SE444934B - ALFA-DIFLUORMETHYL DERIVATIVES OF ALFA-AMINO ACIDS AS A PROCEDURE FOR PRODUCING THEREOF - Google Patents

Description

7807691-6 Group IIIA metals, for example aluminum, organic amines, for example primary, secondary or tertiary amines, for example cyclohexylamine, ethylamine, pyridine, methylaminoethanol, ethanolamine and piperazine. The salts are prepared in a conventional manner. Illustrative examples of compounds of the present invention are the following = 2-Amino-2-difluoromethyl-5-guanidinovaleric acid (Formula I) 2,5-diamino-2-difluoromethylvaleric acid (Formula II) methyl-2,5-diamino-2 -difluoromethylvalerate.

Compounds of general formula I can be used in many ways.

The compounds of the general formula I and II are inhibitors of decarboxylase enzymes which are involved in polyamine formation, which makes said compounds useful as pharmacological agents. Polyamines, especially putrecin, spermicin and spermine are present in plants and animal tissues and in certain microorganisms. Although the exact physiological role of polyamines is not clear, there is evidence to suggest that polyamines are involved in cell division and growth (Williams-Ashman et al., The Italian J. Biochem. 25, 5-32 (1976) Raina and Janne , Med. Biol. 53, 121-147 (1975) and Russell, Life Science 13, 1635-1647 (1973)). The polyamines are essential growth factors involved in the growth processes of certain microorganisms, such as E. coli, Enterobacter, Klebsiella, Staphylococcus aureus, C. cadaveris, Salmonella typhosa and Haemophilus parainfluenza. The polyamines are associated with both normal and neoplastic rapid growth, with an increase in the synthesis and accumulation of polyamines after a stimulus-causing cell growth. The level of polyamines is also known to be high in embryonic systems, in samples from patients with rapidly growing tissue. It is known that there is an association between the activity of decarboxylase enzymes of ornithine, S-adenosylmethionine, arginine and lysine and polyamine formation; The biosynthesis of putrescine, spermidine and spermine are related to each other. Putrescine is the decarboxylation product of ornithine. tin, catalyzed by ornithine decarboxylase. Putrescine formation can also occur by decarboxylation of arginine to form agmatine, which is hydrolyzed to obtain putrescine and urea. Arginine is also involved in ornithine formation through the action of enzyme arginase. Activation of methionine by S-adenosylmethionine synthetase forms S-adenosylmethionine, which is decarboxylated, after which the propylamine portion of activated methionine can be converted to putrescine to form spermidine or the polyamine can be transferred to spermidine to form spermine. Thus, putrescine serves as a precursor to spermidine and spermine and furthermore, it has been shown to have a marked regulatory effect on the polyamine biosynthetic pathway in that it has been shown to increase the synthesis of putrescine, which is the first indication that a tissue will undergo - new growth. Cadaverine, which is the decarboxylation product of lysine, has been shown to stimulate the activity of S-adenosylmethionine decarboxylase and is known to be essential for the growth of many microorganisms, e.g. H. parainfluenza.

The compounds of the general formula II, in which Z represents HZN (CH2) 3-oxylase and in which Z represents Y-guanidinopropyl are inhibitors of and the lactams thereof, are inhibitors of ornithine carbargin-decarboxylase. As inhibitors of the above-mentioned decarboxylase enzymes, the compounds of general formula I and II are useful as anti-infective agents for regulating microorganisms, for example bacteria and viruses, which depend on polyamines for growth, for example E. coli, Enterobacter, Klebsiella , Staphylococcus aureus, C. cadaveris, viruses such as H. parainfluensa, picornavirus, for example encephalomyocarditis, Herpes simplex, syphilis virus and arbovirus, for example Semliki forest. The compounds of the general formula I and II are also useful for regulating certain rapid growth processes. For example, the compounds can be used to inhibit spermatogenesis and embryogenesis and therefore the compounds find use as male antifertility agents and abortifacients. These compounds are also useful for inhibiting the immune response and thus the compounds can be used as suppressive agents for the immune response and for regulating neoplastic growth, for example solid tumors, leukemia and lymphomas. The compounds are also useful as inhibitors of prostatic hypertrophy, excessive scalp cell growth, as evidenced by the presence of dandruff, and as an inhibitor of abnormal skin cell growth manifested in psoriatic conditions. The use of the compounds of general formula I and II as irreversible inhibitors of ornithine or S-adenosylmethionine decarboxylase enzymes in vivo can be shown in the following manner. An aqueous solution of a suitable compound of formula I or II is administered orally or parenterally to male mice or rats. The animals are killed 1-48 hours after administration of the compound and the abdominal flaps of the prostate are removed and homogenized, measuring the activity of ornithine or S-adenosylmethionine decarboxylase enzyme as generally described by Pegg and Williams-Ashman, Biochem. J. 108, 533-539 (1968) and Janne and Williams-Ashman, Biochem. and Bio- phys. Res. Comm. 42, 222-228 (1971).

