (prepared according to Journal of the Chemical Society, Perkin Trans I, 1 982, 1 52-1 64); white solid (94%). 1 H NMR (CDCI3) δ: 4.30 (q, J = 7.1 Hz, 2H), 1 .46 (s, 9H), 1 .35 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 291 .9-293.5 [M + H]+.
Example A2-1 -2: 4-tøt-Butyl-2-thiophen-2-yl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al - 1 -2, using 2-bromo4-tert-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -1 ; beige solid (96%). 1 H NMR (CDCI3) δ: 7.54 (dd, J = 3.7, 1 .1 Hz, 1 H), 7.42 (dd, J = 5.1 , 1 .1 Hz, 1 H), 7.08 (dd, J = 5.1 , 3.7 Hz, 1 H), 4.32 (q, J = 7.1 Hz, 2H), 1 .52 (s, 9H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 296.0 [M + H]+.
Example A2-1 -3: 2-(5-Bromo-thiophen-2-yl)-4-tert-butyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -2, using 4-tert-butyl-2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester A2-1 -2; yellow solid (98%). 1 H NMR (DMSO) δ: 7.65 (d, J = 4.0 Hz, I H), 7.33 (d, J = 4.0 Hz, I H), 4.27 (q, J = 7.1 Hz, 2H), 1 .42 (s, 9H), 1 .27 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 373.9-375.9 [M + H]+.
Example A2-M / PKE 259: 2-[2,2 ']Bithiophenyl-5-yl-4-tøt-butyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-thiophen-2-yl)4-te/t-butyl-thiazole-5-carboxylic acid ethyl ester A2- 1 -3; yellow solid (75%). 1 H NMR (CDCI3) δ: 7.43 (d, J= 3.9 Hz, 1 H), 7.29-7.26 (m, 2H), 7.14 (d, J = 3.9 Hz, 1 H), 7.05 (dd, J = 4.9, 3.8 Hz, 1 H), 4.32 (q, J = 7.1 Hz, 2H), 1 .52 (s, 9H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 378.2 [M + H]+. Mp: 1 1 1 0C-1 1 4°C.
Example A2-2 / PKE 357: 2-[2,2 ']Bithiophenyl-5-yl-4-tert-butyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[2,2 ']bithiophenyl-5-yl-4-tøt-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -4; yellow solid (82%). ] H NMR (DMSO) δ: 1 3.37 (br s, 1 H), 7.69 (d, J = 3.9 Hz, 1 H), 7.59 (d, J = 5.0 Hz, 1 H), 7.48 (d, J = 2.6 Hz, I H), 7.35 (d, J = 3.8 Hz, I H), 7.1 2 (dd, J = 4.9, 3.7 Hz, I H), 1 .45 (s, 9H). MS (ESI+) m/z: 350.1 [M + H]+. Mp: 205°C-207 °C.
Example A2-3 / PKE 293: 4-te/t-Butyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzene boronic acid and 2-(5-bromo-thiophen-2-yl)4-tert-butyl-thiazole-5- carboxylic acid ethyl ester A2-1 -3; yellow solid (91 %). 1 H NMR (CDCI3) δ: 7.81 (d, J = 4.0 Hz, I H), 7.75 (td, J = 7.0, 1 .4 Hz, 2H), 7.60 (d, J = 4.0 Hz, 1 H), 7.44 (tt, J = 7.2, 1 .5 Hz, 2H), 7.36 (tt, J = 7.2, 1 .3 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 1 .46 (s, 9H), 1 .29 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 372.2 [M + H]+. Mp: 1 29°C-1 32 °C.
Example KlA / PKE 367: 4-tert-Butyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-te/t-butyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A2-3; pale yellow solid (77%). 1 H NMR (DMSO) δ: 1 3.40 (br s, 1 H), 7.76-7.73 (m, 3H), 7.59 (d, J = 3.9 Hz, 1 H), 7.46-7.35 (m, 3H), 1 .46 (s, 9H). MS (ESI+) m/z: 344.1 [M + H]+. Mp: 21 2 °C-21 4°C. Example A2-5 / PKE 316: 4-ferf-Butyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using pyridine-3-boronic acid and 2-(5-bromo-thiophen-2-yl)4-tøt-butyl- thiazole-5-carboxylic acid ethyl ester A2-l -3; yellow solid (66%). 1 H NMR (DMSO) δ: 8.98 (d, J = 1 .8 Hz, I H), 8.54 (dd, J = 4.8, 1 .7 Hz, I H), 8.14 (ddd, J = 8.0, 2.4, 1 .6 Hz, I H), 7.85 (d, J = 4.0 Hz, I H), 7.72 (d, J = 4.0 Hz, 1 H), 7.46 (ddd, J = 8.0, 4.8, 0.7 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 1 .46 (s, 9H), 1 .29 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 373.2 [M + H]+. Mp: 1 1 00C-1 1 3°C.
Example A2-6 / PKE 320: 4-teΛf-Butyl-2-[5-(4-cyano-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-cyanobenzene boronic acid and 2-(5-bromo-thiophen-2-yl)-4-tert- butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (66%). 1 H NMR (DMSO) δ: 7.95 (dd, J = 6.7, 1 .9 Hz, 2H), 7.87 (dd, J = 8.0, 2.0 Hz, 2H), 7.85 (d, J = 4.0 Hz, 1 H), 7.79 (d, J = 4.0 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 1 .45 (s, 9H), 1 .28 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 397.2 [M + H]+. Mp: 1 65°C- 1 67 °C.
Example A2-7 / PKE 331 : 4-teΛf-Butyl-2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylic acid ethyl ester: Method as for Al -2, using 4-trifluoromethoxy-phenyl boronic acid and 2-(5- bromo-thiophen-2-yl)4-tert"-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (92%). 1 H NMR (DMSO) δ: 7.88 (td, J = 8.7, 2.1 Hz, 2H), 7.82 (d, J = 4.0 Hz, 1 H), 7.64 (d, J = 4.0 Hz, 1 H), 7.42 (d, J = 8.0 Hz, 2H), 4.28 (q, J = 7.1 Hz, 2H), 1 .45 (s, 9H), 1 .29 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 456.2 [M + H]+. Mp: 69°C-72 °C.
Example A2-8 / PKE 369: 4-teΛf-Butyl-2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-teΛf-butyl-2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]- thiazole-5-carboxylic acid ethyl ester A2-7; yellow solid (27%). 1 H NMR (DMSO) δ: 1 3.47 (br s, I H), 7.88 (d, J = 8.8 Hz, 2H), 7.78 (d, J = 4.0 Hz, 1 H), 7.64 (d, J = 4.0 Hz, 1 H), 7.43 (d, J = 8.1 Hz, 2H), 1 .45 (s, 9H). MS (ESI+) m/z: 428.2 [M + H]+. Mp: 206°C-209°C.
Example A2-9 / PKE 332: 4-teΛf-Butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 3-methoxy-phenyl boronic acid and 2-(5-bromo-thiophen-2-yl)-4- te/t-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (60%). 1 H NMR (DMSO) δ: 7.81 (d, J = 4.0 Hz, 1 H), 7.63 (d, J = 4.0 Hz, 1 H), 7.36 (d, J = 7.7 Hz, 1 H), 7.33-7.26 (m, 2H), 6.94 (ddd, J = 7.6, 2.5, 1 .5 Hz, 1 H), 4.27 (q, J = 7.1 Hz, 2H), 3.81 (s, 3H), 1 .45 (s, 9H), 1 .28 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 402.2 [M + H]+. Mp: 1 01 0C-1 040C.
Example A2-1 0 / PKE 344: 4-teΛf-Butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-teΛf-Butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester A2-9; yellow solid (95%). 1 H NMR (DMSO) δ: 1 3.42 (br s, 1 H), 7.74 (d, J = 4.0 Hz, I H), 7.61 (d, J = 4.0 Hz, I H), 7.35-7.25 (m, 3H), 6.94 (d, J = 7.5 Hz, I H), 3.81 (s, 3H), 1 .46 (s, 9H). MS (ESI+) m/z: 374.2 [M + H]+. Mp: 1 86°C-1 89°C.
Example A2-1 1 / PKE 354: 4-te^Butyl-2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methoxy-phenyl boronic acid and 2-(5-bromo-thiophen-2-yl)-4- te/t-butyl-thiazole-S-carboxylic acid ethyl ester A2-1 -3; green solid (79%). 1 H NMR (DMSO) δ: 7.75 (d, J = 4.0 Hz, 1 H), 7.68 (d, J = 8.9 Hz, 2H), 7.46 (d, J = 4.0 Hz, 1 H), 6.99 (d, J = 8.9 Hz, 2H), 4.27 (q, J = 7.1 Hz, 2H), 3.78 (s, 3H), 1 .45 (s, 9H), 1 .28 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 402.3 [M + H]+. Mp: 87 °C-90°C.
Example A2-1 2 / PKE 355: 4-teΛf-Butyl-2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-teΛf-Butyl-2-[5-(4-methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester A2-1 1 ; yellow solid (81 %). 1 H NMR (DMSO) δ: 1 3.44 (br s, I H), 7.69 (d, J= 2.0 Hz, I H), 7.68 (d, J = 8.7 Hz, 2H), 7.45 (d, J = 3.9 Hz, 1 H), 6.99 (d, J = 8.9 Hz, 2H), 3.78 (s, 3H), 1 .45 (s, 9H). MS (ESI+) m/z: 374.2 [M + H]+. Mp: 1 95°C-1 98°C.
Example A2-1 3 / PKE 358: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)4-tøt-butyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen- 2-yl)-4-tøt-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (83%). 1 H NMR (CDCI3) δ: 7.81 -7.73 (m, 2 H), 7.50 (s, 1 H), 7.47 (d, J = 3.9 Hz, 1 H), 7.37-7.30 (m, 2H), 7.25 (d, J = 3.8 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 1 .53 (s, 9H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 428.2 [M + H]+. Mp: 1 68°C-1 71 °C.
Example A2-14 / PKE 379: 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-tøt-butyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-tert-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 3; yellow solid (86%). 1 H NMR (DMSO) δ: 1 3.43 (br s, I H), 7.98-7.95 (m, I H), 7.83 (d, J = 2.4 Hz, 2H), 7.75 (d, J = 3.9 Hz, 1 H), 7.49 (d, J = 3.9 Hz, 1 H), 7.40-7.37 (m, 2H), 1 .46 (s, 9H). MS (ESI+) m/z: 400.2 [M + H]+. Mp: 289°C-291 0C.
Example A2-1 5 / PKE 362: 4-teΛf-Butyl-2-(5 '-methyl-[2,2 ' ]bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 5-methylthiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-4- te/t-butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (24%). 1 H NMR (CDCI3) δ: 7.41 (d, J = 3.9 Hz, 1 H), 7.07 (d, J = 3.5 Hz, 1 H), 7.04 (d, J = 3.9 Hz, 1 H), 6.69 (dd, J = 3.5, 1 .0 Hz, 1 H), 4.32 (q, J = 7.1 Hz, 2H), 2.49 (s, 3H), 1 .52 (s, 9H), 1 .37 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 392.2 [M + H]+. Mp: 1 1 7 °C-1 20°C. Example A2-1 6 / PKE 365: 4-te/t-Butyl-2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]-l ,3-thiazole-5- carboxylic acid ethyl ester
Figure imgf000036_0001
Method as for Al -2, using 2-trifluoromethoxy-phenyl boronic acid and 2-(5-bromo-thiophen-2-yl)-4-tert- 5 butyl-thiazole-5-carboxylic acid ethyl ester A2-1 -3; pale yellow needles (90%). 1 H NMR (CDCI3) δ: 7.71 - 7.67 (m, 1 H), 7.54 (d, J = 3.8 Hz, 1 H), 7.40-7.34 (m, 4H), 4.33 (q, J = 7.1 Hz, 2H), 1 .53 (s, 9H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 456.2 [M + H]+. Mp: 1 27 °C-1 300C.
Example A2-1 7 / PKE 356: 4-teΛf-Butyl-2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-te/t-Butyl-2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]- i o thiazole-5-carboxylic acid ethyl ester A2-1 6; yellow solid (75%). 1 H NMR (DMSO) δ: 1 3.44 (br s, I H), 7.92 (m, 1 H), 7.79 (d, J = 4.0 Hz, 1 H), 7.60 (d, J = 4.0 Hz, 1 H), 7.51 (m, 3H), 1 .46 (s, 9H). MS (ESI+) m/z: 428.2 [M + H]+. Mp: 1 78°C-1 800C.
Example A2-1 8 / PKE 370: 2-(5 '-Acetyl-[2,2 ']bithiophenyl-5-yl)-4-teΛf-butyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-acetyl-thiophene-5-boronic acid and 2-(5-bromo-thiophen-2-yl)-4- 15 te/t-butyl-thiazole-S-carboxylic acid ethyl ester A2-1 -3; yellow solid (27%). 1 H NMR (CDCI3) δ: 7.61 (d, J = 4.0 Hz, 1 H), 7.45 (d, J = 4.0 Hz, 1 H), 7.27-7.24 (m, 2H), 4.33 (q, J = 7.1 Hz, 2H), 2.56 (s, 3H), 1 .52 (s, 9H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 420.2 [M + H]+. Mp: 143°C-145°C.
Example A2-1 9 / PKE 377: 4-teΛf-butyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 3-quinoline boronic acid and 2-(5-bromo-thiophene-2-yl)-4-tert-butyl- 20 thiazole-5-carboxylic acid ethyl ester A2-1 -3; yellow solid (27%). 1 H NMR (CDCI3) δ: 9.20 (d, J = 2.3 Hz, I H), 8.31 (d, J = 2.0 Hz, I H), 8.1 0 (d, J = 8.4 Hz, I H), 7.84 (d, J = 8.2 Hz, I H), 7.71 (dt, J = 6.9, 1 .4 Hz, 1 H), 7.58 (m, 1 H), 7.55 (d, J = 3.9 Hz, 1 H), 7.45 (d, J = 3.9 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 1 .55 (s, 9H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 423.2 [M + H]+. Mp: 1 59°C-1 61 °C.
Example A2-20 / PKE 380: 4-teΛf-butyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid: 25 Method as for B2-1 , using 4-teΛf-butyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A2-1 9; yellow solid (92%). λ H NMR (DMSO) δ: 9.39 (s, 1 H), 8.82 (s, 1 H), 8.08 (t, J = 9.3 Hz, 2H), 7.91 (d, J = 3.9 Hz, 1 H), 7.86 (d, J = 3.9 Hz, 1 H), 7.80 (t, J = 7.9 Hz, 1 H), 7.68 (t, J = 7.5 Hz, 1 H), 1 .47 (s, 9H). MS (ESI+) m/z: 395.2 [M + H]+. Mp: 243°C-245°C. Example A2-21 / PKE 345: Sodium 4-te^butyl-2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-thiazole-5- carboxylate (sodium salt of compound PKE 344): Method as for B2-34, using 4-tøt-butyl-2-[5-(3- methoxy-phenyl)-thiophen-2-yl]-thiazole-5-carboxylic acid A2-1 0 / PKE 344; yellow solid (75%). 1 H NMR (DMSO) δ: 7.51 (d, J = 3.9 Hz, 1 H), 7.44 (d, J = 3.9 Hz, 1 H), 7.32 (t, J = 7.8 Hz, 1 H), 7.27 (t, J = 7.8 Hz, 1 H), 7.22 (d, J = 2.0 Hz, 1 H), 6.89 (dd, J = 7.9, 1 .3 Hz, 1 H), 3.81 (s, 3H), 1 .46 (s, 9H). MS (ESI+) m/z: 374.2 [M + H]+. Mp: 31 7 °C-320°C.
Example A2-22 / PKE 31 9: 4-teΛf-Butyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester hydrochloride salt (hydrochloride of compound PKE 31 6): Method as for Bl -60, using 4-te/t-butyl- 2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A2-5 / PKE 31 6; yellow solid (60%). 1 H NMR (DMSO) δ: 9.1 8 (d, J = 1 .8 Hz, I H), 8.68 (dd, J = 5.1 , 1 .1 Hz, I H), 7.51 (dd, J = 8.8, 2.1 Hz, 1 H), 7.91 (d, J = 4.0 Hz, 1 H), 7.86 (d, J = 3.9 Hz, 1 H), 7.77 (dd, J = 8.1 , 5.2 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 1 .46 (s, 9H), 1 .29 (t, J = 7.1 Hz, 3H). Mp: 200°C-203 °C.
4-Trifluoromethyl-l ,3-thiazole series
Example A3-1 -1 : 2-Bromo-4-trifluoromethyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al - 1 -1 , using ethyl 2-amino4-trifluoromethyl-thiazole-5-carboxylate; white solid (71 %). 1 H NMR (CDCI3) δ:
4.39 (q, J = 7.1 Hz, 2H), 1 .37 (t, J = 7.1 Hz, 3H).
Example A3-1 -2 / PKE 470: 2-[2,2 ']Bithiophenyl-5-yl-4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2,2'-bithiophene-5-boronic acid and 2-bromo4-trifluoromethyl- thiazole-5-carboxylic acid ethyl ester A3-1 -1 ; green solid (57%). 1 H NMR (CDCI3) δ: 7.52 (t, J = 2.8 Hz, 1 H), 7.32-7.29 (m, 2H), 7.1 7 (d, J = 2.9 Hz, 1 H), 7.06 (dd, J = 3.9, 2.8 Hz, 1 H), 4.40 (q, J = 5.3 Hz, 2H),
1.40 (t, J = 5.3 Hz, 3H). MS (ESI+) m/z: 390.2 [M + H]+. Mp: 144°C-148°C.
Example A3-2 / PKE 472: 2-[2,2 ']Bithiophenyl-5-yl4-trifluoromethyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[2,2 ']Bithiophenyl-5-yl4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-1 -2; yellow solid (27%). 1 H NMR (DMSO) δ: 7.65 (d, J = 3.0 Hz, 1 H), 7.60 (dd, J = 3.8, 0.7 Hz, 1 H), 7.47 (dd, J = 2.7, 0.6 Hz, 1 H), 7.34 (d, J = 2.9 Hz, 1 H), 7.1 3 (dd, J = 3.7, 2.8 Hz, 1 H). MS (ESI+) m/z: 362.0 [M + H]+. Mp: 250°C-255°C dec.
Example A3-3-1 : 2-Thiophen-2-yl4-trifluoromethyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2-bromo4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-1 -1 ; beige solid (68%). λ H NMR (DMSO) δ: 7.99 (dd, J = 3.8, 1 .0 Hz, 1 H), 7.94 (dd, J = 5.0, 1 .0 Hz, 1 H), 7.26 (dd, J = 5.0, 3.8 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 1 .31 (t, J = 7.0 Hz, 3H). Example A3-3-2: 2-(5-Bromo-thiophen-2-yl)-4-trifluoromethyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -I -2, using 2-thiophen-2-yl-4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-3- 1 ; beige solid (99%). 1 H NMR (DMSO) δ: 7.87 (d, J = 4.1 Hz, I H), 7.42 (d, J = 4.1 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 1 .31 (U = 7.1 Hz, 3H).
Example A3-3-3 / PKE 506: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-4-trifluoromethyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo- thiophen-2-yl)4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester A3-3-2; yellow solid (34%). 1 H NMR (CDCI3) δ: 7.80-7.76 (m, 2 H), 7.57 (d, J = 4.0 Hz, 1 H), 7.53 (s, 1 H), 7.39-7.34 (m, 2H), 7.29 (d, J = 4.0 Hz, I H), 4.41 (q, J = 7.1 Hz, 2H), 1 .41 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 440.0 [M + H]+. Mp: 1 82 °C- 1 84°C.
Example A3-4 / PKE 505: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-4-trifluoromethyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-trifluoromethyl- thiazole-5-carboxylic acid ethyl ester A3-3-3; yellow solid (53%). 1 H NMR (DMSO) δ: 8.00 (m, J = 6.6, 3.9 Hz, 2H), 7.91 (s, 1 H), 7.87 (dd, J = 5.2, 3.7 Hz, 1 H), 7.59 (d, J = 4.0 Hz, 1 H), 7.41 (m, J = 3.7 Hz, 2H). MS (ESI+) m/z: 41 2.1 [M + H]+. Mp: 224°C-226°C.
4-Desmethylated 1 ,3-thiazole series
Example A4-1 / PKE 21 2: 2-[2,2']bithiophenyl-5-yl-thiazole-5-ca rboxylic acid ethyl ester: Method as for Al -1 -2, using 2,2'-bithiophene-5-boronic acid and ethyl 2-bromothiazole-5-carboxylate; green solid (4%). 1 H NMR (CDCI3) δ: 8.31 (s, I H), 7.50 (d, J = 3.9 Hz, I H), 7.29 (m, 2H), 7.1 7 (d, J = 3.9 Hz, I H), 7.05 (dd, J = 5.0, 3.7 Hz, 1 H), 4.38 (q, J = 7.1 Hz, 2H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 322.0 [M + H]+. Mp: 1 1 9°C-1 20°C.
Example A4-2 / PKE 21 1 : 2-[2,2']bithiophenyl-5-yl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[2,2']bithiophenyl-5-yl-thiazole-5-carboxylic acid ethyl ester A4-1 ; orange solid (80%). 1 H NMR (DMSO) δ: 1 3.59 (br s, 1 H), 8.29 (s, 1 H), 7.78 (d, J = 4.0 Hz, 1 H), 7.60 (dd, J = 5.1 , 1 .1 Hz, 1 H), 7.46 (dd, J = 4.0, 1 .1 Hz, 1 H), 7.37 (d, J = 4.0 Hz, 1 H), 7.1 3 (dd, J = 5.1 , 4.0 Hz, 1 H). MS (ESI+) m/z: 294.0 [M + H]+. Mp: >250°C.
Example A4-3-1 : 2-thiophen-2-yl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using ethyl 2-bromo-l ,3-thiazole-5-carboxylate; yellow solid (66%).
1 H NMR (CDCI3) δ: 8.31 (s, I H), 7.61 (dd, J = 3.7, 1 .1 Hz, 1 H), 7.49 (dd, J = 5.1 , 1 .1 Hz, I H), 7.1 2 (dd, J = 5.1 , 3.8 Hz, 1 H), 4.38 (q, J = 7.1 Hz, 2H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 240.0 [M + H]+. Example A4-3-2: 2-(5-bromo-thiophen-2-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -I - 2, using 2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester A4-3-1 ; yellow solid (96%). 1 H NMR (CDCI3) δ: 8.28 (s, 1 H), 7.33 (d, J = 4.0 Hz, 1 H), 7.08 (d, J = 4.0 Hz, 1 H), 4.38 (q, J = 7.1 Hz, 2H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 31 7.9 - 31 9.9 [M + H]+.
Example A4-3-3 / PKE 314: 2-(5-Phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzene boronic acid and 2-(5-bromo-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A4-3-2; yellow solid (25%). 1 H NMR (CDCI3) δ: 8.32 (s, I H), 7.67-7.63 (m, 2H), 7.58 (d, J = 4.0 Hz, 1 H), 7.44-7.39 (m, 2 H), 7.36 (d, J = 7.1 Hz, 1 H), 7.32 (d, J = 4.0 Hz, 1 H), 4.38 (q, J = 7.1 Hz, 2H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 31 6.1 [M + H]+. Mp: 1 33°C-1 35°C.
Example A4-4 / PKE 31 5: 2-(5-Phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester A4-3-3; yellow solid (83%). 1 H NMR (DMSO) δ: 8.29 (s, 1 H), 7.84 (d, J = 4.0 Hz, 1 H), 7.74 (dd, J = 7.0, 1 .4 Hz, 2H), 7.62 (d, J = 4.0 Hz, 1 H), 7.48-7.42 (m, 2H), 7.37 (td, J = 7.1 , 2.5 Hz, 1 H). MS (ESI+) m/z: 288.0 [M + H]+. Mp: 243°C- 245°C.
