JP2006070014A - 2-aminobenzothiazole derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new benzothiazole derivative having excellent antibacterial activity to funguses, fungistatic activity excellent in safety, and N-myristoyl transferase inhibiting activity. <P>SOLUTION: The medicines contain a 2-aminobenzothiazole derivative represented by formula (I) (wherein, R<SP>1</SP>, R<SP>2</SP>and X are a substituent) or a pharmaceutically permissible salt thereof as an active ingredient. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a novel 2-aminobenzothiazol derivative having antifungal activity and N-myristoyltransferase inhibitory activity.

  For mycosis, superficial mycosis represented by various ringworms, scabies, psoriasis, cutaneous candidiasis, mycotic meningitis, fungal respiratory infection, fungiemia, urinary tract mycosis, etc. There is a representative mycosis. Among these, deep mycosis such as candidiasis and aspergillosis has been increasing in recent years due to the frequent use of cancer chemotherapeutic agents and immunosuppressive agents, and decreased in vivo immunity due to HIV infection, etc. Drugs effective against bacteria are desired. Conventionally, amphotericin B and azole compounds fluconazole, itraconazole, etc. are known as effective drugs against Aspergillus and Candida, but they are sufficiently satisfactory in terms of antifungal spectrum, pharmacokinetics, resistance, etc. is not. In addition, azole drugs that have been studied for many years in this area are problematic in terms of their effectiveness in serious patients and the resistance to long-term use of the same drugs. On the other hand, development of a compound having excellent antibacterial activity against deep mycosis and excellent safety is desired.

N-myristoyltransferase is an enzyme that transfers a myristoyl group from myristoyl CoA to the N-terminal glycine of eukaryotic and virus-derived proteins. Many of N-myristoylated proteins are physiologically active proteins involved in cell signal transduction, and by being myristoylated, they are localized on the cell membrane and functional expression is possible. From genetic studies, it is known that this enzyme is an essential enzyme for the survival of pathogenic fungi, for example, an essential enzyme for the survival of Candida albicans (Non-patent Document 1). For this reason, N-myristoyltransferase inhibitors are considered as target proteins in the development of fungicidal antifungal agents. In fact, it has been reported that a benzofuran derivative having an inhibitory action on this enzyme has an antifungal action (Non-Patent Documents 2, 3 and the like). On the other hand, it has been reported that these compounds have problems in gastrointestinal stability and pharmacokinetics. Therefore, development of a more stable and effective N-myristoyltransferase inhibitor is desired.
Weinberg et al .; Molecular Microbiology 1995, 16, 241-250. Devadas et al .; J. Med. Chem. 1997, 40, 2609-2625. Ebiike et al .; Bioorganic & Medicinal Chemistry Letters 2002, 12, 607-610

  An object of the present invention is to provide a compound having an excellent antibacterial activity against fungi, an excellent N-myristoyltransferase inhibitory activity, and useful as an antifungal agent.

  Accordingly, the present inventors have synthesized a number of benzothiazol compounds and studied their pharmacological actions. As a result, the present inventors have demonstrated excellent N-myristoyltransferase inhibitory activity and compounds having a structure represented by the following formula (1). It has been found that it has antifungal activity and has completed the present invention.

  That is, the present invention provides the following general formula (I)

(Wherein R ¹ represents a cycloalkyl group, an aromatic hydrocarbon group which may have a substituent, or an aromatic heterocyclic group containing a nitrogen atom, an oxygen atom or a sulfur atom which may have a substituent). R ² has a lower alkyl group, an aralkyl group which may have a substituent, a cycloalkyl group, an oxiranyl group, an aromatic hydrocarbon group which may have a substituent or a substituent. Or an aromatic heterocyclic group containing a nitrogen atom, oxygen atom or sulfur atom, wherein X represents a group —Y—CH (R ³ ) —Z—N (R ⁴ ) —CO— or a group —Z—CONH—. Y represents a group — (CH ₂ ) _n — (where n represents 0 or 1), Z represents a linear or branched lower alkylene group, or a 5- to 6-membered cycloalkyl group. , R ³ and R ⁴ each independently represent a hydrogen atom, or a lower alkyl group, or R ³ and R ⁴ Shows the saturated heterocyclic group of 4 to 6-membered ring containing a nitrogen atom together)
The 2-aminobenzothiazole derivative represented by these, or its pharmaceutically acceptable salt is provided.
Moreover, this invention provides the pharmaceutical which uses the 2-aminobenzothiazole derivative or its pharmaceutically acceptable salt represented with general formula (I) as an active ingredient.

  The 2-aminobenzothiazole derivative represented by the general formula (I) of the present invention has an excellent antibacterial activity against fungi and an excellent N-myristoyltransferase inhibitory activity. Useful as a fungicide.

  The cycloalkyl group in the general formula (I) is preferably a cycloalkyl group having 3 to 9 carbon atoms, and more preferably 3 to 7 carbon atoms. Specific examples include cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group and the like.

  As the lower alkyl group, a linear or branched alkyl group having 1 to 6 carbon atoms and more preferably 1 to 5 carbon atoms is preferable. Specific examples include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group and the like.