The compounds of general formula I and II are useful as chemical intermediates for the preparation of novel cephalosporin derivatives which are useful as antibiotics and have the following general structure: cr 'H. 2 S z-c-coNH f IÉIHZ á L_ /._CH2X '' Formula III cooM where Z is previously defined, M represents hydrogen or a negative charge and X represents hydrogen or a negative charge and X represents hydrogen or acetoxy.

The compounds of general formula III and pharmacologically acceptable salts and individual optical isomers thereof are novel chemical compounds useful as antibiotics, which can be administered in a manner similar to many well-known cephalosporin derivatives, for example cefalexin, cephalothin or cephaloglycin. The compounds of the general formula III and pharmacologically acceptable salts and isomers thereof can be administered alone or in the form of pharmacological preparations either orally or parenterally and topically to warm-blooded animals, i.e. birds and mammals, for example cats, dogs, cattle, sheep, horses and to people. For oral administration the compounds may be administered in the form of tablets, capsules or pills or in the form of elixirs or suspensions, for parenteral administration the compound may be used in the form of sterile aqueous solutions which may contain other solutes, e.g. too much salt or glucose to make the solution isotonic. For topical administration of the compounds of general formula III, salts or isomers thereof may be incorporated into creams or liniments.

Illustrative examples of bacteria against which the compounds of formula III and pharmacologically acceptable salts and individual optical isomers thereof are active are Staphylococcus aureus, Salmonella schotmuehleri, Klebsiella pneumoniae, Diplococcus pneumaniae and Streptococcus pyogenes.

Examples of compounds of general formula III are 7-[1,2,5-diamino-2-difluoromethylvaleryl) amino] -3-acetyloxymethyl-8-oxo-5-thia-1-azabicyclo (4.2,0) oct-2-ene -2-carboxylic acid.

Preparation of the compounds of general formula III is described below.

As pharmacologically useful agents, compounds of general formula I and II can be administered in various ways to patients to be treated to achieve the desired effect. The compounds can be administered alone or in combination with each other.

The compounds may also be administered in the form of pharmaceutical preparations. The compounds may be administered orally, parenterally, for example intravenously, intraperitoneally or subcutaneously or topically.

The amount of compound administered varies over a wide range and can be any effective amount. Depending on the patient to be treated, the condition to be treated, the method of administration, the effective amount of the compound to be administered will vary from about 0.1 mg / kg to 500 mg / kg of body weight per unit dose and preferably 10-100 mg. / kg body weight per unit dose. A typical unit dosage form may be, for example, a tablet containing from 10 to 300 mg of a compound of formula I or II, which is administered to the patient 1-4 times daily to achieve the desired effect.

"Patient" here denotes a warm-blooded animal such as mammals, for example cats, dogs, rats, mice, guinea pigs, horses, cattle and sheep, and humans.

The solid unit dosage forms can be of the conventional type, thus solid forms may be a capsule which may be of the normal gelatin type containing a novel compound of the present invention and a carrier, for example a lubricant and inert filler such as lactose, sucrose and cornstarch. In other embodiments, the novel compounds can be tableted with conventional tablet bases, such as lactose, sucrose and corn starch in combination with such binders as acacia gum, corn starch and gelatin, disintegrants such as corn starch and potato starch and alginic acid and lubricating agents such as stearic acid and magnesium.

For parenteral administration, the compounds may be administered in injectable doses of a solution or suspension of the compound in a physiologically acceptable diluent with a pharmaceutical carrier, which may be of sterile liquid such as water or an oil with or without the addition of surfactants or other pharmacologically acceptable additives. Examples of oils that can be used in these preparations are oils of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil and mineral oil. Generally, water, saline, aqueous solutions of dextrose and similar sugar solutions, ethanol and glycols such as propylene glycol or polyethylene glycol are preferred as liquid carriers, especially for injectable solutions.

