DE2456547A1 - N HIGH 4 -SUBSTITUTED 2,2'-CYCLOCYTIDINE COMPOUNDS - Google Patents

Description

"N-substituted 2,2'-cyclocytidine compounds"

Priorities: November 29, 1973, Japan, No. 133 084/1973

■ December 4, 1973, Japan, No. 134 877/1973

- December 6, 1973, Japan, no.135 755/1973

It is known that 2,2'-cyclocytidine is used to combat leukemia. This compound has few side effects; See Y. Sakai et al., Japan J. Clin. oneol., Vol. 6 (1972), p. 57. Some cyclocytidine derivatives, namely N -substituted - ^, 2 · -cyclocytidines, are said to have at least the same effect against leukemia as 2,2'-cyclocytidine; see Proceedings of the Japanese Cancer Association, The 32nd Annual Meeting (1973), p 395. Furthermore, numerous 2,2 · cyclocytidine derivatives are known. For example, in GB-PS 1,325,798 2,2'-anhydro-ara-cytidine derivatives with a 5'-0-acyl group and a 4-imino salt group are described, which are said to be viral agents and anticancer agents and immunosuppressants. In JA-OS 16 481/1972 there are various 2'-0-, 2 ', 3'-di-O-, 2', 5'-di-O- and 3 ^f , 5'-di-0-esters described the excellent immunosuppressants, anticancer drugs and MLttei for the treatment of,

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Represent leukemia. Finally, US Pat. No. 3,709,874 describes i-β-D-arabinofuranosylcytosine derivatives which are viral agents and cytostatics are.

The invention is based on the object of providing new N -substituted, To create 2'-cyclocytidine compounds useful in the treatment of leukemia. This object is achieved by the invention solved.

The invention thus relates to that characterized in the claims Object.

The compounds of the general formula I include compounds of the general formulas Ia, Ib and Ic

HOH ₂ C

(Ia)

N - Y

- Y

(Ic)

N - Y

RC-OH ₂ C ^ 0 ^ ₀

(Ib)

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- 3 Y iind. R have the meaning given in claim 1.

Specific examples of the alkyl radicals R are the methyl, Ethyl, n-propyl, isopropyl, η-butyl, sec-butyl, tert-butyl, n-octyl, pivalyl, caprylic, stearyl and arachinyl groups. Unbranched or branched alkyl radicals having 1 to 17 carbon atoms are preferred.

Specific examples of aryl radicals R are the phenyl, 2,6> -dimethy! Phenyl, 2,4,6-trimethylphenyl-, p-methoxyphenyl-, 3,4,5-trimethoxyphenyl-, 2,4,6-trimethoxyphenyl-, p-chlorophenyl-, p-fluorophenyl-, p-nitrophenyl-, 2,3-xylyl-, 'isobut ^ rlphenyl-, p-tolyl and α- or ß-naphthyl group.

Specific examples of aralkyl radicals R are the benzyl, p-methylbenzyl-, p-ethylbenzyl-, p-isopropylbenzyl-, ß-naphthylmethyl-, 1-methylnaphthylmethyl, p-fluorobenzyl and o-chlorobenzyl group.

Specific examples of polycyclic hydrocarbon radicals R are the adamantyl-, (3-carboxymethyladamanten-1-yl) -methyl-, Norbornyl, · norbornenyl and 3,5,7-trimethyladamantylmethyl groups. ■ ·. ·

Specific examples of cycloalkyl radicals R are the cyclobutyl _T , cyclopentyl, cyclohexyl, 4-methylcyelohexyl, cyclooctyl and cyclodecanyl groups.

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The compounds of the general formulas Ia, Ib and Ic can be prepared in a manner known per se by acylating the corresponding Hydroxy compounds with an acylating agent, for example an acid anhydride of the general formula II or a Acid halide of the general formula III;

(RCO) ₂ O or RCOX (II) (III)

are prepared, in which R has the meaning given in claim 1 and X represents a chlorine, bromine or iodine atom; see Synthetic Procedures in Nucleic Acid Chemistry, Vol. 1, Pages 383 to 387 and pages 439 to 440.

The 3'-0-acyl-N-substituted-2,2'-cyclocytidines of the general formula Ia can be prepared by reacting a 5 ^f -0-trityl-N-substituted-2,2 ^f -cyclocytidine of the general formula IVa

N-Y

(IVa)

in which Y has the meaning given in claim 1, with a Acylating agents of the general formula II or III and hydrolysis of the 3'-0-acylate formed for splitting off the trityl group in the 5 'position. The selective one Cleavage of the trityl group without affecting the 3'-O-acyl group can be carried out in a manner known per se

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See Synthetic Procedures in Nucleic Acid Chemistry, Vol. 1, pages 383 to 387. '

4th
The N -hydroxy compounds of the general formula IVa used according to the process can be prepared in a manner known per se from the corresponding N -hydroxy-2,2'-cyclocytidine, for example by reacting an N -hydroxy-2,2'-cyclocytidine with a trityl halide such as trityl chloride in a basic solvent such as pyridine; See Synthetic Procedures in Nucleic Acid Chemistry, Vol. 1, pages 383 to 387. A typical example of the preparation of an N-amino-2,2'-cyclocytidine is described in the preparation examples below. The 5'-O-trityl-N-substituted-2,2'-cyclocytidine used according to the process can be present as a free base or as a salt of an acid.

The acylation of the compound of general formula IVa can be carried out with an acylating agent of general formula II or III in a molar ratio of acylating agent to compound of general formula IVa of 1 to 3: T, preferably 1 to 1.5: 1, in a solvent such as an amine, for example pyridine, dimethylamine or diethylamine, or a non-basic polar solvent such as dimethylacetamide, dimethylformamide or hexamethylphosphoric triamide, at temperatures from room temperature (about O to 30 ⁰ C) to 6O ⁰ C, preferably 20 to 60 ⁰ C is performed. The reaction time depends on the reaction temperature. Generally, about 24 to 72 hours are required. The course of the acylation can be followed, for example, by thin-layer chromatography *

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The 5'-O-trityl-3'-O- prepared in the above manner

acyl-N -substituted, 2'-cyclocytidine can first be isolated from the reaction mixture before the hydrolysis is carried out. The hydrolysis can also be carried out immediately after the solvent has been separated off, for example by distillation, from the reaction mixture and addition of an acid such as formic acid or acetic acid. The hydrolysis for splitting off the trityl group from the 5'-0-trityl-3'-0-acylate obtained can be carried out with an acid such as formic acid or acetic acid in an amount of about 20 to 50 ml, preferably 30 to 40 ml, per mmol 5'-O-trityl-3'-O-acylate be carried out at temperatures of about 20 to 100 ⁰ C for about 10 to 180 minutes.

The product is isolated from the reaction mixture by customary methods, for example by adding an organic solvent, which is a non-solvent to the product, such as acetone, diethyl ether, petroleum ether, benzene or Xylene. This causes the product to precipitate. The reaction mixture can also be evaporated under reduced pressure and the Residue from a solvent or solvent mixture, for example a lower aliphatic alcohol such as methanol or ethanol, or a lower alkyl ester of an aliphatic carboxylic acid such as ethyl acetate, are recrystallized. the. The aforementioned processes can also be used in combination with chromatography on silica gel, aluminum oxide, cellulose or an ion exchange resin.