The aralkyl group is preferably a group in which an alkyl group having 1 to 5 carbon atoms is bonded to an aryl group having 6 to 14 carbon atoms, such as a phenyl-C _1-5 -alkyl group or a naphthyl-C _1-5 -alkyl group. Can be mentioned. More specifically, a benzyl group, a phenethyl group, etc. are mentioned. In the alkyl group, an alkyl group having 1 to 6 carbon atoms, a trifluoromethyl group, a halogen atom, or the like may be substituted by 1 to 4 carbon atoms.

  Examples of the aromatic hydrocarbon group include an aryl group having 6 to 14 carbon atoms. Specific examples include a phenyl group, a naphthyl group, an indenyl group, and a fluorenyl group. The aromatic heterocyclic group is a 5- or 6-membered aromatic heterocyclic ring containing a nitrogen atom, an oxygen atom or a sulfur atom. Specifically, a pyrrolyl group, a pyrazolyl group, an imidazolyl group, a furanyl group, Examples include oxazolyl group, thienyl group, thiazolyl group, pyridyl group, pyrimidinyl group and the like. These aromatic hydrocarbon groups and aromatic heterocyclic groups may have a substituent. Examples of the substituent include an alkyl group having 1 to 6 carbon atoms which may be substituted by halogen, and 1 carbon atom. -6 alkoxy group, halogen atom, optionally substituted cycloalkyl group, hydroxyl group, nitro group, cyano group, alkylthio group having 1 to 6 carbon atoms, halogen optionally substituted 6 to 6 carbon atoms 14 aryl groups and the like, and 1 to 4 of them may be substituted. Here, a cycloalkyl group, an aryl group, etc. are the same as the above.

  Examples of the linear or branched lower alkylene group represented by Z include an alkylene group having 1 to 6 carbon atoms, such as a methylene group, an ethylene group, a trimethylene group, and a tetramethylene group. Examples of the 5- to 6-membered cycloalkyl group include a cyclopentyl group and a cyclohexyl group.

Examples of the saturated heterocycle formed by —CH (R ³ ) —ZN (R ⁴ ) — include azetidine, pyrrolidine, and piperidine.

  The 2-aminobenzothiazole derivative represented by the general formula (I) of the present invention includes its racemate, optical isomer and isotope.

  Examples of the salt of the 2-aminobenzothiazole derivative of the present invention include hydrochloride, nitrate, hydrobromide, p-toluenesulfonate, methanesulfonate, fumarate, maleate, malonate, and succinate. Acid salts, citrate salts, tartrate salts and the like.

  The 2-aminobenzothiazol derivative represented by the general formula (I) of the present invention can be produced, for example, by the method described in Reaction Formulas A to C shown below. Here, those skilled in the art can easily select the solvent, reaction temperature, pressure, and other reaction conditions, and are not particularly limited. As the starting materials, commercially available compounds are appropriately used, and they are produced by a known production method.

  Reaction formula A is an example of a general production method of a 2-aminobenzothiazole derivative represented by general formula (Ia).

2-aminobenzothiazole-6-carboxylic acid ester (2) can be prepared by known methods (Stuckwisch, C.g .; J. Am. Chem. Soc. 1949, 71, 3417, Ismail, i.A .; Sharp, d.E .; Chedekel, MR; see J. Org. Chem. 1980, 45, 2243).
The obtained compound (2) is a carboxylic acid (R ² COOH) or its reactivity under known conditions for forming an amide bond (see, for example, Experimental Chemistry Course 22, Organic Synthesis IV Acid, Amino Acid, Peptide, 4th Edition). Compound (3) is obtained by conducting a condensation reaction with a derivative (for example, carboxylic acid anhydride ((R ² CO) ₂ O) or carboxylic acid chloride (R ² COCl)).
Carboxylic acid (4) is obtained by carrying out alkali hydrolysis of the ester part of compound (3). As the base for alkali hydrolysis, sodium hydroxide, potassium hydroxide, lithium hydroxide aqueous solution or the like can be used. The solvent is preferably an alcohol solvent such as methanol or ethanol.
The carboxamide derivative (5) is obtained by condensing the obtained carboxylic acid (4) with a primary amine or a secondary amine in the same manner as the synthesis of the compound (3) described above.
The carboxamide derivative (5) was obtained by removing the t-butoxycarbonyl group (described as Boc in the reaction formula) using an acid catalyst such as hydrochloric acid or trifluoroacetic acid in a solvent such as dioxane or dichloromethane to obtain a compound (6) To obtain a salt of hydrochloric acid, trifluoroacetic acid, etc.

Reductive amination reaction of compound (6) or a salt thereof with aldehyde (R ¹ CHO), that is, sodium triacetoxyborohydride (Maryanoff et al., J. Org. Chem. 1996) in a solvent such as methanol, dichloromethane, dichloroethane, dimethylformamide and the like. 61, 3849), sodium cyanoborohydride, triacetoxyborohydride supported on polystyrene (see Bhattacharya et al., Tetrahedron Lett. 2003, 44, 4957) and the like in the presence of a reducing agent (I) The 2-aminobenzothiazole derivative shown by is obtained. In particular, the reaction using triacetoxyborohydride supported on polystyrene as a reducing agent has an advantage that the target product can be easily isolated and purified, and parallel synthesis using a synthesis robot is possible.

  Further, the 2-aminobenzothiazole derivative represented by the general formula (Ia) can also be produced by using the reaction formula B shown below.