The compounds may be administered in the form of a depot injection or implant, which may be formulated to allow prolonged release of the active ingredient. The active ingredient can be applied to pellets or small cylinders and implanted subcutaneously or intramuscularly as depot injections or implants. The implants can be inert materials such as biodegradable polymers or synthetic silicones, for example Silactis, silicone rubber manufactured by Dow Corning Corporation.

The new compounds of the general formula II, in which Z represents H2N (CH2) 3 and R1 represents hydroxy, are prepared by treating an ester derivative of ornithine, in which the amino group or groups 1307691-6 are suitably protected, may react with a suitable halo gene methyl haloalkylating reagent in an aprotic solvent such as dimethylsulfoxide, dimethylformamide, dimethylacetamide, benzene, toluene, ethers such as tetrahydrofuran, diethyl ether or dioxane at a temperature between -120 to 120 ° C, preferably 25-SOOC, for up to 48 hours , followed by acidic or basic hydrolysis and represented by the following reaction sequence: I "1 _ _ (_) X1 f fl COOR6 'z1-c - cooR6: N = cR II | I 7 strong base g IN = C-R7 I ----- 1 Rs 'i I |. 8 I Compound 1 _ _' "'' alkylation- CHF2> reagent 21-c-cooR6; çHF2 NH I 2 diluted water- Z1-Q-COOR6_ solution IN = c- R7 Compound 3 | acid / hydrazine R8 H2O (acid / base) QHFZ Compound 2 z1-c-coon NH2 Formula IV In the above reaction scheme, R6 represents a lower alkyl group, for example methyl, ethyl, isopropyl, n-propyl or n-butyl; R 7 represents hydrogen, phenyl, a straight or branched alkyl group having 1 to 8 carbon atoms, methoxy or ethoxy; R 8 represents phenyl or a straight or branched alkyl group having 1-8 carbon atoms, or R 7 and R 8 together form an alkylene group having 5-7 carbon atoms, i.e. -CH 2 - (CH 2) m -CH 2 -, where m represents 3, 4 or 5. Examples of straight or branched alkyl groups having 1-8 carbon atoms, which R7 and R8 may represent, are, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, neopentyl or triethyl. 7807691-6 '8 methyl; where z represents R (cn) -, where R represents -N = c-R, 1. 10 2 3 10 a, 7. RB where R 7 and R 8 have the same meaning as stated above.

When in compound 1 Z 1 represents R 10 (CH 3) 3 -, where R 10 represents -N = ç-R 7, R 7 and R 8 are the same.

R8- Removal of the protecting group from the amine, mercapto and carboxyl functions can be done in one step by treating the compound Z with an aqueous solution of acid, for example hydrochloric acid or toluenesulfonic acid having a temperature of from about 0 to 160 ° C below 4 to 24 hours, whereby a compound of the general formula IV is obtained. It is preferred to first remove the protecting groups from the amine functions or the function of compound 2 when said functions are protected as Schiffs bases by treating the compound 2 with dilute aqueous solution of an acid, for example hydrochloric acid or with hydrazine or phenylhydrazine in solvent. such as lower alcohols, for example methanol or ethanol, ethers, chlorinated hydrocarbons, benzene and water. Removal of the protecting groups in the carboxyl functions and the amine groups when the amine groups are protected other than by Schiffs base is achieved by treating the compound 3 with a concentrated aqueous solution of an acid, for example hydrobromic acid and a temperature from 0 to 160 DEG C. in aqueous solutions of bases, for example ammonium hydroxide.

Compounds of the general formula I, in which Z represents Y-guanidinopropyl are prepared from corresponding suitably protected derivatives, in which Z represents H2N (CH2) 3 -? HF2 H2N (CH2) 3 -? - COOH Formula V NH2 by treatment with an alkylisothiouronium salt, for example ethyl isothiouronium hydrobromide by methods which are generally known, for example Organic Organic Synthesis III, p. 440 (1955). The reaction takes place in the presence of a base, for example an aqueous solution of 7807691-6 in sodium hydroxide or potassium hydroxide at a pH of about 8-12 at a temperature of from about 0 to 10,000 for 6 hours to 8 days, after which the reaction mixture is neutralized with concentrated mineral acid, such as hydrochloric acid, and the product is isolated. The α-amino group may be protected with, for example, a benzyloxycarbonyl group.

