DD277462A1 - PROCESS FOR THE PREPARATION OF CYTOSINARABINOSIDE 5'-DIAMIDOPHOSPHATES - Google Patents

Abstract

The invention relates to a process for the preparation of cytosine arabinoside-5-diamidophosphates of the general formula I. Fields of application of the invention are the chemical and pharmaceutical industry. The compounds are obtained by phosphorylation of Cytosinarabinosid and subsequent reaction of the resulting Cytosinarabinosid-5-dichlorophosphates with ammonia or amines. They have cytostatic activity. Formula I

Description

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reacts, this reacts with ammonia or an amine, such as methylamine, dimethylamine or morpholine and the compound I in a known manner: isolated and purified.

2. The method according to claim 1, characterized in that phosphorylated with phosphorus oxychloride in a molar ratio of 1: 1.5 to 2.2 in the presence of trimethyl or triethyl phosphate anhydrous and under ice cooling.

3. The method according to claim 1, characterized in that reacting the compound II with a highly concentrated aqueous ammonia or amine solution in excess.

Field of application of the invention

The invention is directed to a process for the preparation of novel cytosine arabinoside B'-diamidophosphates. Areas of application of the invention are the chemical and pharmaceutical industry.

Characteristic of the known state of Tekhpik

It is known that nucleoside-5'-diamido can be synthesized by the reaction of nucleoside S'-dichlorophosphates with amines (S. Bottka and J. Tomasz, Tetrahedron 35 [197912909-2912, J. Tomasz, J. Carbohydrates, nucleosides, nucleotides 8 [1981] 557-572). Cytosine arabinoside S'-diamidophosphates of general formula I are hitherto unknown.

Object of the invention

It is the object of the invention to develop a process for the preparation of the cytosine arabinoside 5'-diamidophosphates of general formula I.

Explanation of the essence of the invention

According to the invention, the cytosine arabinoside-α'-diamidophosphates of the general formula I,

in which R has the meanings already mentioned, prepared by reacting cytosine arabinoside with a suitable phosphorylating agent, e.g. Phosphoroxychlorid, preferably in the molar ratio 1: 1.5 to 2.2, converted into the Cytosinarabinosid 5'-dichlorophosphate of the general formula II and this with ammonia,

Methylamine, dimethyiamine or morpholine brings to implementation. The conversion according to the invention of the cytosine arabinoside into the cytosine arabinoside S'-dichlorophosphate is generally carried out in such a way that to a mixture of the nucleoside with a trialkyl, preferably trimethyl or triethyl (about 1-3ml / mmol nucleoside) under anhydrous conditions phosphorus oxychloride Ice cooling and stirring is dropped. The reaction, which is conveniently followed by thin-layer chromatography, is complete after about 1 to 4 hours of stirring at 0-5 ° C. The reaction taking place under these reaction conditions essentially results in a substitution in the 5'-position of the nucleoside A clear concentrated solution of the reaction mixture is then added in concentrated form to a highly concentrated aqueous ammonia or amine solution with continuous stirring, and the mixture is then concentrated to dryness in vacuo.The crystalline residue is then taken up in water and brought to the pH with the appropriate ammonia or amine solution The aqueous solution of the crude cytosine arabinoside 5'-diamidophosphate obtained in this way is purified (separation of anionic impurities, NH ₄ HCO ₃ , unreacted starting material, PO (NH ₂ I _3, etc.) onto an anion exchange column, eg Dowex 1x8 (HCO ₃ "form), as eluent tel water is used. The fractions containing the final product are concentrated to dryness in vacuo and the resulting substance in case of need by recrystallization, z. B. from water, cleaned. The compounds of general formula I are novel and cytostatically active. They also represent potentially new transport and active forms of the era C, which has therapeutic significance as cytostatic and antiviral and are valuable intermediates in the preparation of nucleotide derivatives of the Ara-C type.

Table 1 lists the antiproliferative activity of the cytosine arabinoside 5'-diamidophosphate I (R = NH ₂ ) synthesized according to the invention in comparison with Ehrlich ascitic tumor cells as a function of the concentration of the active ingredient.

Table 1: Growth inhibition of Ehrlich ascites tumor cells in viiro under the influence of different concentrations of era C-5'-diamidophosphate I (R = NH ₂ ) after 48 hours

Concentration (μΜ) 100 30 10 3

Inhibition (%) *> 78.3 68.8 58.7 43

a) Average of 5 trials

The invention will be explained by the following example.

embodiment

Cytosine arabinoside 5'-diamidophosphate I (R = NH ₂ )

To a mixture of 468 mg (2 mmol) of cytosine arabinoside and 5 ml of freshly distilled trimethyl phosphate are added dropwise with stirring and ice cooling 240μΙ (2.64 mmol) freshly distilled phosphorus oxychloride. The mixture is stirred for 1 hour under ice-cooling. After this time, the resulting clear solution is mixed with 25 ml of a 7 M aqueous ammonia solution and then concentrated to dryness in vacuo. The residue is dissolved in 15 ml of water and treated with conc. aqueous ammonia solution adjusted to pH 8.5. This solution is placed on a Dowex 1 χ 8 column (HCO ₃ form, 3x 80 cm) and eluted with water. From fractions 19-26 (20 ml each), 560 mg (87.2%) of nucleoside diamidophosphate I are obtained, which is recrystallized from water for further purification.

TLC (Merck precast plates, diatomaceous earth 60 F-254, n-propanol / 25% aqueous ammonia / water, 8: 3: 2, v / v / v) R ₁ 0.45.

Elemental analysis calcd. WrC ₀ H ₁₆ N ₅ PO ₆ χ 1.5H ₂ O (348.26) C, 31.04, H, 5.49, N, 20.11; Found C 31.04, H 5.53, N 20.38 (%).

UV spectroscopy: A _mK 280 (0.01 N HCl). ³¹ P NMR (DMSO, F-) 18.92 ppm.

Claims (1)

1. A process for the preparation of cytosine arabinoside B'-diamidophosphates of the general formula I, R ° / - ^v
in which R = NH ₂ , NHCH ₃ , N (CH ₃ ) ₂ , 0 N, characterized in that cytosine arabinoside (Era C) with a Phosphoryiierungsmittel in the presence of a Trialkyiphosphates to Cytosinarabinosid 5'-dichlorophosphst of the general formula II