DD242404A1 - PROCESS FOR PREPARING ALPHA-SULFONYLAMINOACYLATED 3-AMIDINO-PHENYLALANINESTER - Google Patents

Abstract

These compounds can be used as factor Xa inhibitors of high specificity in the laboratory diagnostic determination of the activity of prothrombin, via a reaction of the thrombin formed with chromogenic substrates, which they do not appreciably affect, or as anticoagulants. It is to be achieved to produce Na-sulfonylaminoacylated 3-Amidinophenylalaninester in the required purity and amount advantageous. This is done by reacting 3-cyanobenzylacylaminomalonate diester to give 3-cyanophenylalanine as the major product. After sulfonylaminoacylation, the thioimidic acid ester salts are obtained via the thioamides, which give the betaines of the Na-sulfonylaminoacylated 3-amidinophenylalanines with ammonium acetate, from which the corresponding esters are obtained with aliphatic or araliphatic alcohols in the presence of p-toluenesulfonic acid.

Description

-2- Explanation of the nature of the invention

The invention has for its object to provide a synthesis route for the preparation of Nasulfonylaminoacylierten 3-Amidinophenylalaninestern the structure I, which is technically easy to implement and disadvantages of previously known synthesis method substantially reduced or eliminated.

-CH ₂ -CH-COOR '

NH - CO - (CH,) - NH - SCy - T * ^Λ

In formula I, η = 1 to 5, R ¹ = alkyl, aryl or heteroaryl and R ² = alkyl or aralkyf.

The procedure of the synthesis is that a 3-Cyanbenzylhalogenid is reacted with a Acylaminomalonsäurediester, which is treated under reflux with glacial acetic acid and 3 N HCl in a mixing ratio 1: 2. 3-Cyanophenylalanine (structure II) is obtained as the main reaction product, in addition compounds of structure III and IV are obtained, which can be largely removed by titration with leaches to the isoelectric point of II, which thereby precipitates.

N C

HCCCv.

(V CH ^ - CH-CCCH

C HH -C _> ^

(V Ch ₂ - Ch- Ccoh

After sulfonylaminoacylation, compounds of the formula V in which η = 1 to 5 and R = alkyl, aryl or heteroaryl, are obtained which, after addition of hydrogen sulfide to the cyano group in pyridine / triethyiamine, give the thiocarbox-

/ V CH, -CH-COOH

amido compounds of the structure VI, where η and R have the abovementioned meaning.

Il

H ₂ N-Cn

/ "V-CH ₂ -Ch-COG h

These compounds are reacted with an alkyl halide to give trnoimidic acid esters of structure VII.

• HX'7A-Cn

VCH ₂ -CH-COOH

In this case, η = 1 to 5, R ¹ = alkyl, aryl or heteroaryl, R ² = alkyl and X is the meaning of the halogen atom of the eincjesetzten

From the thioimidic acid ester salts, the 3-amidine carboxylic acids of structure VIII are obtained by reaction with ammonium acetate, which crystallize out as betaines in pure form from the reaction mixtures.

NH - CO - (Ch ₂ ) -

η and R have the meaning analogous formula V.

According to the invention, the betaines obtained in this or another way are reacted with aliphatic or araliphatic alcohols in the presence of mineral acids or strong organic acids, in particular p-toluenesulfonic acid, to give the Na-sulfonylaminoacylated 3-amidinophenylalanine esters of structure I. The procedure according to the invention has the advantage that the crystalline compounds obtained as betaines of 3-amidinecarboxylic acids (structure VIII) can be reacted in one step with aliphatic or araliphatic alcohols in the presence of p-toluenesulphonic acid to give the 3-amidinecarboxylic acid esters of structure I which are described in US Pat pure form incurred.

Example 1:

10.0 g of 3-Cyanbenzylbromid, 11.0g Acetaminomalonsäurediethylester and 2.5g Kl are abs in 100ml. Dioxane at room temperature while stirring with a 1,15g sodium and 50 ml abs. Ethanol produced ethylate solution implemented. After five hours of stirring on the boiling water, the filtrate of the resulting suspension is mixed with plenty of water, the precipitate is filtered off with suction and recrystallized from methanol.

M.p .: 143-145 ⁰ C.

12.0 g of diethyl 3-cyanobenzylacetaminomalonate are dissolved in a mixture of 32 ml of glacial acetic acid and 64 ml of 3N HCl for 6 hours.

heated to reflux. I is distilled. Vak. most of the solvent. The residue is dried on the clay plate and reprecipitated from methanol / ether. For the substantial separation of the 3-carboxyphenylalanine from the hydrolysis product, 6 g of the hydrolysis product are dissolved in 20% methanol and titrated potentiometrically to pH 6.8-7.0. It crystallizes from 3-Cyanphenylalanin, which contains only a little of the 3-carboxy compound.

1.9 g of 3-cyanophenylalanine are dissolved in 21 ml of 1 N NaOH and shaken for 10 hours with a solution of 2.73 g of tosylglycyl chloride in 50 ml of ether. After separating the ethereal phase, the aqueous alkaline phase is extracted by shaking with ether. After acidification of the aqueous phase, Na (tosylglycyl) -3-cyanophenylalanine precipitates. It is filtered off with suction and crystallized from aqueous methanol. M.p .: 147-148 ⁰ C.