Example A4-5 / PKE 371 : 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)- thiazole-5-carboxylic acid ethyl ester A4-3-2; yellow solid (1 6%). 1 H NMR (CDCI3) δ: 8.33 (s, I H), 7.82- 7.75 (m, 2H), 7.55 (d, J = 4.0 Hz, 1 H), 7.50 (d, J = 0.6 Hz, 1 H), 7.38-7.33 (m, 2H), 7.28 (d, J = 3.8 Hz, I H), 4.39 (q, J = 7.1 Hz, 2H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 372.1 [M + H]+. Mp: 2000C- 2030C.
Example A4-6 / PKE 214: Sodium 2-[2,2 ' ]bithiophenyl-5-yl-thiazole-5-carboxylate (sodium salt of compound PKE 21 1 ): Method as for B2-34, using 2-[2,2']b ithiophenyl-5-yl-thiazole-5-carboxylic acid A4-2 / PKE 21 1 ; yellow solid (83%). 1 H NMR (DMSO) δ: 7.71 (br s, I H), 7.51 (br s, I H), 7.40 (br s , I H), 7.29 (br s, 2H), 7.1 0 (br s, 1 H). MS (ESI+) m/z: 393.9 [M + H]+. Mp: > 250°C.
Other thiazole substitutions
Example A5-1 -1 : 2-Bromo-l ,3-thiazole4-carboxylic acid ethyl ester: Method as for Al -1 -1 , using 2- amino-thiazole-4-carboxylic acid ethyl ester; white solid (55%). 1 H NMR (CDCI3) δ: 8.1 2 (s, I H), 4.43 (q, J = 7.1 Hz, 2H), 1 .40 (t, J = 7.1 Hz, 3H).
Example A5-1 -2 / PKE 453: 2-[2,2 ']Bithiophenyl-5-yl-thiazole-4-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2,2'-bithiophene-5-boronic acid and 2-bromo-thiazole-4-carboxylic acid ethyl ester; greenish solid (43%). 1 H NMR (CDCI3) δ: 8.07 (s, I H), 7.49 (d, J = 4.0 Hz, I H), 7.28 (td, J = 5.1 , 1 .2 Hz, 2H), 7.15 (d, J = 3.7 Hz, IH), 7.05 (dd, J = 5.1, 3.6 Hz, 1 H), 4.44 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 322.1 [M + H]+. Mp: 89°C-910C.
Example A5-2 / PKE 456: 2-[2,2']Bithiophenyl-5-yl-thiazole4-carboxylic acid: Method as for B2-1, using 2-[2,2']bithiophenyl-5-yl-thiazole-4-carboxylic acid ethyl ester A5-1-2; pale yellow solid (100%).1H NMR (DMSO) δ: 8.42 (s, 1 H), 7.72 (d, J = 4.0 Hz, 1 H), 7.61 (dd, J = 5.0, 1.0 Hz, 1 H), 7.48 (dd, J = 3.6, 1.1 Hz, IH), 7.37 (d, J =4.0 Hz, IH), 7.14 (dd, J = 5.0, 3.7 Hz, IH). MS (ESI+) m/z: 294.0 [M + H]+. Mp: 219°C-221°C.
Example A5-3 / PKE 448: 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole: Method as for Bl-M, using 2- bromo-4-methyl-l,3-thiazole and 2,2'-bithiophenyl-5-boronic acid; beige solid (30%).1H NMR (CDCI3) δ: 8.62 (s, 1 H), 7.24 (dd, J = 5.1 , 1.1 Hz, 1 H), 7.20 (dd, J = 3.6, 1.1 Hz, 1 H), 7.13 (d, J = 3.8 Hz, 1 H), 7.04 (dd, J = 3.8, 1.6 Hz, 1 H), 7.02 (d, J = 3.6 Hz, 1 H), 2.64 (s, 3H). MS (ESI+) m/z: 264.0 [M + H]+. Mp: 83°C-85°C.
Example A5-4-1: 4-Methyl-2-thiophen-2-yl-l,3-thiazole: Method as for Bl-M, using 2-bromo4-methyl- 1,3-thiazole; colourless oil (57%).1H NMR (CDCI3) δ: 8.62 (s, IH), 7.36 (dd, J = 5.1, 1.2 Hz, IH), 7.15 (dd, J = 3.6, 1.2 Hz, 1 H), 7.09 (dd, J = 5.1, 3.6 Hz, 1 H), 2.62 (s, 3H).
Example A5-4-2: 2-(5-Bromo-thiophen-2-yl)-4-methyl-l,3-thiazole: Method as for BM -2, using 4-methyl- 2-thiophen-2-yl-thiazole A5-4-1; beige solid (88%).1H NMR (DMSO) δ: 9.00 (s, 1 H), 7.30 (d, J = 3.9 Hz, IH), 7.13 (d, J = 3.9 Hz, IH).
Example A5-4-3 / PKE 489: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazole: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole A5-4-2; yellow solid (67%).1H NMR (CDCI3) δ: 8.62 (s, 1 H), 7.81-7.72 (m, 2H), 7.43 (s, IH), 7.39-7.29 (m, 2H), 7.26 (d, J = 2.4 Hz, IH), 7.10 (d, J = 3.8 Hz, IH), 2.67 (s, 3H). MS (ESI+) m/z: 314.0 [M + H]+. Mp: 145°C-148°C.
Example A5-5 / PKE 462: 2-[2,2']bithiophenyl-5-yl-5-methyl-thiazole: Method as for Bl-M, using 2- bromo-5-methyl-l ,3-thiazole and 2,2'-bithiophenyl-5-boronic acid; brown solid (36%).1H NMR (CDCI3) δ: 7.40 (s, 1 H), 7.31 (d, J = 3.7 Hz, 1 H), 7.26 (s, 1 H), 7.23 (d, J = 3.9 Hz, 1 H), 7.10 (d, J = 3.7 Hz, 1 H), 7.03 (t, J = 4.6 Hz, 1 H), 2.48 (s, 3H). MS (ESI+) m/z: 264.0 [M + H]+. Mp: 73°C-75°C.
Example A5-6-1: 5-Methyl-2-thiophen-2-yl-l ,3-thiazole: Method as for Bl-M, using 2-bromo-5-methyl- 1 ,3-thiazole; colourless oil (74%).1H NMR (CDCI3) δ: 7.43-7.39 (m, 2H), 7.35 (dd, J = 5.1, 1.1 Hz, IH), 7.05 (dd, J = 5.0, 3.7 Hz, 1 H), 2.48 (s, 3H). MS (ESI+) m/z: 182.0 [M + H]+. Example A5-6-2: 2-(5-Bromo-thiophen-2-yl)-5-methyl-l ,3-thiazole: Method as for Bl -1 -2, using 5-methyl- 2-thiophen-2-yl-thiazole A5-6-1 ; yellow solid (58%). λ H NMR (DMSO) δ: 7.50 (s, 1 H), 7.41 (d, J = 4.0 Hz, 1 H), 7.27 (d, J = 4.0 Hz, 1 H), 2.46 (s, 3H). MS (ESI+) m/z: 260.0-261 .9 [M + H]+.
Example A5-6-3 / PKE 488: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-5-methyl-thiazole: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-5-methyl-thiazole A5-6-2; yellow solid (57%). 1 H NMR (CDCI3) δ: 7.80-7.74 (m, 2 H), 7.44 (d, J = 5.3 Hz, 1 H), 7.43 (m, 1 H), 7.39 (d, J = 4.0 Hz, 1 H), 7.33 (dt, J = 5.4, 1 .5 Hz, 2H), 7.24 (d, J = 4.0 Hz, 1 H), 2.51 (d, J = 1 .3 Hz, 3H). MS (ESI+) m/z: 314.1 [M + H]+. Mp: 1 60°C-1 63°C.
Example A5-7-1 : 2-Bromo-5-methyl-l ,3-thiazole-4-carboxylic acid ethyl ester: Method as for Al -1 -1 , us- ing 2-amino-5-methyl-thiazole4-carboxylic acid ethyl ester; yellow solid (83%). 1 H NMR (CDCI3) δ: 4.33 (q, J = 7.1 Hz, 2H), 2.71 (s, 3H), 1 .36 (t, J = 7.1 Hz, 3H).
Example A5-7-2 / PKE 540: 2-[2,2 ']Bithiophenyl-5-yl-5-methyl-l ,3-thiazole-4-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2-bromo-5-methyl-thiazole-4-carboxylic acid ethyl ester A5-7-1 and 2,2'- bithiophenyl-5-boronic acid; yellow solid (84%). λ H NMR (CDCI3) δ: 7.47 (d, J = 4.0 Hz, 1 H), 7.28 (ddd, J = 6.5, 5.1 , 1 .5 Hz, 2H), 7.1 5 (d, J = 4.0 Hz, I H), 7.05 (dd, J = 5.1 , 3.8 Hz, I H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 336.1 [M + H]+. Mp: 141 0C-144°C.
Example A5-8 / PKE 541 : 2-[2,2 ']Bithiophenyl-5-yl-5-methyl-l ,3-thiazole4-carboxylic acid: Method as for B2-1 , using 2-[2,2 ' ]bithiophenyl-5-yl-5-methyl-thiazole-4-carboxylic acid ethyl ester A5-7-2; yellow solid (78%). λ H NMR (DMSO) δ: 1 3.38 (br s, 1 H), 7.75 (d, J = 4.0 Hz, 1 H), 7.63 (dd, J = 5.0, 1 .0 Hz, I H), 7.49 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.38 (d, J = 4.0 Hz, 1 H), 7.14 (dd, J = 5.1 , 3.8 Hz, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 308.0 [M + H]+. Mp: 242 °C-245°C.
Example A5-9 / PKE 466: 2-[2,2']-Bithophenyl-5-yl-benzothiazole: Method as for Bl -1 -1 , using 2,2'- bithiophenyl-5-boronic acid and 2-bromo-benzothiazole; yellow solid (42%). 1 H NMR (CDCI3) δ: 8.02 (d, J = 8.1 Hz, 1 H), 7.84 (d, J = 7.8 Hz, 1 H), 7.55 (d, J = 3.9 Hz, 1 H), 7.47 (dt, J = 8.1 , 1 .1 Hz, 1 H), 7.36 (dt, J = 8.1 , 1 .0 Hz, 1 H), 7.29 (d, J = 4.3 Hz, 2H), 7.1 9 (d, J = 3.9 Hz, 1 H), 7.06 (t, J = 4.3 Hz, 1 H). MS (ESI+) m/z: 300.1 [M + H]+. Mp: 1 600C-1 68°C.
Example A5-1 0-1 : 2-Bromo-6-methoxy-l ,3-benzo[d]thiazole: Method as for Al -1 -1 , using 2-amino-6- methoxy-benzothiazole; brown solid (26%). λ H NMR (CDCI3) δ: 7.86 (d, J = 6.8 Hz, 1 H), 7.25 (d, J = 1 .9 Hz, 1 H), 7.06 (dd, J = 6.7, 1 .9 Hz, 1 H), 3.87 (s, 3H). MS (ESI+) m/z: 243.9 - 245.9 [M + H]+.
Example A5-1 0-2 / PKE 463: 2-[2,2 ']Bithiophenyl-5-yl-6-methoxy-benzothiazole: Method as for Bl -1 -1 , using 2,2'-bithiophenyl-5-boronic acid and 2-bromo-6-methoxy-benzothiazole A5-1 0-1 ; green solid (25%).1H NMR (CDCI3) δ: 7.92 (brs, IH), 7.51 (br s, 1 H), 7.31-7.28 (m, 3H), 7.18 (brs, IH), 7.09 (br s, 1 H), 7.06 (t, J = 4.0 Hz, 1 H), 3.89 (s, 3H). MS (ESI+) m/z: 330.1 [M + H]+. Mp: 195°C-198°C.
Example A5-11-1: 2-Thiophen-2-yl-l,3-benzo[d]thiazole: Method as for Bl-1-1 using 2-bromo- benzothiazole; white solid (74%).
Example A5-11-2: 2-(5-Bromo-thiophen-2-yl)-l,3-benzo[d]thiazole: Method as for Bl-I -2, using 2- thiophen-2-yl-benzothiazole A5-11-1; white solid (97%).
Example A5-11-3 / PKE 527: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-benzothiazole: Method as for Bl-2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-benzothiazole A5-11-2; yellow solid (32%).1H NMR (CDCI3) δ: 8.04 (d, J = 7.7 Hz, 1 H), 7.86 (d, J = 8.0 Hz, 1 H), 7.79 (m, 2H), 7.60 (s, 1 H), 7.52 (m, 3H), 7.33 (m, 3H). MS (ESI+) m/z: 350.5 [M + H]+. Mp: 209°C-213°C.
Example A5-12 / PKE 528: 2-(5-Naphthalen-2-yl-thiophen-2-yl)-benzothiazole: Method as for Bl-2, using naphthalene-2-boronic acid and 2-(5-bromo-thiophen-2-yl)-benzothiazole A5-11-2; yellow solid (47%).1H NMR (CDCI3) δ: 8.14 (s, 1 H), 8.05 (d, J = 8.1 Hz, 1 H), 7.90-7.77 (m, 4H), 7.67 (d, J = 3.9 Hz, IH), 7.63-7.54 (m, IH), 7.53-7.46 (m, 2H), 7.40-7.28 (m, 3H). MS (ESI+) m/z: 343.7 [M + H]+. Mp: 198°C-200°C.
Example A5-13-1: 2-Bromo4-methyl-thiazole-5-carbonitrile: 2-Bromo4-methyl-thiazole-5-carboxylic acid amide B3-11-1 (120 mg, 0.54 mmol) was dissolved in phosphorus oxychloride (8 ml) and reaction mixture was heated at 800C for 3 h. After hydrolysis of the mixture in water (200 ml), the organics were extracted twice with EtOAc. Organic layers were washed twice with water, dried and concentrated in vacuo to afford a pale yellow solid (91%). Mp: 44°C46°C.
Example A5-13-2/ PKE 544: 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carbonitrile
Figure imgf000042_0001
Method as for Bl-1-1, using 2-bromo-4-methyl-thiazole-5-carbonitrile A5-13-1 and 2,2'-bithiophenyl-5- boronic acid; green solid (49%).1H NMR (CDCI3) δ: 7.48 (d, J = 4.0 Hz, IH), 7.32-7.28 (m, 2H), 7.15 (d, J = 4.0 Hz, 1 H), 7.06 (dd, J = 5.0, 3.7 Hz, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 289.0 [M + H]+. Mp: 118°C-121°C. Example A5-14/ PKE 545: 5-(2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazole-5-yl)-2W-tetrazole
Figure imgf000043_0001
.2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazole-5-carbonitrile A5-1 3-2 (50 mg, 0.1 7 mmol), sodium azide (34 mg, 0.51 mmol) and triethylammonium chloride (68 mg, 0.51 mmol) were stirred in dry toluene (1 0 ml)
5 under argon at 900C for 1 8 h. After concentration in vacuo, the residue was dissolved in EtOAc and extracted with aqueous ammonia (2x1 0 ml). The basic aqueous layer was acidified with concentrated
HCI until pH<2. The precipitate formed was filtered off, washed with water and dried to afford an ochre solid (1 9%). 1 H NMR (acetone) δ: 7.68 (d, J = 4.0 Hz, 1 H), 7.53 (d, J = 5.1 Hz, I H), 7.45 (d, J = 2.6 Hz,
1 H), 7.35 (d, J = 3.9 Hz, 1 H), 7.14 (dd, J = 5.0, 3.7 Hz, 1 H), 2.77 (s, 3H). MS (ESI+) m/z: 332.0 [M + i o H]+.
Terminal thiophene modifications
5-(ethoxycarbonyl)-l ,3-thiazole derivatives (ester series)
Example Bl -1 -1 / PKE 253: 4-methyl-2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester: To a mixture of thiophene-2-boronic acid (0.32 g, 2.5 mmol), 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester
15 (0.56 g, 2.25 mmol), and powdered CsF (0.76 g, 5.0 mmol ) in 1 0 ml of DME degassed under argon was added Pd(PPh3J4 (87 mg, 0.075 mmol). The reaction mixture was degassed under argon and heated up to reflux for 1 8 h. The reaction mixture was then diluted with methylene chloride and water and the organic layer was dried and concentrated. Chromatography of the residue on silica gel (cyclo- hexane-EtOAc: 80/20) afforded a pale grey powder (75%). 1 H NMR (CDCI3) δ: 7.59 (dd, J = 3.7, 1 .1
20 Hz, 1 H), 7.46 (dd, J = 5.0, 1 .1 Hz, 1 H), 7.1 0 (dd, J = 5.0, 3.8 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 254.0 [M + H]+. Mp: 55°C-57 °C dec.
Example Bl -1 -2 / PKE 291 : 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: A mixture of 4-methyl-2-thiophen-2-yl-thiazole-5-carboxylic acid ethyl ester Bl -1 -1 (0.8 g, 3.1 6 mmol) and bromine (0.55 g, 3.48 mmol) in 30 ml of glacial acetic acid was heated at 800C for two hours. The 25 reaction mixture was poured into 200 ml of water and the precipitate formed was filtered off and dried to give a pale green solid (94 %). ] H NMR (CDCI3) δ: 7.30 (d, J = 4.0 Hz, 1 H), 7.05 (d, J = 4.0 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.72 (s, 3H), 1 .37 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 332.0 - 334.0 [M + H]+. Mp: 73°C-75°C. Example Bl -1 -3 / PKE 290: 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid: To a solution of 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (50 mg, 0.1 5 mmol) dissolved in a 8/2 mixture of methanol-water (25 ml) was added lithium hydroxide monohydrate (25 mg, 0.60 mmol). The reaction mixture was heated at reflux for 2 hours. The solvent was evaporated in vacuo, water (25 ml) was added, and the solution extracted with CH2CI2 (3 x 1 5 ml). The aqueous phase was acidified to pH 1 with cone. HCI and extracted twice with EtOAc. The organic phases were collected, dried (Na2SO4), filtered and evaporated in vacuo to afford a white powder (66%). 1 H NMR (DMSO) δ: 7.64 (d, J = 4.0 Hz, 1 H), 7.32 (d, J = 4.0 Hz, 1 H), 2.58 (s, 3H). MS (ESI+) m/z: 304.0-306.0 [M + H]+. Mp: 242 °C-244°C.
Example Bl -2 / PKE 21 2: 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: To a mixture, degassed under argon, of thiophene-2-boronic acid (0.1 1 g, 0.9 mmol), 2-(5-bromo-thiophen-2- yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (0.1 5 g, 0.5 mmol), and powdered CsF (0.28 g, 1 .9 mmol) in 1 0 ml of DME and 3 ml of water was added Pd(PPh3J4 (37 mg, 0.03 mmol). The reaction mixture was degassed under argon and heated up to reflux for 1 8 h. The reaction mixture was then diluted with EtOAc and a saturated aqueous solution of NaHCO3. The organic layer was washed with brine, dried over MgSO4 and concentrated under vacuum. Chromatography of the residue on silica gel (cyclohexane-EtOAc: 80/20) afforded a yellow solid (72%). 1 H NMR (CDCI3) δ: 7.68 (d, J = 4.0 Hz, 1 H), 7.54 (d, J = 4.9 Hz, 1 H), 7.41 (d, J = 2.8 Hz, 1 H), 7.30 (d, J = 4.0 Hz, 1 H), 7.09 (dd, J = 5.0, 3.7 Hz, 1 H), 4.24 (q, J = 7.1 Hz, 2H), 2.60 (s, 3H), 1 .27 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 336.0 [M + H]+. Mp: 1 38°C-141 °C.
Example Bl -3 / PKE 254: 2-[2,3']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using thiophene-3-boronic acid; yellow solid (7%). 1 H NMR (CDCI3) δ: 7.49 (td, J = 7.9, 5.0, 2.9 Hz, 2H), 7.38 (dd, J = 7.9, 2.9 Hz, 1 H), 7.33 (dd, J = 5.0, 1 .2 Hz, 1 H), 7.1 8 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 336.1 [M + H]+. Mp: 86 °C-88°C.
Example Bl -4 / PKE 526: 2-(5-furan-3-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using furane-3-boronic acid; yellow powder (99%). 1 H NMR (CDCI3) δ: 7.75 (s, 1 H), 7.47 (m, 2H), 7.07 (d, J = 3.8 Hz, 1 H), 6.63 (dd, J = 1 .8, 0.8 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 320.1 [M + H]+. Mp: 1 35°C-1 36°C.
Example Bl -5 / PKE 529: 4-methyl-2-[2,2',5',2"]terthiophen-5-yl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using [2,2',5']tert-thiophene-2-boronic acid; yellow solid (69%). 1 H NMR (CDCI3) δ: 7.48 (d, J = 3.9 Hz, 1 H), 7.24 (d, J = 1 .1 Hz, 1 H), 7.20 (dd, J = 3.6, 1 .1 Hz, 1 H), 7.1 7 (d, J = 3.8 Hz, 1 H), 7.1 2 (d, J = 4.0 Hz, 1 H), 7.1 0 (d, J = 3.8 Hz, 1 H), 7.04 (dd, J = 5.1 , 3.6 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 41 8.1 [M + H]+. Mp: 141 0C-144°C. Example Bl -6 / PKE 534: 2-(5-furan-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using furane-2-boronic acid; green solid (73%). 1 H NMR (CDCI3) δ: 7.49 (d, J = 4.0 Hz, 1 H), 7.45 (d, J = 1 .3 Hz, 1 H), 7.21 (d, J = 4.0 Hz, 1 H), 6.61 (d, J = 3.3 Hz, 1 H), 6.47 (dd, J = 3.4, 1 .8 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 320.1 [M + H]+. Mp: 86°C-88°C.
Example Bl -7 / PKE 289: 2-[5-(2-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-methoxyphenyl boronic acid; yellow powder (49%). 1 H NMR (CDCI3) δ: 7.70 (dd, J = 8.3, 1 .7 Hz, 1 H), 7.56 (d, J = 4.0 Hz, 1 H), 7.49 (d, J = 4.0 Hz, 1 H), 7.33 (dt, J = 8.2, 1 .6 Hz, 1 H), 7.04 (dd, J = 8.2, 1 .0 Hz, 2H), 4.34 (q, J = 7.1 Hz, 2H), 3.98 (s, 3H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 360.1 [M + H]+. Mp: 1 1 1 0C-1 1 3°C.
Example Bl -8 / PKE 288: 2-[5-(2-cyano-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-cyanophenyl boronic acid; yellow solid (14%). 1 H NMR (CDCI3) δ: 7.77 (d, J = 7.3 Hz, 1 H), 7.64 (d, J = 5.1 Hz, 2H), 7.59 (d, J = 3.6 Hz, 1 H), 7.47 (t, J = 2.1 Hz, 1 H), 7.25 (m, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 355.1 [M + H]+. Mp: 1 000C-1 020C.
Example Bl -9 / PKE 287: 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methoxyphenyl boronic acid; yellow solid (74%). 1 H NMR (CDCI3) δ: 7.56 (d, J = 8.8 Hz, 2H), 7.51 (d, J = 3.9 Hz, 1 H), 7.1 8 (d, J = 3.9 Hz, 1 H), 7.93 (d, J = 8.8 Hz, 2H), 4.33 (q, J = 7.1 Hz, 2H), 3.84 (s, 3H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 360.1 [M + H]+. Mp: 1 04°C-l 06°C.
Example Bl -1 0 / PKE 301 : 2-[5-(2-fluoro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-fluorophenyl boronic acid; yellow powder (57%). 1 H NMR (CDCI3) δ: 7.66 (dt, J = 8.2, 1 .9 Hz, 1 H), 7.55 (d, J = 4.0 Hz, 1 H), 7.46 (d, J = 4.0 Hz, 1 H), 7.32 (m, 1 H), 7.1 9 (m, 2H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 348.1 [M + H]+. Mp: 1 060C-1 080C.
Example Bl -I l / PKE 297: 4-methyl-2-(5'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 5-methyl-thiophene-2-boronic acid; yellow solid (72%). 1 H NMR (CDCI3) δ: 7.43 (d, J = 3.9 Hz, 1 H), 7.05 (d, J = 3.8 Hz, 1 H), 7.04 (d, J = 1 .0 Hz, 1 H), 6.69 (dd, J = 3.5, 1 .0 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2 H), 2.73 (s, 3H), 2.49 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 350.1 [M + H]+. Mp: 1 01 °C-1 03°C.
