DE1922702A1 - Process for the preparation of 1-glycosyl-5-azacytosines - Google Patents

Description

24. a. 1969

Application files: 75/153

PATENT APPLICATION

Applicant; Ceskoslovenkso akademie ved _{# /} Praha 1

Title: Process for the preparation of l-glycosyl-5-azacytosines

The invention relates to a process for the preparation of l-glycosyl-5-azacytosines of the general formula I.

in which R is a glycoeyl, W is hydrogen or an alkyl

3 4

1-4 carbon atoms and R and R, identical or different, Hydrogen atoms, Al Icy le with 1-4 carbon atoms or azalkyls with 7-10 carbon atoms mean.

9008 * 7/1214

Two substances of the above type have significant biological effects, namely 1- | M) -ribofuranosyl-4-amino'- ⁷ 1 _/ 2-dihydro-1 _/ 3 _/ 5-triazin-2-one (5-azacytidine ) and the 1- (2-deoxy - ^ - D-ribofuranosyl) -4-amino-1 _/ 2-dihydro-1 _/ 3 _/ 5-tfiazin-2-one (the 5-aza-2 '-deoxycitidine).

The 5-azacytidine, a pyrimidine antimetabollite, retards the growth process of bacteria even in low concentrations and has a high antileukamic effect in mice. In the meristem of the V. faba it causes inhibition of mitoses and aberrations. In bacteria, 5-azacytidine has chromosomal mutagenic effects * Furthermore, it delays the formation of inductive enzymes in cells from mammals and the regeneration of rat liver after a repatectomy; on the other hand, it protects mice from the effects of X-rays.

The 5-aza-2-deoxysitin _/ similar to the 5-azacytidine, significantly dampens the induction of experimental leukemia and has marked bacteriostatic properties even in low concentrations.

It can therefore be assumed that some other 1-glycotyl-5-azacytosines will also have biological activity "

The preparation of l-glycosyl-5-azacytosines is the subject of DBR patents No. 1 244 740 and No. 1 245 384. The starting substances for these processes are peracyl glycosyl cyanates, which are obtained by adding O- alkylischarneas or S-alkylisothiureas into the corresponding peracylglycosyl isobiurets,

909847/1214

ORIGINAL

or Peracylglycosylisothiobiurete skip. These then supply 1-peracylglycosyl-4-alkoxy- (or -alkylthio-) - 1 _/ 2-dihydro-1 _/ 3 _/ 5-triazin-2-ones by condensation with orthoesters of aliphatic acids, which react with ammonia in alcohol or amines in alcohol are converted into l-glycosyl-5-azacytosines ₀ A disadvantage of this production process is that the yields of 1-glycosyl-5-azacytosines in the reaction of the ammonium or the amines with the l-peracylglycosyl in question 4-akoxy- (or alkylthio) -1,2-dihydro-1,3,5-triazin-2-ones fluctuate considerably and often only reach very low values. This fact is caused by the instability of the intermediates mentioned under the amination conditions used. In comparison to alkoxy derivatives, the instability of the alkylthio derivatives is even greater and their reaction rate slower, which together leads to deeper decomposition and a further reduction in the expansion of amination. It was found that 1-Peracylglykosy ± -4-alkoxy-1,2- -dihydro-1,3,5-triazine2-ones less stable than the free 1-glycosyl-4-alkoxy-1,2-dihydro-1 , 3,5-triazin-2-ones. The amination of the last-mentioned compounds proceeds very quickly and with only minor losses, so that they represent the most suitable intermediates for the preparation of l-glycosyl-5-azacytosines. The l-glycosyl-4-alkoxy-1,2-dihydro-1,3,5-triazin- -2-ones are easily produced both by alcoholysis of the corresponding peracyl derivatives and by alcoholysis of 1-peracylglycosyl- -4-alkylthio- 1,2-dihydro-1,3,5-triazin-2-ones accessible. This is because the alcoholysis of the last-mentioned compounds also results in a substitution of the alkylthio group by an alkoxy group.

9Q9847 / 12U

The subject invention of a production process for l-glycosyl-5-azacytosines of the general formula I consists in the reaction of compounds of the general formula II

II.

in which R has the same meaning as in formula I,

R is a peracylglycosyl with an acyl of 2-10 carbon atoms and X is an alkoxy or alkylthio group having 1 to 4 carbon atoms is, with an alkali metal alcoholate having 1-5 carbon atoms in an alkanol having 1-6 carbon atoms, with Advantage in methanoi, with formation of 1-glycosyl-4-alkoxy- -1,2-dihydro-1,3,5-triazin-2-ones of the general formula III

OR *

III.