1.5 g of Na (tosylglycyl) -3-cyanophenylalanine are dissolved in 10 ml of pyridine and 2 ml of TEA. The batch is saturated with H ₂ S and stored for 36 hours at room temperature. The mixture is then stirred into hydrochloric acid ice water, wherein the thioamide is obtained in greasy form. It is taken up in ethyl acetate and the solution washed with dilute HCl and water. After drying the organic phase (Na ₂ SCt), the solvent is i. Vak. distilled off and further processed the amorphous residue in the resulting form.

1,1g thioamide are dissolved in 10 ml abs. Dissolved ethanol, the solution with 2.5 ml of methyl iodide and the mixture heated for 15 min. Reflux. After cooling, ether is added to give the thioimidic acid ester hydroiodide in a greasy form. The amorphous product obtained after precipitation from acetone / ether is further processed in this form. 0.9 g of thioimidic acid ester hydroiodide are dissolved in 15 ml of abs. Dissolved methanol, the solution with 0.2 g of ammonium acetate and the mixture heated for 2.5 hr. At 6O ⁰ C in a water bath. Upon cooling, Na (tosylglycyl) -3-amidinophenylalanine crystallizes out as betaine. M .: 267-269 ° C.

0.2 g of Na O -sylglycyD-3-amidinophenylalanine and 0.12 g of p-toluenesulfonic acid · H ₂ O are dissolved in 5 ml of abs. Dissolved methanol and the solution heated under reflux for 2.5 hours. After cooling NcHTosylglycyD-S-amidinophenylalanine methyl ester-tosylate is precipitated with ether and reprecipitated from acetone / ether. Mp .: from 100 ⁰ C.

Example 2:

1.9 g of 3-cyanophenylalanine (see Example 1) are dissolved in 21 ml of 1 N NaOH and shaken for 10 hours with a solution of 3.0 g of β-naphthylsulfonylglycyl chloride in 40 ml of ethyl acetate. After separating the organic phase, the aqueous phase is acidified with dilute HCl, whereby Na (2-Naphthylsulfonylglycyl) -3-cyanphenylalanine precipitates. After aspirating is recrystallized from aqueous methanol.

Mp .: 161 ° C

2.0 g of Na- (2-naphthylsulfonylglycyl) -3-cyanophenylalanine are dissolved in 20 ml of pyridine and 4 ml of TEA. The batch is saturated with H ₂ S and stored for 48 hours at room temperature. The mixture is then stirred into hydrochloric acid ice water, the thioamide extracted with ethyl acetate and the solvent after drying the organic phase (Na ₂ SO ₃ ) i. Vak. distilled off. The amorphous residue is further processed in the resulting form.

2.8 g of thioamide are dissolved in 20 ml of acetone, the solution is mixed with 5 g of methyl iodide and the mixture is heated under reflux for 15 min. After cooling, ether is added to give the thioimidic acid ester hydroiodide in a greasy form. The amorphous product obtained after precipitation from acetone / ether is further processed in this form.

3.1 g of thioimidic acid ester hydroiodide are dissolved in 40 ml of methanol, the solution is admixed with 1.0 g of ammonium acetate and the mixture is stirred for 2.5 h. warmed at 6O ⁰ C in a water bath, after about 30 min., The betaine of Na (2-naphthylsulfonylglycyl) -3-amidinophenylalanine begins to crystallize. After cooling, the mixture is filtered off with suction and recrystallized from water / ethanol 9: 1. M.p .: 264-266 ° C.

0.2 g of Na (2-naphthylsulfonylglycyl) -3-amidinophenylalanine and 0.11 g of p-toluenesulfonic acid · H ₂ O are in 5ml abs. methanol

From Na (tosylglycyl) -3-amidinophenylalanine and Na (2-naphthylsulfonylglycyl) -3-amidinophenylalanine, the following compounds were obtained according to Examples 1 and 2:

connection

mp .:

reaction conditions

reaction time

Na (Thosylglycyl) -3-amidinophenyl from 90 ⁰ C Heating under reflux 2.5 hours alanine tosylate Na (tosylglycyl) -3-amidinophenyl from 80 ⁰ C Heating under reflux 20 min. alanine n-butyl ester tosylate Na (tosylglycyl) -3-amidinophenyl from 95 ⁰ C Heating under reflux 15 hours alaninisopropyl ester tosylate Na (tosylglycyl) -3-amidinophenyl from 90 ⁰ C Heating on the boiling 2 hours. alanine benzyl ester tosylate water bath Na (tosylglycyl) -3-amidinophenyl from 85 ⁰ C Heating on the boiling 2 hours. alanine (2-phenyl) ethyl tosylate water bath Na (2-naphthylsulfonyl-glycyl) - from 100 ⁰ C Heating under reflux 2.5 hours 3-amidinophenyl-alaninethylester- tosylate Na (2-naphthylsulfonyl-glycyl) - from 80 ⁰ C Heating in a water bath 5 hours 3-amidinophenyl-alanine n-butyl ester 60 ° C tosylate Na (2-naphthylsulfonyl-glycyl) - AB100 ° C Heating under reflux 15 hours 3-amidinophenyl-alaninisopropylester- tosylate Na (2-naphthylsulfonyl-glycyl) - ab95 ° C Heating in a water bath 5 hours 3-amidinophenyl-alaninbenzylester- 60 ° C Na (2-naphthylsulfonyl-glycyl) - from 90 ⁰ C Heating in a water bath 5 hours 3-amidinophenylalanin- (2-phenyl) ethyl 60 ° C ester tosylate