Reduction of compound (7) and aldehyde (R ¹ CHO), such as sodium cyanoborohydride, sodium triacetoxyborohydride, triacetoxyborohydride supported on polystyrene, in a solvent such as methanol, dichloromethane, dichloroethane, dimethylformamide, etc. Compound (8) is obtained by carrying out the reaction in the presence of an agent, and then compound (9) is obtained by protecting the secondary amine moiety with an allyl carbamate group (described as Alloc in the reaction formula).
Compound (9) is obtained by removing the t-butoxycarbonyl group (described as Boc in the reaction formula) using an acid catalyst such as hydrochloric acid or trifluoroacetic acid in a solvent such as dioxane or dichloromethane to obtain compound (10). .
Compound (10) is condensed with a benzothiazole-6-carboxylic acid derivative in the presence of a condensing agent in a solvent such as dimethylformamide or dimethylacetamide to obtain carboxamide derivative (11).
Further, the t-butoxycarbonyl group is removed by the above method to obtain the compound (12), and then condensed with various carboxylic acids (R ¹ COOH) or reactive derivatives thereof to obtain the compound (13).

  Finally, by deprotecting the allyl carbamate group in the presence of a suitable carbon nucleophile (for example, dimedone, barbituric acid, solid-supported barbituric acid (see Tsukamoto et al .; Synlett 2003, 1105)) palladium catalyst, A 2-aminobenzothiazole derivative represented by the general formula (I) is obtained. In particular, in the deprotection reaction using solid-supported barbituric acid such as Tsukamoto et al., It is very easy to isolate and produce the final product, and a parallel reaction using a synthetic robot is also possible.

  The following reaction formula C is an example of a general production method of a 2-aminobenzothiazole derivative represented by the general formula (Ib).

Compound (16) can be produced by carrying out the reaction in the same manner as in reaction formula (A) except that compound (14) is used as the starting material.
Compound (16) is subjected to a catalytic hydrogenation reaction using a transition metal catalyst such as palladium carbon to obtain amine (17). For the reaction, an alcohol solvent such as methanol or ethanol is suitable. The obtained amine (17) is condensed with an amino acid derivative in the presence of a condensing agent in a solvent such as dimethylformamide or dimethylacetamide to obtain a compound (18). Thereafter, the 2-aminobenzothiazole derivative represented by the general formula (Ib) can be produced by a method similar to the reaction formula A.

  Isolation of the 2-aminobenzothiazol derivative of the present invention from the reaction mixture can be carried out by means such as liquid-liquid extraction, solid-phase extraction, recrystallization, column chromatography, ion exchange and the like.

The 2-aminobenzothiazole derivative of the present invention and a pharmaceutically acceptable salt thereof are excellent in N-myristoyltransferase inhibitory activity and antifungal activity, and generally contain carriers, excipients and additives used for pharmaceutical formulation. It is prepared into tablets, powders, fine granules, fruit granules, capsules, pills, solutions, injections, suppositories, vaginal preparations, ointments, patches and the like, and is orally or parenterally administered.
In order to produce a solid preparation, the 2-aminobenzothiazol of the present invention and a pharmaceutically acceptable salt thereof are mixed with an excipient, and if necessary, a binder, a disintegrant, a bulking agent, a coating agent, It is preferable to add tablets, granules, capsules, suppositories, etc. by conventional methods after adding a sugar coating or the like.
When preparing an injection, the 2-aminobenzothiazol of the present invention and a pharmaceutically acceptable salt thereof are prepared as a solution prepared by dissolving, dispersing or emulsifying in an aqueous carrier such as distilled water for injection in advance. It is preferable to use an injectable powder for dissolution at the time of use. Examples of the administration method of the injection include intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intravenous infusion and the like.

  The dose of the 2-aminobenzothiazol derivative of the present invention to humans varies depending on the state of infection and the administration method. For example, when administered to an adult patient for the purpose of treating Candida infection, The 2-aminobenzothiazole derivative of the present invention is about 0.01-100 mg / kg / day, preferably about 0.1-50 mg / kg / day, more preferably about 1-20 mg / kg / day. is there.

  Reference Examples and Examples will be described below to specifically explain the present invention, but the present invention is not limited thereto.

In addition, about the compound synthesize | combined by the parallel method, the structure was confirmed by LC-MS. The measurement conditions are shown below.
[Measurement condition]
LC-MS: Column: INERTSIL ODS 1.5 × 150 mm; Column temperature: 30 ° C .; Measurement wavelength: 220, 254 nm; MS ionization method: APCI method; Eluent A: 0.05% v / v formic acid aqueous solution; 0.05% v / v formic acid acetonitrile solution; flow rate: 0.12 ml / 10 min; gradient conditions: B in A (0-5 min; 5% → 95%, 5-12 min; 95%; 12-14 min , 95% → 5%, 14-25 minutes; 5% (linear gradient)

Reference Example 1 2-amino-benzothiazole-6-carboxylic acid ethyl 4-aminobenzoic acid ethyl ester 16.5 g (0.1 mol), potassium thiocyanate 97.18 g (1 mol), copper sulfate 80 g (0.5 mol), A mixture of 270 mL of methanol was heated and stirred for 3 hours. After standing to cool, insoluble matters were removed by suction filtration. 400 mL of water was added to the filtrate, ethanol was added until the reaction mixture became homogeneous while heating and stirring, and then the reaction mixture was ice-cooled. The precipitated crystals were collected by filtration, washed with water and air-dried to obtain pale yellow crystals. (Yield 11g, Yield 50%)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.34 (d, J = 1.4Hz, 1H), 7.87 (dd, J = 1.4, 8.3Hz, 1H), 7.42 (d, J = 8.3Hz, 1H), 4.30 (q, J = 7.3Hz 2H), 1.32 (t, J = 7.3Hz, 3H).
MS (FAB): 223 (M + 1)

