FI67378B - FOR THERAPEUTIC TREATMENT OF THERAPEUTIC THERAPEUTIC THIAZOLIDES - Google Patents

Description

RSF ^ l M «« ADVERTISEMENT 67 37 8 ^ rjk lBJ * "> utlAggningsskkift ° '° c (45) t': .. i uy ^ nn ·: lly 'i lv ^ 5 sS £ b2V'“ c 07 D 283 / 00 // C 07 ο 277/06, v (51) K ** - / ««. A. 279/06, 279/12, 265/30 FINLAND — FINLAND pi) ^ or ^ -p «« wew < 7736 ^ 0 (12) 0,., 2.77 V 7 (23) AHcuplM — GIWftMtadag 01.12.77 (41) Tullut lulUMksI - BIlvH aff «ntN | 0 * +.

Patent and Rector Rated ...,,,. „,. . . . (44) Date of decision on the date of entry into force of this Regulation. - n,

Patent and Register AmBkan utlagd odi uti ^ kitfMn pubiiearad 30.11 .oh (32) (33) (31) PjnMsejr utuoikaui —8 * fird prtorltet 03.12.76 23.09.77 USA (US) 71 + 7281, 836107 (71) ER Squibb & Sons, Inc., Lawrencevi1le-Princeton Road, Princeton,

New Jersey 085 * 10, USA (US) (72) Miguel Angel Ondetti, Princeton, New Jersey, USA (US) (7 * +) Oy Kolster Ab (54) Method for the preparation of new therapeutically useful thiazolidine and thiazane derivatives - Förfarande för framställ -ing a therapeutic alternative to thiazolidine- and thiazander derivatives

The invention relates to a process for the preparation of new therapeutically active thiazolidine and thiazanecarboxylic acids of the formula I and their basic salts, X, / \ j * 3 »Vpm <pVn

R4 - S - (CH2) - CH - CO - N ---- CH - CO - OH I

wherein R 1 and R 2 represent hydrogen or lower alkyl; is hydrogen or lower alkyl; is hydrogen, lower alkanoyl or benzoyl; X is O, S, SO or SO 2> wherein m is 2 and n is 1 when X is O; m is 1 or 2, n is 0, 1 or 2 and 67378 m + n is 2 or 3; p is 0 or 1.

The compounds of the formula I are characterized by an unsubstituted or lower alkyl-substituted 5- or 6-membered heterocyclic carboxylic acid having one nitrogen atom and one sulfur or oxygen atom in the ring, the remaining ring members being carbon atoms, with thiazolidines being preferred. -sanic and morpholinecarboxylic acids. As shown, in addition to nitrogen, the ring contains a heteroatom which is oxygen or sulfur or sulfur,

O

II

which may be oxidized to sulfinyl (-S-) or sulfonyl (O- (S-)). The side chain attached to the nitrogen of the heterocyclic ring is an unsubstituted or substituted mercaptoalkanoyl group.

The group of lower alkyls represented by one of the alternatives includes straight-chain or branched hydrocarbon radicals from methyl to heptyl, e.g. methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl and the like. Lower alkylene groups are similar, including 1 to 7 carbon atoms. Likewise, lower alkoxy groups are similar groups attached to an acid, e.g. methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, t-butoxy and the like, C 1-4 groups, especially all types of C 1-4 and C 1-4 groups take precedence. Lower alkanoyl groups are acyl radicals of lower fatty acids (up to 7 carbons), e.g., acetyl, propionyl, butyryl, and the like, of which acetyl is the primary group.

Preferred are those compounds of formula I wherein R is hydroxy; R 1 and R 2 are in particular hydrogen, methyl or ethyl, in particular hydrogen; especially hydrogen, methyl or ethyl; I <4 is especially hydrogen, acetyl or benzoyl; X is sulfur or oxygen, especially sulfur; m is 1 or 2; n is 1; and p is 0 or 1, especially 1.

