DD290331A7 - CYTIDINE-5'-DIPHOSPHATE-1-0-HEXADECYL-1-2-0- (2,2,2-TRIFLUORETHYL) GLYCEROL AND METHOD FOR ITS PREPARATION - Google Patents

Abstract

The invention relates to a process for the preparation of * and their salts. The goal is to develop a synthesis for the preparation of these compounds. According to the invention, the compound is synthesized by reacting cytidine-5-phosphate or a diphosphate-forming derivative of this compound with *.

Description

Field of application of the invention

The invention relates to a process for the preparation of cytidine-5'-diphosphate-1-0-hoxadecyl-2-0- (2,2,2-trifluoroethyl) glycerol and its salts. The connection is new. It is an analog of cytidine diphosphate diglyceride, which is important as a metabolite of the cellular biosynthesis of phospholipids. Field of application of the invention is the chemical and the pharmaceutical industry.

Characteristic of the known state of the art

It is known that nucleoside diphosphate diacylglycerol adducts can be prepared by reacting phosphatidic acids with nuklooside monophosphates (Paulus et al., J. Biol. Chom., 235 (1960) 1307; Chung Il Hong et al., J. Mod. Chem., 31 (1980) 1793). Methods for the preparation of nouen cytidine-5'-diphosphaM-0-alkyl-2-0- (2,2,2-trifluoroethyl) glycerol of the formula I are not known.

Object of the invention

It is the object of the invention to develop a process for the preparation of cytidine-5'-diphosphate-1-O-hexadecyl-2-0- (2,2,2-trifluoroethylene glycerol.

Explanation of the nature of the invention

The object of the invention is to develop a process for the preparation of novel, physiologically active compounds. According to the invention, the cytidine 5'-diphosphate-1-O-alkyl-2-0- (2,2,2-trifluoro-ethyl) glycerol is prepared by reacting 1-0-hexydecyl-2-0- (2,2,2 trifluoroethyl) glycero-3-phosphoric acid or the salts of the compound with cytidine-5'-phosphate or an activated derivative of the compound in the presence of a condensing agent such as NN-dicyclohexylcarbodiimide at temperatures between 15 ° C and 60 ° C, preferably at 35 ⁰ C. until 4O ⁰ C react. A preferred variant of the reaction consists in using cytidine B'-morpholidopnosphate as the reactive component and reacting it with the glycerophosphoric acid derivative. The reaction is generally carried out in the presence of an organic base, for example 4-dimethylaminopyridine or pyridine, it also being possible to work with the latter without further addition of a solvent. It is expedient to use an excess of the reaction component, wherein the molar ratio between cytidine 5'-morpholidophosphate and the 1-0-hexadecyl-2-0- (2,2,2-trifluoroethyl) glycero-3-phosphoric acid as 1: 0.5 to 2. The optimum reaction temperature is 35 ⁰ C to 4O ⁰ C. The reaction is carried out in the absence of water. After completion of the reaction, the reaction mixture is evaporated in vacuo to dryness. The remaining residue is dissolved in chloroform-methanol-verd. HCI recorded and the aqueous phase adjusted to a pH of 2 to 3. The chloroform-methanol phase obtained after phase separation is washed neutral with water, filtered and concentrated in vacuo. The residue obtained, after being taken up in chloroform-methanol, is treated with methanolic ammonia. The resulting diammonium salt is extracted with water from the chloroform-methanol solution. By concentrating the aqueous-alcoholic solution, taking up the residue obtained in a little methanol and adding dried acetone, the diammonium salt is obtained as a chromatographically uniform compound. The new compound is characterized by its cytostatic activity. Thus, it inhibits concentration-dependent cell proliferation of Ehrlich ascites tumor cells (Table 1).

Table 1

Inhibition of cell proliferation of Ehrlich ascites tumor cells by cytidine 5'-diphosphate-1-O-hexadecyl-2-0- (2,2, -trifluoroethylene glycerol

Concentration (pMolar) 3 10 30 100

Inhibition (%) 9.5 28 81 100

The invention will be illustrated by the following example.