Reference Example 2 2-[(Cyclohexanecarbonyl) amino] -benzothiazole-6-carboxylate Compound 50 g (0.225 mol) of Reference Example 1 and 35 g (0.27 mol) of cyclohexanecarboxylic acid were dissolved in 200 mL of dimethylformamide. 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (hereinafter referred to as EDC) 65 g (0.338 mol), 1-histoxybenzotriazole (hereinafter referred to as HOBt) 46 g (0) .338 mol) was added, and the mixture was stirred at room temperature for 3 hours, and then stirred at 50 ° C. for 16 hours. Crystals poured into 1.5 L of water were collected by filtration, washed thoroughly with water and air-dried. The crystals were washed with hot diisopropyl ether to obtain pale yellow crystals. (Yield 33g, 41%)

¹ H-NMR (CDCl ₃ ) δ (ppm): 10.87 (bs, 1H), 8.57 (d, J = 1.4Hz, 1H), 8.14 (dd, J = 1.4, 8.3Hz, 1H), 7.76 (d, J = 8.3Hz, 1H), 4.43 (q, J = 6.8Hz, 2H), 2.28-2.45 (m, 1H), 1.09-1.95 (m, 13H).
MS (FAB): 333 (M + 1)

Reference Example 3 2-[(Cyclohexanecarbonyl) amino] -benzothiazole-6-carboxylic acid A mixture of 33 g (0.1 mol) of the compound of Reference Example 2 and 1 L of a 1 mol / L aqueous sodium hydroxide solution and 200 mL of methanol was stirred at room temperature for 4 hours. After stirring, methanol was distilled off under reduced pressure. Chloroform was added to the residue for liquid separation, and the aqueous layer was acidified with 10% hydrochloric acid under ice-cooling, and the precipitated crystals were collected by filtration. After thoroughly washing with water and air drying, colorless crystals were obtained. (Yield 25g, 84%)

¹ H-NMR (CDCl ₃ ) δ (ppm): 8.58 (d, J = 1.4Hz, 1H), 7.99 (dd, J = 1.4, 8.3Hz, 1H), 7.77 (d, J = 8.3Hz, 1H) , 3.26 (bs, 1H), 2.48-2.60 (m, 1H), 1.09-1.90 (m, 10H).
MS (FAB): 305 (M + 1)

Reference Example 4 2-[(Cyclohexanecarbonyl) amino] -N-[(2-t-butoxycarbonyl) amino] ethyl) -benzothiazole-6-carboxamide 1 g (3.48 mmol) of the compound of Reference Example 3, N- ( 0.56 g (3.48 mmol) of t-butyl 2-aminoethyl) carbamate was dissolved in 15 mL of dimethylformamide and 1 g of EDC (5.22 mmol) and 0.7 g of HOBt (5.22 mmol) were added to the solution. In addition, the mixture was stirred at room temperature for 16 hours. The reaction solution was poured into water, and the precipitated crystals were collected by filtration, washed thoroughly with water and air-dried. It was subjected to silica gel column chromatography to obtain pale yellow crystals. (Yield 1.28 g, Yield 86%)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 12.34 (s, 1H), 8.42 (s, 1H), 7.90 (d, J = 8.3Hz, 1H), 7.75 (d, J = 8.3Hz, 1H), 6.83 (bs, 1H), 3.25-3.38 (m, 2H), 3.08-3.19 (m, 2H), 2.50-2.62 (m, 1H), 1.16-1.91 (m, 19H).
MS (FAB): 447 (M + 1)

Reference Example 5 2-[(Cyclohexanecarbonyl) amino] -N- (2-aminoethyl) -benzothiazole-6-carboxamide hydrochloride hydrochloride To 210 mg (0.47 mmol) of the compound of Reference Example 4 was added 4 mmol / L-hydrochloric acid / dioxane. 10 mL was added and stirred at room temperature for 16 hours. The crystals were collected by filtration, washed with dioxane and dried under reduced pressure to obtain colorless crystals. (Yield 179 mg, Yield 100%)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 12.38 (bs, 1H), 8.75-8.87 (m, 1H), 8.56 (d, J = 1.4Hz, 1H), 8.05-8.30 (bs, 3H ), 8.00 (dd, J = 8.3, 1.4Hz, 1H), 7.76 (d, J = 8.3Hz, 1H), 7.17 (bs, 2H), 3.48-3.63 (m, 2H), 2.92-3.08 (m, 2H), 2.48-2.65 (m, 1H), 1.13-1.91 (m,
10H).
MS (FAB): 347 (M + 1)