The products of formula I are prepared according to the invention in such a way that a) a compound of formula II is (X-CH) "(CH-R 1)

2 | m | 1 n II

UN ---------- CH - COOH

acylated together, in a step by reacting it with a compound of formula III or a reactive equivalent thereof, 3 67378 R-, I 3

R. - S - (CHO) - CH - COOH III

4 2 p or b) the compound of formula II is gradually acylated by first adding a part of the acylating agent III to an acid of formula II, e.g. by reacting this acid with a haloacyl halide of formula (IIIa) R3 is - (CH-) -CH- CO-hal (Ula)

2 P

wherein the shark represents halogen, preferably chlorine or bromine, e.g. 3-bromopropanoyl chloride, to give the product of the following formula (IIIb): (x-R- (R0-CH) (CH-R.) (IIIb) 13 2 | m | in

hai- (CH-) -CH-CO- N --- CH-CO-OH

2 p which is then reacted with thiol-R 1 -SH to give the desired product of formula I, optionally followed by hydrolysis of the compound of formula I thus obtained, in which R 1 is other than hydrogen, to a compound of formula I in which R is hydrogen, and / or optionally oxidizing a compound of formula I wherein X is S to a compound of formula I wherein X is SO or SO 2, or a compound of formula I wherein X is SO is oxidizing to a compound of formula I wherein X is SO2, and the compound of formula I thus obtained is optionally converted into a basic salt.

The acylation reaction uses known methods to activate acid III prior to reaction with acid II, including mixed anhydride, symmetrical anhydride, acid chloride, active ester Woodward's reagent K, N, N'-carbonyl-bisimidazole, EEDQ (N -methoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) or the like.

4 67378

The products of formula I have at least one or may always have four asymmetric carbon atoms. These carbon atoms are marked with an asterisk in Formula I. Accordingly, the compounds exist in diastereoisomeric forms or as racemic mixtures thereof. They are all within the scope of the invention. In the synthesis described above, a racemate or one of the enantiomers can be used as a starting material.

After the racemic starting material has been used in the synthesis, the stereoisomers obtained as a product can be separated by conventional chromatographic methods or by fractional crystallization methods. In general, the L-isomer with respect to the amino acid carbon is the preferred isomeric form.

The compounds of the invention form basic salts with various inorganic and organic bases, which are also within the scope of the invention. Such salts include ammonium salts, alkali metal salts such as sodium and potassium salts (which are preferred), alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases, e.g. dicyclohexylamine salts, benzathine, N-methyl-D- glucamine, hydrabamine salts, salts of amino acids such as argonine and lysine, and the like, bi-toxic, physiologically acceptable salts are preferred, although other salts are also useful, e.g., in isolating and purifying the product, as in the case of the dicyclohexylamine salt.

The salts are formed by a conventional method in which the free acid form of the product reacts with one or more base equivalents containing the desired cation in a solvent or medium in which the salt is insoluble and filterable, or in water and water is removed by lyophilization. By neutralizing the salt with an insoluble acid such as a cation exchange resin in the form of a hydrogen ion, a polystyrene sulfonic acid resin - Dowex 50 (Mikes, Laboratory Handbook of Chromatography Methods, Van Nostrand, 1961) page 256./ or an aqueous solution of the acid, and extract with an organic solvent, and extract with an organic solvent. ., a free acid can be obtained, and if desired, another salt is formed.

Further details of the experiment can be found in the examples, which are preferred embodiments of the patent and also serve as models for preparing other members of the group.

67378

The compounds of the invention inhibit the conversion of the decapeptide, angiotensin I, to angiotensin II and are therefore useful in reducing or alleviating angiotensin-related hypertension. When the enzyme renin acts on angiotensinogen, plasma psendoglobulin, angiotensin I is formed. Angiotensin I is converted by angiotensin "converting" enzyme (ACE) to angiotensin II. The latter is an active antihypertensive agent that causes hypertension in various mammalian species, e.g., rats, dogs, etc. The compounds of the invention are coupled to a series of reactions or preventing its formation.

Angiotensin "converting" enzyme inhibition compounds of formula I can be measured in vitro using an angiotensin "converting" enzyme isolated from rabbit lung using the procedure described by Cushman and Cheung ^ Biochem. Pharmacol., 20, 1637 (1971) J and by determining soft muscle / E. O'Keefe, et al., Federation Proc. 31, 511 (1972), 7, in which these compounds have been shown to be potent inhibitors of angiotensin I-induced contraction and bradykinin-induced contraction enhancers.

When a composition comprising a single compound of formula I or a combination of compounds of formula I or a physiologically acceptable salt thereof is administered to a species of hypertensive mammal, it relieves or reduces the elevated pressure due to angiotensin. A single dose, or preferably a daily dose divided into two or four portions, with a daily dose of 5 to 1000 mg, preferably 10 to 500 mg per kilogram, suitably lowers blood pressure. Model experiments performed on animals as described by S.L. Engel, T.R. Schaeffer, M.H. Waugh and B. Rubin,

Proc. Soc. Exp. Biol. Med. 143, 483 (1973), are useful guidelines.

