DD269854A5 - METHOD FOR THE PRODUCTION OF DESAZA-PURINE UNCLEOSIDE DERIVATIVES AND THEIR USE IN NUCLEIC ACID SEQUENCING AND AS ANTIVIRAL AGENTS - Google Patents

Abstract

According to the invention, novel desaza purine nucleosides of the formula (I) are prepared in which, for example: X is nitrogen or a methine group, W is nitrogen or the group R 1, R 2, R 3, R 4, identical or different, hydrogen, halogen, a lower alkyl, hydroxy , Mercapto, lower alkylthio, lower alkyloxy, aralkyl, aralkyloxy, aryloxy or an optionally monosubstituted or disubstituted amino group, R5 is hydrogen or a hydroxy group, R6, R7 are each hydrogen or one of the two radicals R6 and R7 is halogen, a cyano, an acid or an optionally mono- or di-substituted amino group Y hydrogen, a monophosphate, diphosphate or triphosphate group and also possible tautomers and salts and Nucleinsaeuren containing one or more compounds of formula I as a building block. Formula (I)

Description

in the

X, W, R ¹ , R ² , R ³ , R ⁵ , R ⁶ ', R ⁷ ' and R 'have the above angegebono meaning, and optionally eliminates any existing oxygen protecting group and then optionally

a compound thus obtained in which R ⁶ or R ^{7 represents} a hydroxy group, after prior selective protection of the 5'-hydroxy group with a halide, cyanide or azide in a known manner in a compound of formula I in which R ^e or R ^{7 is} halogen , a cyano or an azido group, converted or in a known manner to a compound of formula I, in which

R ⁶ or R ^{7 is} hydrogen, deoxygenated

or a compound of the formula I in which R ⁶ or R ^{7 is} an azido group is converted in a known manner to give a compound of the formula I in which R ⁶ or R 'is an amino group

and, if desired, compounds of the formula I in which Y is hydrogen are then converted into the mono-, di- or tri-phosphates in a known manner and, if desired, free bases or acids obtained are converted into the corresponding salts or salts obtained into the corresponding free bases or acids ,

2. Use of compounds according to claim 1, characterized in that they are used in DNA sequencing.

3. Use of compounds according to claim 1, characterized in that they are used for the preparation of medicaments.

Field of application of the invention

The invention relates to a process for the preparation of desaza purine nucleoside derivatives and to the use of these nucleoside derivatives in the sequencing of nucleic acids and as antiviral agents.

Characteristic of the known state of the art

The building blocks of the nucleic acids contain as glycosidic component either the ß-D-ribofuranosyl or its 2'-deoxy derivative. In addition to these aglycic residues, modified nucleoside antibiotics have found modified D-ribofuranosyl derivatives. So z. Cordycepin, which can be isolated from the culture filtrates of Cordyceps militaris, the monosaccharide Cordycepose. In addition to this 2'- or 3'-deoxy derivative of the ribonucleosides, 2 ', 3'-dideoxynucleosides have been synthetically synthesized some time ago. They have an antiviral action and can specifically inhibit the proliferation of retroviruses by inhibiting the enzyme reverse transcriptase (compare Proc Natl Acad., See, USA 83 [1986] 1911 and Nature 326 (1987) 773). Of particular therapeutic interest is the inhibitory effect on the HIV virus, the causative agent of AIDS. However, they have the disadvantage that they are inhibitors of cellular DNA polymerase, so that they act cytotoxic. Furthermore, they can be deactivated by cellular enzymes.

Object of the invention

The aim of the invention is the provision of novel compounds with valuable pharmacological properties, in particular with antiviral activity, which are free of side effects and in particular have no cytotoxic effect.

Explanation of the essence of the invention

The invention has for its object to find new compounds with the desired properties and processes for their preparation.

According to the invention, novel desaza purine nucleoside derivatives of the general formula I

R ¹ R ³

(I)

made in the

X is nitrogen or a methine group,

W nitrogen or the group "^ C-R ^

R ¹ , R ² , R ³ , R ⁴ , which may be the same or different, are hydrogen, halogen, lower alkyl, hydroxy, mercapto, lower alkyl, lower alkyloxy, aralkyl, aralkyloxy, aryloxy or optionally mono- or di-substituted amino group,

R ^{5 is} hydrogen or a hydroxy group,

R ⁶ , R ^{7 are} each hydrogen or one of the two radicals R ⁶ and R ^{7 is} halogen, p '- is cyano, an azido or an optionally monosubstituted or disubstituted amino group,

wherein one of R ⁶ and R ^{7 may} also benefit from a hydroxy group when X is a methine group, and also R ⁵ and R ⁷ together may represent another bond between C-2 'and C-3' and Y is hydrogen, a monophosphate -, Diphophat- or Triphospr MgrLppe presents, 'and possible tautomers and salts and nucleic acids containing compounds of formula I as a building block.

The lower alkyl radicals in the definition of the substituents R ¹ , R ² , R ³ and R ⁴ can be saturated or unsaturated, straight-chain or branched and contain 1-7, preferably 1-4, carbon atoms. This definition of alkyl radicals also applies to the alkyl radicals which occur in the definitions of the lower alkylthio and lower alkoxy radicals. Very particularly preferred are the methyl and the ethyl group.

Halogen in the definition of the substituents R ', R ² , R ³ , R ⁴ , R ^e and R ⁷ is understood as meaning fluorine, chlorine, bromine and iodine.

In the definitions of the substituents R ^1, R ^2, R ³ and R ⁴ aralkyl or aralkoxy occurring residues containing an alkyl group having 1 to 5, preferably 1-3 carbon atoms, the one or more times with an aromatic radical, for example Phenyl or naphthyl radical. The aromatic radicals may in turn be mono- or polysubstituted by an alkyl or alkoxy group each having 1-3 carbon atoms. Particularly preferred is the benzyl group.

Particularly preferred aryloxy radicals in the definition of the substituents R ¹ , R ² , R ³ and R ⁴ are phenyloxy radicals which may optionally be monosubstituted or polysubstituted by further substituents, for example nitro, alkyl and alkoxy groups.

The amino group occurring in the definition of the substituents R ¹ , R ² , R ³ , R ⁴ , R ^e and R ⁷ , which may optionally be monosubstituted or disubstituted, preferably contains, as possible substituents, alkyl groups having 1-5, preferably 1- 3 carbon atoms, which in turn may be substituted by alkoxy groups, halogen or optionally mono- or poly-substituted amino groups. These substituents may also represent an aralkyl group. The two nitrogen substituents may together also represent an alkylidene, preferably a methylidene radical, which in turn may be substituted by alkoxy, substituted amino groups or halogen. A most preferred substituent of this type is the dimethylaminomethylidene group.

The monophosphate group is the group -PO (OH) ₂ , the diphosphate group, the group -P ₂ O ₃ (OH) ₃ and the triphosphate group means the group-P ₃ O ₆ (OH) ₄ .

Possible salts are, in particular, alkali metal, alkaline earth metal and ammonium salts of the phosphate groups. As alkali salts are

Lithium, sodium and potassium salts are preferred. The alkaline earth salts are in particular magnesium and calcium salts

Question. According to the invention, ammonium salts are understood as meaning salts which contain the ammonium ion, which may be substituted up to four times by alkyl radicals having 1-4 carbon atoms and / or aralkyl radicals, preferably benzyl radicals. The substituents here may be the same or different. The salts of the phosphates can be converted in a known manner into the free acids.

The compounds of the formula I may contain basic groups, in particular amino-groups, which can be converted into acid addition salts with suitable acids. Examples of suitable acids are: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, fumaric acid, succinic acid, tartaric acid, citric acid, maleic acid, maleic acid or methanesulfonic acid.

The compounds of general formula I are novel compounds. They can be prepared in analogy to known, related compounds. To be particularly useful for the preparation of the compounds of formula I has proven a Verfahreii in which a compound of formula II

in the

X, W, R ¹ , R * and R ^{3 have} the abovementioned meaning,

with a compound of formula III

Q 'is -O

(Ill),

in the

R ^{6 has} the meaning given above,

R ⁶ , R ^{7 are} each hydrogen or one of the two radicals R ⁶ and R ⁷ 'is an azido or a hydroxy protected by an oxygen protecting group j,

R 'is an oxygen protecting group and

Z is a reactive group

to compounds of formula IV

(IV)

in the

X, W, R ', R ² , R ³ , R ⁶ , R ⁶ ', R ⁷ 'and R' have the abovementioned meaning, and optionally eliminates any existing oxygen protecting groups and then optionally

a compound of this type in which R ⁶ or R ⁷ denotes a hydroxy group, after prior selective protection of the

B'-hydroxy group with a halide, cyanide or azide in a known manner in a compound of formula I, in cer R ⁶ or R ^{7 is} halogen or a cyano or azido group, converted or in a known manner to a compound of formula I, in R ⁶ or R ^{7 is} hydrogen, deoxygenated

or a compound of the formula I in which R ⁶ or R ^{7 is} an azido group is reduced in a known manner to give a compound of the formula I in which R ⁶ or R ⁷ denotes an amino group

and, if desired, compounds of the formula I in which Y is hydrogen are then converted into the mono-, di- or triphosphates in a known manner

and, if desired, free bases or acids, respectively, which are obtained in the corresponding sows or salts obtained, the corresponding free bases, for example. Acids converted.

