PL151588B1 - The method of manufacture of new derivatives of azoles - Google Patents

Description

RZECZPOSPOLITA PATENT DESCRIPTION 151 588

POLAND '

OFFICE

PATENT

RP

Additional patent to patent no. - Filed: 87 12 02 (P. 269 180)

Int. Cl. ⁵ C07D 233/61 C07D 249/08

Priority: 86 12 03 Federal Republic of Germany tintiiu

ICKU

Application announced: 88 08 18

Patent description published: 1991 04 30

Inventor: - Authorized by the patent: Sandoz AG,

Basel (Switzerland)

A method of producing new azole derivatives

The present invention relates to a process for the preparation of new azole derivatives which are suitable for the production of pharmaceuticals.

The process of the invention produces azole derivatives of the formula I, as shown in the figure, in which each of R1 and R2 is an unsubstituted or monosubstituted phenyl group with a halogen atom, R3 is a C1-6alkyl, C1-6 alkoxy, indolyl, indanyl, C1-4alkyl group substituted cyano, benzyloxy or polyether group, phenyl group monosubstituted with halogen atom or cyano group or biphenylyl group monosubstituted with halogen atom and X is CH or N, whereby when R3 is methyl group and R1 and R2 are phenyl group then at least one phenyl group must be substituted.

Halogen atoms, such as chlorine, bromine, iodine or fluorine, especially fluorine or chlorine, are present as substituents in the groups R1 and R2. Monosubstitution is preferred, especially in the para position when R1 and R2 are phenyl.

Preferred substituents in the R3 group are halogen atoms, e.g., fluorine or chlorine, or cyano.

R3 as alkyl preferably has 1 to 4 carbon atoms and may be substituted by cyano, alkoxy, benzyloxy or polyether (e.g. alkoxy / mono-, di- or triethyleneoxy). .

Lower alkyl groups which are themselves substituents or are included in the substituents preferably contain 1 to 4 carbon atoms. '

The following meanings of the symbols in formula 1 are preferred: R1 and R2 are pF- especially p-Cl-phenyl, X is CH, R3 has the preferred meanings defined above, especially ρ-F-, p-Cl- or p-CN- phenyl.

In a particular group of compounds of formula I, R 1, R 2 and X are as defined above and R 3 is unsubstituted or monosubstituted alkyl or phenyl.

Compounds of formula I may exist in free form or in acid addition salt form.

151 588

By the process of the invention, compounds of formula 1 in free form or in acid addition salt form are obtained by acylating a compound of formula 2 wherein R 1, R 2 and X are as defined above, and recovering the resulting compounds in free form or in addition salt form with acid.

The acylation is preferably carried out using a carboxylic acid R3COOH in which _R3 is as defined above, or a reactive derivative thereof by a conventional method.

The process of the invention can be carried out, for example, by reacting a compound of formula 2 with the appropriate acid halide in a solvent which is inert under the reaction conditions, e.g. an essentially organic or inorganic base which simultaneously functions as an acid binder such as pyridine, optionally by adding an acylation accelerator such as 4-dimethylaminopyridine. For acylation, a compound of formula 2 can also be reacted with an active ester, for example the acid R3COOH. The reaction may be performed, for example, with a pyridyl-2-thiol ester in a solvent which is inert under the reaction conditions, e.g. in a di-lower alkyl carboxylic acid amide such as dimethylformamide, at room temperature. Furthermore, a compound of formula 2 can be reacted with R3COOH in the presence of N-hydroxybenzotriazole and dicyclohexylcarbodiimide. The final product can be isolated from the reaction mixture by known methods and purified if necessary.

The starting compounds of formula II are partly novel and can be prepared, for example, according to the reaction scheme shown in the figure. The remaining starting compounds, as well as the acylating agents, are known or can be prepared analogously to known methods or correspondingly analogously to the methods described in the examples.

The compounds of formula I exhibit pharmacological activity and are therefore indicated for use as pharmaceuticals.