The agent is preferably administered orally, but parenteral routes such as subcutaneous, intramuscular, intravenous or intraperitoneal administration may also be used.

The compounds of the invention may be used to lower blood pressure by formulating them, e.g., as tablets, capsules or elixirs for oral administration or as sterile solutions or suspensions for parenteral administration. About 10 to 500 mg of a compound of formula I or a mixture of a compound of formula I or a physiologically acceptable salt thereof is combined with a physiologically acceptable excipient, carrier binder, preservative, stabilizer, spice, etc. in unit dosage form as required by acceptable pharmaceutical techniques. The amount of active ingredient in these compositions or preparations is such that a suitable dosage will be obtained within the stated range.

The following examples illustrate the invention and are preferred embodiments of the patent. All temperatures are in degrees Celsius.

Example 1 3- (3-Benzoylthiopropanoyl) -4-L-thiazolidinecarboxylic acid To an ice-cooled solution of L-4-thiazolidinecarboxylic acid (6.6 g) in 1N NaOH (50 ml) is added 2N sodium hydroxide (25 ml). ) and 3-bromopropionyl chloride (8.5 g) in that order, with vigorous stirring. After 3 hours, a suspension of thiobenzoic acid (7.5 g) and potassium carbonate (4.8 g) in water (50 ml) is added. The reaction mixture is stirred at room temperature overnight and the mixture is filtered. The filtrate is acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The organic layer is dried and concentrated to dryness. The residue is purified by silica gel chromatography (benzene: acetic acid, 7: 1) and the purified material is crystallized from ethyl acetate-ether-hexane to give 3- (3-benzoyl-thiopropanoyl) -4-L-thiazolidinecarboxylic acid, m.p. 105-106 °.

The above product is dissolved in water and an equivalent amount of sodium hydroxide solution is added. The solution is lyophilized to give the sodium salt.

Example 2 3- (3-Mercaptopronoyl) -L-4-thiazolidinecarboxylic acid 3- (3-Benzoylthiopropanoyl) -L-4-thiazolidinecarboxylic acid (6.7 g) is dissolved under water under argon (15 ml) and concentrated. to a mixture of ammonia (7.5 ml). After storage at room temperature for one hour, the reaction mixture is diluted with water (20 ml) and filtered. The filtrate is extracted with ethyl acetate, acidified with concentrated hydrochloric acid and re-extracted with ethyl acetate. The second ethyl acetate extract is dried and concentrated to dryness. The residue, 3- (3-mercaptopropanoyl) -L-4-thiazolidinecarboxylic acid is crystallized from ethyl acetate, m.p. 110-112 °.

Example 3 a) 3-Acetylthio-2-methylpropionic acid (starting material)

A mixture of thioacetic acid (50 g) and methacrylic acid (40.7 g) is heated on a steam bath for one hour and then stored at room temperature for eleven hours. The reaction mixture is distilled in vacuo and the fraction b.p. is 128.5-131 °, 2.6 mm.

3-Acetylthio-2-methylpropionic acid can also be isolated by allowing the reaction mixture to crystallize after dilution with hexane, m.p. 40-42 °.

b) 3- (3-Acetylthio-2-methylpropanoyl) -2-thiazolidinecarboxylic acid methyl ester 2-Thiazolidinecarboxylic acid methyl ester (CA 53, 12,281d) (4.4 g) and 3-hydroxybenzotriazole (4.0 g) are dissolved dichloromethane (40 ml) and the solution is stirred and cooled in an ice bath. Dicyclohexylcarbodiimide (6.2 g) is added, dissolved in dichloromethane (15 ml) and immediately followed by a solution of 3-acetyl-2-methylpropionic acid (4.9 g) in dichloromethane (5 ml). After stirring in an ice bath for 15 minutes and at room temperature for 16 hours, the precipitate is removed by filtration and the filtrate is washed until neutral. The organic layer is dried and concentrated to dryness in vacuo to give 3- (3-acetylthio-2-methylpropanoyl) -2-thiazolidinecarboxylic acid methyl ester.

c) 3- (3-Mercapto-2-methyl-propanoyl) -2-thiazolidinecarboxylic acid 3- (3-acetylthio-2-methyl-propanoyl) -2-thiazolidinecarboxylic acid methyl ester (2.9 g) is dissolved in methanol (30 ml) and N sodium hydroxide (30 ml) is added. The reaction mixture is stirred at room temperature, a portion is removed every hour and checked by paper electrophoresis for methyl ester hydrolysis ratio. When this hydrolysis is complete (about three hours), the reaction mixture is neutralized, concentrated in vacuo to remove methanol, acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The organic layer is dried and concentrated to dryness to give 3- (3-mercapto-2-methylpropanoyl) -2-thiazolidinecarboxylic acid, m.p. 96-101 ° C.

