DD289894A7 - METHOD FOR PRODUCING CYTIDINE 5'-DIPHOSPHATE-ALKYL-GLYCEROL ANALOGS AND SALTS THEREOF - Google Patents

Abstract

The invention relates to a process for the preparation of * and its salts. The compound is synthesized according to the invention by reacting cytidine-5-monophosphate as such or in the form of a reactive derivative of this compound with 1-0-hexadecyl-3-deoxy-3-chloroglycero-2-phosphoric acid.

Description

Field of application of the invention

The invention relates to a process for the preparation of cytidine B'-diphosphate-S-chloro-S-deoxy-i-O-hexadecylglycerol and its salts.

Field of application of the invention is the chemical or pharmaceutical industry.

Characteristic of the known state of the art

It is known that nucleoside phosphates can be condensed with diacyl glucophosphates in the presence of a condensing agent to give nucleoside diphosphate diacylglycerols (Paulus M. et al., J. Biol. Chem., 235 [1960] 1 307). Cytidine 5'-monomorpholidophosphates have also been used to prepare such adducts (B. Agranoff et al., Biochem., Prop., 10 (19631 47).) For cytidine S'-diphosphate-alkylglycerol type I analogs, no Bokannt manufacturing process.

Object of the invention

The object of the invention is to provide a method for producing the above-mentioned. Develop a connection.

Explanation of the essence of the invention

According to the invention, the cytidine-5'-diphosphate-3-chloro-3-deoxy-1-O-hexadecylglycerol is prepared by reacting cytidine-5'-monophosphate, preferably in the form of a reactive derivative, for. As the cytidine-o'-monomorpholidophosphates (as 4-morpholine-N, N'-dicyclohexylcarboxamidin-salt) with 1 -G-hexadecyl-S-deoxy-a-chloroglycero ^ -phosphof acid in Molverhältr.is of 1: 0 , 6 to 2 in the presence of 4-dimethylaminopyridine, pyridine or other inert organic base, optionally in the presence of a solvent for the reaction. The reaction is carried out between 20 and 6O ⁰ C, preferably at 35 and 4O "C, with stirring and exclusion of moisture. The reaction time is generally from 24 hours to 10 days. The hoaktion is aborted if the reactants used in deficiency chromatographically no longer The reaction mixture is worked up by taking up in chloroform-methanol-water, optionally after removal of solvent, pyridine or other volatile bases and separation of the aqueous phase after adjustment of the pH to 2 to 4. The obtainable by concentration of the organic phase free acid is purified as such or after conversion to the diammonium salt in a conventional manner, for example by column chromatography on silica gel The novel compound is an analogue of the cellular cytidine-α'-diphosphnt diglyceride and is physiologically active for example, to Ehrlich ascites tumor cells at one concentration of 3x 10 ~ ^s an inhibition of cell proliferation by an average of 40%. The invention will be illustrated by the following example.

embodiment

Cytidine-5'-diphosphate-3-chloro-3-deoxy-1-0-hexadecyglycerol

To a solution of 0,664g (1,6mmol) S-chloro-S-deoxy-iO-hexadecylglycero-i-phosphoric acid and 0,77g (6,3 r no!) 4-dimethylamino-anhydride in 10ml dried chloroform are added 0.882 (1, 3mmol) of cytidine 5'-monomorpholidophosphate (4Morpholine-N, N'-dicyclohexylcarboxamidine salt). The mixture is stirred in a sealed vessel at 35 ⁰ C with exclusion of moisture for 5 days. Thereafter, the reaction mixture is taken up in chloroform-methanol-water (pH 2), the organic phase is washed with water and evaporated in vacuo. The residue is taken up in chloroform-pyridine-formic acid (50: 30: 7, v / v / v) and separated by column chromatography on 30 g of silica gel 60 (Merck, 0.040-0.063 mm). The elution is initially carried out with 270 ml of the same solvent, then with chloroform-methanol (9: 1.340 ml, 8: 2.300 ml, each v / v).

The fraction of 31-55 (fraction size 20ml) was eingoongt in vacuum to Trockno. Dor obtained residue is taken up in oinom mixture of 28 ml of chloroform-methanol (1: 1, v / v) and 9ml 0.01 N HCl and the mixture is adjusted to pH 2 with 1N HCl. The phases are separated, the organic phase is again treated with 5 ml of chloroform-methanol (1: 1, v / v). The combined organic phases are dissolved with the addition of methanolic ammonia. The resulting diammonium salt of compound I is crystallized by treatment with methanol-dimethyl ether, washed with diethyl ether and dried. Yield: 0.340g. Thin Layer Chromatography (Merck Finished Plate Kieselgel 60 F 254): R, 0.42 (chloroform-methanol-glacial acetic acid-water 50: 30: 4: 8, v / v / v / v); R ₍ 0.42 n-propanol-25% aqueous ammonia-water 8: 2: 1, v / v / v)

US absorption: A _mlx 281.5nm (0.01 N HCI).

Claims (5)

1. A process for the preparation of cytidine-B'-diphosphate-S-chloro-S-deoxy-iO-hexadecylglycerol and salts thereof, characterized in that cytidine-5'-monophosphate or a roaklives derivative of this compound with "lO- Hexadecyl-S-deoxy-S-chloroglycero ^ phosphoric acid, preferably in the form of the salts of these compounds and in the presence of a condensing agent, condensed and the resulting condensation product isolated in a conventional manner and optionally converted into a salt. 2. The method according to claim 1, characterized in that the reaction is carried out in the presence of an inert organic base at temperatures of 20 to 60 ° C. 3. The method according to claim 2, characterized in that the reaction takes place at a temperature of 35 ⁰ C and 4O ⁰ C. 4. The method according to claim 1, characterized in that reacting cytidine-5'-monophosphate with the i-O-hexadecyl-S-deoxy-S-chloro-glycero ^ -phosphoric acid in a molar ratio 1: 0.6 to 2. 5. Cytidine-α'-diphosphate-S-chloro-3-deoxy-1-O-hexadecylglyceroi.