The compounds of general formula Ib can be obtained by acylation of an N ⁺ -substituted-2,2'-cyclocytidine of the general

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nen formula IVb

N-Y

(IVb)

HOH ₉ C

² * 0

in which Y has the meaning given in claim 1, with an acylating agent of the general formula II or III will. In general, this acylation gives a mixture of the compounds It) and Ic. This mixture can by fractional recrystallization or chromatography in the components are separated.

The compounds of the general formula IVb used according to the process can be prepared from the corresponding compound in which Y represents a hydroxyl group, or, by the method described in the preparation examples, from the compounds in which Y represents an amino group. The starting compound can be in the form of a base or a salt of an acid. To increase the selectivity of the formation of the 5 ^! Acylate, the starting compound is preferably used in the form of a salt of an acid, for example as hydrochloride, sulfate or acetate. For the selective preparation of the 5'-0-acylates, the acylation can also be carried out in a non-basic polar solvent, such as dimethylacetamide, dimethylacetamide

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thylformamide or hexamethylphosphoric triamide, under anhydrous conditions. In this way, a mixture is * obtained that predominantly a 5'-O-acylate and only 2 to 3 percent of 3 ^1, 5'-di-O-acylate comprising the general formula Ic. The molar ratio of acylating agent of the general formula II or III to the compound of general formula IVb is about 1 to 3: 1, preferably 1 to 1.5 i 1. The reaction temperature is about 0 to 8O ⁰ C, preferably 20 to 60 ⁰ C. The reaction time is about 24 to 72 hours. The course of the acylation can be followed in a manner known per se, for example by thin-layer chromatography.

For the most selective preparation of the 3 ¹ , 5 '~ di-O-acylates, the acylation can be carried out under the usual conditions for acylating nucleosides in an amine, such as pyridine, dimethylamine or diethylamine, as a solvent; See Synthetic Procedures in Nucleic Acid Chemistry, Vol. 1, pages 383 to 387 and pages 439 to 440. In this way, a mixture of 3 ', 5'-di-0-acylate and 2 to 3 percent 5'- Prepare 0-acylate of the general formula Ib. The acylation can be carried out in the same way as described above for the 5'-0-acylation. The molar ratio of acylating agent of the general formula II or III to the compound of the general formula IVb is, however, about 2 to 10: 1, preferably 3 to 5: 1.

If the acylation of the N -substituted-2,2'-cyclocyti dine of the general formula IVb using a solution

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Solvents of a certain type carried out, so are predominantly the 5'-0-acylates or predominantly the 3 ', 5'-di-O-acylates obtain.

The products, i.e. the 5'-0-acylates or the 3 ', 5'-di-O-acylates, can be isolated from the reaction mixture in the same way as was described above for the isolation of the 3'-0-acylates is described.

The 3 ', 5'-di-0-acylates of the general formula Ic can also be prepared by acylating the corresponding monoacylate, that is to say the 3'-O-acylate of the general formula. Ia or the 5 ^t -0-acylate of the general formula Ib. getting produced. The acylation of the monoacylates to prepare the 3 '>5'-di-0-acylates is generally carried out in the same way as was described for the acylation of the 5 ^f -0-trityl compounds of the general formula IVa. The acylation can thus be carried out in an amine or a non-basic polar solvent of the type described above with a molar ratio of acylating agent of the general formula II or III to the monoacylate of about 1 to 10: 1, preferably 2 to 3: 1, at temperatures of about 0 · to 80 ^° C., preferably 20 to 60 ^° C., for about 24 to 72 hours. The course of implementation can! can be followed by thin-layer chromatography. The isolation of the 3 ', 5'-di-0-acylate can be carried out in the same way as was described above for the isolation of the 3'-0-acylate of the general formula Ia. In this way, 3 ', 5'-di-0-acylates with different acyl radicals in the 3 ^r - and 5' position from a monoacylate using an acylic

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be produced which differs from the acyl group of the monoacylate.

In the process for the preparation of the compounds of the invention, the starting compounds used for the acylation can be used in Can be used in the form of the free base or in the form of a salt of an acid. The salts can differ from inorganic acids, such as hydrochloric acid, hydrobromic acid, hydriodic acid or Sulfuric acid, or organic acids such as formic acid, acetic acid, maleic acid, fumaric acid, succinic acid, oxalic acid, Tartaric acid, citric acid, lactic acid or methanesulfonic acid, derive.

Any acyl halides can be used as acylating agents according to the invention or acid anhydrides of the general formula II or III for 3'-O-acylation, 5'-O-acylation or 3 ', 5'-di-O-acylation can be used. For 5'-O-acylation, preference is given to the acid halides of the general formula 'III are used because better yields of the corresponding 5'-0 ~ acylates or 3 ', 5'-di-0-acylates are obtained.

Most of the acyl halides of the general formula III used in the process according to the invention are known or are known can be obtained from the corresponding carboxylic acids of the general formula RCOOH, in which R has the above meaning, by reacting the carboxylic acid with a halogenating agent such as thionyl chloride, phosphorus tribromide or phosphorus trichloride, produce. The acyl iodides can be derived from the corresponding Acyl chlorides by reacting with an alkali metal iodide,

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preferably lithium iodide, in a solvent such as a ketone, "for example acetone or methyl ethyl ketone, or by Reacting with a quaternary ammonium iodide, such as tetraethylammonium j ο did, in a solvent like an ether, both

or tetrahydrofuran, for example ethylene glycol dimethyl ether, di-n-butyl ether / ' produce. The acid anhydrides of the general formula II are also known or can be obtained in a manner known per se produce.

Typical examples of acyl radicals of the general formula R-C-in the compounds of the invention are the acetyl, n-propionyl, n-butyryl, isobutyryl, caproyl, Palmitoyl, stearoyl, p-anisoyl, berizoyl, p-chlorobenzoyl, p-nitrobenzoyl, phenylacetyl, p-toluyl, o-toluyl, 3,4,5-trimethoxybenzoyl-, adamantan-1-yl-carbonyl-, 5-norbornenyl-2-carbonyl-, . Cyclobutylcarbonyl, trimethylacetyl-CPivaloyl), 2,6-dimethylbenzoyl and 2,4,6-trimethylbenzoyl groups.

Accordingly, the acylating agent used may be an acyl halide or an acid anhydride similar to the foregoing Corresponds to acyl residues. .

The compounds of the general formula I can be in the form of the free base or in the form of a salt of an acid. The salts are preferred because of their stability. The salts are prepared in a manner known per se from the free bases. For example, the hydrochlorides of the compounds of general formula I by reacting the '' free base with a 5 weight percent solution of chlorine

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hydrogen in a mixture of methanol and ethyl acetate. The salts can also be converted into other salts in a manner known per se, for example by treatment with an ion exchanger.

Specific examples of the 3'-0-, 5'-O and 3 ', 5'-di-0-acyl-N-substituted-2,2 ^f -cyclocytidine of the invention are given below:

, v 4

(I) 3'-0 "ACyI-N -hydroxv-2,2 ^> -cvclocytidine derivatives

(1) 3'-0-acetyl-N -hydroxy-2,2'-cyclocytidine and the hydro

chloride, hydrobromide and hydroiodide.

(2) 3'-0-propionyl-N -hydroxy-2,2'-cyclocytidine and the hydro

chloride, hydrobromide and hydroiodide.

(3) 3'-0-butyryl-N -hydroxy-2,2'-cyclocytidine and the hydro

chloride, hydrobromide and hydroiodide. ^%

(4) 3'-0-isobutyryl-N -hydroxy-2,2'-cyclocytidine and the hydro

chloride, hydrobromide and hydroiodide.