Calculated for C 9 H 11 NO 3: C 40.83, H 5.57, N 5.95, S 27.25 Found: C 40.45, H 5.38, N 5.73, S 26.97 SH- titration 99.4%.

Example 4 3- (3-Mercaptopropanoyl) -1,3-thiazane-4-carboxylic acid

By substituting 1,3-thiazane-4-carboxylic acid for 4-L-thiazolidinecarboxylic acid in the process of Example 1.

Biol. Chem. , 607 (1957) and using the product in the method of Example 2, 3- (3-benzoylthiopropanoyl) -1,3-thiazane-4-carboxylic acid and 3- (3-mercaptopropanoyl) -1,3-thiazane-4- carboxylic acid.

The final product (3-mercapto compound) is obtained as a pale yellow, viscous syrup with an Rf value of 0.32 (silica gel-benzene / acetic acid (7: 1)) and the following elemental analysis: Calculated for C8H13No3S2: C 40, 83, H 5.57, N 5.95, S 27.25 Found: C 40.96, H 5.54, N 6.02, S 26.94.

Example 5 3- (3-Mercapto-2-methylpropanoyl) -L-4-thiazolidinecarboxylic acid 3-Acetylthio-2-methylpropanoic acid chloride (5.4 g, prepared from 3-acetylthio-2-methylpropanoic acid and thionyl chloride, b.p. 80 ° C). ) and 2-n sodium hydroxide (15 ml) are added to a solution of L-4-thiazolidinecarboxylic acid (Z. Naturforschg., 17b (1962) 765) (5.2 g) in 1-n sodium hydroxide (30 ml), which is chilled in an ice / water bath. The mixture is stirred for 3 hours at room temperature, then extracted with ether, the aqueous phase is acidified and extracted with ethyl acetate. The organic phase is dried over magnesium sulfate and concentrated to dryness in vacuo to give 3- (3-acetylthio-2-methylpropanoyl) -L-4-thiazolidinecarboxylic acid.

9 67378 3- (3-Acetylthio-2-methylpropanoyl) -L-4-thiazolidinecarboxylic acid (1 g) is dissolved in a mixture of water (3 ml) and concentrated ammonia (3 ml) under argon. The mixture is stirred at room temperature for 30 minutes and acidified with concentrated hydrochloric acid. The organic layer is concentrated to dryness in vacuo to give 3- (3-acetylthio-2-methylpropanoyl) -L-4-thiazolidinecarboxylic acid (dicyclohexylammonium salt crystallized from acetonitrile, mp 172-186 ° C (m.p. 130 ° C)) and 3-. (3-mercapto-2-methylpropanoyl) -L-4-thiazolidinecarboxylic acid (ethyl acetate / dicyclohexylammonium salt crystallized from hexane, mp 180-188 ° C (sintering 170 ° C)).

Example 6 4- (3-Acetylthiopropanoyl) -L1,4-thiazane-5-carboxylic acid L-4-thiomorpholine-2-carboxylic acid hydrochloride (6.6 g, 0.036 m) is dissolved in 150 ml of dimethylacetamide and 3- acetylthiopropanoyl chloride (5.97 g, 0.036 m). The temperature is raised to 28 °. To this solution is added N-methylmorpholine (10.9 g, 0.108 m). The temperature is raised to 42 °, whereupon a white precipitate forms immediately. The mixture is heated on a steam bath for one hour and allowed to stand at room temperature overnight. The solid is removed by filtration to give 9.7 g of 4- (3-acetylthiopropanoyl) -L-1,4-thiazane-5-carboxylic acid, m.p. 202-204 °. The solvent is removed to give a viscous residue which is triturated with water and 20% hydrochloric acid. The precipitated oil is extracted with 3 x 150 ml of ethyl acetate and the extracts are dried over magnesium sulfate. The solvent is removed and the viscous residue (7.5 g) crystallizes on standing. After recrystallization from acetone-hexane, the product has a constant melting point of 122-125 °.

