DD239208A1 - PROCESS FOR THE PREPARATION OF 1-O-ALKYL-2-O-TRIFLUORALKYLGLYCEROPHOSPHO-L-SERINES - Google Patents

Abstract

The aim of the invention is to develop a process for the preparation of 1-0-alkyl-2-0-trifluoroalkylglycero-3-phospho-L-serines. According to the invention, these compounds are prepared by reacting 1-0-alkyl-2-0-trifluoroalkylglycero-3-phosphoric acid esters in the presence of phospholipase D with L-serines.

Description

in the pure alkyl radical having 5 to 30 carbon atoms and η = 1, 2.3, and of their salts, characterized in that 1-0-alkyl-2-0-trifluoroalkyl-glycero-3-phosphoric acid ester of the general formula II, in the R and η the already mentioned

CH ₂ -OR

CH-O- (CH ₀₎ -CF., II

i. O

CH ₉ -OPOX

* · Ι

O"

And X is a CH _{3 group} or an alkyl radical having 2 to 6 carbon atoms, which may also be substituted by Cl, Br, OH, NH ₂ , NHCH ₃ .N (CH ₃ I ₂ or N (CH ₃ I ₃ , means reacting with L-serine under the catalytic influence of Phosphorlipase D and isolated the resulting glycerophosphoserin derivative according to methods known per se.

Field of application of the invention

The invention relates to a process for the preparation of i-O-alkyl-2-O-trifluoroalkylglycerophospho-L-serines of general formula 1, in which

CH ₂ -OR

CH-O- (CH ₀₎ -CF, I

, O ^{ά n ö}

CH ₂ -OPO-CH ₂ -CH-COOH

I (

OH NH ₂

R is an alkyl radical having 5 to 30 carbon atoms and η = 1, 2.3, and their salts.

The compounds represent analogues to the naturally occurring phosphatidylserines. They have an increased biostability compared to the latter, since they are not cleaved by the phospholipase A ₁ and A _2, for example. Field of application of the invention is the pharmaceutical or chemical industry.

Characteristic of the known technical solutions

Phosphatidylserines can be synthesized by condensation of phosphate acids, phosphatidic acid chlorides or phosphatidic acid ester chlorides with protected serine derivatives followed by cleavage of the serine protecting groups. Also, the reaction of diacylglycerol halohydrins with protected phosphoserines is known as a method of producing phosphatidylserines (AJ Slotboom et al., Chem. Phys. Lipids, 5 [1970] 310-397, H. Ebil, Chem. Phys. Lipids, 26 [1980] 405 -429; A. Hermetter et al., Chem. Phys. Lipids, 30 [1982] 35-45). Furthermore, they have also been obtained by enzymatically catalyzed reactions of other natural phospholipids, such as egg lecithin. Methods for the preparation of glycerophospho-L-serine type I are not known.

Object of the invention

It is therefore the object of the invention to develop a process which allows the preparation of type I glycerophospho-L-serines.

Explanation of the essence of the invention

According to the invention the iO-alkyl ^ -O-trifluoralkylglycero-S-phospho-L-serines of the general formula I are obtained by reacting 10-alkyl ^ -O-trifluoralkylglycero-S-phosphoric acid ester of the general formula II in which R and η the already

CH ₂ -OR

CH-O- (CrO-CF ,, II

¹ R

CH ₀ -OPOX

and X is a CH _{3 group} or an alkyl radical having 2 to 6 carbon atoms, which may also be substituted by Cl, Br, OH, NH ₂ , NHCH ₃ , N (CH ₃ I ₂ or N (CH ₃ J ₃₎ , means reacting with L-serine under the catalytic influence of phospholipase D. The enzyme can be obtained from readily available vegetable raw materials, for example from white cabbage, by methods known per se.

For the preparation of the compound of type I, a phospholipid type II with excess L-serine in an aqueous acetate or Tris buffer solution or suspension pH 4.8 to 8.5) containing calcium chloride (0.01 to 0.1 M) and the ether or a mixture of ether with further organic solvents, for example chloroform, and phosphoryl lipase D was added, at temperatures between 5 and 6O ⁰ C, preferably 40 ⁰ C, reacted with vigorous stirring. After a reaction time of 0.5 to 24 hours, a 0.1 M EDTA solution is added to deactivate the enzyme and the - resulting phosphatidylserine analogues isolated and purified by chromatographic methods. The compounds synthesized according to the invention are cancerostatic.

Thus, for example, shows that 1-0-hexadecyl-2-0- (2,2,2-trifluoro-ethylglycero-3-phospho-L-serine a 100% inhibition of growth of Ehrlich ascites tumor cells at concentrations <10 ~ ⁴ M ,

embodiment

1-0-hexadecyl-2-0- (2,2,2-trifluoroethyl) glycero-3-phospho-L-serine (Formula I: R = C ₁₆ H ₃₃ , η = 1)

a) To a solution of 1 g L-serine in 1.9 ml of 0.1 M acetate buffer (pH 5.6) with 0.1 M CaCl ₂ , 40 mg of 1-0-hexadecyl-2-0- (2,2, 2-trifluoroethyl) glycero-3-phosphocholine, (formula II: R = C ₆ H _33, n = 1, X = CH ₂ -CH ₂ - ^ (CH ₃ I _3), 2mitEther / chloroform (9: 1, v / v, ethanol free) and 100mg of a obtained from 500g cabbage phospholipase D preparation added. After 4 hours of stirring at 4O ⁰ C, the mixture is added at 20 C ⁰ with 4.35 ml of 0.1 M EDTA solution, the organic phase by evaporation of nitrogen and the aqueous phase is stirred with 4.3 times the volume of chloroform / methanol (5: 8, v / v) for 30 minutes, whereupon the organic phase is separated off and concentrated, and the residue obtained is added to 20 g of carboxymethylcellulose (Servacel CM52 After elution with 75 ml of chloroform (fraction 1), 500 ml each of chloroform / methanol (9: 1.8: 2.7: 3.1: 1 v / v, fractions 2 to 5), the glycerophosphpserine derivative is passed through Concentration of fraction 5 obtained in pure form (12 mg) TLC (Merck Kieselgel 60, finished plate): Rf 0.13 (CHClA / CHaOH / HjO, 50: 24: 4, v / v / v).

b) Reaction of 1 g L-serine and 40 mg of 1-0-hexadecyl-2-0- (2,2,2-trifluoroethyl) glycero-3-phosphonic acid 2-bromoethyl ester (formula II: R = C ₁₆ H ₃₃ , η = 1, X = CH ₂ -CH ₂ -Br) analogously to a). After working up and chromatographic separation of the reaction mixture analogously to a), 1.35 mg of the glycerophosphoserine derivative are obtained.

Claims (1)

Invention claim: 1. A process for the preparation of i-O-alkyl ^ -O-trifliioralkyl-glycero-S-phospho-L-serines of general formula I, CH ₉ -OR I ^ά CH-O- (CH ₀ ) -CFo I IO ^{ά n ö} " CH ₀ -OPO-CH ₉ -CH-COOH I OH NH ₀ C. ι