The compounds of the formula II are reacted with the compounds of the formula III, particularly advantageously under phase transfer conditions. Under the conditions of phase transfer catalysis, the Gasoen be dor ^p r sleeves Il transferred to a corresponding anion, sodium, for example by 50% aqueous sodium hydroxide solution The resulting Anun is a Phasantransferkatalysator, for example TrU2- (2-m6thoxyethoxy) dthyl | amine hydrophobic, and in transports the organic phase in which it reacts with the reactive compound of formula III.

Suitable reactive groups Z in the compounds of general formula III are preferably halogen radicals and alkoxy groups. The hydroxy groups of the sugar radical were in this reaction in the usual way by those skilled in familiar oxygen protecting groups, for example, toluoyl, benzoyl or acetyl groups, goschützt. After completion of the reaction, the oxygen-protecting groups can be eliminated again in a known manner under alkaline conditions. It is convenient to use a 1M methanolic solution of methanolate. It may be convenient to keep the radicals iuch h \ R ^2, R ³ and R ^{4 is} protected during the reaction by suitable protecting groups.

Another advantageous method for preparing the compounds of formula IV is the solid-liquid phase transfer method using solid, powder-formulated potassium hydroxide, the above-mentioned cryptand, and the compounds of formulas II and III in an aprotic solvent.

Compounds of Forr : r ι1 in which R ⁶ or R ^{7 is} halogen or an azido group, are preferably prepared by starting from compounds of formula I, in which R ^e or R ^{7 represents} a hydroxy group. The hydroxy group in 5'-Ste! Li: ng is initially selectively protected. Again, methods known to those skilled in the art are available. For example, the 4,4'-dimethoxytriphenyl methyl group has been found to be useful in nucleotide chemistry. This reaction can again be easily cleaved off by mild acid hydrolysis after the reaction has taken place, while the acid-labile (ilycosidic bond under this reaction) is not hydrolyzed The reaction of the niocoside to be protected with the oxygen protecting group reagent for dib 5'-hydroxy group is carried out in a suitable organic solvent, pyridino-pyridine, with a slight excess of the oxygen-protecting group reagent and optionally a suitable auxiliary base, for example N-ethyldiisopropylamine The protected compound of formula I is treated with a halide, preferably an alkali halide or an organic halide, ozv / with an azide, suitably with an alkali azide, in a well-known manner, where the OH group with C-3'atom is nucleophilically substituted by the halide or azide H ^e or R ^{7 is} a hydroxy group b According to known methods, even after previous protection of the 5'-hydroxypropionate in the above manner, it is possible to deoxygenate by known methods to give compounds of the formula I in which R ^e and R ⁷ signify hydrogen. Hiorzu is the compound of the general formula I in which R ⁶ or R ^{7 represents} a hydroxy group and in which the 5'-Hydrox>'group has been protected in the above manner and also carry other functional radicals protecting groups, first η a 3' Transferred O-thiocarbonyl derivative, which is then radically reduced with tributyltin hydride. Such methods for deoxygenating 2'-deoxynucleorides to 2 ', 3'-dideoxynucleosides are known to those skilled in the art. The method of Barton deoxvation has proven to be particularly favorable (J. Chem. Soc, Perkin Trans. I [1975] 1574).

Compounds of the formula I in which R ⁶ or R ⁷ denotes an amino group are advantageously prepared by reacting compounds of the formula I, it. this radical R * or R ^{7 represents} an azido group, reduced. This reduction of the azido group to the amino group can according to various, well-known. Methods are done. The reduction with hydrogen has proved to be particularly advantageous for a palladium-carbon catalyst.

The phosphate groups are introduced into compounds of the general formula I in which Y is hydrogen, in known manner. The monophosphates are obtained, for example, by phosphorylating compounds of the formula I in which Y represents hydrogen with phosphoryl chloride in trimethyl phosphate. The triethylammonium salts obtained in this way can be converted in a known manner into other salts by means of conversions. The di- or triphosphates are obtained by known methods, preferably from the monophosphates by reaction with o-phosphates or pyrophosphates. Their various salts can also be prepared by known methods. The compounds of the formula II are known compounds or can be prepared in analogy to known compounds. Such preparation methods are described, for example, in Chemische Berichte 110 (1977) 1462, J. Chem. Soc. 1960,131 and Tetrahedron Letters 21 (1980) 3135.

The compounds of the formula III are also known compounds. Heretofore not envisaged compounds can be prepared in complete analogy to the known compounds. The preparation of such a compound is described for example in Chem. Ber. 93 (1960) 2777 and iiynthecis 1984, 961, respectively.

The novel compounds of the present invention have valuable pharmacological properties. In particular, inhibition of the enzyme reverse transcriptase prevents the proliferation of retrovirun, i. H. the substances according to the invention have, in particular, cytostatic and antiviral properties.

The compounds according to formula I do not have the disadvantages of the known compounds. S 'act antivirally without being cytotoxic.

The substances of the formula I according to the invention can also be used advantageously in DNA sequencing according to Sanger. In particular, the sequencing of d (GC) -rich DNA fragment is complicated by the formation of secondary structures, JIo lead to a band compression in the range of d (GC) clusters. The reason for this lies in the Hoogsteen base pairing of guanosine molecules. Substitution of α'-D -oxyalkyne triphosphate by compounds of the present invention in which R ^{e introduces} a hydroxy group largely eliminates band compression. The compounds of the formula I according to the invention in which R ⁶ and R ^{7 are} hydrogen are used in the Sanger DNA sequencing as chain terminators instead of the known 2 ', 3'-dideoxy compounds. Nucleic acids which comprise one or more compounds of the formula I as building block can be prepared by known processes (for example Nucleic Acids Research Vol. 4, No. 5,1986, p. 2319ff.). They also arise, for example, in DNA sequencing. Are used as building blocks compounds of formula I, in which R ^e a

Hydroxy group, so a Nucloinsaure can have several such building blocks; If the building block used is a compound of the formula I in which R ^{6 is} hydrogen, such a building block can be incorporated only once, namely at the end of the chain. The inventive. Nucleic acids are made up of from 2 to 1000, preferably 8 to 50, nucleotide units. Particular preference is given to nucleic acids having 15-30 nucleotide units.

These nucleic acids can also be used as antiviral agents. As a so-called anti-sense nucleic acid, this nucleic acid hybridizes with the ssDNA / RNA of the virus and complicates the transcription to the viral DNA. Such nucleic acids can be used as a middle! against AIDS because they are not or only with difficulty degraded by cell-specific restriction enzymes.

For the preparation of medicaments, the substances of the general formula I, their pharmacologically acceptable salts or nucleic acids containing them are mixed in a manner known per se with suitable pharmaceutical carriers, flavors, flavors and dyes and shaped, for example, as tablets or dragees or with the addition of appropriate auxiliary stoppers in water or oil, such. As olive oil, suspended or dissolved.

The inhalation substances can be administered in liquid or solid form, either enterally or parenterally. Water is preferably used as the injection medium, which contains the additives customary in injection solutions, such as stabilizers, solubilizers or buffers.

Such additives are z. Tartrate and citrate buffers, ethanol, complexing agents (such as ethylenediaminetetraacetic acid and its non-toxic salts) and high molecular weight polymers (such as liquid polyethylene oxide) for viscosity control. Solid carriers are z. As starch, lactose, mannitol, methylcellulose, talc, fumed silica, high molecular weight fatty acids (such as stearic acid), gelatin, agar-agar, Cilciumphosphat, magnesium stearate, animal and vegetable fats and solid high molecular weight polymers (such as Polyethyienrjlykole). If desired, preparations suitable for oral administration may contain flavorings and sweeteners.

The compounds according to the invention are usually administered in amounts of 1-100 mg, preferably 2-80 mg per day and per kg of body weight. It is preferred to distribute the daily dose to 2-5 applications, with each application 1-2 tablets having an active ingredient content of 5-1 GOOmg been administered. The tablets may also be retarded, reducing ιί: <»number of applications per day to 1-3. The active ingredient content of the delayed-release tablets can be 2O-2OOOmg. The active ingredient may also be given by injection one to eight times a day or by continuous infusion, with amounts of 50-4000 mg / day usually being sufficient.

embodiments

The invention is further illustrated by the following examples.

Example 1 2-Amino-7-desflza-2 ', 3'-dideoxy-9-β-D-ribofuranosyl-purin-6-one

a) 2 - [(4,4'-Dimethoxytriphenylmethyl) amino] -7-deaza-2'-deoxy-5'-O- (4,4'-dimethoxytriphenylmethyl) -9-.beta.-D-ribofuranosylpurin-6-one

1, Og (3.8 mmol) of 7-deaza-2'-deoxyguanosine is evaporated twice with dry pyridine and suspended in 20 ml of pyridine. 4.0 g (11.8 mmol) of 4,4'-dilethoxytriphenylmethyl chloride and 2.5 ml (14.6 mmol) of Hünig's base (N-ethyldiisopropylamine) are added and the mixture is stirred at room temperature for 3 hours.