Example Bl -1 2 / PKE 302: 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid; yellow solid (43%). 1 H NMR (CDCI3) δ: 7.78 (m, 2H), 7.50 (d, J = 3.9 Hz, 1 H), 7.48 (s, 1 H), 7.36 (m, 2H), 7.26 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 7.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 386.1 [M + H]+. Mp: 1 40°C-142 °C.
Example Bl -1 3 / PKE 303: 2-[5-(3-fluoro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-fluorophenyl boronic acid; yellow solid (67%). 1 H NMR (CDCI3) δ: 7.53 (d, J = 3.9 Hz, 1 H), 7.33 (m, 3H), 7.29 (d, J = 3.9 Hz, 1 H), 7.03 (dt, J = 4.3, 1 .8 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 348.1 [M + H]+. Mp: H l 0C- 1 1 3°C.
Example Bl -14 / PKE 304: 2-(5-benzofuran-2-yl-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzofurane-2-boronic acid; yellow powder (31 %). 1 H NMR (CDCI3) δ: 7.57 (d, J = 8.2 Hz, 1 H), 7.54 (d, J = 4.0 Hz, 1 H), 7.50 (d, J = 8.2 Hz, 1 H), 7.44 (d, J = 4.0 Hz, 1 H), 7.31 (dt, J = 7.3, 1 .3 Hz, 1 H), 7.24 (m, 1 H), 6.97 (s, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 370.1 [M + H]+. Mp: 149°C-1 51 0C.
Example Bl -1 5 / PKE 300: 2-[5-(4-fluoro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-fluorophenyl boronic acid; yellow powder (5%). 1 H NMR (CDCI3) δ: 7.60 (dd, J = 8.8, 5.2 Hz, 2H), 7.52 (d, J = 3.9 Hz, 1 H), 7.22 (d, J = 3.9 Hz, 1 H), 7.1 1 (t, J = 8.6 Hz, 2H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 348.1 [M + H]+. Mp: 96 °C-98°C.
Example Bl -1 6 / PKE 299: 4-methyl-2-(5'-acetyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-acetyl-thiophene-5-boronic acid; yellow solid (44%). 1 H NMR (CDCI3) δ: 7.60 (d, J = 4.0 Hz, 1 H), 7.47 (d, J = 4.0 Hz, 1 H), 7.28 (d, J = 4.0 Hz, 1 H), 7.23 (d, J = 4.0 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 2.53 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 378.1 [M + H]+. Mp: 89°C-91 °C.
Example Bl -1 7 / PKE 31 7: 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 2-trifluoromethoxy-benzene boronic acid; yellow solid (62%). λ H NMR (CDCI3) δ: 7.68 (d, J = 5.6 Hz, 1 H), 7.56 (d, J = 4.0 Hz, 1 H), 7.41 (d, J = 4.0 Hz, I H), 7.36 (m, 3H), 4.37 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 414.1 [M + H]+. Mp: 88°C-90°C.
Example Bl -1 8 / PKE 31 8: 2-[5-(3-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 3-trifluoromethoxy-benzene boronic acid; yellow powder (30%). 1 H NMR (CDCI3) δ: 7.55 (m, 1 H), 7.54 (d, J = 3.9 Hz, 1 H), 7.47 (br s, 1 H), 7.43 (t, J = 8.1 Hz, 1 H), 7.32 (d, J = 3.9 Hz, 1 H), 7.1 9 (m, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 414.1 [M + H]+. Mp: 980C-I OO0C.
Example Bl -1 9 / PKE 322: 2-[5-(4-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-trifluoromethyl benzene boronic acid ; yellow powder (33%). 1 H NMR (CDCI3) δ: 7.74 (d, J = 8.2 Hz, 2H), 7.66 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 3.4 Hz, 1 H),
7.37 (d, J = 3.5 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 398.1 [M + H]+. Mp: 147 °C-149°C.
Example Bl -20 / PKE 324: 2-(5-benzo[ l ,3]dioxol-5-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3,4-methylenedioxy benzene boronic acid; yellow solid (44%). 1 H NMR (CDCI3) δ: 7.52 (d, J = 3.8 Hz, 1 H), 7.1 6 (d, J = 3.8 Hz, 1 H), 7.1 6 (s, 1 H), 7.1 3 (dd, J = 9.7, 1 .8 Hz, 1 H), 6.84 (d, J = 8.0 Hz, 1 H), 6,01 (s, 2H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 374.1 [M + H]+. Mp: 1 29°C-1 300C.
Example Bl -21 / PKE 325: 2-[5-(2,3-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2,3-dimethoxyphenyl boronic acid; yellow solid (34%). 1 H NMR (CDCI3) δ: 7.56 (d, J = 3.8 Hz, 1 H), 7.48 (d, J = 3.9 Hz, 1 H), 7.28 (d, J = 8.0 Hz, 1 H), 7.08 (t, J =
8.1 Hz, 1 H), 6.88 (d, J = 8.0 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 3.90 (s, 3H), 3.89 (s, 3H), 2.76 (s, 3H),
1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 390.2 [M + H]+. Mp: 1 1 9°C-1 21 °C.
Example Bl -22 / PKE 326: 2-[5-(2-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-trifluoromethyl-benzene boronic acid; yellow solid (42%). 1 H NMR (CDCI3) δ: 7.78 (d, J = 7.4 Hz, 1 H), 7.57-7.53 (m, 4H), 7.1 1 (d, J = 5.6 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 398.1 [M + H]+. Mp: 75°C-76°C.
Example Bl -23 / PKE 327: 2-[5-(3-acetyl-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-acetylphenyl boronic acid; yellow powder (1 3%). 1 H NMR (CDCI3) δ: 8.22 (s, 1 H), 7.92 (d, J = 7.7 Hz, 1 H), 7.82 (d, J = 8.2 Hz, 1 H), 7.57 (d, J = 3.9 Hz, 1 H), 7.52 (t, J = 7.8 Hz, 1 H), 7.38 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2 H), 2.76 (s, 3H), 2.65 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 372.2 [M + H]+. Mp: 1 61 0C-1 63°C.
Example Bl -24 / PKE 333: 4-methyl-2-(5-pyrimidin-5-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using pyrimidine-5-boronic acid; yellow powder (25%). 1 H NMR (CDCI3) δ: 9.1 7 (s, 1 H), 8.99 (s, 2H), 7.59 (d, J = 4.0 Hz, 1 H), 7.41 (d, J = 4.0 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 332.1 [M + H]+. Mp: 1 59°C-1 61 0C. Example Bl -25 / PKE 334: 2-[5-(3,4-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3,4-dimethoxyphenyl boronic acid; yellow solid (38%). 1 H NMR (CDCI3) δ: 7.52 (d, J = 3.9 Hz, 1 H), 7.23 (dd, J = 8.4, 2.1 Hz, 1 H), 7.22 (d, J = 3.9 Hz, 1 H), 7.1 2 (d, J = 2.1 Hz, 1 H), 6.90 (d, J = 8.4 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 3.94 (s, 3H), 3.92 (s, 3H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 390.2 [M + H]+. Mp: 1 37 °C-1 38°C.
Example Bl -26 / PKE 349: 2-[5-(3-formyl-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-formylphenyl boronic acid; yellow powder (1 6%). 1 H NMR (CDCI3) δ: 1 0.06 (s, I H), 8.14 (s, I H), 7.87 (d, J = 7.8 Hz, 1 H), 7.83 (d, J = 7.7 Hz, 1 H), 7.59 (d, J = 7.7 Hz, 1 H), 7.57 (d, J = 3.2 Hz, 1 H), 7.39 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 358.2 [M + H]+. Mp: 141 0C-143°C.
Example Bl -27 / PKE 351 : 2-[5-(2,4-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2,4-dimethoxyphenyl boronic acid; yellow solid (64%). 1 H NMR (CDCI3) δ: 7.60 (dd, J = 7.7, 1 .2 Hz, 1 H), 7.53 (d, J = 4.0 Hz, 1 H), 7.38 (d, J = 4.0 Hz, 1 H), 6.57 (d, J = 2.4 Hz, 1 H), 6.54 (s, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 3.95 (s, 3H), 3.85 (s, 3H), 2.75 (s, 3H), 1 .38 (U = 7.1 Hz, 3H). MS (ESI+) m/z: 390.2 [M + H]+. Mp: 1 22 °C-1 24°C.
Example Bl -28 / PKE 363: 2-[5-(3-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-methoxyphenyl boronic acid; green solid (55%). 1 H NMR (CDCI3) δ: 7.54 (d, J = 3.9 Hz, 1 H), 7.28-7.25 (m, 3H), 7.1 6 (m, 1 H), 6.89 (dd, J = 8.1 , 0.8 Hz, 1 H),
4.34 (q, J = 7.1 Hz, 2H), 3.85 (s, 3H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 360.1 [M + H]+. Mp: 1 28°C-1 30°C.
Example Bl -29 / PKE 378: 4-methyl-2-(5-quinolin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using quinoline-3-boronic acid; yellow powder (1 1 %). 1 H NMR (CDCI3) δ: 9.21 (s, 1 H), 8.33 (s, 1 H), 8.1 1 (d, J = 8.3 Hz, 1 H), 7.85 (d, J = 8.0 Hz, 1 H), 7.73 (t, J = 7.0 Hz, 1 H), 7.58 (m, 2H), 7.48 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 381 .2 [M + H]+. Mp: 144°C-146°C.
Example Bl -30 / PKE 373: 2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 4-trifluoromethoxy-benzene boronic acid; yellow powder (32%). 1 H NMR (CDCI3) δ: 7.64 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 4.0 Hz, 1 H), 7.28-7.25 (m, 3H),
4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 414.1 [M + H]+. Mp: 1 27 °C-1 29°C.
Example Bl -31 / PKE 372: 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using naphthalene-2-boronic acid; yellow powder (26%). 1 H NMR (CDCI3) δ: 8.10 (s, 1 H), 7.84-7.81 (m, 3H), 7.74 (dd, J = 8.6, 1.6 Hz, 1 H), 7.57 (d, J = 3.9 Hz, 1 H), 7.49 (m, 2H), 7.42 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.77 (s, 3H), 1.39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 380.2 [M + H]+. Mp: 1200C-I 22°C.
Example Bl -32 / PKE 393: 2-[5-(3-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-trifluoromethyl-benzene boronic acid; yellow powder (67%). 1 H NMR (DMSO) δ: 8.05 (s, 1 H), 8.03 (d, J = 8.6 Hz, 1 H), 7. 86 (d, J = 4.0 Hz, 1 H), 7.71 (d, J = 4.0 Hz, I H), 7.68 (m, 2H), 4.27 (q, J = 7.1 Hz, 2H), 2.63 (s, 3H), 1 .29 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 398.1 [M + H]+. Mp: 1 1 5°C-1 1 7 °C.
Example Bl -33 / PKE 394: 4-methyl-2-(4'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-thiophene-2-boronic acid; yellow powder (21 %). 1 H NMR (CDCI3) δ: 7.45 (d, J = 3.9 Hz, 1 H), 7.1 1 (d, J = 3.9 Hz, 1 H), 7.07 (d, J = 0.9 Hz, 1 H), 6.86 (s, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 2.27 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 350.1 [M + H]+. Mp: 87 °C-89°C.
Example Bl -34 / PKE 388: 4-methyl-2-(5-m-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-methylbenzene boronic acid; yellow powder (46%). 1 H NMR (CDCI3) δ: 7.52 (d, J = 3.9 Hz, 1 H), 7.45 (s, 1 H), 7.42 (m, 1 H), 7.28 (m, 2H), 7.1 6 (m, 1 H), 4.32 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.39 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 344.1 [M + H]+. Mp: 91 0C- 93°C.
Example Bl -35 / PKE 389: 4-methyl-2-(5-naphthalen-l -yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using naphthalene-1 -boronic acid; yellow powder (57%). 1 H NMR (CDCI3) δ: 8.27 (m, 1 H), 7.92 (m, 2H), 7.64 (d, J = 3.8 Hz, 1 H), 7.58 (d, J = 7.0 Hz, 1 H), 7.53 (m, 3H), 7.25 (d, J = 3.8 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.77 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 380.1 [M + H]+. Mp: 1 1 3°C-1 1 5°C.
Example Bl -36 / PKE 390: 4-methyl-2-(5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methylbenzene boronic acid; yellow powder (77%). 1 H NMR (CDCI3) δ: 7.54 (d, J = 3.7 Hz, 1 H), 7.52 (m, 2H), 7.24 (m, 2H), 7.23 (d, J = 7.9 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.38 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 344.1 [M + H]+. Mp: 1 070C-1 09°C.
Example Bl -37 / PKE 391 : 4-methyl-2-(5-o-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-methylbenzene boronic acid; yellow powder (48%). 1 H NMR (CDCI3) δ: 7.56 (d, J = 3.8 Hz, 1 H), 7.43 (d, J = 6.6 Hz, 1 H), 7.29-7.24 (m, 3H), 7.06 (d, J = 3.8 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.47 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 344.1 [M + H]+. Mp: 66 °C-68°C. Example Bl -38 / PKE 392: 4-methyl-2-(3'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-methyl-thiophene-2-boronic acid; yellow powder (33%). 1 H NMR (CDCI3) δ: 7.51 (d, J = 3.9 Hz, 1 H), 7.1 9 (d, J = 5.0 Hz, 1 H), 7.1 2 (d, J = 3.6 Hz, 1 H), 6.90 (d, J = 5.1 Hz, I H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.45 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 350.1 [M + H]+. Mp: 84°C-86°C.
Example Bl -39 / PKE 398: 4-methyl-2-(5'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-phenyl-thiophene-5-boronic acid; yellow powder (54%). 1 H NMR (CDCI3) δ: 7.60 (d, J = 7.3 Hz, 2H), 7.48 (d, J = 3.9 Hz, 1 H), 7.40 (t, J = 7.1 Hz, 2H), 7.32 (d, J = 7.3 Hz, 2H), 7.25 (s, 1 H), 7.1 7 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 41 2.2 [M + H]+. Mp: 1 24°C-1 26°C dec.
Example Bl -40 / PKE 401 : 2-[5-(2-chloro-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-chloro-benzene boronic acid; yellow powder (24%). 1 H NMR (CDCI3) δ: 7.56 (dd, J = 6.2, 4.0 Hz, 2H), 7.50 (dd, J = 5.8, 2.4 Hz, 1 H), 7.38 (d, J = 3.9 Hz, 1 H), 7.31 (m, 2H), 4.36 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .29 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 364.2 - 366.2 [M + H]+. Mp: 73°C-76°C.
Example Bl -41 / PKE 425: 2-[5-(2,4-dimethyl-thiazol-5-yl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2,4-dimethyl-5-(4,4,5,5-tetramethyl-l ,3,2-dioxoborolan-2-yl)- 1 ,3-thiazole; yellow powder (46%). 1 H NMR (CDCI3) δ: 7.49 (d, J = 3.9 Hz, I H), 7.05 (d, J = 3.9 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 2.67 (s, 3H), 2.59 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 365.2 [M + H]+. Mp: 1 28°C-1 300C.
Example B142 / PKE 427: 4-methyl-2-[5-(4-methyl-2-phenyl-thiazol-5-yl)-thiophen-2-yl]-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-5-(4,4,5,5-tetramethyl-l ,3,2-dioxoborolan- 2-yl)-2-phenyl-l ,3-thiazole; yellow powder (86%). λ H NMR (CDCI3) δ: 7.93-7.90 (m, 2H), 7.52 (d, J = 4.0 Hz, 1 H), 7.45-7.42 (m, 3H), 7.14 (d, J = 3.9 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.69 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 427.2 [M + H]+. Mp: 1 600C-1 62 °C.
Example Bl -43 / PKE 428: 4-methyl-2-(4'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-phenyl-thiophene-5-boronic acid; yellow powder (65%). 1 H NMR (CDCI3) δ: 7.60 (d, J = 1 .4 Hz, 1 H), 7.58 (br s, 1 H), 7.53 (d, J = 1 .4 Hz, 1 H), 7.48 (d, J = 4.0 Hz, 1 H), 7.45-7.38 (m, 3H), 7.35 (m, 1 H), 7.1 9 (d, J = 4.0 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 41 2.2 [M + H]+. Mp: 144°C-146°C.
Example Bl -44 / PKE 429: 2-[5-(5-Ethoxycarbonyl4-methyl-thiazol-2-yl)-thiophen-2-yl]-indole-l - carboxylic acid tert-butyl ester: Method as for Bl -2, using /V-Boc-indole-2-boronic acid; yellow oil (35%). 1 H NMR (CDCI3) δ: 8.1 8 (d, J = 8.4 Hz, I H), 7.56 (d, J = 7.8 Hz, 1 H), 7.53 (d, J = 3.9 Hz, 1 H), 7.38 (dt, J = 7.3, 1 .2 Hz, 1 H), 7.26 (dt, J = 7.8, 0.9 Hz, 1 H), 7.1 0 (d, J = 3.9 Hz, 1 H), 6.75 (s, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .49 (s, 9H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 469.3 [M + H]+.
Example Bl -45 / PKE 439: 4-methyl-2-(5-thiazol-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: To a mixture of 2-(tributyl-stannyl)-l ,3-thiazole (0.25 g, 0.7 mmol) and 2-(5-bromo-thiophen-2-yl)-4- methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (0.1 7 g, 0.5 mmol) in 1 0 ml of DME degassed under argon was added Pd(PPh3J4 (22 mg, 0.02 mmol). The reaction mixture was degassed under argon and heated up to reflux for 1 8 h. The reaction mixture was then diluted with methylene chloride and water and the organic layer was dried and concentrated under vacuum. Chromatography of the residue on preparative silica TLC (cyclohexane-EtOAc: 80/20) afforded a yellow powder (42%). 1 H NMR (CDCI3) δ: 7.82 (d, J = 3.1 Hz, 1 H), 7.54 (d, J = 3.9 Hz, 1 H), 7.50 (d, J = 3.9 Hz, 1 H), 7.32 (d, J = 3.1 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 337.1 [M + H]+. Mp: 970C-I OO0C.
Example Bl -46 / PKE 461 : 2-(5-benzo[b]thiophen-5-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-5-boronic acid; yellow solid (41 %). 1 H NMR (CDCI3) δ: 8.08 (d, J = 1 .4 Hz, 1 H), 7.89 (d, J = 8.4 Hz, 1 H), 7.61 (dd, J = 8.4, 1 .7 Hz, 1 H), 7.56 (d, J = 3.9 Hz, 1 H), 7.50 (d, J = 5.4 Hz, 1 H), 7.37 (s, 1 H), 7.34 (d, J = 4.0 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 386.2 [M + H]+. Mp: 1 61 0C-1 63°C
Example Bl Al / PKE 493: 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 5-methoxy-benzo[b]thiophene-2-boronic acid; yellow solid (56%). 1 H NMR (CDCI3) δ: 7.64 (d, J = 8.8 Hz, I H), 7.50 (d, J = 4.0 Hz, I H), 7.41 (s, I H), 7.24 (d, J = 4.0 Hz, 1 H), 7.20 (d, J = 2.5 Hz, 1 H), 6.98 (dd, J = 8.8, 2.5 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 3.88 (s, 3H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 41 6.2 [M + H]+. Mp: 1 28°C- 1 300C.
Example Bl -48 / PKE 494: 2-[5-(5-fluoro-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 5-fluoro-benzo[b]thiophene-2-boronic acid; yellow solid (31 %). 1 H NMR (CDCI3) δ: 7.69 (dd, J = 8.7, 4.7 Hz, I H), 7.48 (d, J = 3.9 Hz, 1 H), 7.42 -7.38 (m, 2H), 7.24 (d, J = 4.0 Hz, 1 H), 7.08 (dt, J = 8.8, 2.5 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (U = 7.1 Hz, 3H). MS (ESI+) m/z: 404.1 [M + H]+. Mp: 1 14°C-1 1 7 °C.
Example Bl -49 / PKE 501 : 2-[5-(6-hydroxy-naphthalen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Bl -2, using 6-hydroxy-naphthalene-2-boronic acid; yellow solid (55%). 1 H NMR (CDCI3) δ: 8.03 (s, I H), 7.77 (d, J = 9.1 Hz, I H), 7.71 (s, I H), 7.70 (s, I H), 7.57 (d, J = 3.9 Hz, 1 H), 7.38 (d, J = 3.9 Hz, 1 H), 7.1 5 (s, 1 H), 7.1 2 (d, J = 2.5 Hz, 1 H), 5.08 (br s, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 396.2 [M + H]+. Mp: 1 1 9°C-1 21 °C.
Example Bl -50 / PKE 438: 2-(5-lsoquinolin4-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using lsoquinoline-4-boronic acid 2,2-dimethylpropanediol-l ,3 cyclic ester; yellow solid (72%). 1 H NMR (CDCI3) δ: 9.27 (s, 1 H), 8.65 (s, 1 H), 8.28 (d, J = 7.7 Hz, 1 H), 8.07 (d, J = 7.3 Hz, 1 H), 7.78 (dt, J = 6.9, 1 .5 Hz, 1 H), 7.68 (dt, J = 6.9, 1 .1 Hz, 1 H), 7.67 (d, J = 3.8 Hz, 1 H), 7.31 (d, J = 3.9 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2 H), 2.77 (s, 3H), 1 .4 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 381 .2 [M + H]+. Mp: 1 24°C-1 26°C.
Example Bl -51 / PKE 257: 4-Methyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using phenyl boronic acid; yellow solid (810Zo)- 1 H NMR (CDCI3) δ: 7.65 (td, J = 7.0, 1 .7 Hz, 2H), 7.54 (d, J = 3.8 Hz, 1 H), 7.41 (tt, J = 7.1 , 1 .5 Hz, 2H), 7.35 (td, J = 7.2, 1 .5 Hz, 1 H), 7.30 (d, J = 3.8 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 330.1 [M + H]+. Mp: 1 1 2 °C-1 14°C.
Example Bl -52 / PKE 258: 4-Methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl es- ter: Method as for Bl -2, using pyridine-3-boronic acid; yellow solid (81 %). 1 H NMR (CDCI3) δ: 8.92 (s, 1 H), 8.57 (d, J = 4.9 Hz, 1 H), 7.91 (dd, J= 8.0, 2.5 Hz, 1 H), 7.57 (d, J = 4.0 Hz, 1 H), 7.39-7.34 (m, 2 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 331 .1 [M + H]+. Mp: 1 25 °C-1 27 °C.
Example Bl -53 / PKE 295: 4-Methyl-2-(5-pyridin-4-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl es- ter: Method as for Bl -2, using pyridine-4-boronic acid; yellow solid (21 %). 1 H NMR (DMSO) δ: 8.62 (d, J = 5.8 Hz, 2H), 7.93 (d, J = 3.9 Hz, 1 H), 7.89 (d, J = 4.0 Hz, 1 H), 7.74 (dd, J = 4.6, 1 .6 Hz, 2H), 4.30 (q, J = 7.0 Hz, 2H), 2.66 (s, 3H), 1 .31 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 331 .1 [M + H]+. Mp: 1 58°C- 1 600C.
Example Bl -54/ PKE 260: 4-Methyl-2-[5-(l H-pyrazol4-yl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 1 W-pyrazole-4-boronic acid; yellow solid (14%). 1 H NMR (DMSO) δ: 1 3.06 (br s, 1 H), 7.83 (br s, 1 H), 7.79 (d, J = 4.0 Hz, 1 H), 7.47 (d, J = 4.0 Hz, 1 H), 6.75 (d, J = 2.3 Hz, 1 H), 4.29 (q, J = 7.1 Hz, 2H), 2.65 (s, 3H), 1 .30 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 320.1 [M + H]+. Mp: 206 °C-208°C.
Example Bl -55 / PKE 330: 4-Methyl-2-[5-( 1 -methyl-1 H-pyrazol4-yl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using l -Methyl4-(4,4,5,5-tetramethyl-l ,3,2-dioxaborolan-2-yl)-l H- pyrazole; yellow solid (23%). λ H NMR (DMSO) δ: 8.1 9 (s, 1 H), 7.86 (d, J = 0.8 Hz, 1 H), 7.77 (d, J = 3.8 Hz, 1 H), 7.28 (d, J = 4.0 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 3.87 (s, 3H), 2.63 (s, 3H), 1.29 (t, J = 7.0 Hz, 3H). MS (ESI+) m/z: 334.1 [M + H]+. Mp: 1500C-152°C.