The compounds exhibit aromatase inhibitory activity in the following tests.

1. In vitro aromatase inhibition. Human placenta microsomes were used as the source of the enzymes. To obtain these microsomes, the entire human placenta was transferred to the laboratory at low temperature immediately after birth, excess blood was removed by intensive washing with 0.25 M sucrose, then the placenta was cut into pieces and immediately homogenized by cooling (1 g organ + 1.5 g KCl / tris buffer , pH 7.4; ultraturrax 3X 10s / 0 ° C). After sedimentation of partially disrupted cells, nuclei and mitochondria at 200, 7000 and 11000 g for 10 minutes, a microsome fraction was obtained by centrifuging the post-mitochondrial supernantant at 105,000 g / 60 min. The pellet, which was washed, was resuspended in isotonic KC1 solution with tris buffer to obtain a Cyt P-450 concentration of 0.5-1 µΜ.

Aromatase activity is determined by a modified known method (Thompson FA, Siiteri PK / 1974 /, J. Biol. Chem. 249, 5364-5372) using the L / L, 2 / J ³ H-androstenedione as the substrate in the presence of NADPH regenerating system. Test compounds are added at a concentration of 0.01 and 1 µΜ. After incubation for 30 minutes at 37 ° C, the reaction is terminated by mixing with a large excess of chloroform and the aqueous and organic phases are separated. The radioactivity of the aqueous phase is assigned a ³ HAO, which is formed during the aromatization of l / I, 2 / L ³ H-androstenedione to estrone and used as an indicator of enzymatic activity. Compounds that significantly inhibit aromatase activity under these assay conditions are then tested over a wide range of concentrations to determine IC50 values.

2. Inhibition of ovulation. Female rats with regular cycles received the test substance at 4 p.m. the day before ovulation and at 11:00 on the day of ovulation. Ovulation was determined by counting the eggs in the vaginal fluid.

3. Effects on ovarian aromatase and E2 production in vivo. Adult female rats were initially dosed subcutaneously for 12 days with 25IU (daily PMSG / gonadotropin from pregnant mares plasma). On day 13, the test substance was orally administered at a concentration of 0.032 to 10 mg / kg. The rats were then sacrificed 8, 24 and 48 hours later and the blood and ovaries were removed. Microsomes were obtained from one appropriate ovary of each animal, and aromatase activity was determined as described above. Estradiol was extracted from the second ovary and quantified by RIA; blood estradiol was similarly determined.

4. Subchronic action in vivo. The test substance was administered once a day for 7 days to young, 23-day-old female rats. From the 4th to the 7th course, testosterone propionate was additionally injected subcutaneously. On the 8th day the animals were killed. Blood was collected, the uterus was removed

151 588 cleaned and weighed. The administration of testosterone causes a significant increase in the weight of the uterus, which is caused by the conversion of testosterone to estradiol by aromatase. Thus, the inhibition of uterine hyperplasia by the test substance is an accurate measure of the inhibition of estrogen production by aromatase during the 7 days of administration.

5. Chronic action. In rats, estrogen-dependent mammary tumors were induced by single oral administration of 7,12-dimethylbenz [a] anthrazene (DMBA). Treatment with test substance was initiated when tumors averaged approximately 1500 mm ³ in size and continued for up to 6 weeks.

The compounds of formula I exhibit this activity when administered once or twice daily orally at concentrations from about 1 mg / kg to about 10 mg / kg.

Aromatase plays an important role in the biosynthesis of estrogen in mammals. Blocking this enzyme by specific inhibitors can drastically lower the normal level of steroids. Such non-invasive chemical reduction of steroid levels, as opposed to the surgical removal of hormone-producing organs (prostate, ovary, adrenal gland), appears as a desirable alternative and complement to the treatment of steroid-dependent tumors, e.g. prostate, polycystic ovary, cancer of the breast, ovaries, intrauterine, as well as Cushing's syndrome.