The reaction mixture is then added to 150 ml of 5% strength aqueous NaHCO 3 solution and extracted two times, each with 150 ml of CH ₂ Cl ₂ . The combined organic extracts are dried over Na ₂ SO ₄ , filtered and chromatographed on silica gel 6OH (column 10 × 4 cm, CH ₂ Cl ₂ / acetone [9: 1]). After concentrating the main zone, the residue is dissolved in a little CH ₂ Cl ₂ and added dropwise to a mixture of n-hexane / ether (1: 1). After filtration, 2.04 g (61%) of the desired colorless, amorphous compound are obtained. - TLC (silica gel, CH ₂ Cl ₂ / acetone [8: 2]): R _F = 0.7. UV (MeOH): A _mix = 2 / 2.283 nm (Sch.) (Ε = 18800, 16400). -

'H-NMR ([D ₆ ] DMSO): δ = 1.75 (m, 2'-H _b ), 1.86 (m, 2'-H ₁ ), 3.09 (m, 5'-H ), 3.79 (m, 4-H), 4.10 (m, 3'-H), 5.19 (d, 3'-OH, J = 4.3 Hz), 5.61 (pt, TH, J = 6.5Hz), 6.16d, 6-H, J = 3.5Hz), 6.62 (d, 5-H, J = 3.5Hz), 10.35 (s, NH). Cs ₃ H ₅₀ N ₄ Oe (871.0) Ber. C 73.09 H 5.79 N 6.43 Gef. C 73.02 H 5.98 N 6.34

b) 2 - ((4,4'-Dimethoxytriphenylmethyl) amino) -7-deaza-2'-deoxy-3'-O-phenoxythiocarbonyl-5'-O- (4,4'-dimethoxytriphenylmethyl) -9-β- D-ribofuranosyl-purine-6-one

A suspension of 1.0 g (1.1 mmol) of the compound from 1 a) in 15 ml of dry acetonitrile is mixed with 300 mg (2.5 mmol) of p-dimethylaminopyridine and 300 ml (2.2 mmol) of phenoxythiocarbonyl chloride and stirred at room temperature for 16 hours. The reaction mixture is concentrated by evaporation and the residue is chromatographed on silica gel 6OH (column 10 × 4 cm, CH ₂ Cl ₂ / acetone [8: 2]). The residue obtained by evaporating the H 2 O is dissolved in a little CH ₂ Cl ₂ and precipitated by dropwise addition into a mixture of n-hexane / ether (1: 1), giving 0.99 g (89%) of colorless amorphous substance. - TLC (silica gel, CH ₂ Cl ₂ / acetone [8: 2]): R _f = 0.8. -UV (MeOH): A _mlx = 269.282nm (Sch.) (Ε = 19300, 16000) .- ¹ H-NMR (ID ₆ IDMSO): δ = 2.06 (m, 2'-H _b ), 2 , 34 (m, 2'-H _a ), 3.26 (m, 5'-H), 4.25 (m, 4'-H), 5.61 (m, 3'-H and 1'-H) , t.3 (d, 6-H, J = 3.5Hz), 6.67 (d, 5-H, J = 3.5!! *), 10.41 (s, NH).

C ₆₀ H ₅₄ N ₄ O ₉ S (1007.2) Btr. C71.77 H 5.40 N 5.56 S3.18

C 71.26 H 5.43 N 5.52 S 3.11

c) 2 - ((4,4'-Dimethoxytriphunylmethyl) amino) -7-diaza-2'-3'-dideoxy-5'-O- (4,4'-dimethoxytriphenylmethyl) -9-pD-ribfuranosyl-purine 6-one

ROOmg (0.5 mmol) of the compound from 1 b) in 2OmI freshly distilled toluene are mixed with 30mg (0.2 mmol) of 2,2'-azo-bls- (2-meihylpropionsäurenitril) and 300μΙ '1.1 mmol) tributyltin hydride and Stirred under argon at 80 ° C. for 3 hours (TLC check, CHCl ₃ / methanol [97: 3]). After complete reaction, the reaction mixtures are concentrated by evaporation and the residue is suspended on silica gel 60 H (column 30 × 4 ci), ChClj / methanol After evaporation of the main zone and absorption in a little CH ₂ Cl ₂ , 320 mg (75%) of the desired amorphous, colorless compound are precipitated by dropwise addition into n-hexane / ether! - TLC (silica gel, CH ₂ ) ₂ CI ₂ / methanol [95: 5]): R _F = 0.5 ¹ H-NMR ([D ₆ ] DMSO): δ = 1.63, 1.00 (2 m, 2'-H and 3 ') -H), 3, O ^- * (m, 5'-H), 4.06 (m, 4'-H, 5.43 (m, 1'-H), 6.11 (d, 6-H , J = 3.5Hz), 6.65 (d, 5-H), J = 3.5Hz), 10.34 (s, NH).

d) 2-Amino-7-deaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosylpurine-6-one

300 mg (0.35 mmol) of the compound from 1 c) are dissolved in 10 ml of 80% strength acetic acid »and stirred for 15 minutes at room temperature. The solvent is then removed in an oil pump vacuum and the residue is evaporated several times with water. The crude product is chromatographed on silica gel 6OH (column ΊΟ χ 4 cm, CH ₂ Cl ₂ / methanol [9: 1]). The frothy substance obtained by evaporation of the main fraction is crystallized from a little methanol, giving 50 mg (57%) of bare needles of mp 228 ° C. (dec.). -DC (silica gel, CH ₂ Cl ₂ / methanol [9: 1]): R _F = 0.3. UV (MeOH): Jw = 261.281 nm (Sch.) (Ε = 13300.7800) .-

¹ H-NMR ([D ₈ ] UMSO): δ = 1.96 (m, 3'-H ;, 2.08.2.27 (2m, 2'-H, and 2'-H _b ), 3 , 48 (m, 5'-H), 3.97 (m, 4'-H), 4.86 (t, 5'-OH ₁ J = 5.4Hz), 6.12 (pt, 1'- H, J = 5.5Hz), 6.24 (m, NH ₂ and 6-H), 6.92 (d, 5-H, J = 3.5Hz), 10.34 (s, NH). CuH ₁₄ N ₄ O ₃ (250.3) calc. C 52.79 H5.64 N 22.39 Gef. C 52.98 H 5.80 N 22.55

In an analogous manner is obtained via the corresponding 2'-deoxynucleosides and subsequent deoxygenation, as described under c):

A) 3,7-Dldesaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purine UV (0.1 n-HCl): K _mn = 224.274 nm

C ₁₂ H ₁₄ N ₂ O ₂ (218.2) Ber. C 66.0 H 6.4 N 12.8 Gef. C 66.1 H 6.4 N 12.6

B) 3,7-Dldesaza-2 ', 3'-dldesoxy-9-.beta.-D-ribofuranosyl-purine-6-one

UV (methanol): X _m . _x = 264nm (ε = 11600), 282nm (ε = 8000), 295nm (ε = 5200) C ₁₂ H ₁₄ N ₂ O ₃ (234.2) m. C61.5 H6,0 N 11,95 Gef. C 61,3 H 6,1 N 11,8

C) 2-Chloro-6-methoxy-3,7-dideaza-2 ', 3'-dideoxy-9-β-D-ribofuranosyl-purine UV (methanol) A _m "= 271.280 nm

C ₁₃ H ₁₆ N ₂ O ₃ Cl (282.6) Calcd. C 55.2 H 5.3 N 9.9 Gef. C 55.1 H 5.3 N 9.9

D) 6-Amino-3,7-dideaza-2 ', 3'-dideioxy-9-.beta.-D-ribofuranosyl-purine C ₁₂ H ₁₅ N ₃ O ₂ (233.2) Ber. C 63.65 H 6,16 N 17,13

Found C 63.62 H 6.11 N 17.01 UV (methanol) X _mn 271 nm (ε = 12800)

E) 3,7-Dideaza-2 ', 3'-dideoxy-9-β-uro: hofuranosyl-purine-2,6-dione C ₁₂ H ₁₄ N ₂ O ₄ (250.2) Ber. C57. & 5 H5.6 N 11.2

Gef. C 57.50 H 5.7 N 11.2

Example 2

2 - {[(dimethylamino) methylene] amino} -7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-purine-6-one

a) 2 {[(dimethylamino) methylene] amino} -7-desaza-2'-deoxy-9-.beta.-D-ribofuranosyl-purin-6-one 270mg (1.01 mmol) 7-deaza-2'-deoxy- guanosine in 5 ml of dry, amine-free dimethylformamide are mixed with 2 ml (11.7 mmol) of Ν, Ν-dimethylformamide diethyl acetal and stirred for 1 hour at 50 ^° C. under argon. Subsequently, the reaction mixture is evaporated in vacuo and r η 6OH silica gel (column 10 x 4 cm, CH ₂ Cl ₂ / methanol [9: 1 j) chromatographed. Evaporation of the solvent gives the main zone 230 mg (71%) of pale yellow, amorphous substance.

TLC (silica gel, CH ₂ Cl ₂ / methanol [9: 1]): R _F = 0.3. -UV (MeOH): A _m "= 240.311 nm (ε = 18300, 1700) .- ¹ H-NMR ([D ₆ ] IDMSO: δ = 2.15 (m, 2'-Hb), 2.41 ( m, 2'-H,), 3.02.3.15 (s, 2CH ₃ ), 3.52 (m, 5'-H), 3.79 (m, 4'-H), 4.32 in, 3'-H), 4.91 (t, 5'-0H, J = 5.4 Hz), 5.27 (d, 3'-OH), J = 3.5 Hz), 6.34 (d, 6-H, J = 3.5 Hz), 6.45 (pt, 1'-H, J = 7.0 Hz), 7.07 (d, 5-H, J = 3.5 Hz ), 8.56 (s, NH = C), 11.04 (s, NH).