Example Bl -56 / PKE 402: 2-[5-(3-Chloro-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 3-chlorobenzene boronic acid; yellow solid (26%). 1 H NMR (CDCI3) δ: 7.62 (d, J = 1 .9 Hz, 1 H), 7.53 (d, J = 4.0 Hz, 1 H), 7.52-7.49 (m, 1 H), 7.36-7.28 (m, 3H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 364.2 [M + H]+. Mp: 1 18°C- 1 21 0C.
Example Bl -57 / PKE 403: 2-(5-Benzo[b]thiophen-3-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using l -benzothiophen-3-ylboronic acid; yellow solid (28%). 1 H NMR (CDCI3) δ: 8.1 8 (d, J = 7.4 Hz, 1 H), 7.91 (d, J = 7.6 Hz, 1 H), 7.62 (d, J = 3.6 Hz, 2H), 7.45 (qi, J = 7.5 Hz, 2H), 7.34 (d, J = 3.7 Hz, I H), 4.36 (q, J = 7.1 Hz, 2H), 2.77 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 386.2 [M + H]+. Mp: 1 000C-1 040C.
Example Bl -58 / PKE 405: 4-Methyl-2-[5-(5-methyl-benzo[b]thiophen-2-yl)-thiophen-2-yl]-thiazole-5- ca rboxylic acid ethyl ester: Method as for Bl -2, using 5-methylbenzo[b]thiophen-2-boronic acid; yellow solid (31 %). 1 H NMR (CDCI3) δ: 7.67 (d, J = 8.2 Hz, I H), 7.55 (s, 1 H), 7.49 (d, J = 4.0 Hz, I H), 7.41 (s, 1 H), 7.24 (d, J = 4.0 Hz, 1 H), 7.1 6 (d, J = 8.2 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 2.46 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 400.2 [M + H]+. Mp: 144°C-147 °C.
Example Bl -59 / PKE 406: 2-[5-(4-Cyano-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-cyanobenzene boronic acid; yellow solid (30%). 1 H NMR (CDCI3) δ: 7.71 (d, J = 8.5 Hz, 2H), 7.67 (d, J = 8.5 Hz, 2H), 7.54 (d, J = 4.0 Hz, 1 H), 7.39 (d, J = 4.0 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 1 .37 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 355.2 [M + H]+. Mp: 1 76°C-1 80°C.
Example Bl -60 / PKE 261 : 4-Methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester hydrochloride salt (hydrochloride of compound PKE 258): A 4M solution of hydrogen chloride in diox- ane (0.020 ml, 0.080 mmol) was added to a solution of 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole- 5-carboxylic acid ethyl ester Bl -52 / PKE 258 (25 mg, 0.076 mmol) in ethanol (2 ml) and dioxane (1 ml). After 2 hours of stirring at room temperature, the precipitate was filtered off and dried under vacuum to afford a yellow solid (68%). 1 H NMR (DMSO) δ: 9.1 2 (d, J = 5.1 Hz, 1 H), 8.65 (dd, J = 5.1 , 1 .3 Hz, 1 H), 8.36 (dt, J = 7.9, 1 .3 Hz, 1 H), 7.94 (d, J = 4.0 Hz, 1 H), 7.83 (d, J = 4.0 Hz, 1 H), 7.68 (dd, J = 8.1 , 5.1 Hz, 1 H), 4.30 (q, J = 7.1 Hz, 2H), 2.66 (s, 3H), 1 .31 (t, J = 7.1 Hz, 3H). Mp: 250°C-253°C.
Example Bl -61 / PKE 294: 4-Methyl-2-(5-pyridin-4-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester hydrochloride salt (hydrochloride of compound PKE 295): Method as for Bl -60, using 4-methyl-2-(5- pyridin-4-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -53 / PKE 295; yellow solid (68%). 1 H NMR (DMSO) δ: 8.80 (m, 2H), 8.1 5 (m, 2H), 8.14 (d, J = 4.0 Hz, I H), 8.03 (d, J = 4.0 Hz, I H), 4.31 (q, J = 7.1 Hz, 2H), 2.68 (s, 3H), 1 .31 (t, J = 7.1 Hz, 3H). Mp: 240°C-242 °C.
5-carboxy-l ,3-thiazole series
Example B2-1 / PKE 193: 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid: To a solution of 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -2 (1 20 mg, 0.36 mmol) dissolved in a 8/2 mixture of methanol-water (50 ml) was added lithium hydroxide monohydrate (50 mg, 1 .1 9 mmol). The reaction mixture was heated at reflux for 2 hours. The solvent was evaporated in vacuo, water (25 ml) was added, and the solution was washed with CH2Cb (3 x 1 5 ml). The aqueous phase was acidified to pH 1 with concentrated HCI and extracted twice with EtOAc. The organic phases were collected, dried (Na2SO4), filtered and evaporated in vacuo to afford a yellow powder (73%). 1 H NMR (DMSO) δ: 1 3.4 (br s, 1 H), 7.73 (d, J = 3.9 Hz, 1 H), 7.60 (d, J = 4.9 Hz, 1 H), 7.47 (d, J = 2.73 Hz, 1 H), 7.35 (d, J = 3.9 Hz, 1 H), 7.1 2 (t, J = 4.8 Hz, 1 H), 2.60 (s, 3H). MS (ESI+) m/z: 308.1 [M + H]+. Mp: >300°C.
Example B2-2 / PKE 262: 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -52 / PKE 258; yellow needles (98%). λ H NMR (DMSO) δ: 9.1 8 (s, 1 H), 8.69 (d, J = 5.0 Hz, 1 H), 8.51 (d, J = 8.0 Hz, 1 H), 7.89-7.86 (m, 2H), 7.79 (dd, J = 8.0, 5.4 Hz, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 303.1 [M + H]+. Mp: 247 °C-249°C.
Example B2-3 / PKE 292: 4-methyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -51 / PKE 257; yellow solid (44%). MS (ESI+) m/z: 302.1 [M + H]+. Mp: 236°C-238°C
Example B2-4 / PKE 298: 4-methyl-2-(5'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester Bl -1 1 ; yellow needles (65%). λ H NMR (DMSO) δ: 1 3.29 (br s, 1 H), 7.68 (d, J = 3.7 Hz, 1 H), 7.25 (br s, 2H), 6.82 (br s, 1 H), 2.59 (s, 3H), 2.45 (s, 3H). MS (ESI+) m/z: 322.1 [M + H]+. Mp: 238°C-240°C.
Example B2-5 / PKE 31 3: 2-(5'-acetyl-[2,2']bithiophen-5-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5'-acetyl-[2,2']bithiophen-5-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 6; ochre solid (95%). 1 H NMR (DMSO) δ: 7.90 (d, J = 4.0 Hz, 1 H), 7.77 (d, J = 4.0 Hz, 1 H), 7.57 (d, J = 4.0 Hz, 1 H), 7.55 (d, J = 4.0 Hz, 1 H), 3.31 (s, 3H), 2.52 (s, 3H). MS (ESI+) m/z: 350.1 [M + H]+. Mp: 245 °C-247 °C. Example B2-6: / PKE 361 : 2-[5-benzo[b]thiophen-2-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 2; orange solid (95%). 1 H NMR (DMSO) δ: 1 3.36 (br s, I H), 7.98-7.95 (m, I H), 7.86- 7.84 (m, 1 H), 7.82 (s, 1 H), 7.80 (d, J = 4.0 Hz, 1 H), 7.51 (d, J = 4.0 Hz, 1 H), 7.42-7.35 (m, 2H), 2.62 (s, 3H). MS (ESI+) m/z: 358.1 [M + H]+. Mp: 252 °C-255°C.
Example B2-7 / PKE 364: 2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(3-methoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -28; yellow solid (72%). 1 H NMR (DMSO) δ: 1 3.37 (br s, I H), 7.79 (d, J = 3.9 Hz, I H), 7.62 (d, J = 3.9 Hz, 1 H), 7.38-7.29 (m, 2H), 7.26 (s, 1 H), 6.94 (d, J = 7.7 Hz, 1 H), 3.81 (s, 3H), 2.61 (s, 3H). MS (ESI+) m/z: 332.1 [M + H]+. Mp: 269°C-271 0C.
Example B2-8 / PKE 366: 2-[5-(2,4-dimethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(2,4-dimethoxy-phenyl)-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -27; yellow solid (97%). 1 H NMR (DMSO) δ: 1 3.28 (br s, I H), 7.75 (d, J = 8.7 Hz, I H), 7.70 (d, J = 4.1 Hz, 1 H), 7.54 (d, J = 4.1 Hz, 1 H), 6.71 (d, J = 2.4 Hz, 1 H), 6.63 (dd, J = 8.7, 2.4 Hz, I H), 3.94 (s, 3H), 3.81 (s, 3H), 2.61 (s, 3H). MS (ESI+) m/z: 362.1 [M + H]+. Mp: 31 7 °C-31 9°C.
Example B2-9 / PKE 368: 2-(5-benzo[ l ,3]dioxol-5-yl-thiophen-2-yl)-4-methyl-thiazole-5-ca rboxylic acid: Method as for B2-1 , using 2-(5-benzo[ l ,3]dioxol-5-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -20; yellow solid (68%). λ H NMR (DMSO) δ: 7.73 (d, J = 3.9 Hz, 1 H), 7.48 (d, J = 3.9 Hz, 1 H), 7.35 (d, J = 1 .6 Hz, 1 H), 7.22 (dd, J = 8.1 , 1 .6 Hz, 1 H), 6.97 (d, J = 8.1 Hz, 1 H), 6.07 (s, 2H), 2.60 (s, 3H). MS (ESI+) m/z: 346.1 [M + H]+. Mp: 255°C-257 °C.
Example B2-1 0 / PKE 375: 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(4-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -9; yellow solid (93%). 1 H NMR (DMSO) δ: 7.73 (d, J = 3.9 Hz, I H), 7.66 (d, J = 8.8 Hz, 2H), 7.46 (d, J = 3.9 Hz, 1 H), 6.99 (d, J = 8.8 Hz, 2H), 3.79 (s, 3H), 2.60 (s, 3H). MS (ESI+) m/z: 332.1 [M + H]+. Mp: 256°C-258°C.
Example B2-1 1 / PKE 376: 2-[5-(2-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(2-methoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -7; yellow solid (89%). λ H NMR (DMSO) δ: 7.83 (dd, J = 7.8, 1 .4 Hz, 1 H), 7.68 (d, J = 4.0 Hz, 1 H), 7.66 (d, J = 4.0 Hz, 1 H), 7.37 (dt, J = 7.1 , 1 .5 Hz, 1 H), 7.1 7 (d, J = 8.1 Hz, 1 H), 7.04 (t, J = 7.0 Hz, 1 H), 3.95 (s, 3H), 2.62 (s, 3H). MS (ESI+) m/z: 332.0 [M + H]+. Mp: 244°C-246°C.
Example B2-1 2 / PKE 374: 2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-[5-(4-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl -1 8; orange solid (86%). 1 H NMR (DMSO) δ: 7.86 (d, J = 8.8 Hz, 2H), 7.79 (d, J = .4.0 Hz, 1 H), 7.63 (d, J = 4.0 Hz, 1 H), 7.42 (d, J = 8.8 Hz, 2H), 2.61 (s, 3H). MS (ESI+) m/z: 386.1 [M + H]+. Mp: 255°C-258°C.
Example B2-1 3 / PKE 347: 2-[5-(3-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(3-trifluoromethyl-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid ethyl ester Bl -32; yellow solid (94%). 1 H NMR (DMSO) δ: 8.04 (s, I H), 8.03 (m, I H), 7.81 (d, J = 4.0 Hz, 1 H), 7.77 (d, J = 4.0 Hz, 1 H), 7.72-7.66 (m, 2H), 2.61 (s, 3H). MS (ESI+) m/z: 370.1 [M + H]+. Mp: 225°C-227 °C.
Example B2-14 / PKE 382: 4-methyl-2-(5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester BI -36; yellow solid (64%). λ H NMR (DMSO) δ: 1 3.35 (br s, 1 H), 7.75 (d, J = 3.7 Hz, 1 H), 7.61 (d, J = 7.8 Hz, 2H), 7.53 (d, J = 3.7 Hz, I H), 7.24 (d, J = 7.7 Hz, 2H), 2.61 (s, 3H), 2.31 (s, 3H). MS (ESI+) m/z: 31 6.1 [M + H]+. Mp: 1 93 °C-1 95°C.
Example B2-1 5 / PKE 381 : 4-methyl-2-(3'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(3'-methyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester Bl -38; orange solid (93%). 1 H NMR (DMSO) δ: 7.75 (d, J = 4.0 Hz, I H), 7.50 (d, J = 5.1 Hz, I H), 7.24 (d, J = 4.0 Hz, 1 H), 7.01 (d, J = 5.1 Hz, 1 H), 2.60 (s, 3H), 2.38 (s, 3H). MS (ESI+) m/z: 322.2 [M + H]+. Mp: 201 °C-204°C.
Example B2-1 6 / PKE 396: 4-methyl-2-(5'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5'-phenyl-[2,2']bithiophen-5-yl]-thiazole-5-carboxylic acid ethyl ester Bl -39; ochre solid (50%). 1 H NMR (DMSO) δ: 1 3.37 (br s, 1 H), 7.75 (d, J = 3.9 Hz, I H), 7.67 (d, J = 7.5 Hz, 2H), 7.54 (d, J = 3.8 Hz, 1 H), 7.49 (d, J = 3.7 Hz, 1 H), 745-740 (m, 3H), 7.32 (t, J = 7.2 Hz, 1 H), 2.61 (s, 3H). MS (ESI+) m/z: 384.2 [M + H]+. Mp: 248°C-250°C dec.
Example B2-1 7 / PKE 397: 4-methyl-2-(5-naphthalen-l -yl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-naphthalen-l -yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -35; pale yellow solid (50%). 1 H NMR (DMSO) δ: 8.1 9-8.1 7 (m, I H), 8.04 (d, J = 3.8 Hz, I H), 8.00 (s, 1 H), 7.87 (d, J = 3.8 Hz, 1 H), 7.66 (d, J = 6.1 Hz, 1 H), 7.61 -7.55 (m, 3H), 7.40 (d, J = 3.8 Hz, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 352.1 [M + H]+. Mp: 224°C-226°C.
Example B2-1 8 / PKE 404: 2-(5-benzo[b]thiophen-3-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-3-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -57/ PKE 403; yellow solid (65%). 1 H NMR (DMSO) δ: 8.1 9 (d, J = 7.3 Hz, I H), 8.1 7 (s, 1 H), 8.1 0 (d, J = 7.3 Hz, 1 H), 7.87 (d, J = 3.9 Hz, 1 H), 7.59 (d, J = 3.9 Hz, 1 H), 7.52-7.48 (m, 2 H), 2.63 (s, 3H). MS (ESI+) m/z: 358.1 [M + H]+. Mp: 220°C-226°C.
Example B2-1 9 / PKE 407: 4-methyl-2-[5-(5methyl-benzo[b]thiophen-2-yl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-methyl-2-[5-(5methyl-benzo[b]thiophen-2-yl)-thiophen-2-yl]- thiazole-5-carboxylic acid ethyl ester Bl -58 / PKE 405; yellow solid (74%). 1 H NMR (DMSO) δ: 7.81 (d, J = 8.2 Hz, 1 H), 7.67 (s, 1 H), 7.59 (m, 2H), 7.42 (d, J = 3.6 Hz, 1 H), 7.1 7 (d, J = 8.0 Hz, 1 H), 2.55 (s, 3H), 2.38 (s, 3H). MS (ESI+) m/z: 372.2 [M + H]+. Mp: 275°C-285°C.
Example B2-20 / PKE 431 : 4-methyl-2-(4'-phenyl-[2,2']bithiophen-5-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4'-phenyl-[2,2']bithiophen-5-yl)-thiazole-5-carboxylic acid ethyl ester Bl 43; yellow solid (80%). 1 H NMR (DMSO) δ: 1 3.40 (br s, I H), 7.94 (s, I H), 7.92 (s, I H), 7.77- 7.75 (m, 3H), 7.44-7.40 (m, 3H), 7.31 (t, J = 7.1 Hz, 1 H), 2.61 (s, 1 H). MS (ESI+) m/z: 384.1 [M + H]+. Mp: 261 °C-263°C.
Example B2-21 / PKE 430: 4-methyl-2-[5-(4-methyl-2-phenyl-thiazol-5-yl)-thiophen-2-yl]-thiazole-5- carboxylic acid: Method as for B2-1 , using 4-methyl-2-[5-(4-methyl-2-phenyl-thiazol-5-yl)-thiophen-2-yl]- thiazole-5-carboxylic acid ethyl ester Bl 42; yellow solid (75%). 1 H NMR (DMSO) δ: 1 3.42 (br s, I H), 7.92 (m, 2H), 7.82 (d, J = 4.0 Hz, 1 H), 7.50 (m, 3H), 7.38 (d, J = 4.0 Hz, 1 H), 2.62 (s, 3H), 2.61 (s, 3H). MS (ESI+) m/z: 399.2 [M + H]+. Mp: >300°C.
Example B2-22 / PKE 440: 2-[5-(l W-lndol-2-yl)-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-(5-Ethoxycarbonyl4-methyl-thiazol-2-yl)-thiophen-2-yl]-indole-l -carboxylic acid te/t-butyl ester Bl 44; brown solid (47%). 1 H NMR (DMSO) δ: 1 1 .74 (br s, I H), 7.79 (d, J = 4.0 Hz, 1 H), 7.56 (d, J = 4.0 Hz, 1 H), 7.52 (d, J = 7.9 Hz, 1 H), 7.38 (d, J = 8.2 Hz, 1 H), 7.1 3 (dt, J = 8.0, 1 .0 Hz, 1 H), 7.00 (dt, J = 7.8, 0.7 Hz, 1 H), 6.86 (d, J = 1 .4 Hz, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 341 .1 [M + H]+. Mp: 21 7 °C-21 9°C.
Example B2-23 / PKE 449: 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Bl -31 ; pale yellow solid (81 %). 1 H NMR (DMSO) δ: 8.31 (s, 1 H), 7.98 (d, J = 8.4 Hz, 2H), 7.91 (dt, J = 7.8, 1 .5 Hz, 2H), 7.81 (d, J = 3.9 Hz, 1 H), 7.74 (d, J = 3.9 Hz, 1 H), 7.55-7.51 (m, 2H), 2.63 (s, 3H). MS (ESI+) m/z: 352.2 [M + H]+. Mp: 235°C-238°C
Example B2-24 / PKE 451 : 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-[5-(2-trifluoromethoxy-phenyl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl -1 7; yellow solid (1 00%). 1 H NMR (DMSO) δ: 7.95 (d, J = 5.3 Hz, 1 H), 7.85 (d, J = 4.1 Hz, 1 H), 7.64 (d, J = 4.0 Hz, 1 H), 7.53-7.49 (m, 3H), 2.64 (s, 3H). MS (ESI+) m/z: 386.1 [M + H]+. Mp: 1 85°C-1 88°C.
Example B2-25 / PKE 537: 2-(5-furan-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-furan-2-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -6; ochre 5 solid (82%). λ H NMR (DMSO) δ: 1 3.40 (br s, 1 H), 7.78 (m, 2H), 7.41 (br s, 1 H), 6.97 (br s, 1 H), 6.63 (br s, 1 H), 2.59 (s, 3H). MS (ESI+) m/z: 292.0 [M + H]+. Mp: 235°C-237 °C.
Example B2-26/ PKE 452: 2-(5-isoquinolin-4-yl-thiophen-2-yl)4-methyl-l ,3-thiazole-5-carboxylic acid
Figure imgf000058_0001
Method as for B2-1 , using 2-(5-isoquinolin4-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl i o ester Bl -50 / PKE 438; yellow solid (1 00%). 1 H NMR (DMSO) δ: 9.63 (s, I H), 8.75 (s, I H), 8.41 (d, J = 8.1 Hz, 1 H), 8.35 (d, J = 8.5 Hz, 1 H), 8.05 (dt, J = 6.8, 1 .1 Hz, 1 H), 7.99 (d, J = 4.0 Hz, 1 H), 7.94 (t, J = 7.1 Hz, 1 H), 7.61 (d, J = 4.0 Hz, 1 H), 2.65 (s, 3H). MS (ESI+) m/z: 353.2 [M + H]+. Mp: 275°C-279°C.
Example B2-27 / PKE 468: 2-(5-benzo[b]thiophen-5-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid: Method as for Bl -2, using 2-(5-benzo[b]thiophen-5-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid 15 ethyl ester Bl 46; brown solid (84%). λ H NMR (DMSO) δ: 8.26 (d, J = 1 .3 Hz, 1 H), 8.06 (d, J = 8.5 Hz, 1 H), 7.82 (d, J = 5.4 Hz, 1 H), 7.78 (d, J = 3.9 Hz, 1 H), 7.72 (dd, J = 8.4, 1 .4 Hz, 1 H), 7.63 (d, J = 3.9 Hz, 1 H), 7.49 (d, J = 5.4 Hz, 1 H), 2.61 (s, 3H). MS (ESI+) m/z: 358.1 [M + H]+. Mp: 244°C-246°C.
Example B2-28 / PKE 491 : 2-[5-(5-hydroxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: To a solution of 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-
20 5-carboxylic acid ethyl ester Bl -47 (0.04 g, 0.1 0 mmol) dissolved in 5 ml of CH2CI2, was added drop- wise with stirring a I M solution of boron tribromide in CH2Cb (0.25 ml, 0.25 mmol). The reaction mixture was heated at 8O0C for 8 hours and stirred at room temperature for 36 hours. The solvent was evaporated in vacuo, EtOAc was added, and the organic phase was washed with water. The organic phase was then extracted with 1 0% ammonia solution and the basic phase was washed twice with
25 EtOAc. The aqueous phase was acidified to pH 1 with concentrated HCI and extracted twice with EtOAc. The organic phases were collected, dried (Na2SO4), filtered and evaporated in vacuo to afford a brown powder (42%). λ H NMR (DMSO) δ: 9.62 (br s, 1 H), 7.71 (d, J = 8.7 Hz, 1 H), 7.62 (d, J = 3.7 Hz, 1 H), 7.61 (s, 1 H), 7.40 (d, J = 3.9 Hz, 1 H), 7.1 6 (d, J = 2.3 Hz, 1 H), 6.87 (dd, J = 8.7, 2.4 Hz, 1 H), 2.58 (s, 3H). MS (ESI+) m/z: 374.0 [M + H]+. Mp: 239°C-241 0C dec. Example B2-29 / PKE 492: 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for Bl -2, using 2-[5-(5-methoxy-benzo[b]thiophen-2-yl)-thiophen-2-yl]4- methyl-thiazole-5-carboxylic acid ethyl ester Bl 47; yellow solid (71 %). 1 H NMR (DMSO) δ: 7.82 (d, J = 8.8 Hz, 1 H), 7.76 (d, J = 3.9 Hz, 1 H), 7.69 (s, 1 H), 7.46 (d, J = 3.9 Hz, 1 H), 7.35 (d, J = 2.3 Hz, 1 H), 7.00 (dd, J = 8.8, 2.4 Hz, 1 H), 3.80 (s, 3H), 2.61 (s, 3H). MS (ESI+) m/z: 388.1 [M + H]+. Mp: 255°C- 257 °C.
Example B2-30 / PKE 495: 2-[5-(5-fluoro-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for Bl -2, using 2-[5-(5-fluoro-benzo[b]thiophen-2-yl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl 48; brown solid (72%). 1 H NMR (DMSO) δ: 8.00 (dd, J = 8.7, 6.7 Hz, 1 H), 7.78 (d, J = 4.0 Hz, 1 H), 7.77 (s, 1 H), 7.64 (dd, J = 9.6, 2.1 Hz, 1 H), 7.52 (d, J = 3.9 Hz, I H), 7.28-7.22 (dt, J = 9.0, 2.3 Hz, I H), 2.62 (s, 3H). MS (ESI+) m/z: 376.1 [M + H]+. Mp: 239°C- 241 0C.