Ausführungsbeljplel Cytldin-S 'dlphosphat-i-O-hexadecyl ^ -O-tf ^^ - trlfluorethyDglyceroll

A mixture of iO-hexadecyl-O-tf ^^ -trlfluoroethylglycorophoephoraauro (0.12 g, 0.25 mmol) and cytidyl-5'-morpholactophosphate (4-morpholene-N, N'-dlcyclohexylcarboxamldin salt) (0.30 g, 0.44 mmol) is rotary evaporated four times with in each case 5 ml of pyridine (with CaH!) Under high vacuum to dryness. The residue is then taken up again in 6 ml of pyridine and the solution with exclusion of moisture for 9 days at 37 ⁰ C stirred. Thereafter, the reaction mixture is concentrated to dryness in vacuo, the residue is pre-mixed several times with about 4 ml Toiuen and brought to Trockno ornout, the substance obtained in jeinom mixture of chloroform-methanol (1: 1, v / v, 30ml) and 0 , 01 N HCI (15ml) added. By dropwise addition of 1N HCl, the mixture is adjusted to a pH of 2 to 3. The chloroform-methanol phase is separated off, washed neutral with water and concentrated by filtration. The residue is dissolved in a mixture of chloroform (13 ml), methanol-water (2: 1, v / v, 10 ml) 1N methanol. NH ₃ (0.2ml) added. The organic phase is separated and washed twice with the mentioned Mothanol-Wassor-methanol. NH ₃ mixture (about 8 ml each) extracted. The combined aqueous phases were concentrated. The residue is crystallized with a little methanol and then washed several times with dry acetone. After drying, 30 mg of the dlammonium salt of I were obtained as a chromatographically uniform substance.

Thin-layer chromatography (finished plate Kieselgel 60, Merck):

Ri 0.39 (n-propanol / 25% w / w NH ₃ / water, 8: 2: 1, v / v / v; Ri 0.30 (chloroform / methanol / glacial / water, 25: 15: 2) : 4, v / v / v / v) UV absorption: X _m " 275.5 nm (water); X _m ," 282 nm (0.01 N HCl).

Fast atom bombardment mass spectrometry: m / z 783 (M)

m / z 558 ICeH ₃ JOCHI-CH (OCHJrF ₃ ) CHr-OP (O) -OP (O) -OH],

OH OH

m / z 540 ICUH ₃₃ OCHJ-CH (OCHjCF ₃ ) CHj-OP (O) -OP (OH,

OH O-

m / z 477 (CeH ₃₃ OCHj-CH (OCHjCF ₃₎ CH ₁ -OP (O) -O-),

OH

m'z CHD (M ^ H-ICH ₃₃ OCHJ-CH (OCHjCF ₃ ) CHj-O-]).

Claims (4)

1. A process for the preparation of Cytidinyl 5'-diphosphate-1-O-hexadecyl-2-0- (2,2,2-trifluoroethyl) glycerol and its salts, characterized in that 1-0-hexadecyl-2 -0- (2,2,2-trifluoroethyl) glycero-3-pho8phorsäure or its salts, with cytidine G'-phosphate or an activated derivative of this compound, in the presence of a condensing agent or an organic base at temperatures between 15 ⁰ C and , 6O ⁰ C, condensed and the resulting condensation product is isolated and optionally converted to a salt. 2. The method according to claim 1, characterized in that reacting cytidine-5'-morpholidophosphat in the presence of an organic base at temperatures of 35 ° C-40 ° C, wherein one of the components used in excess. 3. The method according to claim 1 and 2, characterized in that reacting the cytidine 5'-phosphate in the ratio 1: 0.5 to 2. 4. Cytidine 5'-diphosphate-1-O-hexadecyl-2-0- (2,2,2-trifluoroethyl) glycerol. "