The compounds of formula I are therefore indicated for use as specific inhibitors of steroid synthesis for the treatment of tumors as mentioned above. An appropriate daily dosage as appropriate is in the range of from about 10 mg to about 100 mg of the compound, conveniently administered, e.g. in divided doses up to 4 times daily.

The compounds of formula I can be administered by conventional means, especially enterally, preferably orally, e.g. in the form of tablets or capsules, or parenterally, e.g. in the form of injection solutions or suspensions.

Compounds of formula 1 can be administered as the free base or as a pharmaceutically acceptable acid addition salt. Such salts can be prepared in a conventional manner and exhibit the same order of activity as the free bases. By conventional method, pharmaceutical compositions can be prepared which contain a compound of Formula 1 as the free base or in the form of a pharmaceutically acceptable acid addition salt together with at least one pharmaceutical carrier or diluent. Unit dosage form contains, for example, from about 2.5 mg to about 50 mg of a compound of Formula 1 as the free base or as a pharmaceutically acceptable acid addition salt.

The compounds of formula I, prepared by the process of the invention in free base form or in the form of pharmaceutically acceptable acid addition salts, are useful as pharmaceuticals, in particular as aromatase inhibitors, and are suitable for the preparation of medicaments intended to inhibit aromatase.

In the following examples, which more fully illustrate the invention, all temperatures are given in degrees Celsius in uncorrected form.

Example I. Preparation of 2,2-bis- (4-chlorophenyl) -2- (1H-imidazol-1-yl) -1- (4-chlorobenzoylamino) ethane.

0.34 ml of 4-chlorobenzoyl chloride was added dropwise with ice cooling to 670 mg of 2,2-bis- (4-chlorophenyl) -2- (1H-imidazol-1-yl) -1-aminoethane in 4 ml of dry pyridine. The mixture was stirred for 4 hours at room temperature. It was then poured into ice, saturated with sodium bicarbonate solution and extracted with ethyl acetate. The combined organic phases are washed with saturated sodium hydrogen carbonate solution and with sodium chloride solution, dried over magnesium sulfate and concentrated by evaporation. The residue was recrystallized after being dissolved warm in ethyl acetate and diethyl ether. Colorless crystals with a melting point of 241-244 ° were obtained.

2,2-Bis- (4-chlorophenyl) -2- (1H-imidazol-1-yl) -1-aminoethane used as a starting compound can be prepared as follows:

a) 2,2-bis- (4-chlorophenyl) aziridine.

A Grignard solution consisting of 8.45 g of magnesium and 36 ml of bromobenzene in 200 ml of absolute ether was mixed with 300 ml of dry toluene and the ether was distilled off until an internal temperature of 107 ° was reached. Then, 28.8 g of p-chloroacetophenoneoxime in 100 ml of dry toluene was added dropwise to the boiling solution, the mixture was heated under reflux for 1.5

For 151,588 hours, it was mixed with 300 ml of 20% ammonium chloride solution under ice cooling. After phase separation, the aqueous phase was extracted with ether. The combined organic phases are washed with saturated sodium chloride solution, dried over magnesium sulfate and concentrated by evaporation. The oily residue was chromatographed on silica gel (0.040 to 0.063 mm) eluting with 60-80 ° petroleum ether and diethyl ether / petroleum ether (3/2). The brown oil thus obtained was characterized by the NMR spectrum. ¹ H-NMR (CDCl 3): 2.36 (2H, s, broad, -CH2); 7.20-7.55 (8H, m, aromat).

b) 2,2-Bis- (4-chlorophenyl) -2- (1H-imidazol-1-yl) -1-aminoethane.