Reference Example 6 N- (2-aminoethyl) -2- (propanoylamino) -benzothiazole-6-carboxamide trifluoroacetate N-[(2-t-butoxy) prepared in the same manner as in Reference Example 4 Carbonyl) amino] ethyl) -2- (propanoylamino) -benzothiazole-6-carboxamide 137 mg (0.35 mmol) was dissolved in 2 mL of dichloromethane, 1 mL of trifluoroacetic acid was added to the solution, and the mixture was stirred at room temperature for 1 hour. . After concentration under reduced pressure and addition of isopropyl ether to the residue for crystallization, the crystals were collected by filtration and dried under reduced pressure to obtain pale yellow crystals. (Yield 90 mg, 63% yield)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 12.50 (s, 1H), 8.65-8.71 (m, 1H), 8.49 (d, J = 1.5Hz, 1H), 7.90-7.95 (m, 5H ), 7.79 (d, J = 8.3Hz, 1H), 3.50-3.58 (m, 2H), 3.00-3.04 (m, 2H), 2.53 (q, 2H, J = 7.3Hz), 1.12 (t, 3H, J = 7.3Hz).
MS (FAB): 293 (M + 1)

  Using the method of Reference Example 4, 5 or 6, Reference Example compounds described in Table 1 were synthesized.

Reference Example 19 2-Amino-6-nitro-benzothiazole 4-Nitroaniline (6.9 g, 0.05 mol) and potassium thiocyanate (9.7 g, 0.1 mol) were dissolved in 70 mL of acetic acid and then ice-cooled. An acetic acid solution containing 2.56 mL (0.05 mol) of bromine was added dropwise to the solution and stirred at room temperature for 16 hours. The reaction solution was poured into water and then neutralized with saturated aqueous sodium hydrogen carbonate. The precipitated crystals were collected by filtration, washed thoroughly with water and air-dried. The crystals were washed with ethyl acetate and then dried under reduced pressure to obtain yellow crystals. (Yield 9.7 g, 99%)

Reference Example 20 N- (6-Nitro-benzothiazol-2-yl) cyclohexanecarboxamide The title compound was obtained by the same procedures as in Reference Example 2 using the compound of Reference Example 19. (Yield 8.5g, Yield 56%)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 12.61 (bs, 1H), 9.09 (d, J = 2.4Hz, 1H), 8.26 (dd, J = 2.4, 8.7Hz, 1H), 7.86 ( d, J = 8.7Hz, 1H), 2.52-2.64 (m, 1H), 1.19-1.95 (m, 10H).
MS (FAB): 306 (M + 1)

Reference Example 21 N- (6-Amino-benzothiazol-2-yl) cyclohexanecarboxamide Using 4g (13.1 mmol) of the compound of Reference Example 20 in 5% palladium-carbon in methanol, catalytic reduction for 2 hours at 4 atm of hydrogen Went. The catalyst was removed by filtration and concentrated under reduced pressure to give the title compound. (Yield 3.5g, Yield 99%)

^{1 H-NMR (DMSO-d} 6) δ (ppm): 11.81 (bs, 1H), 7.38 (d, J = 8.7Hz, 1H), 6.99 (s, 1H), 6.70 (dd, J = 8.7, 1.4 Hz, 1H), 5.06 (bs, 2H), 2.40-2.58 (m, 1H), 1.17-1.83 (m, 10H) .MS (FAB): 276 (M + 1)

Reference Example 22 N-6-{[(3-t-Butoxycarbonylamino) propanoyl] amino} -benzothiazol-2-yl-cyclohexanecarboxamide 330 mg (1.2 mmol) of Reference Example 21 compound, 3-t-butoxycarbonyl The title compound was obtained by the same procedures as in Reference Example 4 using 227 mg (1.2 mmol) of aminopropionic acid. (Yield 321 mg, Yield 60%)

¹ H-NMR (CDCl3) δ (ppm): 10.42 (bs, 1H), 8.29 (s, 1H), 8.15 (s, 1H), 7.63 (d, J = 8.3Hz, 1H), 7.37 (d, J = 6.8Hz, 1H), 5.26 (bs, 1H), 3.48-3.60 (m, 2H), 2.59-2.75 (m, 2H), 2.31-2.43 (m, 1H), 1.16-1.98 (m, 10H), 1.40 (s, 9H).
MS (FAB): 447 (M + 1)

Reference Example 23 N-6-[(3-Aminopropanoyl) amino] -benzothiazol-2-yl-cyclohexanecarboxamide hydrochloride hydrochloride The same method as Reference Example 5 using 325 mg (0.73 mmol) of the compound of Reference Example 22 Gave the title compound as colorless crystals. (Yield 270 mg, 99%)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 12.14 (bs, 1H), 10.44 (s, 1H), 8.29 (d, J = 1.9Hz, 1H), 8.01 (bs, 2H), 7.65 ( d, J = 8.3Hz, 1H), 7.55 (dd, J = 8.3, 1.9Hz, 1H), 6.30 (bs, 2H), 3.02-3.15 (m, 2H), 2.71-2.81 (m, 2H), 2.41 -2.59 (m, 1H), 1.13-1.83 (m, 10H).
MS (FAB): 347 (M + 1)

Reference Example 24 2-[(Naphthalen-2-ylmethyl) amino] ethyl-carbamate-t-butyl N- (2-aminoethyl) carbamate t-butyl 8.65 g (54 mmol), 2-naphthaldehyde 7.03 g (45 mmol) was dissolved in 180 mL of dichloroethane, 14.3 g (67.5 mmol) of sodium triacetoxyborohydride was added to the solution, and the mixture was stirred at room temperature for 4 hours, and then saturated aqueous sodium hydrogen carbonate was added to the reaction solution to stop the reaction. After separating the organic layer, the aqueous layer was extracted with chloroform. The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The residue was subjected to silica gel column chromatography to give the title compound as a pale yellow oil. (Yield 10.26 g, Yield 76%)