Example B2-31 / PKE 507: 2-[5-(6-hydroxy-naphthalen-2-yl)-thiophen-2-yl]4-methyl-thiazole-5- carboxylic acid: Method as for Bl -2, using 2-[5-(6-hydroxy-naphthalen-2-yl)-thiophen-2-yl]4-methyl- thiazole-5-carboxylic acid ethyl ester Bl 49; yellow solid (72%). 1 H NMR (DMSO) δ: 1 3.35 (br s, I H), 9.90 (s, 1 H), 8.1 7 (s, 1 H), 7.84-7.71 (m, 4H), 7.63 (d, J = 3.9 Hz, 1 H), 1 .1 3 (s, 1 H), 7.1 3-7.09 (m, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 368.1 [M + H]+. Mp: 264°C-268°C dec.
Example B2-32 / PKE 524: 2-(5-furan-3-ylthiophen-2-yl)4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[5-furan-3-yl-thiophen-2-yl]4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -4; yellow solid (88%). 1 H NMR (DMSO) δ: 8.23 (br s, 1 H), 7.77 (br s, 1 H), 7.73 (br s, 1 H), 7.36 (br s, 1 H), 6.92 (br s, 1 H), 2.60 (s, 3H). MS (ESI+) m/z: 292.0 [M + H]+. Mp: 214°C-21 7 °C.
Example B2-33 / 525: 4-Methyl-2-[2,2',5',2"]tertiophen-5-yl-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-[2,2',5',2"]terthiophen-5-yl-thiazole-5-carboxylic acid ethyl ester Bl -5; orange solid (64%). 1 H NMR (DMSO) δ: 7.73 (d, J = 4.0 Hz, 1 H), 7.55 (dd, J = 5.1 , 1 .0 Hz, 1 H), 7.43 (d, J = 3.8 Hz, 1 H), 7.39-7.35 (m, 2H), 7.30 (d, J = 3.8 Hz, 1 H), 7.1 0 (dd, J = 5.1 , 3.6 Hz, 1 H), 2.60 (s, 3H). MS (ESI+) m/z: 390.0 [M + H]+. Mp: 261 °C-264°C.
Example B2-34 / PKE 209: Sodium 2-[2,2 ' ]bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylate (sodium salt of compound PKE 1 93): To a suspension of 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid B2-1 / PKE 1 93 (430 mg, 1 .4 mmol) in water (20 ml) at room temperature is added sodium hy- droxide (56 mg, 1 .4 mmol). Reaction mixture is stirred at room temperature for 2 h. Solvents are evaporated to dryness to give a yellow solid (69%). 1 H NMR (DMSO) δ: 7.55 (dd, J = 5.1 , 1 .1 Hz, I H), 7.45 (d, J = 3.9 Hz, 1 H), 7.39 (dd, J = 3.6, 1 .1 Hz, 1 H), 7.27 (d, J = 3.9 Hz, 1 H), 7.1 0 (dd, J = 5.1 , 3.7 Hz, 1 H), 2.53 (s, 3H). MS (ESI+) m/z: 308.0 [M + H]+. Mp: >250°C. Example B2-35 / PKE 207: Ammonium 2-[2,2 ']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylate (ammonium salt of compound PKE 1 93): To a suspension of 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5- carboxylic acid B2-1 / PKE 193 in water (50ml) and ethanol (50 ml) at room temperature was added a 30% aqueous solution of ammonia, until dissolution of the precipitate. Solvents were evaporated to dryness to give a yellow solid (63%). 1 H NMR (DMSO) δ: 7.57 (m, 2H), 7.42 (dd, J = 5.5, 0.9 Hz, I H), 7.30 (d, J = 4.0 Hz, 1 H), 7.1 1 (dd, J = 5.0, 3.7 Hz, 1 H), 2.56 (s, 3H). MS (ESI+) m/z: 308.0 [M + H]+. Mp: 225°C-230°C dec.
Example B2-36 / PKE 359: Sodium 4-methyl-2-(5 '-methyl-[2,2 ' ]bithiophenyl-5-yl)-thiazole-5-carboxylate (sodium salt of compound PKE 298): Method as for B2-34, using 4-methyl-2-(5'-methyl-[2,2']bithiophen- 5-yl]-thiazole-5-carboxylic acid B2^ / PKE 298; yellow solid (1 00%). 1 H NMR (DMSO) δ: 7.42 (d, J = 3.9 Hz, 1 H), 7.1 7 (dd, J = 5.7, 3.7 Hz, 2H), 6.79 (dd, J = 5.5, 1 .1 Hz, 1 H), 2.52 (s, 3H), 2.44 (s, 3H). MS (ESI+) m/z: 322.1 [M + H]+. Mp: 308°C-31 1 0C.
Example B2-37 / PKE 360: Sodium 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)-thiazole-5-carboxylate (sodium salt of compound PKE 262): Method as for B2-34, using 4-methyl-2-(5-pyridin-3-yl-thiophen-2-yl)- thiazole-5-carboxylic acid B2-2 / PKE 262; yellow solid (1 00%). λ H NMR (DMSO) δ: 8.95 (d, J = 0.8 Hz, 1 H), 8.53 (dd, J = 4.7, 1 .5 Hz, 1 H), 8.1 1 (ddd, J = 8.0, 2.5, 1 .7 Hz, 1 H), 7.66 (d, J = 4.0 Hz, 1 H), 7.58 (d, J = 3.8 Hz, 1 H), 7.47 (ddd, J = 8.0, 4.9, 0.8 Hz, 1 H), 2.56 (s, 3H). MS (ESI+) m/z: 303.1 [M + H]+. Mp: >350°C.
Example B2-38 / PKE 348: Sodium 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-methyl-thiazole-5- carboxylate (sodium salt of compound PKE 361 ): Method as for B2-34, using 2-(5-benzo[b]thiophen-2- yl-thiophen-2-yl]-4-methyl-thiazole-5-carboxylic acid B2-6 / PKE 361 ; yellow solid (1 00%). 1 H NMR (DMSO) δ: 7.95 (dd, J = 6.6, 1 .9 Hz, 1 H), 7.83 (dd, J = 6.2, 1 .9 Hz, 1 H), 7.74 (s, 1 H), 7.42 (d, J = 3.9 Hz, 1 H), 7.39-7.33 (m, 3H), 2.55 (s, 3H). MS (ESI+) m/z: 358.1 [M + H]+. Mp: >300°C.
Amide thiazole series
Example B3-1 -1 / PKE 290: 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid: To a solution of 2-(5-bromo-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 -2 (50 mg, 0.1 5 mmol) dissolved in a 8/2 mixture of methanol-water (25 ml) was added lithium hydroxide monohydrate (25 mg, 0.60 mmol). The reaction mixture was heated at reflux for 2 hours. The solvent was evaporated in vacuo, water (25 ml) was added, and the solution extracted with CH2CI2 (3 x 1 5 ml). The aqueous phase was acidified to pH 1 with cone. HCI and extracted twice with EtOAc. The organic phases were collected, dried (Na2SO4), filtered and evaporated in vacuo to afford a white powder (66%). 1 H NMR (DMSO) δ: 7.64 (d, J = 4.0 Hz, 1 H), 7.32 (d, J = 4.0 Hz, 1 H), 2.58 (s, 3H). MS (ESI+) m/z: 304.0-306.0 [M + H]+. Mp: 242 °C-244°C. Example B3-1 -2: 2-(5-bromo-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid (2-morpholin4-yl- ethyl)-amide: To a solution of 2-(5-bromo-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid B3-1 -1 (1 .64 mmol, 500 mg) in acetone (1 0 ml), between -1 0/-1 50C, was added triethylamine (4.36 mmol, 0.6 ml), isobutylchloroformate (1 .89 mmol, 0.1 8 ml) followed by 2-aminoethyl morpholine (1 .64 mmol, 0.22 ml). The reaction mixture was stirred at room temperature. After completion of the reaction and concentration, the crude was purified by silica gel column chromatography (EtOAc-MeOH: 9/1 ) to give white crystals (46%).
Example B3-1 -3 / PKE 1 89.1 : 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid (2-morpholin- 4-yl-ethyl)-amide: To a mixture, degassed under argon, of thiophene-2-boronic acid (0.1 6 g, 1 .3 mmol), 2-(5-bromo-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid (2-morpholin4-yl-ethyl)-amide B3-1 -2 (0.47 g, 1 .1 mmol), and powdered CsF (0.38 g, 2.5 mmol) in 1 0 ml of DME was added Pd(PPh3)4 (44 mg, 0.04 mmol, 3 mol percent). Reaction mixture was degassed under argon and heated up to reflux for 1 8 h. Reaction mixture was then diluted with water and extracted with methylene chloride. Organic layer was dried and concentrated. Chromatography of the residue on silica gel (Et0Ac/Me0H/NH40H: 90/9/1 ) afforded a yellow solid (1 5%). 1 H NMR (DMSO) δ: 8.1 5 (t, J = 5.4 Hz, I H), 7.65 (d, J = 4.0 Hz, 1 H), 7.61 (dd, J = 5.0, 1 .0 Hz, 1 H), 7.46 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.35 (d, J = 4.0 Hz, 1 H), 7.1 2 (dd, J = 5.0, 3.6 Hz, 1 H), 3.55 (t, J = 4.5 Hz, 4H), 3.33 (br s, 2H), 2.55 (s, 3H), 2.43 (m, 6H). MS (ESI+) m/z: 420.1 [M + H]+. Mp: 228°C-230°C. Compound PKE 1 89.2 is the hydrochloride of compound PKE 1 89.1 .
Example B3-2 / PKE 500: 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-methyl-thiazole-5-carboxylic acid (2-morpholin4-yl-ethyl)-amide: Method as for B3-1 -3, using benzo[b]thiophene-2-boronic acid; pale yellow solid (20%). 1 H NMR (DMSO) δ: 8.1 2 (br s, 1 H), 7.96 (br s, I H), 7.81 (m, 2H), 7.70 (s, I H), 7.50 (br s, 1 H), 7.39 (m, 2H), 3.56 (m, 4H), 3.33 (m, 4H), 2.57 (s, 3H), 2.41 (m, 4H). MS (ESI+) m/z: 470.2 [M + H]+. Mp: 201 °C-204°C.
Example B3-3 / PKE 499: 4-methyl-2-(5-naphthalen-2-yl-thiophen-2-yl)-thiazole-5-carboxylic acid (2- morpholin-4-yl-ethyl)-amide: Method as for B3-1 -3, using naphthalene-2-boronic acid; yellow solid (59%). λ H NMR (CDCI3) δ: 8.1 0 (s, 1 H), 7.85 (m, 3H), 7.75 (d, J = 8.4 Hz, 1 H), 7.55 (d, J = 3.8 Hz, 1 H), 7.49 (m, 2H), 7.42 (d, J = 3.8 Hz, 1 H), 6.62 (br s, 1 H), 3.75 (m, 4H), 3.54 (q, J = 5.7 Hz, 2H), 2.73 (s, 3H), 2.61 (U = 5.7 Hz, 2H), 2.53 (m, 4H). MS (ESI+) m/z: 464.2 [M + H]+. Mp: 1 80°C-1 82 °C.
Example B3-4-1 : 2-Bromo-4-methyl-l ,3-thiazole-5-carboxylic acid (3-morpholin4-yl-propyl)-amide: Method as for B3-1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid and 3-morpholin-4-yl propylamine; yellow oil (82%). Example B3-4-2 / PKE 1 92.1 : 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid (3-morpholin- 4-yl-propyl)-amide: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and 2-bromo4-methyl- thiazole-5-carboxylic acid (3-morpholin4-yl-propyl)-amide B3-4-1 ; yellow solid (39%). 1 H NMR (DMSO) δ: 8.22 (br s, 1 H), 7.62 (d, J = 6.1 Hz, 1 H), 7.58 (dd, J = 5.1 , 1 .0 Hz, 1 H), 7.45 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.34 (d, J = 6.1 Hz, 1 H), 7.1 2 (dd, J = 5.1 , 3.6 Hz, 1 H), 3.55 (t, J = 4.6 Hz, 4H), 3.24 (q, J = 5.9 Hz, 2H), 2.53 (s, 3H), 2.34-2.27 (m, 6H), 1 .65 (m, 2 H). MS (ESI+) m/z: 434.2 [M + H]+. Mp: 1 030C-1 050C. Compound PKE 1 92.2 is the hydrochloride of compound PKE 1 92.1 .
Example B3-5-1 : 2-Bromo4-methyl-l ,3-thiazole-5-carboxylic acid (3-imidazol-l -yl-propyl)-amide : Method as for B3-1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid and 3-imidazol-l -yl propylamine; color- less oil (61 %).
Example B3-5-2 / PKE 195: 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid (3-imidazol-l -yl- propyl)-amide: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and 2-bromo4-methyl- thiazole-5-ca rboxylic acid (3-imidazol-l -yl-propyl)-amide B3-5-1 ; green solid (39%). 1 H NMR (DMSO) δ: 8.28 (m, 1 H), 7.61 (d, J = 3.8 Hz, 2H), 7.59 (d, J = 5.0 Hz, 1 H), 7.45 (d, J = 2.7 Hz, 1 H), 7.34 (d, J = 3.8 Hz, I H), 7.1 9 (s, 1 H), 7.1 2 (dd, J = 5.0, 3.8 Hz, I H), 6.88 (s, 1 H), 4.00 (t, J = 6.8 Hz, 2H), 3.1 9 (q, J = 6.6 Hz, 2H), 2.54 (s, 3H), 1 .96 (m, 2H). 1 3C NMR (DMSO) δ: 1 60.9, 1 59.1 , 1 54.6, 1 39.6, 1 37.3, 1 35.5, 1 34.1 , 1 29.0, 1 28.6, 1 28.4, 1 26.9, 1 25.5, 1 25.4, 1 25.0, 1 1 9.3, 43.7, 36.7, 30.6, 1 6.9. MS (ESI+) m/z: 41 5.1 [M + H]+. Mp: 1 26°C-1 28°C.
Example B3-6-1 : 2-Bromo-4-methyl-l ,3-thiazole-5-carboxylic acid ( 1 -ethyl-pyrrol id i n-2-yl methyl)-a m ide: Method as for B3-1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid and C-(I -ethyl-pyrrolidin-2-yl)- methylamine; pinkish oil (72%).
Example B3-6-2 / PKE 204: 2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid (1 -ethyl- pyrrolidin-2-ylmethyl)-amide: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and 2-bromo-4- methyl-thiazole-5-carboxylic acid ( 1 -ethyl-pyrrol id i n-2-y lmethyl)-a m ide B3-6-1 ; yellow solid (1 2%). 1 H NMR (DMSO) δ: 8.05 (t, J = 5.4 Hz, 1 H), 7.64 (d, J = 3.9 Hz, 1 H), 7.59 (d, J = 5.0 Hz, 1 H), 7.45 (d, J =
3.5 Hz, 1 H), 7.34 (d, J = 3.9 Hz, 1 H), 7.1 2 (dd, J = 5.0, 3.7 Hz, 1 H), 3.36 (m, 2H), 3.06 (m, 2H), 2.79 (m, 1 H), 2.53 (s, 3H), 2.29 (m, 1 H), 2.20 (m, 1 H), 1 .82 (m, 1 H), 2.63 (m, 3H), 1 .03 (t, J = 6.1 Hz, 3H). MS (ESI+) m/z: 41 8.2 [M + H]+. Mp: 66°C-68°C.
Example B3-7-1 : (2-Bromo4-methyl-l ,3-th i a zo l-5-y I )-[4-(4-f I uorophenyl)-piperazin-1 -yl]-methanone: Method as for B3-1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid and 1 -(4-fluorophenyl)- piperazine; white solid (1 5%). 1 H NMR (CDCI3) δ: 6.95-6.88 (m, 2H), 6.87-6.81 (m, 2H), 3.71 (m, 4H),
3.06 (t, J = 5.0 Hz, 4H), 2.40 (s, 3H). Example B3-7-2 / PKE 208: 2-[2,2']Bithiophenyl-5-yl4-methyl-thiazol-5-yl-[4-(4-fluorophenyl)-piperazin- l -yl]-methanone: Method as for B3-1 -3, using 2.2'-bithiophene-5-boronic acid and (2-bromo4-methyl- thiazole-5-yl)-[4-(4-fluoro-phenyl)-piperazin-l -yl]-methanone B3-7-1 ; yellow solid (3%). MS (ESI+) m/z: 470.0 [M + H]+. Mp: 1 25°C-1 26°C.
Example B3-8 / PKE 21 5: 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid isobutyl a mide: To a suspension of 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid B2-1 (307 mg, 1 .0 mmol) in dichloromethane (1 0ml) at room temperature are added oxalyl chloride (0.4 ml, 4.7 mmol) and N1N- dimethylformamide (0.4 ml). Reaction mixture is stirred at room temperature until complete dissolution (2h) and solvents are evaporated to dryness. Residue is dissolved back in dichloromethane (1 0ml) and isobutylamine (0.6 ml, 6.0 mmol) is added. Reaction mixture is stirred at room temperature for 1 2 h, then diluted with EtOAc and washed twice with water. Organic layers are dried and evaporated. Solid obtained is washed with diethyl ether and dried to afford a yellow solid (33%). 1 H NMR (CDCI3) δ: 7.41 (d, J = 3.9 Hz, 1 H), 7.25 (m, 2H), 7.1 3, (d, J = 3.9 Hz, 1 H), 7.04 (dd, J = 5.0, 3.7 Hz, 1 H), 5.82 (br s, 1 H), 3.26 (q, J = 6.1 Hz, 2H), 2.70 (s, 3H), 1 .90 (m, 1 H), 0.99 (d, J = 6.7 Hz, 6H). MS (ESI+) m/z: 363.1 [M + H]+. Mp: 1 31 0C-1 34°C.
Example B3-9-1 : 2-(5-bromo-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl amide: Method as for B3-1 -2, using ethylamine 2M solution in dioxane; white solid (55%). 1 H NMR (CDCI3) δ: 7.24 (d, J = 3.9 Hz, 1 H), 7.04 (d, J = 3.9 Hz, 1 H), 5.74 (br s, 1 H), 3.46 (qi, J = 7.1 Hz, 2H), 2.67 (s, 3H), 1 .25 (t, J = 7.1 Hz, 3H).
Example B3-9-2 / PKE 535: 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl amide: Method as for B3-1 -3, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-thiophen-2- yl)-4-methyl-thiazole-5-carboxylic acid ethyl amide B3-9-1 ; orange solid (72%). 1 H NMR (CDCI3) δ: 7.79- 7.72 (m, 2H), 7.45 (s, 1 H), 7.42 (d, J = 4.0 Hz, 1 H), 7.37-7.29 (m, 2H), 7.23 (d, J = 4.0 Hz, 1 H), 5.79 (br s, 1 H), 3.47 (qi, J = 7.1 Hz, 2H), 2.70 (s, 3H), 1 .26 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 385.2 [M + H]+. Mp: 214°C-21 6°C.
Example B3-1 0-1 : [2-(5-bromo-thiophen-2-yl)4-methyl-l ,3-thiazol-5-yl]-pyrrolidin-l -yl-methanone: Method as for B3-l -2, using pyrrolidine; yellow oil (51 %).
Example B3-1 0-2 / PKE 536: [2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazol-5-yl]-pyrrolidin- 1 -yl-methanone: Method as for B3-1 -3, using benzo[b]thiophene-2-boronic acid and [2-(5-bromo- thiophen-2-yl)4-methyl-thiazol-5-yl]-pyrrolidin-l -yl-methanone B3-1 0-1 ; yellow solid (28%). 1 H NMR (CDCI3) δ: 7.79 (dd, J = 7.0, 0.6 Hz, 1 H), 7.75 (dd, J = 6.3, 2.3 Hz, 1 H), 7.46 (s, 1 H), 7.43 (d, J = 3.9 Hz, 1 H), 7.36-7.29 (m, 2H), 7.24 (d, J = 4.0 Hz, 1 H), 3.58-3.50 (m, 4H), 2.56 (s, 3H), 1 .99-1 .93 (m, 4H). MS (ESI+) m/z: 41 1 .2 [M + H]+. Mp: 1 31 0C-1 33°C. Example B3-1 1 -1 : 2-bromo4-methyl-l ,3-thiazole-5-carboxylic acid amide: A suspension of 2-bromo-4- methyl-1 ,3-thiazole-5-carboxylic acid ethyl ester (0.60 g, 2.4 mmol) in a 30% aqueous ammonia solution (1 5 ml) was stirred at room temperature for 48 hours. Precipitate was filtered off, washed with water and dried to afford a white solid (94%). 1 H NMR (CDCI3) δ: 5.79 (br s, 2H), 2.68 (s, 3H). MS 5 (ESI+) m/z: 220.9-222.9 [M + H]+.
Example B3-1 1 -2 / PKE 543: 2-[2,2']bithiophenyl-5-yl4-methyl-thiazole-5-carboxylic acid amide
Figure imgf000064_0001
Method as for Bl -1 -1 , using 2-bromo-4-methyl-thiazole-5-carboxylic acid amide B3-1 1 -1 and 2,2'- bithiophenyl-5-boronic acid; green solid (65%). λ H NMR (CD3OD) δ: 7.56 (dd, J = 4.0, 1 .0 Hz, 1 H), 7.42 i o (d, J = 4.1 Hz, 1 H), 7.34 (dd, J = 2.6, 1 .0 Hz, 1 H), 7.23 (d, J = 3.9 Hz, 1 H), 7.08 (dd, J = 5.1 , 3.7 Hz, 1 H), 2.63 (s, 3H). MS (ESI+) m/z: 307.0 [M + H]+. Mp: 1 86°C-1 900C.
Alcohol thiazole series
Example B4-1 / PKE 213: (2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazol-5-yl)-methanol: To a suspension of lithium aluminium hydride (2.64 mmol, 1 00 mg) in dry TH F (1 0 ml) at 00C, was added 2-
15 [2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -2 (0.80 mmol, 268 mg). The reaction mixture was stirred at room temperature for 3 hours. After hydrolysis and concentration, the residue was purified by silica gel column chromatography (EtOAc) to give a yellow solid (1 9%). λ H NMR (CDCI3) δ: 7.33 (d, J = 3.9 Hz, 1 H), 7.25 (s, 1 H), 7.24 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.1 1 (d, J = 3.9 Hz, 1 H), 7.03 (dd, J = 5.0, 3.7 Hz, I H), 4.80 (s, 2H), 2.41 (s, 3H). MS (ESI+) m/z: 294.0 [M + H]+. Mp: 87°C-
20 900C.
Example B4-2 / PKE 530: [2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)4-methyl-thiazole-5-yl] -methanol: Method as for B4-1 , using 2-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Bl -1 2; yellow solid (45%). λ H NMR (DMSO) δ: 7.95 (d, J = 6.8 Hz, 1 H), 7.83 (d, J = 6.7 Hz, 1 H), 7.75 (s, 1 H), 7.44 (d, J = 3.8 Hz, 1 H), 7.38 (d, J = 2.7 Hz, 1 H), 7.33 (m, 2H), 5.57 (t, J = 5.4 Hz, 25 1 H), 4.62 (d, J = 5.1 Hz, 2H), 2.30 (s, 3H). MS (ESI+) m/z: 344.1 [M + H]+. Mp: 1 69°C-1 700C.
Ester thiazole series
Example B5-1 / PKE 205: 2-[2,2']Bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid (2-morpholin4-yl- ethyl)-ester: Method as for B3-8, using 2-[2,2']bithiophenyl-5-yl-4-methyl-thiazole-5-carboxylic acid B2-1 and 2-hydroxy-ethylmorpholine; yellow solid (1 9%). 1 H NMR (DMSO) δ: 7.77 (d, J = 4.0 Hz, I H), 7.61 (dd, J = 6.0, 0.9 Hz, 1 H), 7.48 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.37 (d, J = 4.0 Hz, 1 H), 7.1 3 (dd, J = 3.7, 6.0 Hz, 1 H), 4.34 (t, J = 5.8 Hz, 2H), 3.54 (t, J = 4.6 Hz, 4H), 2.63 (t, J = 5.8 Hz, 2H), 2.62 (s, 3H), 2.45 (m, 4H). MS (ESI+) m/z: 421 .1 [M + H]+. Mp: 84°C-86°C.