Example 7 4- (3-Mercaptopropanoyl) -L-1,4-thiazane-5-carboxylic acid

Aqueous ammonia (13 mL of concentrated ammonium hydroxide in 30 mL of water) is stirred under nitrogen for 15 minutes and solid 4- (3-acetylthiopropanoyl) -L-1,4-thiazane-5-carboxylic acid (6.8 g, 0.024 m) is added. A clear solution suddenly forms at 5-10 °. The solution is stirred at room temperature under a nitrogen atmosphere for one hour. The solution is extracted with 100 ml of ethyl acetate 67378 and the aqueous layer is strongly acidified with 20% hydrochloric acid. The precipitated oil is extracted with 3 x 150 ml of ethyl acetate. The extracts are combined and dried over magnesium sulphate, then the solvent is removed to give 5.6 g of a crystalline material which appears to contain considerable starting material. The substance obtained (5.5 g) is rehydrolyzed for a further two hours with a solution of, as mentioned above, 12 ml of concentrated ammonium hydroxide in 25 ml of water. The solution is acidified and the precipitated oil is extracted with 3 x 150 ml of ethyl acetate. The extracts are combined and dried over magnesium sulphate, then the solvent is removed to give 2.7 g (48%) of 4- (3-mercaptopropanoyl) -L-1,4-thiazane-5-carboxylic acid as a viscous mass, after drying overnight. above room temperature and 1 mm.

Analysis calculated for C 10 H 10 NO - Sn: N, 5.95, C, 40.82; H, 5.56, 0.1 JOL · S, 27.25, SH, 100% Found: N, 6.13, C, 40.85, H, 5.46, S, 27.38, SH, 96% .

Example 8 3- (3-Mercapto-2-D-methyl-1-oxopropyl) -L-4-thiazolidinecarboxylic acid 1-oxide a) 3- (3-Acetylthio-2-D-methyl-1-oxopropyl ) -L-4-thiazolidinecarboxylic acid 1-oxide 2.76 g of 3- (3-acetylthio-2-D-methyl-1-oxopropyl) -L-4-thiazolidinecarboxylic acid are added to a mixture of 20 ml of methyl -leneyl chloride and 1.84 ml of formic acid, stirring at room temperature. To this mixture is added 1.04 ml of 30% hydrogen peroxide. After 3.5 hours, the mixture is concentrated to dryness in vacuo, treated with ethyl acetate, filtered and concentrated to dryness in vacuo to give 2.5 g of product.

b) 3- (3-Mercapto-2-D-methyl-1-oxopropyl) -L-4-thiazolidinicarboxylic acid 1-oxide

The material from step a) is treated for 20 minutes with a cold solution of water (20 ml) and concentrated ammonium hydroxide (10 ml), acidified with AG50WX2 resin and chromatographed on AG50WX2 resin to give 1 g of product as a hygroscopic lyophilisate with optical rotation. / oc_7D is -183.1 (c = 1.42, in methanol).

Analysis for C 18 H 28 N 2 O 2 S 2.1 / 2 1 ^ 0

Calculated: C 36.95, H 5.43, N 5.39, S 24.66 Found: C 36.94, H 5.45, N 5.45, S 24.82.

6737 8

Example 9 3- (3-Mercapto-1-oxopropyl) -2-methyl-L-4-thiazolidinecarboxylic acid 3- (Acetylthio) propionyl chloride (9.0 g, 0.07 m) is slowly added to a solution of 2- methyl L-4-thiazolidinecarboxylic acid (8.0 g, 0.0544 m) and sodium carbonate (2.9 g) in 70 ml of distilled water while maintaining the pH at 9 at 6 ° C by adding 24% sodium carbonate solution. The mixture is stirred for 30 minutes in an ice bath, then 70 ml of water are added to give a clear solution. Stirring is continued at room temperature for one hour, after which the mixture is extracted twice with ethyl acetate and cooled to 6-7 ° C, acidified to pH 2.5 with concentrated hydrochloric acid. The reaction mixture is then saturated with sodium chloride and extracted twice with ethyl acetate. The ethyl acetate extracts are dried over magnesium sulphate and evaporated to dryness to give 13.32 g of 3- (3-acetylthio) -1-oxopropyl) -2-methyl-L-4-thiazolidinecarboxylic acid, which is then converted into the dicyclohexylamine salt, crystallized from isopropanol and further converted into isopropanol. melting at 119-122 ° C (crystallized from ethyl acetate / hexane).