The amine protecting group is then removed by acid hydrolysis, for example with hydrochloric acid; The compounds of general formula I, wherein R 1 is a straight or branched alkoxy group having 1 to 8 carbon atoms, are prepared by conversion of the corresponding compound, wherein R 1 represents hydroxy to an acid halide by, for example, treatment with thionyl chloride, followed by an alcoholysis with an alcohol of the formula R 17 OH, wherein R 17 is a straight or branched alkyl group having 1-8 carbon atoms in a manner known per se. Alternatively, compounds of general formula I, wherein R 1 is a straight or branched alkoxy group having 1-8 'carbon atoms may be prepared with the corresponding derivatives, wherein R 1 represents hydroxy by treating said derivative with an alcohol of the formula R 17 OH as defined above saturated with hydrogen chloride for 30 minutes to 12 hours at a temperature from about 25 ° C to the boiling point of the alcohol.

The individual optical isomers of the compounds of formula I, where Z represents Y-guanidinepropyl, are obtained in the manner described here for the racemates starting from the dissolved orgitin analogue.

The compounds of general formula III are prepared by coupling a 7-aminocephalosporanic acid or a derivative thereof of the formula N * / CH X Formula IX COOM where M represents hydrogen or a negative mixture and X represents hydrogen or acetoxy, with an acid of formula 7807691-6, 10 CHF '-2 ZFC-COOH Ä NH Formula I 2 or a functional derivative thereof, such as the acid chloride or an acid anhydride in the presence of a dehydrating agent such as dicyclohexylcarbodiimide, when the free acid is used and the amino group is protected with a suitable blocking group, for example tert-butoxycarbonyl followed by acid hydrolysis to remove the protecting amino group.

The coupling reaction is generally carried out in a solvent such as ethyl acetate, dioxane, chloroform or tetrahydrofuran in the presence of a base such as alkali bicarbonate. The temperature of the reaction can vary from -100 to 100 ° C, and the reaction time can vary from about an hour to 10 hours. The cephalosporin products are isolated in a conventional manner. The compounds of formula I are prepared by methods described above and the compounds of formula IX are commercially available.

The following examples illustrate the use of a compound of general formula I, wherein R 1 represents hydroxy as a chemical intermediate in the preparation of cephalosporins of formula III.

Example 1 1 7- [X2,5-Diamino-2-difluoromethylvaleryl) amino] -3-acetyloxy-methyl-8-oxo-5-thia-1-azabicyclo [1,2] octo-2-ene-2-carboxylic acid A mixture of 1 g of 3-acetyloxy-7-amino-8-oxo-5-thia-1-azabicyclo [α] 2,2-octo-2-ene-2-carboxylic acid and 1 g of 2,5-diamino-2-difluoro- methylvaleryl acid and 1 g of 2,5-diamino-2-difluoromethylvaleric acid chloride, where the free amino group is protected with tert-butoxycarbonyl in 50 ml of ethyl acetate is refluxed for 2 hours, after which the solvent is removed to leave a residue which is treated under slightly acidic conditions and chromatographed on silica gel using benzene-acetone as eluent to give 7- [X2,5-diamino-2-difluoromethylvaleryl) amino] -3-acetyloxymethyl-8--oxo-5-thia- 1-Azabicyclo [1,2,2-fokt-2-ene-2-carboxylic acid. Example 2 Composition for hard gelatin capsules as follows: (a) u, 6Ediamino-d-difluoromethylvaleric acid l 20 mg (b) talc 5 mg (c) lactose _ 90 mg The preparation is prepared by the dry powder of (a) and (b) is allowed to pass through a fine sieve to mix well. The powder is then filled into hard gelatin capsules in an amount of 115 mg per capsule.

Example 3 Tablet composition: (a) d-amino-d-fluoromethyl-6-guanidinovaleric acid 20 mg (b) starch 43 mg (c) lactose 45 mg (d) magnesium stearate 2 mg The granulation was obtained by mixing the lactose with the compound (a) and a portion of the starch and granules with starch paste which were dried, sieved and mixed with magnesium stearate. The mixture was beaten into tablets weighing 110 mg each.

Example 4 Composition for an injectable suspension based on 1 ml ampoule for an intramuscular injection.