C ₁₄ H ₁₉ N ₆ O ₄ (321.3) Ber. C 52.33 H 5.96 N 21.79 Gef. C 52.48 H 6.14 N 21.69

b) 2 - {[(Dimethylamino) -methylonlamino} -7-de3aza-2'-deoxy-5'-O- (4,4'-dimethoxytriphenylmethyl) -9-.beta.-D-ribofuranosyl-purin-6-one

100 mg (0.31 mmol) of the compound from 2a) are dissolved in 2 ml of dry pyridine, mixed with 170 mg (0.5 mmol) of 4,4'-dimethoxytriphenylmethyl chloride and 0.2 ml (1.2 mmol) of Hünig base and left to stand for 3 hours stirred at room temperature. The reaction mixture is then evaporated and the residue is chromatographed on silica gel 6OH (column 10 × 2.5 cm, solvent CHCl ₃ / methanol [99: 1]). The residue obtained by evaporation of the main fraction is dissolved in CH ₂ Cl ₂ and by dropwise addition to a mixture of n-hexane / ether (1: 1) 160 mkg (84%) of colorless, amorphous substance are precipitated. - TLC (silica gel, CH CI ₂ / methanol [9: 1)): R _F = 0.6. - UV (MeOH): X _011x = 236.311 nm (ε = 38200.18100) .-

¹ H-NMR (IDe! DMSO): δ = 2.23 (m, 2'-H "), 2.42, (m, 2'-H ₁ ), 3.03 (s, CH ₃ ), 3 , 14 Im, 5'-H and CH ₃ ), 3.90 (m, 4'-H), 4.33 (m, 3'-H), 5.34 (d, 3'-OH, J = 4.3Hz), 6.34 (d, 6-H, J = 3.5Hz), 6.49 (pr, 1'-H, J = 6.8Hz), 6.90 (d, 5-H, J = 3.5Hz), 8.58 (s, NH = C), 11.07 (s, NH). C ₃₅ H ₃₇ N ₅ Oe (623.7) Ber. C 67.40 H 5.98 N 11.23 Gef. C 67.31 H 6.00 N 11.17

c) 2 - {((Dimathylamino) methylene] amino} -7-deaza-2'-deoxy-3'-O-phenoxythiocarbonyl-5'-O- (4,4-dimethoxytriphenylmethyl) -9-.beta.-D-ribofuranosyl purine-6-one

900 mg (1.4 mmol) of the compound from 2 b), dissolved in 15 ml of dry CH ₂ Cl ₂ , are mixed with 340 mg (2.8 mmol) of p-dimethylaminopyridine and 250 μl (1.8 mmol) of phenoxythiocarbonyl chloride and stirred at room temperature for 16 hours. The solution is evaporated in vacuo and the residue is chromatographed on silica gel 6OH (column 20 × 4 cm, CHCIs / acetone [7: 3], CHCl ₃ / acetone [6: 4]). The residue obtained by evaporation of the main zone is taken up in a little CH ₂ Cl ₂ and 980 mg (90%) of the desired colorless, amorphous compound are precipitated by dropwise addition into n-hexane / ether (1: 1). -

TLC (silica gel, CHjClj / methanol [95: 5]): R _F = 0.5.UV (MeOH): \ _mix = 235.312nm (ε = 41300, 1400, 12600, 17000) .- ¹ H-NMR ([ D ₆ ] DMSO): δ = 2.73 (m, 2'-H "), 2.97 (m, 2'-H ₁ ), 3.01.3, 10 (s, 2CH ₃ ), 3, 37 (m, 5'-H), 4.33 (m, 4'-H), 5.90 (m, 3'-H), 6.40 (d, 6-H, J = 3.5Hz) , 6.55 (pt, VH), 6.98 (d, 5-H, J = 3.5Hz), 8.58 (s, CH = N), 11.30 (s, NH). O ₁₂ H ₄ IN ₆ O ₇ S (759.9) Ber. C 66.39 H 5.44 N 9.22 S 4.22 Gef. C 66.49 H 5.55 N 9.25 S 4.29

d) 2 - {[(dimethylamino) methylene] amino} -7-desaza-2 ', 3'-dideoxy-5'-O- (4,4'-dimethoxytriphenylmethyl) -9-β-D-ribofuranosylpurine-6 on

500 mg (0.7 mmol) of the compound from 2c), dissolved in 20 ml of freshly distilled toluene, mixed with 25 mg (0.15 mmol) of 2,2'-azobis (2-methylpropionitrile) and 50 μl (1.9 mmol) of tributyltin hydride, and stirred at 8O ⁰ C under argon for 16 hours. The reaction mixture is then evaporated in an oil pump vacuum and the residue is suspended on silica gel 6OH (column 20 χ 4 cm, CH ₂ Cl ₂ / acetone (9: 1), CHCl ₃ MCeUJn [7: 3], CHCl ₃ / acetone [6: 4]). The residue obtained by concentrating the main fraction is dissolved in a little dichloromethane and precipitated by dropwise addition into n-hexane / ether, giving 320 mg (80%) of the desired colorless, amorphous compound

TLC (silica gel, CH ₂ Cl ₂ / methanol [95: 5]): R _F = 0.3. -

UV (MeOH): A _n ,,., = 236.277 (Sch.), 284.312 nm (ε = 37200, 12000, 13 500, 18 000) .-

¹ H-NMR ([D ₆ ] DMSO): δ = 2.02 (m, 3'-H), 2.20, 2.33 (m, 2'-H and 2'-H _b ), 3 , 02,3,13 (s, CH _3), 3.08 (m, 5'-H), 4.17 (m, 4'-H), 6; 31 (d, 6-H, J = 3 , 5Hz), 6.38 (m, 1'-H), 6.92 (d, 5-H, J = 3.5Hz), 8.61 (s, CH = N), 11.07 (s, NH). C ₃₆ H ₃₇ N ₅ O ₇ (607.7) Ber. C 69.18 H 6.14 N 11.52 Gef. C 69.23 H 6.24 N 11.61

e) 2 - {[(Dimethylamino) methylene] amino} -7-desaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purin-6-one

130 mg (0.21 mmol) of the compound from 2 d) are dissolved in 5 ml of 80% acetic acid and stirred for 15 minutes at room temperature. Then the acetic acid is evaporated off in an oil pump vacuum and the residue is chromatographed on silica gel 6OH (column 20 × 2 cm), CH ₂ Cl ₂ / methanol [95: 5]). The residue obtained by concentrating the main fraction is foamed by repeated evaporation with acetone, or 43 mg (67%) of the desired colorless, amorphous compound are obtained.

TLC (silica gel, CH ₂ Cl ₂ / methanol [9: 1]): R _F = 0.5% UV (MeOH): A _mix = 239.282 (Sch.), 311 nm (ε = 17400.10500, 690000) .-

¹ H-NMR ([D ₆ ] DMSO): δ = 2.06 2.32 (m, 2'-H and 3'-H), 3.01.3, 14 (s, 2 CH ₃ ), 3 , 51 (m, 5'-H), 4.00 (m, 4'-H), 4.87 (t, 5'-OH), 6.33 (m, VH and 6-H, J = 3 , 3Hz), 7.05 (d, 5-H, J = 3.3Hz), 8.59 (s, CH = N), 11.02 (s, NH). Ci ₄ H ₁₉ N ₅ O ₃ (305.3) Ber. C 55.07 H 6.27 N 22.94 Gef. C 55.23 H 6.41 N 22.75

Example 3

2-Amlno-6-methoxy-7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-purine

a) 2-Amino-6-methoxy-7-deaza-2'-deoxy-9-.beta.-D-ribofuranosyl-purine

543 mg (10 mmol) of finely powdered potassium hydroxide and 68 mg (0.2 mmol) of tetrabutylammonium hydrogen sulfate in 30 ml of absolute dichloromethane are stirred at room temperature for 15 minutes under nitrogen atmosphere. Then 330 mg (2 mmol) of 2-amino-6-methoxy-7-desa7a-purine (2-amino-4-methoxy-7H-pyrrolo [2,3-d] pyrimidine) are added and the mixture is stirred for a further 30 minutes. After addition of 884 mg (2.2 mmol) of 2-deoxy-3,5-di-O-p-toluoyl-.beta.-D-erythro-pentofuranosyl chloride is allowed to stir for a further 3 minutes. It sucks in insoluble constituents, washed with a little dichloromethane and concentrated μ _. the filtrate to about 10 ml. After addition of 3 ml of 1 M sodium methoxide in methanol, men are allowed to stand for 3 hours

Stir room temperature. After neutralization with acetic acid, the solvent is stripped off, the residue is taken up in water, filtered, and the filtrate is chromatographed from a Dowex (1 × 2OH form, 30 × 3 cm) ion exchange acid (water / methanol 9: 1). After stripping, 260 mg (63%) of colorless crystals are obtained from the main zones.

M.p. 152-154 ⁰ C.

TLC (silica gel, CH _a CI ₂ / MeOH 9: 1) R _f = 0.7.

UV (methanol) K _n = 225.259.25 (ε = 24900.3600.7600).

¹ H-NMR (Id ₉ ! DMSO) O = 6.27 (1H, d, J = 3.7Hz), 6.42, C-H, dd J _r , _r . = 8.4Hz, J _n2b = *, 9Hz), 7.10 (1H, d, J = 3.7Hz) ppm.