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.80-7.87 (3H, m), 7.74 (1H, s), 4.95 (1H, br), 3.95 (2H, s), 3.22-3.32 (2H, m ), 2.78-2.83 (2H, m), 1.44 (9H, s).
MS (FAB): 301 (M + 1)

Reference Example 25 (2-t-Butoxycarbonylaminoethyl) -naphthalen-2-ylmethyl-allylcarbamate 23.2 g (77.33 mmol) of the compound of Reference Example 24 was dissolved in 140 mL of tetrahydrofuran. An aqueous solution (80 mL) of 21.34 g (154.66 mmol) of potassium carbonate was added to the solution and cooled with ice. To the reaction solution, 9.55 g (79.27 mmol) of allyl chlorocarbonate was added dropwise, and then stirred at room temperature for 2 hours. The reaction mixture was diluted with water, extracted with ethyl acetate, and washed with saturated brine. The extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography to obtain a pale yellow oil. (Yield 29.7 g, 100%)

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.78-7.84 (m, 3H), 7.60-7.73 (m, 1H), 7.26-7.50 (m, 3H), 5.91-6.02 (m, 1H), 5.22 -5.36 (m, 2H), 4.69 (s, 4H), 3.27-3.43 (m, 4H), 1.42 (s, 9H).
MS (FAB): 385 (M + 1)

Reference Example 26 (2-aminoethyl) -naphthalen-2-ylmethyl-allylcarbamate 29 g (77 mmol) of the compound of Reference Example 25 was dissolved in 100 mL of dichloromethane, 100 mL of trifluoroacetic acid was added to the solution, and the mixture was stirred at room temperature for 2 hours. Stir. The reaction mixture was concentrated under reduced pressure, 100 mL of dichloromethane was added to the residue, and the mixture was again concentrated under reduced pressure. The residue was dissolved in chloroform, washed successively with saturated aqueous sodium hydrogen carbonate and saturated brine, and dried over anhydrous potassium carbonate. After concentration under reduced pressure, the residue was subjected to alumina column chromatography to give the title compound as a pale yellow oil. (Yield 16.27 g, 74% yield)

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.77-7.86 (m, 3H), 7.61-7.74 (m, 1H), 7.29-7.55 (m, 3H), 5.83-6.03 (m, 1H), 5.16 -5.40 (m, 2H), 4.68 (s, 4H), 4.25 (brs, 2H) 3.39-3.54 (m, 2H), 2.91-3.07 (m, 2H).
MS (FAB): 285 (M + 1)

Reference Example 27 N- {2-[(2-Nt-butoxycarbonylamino-benzothiazol-6-yl) carbonyl] -aminoethyl} -N- (2-naphthylmethyl) carbamate allyl compound of Reference Example 26 7.1 g (25 mmol), 2-Nt-butoxycarbonylamino-benzothiazole-6-carboxylic acid 6.85 g (25 mmol) in dimethylformamide solution (100 mL) 7.17 g (37.5 mmol) EDC, HOBt 5.07 g (37.5 mmol) was added and stirred at room temperature for 16 hours. The reaction solution was poured into 500 mL of water, and the precipitated crystals were collected by filtration and washed thoroughly with water. After air drying, the title compound was obtained as colorless crystals by silica gel column chromatography. (Yield 11.2 g, Yield 80%)

¹ H-NMR (CDCl ₃ ) δ (ppm): 8.26 (s, 1H), 7.77-7.89 (m, 5H), 7.62-7.71 (m, 1H), 7.32-7.49 (m, 3H), 5.88-5.98 (m, 1H), 5.14-5.33 (m, 2H), 4.64-4.75 (m, 4H), 3.54-3.68
(m, 4H), 1.61 (s, 9H).
MS (FAB): 561 (M + 1)

Reference Example 28 N- {2-[(2-amino-benzothiazol-6-yl) carbonyl] -aminoethyl} -N- (2-naphthylmethyl) carbamate allyl compound of Reference Example 27 5.5 g (9. 8 mmol) was dissolved in 20 mL of dichloromethane, 10 mL of trifluoroacetic acid was added to the solution, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in a small amount of chloroform, and saturated aqueous sodium hydrogen carbonate was added to the solution until basic. The precipitated crystals were collected by filtration, washed with chloroform and dried under reduced pressure to give the title compound as colorless crystals. (Yield 4.5g, 99%)

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.10 (s, 1H), 7.82-7.91 (m, 3H), 7.66-7.78 (m, 3H), 7.30-7.53 (m, 3H), 5.89 -5.97 (m, 1H), 5.10-5.36 (m, 2H), 4.54-4.71 (m, 4H), 3.37-3.49 (m, 4H).
MS (FAB): 461 (M + 1)