Claims (1)

Invention claim: A process for the preparation of Na-sulfonylaminoacylated 3-amidinophenylalanine esters of general formula I by reacting a 3-cyanobenzyl halide to give a 3-cyanobenzyiacylaminomalonic acid diester reacting in a mixture of glacial acetic acid and HCl in the presence of S-cyanophenylalanine hydrochloride, 3-carboxamidophenylalanine hydrochloride and Carboxyphenylalanine hydrochloride results, which are largely separated by isoelectric precipitation, the further reaction of the thus purified 3-Cyanphenylalanins with a sulfonylated amino acid halide to a Nasulfonylaminoacylierten 3-cyanophenylalanine, which is reacted with H ₂ S in pyridine / triethylamine to Thiocarboxamidoverbindung after reaction with a Alkyl halide provides a Thioimidsäureesterhalogenid, characterized in that is obtained by heating with ammonium acetate in alcoholic solution, the betaine of a Nasulfonylaminoacylierten 3-Amidinophenylalanins, from which with aliphatic or aralipha Tischen alcohols in the presence of p-toluenesulfonic acid, the tosylate of a Na-sulfonylaminoacylated 3-Amidinophenylalaninesters arises, the general formula -CH-CQOR ² - GO- (CH,) - NH-SO, - "^ has, in which η = 1 to 5, R ¹ = alkyl, aryl or heteroaryl and R ² = alkyl or aralkyl. Field of application of the invention The invention relates to a process for the preparation of Na-alkyl, Να-aryl or Na-heteroarylsulfonylaminoacylated 3-Amidinophenylaianinestern. These compounds are factor Xa inhibitors of high specificity and can be advantageously used to eliminate the activity of factor Xa in the determination of prothrombin mitchromogenem substrate in biological materials or as direct anticoagulants. Characteristic of the known technical solution Inhibitors of coagulation factor Xa are potential anticoagulants (J.Hauptmann, F.Markwardt and P. Walsmann, Thromb. Res., 12,735, 1978, R. R. Tidwell, W. P. Webster, F. R. Shaver and J. D. Geratz, Thromb. Res., 19, 399, 1980). Relatively specific inhibitors of Factor Xa have been found among 3-amidinophenyl derivatives (J.Stürzebecher, F. Markwardt and P. Walsmann, Thromb. Res. 9,637,1976, Thromb. Res. 17,545,1980). However, for inhibitors that are to be used as anticoagulants, a differentiation between inhibition of factor Xa and other serine proteinases of the blood, which are involved in the clotting ah such. As thrombin, not be sought. Interest in specific inhibitors of factor Xa, which in particular does not affect the activity of thrombin, exists for laboratory diagnostic methods for the determination of clotting factors with the aid of chromogenic substrates. Since the chromogenic peptide substrates are not absolutely specific, they are also cleaved by enzymes that are active in the blood plasma in addition to the enzyme to be determined or are activated during the determination. The activity of factor Xa is particularly troublesome in the determination of prothrombin mitchromogenem substrate (BRJKirchhof, C. Vermeer and HCHemker, Thromb. Res. 13,219.1978, LiII, prostaglandins and platelet function, ed. Breddin, Schattauer-Verlag, Stuttgart 1978, 279). In this method of determination, the prothrombin is activated in the test plasma by added factor Xa and the thrombin formed by its activity against chromogenic substrate detected. Since the added factor Xa also causes cleavage of the substrate, its elimination by a specific inhibitor is necessary for the precision of the method. The previously known as relatively specific inhibitors of factor Xa derivatives of 3-Amidinobenzamidins do not have the high specificity for said diagnostic method. The developed derivatives of Naarylsulfonylaminoacylierten 3-Amidinophenylalanins described have the specificity of the inhibitory effect required for the described use. Method for Preparation Na-arylsulfonylaminoacylated amidinophenylalanine ester (G.Wagner, B. Voigt and H.Vieweg, Pharmaceutics 39,226 [1984], G.Wagner, B. Voigt, H.Vieweg, F.Markwardt and J. Falling Becher, DD-PS 155954 ), while in principle allow to produce substances of the desired type, but these methods are complicated or less suitable because run under the necessary conditions side reactions. The resulting in moderate yields end products are difficult to clean. Object of the invention The aim of the invention is to produce inhibitors of factor Xa from the class of nasulfonylaminoacylated 3-amidinophenylalanine esters in an economically advantageous and technically simple manner. - -