' ³ C-NMR (ID (IDMSO) δ = 52.49 (OCH ₃ ), 02.37 (CV), 98.85 (C-5), 119.45 (C-6) ppm.

b) The compound obtained according to a) 2-amino-6-methoxy-7-deaza-2'-deoxy-9-.beta.-D-ribofuranosyl-purine is, as described in Example 1 c), to 2-amino-6 -methoxy-7-desaza-2 ', 3'-dideoxy-9-β D-rii ofuranosyl-purine.

Example 4 2-Amino-6-chloro-7-deaza-2 ', 3'-dideoxy-9-β-D-ribofuranusylpurine

a) The compound is nanoh acetylation of 2-amino-7-desaza-2 ', 3'-dideoxy-9ß-D-ribofluoro osyl-purir.-6-one (prepared according to Example 1 d) according to the in Liebigs Ann. Them. 1987, 15-19 described method produced by halogenation.

b) The resulting crude mixture is allowed to stand for 3 hours with methanolic ammonia solution at room temperature to remove the Acetylschutzgruppe, concentrated to dryness and then chromatographed on silica gel with the eluent chloroform / methanol. After combining the main fractions and evaporation, crystallized from H ₂ O. UV (methanol): X _n ,,, = 235,258,316 (ε = 27,800.4300, 5800).

CnH ₁₃ N ₄ O ₂ Cl (268.7): Ber. C 49.1 H 4.8 N 20.8 Cl 130 Gef. C 49.3 H 4.85 N 20.7 Cl 13.1

Example S

2-Amino-7-decaza-2 ', 3'-didasoxy-9-ß-D-ribofuranosyl-purln

286 mg (1 mmol) of 2-amino-6-chloro-7-des3za-2 ', 3'-dideoxy-9-β-D-ribofuranosyl-purine are dissolved in 25 ml of 70% aqueous methanol, to a suspension of 30 mg pre-hydrogenated Pd / C (10%) in 25 ml of 70% aqueous methanol and hydrogenated until the end of H ₂ uptake. The solvent is removed and crystallized from methanol. Yield 180mg (77%).

C _n H ₁₄ N ₄ O ₂ (234.3) Ber. C 56.4 H 6.0 N 23.9 Gef. C 56.3 H 6.0 N 23.7

UV (methanol): A _n ,,,, = 234.256.314 nm (ε = 30600, 400, 500).

Example 6

2-amino-6-mercapto-7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-purine

536 g (2 mmol) of 2-amino-6-chloro-7-deaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purine and 1.5 g (20 mol) of thiourea are suspended in 30 ml of ethanol and stirred for about 1? Heated to reflux for hours. Then the solvent is distilled off, the residue is taken up in about 25 ml of methanol and chromatographed on silica gel 60 H (Gaule 20 × 3 cm, CH ₂ Cl ₂ / MeOH 9: 1). Evaporation of the main fraction and crystallization from methanol / H ₂ O gives 230 mg (43%) of the thio compound C ₁₁ H ₁₄ N ₄ O ₂ S (266.3) Ber. C 49.6 H 5.3 N 21.0 Gef. C 49.4 H 5.4 N 21.1

UV (methanol): A _m "= 235.271.345 nm (ε = 17600, 1700, 180000).

¹ H-NMR (ID ₆ ! DMSO): δ = 1.9 (m, 3'-H), 2.1 (m, 2'-H _b ), 2.34 (m, 2'-H ₁ ) , 3.50 (m, 5'-H), 3.97 (m, 4'-H), 4.86 (t, 5'-OH), 6.12 (m, -1'-H), 6.24 (m, NH ₂ and 8-H), 6.92 (d, 7-H), 11.1 (s, NH)

Example 7

2,6-diamino-7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-pur.n

268 mg (1 mmol) of 2-amino-6-chloro-7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-purine are treated with ^ UmIg aqueous concentrated ammonia solution and 60 hours at 65 ⁰ C. warmed well closed in a water bath. After evaporating off the solvent, it is chromatographed on a silica gel column, first with CH ₂ Cl ₂ / MeOH (9: 1) (starting material), then with CHCl ₃ / methanol (4: 1). After crystallization from Wassre, 120 mg (48%) of the diamino compound are obtained. C ₁₁ H ₁₆ N ₅ O ₂ (249.3) Ber. C 53.0 H 6.0 N 28.1 Gef. C 53.15 H 5.9 N 28.2

UV (methanol): X _max = 264.284 nm (ε = 9800.8000).

'HNMR ([D ₆ IDMSO): δ = 1.9 (m, 3'-H), 2, 1, 2, 4 (2m, 2'-H ₈ , _b ), 3.4 (m, 5 '-H), 3.3 (m, 4'-H), 4.8 (t, 5'-OH), 5.6 (s, NH ₂ ), 6.2 (dd, 1'-H) , 6.3 (d, 7-H), 6.7 (s, NH ₂ ), 6.9 (d, 8-H).

Example 8

Z-methylthio-e-methoxy ^ -desaza ^ '. S'-dideoxy-S-beta-D-ribofuranosyl-purine

a) 2-methylthio-6-methoxy-7-deaza-2'-deoxy-9-.beta.-D-ribofuranosyl-purine

500mg (2.56mmol) of 4-methoxy-2-methylthio-7H-pyrrolo [2,3-d] pyrimidine and 400mg (1.75mmol) of benzyltriethylammonium chloride dissolved in 20mL of dichloromethane with 20ml of 50% sodium hydroxide solution as the counterphase Vibromischer briefly mixed. 1.2 g (3.1 mol) of 2-deoxy-3,5-di-O- (p-toluoyl) -β-D-erythropentofuranosyl chloride are added in a small amount of dichloromethane and the vibromic acid is continued for 30 minutes. The organic phase will? ".. and the aqueous shaken with dichloromethane. The combined organic extracts are dried with water, washed and dried with sodium sulfate. After filtration, it is evaporated and the residue is dissolved in 100 ml of 1M sodium methoxide in methanol. The mixture is stirred for about 12 hours at room temperature, evaporated, taken up in water and adsorbed on a Dowex 1 -X2 ion exchange column (30 χ 3 cm, OH "form). Elution with water-methanol (1: 1)

leads to a main zone. After evaporation of the solvent is recrystallized from water; Yield 321 mg (40%) fsifalose needles with mp 118 ⁰ C. - DC iKleselgel / CHjC ^ / acetone (8: 2).) R _F = 0.26. UV (methanol): X _m "- 283, 236 nm (e = 13000, 1550) ₁ ¹ H-NMR (IDa) DMSO) IO = 2.20 (m, 2'-H), 2.40 (m, 2'-H), 2.56 (8, CH ₃ S), 3.50 (m, 5'-H ₂ ) 3.81 (m, 4'-H), 4.01 (s, CH ₃ O ), 4.35 (m, 3'-H), 4.90 (1.5-OH, J = 5Hz), 5.29 (d, 3'-OH, J = 4Hz), 6.48 (i.e. , 5 H, J = 4Hz), 6.55 (M'-H ₁ J = 5Hz), 7.47 (d.6-H, J = 4Hz).

Ci ₃ H ₁₇ N ₃ O ₄ S (311.4) Calcd. C50, i5 H 5,50 N 13,50 S 10,30 Gef. C 50,28 H 5,47 N 13,56 S 10,31

b) 2-Methylthio-6-methoxy-7-deaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purine

Is prepared by Deoxigeniorung the obtained according to a) 2'Desoxy compound as described in Example I c). UV (methanol): K _n ^, = 283.236nm (e = 1300, 155500) C ₁ JlIuN ₃ O ₃ S (295.4) Ber. C 52.8 H 5.75 N 14.2 Gef. C 52 JH 5.70 N 14.2

Example 9 6-Methoxy-7-desaza-, 3'-dldesjxy-9-β-D-ribofuronosyl-purine

a) 6-Methoxy-7-deaza-2'-d9soxy-9-.beta.-D-ribofuranosyl-purine

The synthesis of the compound takes place as in Liebigs Ann. Chem. 1985, 1360-1366.

b) The dideoxy derivative can be obtained by deoxygenation of the compound of 9a) as described in Example 1 c). An alternative route is the desulfurization of 2-methylthio-6-methoxy-7-deaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purine from Example 8 also according to Liobigs Ann. Chem. 1985, 1360-1366.

TLC (CHjCl ₂ / MeOH 9: 1): R ₍ = 0.8 UV (MeOH): X _mlx = 261 nm (log [e] = 3.86).