1 2
in which R and R have the same meaning as in the formula I and R is an alkyl with 1-4 carbon atoms and these compounds are then further in an alkanol with 1-6 carbon atoms, advantageously in methanol, with compounds of the general formula IV

R ³ - NH - R ⁴ IV.

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• in which R and R have the same meaning as in formula I, brought to reaction.

The implementation of the alkoxy or alkylthiotriazinones of the general Formula II in l-glycosyl-5-azacytosine of the general formula I. is advantageously carried out without isolating the alkoxytriazinones of the general formula III.

The reaction of the compounds of general formula II with the corresponding alkali metal alcoholates is carried out in an alkanol made with 1 - 6 carbon atoms, advantageously in methanol and this at room temperature. In the case of the alkylthio derivatives, it is necessary to obtain an optimal yield 1 ^ 2 mol of the To use alcoholate on 1 mole of the starting compound.

The reaction of the Alkoxytriazinoae of the general formula III with a compound of the general formula IV is carried out at room temperature in an alkanol with 1-6 carbon atoms, with advantage in methanol.

The conversion of the compounds of general formula I into the corresponding l-glycosyl-5-azacytesines of general formula I. can be carried out in one stage by the simultaneous action of an alkali metal alcoholate and a compound of the general formula IV be made in alcohol.

The following implementation examples are for illustration purposes only of the process in question, but in no way to limit the manufacturing process used.

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5-azacytidine
(l-ß ~ D-ribofuranosyl-4-amino-1 _/ 2-dihydro-l _i 3 _/ 5-triazin-2-one)

example 1

A mixture of 0.5875 g of 1- (2,3,5-tri-O-benzoyl-β-D-ribofuranosyl) -4-methylthio-1,2-dihydro-1,3,5-triazin-2-one , from 5 ml of absolute methanol and 1.2 ml of a 1 N sodium methoxide solution

t in methanol is stirred at room temperature in a flask with attached potassium hydroxide drying tube until the starting substance dissolves. The solution remains at room temperature for a further 45 minutes and is then decationized on a column of 10 ml of a weakly acidic cation exchanger (H), which has been washed through with water and methanol before use. The methanolic eluate from the column (60 ml) is evaporated from a bath in vacuo at 30.degree. The evaporation residue is dissolved in 20 ml of methanol and again evaporated. The obtained crude crystalline 1-ß-D-Ribofuranosyl- -4-methoxyl-1 _/ 2-dihydro-1,3,5-triazin-2-one is without further purification in 4 ml of a 10/6 solution of dry ammonia in absolute Dissolved methanol. The solution remains in a sealed flask at room temperature for 30 minutes (the product separates out after 5 minutes) and then in the refrigerator for a further 12 hours at -10 ° C. The deposited 5-azacytidine is filtered off with suction, washed through with methanol and dried in vacuo. 0.216 g is obtained, d _o i. 88.6 % of the product; M.p. 232-234 C (decomposition) ₀

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Example 2

A mixture of 0.5875 g of 1- (2,3 _r 5-tri-0-benzoyl ~ ß-D-ribofuranosyl) -4-methylthio ~ 1,2-dihydro-1,3,5-triazin-2 ~ one from 4 ml of a 10% solution of dry ammonia into methanol and 1.2 ml of a 1 N solution of sodium methoxide in methanol in a flask with added potassium hydroxide drying tube until dissolution of the starting substance (3 minutes) umgaiJhrt ₀ the solution is still a further 45 Leave for minutes at room temperature (the crystalline product begins to separate out after 30 minutes) and then placed in the refrigerator for a further 12 hours at -10 C. The product is filtered off with suction, washed through with methanol and dried in vacuo. One wins

.162 g,
Settlement),

0.162 g, ie 66.4 % , of 5-azacytidine; M.p. 232 - 234 ° C (decomposition

Example 3

A mixture of 0.5715 g of 1- (2,3,5-tri-0-benzoyl-fi-0-ribofuranosyl) -4-methoxyl-1,2-dihydro-1,3,5-triazin-2-one , from 5 ml of absolute methanol and 1 ml of a 1 N sodium methoxide solution in methanol is stirred in a flask with attached potassium hydroxide drying tube until the starting substance dissolves (5 minutes). The solution is left at room temperature for a further hour and then processed in the same way as was described in Example 1. 0.219 g, ie 89.7 % , of 5-azacytidine are obtained; Mp. 232-234 ⁰ C (decomposition).