Central thiophen series
Example Cl -1 -1 : 4-methyl-2-(3-methyl-thiophen-2-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -M , using 3-methyl-thiophene-2-boronic acid; pale yellow solid (65%). 1 H NMR (CDCI3) δ: 7.34 (d, J = 5.0 Hz, 1 H), 6.93 (d, J = 5.0 Hz, 1 H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.52 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H).
Example Cl -1 -2: 2-(5-bromo-3-methyl-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -I -2, using 4-methyl-2-(3-methyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Cl - 1 -1 ; white solid (99%). 1 H NMR (CDCI3) δ: 7.1 1 (s, I H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.50 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H).
Example Cl -1 -3 / PKE 414: 4-methyl-2-(4-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-3-methyl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester CM -2 and thiophene-2-boronic acid; yellow solid (37%). 1 H NMR (CDCI3) δ: 7.26 (m, 2H), 7.02 (m, 2H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.49 (s, 3H), 1 .39 (t, J = 7.0 Hz, 3H). MS (ESI+) m/z: 350.1 [M + H]+. Mp: 1 63°C-1 66°C.
Example Cl -1 -4 / PKE 41 3: 2-(5-Benzo[b]thiophen-2-yl-3-methyl-thiophen-2-yl)4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-3-methyl-thiophen-2-yl)4-methyl- thiazole-5-carboxylic acid ethyl ester Cl -1 -2 and benzo[b]thiophene-2-boronic acid; yellow solid (41 %). 1 H NMR (CDCI3) δ: 7.80-7.74 (m, 2H), 7.48 (s, 1 H), 7.38-7.32 (m, 2H), 7.14 (s, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.77 (s, 3H), 2.53 (s, 3H), 1 .37 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 400.2 [M + H]+. Mp: 1 84°C- 1 86°C.
Example CM -5 / PKE 424: 4-Methyl-2-(3-methyl-5-p-tolyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-(5-bromo-3-methyl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester CM -2 and 4-methyl-benzene boronic acid; yellow solid (1 9%). 1 H NMR (CDCI3) δ: 7.53 (d, J = 7.8 Hz, 2 H), 7.20 (d, J = 7.8 Hz, 2H), 7.1 3 (s, 1 H), 4.35 (q, J = 7.1 Hz, 2 H), 2.77 (s, 3H), 2.53 (s, 3H), 2.37 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 358.2 [M + H]+. Mp: 1 84°C-1 86°C.
Example Cl -1 -6 / PKE 41 1 : 2-(5-Benzo[b]thiophen-2-yl-3-methyl-thiophen-2-yl)4-methyl-thiazole-5- carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-3-methyl-thiophen-2-yl)4-methyl- thiazole-5-carboxylic acid ethyl ester Cl -M; yellow solid (88%). 1 H NMR (DMSO) δ: 7.95 (d, J = 8.5 Hz, 1 H), 7.82 (d, J = 8.7 Hz, 1 H), 7.75 (s, 1 H), 7.41 (s, 1 H), 7.38-7.36 (m, 2H), 2.62 (s, 3H), 2.45 (s, 3H). MS (ESI+) m/z: 372.1 [M + H]+. Mp: 247 °C-249°C.
Example Cl -1 -7 / PKE 41 2: 4-Methyl-2-(4-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester Cl -1 -3; yellow solid (90%). λ H NMR (DMSO) δ: 7.59 (d, J = 5.0 Hz, 1 H), 7.42 (d, J = 2.8 Hz, 1 H), 7.30 (s, 1 H), 7.1 2 (dd, J = 4.9, 3.7 Hz, 1 H), 2.62 (s, 3H), 2.44 (s, 3H). MS (ESI+) m/z: 322.1 [M + H]+. Mp: 237 °C-240°C.
Example Cl -2-1 : 4-Methyl-2-(4-methyl-thiophen-2-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 4-methyl-thiophene-2-boronic acid; beige solid (98%). 1 H NMR (CDCI3) δ: 7.37 (d, J = 0.9 Hz, 1 H), 7.03 (s, 1 H), 4.32 (q, J = 7.1 Hz, 2H), 2.72 (s, 3H), 2.28 (s, 3H), 1 .37 (t, J = 7.1 Hz, 3H).
Example Cl -2-2: 2-(5-Bromo-4-methyl-thiophen-2-yl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -2, using 4-methyl-2-(4-methyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester Cl - 2-1 ; beige solid (68%). 1 H NMR (CDCI3) δ: 7.25 (s, I H), 4.32 (q, J = 7.1 Hz, 2H), 2.71 (s, 3H), 2.20 (s, 3H), 1 .37 (t, J = 7.1 Hz, 3H).
Example Cl -2-3: 4-Methyl-2-(3-methyl-[2,2']bithiophenyl-5-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo-4-methyl-thiophen-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Cl -2-2 and thiophene-2-boronic acid; yellow solid (28%). 1 H NMR (CDCI3) δ: 7.51 (m, I H), 7.36 (s, 1 H), 7.24 (d, J = 3.4 Hz, 1 H), 7.09 (dd, J = 3.8, 4.9 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2 H), 2.74 (s, 3H), 2.42 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H).
Example Cl -2-4 / PKE 399: 2-(5-Benzo[b]thiophen-2-yl4-methyl-thiophen-2-yl)4-methyl-thiazole-5- carboxylic acid ethyl ester: Method as for Al -2, using 2-(5-bromo4-methyl-thiophen-2-yl)4-methyl- thiazole-5-carboxylic acid ethyl ester Cl -2-2 and benzo[b]thiophene-2-boronic acid; yellow solid (1 9%). 1 H NMR (CDCI3) δ: 7.84-7.77 (m, 2H), 7.43 (s, 1 H), 7.40 (s, 1 H), 7.38-7.34 (m, 2H), 4.35 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.50 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 400.2 [M + H]+. Mp: 1 72 °C- 1 73°C.
Example Cl -2-5 / PKE 395: 4-Methyl-2-(3-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(3-methyl-[2,2']bithiophenyl-5-yl)-thiazole-5-carboxylic acid ethyl ester Cl -2-3; ochre solid (45%). 1 H NMR (DMSO) δ: 7.65 (m, 2H), 7.36 (s, I H), 7.1 7 (m, I H), 2.59 (s, 3H), 2.36 (s, 3H). MS (ESI+) m/z: 322.1 [M + H]+. Mp: 224°C-227 °C dec.
Example Cl -3-1 / PKE 256: 4-Methyl-2-thiophene-3-yl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester and thiophene-3-boronic acid; white solid (91 %). 1 H NMR (CD3OD) δ: 8.07 (dd, J = 2.5, 1 .7 Hz, I H), 7.54 (m, 2H), 4.30 (q, J = 7.1 Hz, 2H), 2.68 (s, 3H), 1 .35 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 254.0 [M + H]+. Mp: 41 °C-43°C.
Example Cl -3-2: 2-(2-Bromo-thiophene-3-yl)4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: 4-Methyl- 2-thiophene-3-yl-thiazole-5-carboxylic acid ethyl ester Cl -3-1 (840 mg, 3.30 mmol) and bromine (53 mg, 3.30 mmol) in glacial acetic acid (40 ml) were heated at 700C for 1 8 hours. Reaction mixture was concentrated. Residue was dissolved back in EtOAc (30 ml). Organic layers were washed twice with 1 0% aqueous K2CO3, 1 0% aqueous NaHSO3, and H2O. They were then concentrated, and dried under vacuum to give a pink solid (82 %).
Example Cl -3-3 / PKE 286: 2-[2,2']-Bithiophenyl-3-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(2-bromo-thiophene-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Cl - 3-2 and thiophene-2-boronic acid; pink solid (30%). 1 H NMR (CDCI3) δ: 7.61 (d, J = 5.5 Hz, I H), 7.47 (dd, J = 5.1 , 1 .1 Hz, 1 H), 7.34 (d, J = 5.4 Hz, 1 H), 7.28 (dd, J = 3.6, 1 .1 Hz, 1 H), 7.1 2 (dd, J = 5.1 , 3.6 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 1 .33 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 336.1 [M + H]+. Mp: 980C-I OO0C.
Example Cl -3-4/ PKE 323: 2-(5'-Acetyl-[2,2']-bithiophenyl-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 2-(2-bromo-thiophene-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester Cl -3-2 and 2-acetyl-thiophene-5-boronic acid; pale yellow solid (26%). 1 H NMR (CDCI3) δ: 7.64 (d, J = 3.7 Hz, 1 H), 7.53 (d, J = 5.2 Hz, 1 H), 7.40 (d, J = 5.0 Hz, 1 H), 7.34 (d, J = 3.8 Hz, 1 H), 4.32 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 2.58 (s, 3H), 1 .35 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 378.1 [M + H]+. Mp: 1 25 °C-1 27 °C.
Example Cl -4 / PKE 426: 4-Methyl-2-(4-phenyl-thiophen-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 4-phenylthiophene-2-boronic acid pinacol ester; pale yellow solid (23%). 1 H NMR (CDCI3) δ: 7.84 (d, J = 1 .4 Hz, 1 H), 7.62 (dd, J = 3.5, 1 .4 Hz, 2H), 7.55 (d, J = 1 .4 Hz, 1 H), 7.45- 7.40 (m, 2H), 7.36-7.33 (m, I H), 4.35 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 330.2 [M + H]+. Mp: 1 01 0C-1 04°C.
Central furan series
Example C2-1 -1 : 4-Methyl-2-furan-2-yl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using furane-2-boronic acid; grey solid (65%). λ H NMR (CDCI3) δ: 7.54 (d, J = 1 .2 Hz, 1 H), 7.09 (d, J = 3.4 Hz, 1 H), 6.55 (dd, J = 3.5, 1 .8 Hz, 1 H), 4.33 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .42 (t, J = 7.1 Hz, 3H).
Example C2-1 -2: 2-(5-Bromo-furan-2-yl)4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -2, using 4-methyl-2-furan-2-yl-thiazole-5-carboxylic acid ethyl ester C2-1 -1 ; pink solid (65%). 1 H NMR (CDCI3) δ: 7.04 (d, J = 3.5 Hz, 1 H), 6.48 (d, J = 3.5 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.74 (s, 3H), 1 .37 (t, J = 7.1 Hz, 3H).
Example C2-2 / PKE 523: 2-(5-Benzo[b]thiophen-2-yl-furan-2-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-(5-bromo-furan-2-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C2-1 -2 and benzo[b]thiophene-2-boronic acid; pale yellow solid (1 5%). 1 H NMR (CDCI3) δ: 7.83- 7.78 (m, 2H), 7.65 (s, 1 H), 7.38-7.34 (m, 2H), 7.21 (d, J = 3.7 Hz, 1 H), 6.77 (d, J = 3.6 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.78 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 370.1 [M + H]+. Mp: 1 52 °C- 1 54°C.
Example C2-3 / PKE 533: 2-(5-Benzo[b]thiophen-2-yl-furan-2-yl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-furan-2-yl)-4-methyl-thiazole-5-ca rboxylic acid ethyl ester C2-2; yellow solid (99%). λ H NMR (DMSO) δ: 1 3.44 (br s, 1 H), 8.00 (dd, J = 6.4, 3.2 Hz, 1 H), 7.90 (s, I H), 7.88 (d, J = 3.5 Hz, I H), 7.42-7.39 (m, 3H), 7.20 (d, J = 3.7 Hz, I H), 2.65 (s, 3H). MS (ESI+) m/z: 342.1 [M + H]+. Mp: 266°C-269°C.
Central thiazole series
Example C3-1 / PKE 450: 4,4'-Dimethyl-2'-phenyl-[2,5']bithiazolyl-5-carboxylic acid ethyl ester: Method as for Bl -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 4-methyl-2-phenyl-5- (4,4,5,5-tetramethyl-l ,3,2-dioxoborolan-2-yl)-l ,3-thiazole; pale yellow solid (54%). 1 H NMR (CDCI3) δ: 7.99-7.95 (m, 2H), 7.46-7.44 (m, 3H), 4.37 (q, J = 7.1 Hz, 2H), 2.77 (s, 3H), 2.76 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 345.2 [M + H]+. Mp: 143°C-145°C.
Example C3-2 / PKE 447: 4,4'-Dimethyl-2'-phenyl-[2,5']bithiazolyl-5-carboxylic acid: Method as for B2- 1 , using 4,4'-dimethyl-2'-phenyl-[2,5']bithiazolyl-5-carboxylic acid ethyl ester C3-1 ; white solid (75%). 1 H NMR (DMSO) δ: 7.98-7.95 (m, 2H), 7.52-7.50 (m, 3H), 2.67 (s, 3H), 2.63 (s, 3H). MS (ESI+) m/z: 31 7.1 [M + H]+. Mp: 244°C-247 °C.
Central phenyl series
Example C4-1 -1 : 2-(3-Hydroxy-phenyl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 3-hydroxyphenyl-boronic acid; white solid (49%). λ H NMR (CDCI3) δ: 7.48 (m, 1 H), 7.44 (dt, J = 7.7, 1 .5 Hz, 1 H), 7.29 (d, J = 7.9 Hz, 1 H), 6.94 (ddd, J = 8.1 , 2.5, 1 .1 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.76 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 264.0 [M + H]+.
Example C4-1 -2: 4-Methyl-2-(3-trifluoromethanesulfonyloxy-phenyl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Under argon at -300C, 2.6-lutidine (350 mg, 3.27 mmol) was added dropwise to a solution of 2- (3-hydroxy-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-1 -1 (500 mg, 1 .90 mmol) in dry methylene chloride. Then, at -300C, were added 4-/V,/V'-dimethylaminopyridine (1 00 mg, 0.82 mmol) and trifluoromethanesulfonic anhydride (0.5 ml, 1 .77 mmol). The mixture was stirred at room temperature for 1 6 h. After concentration in vacuo, the residue was purified by silica gel column chromatogra- phy (EtOAc/cyclohexane: 2/8) to give white needles (93%).
Example C4-1 -3 / PKE 386: 4-Methyl-2-(3-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(3-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-l -2 and thiophene-2-boronic acid; white solid (30%). 1 H NMR (CDCI3) δ: 8.20 (d, J = 1 .6 Hz, 1 H), 7.86 (d, J = 7.7 Hz, 1 H), 7.69 (d, J = 7.8 Hz, 1 H), 7.47 (d, J = 7.8 Hz, 1 H), 7.41 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.33 (dd, J = 5.1 , 0.9 Hz, 1 H), 7.1 1 (m, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.80 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 330.1 [M + H]+. Mp: 1 060C-1 080C.
Example C4-1 -4 / PKE 383: 2-(3-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(3-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-1 -2 and benzo[b]thiophene-2-boronic acid; white solid (65%). 1 H NMR (CDCI3) δ: 8.32 (s, 1 H), 7.91 (d, J = 7.8 Hz, 1 H), 7.85 (d, J = 7.7 Hz, 1 H), 7.83-7.78 (m, 2H), 7.66 (s, 1 H), 7.50 (t, J = 7.8 Hz, 1 H), 7.40-7.31 (m, 2H), 4.37 (q, J = 7.1 Hz, 2H), 2.82 (s, 3H), 1 .41 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 380.1 [M + H]+. Mp: 1 140C-I l 60C
Example C4-1 -5 / PKE 387: 2-(3-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(3-benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-M; pale yellow solid (93%). 1 H NMR (DMSO) δ: 8.31 (d, J = 1 .5 Hz, I H), 8.03 (s, I H), 8.00- 7.86 (m, 4H), 7.61 (t, J = 7.8 Hz, I H), 7.41 -7.37 (m, 2H), 2.70 (s, 3H). MS (ESI+) m/z: 352.1 [M + H]+. Mp: 21 9°C-221 °C.
Example C4-2-1 / PKE 445: 2-(4-Hydroxy-phenyl)4-methyl-thiazole-5-carboxylic acid ethyl este: Method as for Al -1 -2, using 2-bromo-4-methyl-thiazole-5-carboxylic acid ethyl ester and 4-hydroxyphenyl-boronic acid; white solid (43%). 1 H NMR (CDCI3) δ: 7.81 (td, J = 8.8, 2.2 Hz, 2H), 6.84 (td, J = 8.8, 2.2 Hz, 2 H), 6.60 (br s, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.75 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 264.0 [M + H]+. Mp: 1 8O0C-1 83°C.
Example C4-2-2: 4-Methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for C4-l -2, using 2-(4-hydroxy-phenyl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4- 2-1 ; white solid (81 %).
Example C4-2-3 / PKE 384: 4-Methyl-2-(4-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-2 and thiophene-2-boronic acid; white solid (57%). 1 H NMR (CDCI3) δ:7.97 (d, J = 8.5 Hz, 2H), 7.69 (d, J = 8.5 Hz, 2H), 7.41 (dd, J = 3.6, 1 .0 Hz, 1 H), 7.34 (dd, J = 5.1 , 0.9 Hz, 1 H), 7.1 1 (dd, J = 5.0, 3.7 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.79 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 330.1 [M + H]+. Mp: 97 °C-99°C.
Example C4-2-4 / PKE 442: 2-(4-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-2 and benzo[b]thiophene-2-boronic acid; beige solid (71 %). 1 H NMR (CDCI3) δ: 8.00 (dd, J = 6.7, 1 .8 Hz, 2H), 7.84-7.77 (m, 2H), 7.76 (dd, J = 6.7, 1 .8 Hz, 2H), 7.63 (s, 1 H), 7.38-7.29 (m, 2H), 4.35 (q, J = 7.1 Hz, 2H), 2.78 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 380.2 [M + H]+. Mp: 1 89°C-1 91 °C.
Example C4-2-5 / PKE 408: 2-Biphenyl4-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 4-biphenyl boronic acid; white solid (45%). 1 H NMR (CDCI3) δ: 8.04 (dd, J = 6.6, 1 .9 Hz, 2H), 7.69 (dd, J = 6.6, 1 .9 Hz, 2H), 7.66-7.63 (m, 2H), 7.50-7.36 (m, 3H), 4.37 (q, J = 7.1 Hz, 2H), 2.80 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 324.2 [M + H]+. Mp: 1 060C-1 080C.
Example C4-2-6 / PKE 457: 4-Methyl-2-(4-naphthalen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using 4-methyl-2-(4-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-2 and naphthalene-2-boronic acid; white solid (59%). 1 H NMR (CDCI3) δ: 8.07-8.04 (m, 3H), 7.93-7.87 (m, 3H), 7.83-7.76 (m, 3H), 7.56-7.49 (m, 2H), 4.38 (q, J = 7.1 Hz, 2H), 2.82 (s, 3H), 1 .41 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 374.3 [M + H]+. Mp: 1 090C-H l 0C.
Example C4-2-7 / PKE 400: 4-Methyl-2-(4-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-3; yellow solid (22%). MS (ESI+) m/z: 302.1 [M + H]+. Mp: 21 1 °C-21 3°C.
Example C4-2-8 / PKE 458: 2-(4-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(4-benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic ethyl ester C4-2-4; yellow solid (96%). λ H NMR (DMSO) δ: 8.06 (d, J = 8.5 Hz, 1 H), 8.03 (d, J = 1 0.3 Hz, 2H), 7.99 (dd, J = 8.0, 2.3 Hz, 1 H), 7.91 (d, J = 8.5 Hz, 1 H), 7.89 (t, J= 1 0.1 Hz, 2H), 7.39 (m, J = 3.2, 2.8 Hz, 2H), 2.68 (s, 3H). MS (ESI+) m/z: 352.1 [M + H]+. Mp: 250°C-255°C.
Example C4-2-9 / PKE 432: 2-Biphenyl4-yl-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-Biphenyl-4-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-2-5; white solid (82%). 1 H NMR (DMSO) δ: 8.06 (d, J = 8.3 Hz, 2H), 7.82 (d, J = 8.3 Hz, 2H), 7.75 (d, J = 7.4 Hz, 2H), 7.50 (t, J = 7.2 Hz, 2H), 7.44-7.39 (m, 1 H), 2.69 (s, 3H). MS (ESI+) m/z: 296.1 [M + H]+. Mp: 237 °C-239°C. Example C4-2-1 0 / PKE 460: 4-Methyl-2-(4-naphthalen-2-yl-phenyl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4-naphthalen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-2-6; yellow solid (86%). 1 H NMR (DMSO) δ: 8.33 (s, 1 H), 8.1 1 (d, J = 8.4 Hz, 2H), 8.05-7.89 (m, 6H), 7.55 (m, J = 3.8 Hz, 2H), 2.70 (s, 3H). MS (ESI+) m/z: 346.2 [M + H]+. Mp: 250°C-253°C.
Example C4-3-1 : 2-(2-Hydroxy-phenyl)-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Al -1 -2, using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 2-hydroxyphenyl-boronic acid; beige solid. λ H NMR (CDCI3) δ: 7.70-7.63 (m, 1 H), 7.38-7.27 (m, 1 H), 7.08 (d, J = 8.3 Hz, 1 H), 6.91 (dt, J = 8.3, 1 .1 Hz, 1 H), 4.34 (q, J = 7.1 Hz, 2H), 2.78 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H).
Example C4-3-2: 4-Methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for C4-1 -2, using 2-(2-hydroxy-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-3-1 ; pale yellow oil (27% for two steps). 1 H NMR (CDCI3) δ: 8.06 (dd, J = 5.7, 2.0 Hz, I H), 7.57-7.44 (m, 3H), 4.37 (q, J = 7.1 Hz, 2H), 2.81 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H).
Example C4-3-3 / PKE 409: 2-(2-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-2 and benzo[b]thiophene-2-boronic acid; light pink solid (94%). 1 H NMR (CDCI3) δ: 7.90-7.85 (m, 1 H), 7.76 (ddd, J = 6.7, 2.5, 1 .0 Hz, 2H), 7.60-7.56 (m, 1 H), 7.52-7.48 (m, 2H), 7.37- 7.32 (m, 2H), 7.23 (dd, J = 1 0.2, 0.5 Hz, 1 H), 4.23 (q, J = 7.1 Hz, 2H), 2.73 (s, 3H), 1 .26 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 380.2 [M + H]+. Mp: 1 1 5°C-1 1 9°C.
Example C4-3-4 / PKE 434: 2-(2-Benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(2-benzo[b]thiophen-2-yl-phenyl)-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-3-3; white solid (67%). 1 H NMR (DMSO) δ: 7.96 (dd, J = 6.2, 1 .0 Hz, 2H), 7.84 (dd, J = 6.8, 1 .9 Hz, I H), 7.59 (m, 3H), 7.45 (s, I H), 7.39-7.35 (m, 2H), 2.58 (s, 3H). MS (ESI+) m/z: 352.2 [M + H]+. Mp: 229°C-232 °C.
Example C4-3-5 / PKE 41 0: 4-Methyl-2-(2-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-2 and thiophene-2-boronic acid; beige solid (88%). 1 H NMR (CDCI3) δ: 7.92 (m, I H), 7.50-7.44 (m, 3H), 7.37 (dd, J = 5.1 , 1 .2 Hz, 1 H), 7.03 (dd, J = 5.1 , 3.5 Hz, 1 H), 6.96 (dd, J = 3.5, 1 .2 Hz, 1 H), 4.28 (q, J = 7.1 Hz, 2H), 2.73 (s, 3H), 1 .32 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 330.1 [M + H]+. Mp: 1 000C-1 040C.
Example C4-3-6 / PKE 435: 4-Methyl-2-(2-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid: thod as for B2-1 , using 4-methyl-2-(2-thiophen-2-yl-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-5; white solid (55%). λ H NMR (DMSO) δ: 7.98 (dd, J = 5.0, 2.0 Hz, 1 H), 7.68 (dd, J = 5.0, 1 .1 Hz, 1 H), 7.58-7.52 (m, 3H), 7.1 3 (dd, J = 5.0, 3.5 Hz, 1 H), 7.09 (dd, J = 3.5, 1 .1 Hz, 1 H), 2.62 (s, 3H). MS (ESI+) m/z: 302.1 [M + H]+. Mp: 21 2 °C-214°C.