(5) 3'-0-Caproyl-N -hydroxy-2,2 ^f -cyclocytidine and the hydro

chloride, hydrobromide and hydroiodide.

(6) 3'-O-palmitoyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, Hydrobromide and hydroiodide.

(7) 3'-0-stearoyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, Hydrobromide and hydroiodide.

(8) 3'-0-p-anisoyl-N -hydroxy-2,2 ^! -cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(9) 3'-O-Benzoyl-N -hydroxy-2,2'-cyclocytidine and the hydro

chloride, hydrobromide and hydroiodide.

(10) 3'-0-p-chlorobenzoyl-N -hydroxy-2,2'-cyclocytidine and that

Hydrochloride, hydrobromide and hydroiodide.

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(11) 3'-0-p -nitrobenzoyl-N -hydroxy-2,2'-cyclocytidine and that

Hydrochloride, hydrobromide and hydroiodide.

(12) 3'-O-phenylacetyl-N -hydroxy-2,2'-cyclocytidine and that Hydrochloride, hydrobromide and hydroiodide.

(13) 3'-O-p-toluyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, Hydrobromide and hydroiodide.

(14) 3 »-o- (3,4,5-trimethoxybenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(15) 3'-O- (I-Adamantoyl) -N -hydroxy-2,2'-cyclocytidine and that Hydrochloride, hydrobromide and hydroiodide.

(16) 3'-0- (5-Norbornenyl-2-carbonyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(17) 3 · -O-cyclobutylcarbonyl-N -hydroxy-2,2 · -cyclocytidine and the hydrochloride, hydrobromide and hydroiodide. '

(18) 3'-0-pivaloyl-N-hydroxy-2,2 ^Zt · -cyclocytidin

and the hydrochloride, hydrobromide and hydroiodide.

(19) 3 ^f -0- (2,6-Dimethylbenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(20) 3'-0- (2,4,6-trimethylbenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(II) 3'-0-Acvl-N -amino-2,2'-cyclocytidine derivatives (21.) 3'-0-acetyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide .

(22) 3'-0-propionyl-N ^{Z |} -amino-2,2-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(23) 3'-O-butyryl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

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(24) 3'-0-isobutyryl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(25) 3'-0-caproyl-N -amino-2,2 ^! -cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(26) 3'-O-palmitoyl-N -amino ~ 2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(27) 3'-O-stearoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(28) 3'-0-p-anisoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(29) 3'-O-Benzoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and Hydrojodi'd.

(30) 3'-0-p -chlorobenzoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(31) 3'-Op-nitrobenzoyl-N -amino-2,2'-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(32) 3'-O-phenylacetyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(33) 3'-o-p -toluyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(34) 3'-0- (3,4,5-trimethoxybenzoyl) -N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(35) 3'-0- (1-Adamantoyl) -N ⁴ -amino-2,2-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(36) 3'-0- (5-norbornenyl-2-carbonyl) -N -amino-2,2'-cyclo-

. cytidine and the hydrochloride, hydrobromide and hydroiodide.

(37) 3 ^f -O-cyclobutylcarbonyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

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(38) 3'-O-pivaloyl-N-amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(39) 3'-O- (2,6-dimethylbenzoyl) -N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(40) 3'-0- (2,4,6-trimethylbenzoyl) -N -amino-2,2 »-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(III) 5'-0-acyl-N -hydroxy-2,2'-cyclocytidine derivatives

(41) 5'-O-acetyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(42) 5'-0-propionyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(43) 5 "-O-butyryl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(44) 5ι-O-isobutyryl-N-hydroxy-2,2'-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(45) 5'-O-caproyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroodide.

(46) 5'-O-palmitoyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(47) 5'-O-stearoyl-.N -hydroxy-2,2'-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(48) 5'-O- p -anisoyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(49) 5'-O-Benzoyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(50) 5'-O-p -chlorobenzoyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

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(51) 5'-0-p -nitrobenzoyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(52) 5 ¹ -O-phenylacetyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(53) ^{5'-0-p-toluyl-N-hydroxy-2,2'-Zf} cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(54) 51-0- (3,4,5-trimethoxybenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(55) 5 ^f -0- (i-Adamantoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(56) 5'-0- (5-Norbornenyl-2-carbonyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(57) 5'-0-cyclobutylcarbonyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(58) 5'-O-pivaloyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(59) 5'-0- (2,6-Dimethylbenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(60) 5'-0- (2,4,6-trimethylbenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(IV) 5'-0-Acvl-N -amino-2 _t 2'-cyclocytidine derivatives

(61) 5'-O-acetyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(62) 5'-0-propionyl-N ^Zf amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(63) 5 ^f -O-Butyryl-N-amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

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(64) 5'-0-isobutyryl-N -amino-2,2-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(65) 5'-O-caproyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(66) ^ '- O-Palmitoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(67) 5'-O -stearoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(68) 5'-O- p -anisoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(69) 5'-O-Benzoyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(70) 5'-Op-chlorobenzoyl-N -amino-2,2'-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(71) 5'-Op-nitrobenzoyl-N -amino-2,2'-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(72) 5'-0-phenylacetyl-M -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(73) 5'-O-p -toluyl-N -amino-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(74) 51_o_ (3,4,5-trimethoxybenzoyl) -N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(75) 5'-0- (i-Adamantoyl) -N ⁴ -amino-2,2'-cyclocytidine
and the hydrochloride, hydrobromide and hydroiodide.

(76) 5'-0- (5-Norbornenyl-2-carbonyl) -N -amino-2,2 · -cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(77) 5'-O-cyclobutylcarbonyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

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(78) 5'-0-pivaloyl-N -amino-2,2-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(79) 5'-O- (2,6-dimethylbenzoyl) -N -amino-2,2'-cyclocytidine • - and the hydrochloride, hydrobromide and hydroiodide.

(80) 5'-0- (2,4,6-trimethylbenzoyl) -N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(V) 3 ', 5 ' -Di-0-acyl-N -hydroxy-2,2'-cyclocytidine derivatives

(81) 3 ', 5'-di-0-acetyl-N ⁴ -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(82) 3 ^1, 5'-di-0-propionyl-N-hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(83) 3 ', 5'-di-0-butyryl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(84) 3 ', 5'-di-O-isobutyryl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(85) 3 ', 5'-di-0-caproyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(86) 3 ', 5'-di-0-palmitoyl-N ^ -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(87) 3 ', 5'-di-0-stearoyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(88) 3 ^1, 5'-di-0-p-anisoyl-N-hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(89) 3 ', 5'-di-0-benzoyl-N ^{Zt-hydroxy-2,2'-cyclocytidine} and the hydrochloride, hydrobromide and hydroiodide.

(90) 3 ', 5'-di-0-p-chlorobenzoyl-N-hydroxy-2,2 ^Zt "-cyclocytidin and the hydrochloride, hydrobromide and hydroiodide.

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(91) 3 ¹ , 5'-di-0-p-nitrobenzoyl-N -hydroxy-Z ^ '- cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(92) 3 ', 5'-Di-O-phenylacetyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(93) 3 ', 5'-Di- O- p -toluyl-N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(94) 3 ', 5'-di-0- (3,4,5-trimethoxybenzoyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(95) 3 ', 5'-di-0- (i-adamantoyl) -N -hydroxy-2,2'-cyclocytidine

'and the hydrochloride, hydrobromide and hydroiodide. -

(96) 3 ', 5'-Di-0- (5-norbornenyl-2-carbonyl) -N -hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(97) 3 ', 5'-di- O -cyclobutylcarbonyl-N -hydroxy-2,2-cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(98) 3 ', 5'-di-O-pivaloyl-N -hydroxy-2,2'-cyclocytidine

and the hydrochloride, hydrobromide and hydroodide.