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Example 4

A mixture of 0.5715 g of 1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl) -4-methoxy-1,2-dihydro-1,3,5-triazin-2-one , from 4 ml of a 10 ^ strength solution of dry ammonia in methanol and 1 ml of a 1 N sodium methoxide solution in methanol is stirred in a flask with attached potassium hydroxide drying tube at room temperature until the starting substance dissolves (5 minutes). The mixture is then processed in the same way as in Example 2. 0.174 g, ie 71.3 % , of 5-azacytidine are obtained; M.p. 232-234 ° C (decomposition).

5-aza-2 ^/ '-deoxycitidine (1- (2-deoxy-ß -D-ribofuranosyl) -4-amino-1,2-dihydro-1 _/ 3 _f 5-

triazin-2-one)
Example 5

A mixture of 0.4956 g of l- (3,5-di-0-p-toluyl-2-deoxy-ß-D- -ribofuranosyl) -4-methylthio-1,2-dihydro-1,3,5-triazin-2-one; from 10 ml of absolute methanol and 1.2 ml of a 1 N sodium methoxide solution in methanol is placed in a flask with magnetic stirring at room temperature Leave the potassium hydroxide drying tube in place for 2 hours and 45 minutes (the starting substance goes into solution after 2 hours) «, The solution is then decationized on a column of 10 ml of a weakly acidic cation exchanger (H) that was previously mixed with was washed through with aqueous methanol. The methanolic eluate from the column (60 ml) is evaporated from a bath in vacuo at 30.degree. C., the residue is dissolved in 10 ml of methanol and evaporated again. The obtained raw syrupy 1- (2-deoxy-J3 ribofuranosyl) - -4-methoxy-1,2-dihydro-1,3,5-triazin-2-one is in 2 ml of a

909847 / 12U - ⁹ -

Dissolve a 10% solution of dry ammonia in methanol and leave the solution in a sealed flask at room temperature for 30 minutes (the crystalline product begins to separate after 15 minutes) and then put it in the refrigerator for a further 12 hours at -10 ° C . The product is filtered off with suction, washed through with methanol and dried in vacuo. 0.180 g, ie 79% , of 5-aza-2-deoxyoytidine are obtained; M.p. 196-198 C (with subsequent solidification).

Example 6

A mixture of 0.4956 g of 1- (3,5-di-0-p-toluyl-2-deoxy = β-D-ribofuranosyl) -4-methylthio-1,2-dihydro-1,3,5-triasin -2-one, from 4 ml of a 10 JSigen solution of dry ammonia in methanol and 1.2 ml of a 1 N sodium alcoholate solution in methanol is stirred magnetically at room temperature in a flask with attached potassium hydroxide drying tube until the starting substance dissolves (45 minutes). The solution remains for one hour at room temperature (during this time the crystalline product separates out) and is then placed in the refrigerator for a further 12 hours at -10 ° C. The product is filtered off with suction, washed through with methanol and dried in vacuo. 0.132 g, ie 58 %, of 5-aza-2-deoxyoytidine are obtained; M.p. 196-199 C (with subsequent solidification).

Example 7

A mixture of 0.4796 g of 1- (3,5-di-0-p-Teluyl-2-deoxy-ß-D- -ribofuranosyl) -4-methoxy-1,2-dihydro-1,3,5-triazin-2-one, from

- 10 -

900847 / U j 4

10 ml of absolute methanol and 1 ml of a 1 N sodium methoxide solution in methanol are stirred magnetically for 4 hours at room temperature in a flask with attached potassium hydroxide drying device (the starting substance dissolves after 2.5 hours). Further processing is carried out in the same way as was indicated in Example 5. 0.176 g, that is to say 77.2% , of 5-aza-2'-deoxycytidine are recovered; M.p. 197 - 198 ° C (with subsequent solidification),

Example 8

A mixture of 0.4796 1- (3,5-di-0-p-toluyl-2-deoxy-ß-D-ribofuranosyl) -4-methoxy-1,2-dihydro-1,2,5-triazine 2-one, made from 4 ml of a 10% solution of dry ammonia in methanol and 1.0 ml of a 1 N sodium methoxide solution in methanol, is stirred magnetically at room temperature in a flask with an attached potassium hydroxide drying tube until the starting substance has dissolved (45 minutes). The further processing is the same as that given in Example 6. 0.120 g is obtained, d _o i. 52.7 % of 5-aza-2'-deoxycytidine; M.p. 198-199 C (with subsequent solidification).