Example C4-3-7 / PKE 433: 4-Methyl-2-(4 '-methyl-biphenyl-2-yl)-thiazole-5-carboxylic acid ethyl ester: Method as for Al -2, using 4-methyl-2-(2-trifluoromethanesulfonyloxy-phenyl)-thiazole-5-carboxylic acid ethyl ester C4-3-2 and 4-tolyl boronic acid; white solid (82%). λ H NMR (CDCI3) δ: 7.99 (dd, J = 6.7, 1 .9 Hz, 1 H), 7.50-7.41 (m, 2H), 7.34 (dd, J = 6.5, 2.0 Hz, 1 H), 7.20-7.14 (m, 4H), 4.26 (q, J = 7.1 Hz, 2H), 2.71 (s, 3H), 2.40 (s, 3H), 1 .30 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 338.2 [M + H]+. Mp: 72 °C-76°C.
Example C4-3-8 / PKE 436: 4-Methyl-2-(4 '-methyl-biphenyl-2-yl)-thiazole-5-carboxylic acid: Method as for B2-1 , using 4-methyl-2-(4 '-methyl-biphenyl-2-yl)-thiazole-5-carboxylic acid ethyl ester C4-3-7; white solid (95%). 1 H NMR (DMSO) δ: 8.01 (dd, J = 7.6, 1 .5 Hz, 1 H), 7.53 (dq, J = 7.3, 1 .8 Hz, 2H), 7.34 (dd, J = 7.1 , 1 .5 Hz, 1 H), 7.24 (d, J = 7.9 Hz, 2H), 7.1 5 (d, J = 8.1 Hz, 2H), 2.60 (s, 3H), 2.36 (s, 3H). MS (ESI+) m/z: 31 0.1 [M + H]+ Mp: 1 85°C-1 90°C.
Example C4-4-1 : 2-(6-Hydroxy-pyridin-3-yl)4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using 2-bromo4-methyl-thiazole-5-carboxylic acid ethyl ester and 2-hydroxypyridine-5-boronic acid pinacol ester; brown solid (55%). 1 H NMR (DMSO) δ: 9.85 (br s, 1 H), 8.1 3 (d, J = 2.6 Hz, I H), 7.95 (dd, J = 9.6, 2.7 Hz, 1 H), 6.45 (d, J = 9.6 Hz, 1 H), 4.27 (q, J = 7.1 Hz, 2H), 2.63 (s, 3H), 1 .29 (t, J = 7.1 Hz, 3H).
Example C4-4-2: 4-Methyl-2-(6-trifluoromethanesulfonyloxy-pyridin-3-yl)-l ,3-thiazole-5-carboxylic acid ethyl ester: Method as for C4-1 -2, using 2-(6-hydroxy-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4^-l ; white solid (14%). λ H NMR (CDCI3) δ: 8.93 (d, J = 2.4 Hz, 1 H), 8.46 (td, J = 8.5, 1 .5 Hz, 1 H), 7.27 (d, J = 9.4 Hz, 1 H), 4.37 (q, J = 7.1 Hz, 2H), 2.79 (s, 3H), 1 .40 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 397.1 [M + H]+
Example C4-4-3 / PKE 437: 2-(6-Benzo[b]thiophen-2-yl-pyridin-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 4-methyl-2-(6- trifluoromethanesulfonyloxy-pyridin-3-yl)-thiazole-5-carboxylic acid ethyl ester C4-4-2; yellow solid (36%). 1 H NMR (CDCI3) δ: 9.1 5 (d, J = 1 .6 Hz, I H), 8.32 (dd, J = 8.4, 2.2 Hz, I H), 7.92 (s, I H), 7.90-7.82 (m, 3H), 7.41 -7.35 (m, 2H), 4.38 (q, J = 7.1 Hz, 2H), 2.81 (s, 3H), 1 .41 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 381 .3 [M + H]+. Mp: 1 88°C-1 90°C.
Example C4-4-4 / PKE 441 : 2-(6-Benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(6-benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4-4-3; yellow solid (90%). 1 H NMR (DMSO) δ: 9.1 3 (d, J = 1 .8 Hz, I H), 8.39 (dd, J = 8.4, 2.2 Hz, 1 H), 8.27 (s, 1 H), 8.20 (d, J = 8.4 Hz, 1 H), 7.97 (dd, J = 6.0, 3.1 Hz, 1 H), 7.88 (dd, J = 6.0, 3.1 Hz, 1 H), 7.38 (m, 2H), 2.67 (s, 3H). MS (ESI+) m/z: 353.2 [M + H]+. Mp: 273°C-275°C.
Example C4-5-1 : 5-Bromo-thionicotinamide: o,o'-Diethyl-dithiophosphate (3.3 ml, 21 .4 mmol) is added to a mixture of 5-bromonicotinonitrile (730 mg, 4.3 mmol) in pyridine (1 5 ml) and water (3 ml). Reac- tion mixture is stirred at 900C in a sealed tube for 72 h. Solvents are evaporated to dryness. Residue is dissolved back in EtOAc, and washed three times with water. Organic layer is dried (Na2SO4), filtered and solvents are evaporated to give a yellow solid. 1 H NMR (DMSO) δ: 1 0.1 8 (br s, 1 H), 9.78 (br s, I H), 8.96 (d, J = 1 .5 Hz, 1 H), 8.79 (d, J = 1 .6 Hz, 1 H), 8.37 (t, J = 1 .6 Hz, 1 H).
Example C4-5-2 / PKE 459: 2-(5-Bromo-pyridin-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: To a mixture of 5-bromo-thionicotinamide C4-5-1 (1 00 mg, 0.46 mmol) in absolute ethanol (3 ml) is added ethyl-2-chloroacetoacetate (0.064 ml, 0.46 mmol). Reaction mixture is heated up to 1 800C in a microwave apparatus for 1 5 min under pressure (20 bars). Solvents are evaporated to dryness. Residue is purified by silica gel column chromatography (cyclohexane/EtOAc 95/5) to give a white solid (46%). 1 H NMR (CDCI3) δ: 9.03 (d, J = 1 .8 Hz, 1 H), 8.73 (d, J = 2.1 Hz, 1 H), 8.43 (t, J = 2.0 Hz, 1 H), 4.36 (q, J = 7.1 Hz, 2H), 2.78 (s, 3H), 1 .38 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 327.0-329.0 [M + H]+. Mp: 1 09°C- 1 1 1 0C.
Example C4-5-3 / PKE 471 : 2-(5-Benzo[b]thiophen-2-yl-pyridin-3-yl)-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo-pyridin-3-yl)-4- methyl-thiazole-5-carboxylic acid ethyl ester C4-5-2; white solid (48%). 1 H NMR (CDCI3) δ: 9.07 (d, J = 2.0 Hz, 1 H), 9.01 (d, J = 2.2 Hz, 1 H), 8.51 (t, J = 2.1 Hz, 1 H), 7.87-7.80 (m, 2H), 7.70 (s, 1 H), 7.41 - 7.33 (m, 2H), 4.37 (q, J = 7.1 Hz, 2H), 2.81 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 381 .1 [M + H]+. Mp: 1 59°C-1 61 °C.
Example C4-5-4 / PKE 473: 2-(5-Benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-(5-benzo[b]thiophen-2-yl-pyridin-3-yl)4-methyl-thiazole-5-carboxylic acid ethyl ester C4-5-3; yellow solid (1 00%). 1 H NMR (DMSO) δ: 9.1 2 (d, J = 1 .9 Hz, 2H), 8.60 (t, J = 2.0 Hz, 1 H), 8.20 (s, 1 H), 8.02 (m, 1 H), 7.88 (m, 1 H), 7.43-7.40 (m, 2H), 2.71 (s, 3H). MS (ESI+) m/z: 353.1 [M + H]+. Mp: 280°C-283°C.
Example C4-6-1 / PKE 444: 2-Benzo[b]thiophen-2-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester: Method as for Bl -1 -1 , using benzo[b]thiophene-2-boronic acid and 2-bromo4-methyl-thiazole-5- carboxylic acid ethyl ester; beige solid (44%). λ H NMR (CDCI3) δ: 7.82 (m, 3H), 7.39 (t, J = 3.8 Hz, 2H), 4.35 (q, J = 7.1 Hz, 2H), 2.77 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 304.1 [M + H]+. Mp: 1 000C-1 030C. Example C4-6-2 / PKE 443: 2-Benzo[b]thiophen-2-yl-4-methyl-thiazole-5-carboxylic acid: Method as for B2-1 , using 2-benzo[b]thiophen-2-yl-4-methyl-thiazole-5-carboxylic acid ethyl ester C4-6-1 / PKE 444; green solid (84%). 1 H NMR (DMSO) δ: 8.1 8 (s, I H), 8.03 (dd, J = 6.5, 2.6 Hz, I H), 7.94 (dd, J = 7.0, 1 .6 Hz, 1 H), 7.47-7.43 (m, 2H), 2.65 (s, 3H). MS (ESI+) m/z: 276.0 [M + H]+. Mp: 262 °C-263°C.
Disruption of the conjugation series
Example Dl -I / PKE 467: 4-Methyl-2-[5-(thiophene-2-carbonyl)-thiophen-2-yl]-thiazole-5-carboxylic acid ethyl ester: To a mixture of thiophene-2-carboxylic acid chloride (0.1 1 ml, 1 .0 mmol) and anhydrous ferric chloride (1 65 mg, 2.0 mmol) in dry dichloromethane (25 ml) was added 4-methyl-2-thiophen-2-yl- thiazole-5-carboxylic acid ethyl ester Bl -1 -1 (253 mg, 1 .0 mmol). Reaction mixture was heated at reflux for 1 8 h, then concentrated in vacuo. The residue was purified by silica gel column chromatography (cyclohexane/EtOAc: 8/2) to afford a yellow solid (30Zo)- 1 H NMR (CDCI3) δ: 7.95 (dd, J = 3.7, 0.8 Hz, 1 H), 7.86 (d, J = 4.0 Hz, 1 H), 7.74 (m, 1 H), 7.59 (d, J = 4.0 Hz, 1 H), 7.20 (dd, J = 4.7, 3.9 Hz, 1 H), 4.37 (q, J = 7.1 Hz, 2 H), 2.77 (s, 3H), 1 .39 (t, J = 7.1 Hz, 3H). 1 3C NMR (CDCI3) δ: 1 78.2, 1 61 .9, 1 61 .8, 1 61 .2, 144.4, 143.1 , 142.6, 1 34.1 , 1 33.5, 1 33.4, 1 28.2, 1 27.6, 1 22.7, 61 .5, 1 7.4, 14.3. MS (ESI+) m/z: 364.1 [M + H]+. Mp: 1 1 1 0C-1 1 3°C.
Example D2-1 -1 : 2-[(5-Bromo-thiophene-2-carbonyl)-amino]-4-methyl-l ,3-thiazole-5-carboxylic acid ethyl ester: A solution of ethyl-2-amino4-methyl-thiazole-5-carboxylate (430 mg, 2.3 mmol), 5-bromo-2- thiophene carboxylic acid (400 mg, 1 .9 mmol), 1 -hydroxybenzotriazole hydrate (320 mg, 2.3 mmol) and l -(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (445 mg, 2.3 mmol) in N1N- dimethylformamide (20 ml) was heated up to 600C in a sealed tube for 1 8 h. After return to room temperature, a precipitate appeared. Solid was filtered off, washed with saturated NH4CI solution, saturated NaHCO3 solution and dried under vacuum to give a white solid (83%). 1 H NMR (DMSO) δ: 7.70 (d, J = 3.5 Hz, 1 H), 7.28 (d, J = 4.0 Hz, 1 H), 4.20 (q, J = 7.1 Hz, 2H), 2.51 (s, 3H), 1 .27 (t, J = 7.1 Hz, 3H).
Example D2-1 -2 / PKE 496: 2-[(5-Benzo[b]thiophen-2-yl-thiophene-2-carbonyl)-amino]4-methyl- thiazole-5-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 2- [(S-bromo-thiophene^-carbonylJ-aminoH-methyl-thiazole-S-carboxylic acid ethyl ester D2-1 -1 ; yellow solid (1 00%). ] H NMR (DMSO) δ: 7.95 (d, J = 7.2 Hz, 1 H), 7.85 (d, J = 6.7 Hz, 1 H), 7.68 (s, 1 H), 7.49 (d, J = 3.8 Hz, 1 H), 7.39-7.34 (m, 3H), 4.1 5 (q, J = 7.1 Hz, 2H), 2.46 (s, 3H), 1 .26 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 429.1 [M + H]+. Mp: >300°C.
Example D2-2 / PKE 497: 2-[(5-Benzo[b]thiophen-2-yl-thiophene-2-carbonyl)-methyl-amino]4-methyl- thiazole-5-carboxylic acid ethyl ester: To a suspension of 2-[(5-benzo[b]thiophen-2-yl-thiophene-2- carbonyl)-amino]4-methyl-thiazole-5<arboxylic acid ethyl ester D2-1 -2 (1 00 mg, 0.23 mmol) in N1N- dimethylformamide (5 ml) under argon at room temperature was added sodium hydride (60% dispersion in mineral oil, 1 3 mg, 0.32 mmol). Reaction mixture was stirred at room temperature for 20 minutes, and methyl iodide (0.044 ml, 0.71 mmol) was then added. Reaction mixture was heated up to 600C for 3 h. Saturated aqueous NH4CI was added, precipitate was filtered off, washed with saturated NaHCO3 solution, and dried under vacuum to give a yellow solid (73%). 1 H NMR (DMSO) δ: 7.98 (dd, J = 6.4, 2.4 Hz, 1 H), 7.87 (d, J = 2.5 Hz, 1 H), 7.84 (s, 1 H), 7.82 (d, J = 3.8 Hz, 1 H), 7.51 (d, J = 4.0 Hz, 1 H), 7.44-7.36 (m, 2H), 4.28 (q, J = 7.0 Hz, 2H), 3.80 (s, 3H), 2.69 (s, 3H), 1 .30 (t, J = 7.0 Hz, 3H). MS (ESI+) m/z: 443.1 [M + H]+. Mp: 234°C-238°C.
Example D2-3 / PKE 498: 2-[(5-Benzo[b]thiophen-2-yl-thiophene-2-carbonyl)-methyl-amino]4-methyl- thiazole-5-carboxylic acid: Method as for B2-1 , using 2-[(5-benzo[b]thiophen-2-yl-thiophene-2-carbonyl)- methyl-amino]-4-methyl-thiazole-5-carboxylic acid ethyl ester D2-2; yellow solid (75%). 1 H NMR (DMSO) δ: 7.97 (dd, J = 6.4, 2.6 Hz, 1 H), 7.86 (m, 1 H), 7.83 (s, 1 H), 7.81 (d, J = 3.9 Hz, 1 H), 7.50 (d, J = 3.9 Hz, I H), 7.42-7.38 (m, 2 H), 3.78 (s, 3H), 2.67 (s, 3H). MS (ESI+) m/z: 41 5.0 [M + H]+. Mp: 268°C- 272 °C.
Example D2-4 / PKE 469: 4-Methyl-2-[(thiophene-2-carbonyl)-amino]-thiazole-5-carboxylic acid ethyl ester: Method as for D2-1 -1 , using 2-thiophene carboxylic acid and ethyl-2-amino4-methyl-thiazole-5- carboxylate; pink solid (50%). λ H NMR (DMSO) δ: 1 3.1 0 (br s, 1 H), 8.24 (br s, 1 H), 7.99 (d, J = 4.7 Hz, 1 H), 7.25 (dd, J = 4.9, 3.9 Hz, 1 H), 4.25 (q, J = 7.1 Hz, 2H), 2.58 (s, 3H), 1 .28 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 297.1 [M + H]+. Mp: 245°C-247 °C.
Thiazole replacement
Example El -I / PKE 353: 4-methyl-[2,2',5',2"]terthiophene-5-carbaldehyde
Figure imgf000075_0001
Method as for Al -1 -2, using 5-formyl-4-methylthiophene-2-boronic acid and 5-bromo-2,2'bithiophene; yellow solid (21 %). 1 H NMR (CDCI3) δ 9.98 (s, I H), 7.27-7.20 (m, 3H), 7.1 1 (d, J = 3.8 Hz, I H), 7.05- 7.01 (m, 2H), 2.56 (d, J = 3.5 Hz, 3H). MS (ESI+) m/z: 291 .0 [M + H]+. Mp: 92 °C-94°C.
Example E2-1 -1 : 5-Bromo-[ l ,3,4]thiadiazole-2-carboxylic acid ethyl ester: Method as for Al -1 -1 , using 5- amino-[ l ,3,4]thiadiazole-2-carboxylic acid ethyl ester; white solid (82%). 1 H NMR (CDCI3) δ 4.53 (q, J = 7.2 Hz, 2H), 1 .46 (t, J = 7.2 Hz, 3H). Example E2-1-2 / PKE 454: 5-[2,2']Bithiophenyl-5-yl-[l,3,4]thiadiazole-2-carboxylic acid ethyl ester: Method as for Bl-1-1, using 2,2'-bithiophene-5-boronic acid and 5-Bromo-[l,3,4]thiadiazole-2-carboxylic acid ethyl ester E2-1-1; yellow solid (25%).1H NMR (CDCI3) δ 7.56 (d, J =4.0 Hz, IH), 7.34-7.31 (m, 2H), 7.21 (d, J = 4.0 Hz, IH), 7.07 (dd, J = 5.1, 3.8 Hz, 1 H), 4.53 (q, J = 7.2 Hz, 2H), 1.47 (t, J = 7.2 Hz, 3H). MS (ESI+) m/z: 323.1 [M + H]+. Mp: 145°C-147°C.
Example E2-2/ PKE 455: 2-[2,2']Bithiophenyl-5-yl-[l,3,4]thiadiazole: Method as for Bl-1-1, using 2,2'- bithiophene-5-boronic acid and 5-Bromo-[l,3,4]thiadiazole-2-carboxylic acid ethyl ester E2-1-1; grey flakes (36%).1H NMR (CDCI3) δ 9.03 (s, 1 H), 7.50 (d, J = 3.7 Hz, 1 H), 7.32-7.29 (m, 2H), 7.19 (d, J = 4.0 Hz, 1 H), 7.07 (dd, J = 5.0, 3.7 Hz, 1 H). MS (ESI+) m/z: 251.0 [M + H]+. Mp: 127°C-129°C.
Example E2-3-1: 2-Bromo-5-methyl-[l,3,4]thiadiazole: Method as for Al -1-1, using 5-methyl- [1 ,3,4]thiadiazol-2-ylamine; white solid (48%).1H NMR (CDCI3) δ 2.78 (s, 3H).
Example E2-3-2/ PKE 464: 2-[2,2']Bithiophenyl-5-yl-5-methyl-[l,3,4]thiadiazole: Method as for Bl-1-1, using 2,2'-bithiophene-5-boronic acid and 2-bromo-5-methyl-[l,3,4]thiadiazole E2-3-1; greenish flakes (50%).1H NMR (CDCI3) δ 7.39 (d, J = 2.8 Hz, IH), 7.28 (ddd, J = 5.1, 3.7, 1.3 Hz, 2H), 7.15 (d, J = 2.8 Hz, IH), 7.05 (dd, J = 3.7, 2.6 Hz, IH), 2.80 (s, 3H). MS (ESI+) m/z: 265.0 [M + H]+. Mp: 145°C- 147°C.
Example E2-4-1: 2-Bromo-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Al -1-1, using 2-amino-5- trifluoromethyl-[l,3,4]thiadiazole; yellow oil (43%).13C NMR (CDCI3) δ 142.7, 120.2, 116.6.
Example E2-4-2/ PKE 504: 2-[2,2']Bithiophenyl-5-yl-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Bl-1-1, using 2,2'-bithiophene-5-boronic acid and 2-bromo-5-trifluoromethyl-[l,3,4]thiadiazole E2-4-1; greenish solid (20%).1H NMR (CDCI3) δ 7.56 (d, J = 3.7 Hz, 1 H), 7.36-7.32 (m, 2H), 7.22 (d, J = 4.0 Hz, 1 H), 7.08 (dd, J = 5.0, 3.6 Hz, 1 H). MS (ESI+) m/z: 319.1 [M + H]+. Mp: 175°C-177°C.
Example E2-4-3: 2-Thiophen-2-yl-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Bl-1-1, using thio- phene-2-boronic acid and 2-bromo-5-trifluoromethyl-[l,3,4]thiadiazole E2-4-1; yellow solid (44%). 1H NMR (CDCI3) δ 7.66 (dd, J = 3.8, 1.1 Hz, 1 H), 7.61 (dd, J = 5.1 , 1.1 Hz, 1 H), 7.18 (dd, J = 5.1 , 3.8 Hz, 1 H). MS (ESI+) m/z: 236.9 [M + H]+.
Example E2-4-4: 2-(5-Bromo-thiophen-2-yl)-5-trifluoromethyl-[l,3,4]thiadiazole: Method as for Bl-1-2, using 2-thiophen-2-yl-5-trifluoromethyl-[l,3,4]thiadiazole E2-4-3; white solid (60%).1H NMR (CDCI3) δ 7.39 (d, J = 4.0 Hz, 1 H), 7.15 (d, J = 4.0 Hz, 1 H).
Example E2-4-5 / PKE 508: 2-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-5-trifluoromethyl- [l,3,4]thiadiazole: Method as for Bl-2, using benzo[b]thiophene-2-boronic acid and 2-(5-bromo- thiophen-2-yl)-5-trifluoromethyl-[ l ,3,4]thiadiazole E2-4-4; greenish solid (45%). 1 H NMR (CDCI3) δ 7.81 (dt, J = 6.6, 3.9 Hz, 2H), 7.60 (d, J = 4.0 Hz, 1 H), 7.57 (s, 1 H), 7.38 (dt, J = 3.8, 2.0 Hz, 2H), 7.33 (d, J = 3.9 Hz, 1 H). MS (ESI+) m/z: 369.0 [M + H]+. Mp: 255°C-260°C.
Example E3-1 -1 : 5-Bromo-2-methyl-benzoic acid ethyl ester: A solution of 5-bromo-2-methylbenzoic acid (200 mg, 0.93 mmol) and concentrated sulphuric acid (0.20 ml) in absolute ethanol (1 0 ml) was heated up to reflux for 1 5 h. Solvents were evaporated. Residue was dissolved back in EtOAc and washed with NaHCO3 solution and brine. Organic layers were dried (MgS04), and concentrated under vacuum to give a yellow oil (97%). 1 H NMR (CDCI3) δ 8.03 (d, J = 2.2 Hz, 1 H), 7.49 (dd, J = 8.3, 2.2 Hz, 1 H), 7.1 1 (dd, J = 8.5, 0.4 Hz, 1 H), 4.36 (q, J = 7.2 Hz, 2H), 2.54 (s, 3H), 1 .40 (t, J = 7.0 Hz, 3H).
Example E3-1 -2 / PKE 490: 5-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester: Method as for Bl -1 -1 , using 2,2'-bithiophene-5-boronic acid and 5-bromo-2-methyl-benzoic acid ethyl ester E3-1 -1 ; white solid (36%). 1 H NMR (CDCI3) δ 8.1 2 (d, J = 2.1 Hz, I H), 7.60 (dd, J = 7.9, 2.1 Hz, I H), 7.27- 7.20 (m, 4H), 7.1 5 (d, J = 3.8 Hz, 1 H), 7.03 (dd, J = 5.1 , 3.6 Hz, 1 H), 4.40 (q, J = 7.1 Hz, 2H), 2.60 (s, 3H), 1 .43 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 329.1 [M + H]+. Mp: 55°C-57 °C.
Example E3-2 / PKE 531 : 5-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid: Method as for B2-1 , using 5- [2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester E3-1 -2; pale yellow solid (86%). 1 H NMR (DMSO) δ 8.03 (d, J = 2.3 Hz, 1 H), 7.75 (dd, J = 7.9, 2.1 Hz, 1 H), 7.53 (dd, J = 5.2, 1 .1 Hz, 1 H), 7.51 (d, J = 3.9 Hz, 1 H), 7.37 (m, 2H), 7.32 (d, J = 3.7 Hz, 1 H), 7.1 1 (dd, J = 5.1 , 3.6 Hz, 1 H), 2.53 (s, 3H). MS (ESI+) m/z: 301 .1 [M + H]+. Mp: 231 °C-234°C.
