DD277463A1 - PROCESS FOR PREPARING CYTOSINARABINOSIDE 5'-MONOAMIDOPHOSPHATE - Google Patents

Abstract

The invention relates to a process for the preparation of cytosine arabinoside-5-monoamidophosphate of the formula I or its salts. Areas of application of the invention are the chemical and pharmaceutical industry. Compound I is obtained by partial hydrolysis under acidic reaction conditions and subsequent chromatographic purification from cytosine arabinoside 5-diamidophosphate. It has cytostatic activity. Formula I

Description

Explanation of the essence of the invention

According erfindungsgeqnäß the Cytosinarabinosid B'-monoamidophosphate of the formula I.

synthesized by partially hydrolyzing Cytosinarabinosid ö'-diamidophosphate of formula II by the action of aqueous acids at a constant pH.

The preparation according to the invention is generally carried out by adding an aqueous solution of sulfuric acid (diluted 1: 5) into a solution of diamidophosphate II in water at room temperature with stirring. It is essential that the sulfuric acid is added dropwise in such a way that the pH is kept constant at 1.9. Dio total reaction time is about 30 minutes. Subsequently, the reaction solution is adjusted to a pH of 7 to 8.5 with concentrated aqueous ammonia and placed on a DEAE-cellulose column. The elution is carried out with a linear gradient of an aqueous solution of triethylammonium bicarbonate (pH 7.5), the cytosine arabinoside 5'-monoamidophosphate I being obtained as a thin-layer chromatographically pure triethylammonium salt. Compound I is new and represents a potential transport and active form of the C-era used as cytostatic

 It has no significant cytostatic activity against tumor cells in vitro. Table I shows the inhibition of cell growth of Ehrlich ascites tumor cells as a function of the concentration of active ingredient I.

Table I: Inhibition of cell growth of Ehrlich ascites tumor cells in vitro upon exposure to varying concentrations of the ara-C-5'-monoamidophosphate I after 48 hours

Concentration (μΜ) 100 30 10 3

Inhibition (%) * 86.1 66.2 57.4 36.1

* Average values of 4 attempts

In addition, it is useful as a synthetic intermediate for the preparation of nucleotide derivatives

 The invention will be illustrated by the following example.

embodiment

Cytosine arabinoside 5'-monoamidophosphate (I)

560 mg of cytosine arabinoside-α'-diamidophosphate (II) (1.75 mmol) are dissolved in 30 ml of water. Dilute sulfuric acid (prepared from 1 volume of conc. H ₂ SO ₄ + 4 volumes of H ₂ O) is added dropwise to the solution over a period of 30 minutes to maintain the pH at the constant value of 1.9. Subsequently, the reaction solution with conc. adjusted to a pH of 7-8.5 aqueous NH ₃ , diluted with water to a volume of 200 ml and applied to a DEAE-cellulose (HCO ₃ ~ -form, column 3 x 80cm). The elution is carried out with a linear gradient of 0-0.2M triethylammonium bicarbonate in water (2 liters each).

The fractions 78-89 (fraction size 20.7 ml) are concentrated in vacuo, the residue is re-distilled 3 times with 20 ml of methanol and dried over P ₂ O ₅ . Yield of I (triethylammonium salt): 465.1 mg (63%). TLC (Merck precast plates, silica gel 60 F-254, n-propanol / 25% aq. NH ₃ / H ₂ 0.8: 3: 2, v / v / v) R, 0.52. Elemental analysis calculated PO x ₇ H ₂ O (441.43) C40.81, H7.31, N 15.86 for C ₁₅ H ₃₀ N _5; found C40.83, H7.01, N 16.08 (%)

UV spectroscopy: X _msx 280 (0.01 NH ₄ ), ³¹ P NMR (DMSO, δ) 8.78ppn>.

Claims (1)

A process for the preparation of cytosine arabinoside B'-monoamidophosphate of the formula I and its salts, characterized in that cytosine arabinoside 5'-diamidophosphate of the formula II under acidic conditions at a constant pH, subjected to partial hydrolysis and the
resulting compound I optionally converted by known methods into the salts.
2. The method according to claim 1, characterized in that an aqueous solution of sulfuric acid dropwise at a cytosine arabinoside 5'-diamidophosphatlösung
pH value of 1.9, then adjusts a pH of 7 to 8.5 and the compound of formula I isolated by methods known per se. Field of application of the invention The invention relates to a process for the preparation of a novel Cytosinarabinosid 5'-monoamidophosphates or salts thereof. Areas of application of the invention are the chemical and pharmaceutical industry. Characteristics of the known state. of the technique It is known that nucleoside 5'-monoamidophosphates can be synthesized by reacting nucleoside 5'-phosphoc with ammonia in a mixture of formamide / tert-butanol in the presence of dicyclohexylcarbodiimide (RW Chambers et al., J. Chem. Amer. Chem. Soc., 80 [1958] 3752). In the following years, a number of nucleoside 5'-monoamidophosphaten by
Hydrolysis of the corresponding dianidophosphates (J. Tomasz, Carbohydrates, Nucleosides, Nucleotides, 8 (1981) 557). The 5'-amidophosphate of cytosine arabinoside I is not yet known. Object of the invention It is the object of the invention to develop a process for the preparation of the cytidine-5'-monoamidophosphate.