Weight percent (a) α-amino-d-difluoromethyl-6-methylthiobutyric acid 1.0 (b) polyvinylpyrrolidone 0.5 (c) lecithin 0.25 (d) water for injection 100.0 The materials (a) - (d) were mixed , homogenized and filled into 1 ml ampoules, which were sealed and autoclaved for 20 minutes at 121 ° C. Each vial contained 10 mg per ml of new compound (a).

Example 5 2-Difluoromethyl-2,5-diaminopentanoic acid In a nitrogen atmosphere, a solution (500 ml) of ZM butyllithium in hexane was added to a stirred solution of 143.1 ml of diisopropylamine in 1.5 l of tetrahydrofuran at 7878 ° C, after to which 261 g (0.81 mol) of ornithine dibenzaldimine ester in 1.5 liters of tetrahydrofuran were added. After completion of the addition, the reaction temperature was raised to 40 ° C and kept between 40 and 50 ° C for 3 hours during which time chlorodifluoromethane was bubbled through the mixture with stirring. The reaction mixture was then treated with a saturated solution of sodium chloride. The organic material was extracted with ether and the ether extract was washed several times with sodium chloride solution and dried over magnesium sulphate and evaporated to give a viscous oil. The oil was stirred with 1N HCl (1.5 L) for 3 hours, after which the mixture was extracted several times with chloroform and the aqueous solution was evaporated to dryness. The oily residue was refluxed with 12N hydrochloric acid (1.5 L) for 16 hours, after which the cooled solution was clarified by chloroform extraction before being concentrated, decolorized (activated carbon) and further concentrated to about 750 ml. The pH of the solution was adjusted to 3.5 by the addition of triethylamine, the solution was treated again with activated carbon before concentration to 500 ml and diluted with 7-8 liters of acetone. The precipitated product was filtered off and washed with ethanol. The crude product was recrystallized by dissolving in about 150 ml of hot water and treating with a solution of hot ethanol (450 ml). Upon cooling, 2-difluoromethyl-2,5-diaminopentanoic acid hydrochloride monohydrate crystallized separately as 71 g (37%), mp 183 ° C.

Claims (4)

10 15 20 25 30 35 H 7807691 ”6 Patent claims 1. l. A compound of the formula CHF 1 2 Z -? COR NH2 1 where Z represents Y-guanidinopropyl and R1 represents hydroxy or where Z represents H2N (CH2) 3 and R1 represents hydroxy or a straight or branched alkoxy group having 1 to 8 carbon atoms and pharmacologically acceptable salts and individual optical isomers thereof. A compound according to claim 1, characterized in that it is 2-difluoromethyl-2,5-diaminovaleric acid or a pharmacologically acceptable salt thereof. A compound according to claim 1, characterized in that it is 2-difluoromethyl-2-amino-5-guanidinopentanoic acid or a pharmacologically acceptable salt thereof. A process for the preparation of a compound of the formula CHF | 2 Z - C - COR1 h; 2 wherein Z represents Y-guanidinopropyl and R 1 represents hydroxy or where Z represents H 2 N (CH 2) 3 and R 1 represents hydroxy or a straight or branched alkoxy group having 1 to 8 carbon atoms and pharmacologically acceptable salts and individual optical isomers thereof. according to claim 1, characterized in that step (a) when Z represents H 2 N (CH 2) 3 - and R 1 represents hydroxy. that an ester derivative of ornithine. where the amino groups are suitably protected. is allowed to react with a strong base to form a carbonate intermediate. which is reacted with a suitable halomethyl-haloalkylating reagent in an aprotic solvent and at a temperature of about -120 to 120 ° C for 1/2 to 48 hours followed by acidic or basic hydrolysis; (b) when Z represents Y-guanidinopropyl. the corresponding suitably protected derivative wherein Z represents H 2 N (CH 2) 3 is treated with an alkyl isothiouronium salt in the presence of a base at a pH of from about 8 to 12 and a temperature of from about 0 to 100 ° C for 6 hours to 8 days followed by neutralization with concentrated hydrochloric acid: (c) when R is a straight or branched alkoxy group having 1 to 8 carbon atoms, conversion of the corresponding derivative where R1 represents hydroxy to the acid halide followed by alcoholysis with an alcohol of the formula R wherein R 17 17 group having 1-8 carbon atoms: is a straight or branched alkyl- (d) when a pharmaceutically acceptable salt is desired the compound thus obtained is reacted with a suitable pharmacologically acceptable acid or has.