(99) 3 ^1, 5'-di-0- (2,6-dimethylbenzoyl) -N-hydroxy-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(100) 3 ', 5'-di-0- (2,4,6-trimethylbenzoyl) ^{-N-hydroxy-2,2'-Zf} cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(VI) 3 \ 5 ^X -Di-0-acyl-N -amino-2,2'-cyclocytidine derivatives

(101) 3 '■, 5' -Di-0-acetyl-N ^ -amino-2,2 · -cyclocytidine

and the hydrochloride, hydrobromide and hydroiodide.

(102) 3 ', 5'-di-0-propionyl-N ^Zf amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

S09833 / 0S45

(103) 3 ', 5'-Di-0-butyryl-N ⁴ -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(104) 3 ', 5'-di-0-isobutyryl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(105) 3 ', 5'-Di-0-caproyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(106) 3 ', 5'-di-0-palmitoyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(107) 3 ', 5'-di-0-stearoyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(108) 3 ', 5'-di-0-p-anisoyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(109) 3 ', 5'-Di-0-benzoyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(110) 3 ', 5'-di-0-p -chlorobenzoyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(111) 3 ', 5'-Di-0-p -nitrobenzoyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(112) 3 ', 5'-Di-O-phenylacetyl-N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(113) 3 ¹ , 5'-di-0-p-toluyl-N -amino-2, 2 »-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(114) 3 ', 5'-Di- O- (3,4,5-trimethoxybenzoyl) -N -amino-2,2 · -cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(115) 3 ', 5'-Di- O- (1-adamantoyl) -N -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(116) 3 ', 5'-Di-O- (5-norbornenyl-2-carbonyl) -N ⁴ -amino-2,2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

509333/05 ^ 5

(117) 3 ¹ , S'-Di-O-cyclobutylcarbonyl - ^ - ainino ^, ^ '-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(118) 3 ', 5'-Di-O-pivaloyl-N ^ -amino-aa ¹ -cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

(119) 3 ', 5'-di-O- (2, 6-dimethylbenzoyl) -N ^Zt -amino-2,2' -cyclocytidin and the hydrochloride, hydrobromide and hydroiodide,

(120) 3 ·, 5 · -Di-0- (2,4,6-trimethylbenzoyl) -N -amino-2 _t 2'-cyclocytidine and the hydrochloride, hydrobromide and hydroiodide.

The compounds of general formula I can together with customary carriers and / or diluents and / or auxiliaries orally, intravenously, intraperitoneally, subcutaneously or administered rectally. Injectables can be made by dissolving a compound of the general formula I in distilled water, physiological saline solution or Ringer's solution getting produced. Compounds of the general formula I that are sparingly soluble in an aqueous medium can be converted into sodium carboxymethyl cellulose, Sodium alginate, polysorbate 80 or tragacanth can be suspended. For oral administration can the pharmaceuticals are packaged in the form of tablets, capsules, pills, powders, syrups, emulsions or suspensions will. Carriers for drugs for oral administration are, for example, lactic acid, cane sugar, starch, corn starch, Calcium carbonate, talc, magnesium stearate, sodium carboxymethyl cellulose, gum arabic, tragacanth and polysorbates 80 in question. Examples of carriers for suppositories are cocoa butter, glycerine and polyethylene glycol.

509833/0545

- 22- The daily dose can be 5 to 30 mg / kg

The compounds of general formula I and their salts are anticancer agents, in particular agents for combating leukemia. The determination of the cytostatic activity of the compounds of general formula I and known comparison compounds was made according to the method of Cancer Chemotherapy National

(Japan), Service Center, Cancer Chemother. Rep./Bd. 25 (1962), p. 1,

performed with the following tumors:

lymphatic leukemia 1210-L (L-1210), Nakahara-Fukuoka

and maleji__ sarcoma (NFS) and Ehrlich ascites carcinoma (EAC) in female / BDF ₁ mice with a body weight of 20 ± 2 g and 6 animals per group. In the case of solid tumors, that is

NFS, 20 to 40 mg of the tumor fragments are implanted subcutaneously in the interscapular region of the mouse. Ehrlich ascites tumors and the leukemia 1210 tumors are given to the mice

7 intraperitoneally in an amount of 1 × 10 cells or

5
1 χ 10 cells implanted.

The compound to be investigated is administered in a daily dose of 100 mg / kg intraperitoneally as a suspension in carboxymethyl cellulose administered once a day for 5 days 24 hours after the cells were transplanted. The evaluation at L-1210 is based on the mean survival time and is expressed as a percentage of the mean survival time of the control animals expressed. For NFS solid tumors, the tumor weight ratio (T / C percent) is determined on the 10th day after implantation and in the case of the EAC ascites tumors, the cell volume ratio (T / C percent) is measured on the 7th day.

509833/0545

The activity of typical compounds of the general formula I is given in Table I. It can be seen from the results that the compounds of the invention increase the mean life of test animals suffering from leukemia compared to 2,2'-cyclocytidine-, 1-ß-D-arabinofuranosylcytosine (Ara-C), N ⁴ -hydroxy-2, 2 «-cyclocytidine and 5-0-acetyl-2,2 ^f cyclocytidine (GB-PS 1,325,798) and 3 ', 5'-di-0-acetyl-2.2 ^l cyclocytidine (US-PS 3,709,874) cause.

In Table I, the mean survival time ILS percent was calculated using the following equation:

/ Average over-] / Average over- '(lifetime of behan- J -1 times the j ^ un- group S days yhandelten group, days /

ILS96 100 χ

(Average survival time - untreated Group, days)

The tumor weight ratio T / C% in NFS is as follows Calculated equation:.

Φ / w _ -] oo χ · tumor weight of the treated group

Tumor weight of the untreated group

The cell volume ratio Τ / C % in EAC is calculated using the following equation:

_ 100 χ tumor volume of the treated group

Tumor volume of the untreated group

509833/0545

Table I.

cn ο co oo to

-OH -OH -NH,

-OH

-OH

-OH

-OH

-OH

-NH ₂

-OH

-OH

-OH

-OH

Acyl radical in the 3 »position

f> co-

CH-CO-CH ₃ CO-palmitoyl

CH ₃ CO-

Acyl residue in 'der

• 5 ^f position ■ ._

CH ₃ CO CH ₃ CO

Adamantoyi PC ₆ H ₄ (CH ₃ ) CO-

CH ₃ CO-

CH ₃ CO-

CH ₃ CO-

'' Tumor type "" • ■ ircs' ■ EAC L-1210 (T / C%) (T / C%) (ILSI) 40- 30th 120 50 30th 130 60 30th 90 20th 20th 170 40 50 140 30th 30th 130 30th 30th 130 40 20th 140 60 60, 60 20th 10 180 '40 30 ' 130 40 20th 130 50 30th 150 70 40 • 50

Table I - continued

-OH OH

control

.Acyl radical ija the 3'-position - ~

Stearoyl

Caproyl

2.2 ^! -Cyclocytidine ..

N ⁴ -hydroxy-2,2 · cyclocytidine

5'-O-acetyl-2,2 · cyclocytidine

Macaw - C.