¹ H-NMR (DMSO-d _e ): δ = 2.04 (m, 3'-H); 2.24 (m, 2'- _Hb ); 2.40 (m, 2'-H ₁ ); 3.55 (m, 5'-H); 4.04 (s, OCH ₃ ); 4.07 (m, 4'-H); 4.93 (t, J = ", 5Hz, 5'-OH); 6.47 (dd, J = 4.4 and 6.8Hz, VH); 6.55 (d, J = 3.7Hz, 5-H). : 7.66 (i.e., 1 = 3.7 hr RH); 8.42 (s, 2-H). C ₁₂ H ₁₆ N ₃ O ₃ (249.3) calc. C 57.8 H 6.0 N 16. 8 Gef. C 57.8 Ho ₁ O ¹ IN 16.65

Another way of preparing this compound is described in Example 24 i). Example 10

7-desoza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purin-6-one. The preparation of the compound is via the 2'-deoxy compound as in Liebigs Ann. Chem. 1985, 3120-320 and subsequent deoxygenation as in Example 1c). UV (methanol): A _m , _x = 258.280nm (shoulder), (ε - 9200.6400) TLC (CH ₂ Cl ₂ / MeOH 9: 1): R, = 0.5

¹ H-NMR (DMSO-d ₆ ): δ = 2.00 (m, 3'-H); 2.16 (m, 2'- _Hb ); 2.37 (m, 2'-H ₁ ); 3.49 (dd, J = 4.9 and 11.6Hz, 5'-H); 3.58 (dd, J 4, and 11.6Hz, 5'-H); 4.05 (m, 4'-H); 6.33 (dd, J = 4.2 and 6.9Hz, 1'-H); 6.50 (d, J = 3.5Hz, 5-H); 7.36 (d, J = 3.5Hz, 6-H); 7.90 (s, 2-H). C ₁₁ H ₁₃ N ₃ O ₃ (235.2) Ber. C 56.1 H 5.5 N 17.8 Gef. C 56.0 H 5.3 N 18.0

Another possibility for the preparation of this compound is described in Example 24 j). Example 11

7-deaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purine-2,6-diisocyanate synthesis via the 2-deoxy compound as in Liebigs Ann. Chem. 1985, 3120-320 and subsequent deoxygenation as in Example 1c).

UV (phosphate buffer, pH = 7.0): A ", _ax = 251.280 nm (ε = 10500.7400) C ₁₁ H ₁₃ N ₃ O ₄ 1251.4) C 52.5 H 5.2 N 16.7 Gef. C 52.3 H 5.1 N 16.5

Example 12

2,6-dimethoxy-7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-purine

The derivative was synthesized by phase transfer glycosylation of the corresponding purine followed by deoxygenation as described in Example 1c). UV (methanol): A _max = 257.271 nm (ε = 7 300.7 400) C ₁₃ H ₁₇ N ₃ O ₄ (273.3) Ber. C 55.85 H 6.1 N 15.0 Gef. C 55.7 H 6.1 N 15.1

Example 13

6-amino-7-deaza-2 ', 3'-dideoxy-9-ß-D-ribofuranosyl-purine-2-one

The compound was obtained according to J. Chem. Soc., Perkin Trans. Il 1986,525-530, by phase transfer glycosylation of 2-methoxy-6-amino-7-deaza-purine, subsequent demethylation and final deoxygenation analogous to Example 1 c) , UV (methanol): A _max = 255.305 nm (ε = 7600.7200) C ₁₁ H ₁₄ N ₄ O ₃ (250.2) Ber. C 52.7 H 5.6 N 22.4 Gef. C 52.75 H 5.5 N 22.3

Example 14

The connection was made via the in Llebljs Ann. Chem. 1984, 700-721 2'-deoxy-nucleoside with r.chfolgomlor deoxigenization as described in Example 1 c) synthesized

UV (methanol):. ,,, _n X "224,264,285 nm (shoulder) (c = 22600,10500,6500) CuH ₁ I ₁ N ₄ O] (264 ,?) boron 54.5 H 6.05 N 21, 2 Göf. 64.3 H 6.1 N 21.1

Example 15

2-Amlno-7> dessza-2'-3'-dideoxy-3'-eildo-9 * ß * O * rlbofuranosyl purin-6 * on

The compound was prepared by glycosylation of 2-amino-7-desaza-pur! N-6-one with azido-sugar prepared according to Byatkira / Azhayov (Synthesis 1984,901-963).

UV (methanol): X _m , "= 261.281 nm (shoulder) (ε = 13300.7800) C ₁₁ H ₁₃ N ₁ O ₃ (291.3) Ber. C 45.3 H 4.45 N 33.65 Gef. C 45.4 H 4.3 N 33.4

Example 16

3,7-Didesaza-2 ', 3'-dldesoxy-3'-azido-9-ß-D-ribofuranosyl-purln

The compound was prepared by ribosidation of 3,7-didesaza-purine with domido-7-sugar prepared according to Byatkina / Azhayov (Synthesis 1984, 961-963)

UV (methanol): A _n ,,, = 224.274nm CuH ₁₃ N ₆ O ₂ (259.2) Ber. C 55.55 H 5.0 N 27.0 vessel C 55.4 H 5.1 N 26.8 Example 17

6-amino-8-aza-7-deaza-2 ', 3'-dldesoxy-9-.beta.-D-r-bofuranosylpurine (4-amino-1- (2-deoxy-.beta.-D-erythro-pentofuranosyl) -1H- pyrazolo (3 ₍ 4-d) pyrimidine)

a) 4-Benzoylcimino-1 - (2'-deoxy-9-.beta.-D-3-trifluoropentofuranosyl-5'-O- (4,4'-dimethoxytriphenylmothyl) -1H-pyrazolo | 3,4-dlpyrimidine

6-Amino-8-aza-7-deaza-2'-dnsoxy-β-D-ribofuranosyl-purine was prepared as described in HoIv. Chim. Acta 68, 563-570 (1985). The benzoylation of the 4-amino group and the subsequent introduction of the Dimothoxytritylschutzgruppo was carried out analogously to known methods.

b) 4-Benzoylamino-1- (2'-deoxy-β-D-erythro-pentofuranosyl) -5'-O- (4,4'-dimethoxytriphenylmethyl) -3'-O-phenoxythiocarhonyl-1H-pyrazolo (3, 4-d] pyrimidine

200 mg (0.3 mmol) of the product from Example 17 a) were in 4 ml of acetonitrile with 82 .mu.Ι (0.6 mmol) Phenylchlorothiocarbonat at room temperature for 16 hours in the presence of 90mg (0.75 mmol) of 4- (dimethylamino) pyridine umgesotzt , After chromatographic purification (silica gel, dichloromethane / ethyl acetate (95: 5), 150 mg (63%) of the product were isolated TLC (silica gel, CH ₂ Cl ₂ / ethyl acetate, 95: 5): R, = 0.4

¹ H-NMR (ICIDMSO): δ = 3.26 (m, 5'-H), 3.69 (s, 2 x OCH ₃ ), 4.45 (m, 4'-H), 5.98 ( m, 3'-H), (s, 3-H), 8.78 (s, 6-H), 11.72 (s, NH).

c) 4-Benzoylamino-1- (2 ', 3'-dideoxy-9-.beta.-D-glycero-pentofuranosyl) -5'-O- (4,4'-dimethoxytriphonylmethyl) -1H-pyrazolo [3,4 · DJ-l-pyrimidines

200mg (0.25mmol) of the product of Example 17b) were after Barton in 7ml toluene with 150μΙ (0.55mmol) tri-N-butylstannane and 15 mg of 2,2'-azobis (2-methylpropionitrile) for one hour at 80 ⁰ C deoxygenated under argon. Chromatography (silica gel, dichloromethane / ethyl acetate 95: 5) gave 120 mg (75%) of the product (colorless, amorphous). TLC (silica gel, CH 2 Cl 2 / ethyl acetate, 95: 5): R, * - 0.3.

₁ H-NMR (ID ₆ ) DMSO): δ = 2.16 (m, 3'-H), 2.49 (m, 2'-H), 2.99 (m, 5'-H), 3 , 65.3,6C '', 2 x OCH ₃ ), 4.32 (m, 4'-H), 6.39 (m, VH), 8.41 (s, 3-H), 8, 80 (s, 6-H), 11.66 (s, NH).

d) 6-Amino-8-aza-7-deseza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranosyl-purine (4-amino-1- (2', 3'-ciidesoxy-.beta.-D glycero-pentofuranosyl) -1H-pyrazolo (3,4-d] pyrimidine)

a) 300 mg (0.47 mmol) of the product from Example 17c) were dissolved in 40ml saturated with ammonia methanol at 6O ⁰ C for 4 hours, and evaporated to dryness. 200 mg (81%) of 4-amino-1- (2 ', 3'-dideoxy-β-D-glycero-pentofuranosyl) -5'-O- (4,4'-dimethoxytriphenylmethyl) -1 H -pyrazolo [ 3,4-d) pyrimidine as a colorless foam after chromatography on silica gel (dichloromethane / acetone 7: 3).

TLC (silica gel, CH 2 Cl 2 / acetone, 8: 2): R ₁ = 0.25.

¹ H-NMR (ID ₆ ) DMSO) = O = 2.16 (m, 3'-H), 2.45 (m, 2'-H), 2.99 (m, 5'-H), 3 , 69,3,70 (2s, 2 x _OCH3), 4.25 (m, 4'-H), 6.52 (m, 1'-H), 7.74 (s, NH _2), 8 , 06 (s, 3-H), 8,24 (s, 6-H).

b) 110 mg (0.2 mmol) of the product were stirred with 10 ml of 80% acetic acid at room temperature for 20 minutes. After chromatography (silica gel, dichloromethane / methanol 9: 1), the desired product was obtained in crystalline form. Subsequent recrystallization from isopropanol / cyclohexane gave 40 mg (85%) of the product as a decomposed solid.

UV (ΜβΟΗ): λ ™ _χ = 260.275nm (ε = 9000.10200) C ₁₀ H ₁₃ N ₅ O ₂ (235.25) Calcd: C 51.06 H 5.57 N 29.77 V: C 50, 96 H 5.65 N 29.80

¹³ C NMR ([De] DMSO): δ = 133 (C-8), 100.3 (C-5), 158.1 (C-6), 156.1 (C-2), 153.6 (C-4), 84.4 (CV), 30.4 (C-2 '), 27.4 (C-3 ¹ ), 81.7 (C-4'), 64.3 (C-5 ')

TLC (silica gel, CH ₂ Cl ₂ / methanol, 9: 1 ,: R, = 0.4. UV (MrOH): A _max = 260.275nm (c = 9000.10200).