The product obtained above is added to a mixture of 12 ml of water and 8 ml of ammonium hydroxide in an ice bath, and then the mixture is stirred for one hour at room temperature, then 40 ml of water are added to the mixture, and the mixture is extracted twice with ethyl acetate. The aqueous layer is then cooled, acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The ethyl acetate layer is dried over magnesium sulfate and concentrated to dryness in vacuo to give 2.3 g of an oil which is then stirred in hexane for 72 hours to give 1.75 g of 3- (3-mercapto-1-oxopropyl) -2-methyl- L-4-thiazolidinecarboxylic acid as a colorless material, m.p. 69-95 ° C.

Analysis for C 9 H 8 S 2 NO 9

Calculated: C 40.83, H 5.57, S 27.25, N 5.95 Found: C 40.96, H 5.60, S 27.39, N 5.89.

67378 12

Examples 10 and 11 D, L- [3- (acetylthio) -1-oxopropyl] -7-2-thiazolidinecarboxylic acid and dicyclohexylamine salt (1: 1) a) DL-thiazolidine-2-carboxylic acid

A mixture of 19.0 g (0.167 mol) of cysteamine hydrochloride and 14.9 g (0.201 mol) of glyoxylic acid hydrate is dissolved in 15% water. 50 ml of anhydrous ethanol are added to the mixture, after which the reaction mixture is allowed to stand at room temperature for 45 minutes and then treated with 13.8 g (0.168 mol) of sodium acetate. The reaction mixture is then allowed to stand at room temperature overnight, and then cooled for one hour, and then filtered using chilled 95% ethanol for washing. The product is dried and the crude product (16.4 g) had a melting point of 178-180 ° C. (Dec.). The analysis showed that the crude product contained about 0.93 molar equivalents of sodium chloride.

Analysis for C 4 H 7 NC> 2.0.93 NaCl

Calculated: C 25.62, H 3.76, N 7.47, S 17.10, Na 11.40, Cl 17.58 Found: C 25.79, H 3.99, N 7.32, S 17 , 36, Na 11.59, Cl 18.21.

b) DL-3- [13- (acetylthio) -1-oxopropyl] -2-thiazolidinecarboxylic acid dicyclohexylamine salt

To a mixture of 56 ml of water and 63 ml of tetrahydrofuran cooled in an ice / water bath are added 7 g (37.3 mmol) of thiazolidine-2-carboxylic acid (.0.93 NaCl) and 9.1 g (112 mmol) of ) sodium bicarbonate. With vigorous stirring and cooling, 5.0 ml (37.4 mmol) of 3-acetylthiopropionyl chloride are added dropwise. During the addition, the pH is maintained close to 7 by the addition of solid sodium bicarbonate. After 45 minutes, the pH ceases to change, at which point the reaction is terminated by the addition of 1N HCl to pH 1, after which the reaction mixture is extracted with ethyl acetate. The organic layer is washed with water and brine, dried over sodium sulfate and concentrated in vacuo to give 10.0 g of crude product. The product was chromatographed using 850 g of Baker silica gel and CHCl: MeOH: HOAc (20: 2: 1) as eluent to give 9.2 g of crude product which showed small amounts of impurities and a small amount of solvent residues by thin layer chromatography.

The chromatographed free acid (9.3 g) is dissolved in 6 ml of 67378 ethyl acetate and this solution is added to 300 ml of ether. About 200 ml of ethyl acetate are added to give a clear solution from which 13.3 g (89%) of analytically pure product precipitates within two hours at room temperature, m.p. 195-196 ° C and neutralization equivalent 444 (calculated 445). The melting point of the methanol / ether recrystallized product is 197-198 ° ^ ·

The dicyclohexylamine salt from step b) (3 g, 6.72 mmol) is treated in a separatory funnel with 67.2 mL of 0.2 N aqueous sulfuric acid and 67 mL of ethyl acetate. The aqueous phase is further extracted with 60 ml and 30 ml portions of ethyl acetate, and the combined ethyl acetate solutions are washed with 15 ml portions of water and 15 ml of brine. The ethyl acetate solution is dried to Ν32δ0 ^: 1ΐ3, after which the solvent is evaporated in vacuo to give 1.9 g of crude product. The semi-solid product is dissolved in CH 2 Cl 2 and dispensed into ampoules, after which the solvent is removed as completely as possible by concentrating the product in a stream of nitrogen, followed by final drying under high vacuum. The product obtained is homogeneous by analytical thin layer chromatography: = 0.47 on silica gel, CHCl 3: MeOH: HOAc (12: 2: 1).