Example E3-3-1 : 2-Methyl-5-thiophen-2-yl-benzoic acid ethyl ester: Method as for Bl -1 -1 , using thio- phene-2-boronic acid and 5-bromo-2-methyl-benzoic acid ethyl ester E3-1 -1 ; orange oil (78%). 1 H NMR (CDCI3) δ 8.14 (d, J = 2.2 Hz, I H), 7.62 (dd, J = 7.9, 2.1 Hz, 1 H), 7.33 (dd, J = 3.5, 1 .1 Hz, I H), 7.29 (dd, J = 5.1 , 1 .1 Hz, 1 H), 7.25 (d, J = 7.0 Hz, 1 H), 7.08 (dd, J = 5.1 , 3.6 Hz, 1 H), 4.39 (q, J = 7.1 Hz, 2H), 2.60 (s, 3H), 1 .42 (t, J = 7.1 Hz, 3H).
Example E3-3-2: 5-(5-Bromo-thiophen-2-yl)-2-methyl-benzoic acid ethyl ester: Method as for Bl -1 -2, using 2-methyl-5-thiophen-2-yl-benzoic acid ethyl ester E3-3-1 ; orange oil (98%). 1 H NMR (CDCI3) δ 8.04 (d, J = 2.1 Hz, 1 H), 7.51 (dd, J = 8.0, 2.1 Hz, 1 H), 7.24 (d, J = 8.3 Hz, 1 H), 7.06 (d, J = 3.8 Hz, 1 H), 7.03 (dd, J = 3.8, 1 .1 Hz, 1 H), 4.39 (q, J = 7.1 Hz, 2H), 2.59 (s, 3H), 1 .41 (t, J = 7.1 Hz, 3H). Example E3-3-3 / PKE 532: 5-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-2-methyl-benzoic acid ethyl ester
Figure imgf000078_0001
Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 5-(5-Bromo-thiophen-2-yl)-2-methyl- benzoic acid ethyl ester E3-3-2; yellow solid (36%). 1 H NMR (CDCI3) δ 8.1 5 (d, J = 1 .9 Hz, I H), 7.80- 7.72 (m, 2H), 7.63 (dd, J = 8.1 , 2.1 Hz, 1 H), 7.43 (s, 1 H), 7.35-7.27 (m, 5H), 4.41 (q, J = 7.0 Hz, 2H), 2.62 (s, 3H), 1 .45 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 379.2 [M + H]+. Mp: 1 52 °C-1 55°C.
Example E3-4 / PKE 539: 5-(5-Benzo[b]thiophen-2-yl-thiophen-2-yl)-2-methyl-benzoic acid: Method as for B2-1 , using 5-(5-benzo[b]thiophen-2-yl-thiophen-2-yl)-2-methyl-benzoic acid ethyl ester E3-3-3; white solid (93%). λ H NMR (DMSO) δ 7.99 (d, J = 2.2 Hz, 1 H), 7.96 (dd, J = 9.0, 1 .5 Hz, 1 H), 7.83 (dd, J = 6.3, 1 .4 Hz, 1 H), 7.71 (m, 1 H), 7.69 (s, 1 H), 7.56 (d, J = 3.8 Hz, 1 H), 7.47 (d, J = 4.0 Hz, 1 H), 7.38 (ddd, J = 7.0, 5.5, 1 .7 Hz, 2H), 7.31 (d, J = 7.8 Hz, 1 H). MS (ESI+) m/z: 351 .1 [M + H]+. Mp: 278°C- 283°C.
Example E3-5 / PKE 503: 4-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester: Method as for Bl - 1 -1 , using 2,2'-bithiophene-5-boronic acid and 4-bromo-2-methyl-benzoic acid ethyl ester; yellowish solid (68%). λ H NMR (CDCI3) δ 7.95 (dd, J = 8.6, 1 .5 Hz, 1 H), 7.48-7.45 (m, 2H), 7.32 (dd, J = 3.8, 1 .8 Hz, I H), 7.26-7.21 (m, 2H), 7.1 7 (dd, J = 3.8, 1 .7 Hz, I H), 7.04 (ddd, J = 5.1 , 3.6, 1 .7 Hz, I H), 4.36 (q, J = 7.1 Hz, 2H), 2.65 (s, 3H), 1 .41 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 329.1 [M + H]+. Mp: 80°C-81 0C.
Example E3-6 / PKE 502: 4-[2,2 ']Bithiophenyl-5-yl-2-methyl-benzoic acid: Method as for B2-1 , using 4- [2,2 ']bithiophenyl-5-yl-2-methyl-benzoic acid ethyl ester E3-5; yellow solid (83%). 1 H NMR (DMSO) δ 7.55-7.50 (m, 2H), 7.44 (d, J = 3.7 Hz, 1 H), 7.37-7.32 (m, 3H), 7.29 (d, J = 3.8 Hz, 1 H), 7.1 0 (dd, J = 5.1 , 3.7 Hz, I H), 2.47 (s, 3H). MS (ESI+) m/z: 301 .1 [M + H]+. Mp: 305°C-308°C.
Example E4-1 -1 : 5-Bromo-4-methyl-1 ,3-thiazole-2-carboxylic acid ethyl ester: To a solution of ethyl-4- methyl-1 ,3-thiazole-2-carboxylate (300 mg, 1 .7 mmol) in acetonitrile (1 0 ml) at room temperature is added portion wise /V-bromosuccinimide (3 times 31 0 mg over 24 hours, 6.0 mmol). Reaction mixture is stirred at room temperature for 48 h. Solvents were evaporated and the residue was purified by silica gel column chromatography (EtOAc/cyclohexane: 2/8) to afford a yellow oil (65%). 1 H NMR (CDCI3) δ 4.46 (q, J = 7.2 Hz, 2H), 2.49 (s, 3H), 1 .42 (t, J = 7.1 Hz, 3H).
Example E4-1 -2: 4-Methyl-5-thiophen-2-yl-1 ,3-thiazole-2-carboxylic acid ethyl ester: Method as for Bl -I - 1 , using thiophene-2-boronic acid and 5-bromo4-methyl-thiazole-2-carboxylic acid ethyl ester E4-1 -1 ; yellow solid (46%). λ H NMR (CDCI3) δ 7.43 (dd, J = 5.1 , 1.1 Hz, 1 H), 7.25 (dd, J = 3.8, 1.3 Hz, 1 H), 7.12 (dd, J = 5.1 , 3.6 Hz, 1 H), 4.49 (q, J = 7.2 Hz, 2H), 2.67 (s, 3H), 1.44 (t, J = 7.2 Hz, 3H).
Example E4-1-3: 5-(5-Bromo-thiophen-2-yl)4-methyl-l,3-thiazole-2-carboxylic acid ethyl ester: Method as for Bl-I -2, using 4-methyl-5-thiophen-2-yl-thiazole-2-carboxylic acid ethyl ester E4-1-2; yellow solid (96%). λ H NMR (CDCI3) δ 7.08 (d, J = 3.8 Hz, 1 H), 6.99 (d, J = 3.8 Hz, 1 H), 4.48 (q, J = 7.1 Hz, 2H), 2.63 (s, 3H), 1.44 (t, J = 7.1 Hz, 3H).
Example E4-1-4/ PKE 538: 5-(5-Benzo[b]thiophen-2yl-thiophen-2-yl)-4-methyl-thiazole-2-carboxylic acid ethyl ester: Method as for Bl -2, using benzo[b]thiophene-2-boronic acid and 5-(5-Bromo-thiophen-2-yl)-4- methyl-thiazole-2-carboxylic acid ethyl ester E4-1-3; yellow solid (94%).1H NMR (CDCI3) δ 7.81-7.74 (m, 2H), 7.46 (d, J = 0.6 Hz, 1 H), 7.37-7.33 (m, 2H), 7.29 (d, J = 4.0 Hz, 1 H), 7.20 (d, J = 3.8 Hz, 1 H), 4.49 (q, J = 7.1 Hz, 2H), 2.73 (s, 3H), 1.45 (t, J = 7.1 Hz, 3H). MS (ESI+) m/z: 386.1 [M + H]+. Mp: 185°C-188°C.
The above-described embodiments of the present invention are intended to be examples only. Alterations, modifications and variations may be effected to the particular embodiments by those of skill in the art without departing from the scope of the invention, which is defined solely by the claims appended hereto. All references are herein incorporated by reference.
REFERENCES
1. https://www.who.int/csr/disease/influenza/vaccinerecommendationsl/en/. 2. Moscona, A. N Engl J Med 353, 1363-1373 (2005).3. Davies, Wl. et al. Science 144, 862-863 (1964).4. Moscona, A. N Engl J Med 360, 953-956 (2009).5. Dharan, NJ. et al. JAMA 301, 1034-1041 (2009).6. Pilger, B.D., Cui, C. & Coen, D. M. Chem Biol 11, 647-654 (2004).7. Brownlee, CC. & Sharps, JI. J Virol 76, 7103-7113 (2002).8. Perales, B. & Ortin, J. J Virol 71, 1381-1385 (1997).9. Fodor, E. etal. J Virol 76, 8989-9001 (2002). 10. He, X. et al. Nature 454, 1123-1126 (2008). 11. Obayashi, E. et al. Nature 454, 1127-1131 (2008).12. Chanem, A. et al. J Virol 81 , 7801-7804 (2007). 13. Dostmann, W.R. et al. Proc Natl Acad Sci U S A 97, 14772-14777 (2000).14. Chokephaibulkit, K. et al. Pediatr Infect Dis J 24, 162-166 (2005).15. Norton, CP. et al. Virology 156, 204-213 (1987).16. Schwemmle, M. et al., P. Virology 206, 545-554 (1995).17. Mayer, D. etal. J Proteome Res 6, 672-682 (2007).18. Pleschka, S. etal. J Virol 70, 41884192 (1996). 19. Schneider, U. etal., M. J Virol 77, 11781-11789 (2003).20. Schmidtke, M., etal., J Virol Methods 95, 133-143 (2001).21. Edgar, R.C. Nucleic Acids Res 32, 1792- 1797 (2004).

Claims

Claims
1. A compound represented by one of formulas (Xl) to (XXIII)
Figure imgf000080_0001
(XIX) (XX)
Figure imgf000081_0001
where X is sulfur or oxygen; where R2 is selected from a group consisting of: hydrogen, cyano, formyl, acetyl, ethylcarbonyl, pro- pylcarbonyl, isopropylcarbonyl, carboxy, carboxy methyl, methoxycarbonyl, ethoxycarbonyl, propoxy- carbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, methoxycar- bonylmethyl, methoxycarbonylethyl, ethoxycarbonylmethyl, methoxycarbonyl ethyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trihalogenomethyl, difluoromethyl, diha- logenomethyl, fluoromethyl, halogenomethyl, phenyl, 1 -naphthyl, 2-naphthyl, aryl, hydroxy methyl, 1 -hydroxyethyl, 2-hydroxyethyl, carbamoyl, hydroxycarbamoyl, methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, isopropylcarbamoyl, butylcarbamoyl, isobu-tylcarbamoyl, tert-butylcarbamoyl, (1- (2-thienylcarbonyl)-pyrazol-5-yl), (l -(m-trifluoromethyl-benzoyl)-pyrazol-5-yl), (l -(tert-butylcarbonyl)- pyrazol-5-yl), (l -ethyl-tetrahydropyrrol-2-yl)-methyl-carbamoyl, (l -methyl-imidazol4-yl)- sulfonamidomethyl, (2-morpholinoethoxy)-carbonyl, (2-morpholinoethyl)-carbamoyl, (3-(imidazol-l - yl)-propyl)-carbamoyl, (3-morpholinopropyl)-carbamoyl, (4-(p-fluorophenyl)-2,3,5,6-tetrahydropyrazin- l -yl)-carbonyl, (tetrahydropyrrol-l -yl)-carbonyl, 2-benzofuranyl, benzamidomethyl, cyclopropylami- domethyl, C1 -C4 alkylami-domethyl, benzamidoethyl, cyclopropylamidoethyl, C1 -C4 alkylami- doethyl, phenylsul-fonamidomethyl, cyclosulfonamidomethyl, C1 -C4 alkylsulfonamidomethyll, phenylsul-fonamidoethyl, cyclosulfonamidoethyl, and C1 -C4 alkylsulfonamidoethyl, 1 W-tetrazol-5- yl, 2W-tetrazol-5-yl, l W-l -Z1 0-tetrazol-5-yl, 2H-] -Zl 0-tetrazol-5-yl, where Zl O is selected from a group consisting of: methyl, C2-C4 alkyl, and substituted alkyl chains; where R3 is selected from a group consisting of: hydrogen, formyl, acetyl, ethylcarbonyl, propylcar- bonyl, isopropylcarbonyl, carboxy, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxy-carbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trihalogenomethyl, difluromethyl, dihalogenomethyl, fluoromethyl, halogenomethyl, hydrazinocarbonyl, m,p-difluorophenyl, m,p-dihalogenophenyl, p- (trifluoromethyl)-phenyl, p-(tιϊhalogenomethyl)-phenyl, p-chlorophenyl, p-halogenophenyl, and p- nitrophenyl; where R4 is selected from a group consisting of: methyl, C2-C4 alkyl; where Q2 is selected from a group consisting of: hydrogen, 2-thienyl, 5-acetyl-thien-2-yl, and aryl; where Q3 is selected from a group consisting of: hydrogen, phenyl, 1 -naphthyl, 2-naphthyl, methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, tert-butyl, bromo, halogeno, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2- thienyl, 3-thienyl, 2-furyl, 3-furyl, 4-pyrazolyl, benzo[b]furan-2-yl, benzo[b]thiophen-2yl, 3-quinolyl, 1 - isoquinolyl, 4-isoquinolyl, 2-indolyl, 5-pyrimidinyl, (2-thienyl)-carbonyl, l-(tert-butoxycarbonyl)-indol- 2-yl, 1 ,2-oxazol-5-yl, l ,3-benzodioxol-5-yl, l ,3-thiazol-2-yl, l -methylpyrazol4-yl, 2,4-dimethyl-l ,3- thiazol-5-yl, 4-methyl-2-phenyl-l ,3-thiazol-5-yl, 4-phenyl-thien-2-yl, 5-(2-thienyl)-thien-2-yl, 3-methyl- thien-2-yl, 3-ethyl-thien-2-yl, 4-methyl-thien-2-yl, 4-ethyl-thien-2-yl, o-Q7-phenyl, m-Q7-phenyl, p-Q7- phenyl, o,m-bis(Q7)-phenyl, o,p-bis(Q7)-phenyl, m,p-bis(Q7)-phenyl, 5-Q7-benzo[b]thiophen-2-yl, 5-Q7- benzo[b]furan-2-yl, 5-Q7-benzo[b]thiophen-3-yl, 5-Q7-benzo[b]furan-3-yl, 5-Q7-benzo[b]thiophen-5-yl, 5-Q7-benzo[b]furan-5-yl, 6-Q7-benzo[b]thiophen-3-yl, 6-Q7-benzo[b]furan-3-yl, 6-Q7-benzo[b]thiophen- 5-yl, 6-Q7-benzo[b]furan-5-yl, 6-Q7-naphth-2-yl, and 5-Q7-thien-2-yl, wherein Q7 is selected from a group consisting of: hydroxy, trifluoromethoxy, trifluoromethoxy, trihalogenomethoxy, difluoro- methoxy, dihalogenomethoxy, fluoromethoxy, halogenomethoxy, trifluoromethyl, trihaloge- nomethyl, difluoromethyl, dihalogenomethyl, fluoromethyl, halogenomethyl, formyl, acetyl, chloro, fluoro, halogeno, cyano, nitro, methyl, ethyl, C3-C4 alkyl, methoxy, ethoxy, and C3-C4 alkoxy; where Q4, Q5, and Z9 are independently selected from a group consisting of: hydrogen, methyl, ethyl, C3-C4 alkyl, and phenyl; where Z2 is selected from a group consisting of: hydrogen, methyl, ethyl, C3-C4 alkyl, methoxy, ethoxy, and C3-C4 alkoxy; where Z5, Z6, and Z7 are independently selected from a group consisting of: hydrogen, phenyl, p- tolyl, 1 -naphthyl, 2-naphthyl, 2-thienyl, 2-furyl, benzo[b]thiophen-2-yl, benzo[b]furan-2-yl, and hydroxy; and where Z8 is selected from a group consisting of: bromo. halogeno, benzo[b]thiophen-2-yl, and benzo[b]furan-2-yl; or a physiologically tolerable salt, solvate, or physiologically functional derivative thereof; wherein said compound is not one of the following:
1 -{4-Methyl-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazol-5-yl}ethan-l -one; 4-(4-nitrophenyl)-2-[5-(2-thienyl)-2- thienyl]-l ,3-thiazole; 4-(4-chlorophenyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole; 2-[5-(2-thienyl)-2- thienyl]-4-[4-(trifluoromethyl)phenyl)-l ,3-thiazole; 4-(3,4-difluorophenyl)-2-[5-(2-thienyl)-2-thienyl]- 1 ,3-thiazole; 4-(tert-butyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole; (2-thienyl)(5-{2-[5-(2-thienyl)-2- thienyl]-4-methyl-l ,3-thiazol-5-yl}-l H-pyrazol-1 -yl)methanone; 1 -(5-{2-[5-(2-thienyl)-2-thienyl]-4- methyl-l ,3-thiazol-5-yl}-l H-pyrazol-1 -yl)-2,2-dimethylpropan-l -one; (5-{2-[5-(2-thienyl)-2-thienyl]-4- methyl-1 ,3-thiazol-5-yl}-l H-pyrazol-1 -yl)[3-(trifluoromethyl)phenyl]methanone; 5-[5-(4-benzo[b]furan- 2-yl-l ,3-thiazol-2-yl)thien-2-yl]isoxazole; methyl [2-(5-isoxazol-5-ylthien-2-yl)-5-methyl-l ,3-thiazol-4- yl]acetate; 2-[5-(4-phenyl-l ,3-thiazol-2-yl)-2-thienyl]pyridine; 2-[5-[4-(tert-butyl)-l ,3-thiazol-2-yl]-2- thienyl}pyridine; ethyl 2-(5-isoxazol-5-ylthien-2-yl)-4-methyl-l ,3-thiazole-5-carboxylate; 4-methyl-2-[4- phenyl-5-(trifluoromethyl)-2-thienyl]-l ,3-thiazole-5-carboxylic acid; N-{[2-(2-thienyl)-l ,3-thiazol-4- yljmethyl} benzenesulfonamide; N-{[2-(2-thienyl)-l ,3-thiazol4-yl]methyl} benzamide; l -[4-methyl-2-
(2-thienyl)-l ,3-thiazol-5-yl]ethan-l -one; 4-methyl-5-(l H-pyrazol-5-yl)-2-(2-thienyl)-l ,3-thiazole; 4- methyl-5-[2-(methylthio)pyrimidin4-yl]-2-(2-thienyl)-l ,3-thiazole; cyclopropyl-{5-[4-methyl-2-(2- thienyl)-l ,3-thiazol-5-yl]-l H-pyrazol-l -yl}methanone; N l -isopropyl-5-[4-methyl-2-(2-thienyl)-l ,3- thiazol-5-yl]-l H-pyrazole-1 -carboxamide; 1 -methyl-N-{[2-(2-thienyl)-l ,3-thiazol4-yl]methyl}-l H- imidazole-4-sulfonamide; N-{[2-(2-thienyl)-l ,3-thiazol4-yl] methyl} cyclopropanecarboxamide; 4- methyl-2-(2-thienyl)-l ,3-thiazole-5-carbohydrazide.
2. The compound according to claim 1 as a medicament.
3. A compound as a medicament, wherein the compound is one of the following: 1 -{4-Methyl-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazol-5-yl}ethan-l -one,
4-(4-nitrophenyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole,
4-(4-chlorophenyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole,
2-[5-(2-thienyl)-2-thienyl]4-[4-(trifluoromethyl)phenyl)-l ,3-thiazole,
4-(3,4-difluorophenyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole, 4-(tert-butyl)-2-[5-(2-thienyl)-2-thienyl]-l ,3-thiazole,
(2-thienyl)(5-{2-[5-(2-thienyl)-2-thienyl]4-methyl-l ,3-thiazol-5-yl}-l H-pyrazol-l -yl)methanone, l -(5-{2-[5-(2-thienyl)-2-thienyl]4-methyl-l ,3-thiazol-5-yl}-l H-pyrazol-l -yl)-2,2-dimethylpropan-l -one,
(5-{2-[5-(2-thienyl)-2-thienyl]4-methyl-l ,3-thiazol-5-yl}-l H-pyrazol-1 -yl)[3-
(trifluoromethyl)phenyl]methanone, 5-[5-(4-benzo[b]furan-2-yl-l ,3-thiazol-2-yl)thien-2-yl]isoxazole, methyl [2-(5-isoxazol-5-ylthien-2-yl)-5-methyl-l ,3-thiazol4-yl]acetate,
2-[5-(4-phenyl-l ,3-thiazol-2-yl)-2-thienyl] pyridine,
2-{5-[4-(tert-butyl)-l ,3-thiazol-2-yl]-2-thienyl}pyridine, ethyl 2-(5-isoxazol-5-ylthien-2-yl)4-methyl-l ,3-thiazole-5-carboxylate, 4-methyl-2-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-l ,3-thiazole-5-carboxylic acid,
N-{[2-(2-thienyl)-l ,3-thiazol4-yl]methyl} benzenesulfonamide,
N-{[2-(2-thienyl)-l ,3-thiazol4-yl]methyl} benzamide, l -[4-methyl-2-(2-thienyl)-l ,3-thiazol-5-yl]ethan-l -one,
4-methyl-5-(l H-pyrazol-5-yl)-2-(2-thienyl)-l ,3-thiazole, 4-methyl-5-[2-(methylthio)pyrimidin-4-yl]-2-(2-thienyl)-l ,3-thiazole, cyclopropyl{5-[4-methyl-2-(2-thienyl)-l ,3-thiazol-5-yl]-l H-pyrazol-1 -yl}methanone, N l -isopropyl-5-[4-methyl-2-(2-thienyl)-l ,3-thiazol-5-yl]-l H-pyrazole-1 -carboxamide, 1 -methyl-N-{[2-(2-thienyl)-l ,3-thiazol4-yl]methyl}-l H-imidazole-4-sulfonamide, N-{[2-(2-thienyl)-l ,3-thiazol4-yl] methyl} cyclopropanecarboxamide, 4-methyl-2-(2-thienyl)-l ,3-thiazole-5-carbohydrazide; or a physiologically tolerable salt, solvate, or physiologically functional derivative thereof.
4. The compound according to any of claims 1 to 3 for treating and/or preventing influenza type A and/or influenza type B infections in humans, mammals and/or birds.
5. The compound according to any of claims 1 to 3 for treating and/or preventing respiratory syncytial virus infections in humans, mammals and/or birds.
6. A use of the compound according to any of claims 1 to 3 for the manufacture of a medicament for the treatment and/or prevention of influenza type A and/or influenza type B infections in humans, mammals and/or birds.
7. A use of the compound according to any of claims 1 to 3 for the manufacture of a medicament for the treatment and/or prevention of respiratory syncytial virus infections in humans, mammals and/or birds.
8. A pharmaceutical composition comprising a compound according any of claims 1 to 5.
9. A pharmaceutical composition according to claim 8, with one or more excipients.
1 0. A method for identifying small molecules and/or peptidic inhibitors or disruptors for PPIs comprising the following steps a) selection of a proteome of interest b) identification of potential target regions of PPI inhibitors c) detection of drugable protein-binding domains d) synthesis of peptides corresponding to the druggable protein-binding domains e) testing of the synthesized peptides for their ability to bind the protein partner involved in the PPI f) development of a binding assay for the identification of small molecules and/or peptidic inhibitors or disruptors based on the positively tested peptides g) high-throughput screening with the binding assay and selecting the active small molecules and/or peptidic inhibitors or disruptors.
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