Reference Example 29 {2-[(2-acetylamino-benzothiazole-6-carbonyl) -amino] -ethyl} -naphthalen-2-ylmethyl-allylcarbamate Compound 138 mg (0.3 mmol) 4-dimethyl of Reference Example 28 To a mixture of 37 mg (0.3 mmol) of aminopyridine and 10 mL of dichloromethane, 92 mg (0.9 mmol) of acetic anhydride was added and stirred at room temperature for 4 hours. The reaction solution was washed with 3 mmol / L-hydrochloric acid and then washed successively with water and saturated brine. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography to obtain the title compound as colorless crystals. (Yield 137 mg, Yield 91%)

¹ H-NMR (CDCl ₃ ) δ (ppm): 8.30 (s, 1H), 7.35-7.84 (m, 9H), 5.86-5.97 (m, 1H), 5.15-5.34 (m, 2H), 4.66-4.75 (m, 4H), 3.62-3.69 (m, 4H), 2.30 (s, 3H).
MS (FAB): 503 (M + 1)

  Using the method of Reference Example 23, Reference Example compounds described in Tables 2 and 3 were synthesized.

Example 1 2-[(Cyclohexanecarbonyl) amino] -N-2-[(2-naphthylmethyl) amino] ethyl-benzothiazole-6-carboxamide Hydrochloride of Reference Example 5 (115 mg, 0.3 mmol), 2- To a mixture of naphthaldehyde (25 mg, 0.24 mmol) and dimethylformamide 2 mL, 430 mg (0.9 mmol) of MP-triacetoxyborohydride (Argonaute Technology) as a solid-phase-supported reagent was added and stirred at room temperature for 12 hours. did. PS-benzaldehyde (manufactured by Argonaute Technology) corresponding to 0.2 mmol was added to the reaction solution and further stirred for 6 hours. The reaction solution was filtered, and once adsorbed on a cation exchange resin MP-TsOH cartridge (manufactured by Argonaute Technology), washed sequentially with dimethylformamide, dichloromethane and methanol, and then eluted with 2M ammonia / methanol. A highly pure crude product was obtained by concentrating the ammonia / methanol solution under reduced pressure. The crude product was subjected to 1 g of silica gel column chromatography to obtain 43 mg (yield 37%) of the title compound.

¹ H-NMR (CDCl ₃ ) δ (ppm): 8.26 (d, J = 1.4Hz, 1H), 7.75-7.85 (m, 5H), 7.71 (d, J = 8.2Hz, 1H), 7.41-7.47 ( m, 3H), 6.95-7.04 (m, 1H), 4.02 (s, 2H), 3.58-3.65 (m, 2H),
2.95-3.05 (m, 2H), 2.35-2.45 (m, 1H), 1.18-2.01 (m, 10H).
LC-MS (APCI): 487 (M + 1)
LC-Retention time 9.05 min.

  The parallel synthesis which adapted the method similar to Example 1 to the corresponding amine and aldehyde was performed, and the compound of Examples 2-167 described in Tables 4-16 was synthesized. All compounds were confirmed to have a purity of 95% or more by LC-MS.

Example 168 N- [6- (3-[(2-naphthylmethyl) amino] propanoylamino) -benzothiazol-2-yl] cyclohexanecarboxamide Example 1 using the compound of Reference Example 23, 2-naphthaldehyde The title compound was obtained by carrying out the reaction according to the method described.

¹ H-NMR (CDCl ₃ ) δ (ppm): 10.70 (s, 1H), 8.27 (d, J = 1.4Hz, 1H), 7.87-7.90 (m, 4H), 7.63 (d, J = 8.3Hz, 1H), 7.42-7.55 (m, 3H), 7.35 (d, J = 8.3Hz, 1H), 4.06 (s, 2H), 3.08-3.15 (m, 2H), 2.54-2.60 (m, 2H), 2.30 -2.39 (m, 1H), 1.20-1.97 (m, 10H).
MS (FAB): 487 (M + 1)

  The same procedure as in Example 168 was performed using the corresponding amine and aldehyde, and the compounds of Examples 168 to 219 described in Table 17 to Table 22 were synthesized.

Example 220 2- (acetylamino) -N-2-[(2-naphthylmethyl) amino] ethyl-benzothiazole-6-carboxamide 50 mg (0.1 mmol) of the compound of Reference Example 29, 6 mg of tetrakistriphenylphosphine palladium ( 0.005 mmol), 1 mL of anhydrous tetrahydrofuran was added to 100 mg (0.1 mmol) of solid-supported barbituric acid, and the mixture was stirred at 50 ° C. for 12 hours under an argon stream. The reaction solution was filtered, the polystyrene resin was washed with tetrahydrofuran, and the filtrate was concentrated under reduced pressure. The residue was subjected to 1 g of silica gel column chromatography to obtain the title compound as colorless crystals. (Yield 33 mg, 79% yield)

¹ H-NMR (CDCl ₃ ) δ (ppm): 8.24 (s, 1H), 7.65-7.84 (m, 6H), 7.43-7.52 (m, 3H), 3.98-4.04 (m, 2H), 3.56-3.63 (m, 2H), 2.93-3.01 (m, 2H), 2.28 (s, 3H)
MS (FAB): 419 (M + 1)

  Using the intermediate prepared by the same method as in Reference Example 29, the reaction was carried out by the method described in Example 220 to obtain the compounds of Examples 221 to 240 described in Tables 23 and 24.