¹ H-NMR ((D ₆ ) DMSO): δ = 2.11 (m, 3'-H), 2.40 (m, 2'-H), 3.3 P (m, VH), 4.08 (m, 4'-H), 4.75 (m, 5'-OH), 6.45 (m, Γ-Η), 7.75 (s, NH ₂ ), 8.14 (s, 3-H) , 8,18 (s, 6-H).

-12- 269 8B4

Example 18

a) 4.6 · ΟΙοΗΙθΓ-1 · (2 '· αθ8θχγ · 3', 5'-αΙ ^ · | ^ οΙυογΙ · β · 0 · βΓνΙηΓθ · ρβΜο-ΙυΓβηο $ νΙ) · 1Η · ρνΗθΙο [3 _ι 2- ο] ρνΓΜΙη

A solution of 300mg (1.6mmol) of 4,6-dichloro-i H-pyrrolo (3,2-c) pyridine in dry acetonitrile (75ml) containing 450mg

(8.0 mmol) of potassium hydroxide and 30 mg (0.1 mmol) of tris (2-U-methoxyethoxy) ethyl] arnine was stirred at room temperature under nitrogen for 30 minutes. With stirring, 625 mg (1.6 mmol) of α-chloro-2-deoxy-3,5-di-O-p-toluoyl-D-erythro-pentofuranose were added and the mixture was stirred for a further 15 minutes. Insoluble material was filtered off. The filtrate was concentrated in vacuo. The oily residue was chromatographed on silica gel (17 x 4 cm column, eluent dichloromethane-ethyl acetate [97: 3]). 762 mg (90%) of the colorless anoropenic product were obtained.

¹ H-NMR (Me ₂ SO-d,): δ = 2.37 and 2.41 (2s, 2CH ₃ ), 2.77 (m, H-2's),? .S Im, H-2 ') , 4.57 (m, H-4 ¹ , H-5 '), 5.68 (m, H-3'), 6.66 (pt.

H-1 '), 6.71 (d, J = 3.5Hz, H-3), 8.00 (s, H-7),

¹³ C-NMR (MejSO-d _e ): δ = 36.8 (C-2 '), 64.2 (C-5'l, 74.9 (C-3 ¹ ), 81.7 (CT), 85.6 (C-4 '), 102.0 (C-3), 106.1 (C-7), 123.1 (C-3a), 129.7 (C-2), 140.0 ( C-6), 140.6 (C-4), 142.4 (C-7a).

b) 4,6-dichloro-1 • (2'-deoxy-β-D-erythro-pentofuranosyl. ') - 1 H -pyrrolo [3,2-c] pyridine

500 mg (0.93 mmol) of the compound from Example 18a) were dissolved in 30 ml of methanolic ammonia and stirred at 50 ° C for 12 hours. The solution was concentrated to dryness, the solid residue adsorbed on silica gel 60 H (2g) and applied to a silica gel column (14 x 4cm, eluent chloroform / methanol [9:11]. The main faction became the

Product isolated as a colorless oil which crystallized from aqueous ethanol in colorless needles. Yield: 101 mg (72%), mp 18O ⁰ C

¹ H-NMR (Me ₂ SO-d "): δ = 2.28 (m, H-2's), 2.43 (m, H-2'a), 3.56 (m, H-5 ') , 3.85 (m, H-4 '), 4.38 (m, H-3'), 5.02 (t, J = 5.2Hz, 5'-OH), 5.34 (d, J = 4 , 1 Hz, 3'-OH) ₁ 6.42 (pt, HV), 6.67 (d, J = 3.4Hz, H-3), 7.89 (d, J = 3.4Hz, H ' -2), 7.96 (s, H-7).

¹³ C-NMR (Me ₂ SO-d _e ): δ = 40.6 (C-2 '), 61.5 (C-5'l, 70.5 (C-3 ¹ ), 85.5 (CV ), 87.6 (C-4'l, 101.3 (C-3), 106.1 (C-7), 123.1 (C-3a), 129.7 (C-2), 139, 7 (C-6), 140.4 (C-4), 142.0 (C-7a).

c) 4-Amino-6-chloro-1- (2'-deoxy-β-D-erythro-pentofuranosyl) -1H-pyrrolo [3,2-c] pyridine

460 mg (1.52 mmol) of the compound from Example 18 b) were dissolved in 6 ml of dry hydrazine and heated to 80 ° C for 60 minutes. The hydrazine was removed in vacuo and the oily residue was concentrated twice with 10 ml of ethanol. The residue was dissolved in 40 ml of aqueous ethanol. Then, 2 g of Raney nickel catalyst was added and the mixture was heated to boiling with stirring for two hours. The catalyst was filtered off and washed thoroughly with fresh aqueous ethanol. The filtrate was concentrated to dryness, the residue dissolved in methanol, adsorbed on 2 g of silica gel and the solvent removed in vacuo. This silica gel was suspended in chloroform / methanol (9: 1) and applied to a silica gel column (6 × 3 cm). Elution with chloroform / methanol (9: 1) gave a colorless syrup from which, by crystallization from methanol, the product could be obtained in small, colorless crystals of melting point 232 ° C.

Yield: 207 mg, 48%

TLC (chloroform / methanol [9: 1]): R _r = 0.2; UV (methanol) A _mix = 277nm (ε = 14800), 285nm (ε = 13800); ¹ H-NMR (Me ₂ SO-d _a ): δ = 2.20 (m, H-2'm), 2.40 (m, H-2'a), 3.51 (m, H-5 '), 3.78 (m, H-4'), 4.32 (m, H-3 '), 4.89 (t, J = 5Hz, 5'-OH), 5.26 (d, J = 4Hz, 3'-OH), 6.19 (pt. HV), 6.55 (s, NH ₂ ), 6.64 (d, J = 3Hz, H-3), 6.83 (s, H -7), 7.36 (d, J = 3Hz, H-2).

¹³ C-NMR (Me ₂ SO-d ₈ ): δ = 40 (C-2 '), 61.8 (C-5 ¹ ), 70.6 (C-3'), 64.7 (CV), 87.2 (C-4 '), 95.1 (C-7), 101.6 (C-3), 109.6 (C-3a), 123.5 (C-2), 141.0 ( C-6), 141.4 (C-7a), 152.9 (C-4)

C ₁₂ H ₁₄ CIN ₃ O ₃ Ber. C50.80 H4.97 N 14.81 Cl 12.50% Gef. C 50.91 H 5.05 N 14.75 Cl 12.53%

d) 4-Amino-1- (2'-deoxy-β-D-erythro-pentofuranosyl) -1H-pyrrolo [3,2-c] pyridine

A solution of 200 mg (0.7 mmol) of the compound of Example 18c) in methanol (30 ml) containing 0.4 ml of methanol saturated with ammonia was dissolved in the presence of palladium-charcoal (50 mg, 10% Pd) at room temperature Hydrogenated for hours. The catalyst was filtered off and the solvent removed in vacuo. ([7: 3: 2] Column 4 x 4cm, eluent chloroform / methanol / triethylamine) Purification by flash chromatography and crystallization from methanol gave 70 mg (40%) of the product (as colorless crystals of melting point 205 ⁰ C. TLC eluent chloroform / methanol / Triethylamine [7: 3: 2] v / v / v): R, = 0.4; UV (methanol) A _n ,., = 271 nm (ε = 12800); ¹ H-NMR (Me ₂ SO-d "): δ = 2.20 (m, H-2'b), 2.42 (m, H-2'a), 3.51 (m, H-5 '), 3.80 (m, K-4'), 4.32 (m, H-3 '), 4.91 (m, 5'-OH), 5.32 (m, 3'-OH) , 6,08 (s, NH ₂ ), 6,23 (pt H-1 '), 6,65 (d, J = 3Hz, H-3), 6,75 (d, J = 6Hz, H- 7), 7.35 (d, J = 3Hz, H-2), 7.55 (d, J = 6Hz, H-6).