l-ß-D ~ ribofuranosyl-4-methoxy-1,2-dihydro-1,3,5-triazin-2-one Example 9

A mixture of 0.5875 g of 1-2,3,5-tri-O-benzoyl-ß-D-ribofuranosyl) -4-methylthio-1,2-dihydro-1,3,5-triazin-2-one, from 5 ml of absolute methanol and 1.2 ml of a 1 N sodium methoxide solution in methanol is added to a flask at room temperature

9 0 9 ü '- 7/12 I h

attached potassium hydroid drying tube until the starting substance dissolves (5 minutes). The solution is then left at room temperature for a further 40 minutes and then processed in the manner described in Example 1. The obtained crude crystalline product is recrystallized from absolute methanol. 0.210 g, or 81 % , of 1-A-D-ribofuranosyl-4-methoxy-1,2-dihydro-1,3,5-triazin-2-one are obtained; M.p. 177-179 C.

1-ß-D-ribofuranosyl-4-methylamino-l, 2-dihydro-1, 2, 5-triazin-2-one Example 10

A mixture of 0.259 g of 1-ß-D-ribofuranosyl-4-methoxy-1,2-dihydro-1, 3,5-triazin-2-one and 2 ml of a 7% solution of dry methylamine in absolute methanol is used Stirred at room temperature for 5 minutes in a flask with attached potassium hydroxide drying tube. Then the mixture is left after 10 minutes at room temperature and finally for a further 15 minutes at -10 C. The reaction product is filtered off with suction, washed through with ice-cold methanol and dried in vacuo. 0.210 g is recovered, di _f 81 4% l-.beta.-D-ribofuranosyl-4-methylamino-l, 2-dihydro-1, SfS-triazin ^ -one; Mp ₀ 148-150 C.

1-β-D-ribofuranosyl-4-dimethylamine o-1 _ff 2-dihydr o-i, 3,5-triazin--2-one

Example 11

A mixture of 0.259 g l-.beta.-D-ribofuranosyl-4-methoxy-l, 2-dihydro-l, 3 _i 5-triazin-2-one and dry from 2 mL of a 7 $ solution of dimethylamine in absolute methanol at room temperature Stirred for 5 minutes in a flask with an attached potassium hydroxide drying tube. The solution remains at this for a further 15 minutes

90984 7/1 2U

- 12 -

Room temperature and is then evaporated from a bath ^{in vacuo at 3O 0 C.} The residue is codistilled with 5 ml of absolute methanol and the syrupy product is dissolved in 1 ml of absolute ethyl alcohol. The frozen solution is then made to crystallize by friction and the mixture is left in the refrigerator (-10 C) overnight. The product is filtered off with suction, washed through with ethyl alcohol and dried in vacuo. 0.216 g, ie 79.4 % , of 1-β-D-ribofuranosyl-4-dimethylamino-1,2-dihydro-1,3,5-triazin-2-one are obtained; Smpo 128-130 ⁰ C ₀

- 13 -

9098A7 / 12U

Claims (3)

PATENT CLAIM: Process for the preparation of l-glycosyl-5-azacytosinefi der general formula I. I. 1 2 in which R is a glycosyl, R is hydrogen or an alkyl with 3 4 1-4 carbon atoms and R and R, identical or different, Hydrogen atoms, alkyls with 1-4 carbon atoms or Aralkyls with 7-10 carbon atoms, characterized in that a compound of the general formula II Ho 2 pp in which R has the same meaning as in formula I, R 9098A7 / 12U a peracylglycosyl with an acyl with 2-10 carbon atoms and X is an alkoxy or alkylthio group with 1-4 carbon atoms, with an alkali metal alcoholate with 1-6 carbon atoms in an alkanol with 1-6 carbon atoms, advantageously in methanol, and which resulting l-glycosyl-4-alkoxy-1,2-dihydro = 1,3j5-triazin-2-one of the general formula III ^ΙΠ 12th
in which R and R have the same meaning as in formula I and R is an alkyl with 1-6 carbon atoms, in an alkanol with 1-6 carbon atoms, advantageously in methanol, with a compound of the general formula IV R ³ - NH - R ⁴ IV 3 4 in which R and R have the same meaning as in formula I, are reacted. PATENT ADVOCATE 7/1214