S'.S'-O-di-O-acetyl-2,2'-cyclocytidine

Macaw - C

Tumor type

Acyl residue in the
5 'position - _: 'L-1210 ¹ : NFS · ^1st EAC (ILS%) (T / C.%) (T / C%) Stearoyl 180 20th 20th Caproyl 150 30th 30th (100 mg / kg) 120 50 40 (100 mg / kg) 80 50 50 (100 mg / kg) ' 100 50 50 (3.0 mg / kg) 60 50 20th 120 60 50 (100 mg / kg) 20th Signs Display

of -Toxi- of- Toxicity ity '"■

The invention is illustrated by the preparation examples and examples. Parts, percentages and proportions relate to weight, unless otherwise stated.

Preparation Example 1 '· Preparation of N -aminocytidine

2 g (8.2 mmol) of cytidine are dissolved in 25 ml of a 4 η aqueous solution of hydrazine hydrate (pH 6.0). The solution is ^{heated to 80 °} C. for 6 hours. The reaction mixture is then evaporated to dryness and the water contained in the residue is distilled off twice with ethanol as an azeotropic mixture. A small amount of chloroform and ethanol are then added to the residue. After standing for 15 to 18 hours, the precipitated crystals are filtered off and washed with a small amount of ethanol. The filtrate and the

will
Wash solution / combined, evaporated to dryness and the residue dissolved in 50 ml of water. The aqueous solution is treated with 20 ml of an ion exchange resin Dowex 1x2 (HCO-z type). The ion exchange resin is filtered off and washed with water. The filtrate and the washing solution are combined and evaporated to dryness. The very viscous residue is mixed with 20 ml of acetone in order to separate off excess hydrazine. The insoluble residue is then washed several times with acetone. The very viscous product that remains is dried over phosphorus pentoxide. Yield 1.7 g (80.1 percent of theory) of caramel-colored N-aminocytidine. UV absorption spectrum:

, 275 ny, ^ £ _x ¹¹ '269 ψ.

509833/0545

217, 28 2 mjLi
Paper chromatography: R _f = 0.25 (n-butanol: acetic acid: '

Water = 5: 2 ϊ 3 volume)

C ₉ H ₁₄ O ₅ N ₄ ; "C HN

. calc .: 41.86 5.46 21.70 found: 41.90 5.30 21.41.

Production example 2

Production of 3'-0-Acetvl-N -amino-2,2'-CVClOCVtJdJn-hydrochlo-

rid,

0.85 g of N -aminocytidine be added to a mixture of 3.78 ml of phosphorus oxychloride, 0.74 ml of water and 5.97 ml of ethyl acetate, und.das mixture is heated 30 minutes 6O ⁰ C. ■ then is added to a mixture of 1.89 ml of phosphorus oxychloride, 0.37 ^{ml ¥ as} "ser and 2.98 ml of ethyl acetate and reacted, the reaction mixture for a further 4 hours at 70 ⁰ C. Then the reaction mixture was evaporated and the residue while cooling with The aqueous solution obtained is poured onto a column filled with 60 ml of a cation resin exchanger DIAION SK-IBS in the acid form. After washing the column with water, the column is filled with 700 ml of a 0.1 molar pyridine-formic acid solution (pH 4.5) and the eluate discarded. The fractions obtained on eluting the column with 1 liter of a 0.3 molar pyridine-formic acid solution (pH 4.5) are combined and evaporated. A yellow, amorphous product is obtained. This product is mixed with water and evaporated. This process is repeated several times to separate residual pyridine. Then the 'residue is dissolved with water to a volume of 10 ml. The solution is adjusted to a pH of 3 with 1 η hydrochloric acid. The acidic solution is evaporated again, in a _C e-

5Q9833 / 0SA5

wrestle amount of a mixture of isopropanol, acetic acid and Dissolved water in a volume ratio of 5: 1: 2 and attached to a column filled with 70 g of silica gel for chromatographic purposes chromatographed. The column is filled with a mixture of isopropanol, acetic acid and water in a volume ratio 5: 1: 2 eluted. The product fraction is evaporated and a small amount of acetone is added to the residue. Then the mixture is left to stand at room temperature. 240 mg (22.9 percent of theory) of 3'-O-acetyl-N -amino-2,2'-cyclocytidine hydrochloride are obtained obtained with a mp of 180 ° C (decomp.). UV absorption spectrum: ^ jjjg £ 263, 273 (s), 285 (s) mu

267, 285 (s) mu

IR spectrum: V ^ max 2600, 3000, 1740 cm ". C ₁₁ H ₁₅ O ₅ N ₄ Cl, CHN

calc .: 41.45 4.74 17.58 found: 41.66 4.72 17.59.

Production example 3

Preparation of N-amino-2,2'-cyclocytidine hydrochloride

4 is in

1 g 3 * -0-acetyl-N-amino-2,2'-cyclocytidine hydrochloride / 6Q ml of methanol, treated with 1 ml concentrated hydrochloric acid and allowed to stand for 6 days at 37 ⁰ C. The reaction mixture is then evaporated and the residue is digested with 30 ml of acetone. The mixture is filtered and the filter residue is washed thoroughly with acetone. Yield 0.655 g (75.4 percent of theory) of N-amino-2,2-cyclocytidine hydrochloride.

UV absorption spectrum: A ^H 2 ° 263, 273 (s), 285 (s) mu

max '

¹ 267,

509333/0545

₃ O ₄ N ₄ Cl; 39 C. H 74 N ber. ': 38 , 07 4, 52 20.25 found: , 57 4, .20.60

example 1

520 mg (1 mmol) 5'-0-trityl-N-hydroxy-2,2'-cyclocytidine-hydrQ-chloride (prepared from N-hydroxy-2,2'-cyclocytidine and trityl chloride according to Synthetic Procedures in Nucleic Acid Chemistry, Vol. L, 'pages 383 Ms 387) are dissolved in 30 ml of dimethylacetamide, mixed with 281 mg (2 mmol) of benzoyl chloride and left to stand for 7-2 hours at room temperature. After completion of the reaction, the solvent is distilled off under reduced pressure: the residue with 30 ml 98prozentiger formic acid are added and allowed to stand for 10 minutes at 2O ⁰ C. The formic acid is then distilled off under reduced pressure. The residue is mixed with water and evaporated. This process (addition of water and evaporation) is repeated several times until the formic acid has been completely separated off. The residue obtained is washed thoroughly with diethyl ether in order to separate off trityl alcohol. The product, which is insoluble in diethyl ether, is dissolved in a small amount of methanol and the solution is chromatographed on a column of dimensions 5 × 60 cm filled with silica gel for chromatographic purposes. The column is eluted with a mixture of methyl ethyl ketone, acetone and water (volume ratio 65: 20: 15). The product fractions are combined and evaporated under reduced pressure. Yield 88 mg (23 percent of theory) of crystalline 3'-O-benzoyl-N-hydroxy-2,2-cyclocytidine hydrochloride. That

509833/0545

Analysis preparation is recrystallized from ethanol.

UV Absorption Spectrum: ^ ^CH 3 ^0H (^ _x 10 " ³ ), 255 mu (16.2).

Max .