Example 241
The antifungal effect was measured using the following test methods.
1. Candida albicans and H.C. sapiens N-myristoyltransferase inhibition test 1-1. Candida albicans and H.C. Preparation of recombinant N-myristoyltransferase of sapiens Recombinant enzyme was prepared by a known method (Rajala VS; Raju SS; Rajendra KS; Protein Expr. Purif. 1996, 7). , 431).
Candida albicans or H.C. The sapiens N-myristoyltransferase gene was inserted into an expression vector for Escherichia coli pET-15b (Novagen: 69661-3) to construct each expression vector. After protein expression by E. coli, E. coli was disrupted and each N-myristoyltransferase was obtained by metal affinity chromatography.

1-2. Candida albicans and H.C. sapiens N-Myristoyltransferase Inhibition Test The enzyme inhibition test was performed as follows based on a known method (see Dlight t .; Luis g .; PNAS 1986, 83, 2812).
Test compound solution: A test compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a dilution series as appropriate from a maximum of 10 ⁻⁴ mol / L of 1% DMSO solution and added to the reaction solution.
Enzyme inhibition test: Reaction buffer (10 mM Tris-HCl, pH 7.4 / 1 mmol / L dithiothreitol / 5 mmol / L MgCl ₂ /0.1 mmol / L
During EGTA / 0.01% TritonX-100) 100μL, [3 H] Myr-CoA (7.4kBq / final concentration 35.7nmol / L), substrate peptide (final concentration 10μmol / L), and test various concentrations After adding the compound and preincubating for 10 minutes at 30 ° C., Candida albicans The enzyme reaction was performed by adding sapiens recombinant enzyme. After reacting at 30 ° C. for 10 to 20 minutes, 100 μL of methanol containing 10% TCA was added to stop the reaction, and the reaction tube was left on an ice bath. After fractionating the reaction product from the reaction solution by HPLC, the reaction product was quantified with a liquid scintillation counter, and the 50% inhibition concentration IC ₅₀ of the test compound was calculated from the inhibition rate at various concentrations of the test compound.

2. Measurement of antifungal activity Measurement of in vitro antifungal activity against Candida albicans The following operation was carried out according to NCCLS protocol M27-A2.
Test compound solution: The test compound was dissolved in dimethyl sulfoxide (DMSO) to obtain a solution of maximum 3.2 mg / mL, and diluted with DMSO to prepare a 2-fold dilution series. These solutions were added to the test medium at a final concentration of 1% (V / V).
Test medium: After 10.44 g of RPMI 1640 was dissolved in 900 mL of distilled water, MOP buffer (0.165 mol / L) was added and dissolved. Next, after adjusting the pH to 7.0 with a 5 mol / L sodium hydroxide aqueous solution, distilled water was added to make 1000 mL, and the solution was sterilized by filtration.
Inoculum: C.I. albicans ATCC 90028 and C.I. After culturing kruse ATCC 6258 in Sabouraud dextrose agar medium at 35 ° C. for 24 hours, subcultured once more under the same conditions, collecting 5 colonies with a diameter of 1 mm or more and suspending in 5 mL of sterilized physiological saline did. The absorbance of this suspension was adjusted to McFarland 0.5, and then diluted 1000 times with a test medium to obtain an inoculum solution.
Antifungal activity measurement: 100 μL of each diluted solution of the specimen in the test medium was dispensed into each well of a 96-well round bottom microplate, and 100 μL of the above inoculum was added (final bacterial concentration: 0.5-2. 5 × 10 ³ cells / mL) and cultured at 35 ° C. for 48 hours. After completion of the culture, the absorbance at 600 nm was measured, and the minimum drug concentration at which the growth of the bacteria was suppressed by 80% or more compared with the drug-free control was defined as MIC] (μg / mL).

The calcium N-myristoyltransferase inhibitory action (IC ₅₉ value) of the benzothiazole derivative of the present invention was generally in the range of 0.02 to 100 μmol / L. On the other hand, it was confirmed that the action on human N-myristoyltransferase (NMT) was very weak and the fungal selectivity was high. The measurement results are shown in Table 25.

  C. albicans ATCC 90028 and C.I. Table 26 shows the measurement results of antifungal activity against krusei ATCC 6258.

Claims (3)

The following general formula (I)

(Wherein R ¹ is a cycloalkyl group which may have a substituent aromatic hydrocarbon group or an optionally substituted nitrogen atom, an aromatic heterocyclic group containing an oxygen atom or a sulfur atom R ² has a lower alkyl group, an aralkyl group which may have a substituent, a cycloalkyl group, an oxiranyl group, an aromatic hydrocarbon group which may have a substituent or a substituent. Or an aromatic heterocyclic group containing a nitrogen atom, oxygen atom or sulfur atom, wherein X represents a group —Y—CH (R ³ ) —Z—N (R ⁴ ) —CO— or a group —Z—CONH—. Y represents a group — (CH ₂ ) _n — (where n represents 0 or 1), Z represents a linear or branched lower alkylene group, or a 5- to 6-membered cycloalkyl group. , R ³ and R ⁴ each independently represent a hydrogen atom, or a lower alkyl group, or R ³ and R ⁴ Shows the saturated heterocyclic group of 4 to 6-membered ring containing a nitrogen atom together)
Or a pharmaceutically acceptable salt thereof.   A pharmaceutical comprising the 2-aminobenzothiazole derivative represented by the general formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.   The medicament according to claim 2, which is an antifungal agent.