3.96 g of 2,2-di- (4-chlorophenyl) aziridine was heated to 95 ° (until melted) for 20 hours with

5.1 g of imidazole. Ethyl acetate / water solvents were added to the melt and the aqueous phase was extracted several times with ethyl acetate. The organic phases are washed with 0.25% tartaric acid solution and saturated sodium chloride solution, dried over magnesium sulfate and concentrated by evaporation. A residue which, in addition to the desired product, also contains a positional isomer

1.1-bis- (4-chlorophenyl) -2- (1H-imidazol-1-yl) -1-aminoethane, partitioned on silica gel (0.040-0.063 mm) using solvents dichloromethane / methanol / petroleum ether (20/1 / 5). A sticky yellow-brown oil was obtained. ¹ H-NMR (CDCl3): 1.40 (2H, broad, -NH2); 3.90 (2H, s, -CH2); 6.9-7.5 (6H, m, aromat, imidazole); 7.68 (1H, imidazole).

The following starting materials can be obtained analogously: 2,2-bis-phenyl-2- (1H-imidazol-yl) -1-aminoethane (for Example 2) obtained as a viscous yellowish oil, 2,2-bis- (4-chlorophenyl) - 2- (1H-1,2,4-triazol-1-yl) -1-aminoethane (for Example 3) obtained as sticky yellow-brown oil; ¹ H-NMR (CDCl3): 1.75 (2H, s, broad, -NH2); 3.93 (2H, s, -CH2); 6.9-7.5 (8H, m, aromat); 7.88 (1H, s, triazole); 8.10 (1H, s, triazole).

The following compounds of formula 1 can be obtained analogously to that described in Example 1 starting from the corresponding starting materials.

Table

Ex. X R1 R ₂ Ra T. m.p. ° C II CH c ₆ h ₅ C ₆ H ₅ pattern 3 193-197 ° III N pattern 3 pattern 3 pattern 3 158-162 ° IV CH pattern 3 pattern 3 -ch ₃ 178-182 ° V CH pattern 3 pattern 3 -C (CH ₃ ) 3 131-136 ° VI CH pattern 3 pattern 3 pattern 4 220-224 ° VII CH pattern 3 pattern 3 pattern 5 144-146 ° VIII CH pattern 3 pattern 3 pattern 6 amorph. IX CH pattern 3 pattern 3 OC ₂ H ₅ amorph. X CH pattern 3 pattern 3 pattern 7 amorph. (racemate) XI CH pattern 3 pattern 3 pattern 6 253-256 ° XII CH pattern 3 pattern 3 formula 8 233-235 ° XIII CH pattern 3 pattern 3 CH ₂ (O-CH ₂ -CH ₂ ) 3OCH3 Oil XIV CH pattern 3 pattern 3 ch ₂ -cn 219-220 ° XV CH pattern 9 pattern 9 pattern 9 218-220 ° XVI CH pattern 9 pattern 9 pattern 3 229-232 ° XVII CH pattern 9 pattern 9 ch ₃ 181-183 °

Claims (1)

Patent claim A process for the preparation of new azole derivatives of formula I, wherein each of R1 and R2 is an unsubstituted or monosubstituted phenyl group, R3 is a C1-6alkyl, C1-4alkoxy, indolyl, indanyl, C1-4alkyl substituted cyano, benzyloxy or polyether, phenyl group monosubstituted with an atom 151 588 halogen or a cyano group or a biphenylyl group monosubstituted with a halogen atom and X is CH or N, whereby when R3 is a methyl group and Ri and R2 are a phenyl group, then at least one of these phenyl groups must be substituted in the form of free or in the form of an acid addition salt, characterized in that a compound of formula II is acylated in which R1, R2, and X are as defined above with a compound of formula R3COOH in which R3 is as defined above or a reactive derivative thereof and the resulting compound is recovered in free form or in acid addition salt form. Formula 5 R ^ C-CHj-NH-COr ₂ Formula 1 Rl-C- ^CH 2- NH ₂ Formula 2 Formula 6 Cl Formula 8 CN Formula 3 Pattern Cl Wżór 9 Formula 4 151 588 R ₁ -CO-CH ₃ ♦ NH ₂ OH.HC1 - ^ • R ₁ -C-CH ₃ + 2 R ₂ MgHai Formula 2 Scheme Department of Publishing of the UP RP. Circulation 100 copies Price: PLN 3,000