^c 16 ^H 16 ° 6 ^N 3 ^C1j C. HN

calc .: 50.34 4.22 11.01

found: 50.50 4.13 11th, 21st

Example 2

565 mg (1 mmol) of 5'-O-trityl-N-hydroxy-2,2'-cyclocytidine hydrobromide are dissolved in 30 ml of anhydrous pyridine. The solution is added dropwise under ice cooling with 246 mg (2 mmol) of acetyl bromide and allowed to stand for 40 hours at 5 ⁰ C. When the reaction has ended, 5 ml of methanol are added to the reaction mixture and the mixture is left to stand at room temperature for 2 hours. The pyridine is then distilled off under reduced pressure. The residue is dissolved in 40 ml of 80 percent acetic acid and refluxed for 3 hours. After cooling, the solution is evaporated to dryness. The residue is washed thoroughly with diethyl ether to separate off the trityl alcohol. The product, which is insoluble in diethyl ether, is dissolved in a small amount of methanol and the solution is

4 game 1 chromatographed. The fractions with 3'-0-acetyl-N hydroxy-2,2'-cyclocytidine are combined and evaporated. The residue is treated with a 5 percent solution of hydrogen bromide added in methanol. The hydrobromide which has crystallized out is filtered off. Yield 131 mg (36 percent of theory).

UV absorption spectrum: 3 ^H 2 ° (<f χ 10 " ⁵ ), 253 mu (16.8)

max ^;

509833/0545

^C 1 1 H ₁₄ O ₆ N ₃ Br; 36 C. H 11 N ber .: 36 , 28 3.87 11 , 53 found: , 23 4.11 , 48

Example 3

520 mg (1 mmol) of 5'-O-trityl-N-hydroxy-2.2 ^l -cyclocytidine hydrochloride are dissolved in 30 ml of anhydrous pyridine, mixed with 204 mg (2 mmol) of acetic anhydride and left to stand for 24 hours at room temperature. After the reaction has ended, 5 ml of methanol are added to the reaction mixture

Left to stand at room temperature for 2 hours. Then it becomes pyridine distilled off under reduced pressure. The residue is dissolved in 40 ml of 80 percent acetic acid and 3 hours heated to reflux. After cooling, the mixture is evaporated to dryness under reduced pressure. The residue is dissolved in a small amount of ethanol and the solution is stirred into 200 ml of ice-cold diethyl ether. The resulting precipitate is filtered off, dried and in a a small amount of methanol dissolved. The solution is chromatographed according to Example 1. The product fractions are combined and evaporated. A 5 percent solution of hydrogen chloride in methanol is added to the residue. The hydrochloride which has crystallized out is filtered off. Yield 67 mg (21 percent of theory).

UV absorption spectrum ^ ^H 2 ° (£ χ 10 ~ ³ ), 253 mu (16.8)

Max '

C ₁₁ H ₁₄ O ₆ N ₃ Cl; C _HN · '·.

ber .: 41.33 4.41 13.14 found: '41.00 4.28 13.21.

S09833 / 0SA5

Example 4

519 mg (1 mmol) 5'-O-Trityl-N -amino - ^^ '- cyclocytidine-hydro-

4th
Chloride (prepared from N-amino-2.2 ^l -cyclocytidine and trityl chloride according to Synthetic Procedures in Nucleic Acid Chemistry, Vol. 1, pages 383 to 387) are suspended in 20 ml of ethylamine. The suspension is admixed with 204 mg (2 mmol) of acetic anhydride while cooling with ice and stirring, and the mixture is left to stand at room temperature for 24 hours. When the reaction has ended, 5 ml of methanol are added to the reaction mixture and the mixture is left to stand for 2 hours. The solvent is then distilled off under reduced pressure and the residue is washed thoroughly with diethyl ether to separate off the amine. The insoluble product in diethyl ether is treated with 98prozentiger formic acid and allowed to stand for 10 minutes at 20 ⁰ C. The formic acid is then distilled off under reduced pressure, water is added to the residue and the mixture is evaporated. This process (adding and distilling off water) is repeated several times until the formic acid has been completely separated off. The residue obtained is dissolved in ethanol and poured into 200 ml of ice-cold diethyl ether with stirring. The resulting precipitate is filtered off and the filter residue is dissolved in a mixture of isopropanol, acetic acid and water in a volume ratio of 5: 1: 2. The solution is applied to a column filled with 80 g of silica gel for chromatographic purposes. The product fractions obtained on eluting with a mixture of isopropanol, acetic acid and water in a volume ratio of 5: 1: 2 are combined and evaporated. The residue is recrystallized from a 5 percent solution of hydrogen chloride in methanol. Yield 54 mg (17 percent of theory)

509833 / OSAS

31-0-acetyl-N -amino-2,2 ¹ -cyclocytidine hydrochloride. UV absorption spectrum: ^ ^H 2 ° (£ χ TO " ³ ) 263, 273 (s),

Max

286 (s) mu (13.6, 11.5, 4.2)

^C 1 1 ^H 1 5O ₅ N ₄ Cl; 41 C. H 74 1 N 58 ber .: 41 , 45. 4, 72 1 7, 63. found: , 38 4, 7,

Example 5

320 mg (1 mmol) of 3'-O-acetyl-N-hydroxy-2,2'-cyclocytidine hydrochloride are suspended with stirring in 20 ml of anhydrous pyridine and 3 mmol of acetic anhydride are added dropwise over 30 minutes while stirring. After a while the reaction mixture becomes transparent. After löstündiger reaction, 30 ml of methanol are added and the reaction mixture is heated for 30 minutes at 50 ⁰ C and then evaporated under reduced pressure. The residue is dissolved in anhydrous ethanol and evaporated to separate off the remaining pyridine. This procedure (addition of methanol and evaporation) is repeated three times. The semi-solid residue is recrystallized from ethyl acetate containing small amounts of methanol. Yield 202 mg (62 percent of theory) of '3', 5'-di-O-acetyl-N -hydroxy-2, 2'-cyclocytidine. When using a mixture of a 5 percent solution of hydrogen chloride in methanol and ethyl acetate for recrystallization, the hydrochloride is obtained.

• ■■. "" ■ · ■■ · ■ '

The free base has the following properties:

Ch γπ-τ ^ UV absorption spectrum: ^ 3 (£ χ ΙΟ "" - '), 254 mu (16.2)

max Γ

SO9833 / 0S4S

Nuclear Magnetic Resonance Spectrum (δ): 5.41 ppm (d, 1H, C ₂ , -H), 2.12 ppm (d, 6H,

- methyl proton). C ₁₃ H ₁₅ O ₇ N ₃ ; CHN

"calc .: 48.00 4.65 12.92. * found: 47.76 4.37 13.21.

Example 6

382 mg (1 mmol) of 5'-0-benzoyl-N-hydroxy-2,2'-cyclocytidine hydrochloride are suspended in 20 ml of anhydrous diethylamine with stirring and 3 mmol of benzoic anhydride are added over the course of 30 minutes while stirring. After a while the reaction mixture becomes clear. After I6stündiger reaction 3 ml of methanol are added and the reaction mixture is heated for 30 minutes at 50 ⁰ C and then evaporated under reduced pressure. The residue is dissolved in anhydrous ethanol and evaporated to separate off residual diethylamine. This procedure (addition of ethanol and evaporation) is repeated three times. The semi-solid residue is recrystallized from ethyl acetate containing a little methanol. Yield 234 mg (52 percent of theory) of 3 ', 5'-di-O-benzoyl-N hydroxy-2,2 ^f -cyclocytidine. When using a mixture of a 5 percent solution of hydrogen chloride in methanol and ethyl acetate, the hydrochloride is obtained.

The free base has the following properties:

^ ^CH 3 ^0H (6 χ ΙΟ " ³ ), 254 αμ (18.4) max '

Nuclear Magnetic Resonance Spectrum (δ): 5.41 ppm (d, 1H, C ₂ , -H)., 7.76 ppm (d, 1OH,

aromatic ring proton). ■

509833/0545

O ₇ N ₃ ; 61 C. H 26th N 35 ber .: 61 , 47 4, 43 9, 21 found: , 28 4, 9,

• Example 7 *.