Example 12 DL-4- (3-mercapto-1-oxopropyl) -3-morpholinecarboxylic acid a) DL-4- (3-acetylthio-1-oxopropyl) -3-morpholinecarboxylic acid 0.0207 mol of 3-morpholinecarboxylic acid hydrochloride is reacted with 0.023 with 3 moles of 3-acetylthiopropionyl chloride in 50 ml of water containing 2.2 g of sodium carbonate at ice bath temperature. The pH of the reaction mixture is maintained at 8-8.2 by adding a total of 17 ml of 25% aqueous sodium carbonate solution during the reaction. When the pH ceases to change, the reaction is quenched with hydrochloric acid, and the reaction mixture is extracted with ethyl acetate. Extraction gives 5.0 g of crude syrupy product which is converted into the dicyclohexylamine salt in 50 ml of ethyl acetate, yield 7.1 g, m.p. 182-185 ° C (sintered at 170 ° C). Crystallization of the product from 300 ml of acetonitrile gives 5.7 g of a colorless salt, m.p. 187-189 ° C.

Analysis for C 18 H 18 N 2 O 5 S · C 2 H 2 N Calculated: C 59.70, H 8.65, N 6.33, S 7.25 Found: C 59.46, H 8.53, N 6.22, S 7.53.

14 67378

The dicyclohexylamine salt is converted to the free acid using 50 ml of 10% aqueous potassium bisulfate solution, m.p. 110-112 ° C.

Analysis for the compound

Calculated: C 45.96, H 5.79, N 5.36 Found: C 46.26, H 6.01, N 5.51.

b) DL-4- (3-mercapto-1-oxopropyl) -3-morpholinecarboxylic acid 0.013 mol of the product obtained from step a) are hydrolysed with 8 ml of concentrated ammonium hydroxide in 19 ml of water for 2 hours at room temperature with stirring. The colorless product obtained weighed 2.75 g.

Analysis for C 9 H 18 NO 2 S.1 / 4 t 2 O.

Calculated: C 42.94, H 6.08, N 6.26, S 14.33 Found: C 42.84, H 6.22, N 6.30, S 14.12.

Example 13 3- (3-Mercapto-2D-methyl-1-oxopropyl) -L-4-thiazolidinecarboxylic acid 1,1-dioxide 3- (3-acetylthio-2D-methyl-1-oxopropyl) -L-4 -thiazolidinecarboxylic acid 1,1-dioxide (2.9 g) is added with vigorous stirring to a mixture of 100 ml of water, 30 ml of methanol and 100 ml of concentrated ammonium hydroxide cooled in an ice bath. After 5 minutes, the reaction mixture is washed vigorously with 150 ml of ethyl ether and 50 ml of ethyl acetate and then acidified to pH 1 with concentrated hydrochloric acid. The mixture is extracted with ethyl acetate and the oily product from ethyl acetate is dissolved in 50 ml of ethyl acetate and treated with 1.3 ml of dicyclohexylamine and diluted with ether to give 2.1 g of product as the dicyclohexylamine salt, m.p. 221-222 ° C, LoC 1D = -71.4 ° (c = 1, in methanol).

On neutralization, 1.11 g of the free acid are obtained from the dicyclohexylamine salt as a hygroscopic, semi-solid.

Analysis for CQH2NO2S.1 / 2 Ho0 o -L J j 2 2

Calculated: C 34.77, H 5.10, N 5.07, S 23.21 Found: C 34.92, H 5.16, N 9.85, S 23.50

Neutralization equivalent, calculated 276.3, found 279.

15 67378

Example 14 DL-3- (3-Mercapto-1-oxopropyl) -2-thiazolidinecarboxylic acid and dicyclohexylamine salt a) DL-3- (3-Mercapto-1-oxopropyl) -2-thiazolidinecarboxylic acid dicyclohexylamine salt