320 mg (1 mmol) of 3 ^f -0-acetyl-N-hydroxy-2,2'-eyclocytidine hydrochloride are suspended in 20 ml of anhydrous pyridine and 3 mmol of acetyl chloride are added dropwise over 30 minutes while stirring. After some time, the reaction mixture becomes clear. After 20 hours of reaction, 3 ml of methanol are added, and the reaction mixture is left to stand for 2 hours at room temperature and then, according to Example 5,

U is working. The 3 ', 5'-di-0-acetyl-N -hydroxy-2,2'-cyclocytidine is obtained. According to Example 5, the hydrochloride is produced. The product has the same properties as the product obtained in Example 5.

• Example 8

■ · 4
319 mg (1 mmol) of S'-O-acetyl-N ~ amino-2.2 ^l -cyclocytidine hydrochloride are suspended in 20 ml of anhydrous pyridine, slowly admixed with 1.02 g (10 mmol) of acetic anhydride and at room temperature for 24 hours ditched. When the reaction has ended, the solvent is distilled off under reduced pressure, water is added to the residue and the mixture is evaporated. This procedure (addition of water and evaporation) is repeated several times. The semi-solid residue is recrystallized from ethyl acetate containing a little methanol. Yield 195 mg (60 percent of theory) of 3 ^! , 5'-Di-O-acetyl-N ^ - amino-2,2'-CyClOCy ^ dXn. When using a mixture of

S09833 / 0S4S

the hydrochloride is obtained from a 5 percent solution of hydrogen chloride in methanol and ethyl acetate.

The free base has the following properties: UV absorption spectrum:

(£ x 10 " ³ ), 263, 273 (s), 285 mu (15.0, 11.8, 5.2) max '

^ I ²⁶⁷ >

^C 13 ^H 16 ° 6 ^N 4i ^CHN

calc .: 48.15 4.97 17.28 found: 48.12 4.98 17.30

Example 9

278 mg (1 mmol) of N- ^{hydroxy-2.2 l} -cyclocytidine hydrochloride dissolved in 25 ml of anhydrous dimethylacetamide, mixed with 281 mg (2 mmol) of benzoyl chloride and left to stand for 72 hours at room temperature. After the reaction has ended, the solvent is distilled off under reduced pressure and the residue is washed with a mixture of ethyl acetate and diethyl ether in a volume ratio of 1: 2. A small amount of the powder obtained is dissolved with ethanol and chromatographed on a column with dimensions of 5 x 60 cm filled with silica gel for chromatographic purposes The product fractions obtained are combined and evaporated under reduced pressure. Yield 160 mg (42 percent of theory) of crystalline 5 ^f -O-benzoyl-N -hydroxy-2,2'-cyclocytidine hydrochloride, the

is recrystallized again from ethanol.

S09833 / 0S4S

"- 37 -

The product has the following properties:

UV absorption spectrum: D CH ₃ OH ^ - ₁₀ -3 ^ ₂₅₅ " _u (13.5)

^rK max '

Nuclear Magnetic Resonance Spectrum (δ): 8.30 ppm, 8.09 ppm (d.d, 1H, Cg-H), 5.41 ppm

(d, 1H, C ₂ , -H), 7.75 ppm (s, 5H, aromatic ring proton).

^C 16 ^H 16 ^N ° 6 3 ^C1; CH _N

calc .: 50.34 4.22 11.01

found: 50.23 4.35 11.33.

"Example

'4

322 mg (1 mmol) of N. hydroxy-2.2 ^t -cyclocytidine hydrobromide are dissolved in 25 ml of anhydrous dimethylacetamide, 246 mg (2 mmol) of acetyl bromide are added dropwise and the mixture is left to stand for 80 hours at room temperature. After the reaction has ended, the solvent is distilled off under reduced pressure and the residue is washed several times with a mixture of acetone and diethyl ether in a volume ratio of 1: 4. The product, which is insoluble in the mixture of acetone and diethyl ether, is recrystallized from a small amount of ethyl acetate. Yield 84 mg (23 percent d. Th.) 5'-0-acetyl-N-hydroxy-2,2 ^{Zt l} -cyclocytidin hydrobromide.

UV absorption spectrum: A ^H 2 ° (Ex 10 " ⁵ ), 253 mu (16.4).

Max

Nuclear Magnetic Resonance Spectrum (δ): 8.34 ppm, 8.12 ppm (d.d., 1H, Cg-H),

5.46 ppm (d, 1H, C ₂ -H), 2.15 ppm (s, 3H, methyl proton) C ₁₁ H ₁₄ O ₆ N ₅ Br; CH, N

calc .: 36.28, 3.87 11.53 found: 36.41 4.10 11.26.

509833 / 0S4S

Example 11

. . 4th
278 mg (1 mmol) of N -hydroxy-2,2'-cyclocytidine hydrochloride

are dissolved in 25 ml of anhydrous dimethylformamide, mixed with 204 mg (2 mmol) of acetic anhydride and reacted for 18 hours at 80 ⁰ C. When the reaction has ended, the solvent is distilled off under reduced pressure and the residue is washed with diethyl ether. The product, which is insoluble in diethyl ether, is dissolved in a small amount of methanol and placed on a column of dimensions 5 × 60 cm filled with silica gel for chromatographic purposes. The product fractions obtained after eluting with a mixture of ethyl acetate, diethyl ether and water in a volume ratio of 60:30:10 are combined and evaporated. Yield 48 mg (15 percent of theory)

4th
Th.) 5 * -O-Acetyl-N -hydroxy ^^ '- cyclocytidine hydrochloride.

After recrystallization from ethanol, the product shows the following Characteristics:

UV absorption spectrum: ^. ^H 2 ° (£ x10 ~ ³ ), 253 mu (15.8).

Max

H ₁₄ O ₆ N ₃ Cl; 41 C. 4th H 1 N 14th ber .: 41 , 33 4th , 41 1 3, 04. found: , 32 , 60 3, 1

Example 12

This example shows the preparation of a compound of the invention in the form of the hydroiodide from the corresponding hydrochloride explained by ion exchange.

20 ml of an aqueous solution with 1 g of S'-O 2,2'-cyclocyt ± din hydrochloride are added to a 50 ml

S09B33 / 064S

Ion exchanger in the iodide form (Dowex-1) filled column given. The eluate and the subsequent aqueous washing solutions are combined and dried under reduced pressure evaporated. The residue is recrystallized from ethanol. The 5'-0-acetyl-N-hydroxy-2,2'-cyclocytidine hydroiodide is obtained.

Example 13

277 mg (1 mmol) N-amino-2,2'-cyclocytidine hydrochloride are dissolved in 25 ml of anhydrous dimethylacetamide, treated with 204 mg (2 mmol) of acetic anhydride and 20 hours at 60 ⁰ C. ditched. After the reaction has ended, the solvent is distilled off under reduced pressure and the residue is washed with diethyl ether. The product, which is insoluble in diethyl ether, is dissolved in a small amount of a mixture of isopropanol, acetic acid and water in a volume ratio of 5: 1: 2 and chromatographed on a column filled with 70 g of silica gel for chromatographic purposes. The product fractions obtained after eluting with a mixture of isopropanol, acetic acid and water in a volume ratio of 5: 1: 2 are combined and evaporated. The residue is recrystallized from acetone. Yield 131 mg (42 percent of theory) of 5'-0-acetyl-N -amino-2,2 ^I -cyclocytidine hydrochloride. ^H 2 ° (£ χ 1 '(T ³ ), 263 mu, 273 (s) mu, 285 (s) mu "max '." ■

(14.2, 11.4, 5.0).

) ΌΗ ~

max 257 mu, λ ^ _3x 267 mu, 285 (s)

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C ₁₁ I. 41 C. H 74 N 58 41 , 45 4, 72 17, 59 , 66 4, 17, I ₁₅ O ₅ N ₄ C1; ber .: found:

Example 14

278 mg (1 mmol) of N-hydroxy-2,2'-cyclocytidine hydrochloride are dissolved in 20 ml of anhydrous pyridine with stirring, and 3 mmol of acetic anhydride are added dropwise over 30 minutes while stirring. After a while the reaction mixture becomes clear. After I6stündiger reaction, the reaction mixture with 30 ml of ethanol is added and heated for 30 minutes at 50 ⁰ C. The reaction mixture is then evaporated under reduced pressure and the residue is dissolved in anhydrous ethanol. The solution is evaporated to separate the remaining pyridine. This process (addition of "anhydrous ethanol and evaporation) is repeated three times. The semi-solid residue is recrystallized from a little methanol containing ethyl acetate. Yield 202 mg (62 percent d. Th.) 3 ^1, 5'-di-acetyl-0- N -hydroxy-2,2'-cyclocytidine When a mixture of a 5 percent solution of hydrogen chloride in methanol and ethyl acetate is used for recrystallization, the hydrochloride is obtained.

Example 15. ^l

287 mg (1 mmol) of N -hydroxy-2.2 ^l -cyclocytidine hydrochloride are suspended in 20 ml of anhydrous diethylamine while stirring, and 3 mmol of benzoic acid are added within 30 minutes while stirring

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anhydride added. The reaction mixture after some time klar..Nach I6stündiger reaction ', 3 ml of methanol was added, and the mixture is heated for 30 minutes at 50 ⁰ C. The reaction mixture is then evaporated under reduced pressure and the residue is dissolved in anhydrous ethanol. The solution is evaporated to separate off the remaining diethylamine. This procedure (addition of anhydrous ethanol and evaporation) is repeated three times. The semi-solid residue is recrystallized from ethyl acetate containing a little methanol. Yield 234 mg (52 percent of theory) of 3 ', 5'-di-0-benzoyl-N -hydroxy-2, 2' -cyclocytidine. When using a mixture of a 5 percent solution of hydrogen chloride in methanol and ethyl acetate for recrystallization, the corresponding hydrochloride is obtained. The product has the same properties as the product prepared in example 6;

Example 16

288 mg (1 mmol) of N-hydroxy-2, ^ '- cyclocytidine hydrochloride are suspended in 20 ml of anhydrous pyridine and 3 mmol of acetyl chloride are added within 30 minutes while stirring. After a while the reaction mixture becomes clear. After 20 hours of reaction, 3 ml of methanol are added and the reaction mixture is left to stand at room temperature for 2 hours. After working up according to Example 14, the 3 ¹ , 5'-di-0-acetyl-

4th
N -hydroxy ~ 2,2'-cyclocytidine obtained. The hydrochloride is prepared according to Example 14. The product has the same properties as the product prepared in Example 14.

509833 / 06A5

Example 17

277 mg (1 mmol) of N-amino-2,2'-cyclocytidine hydrochloride suspended with stirring in 20 ml of anhydrous pyridine, slowly admixed with 1.02 g (10 mmol) of acetic anhydride and Left to stand at room temperature for 24 hours. After the reaction has ended, the solvent is distilled off under reduced pressure, the residue is mixed with water and evaporated, this process (addition of water and evaporation) is repeated several times repeated. The semi-solid residue is recrystallized from ethyl acetate containing a little methanol. Yield 195 mg (60 percent of theory) 3 ', 5'-di-0-acetyl-N -amino-2,2'-cyclocytidine. When using a mixture of a 5 percent solution of hydrogen chloride in methanol and ethyl acetate for recrystallization the corresponding hydrochloride is obtained. The product has the same properties as that in Example 8 manufactured product.

SG9833 / 0S4S

Claims (24)

Claims \1. N-substituted 2,2'-0yclocytidine compounds of the general Formula I. • N-Y (ι) 1 in which Y is a hydroxyl or amino group and R and R, which are the same or different, hydrogen atoms or Represent acyl radicals of the general formula R-C-, ■ '0 ■ in which R is an unbranched or branched alkyl radical with 1 Ms 19 carbon atoms, one optionally with at least one halogen atom, an alkyl radical with 1 to 4 carbon atoms, an alkoxy radical having 1 to 4 carbon atoms or a nitro group substituted aryl radical with 6 to 10 carbon atoms, one optionally in the aryl part by at least one halogen or . atom / an alkyl radical with 1 to 4 carbon atoms substituted aralkyl radical with 7 to 12 carbon ■ ^ ■ ■ lenstoffatomen, a cycloalkyl radical with 4 to 10 carbon atoms or a polycyclic hydrocarbon radical of the cage type having 7 to 14 carbon atoms, where min- ■ 1 ?
at least one of the radicals R and R is an acyl radical, and their salts with acids. . "· 2. 3 ^f -0-acyl-N -hydroxy-2,2'-cyclocytidine .. '609833 / 0S4S 3 .. 3'-0-Benzoyl-N-hydroxy-2,2'-cyclocytidine. 4. 3'-O-acetyl-N -hydroxy-2,2 ^! -cyclocytidine. 5. 3 »-O-pelmitoyl-N -hydroxy-2,2'-cyclocytidine. 4 6. 3'-O-p -chlorobenzoyl-N -hydroxy-2,2-cyclocytidine. 7. 3 ^s -O-AcyX-N ^ -amimo-2,2 ⁹ -cyclocytidine. 8. 3 ^s O-acetyl-N amino »2 ₈ 2'-cyclocytidine. 9. 5'-O-acyl-N -hydroxy-2,2'-cyclocytidine, 10. 5'-0-Benzoyl-N -hydroxy-2j2 • -cyclocytidine. 11. 5'-O-acetyl-N -hydroxy-2,2'-cyclocytidine. 12. 5'-0-Adamantoyl-N -hydroxy-2,2'-cyclocytidine. ¹ 4
13. 5'-0-p -toluyl- ^N -hydroxy-2,2'-cyclocytidine. 14. 5'-0-Phenylacetyl-N -hydroxy-2,2'-cyclocytidine. 15. 5'-O-acyl-N ^ -amino-2,2 · cyclocytidine. 16. 5'-O-acetyl-N -amino-2,2 · cyclocytidine. 17. 3 ', 5'-Di-0-acyl-N ⁴ -hydroxy-2,2'-cyclocytidines. 5G9Ö33 / 0S4S 18. 3S5'-di-0-acetyl-N -hydroxy ^^ '- cyclocytidine. H
19. 3 ¹ 15'-Di-O-benzoyl-N-hydroxy-2,2'-cyclocytidine. 20. 3 ', 5'-di-0-stearoyl-N -hydroxy-2,2'-cyclocytidine. 21. 3 ¹ ^ '- di-O-caproyl-N -hydroxy-2,2'-cyclocytidine. 22. 3 ^1, 5'-di-O-HCyI N-amino-2,2'-cyclocytidine. 23. 3'15 · Di-0-acetyl-N -amino-2,2'-cyclocytidine. 24. Medicament containing a compound according to claim 1 to 23 and usual carriers and / or diluents and / or auxiliaries. 509833/0545