Argon is introduced into 40 ml of water for 5 minutes. To this is added 35 ml of a 58% aqueous solution of NH 4 OH, and the mixture is stirred vigorously under argon while adding 7.00 g (15.7 mmol) of DL-3- (3-acetylthio-1-oxopropyl) -2-thiazolidinecarboxylate. the silicic acid-dicyclohexylamine salt in portions in solid form. The reaction mixture is stirred at room temperature for 20 minutes, then cooled to ice bath temperature and extracted with 40 ml and 30 ml portions of Cl 2 Cl 2. The aqueous phase is then rapidly recooled under argon and acidified to pH 1 with concentrated hydrochloric acid and then extracted with 100 ml and 3 x 50 ml of ethyl acetate. The organic solutions are combined and washed with 25 mL of water, 25 mL of brine, dried over Na 2 SO 4 and concentrated in vacuo to give a crude oily product, the oil is dissolved in 30 mL of methanol and treated with 3.13 mL (15, 7 mmol) dicyclohexylamine. To the mixture is added 250 ml of ether and allowed to stand at room temperature for 3 hours to give 5.71 g (90% yield) of the dicyclohexylamine salt, m.p. 206-210 ° C (dec.), Sintered at 200 ° C.

Analysis for .DCHA salt

Calculated: C 56.68, H 8.51, N 6.96, S 15.93 Found: C 56.41, H 8.37, N 6.80, S 16.10.

SH titration: 99.3%.

b) DL-3- (3-mercapto-1-oxopropyl) -2-thiazolidinecarboxylic acid

The dicyclohexylamine salt (5.60 g, 13.9 mmol) is added to a separatory funnel containing 100 mL of ethyl acetate and 140 mL of ice-cold, 0.3 N N 2 SO 4 solution flushed with argon. The layers are separated and the aqueous layer is further extracted with three 50 mL portions of ethyl acetate. The combined ethyl acetate extracts are washed with 25 mL of water and the brine is dried (Na 2 SO 4, under argon) and then concentrated in vacuo. Residual ethyl acetate is removed by dissolving the oily concentrate three times in about 150 67378 in 16 ml of CH10 and evaporating the solutions to dryness.

2

Analytical thin layer chromatography: Rf = 0.37, silica gel, EtOAc: pyridine: MeOH: H 2 O (60: 20: 6: 11).

Claims (1)

A process for the preparation of novel therapeutically active thiazolidine and thiazanecarboxylic acids of the formula I and their basic salts, R (R-CH) (CH-R 1) I | -5 2 | m | 1 R 4 - s - (CH 2) p --CH - CO - N-CH - CO - OH wherein R 1 and R 2 represent hydrogen or lower alkyl; R 1 is hydrogen or lower alkyl; R 1 is hydrogen, lower alkanoyl or benzoyl; X is O, S, SO or SO 2, wherein m is 2 and n is 1 when X is 0; m is 1 or 2, n is 0, 1 or 2 and m + n is 2 or 3; p is 0 or 1; characterized in that a) a compound of formula II / x \ (R_-CH) (CH-R1) 2. m | 1 n HN-CH - COOH 11 is acylated in one step by reacting it with a compound of formula III or a reactive equivalent thereof, ΐ 3 R 4 - S - (CH-,) - CH - COOH IIT to give a compound of formula I, or b) The compound of formula II is gradually acylated by reacting it with a haloacyl halide of formula IIIa f3 shark- (CHO) -CH-CO-hal (Ula) '* Γ 18 67378 wherein sha represents halogen to give the product of formula IIIb, R CH) (CH-R1) (IIIb) | hal- (CH2) p-CH-CO - N - CH-CO-OH which is then reacted with thiol-R4-SH to give the product of formula I, followed by possible hydrolysis the resulting compound of formula I wherein R 1 is other than hydrogen to a compound of formula I wherein R 1 is hydrogen and / or optionally oxidizing a compound of formula I wherein X is S to a compound of formula I wherein X is SO or SO 2, or a compound of formula I wherein X is SO, is oxidized to a compound of formula I wherein X is SO 2, and the compound of formula I thus obtained is optionally converted into a basic salt. 19 67378 For the preparation of a therapeutically active thiazide-lid- and thiazanecarboxylic acid with the formula X and a basic salt,? 3> I R. - S - (CH,) - CH - CO - N -CH - CO - OH 4. pi vilken formel R. | and R2 is an alkyl or alkyl alkyl; R 1 is a hydrogen or alkyl; do not claim alkanoyl or benzoyl; X is 0, S, SO or SO 2, color m 2 is 2 and n is 1, then X is 0; m is 1 or 2, then 0, 1 or 2 and m + n is 2 or 3; p är 0 or 1; In the case of HN-CH - COOH II and that the genomic form of a compound of formula ITI or and the reactive equivalents of R3 I3 R4 - S - (CH2) - CH - COOH III for use in the formulation of Formula I, or b) acylation of the formulation of Formula II with the addition of halogenacyl halides of the formulation purple