¹³ C-NMR (MejSO-de): δ = 39.8 (C-2 '), 62.0 (C-5'), 70.8 (C-3 '), 84.5 (CV), 87 , 1 (C-4 ¹ ), 96.9 (C-7), 101.5 (C-3), 110.7 (C-3a), 122.5 (C-2), 139.7 (C -6), 140.0 (C-7a), 153.7 (C-4)

Ci ₂ H ₁₅ N ₃ O ₃ Ber. C 57.82 H 6.07 N 16.86% Gef. C 57.97 H 6.12 N 16.74%

Example 19

a) 6-Chloro-1- (2'-dosoxy-.beta.-D-erythro-pentofuranosyl) -1H-pyrrolo [3] ic] pyridin-4one. A solution of 400 mg (1.32 mmol) of the compound from Example 18b) in 2 N aqueous sodium hydroxide solution (with small amounts of 1,4-dioxane) was heated to boiling for 30 hours. The reaction mixture was neutralized with 2N hydrochloric acid, filtered and then loaded onto an Amberlite XAD-4 column (17x2 cm). Inorganic salts were removed by washing with water. The product was aluiert with methanol. Crystallization from water gave 158 mg (42%) of colorless crystals. Melting point 242-243 ⁰ C. TLC (chloroform / methanol [8: 2]):

R, = 0.5 UV (methanol): X _mjx = 270 nm (ε = 11100), 292 nm (ε = 9300); ¹ H-NMR (Me ₂ SOd ₆ ): δ = 2.22 (m, H-2'b), 2.38 (m, H-2'a), 3.53 (m, H-5 ') , 3.80 (m, H-4 '), 4.33 (m, H-3'), 4.96 (m, 5'-OH), 5.29 (m, 3'-OH), 6 , 22 (pt HV), 6.54 (d, J = 3.3 Hz, H-3), 6.96 (s, H-7), 7.38 (d, J = 3.3Hz, H -2), 11.81 (NH, NH). ¹³ C-NMR (Me ₂ SO-d ₆ ): 40.5 (C-2 '), 61.7 (C-5'), 70.6 (C-3 '), 85.0 (CV), 87.4 (C-4 '), 94.9 (C-7), 104.1 (C-3), 114.0 (C-3a), 123.2 (C-2), 129.1 ( C-6), 139.2 (C-7a), 158.7 (C-4). C ₁₂ H ₁₃ CIN ₂ O ₄ Ber. C 50.63 H 4,60 N 9,84 Cl 12,45 Got. C 50.79 H 4.74 N 9.80 Cl 12.69

-13- 269 8ii4

b) 1- (2'-deoxy-β-D-erythro-pentofuranosyl) -1H-pyrrolo [3.2-c] pyridin-4-one

A solution of 100 mg (0.35 mmol) of the compound of Example 19a) in methanol (15 ml) was mixed with 0.5 ml of 25% aqueous ammonia and in the presence of palladium / animal charcoal (10% Pd, 15 mg) for 3 hours at room temperature hydrogenated. The catalyst was filtered off and the filtrate was concentrated to dryness. The solid residue was crystallized from water. This gave 51 mg (58%) of product of melting point 147-148 ⁰ C. TLC (eluant chloroform / methanol (8: 2]):

R (= 0.3; -UV (methanol): λ ™, = 264 nm (e = 11700), 282 nm (sh, ε = 8000), 295 nm (sh, e = 5100), - ¹ H-NMR (Me ₂ SO-d ₆ ): δ = 2.22 (m, H-2's), 2.40 (m, H-2's), 3.52 (m, H-5 '), 3.81 (m, H) 4 ¹ ), 4.32 (m, H-3's), 4.93 (t, J = 5.4 Hz, 5'-OH), 5.32 (d, J = 4.3 Hz, 3'- OH), 6.21 (pt H-1 '', 6.54 (d, J = 3Hz, H-3), 6.62 (d, J = 7Hz, H-7), 7.03 ( d, J = 7Hz, H-6), 7.34 (d, J = 3Hz, H-2), 10.87 (br. NH).

C-NMR (Me ₂ SO-dj): δ = 40 (C-2 ^f , superimposed on solvent signals), 61.8 (C-5 ¹ ), 70.7 (C-3 '), 84, 8 (C-1 '), 87.4 (C-4'), 93.8 (C-7), 104.6 (C-3), 115.9 (C-3 a), 122.0 ( C-2), 127.8 (C-6), 139.0 (C-7a), 159.6 (C-4), C ₁₃ H ₁₄ N ₂ O ₄ calc. C 59.08 H 6, 10 N 10.60% Gef. C 59.09 H 6.07 N 10.65%

Example 20

a) 1- (2'-deoxy-β-D-erythro-pentofuranosyl) -4,6-dichloro-5'-O- (4,4'-dimethoxytrityl) -1H-pyrrolo [3.2-c] pyridine

500 mg (1.65 mmol) of the compound from Example 18b) were concentrated to dryness with 10 ml of pyridine. The material was dissolved in 10 ml of dry pyridine; 0.7 ml (4.1 mmol) Hünigs base and 690 mg (2.0 mmol) 4,4'-dimethoxytrityl chloride were added. The solution was stirred for one hour at room temperature. After addition of 75 ml of 5% aqueous sodium bicarbonate solution was extracted with dichloromethane (2 x 75 ml). The combined organic phases were dried over sodium sulfate. The sodium sulfate was filtered off and the filtrate was concentrated. The residue was applied to a silica gel column (30 × 3 cm, eluent dichloromethane and dichlorometane / acetone [99: 1]). The product was recovered from the main fraction as a yellowish amorphous mass. The product was dissolved in ether and precipitated with η-hexane. Yield: 740 mg (74%).

¹ H-NMR (Me ₂ SO-d _e ): δ = 2.39 (m, H-2'b), 2.64 (m, H-2'a), 3.09 (m, H-5 '), 3.72 (s, 2OCH ₃ ), 3.96 (m, H-4'), 4.42 (m, H-3 '), 5.41 (d, J = 4.8Hz, 3 '-OH), 6.47 (pt, H-1'), 6.65 (d, J = 3.5Hz, H-3), 6.76-7.27 (aromat H), 7.76 (d, J = 3.5Hz, H-2), 7.89 (s, H-7), ¹³ C-NMR (Me ₂ SO-d _e ): δ = 40 (C-2 'superimposed on solvent signals ) 55.1 (2OCH ₃ ), 63.6 (C-5 '), 70.05 (C-3'), 85.0, 85.5, 85.5 (CV, C-4 ', OCDMT) , 101.3 (C-3), 106.2 (C-7), 123.2 (C-3a), 129.1 (C-2), 139.8 (C-6), 140.5. '"M), 142.3 (C-7a). C ₃₃ H ₃₀ CI ₂ N ₂ O ₅ Calculated. C 65.46 H 4.99 Cl 11.71 N 4.63%. Found: C 65.47 H 5.09 Cl 11.78 N 4.56%

b) 1- (2'-deoxy-β-D-ergothipentofuranosyl) -4,6-dichloro-5'-O- (4,4'-dimethoxytrltyl) -3'-O-phenoxythiocarbonyl-1H-pyrrolo [ 3,2-c] pyridine

300 mg (0.5 mmol) of the compound from example 20a) were dissolved in dry acetonitrile (11 ml). 350 mg (2.8 mmol) of 4-dimethylaminopyridine and 150 μΙ (1.1 mmol) of phenyl chlorothiocarbonate were added and the solution was stirred at room temperature for 16 hours. Subsequently, the reaction mixture was concentrated to dryness in vacuo. The residue was chromatographed on silica gel (eluent dichloromethane). The colorless product was isolated from the main fraction (310 mg, 84%).

¹ H-NMR (Me ₂ SO-d ₆ ): δ = 2.92 (m, H-2'a, b), 3.35 (m, H-5 '), 3.72 (s, 2OCH ₃ ), 4.43 (m, H-4 '), 5.89 (m, H-3'), 8.61 (pt. HV) .6.71 (d, J = 3.5Hz, H-3 ), 6.81-7.52 (aromatics H), 7.76 (d, J = 3.5Hz, H-2), 8.01 (s, H-7).

¹³ C-NMR (Me ₂ SO-d ₆ ): δ = 37.0 (C-2 '), 55.1 (2OCH ₃ ), 63.8 (C-5'), 83.0, 84.2 , 85.6, 86.0 (C-1 ', C-3', C-4 ', OCDMT) 10', 8 (C-3), 106.3 (C-7). 123.1 (C-3a), 128.9 (C-2), 140.1 (C-6), 140.6 (C-4), 142.4 iC-7a), 193.8 (C = S). C ₄₀ H ₃₄ Cl ₂ N ₂ OeS Ber. C 64.78 H 4.62 Cl 9.55 N 3.77 S 4.32% Gef. C 64.66 H 4.59 Cl 9.65 N 3.70 S 4.40%

c) 4,6-dichloro-1- (2 ', 3'-dideoxy-β-D-glycero-pentofuranosyl) -5'-O- (4,4'-dimethoxytrityl) -1H-pyrrolo [3,2- c] pyridine

170 mg (0.23 mmol) of the compound from Example 20b) and 15 mg (0.1 mmol) of 2,2'-azobis (2-methyl) propionitrile were dissolved in 10 ml of dry toluene under an argon atmosphere. With stirring, 140 μΙ (0.51 mmol) of tri-n-butylstannane were added and stirred at 80 ° C for 3 hours. The solvent was removed in vacuo and the residue chromatographed on silica gel (eluent dichloromethane). From the main fraction, 115 mg (85%) of the product was isolated. ¹ H-NMR (Me ₂ SO-d ₆ ): δ = 2.05 (H-3 '), 2.50 (H-2 ¹ , superimposed on signals from the solvent), 2.90-3.15 (m , H-5 '), 4.25 (m, H-4'), 6.38 (m, HV), 6.63 (d, J = 3.4Hz, H-3), 6.69-7 , 30 (aromatic H). 7.79 (d, J = 3.4Hz, H-2), 7.89 (s, H-7).

d) 2,6-dichloro-3,7-didezaza-2 ', 3'-dideoxy-9-β-D-ribofuranosyl-purine

The dimethoxytrityl protecting group of the compound from Betspiel 20c) was removed anale.τ to Example 24f).

e) 6-Amino-3,7-didesza, T, 3'-dideoxy-9-β-D-ribofuranosyl-purine

The compound of Example 20d) was treated with hydrazine and subsequently reduced with Raney nickel as described in Example 18c). This gave the compound described in Example 1 (D).

f) 3,7-didesaza-2 ', 3'-did3soxy-9-β-D-ribofuranosyl-purine

The compound from Example 20d) was hydrogenated with Pd / animal charcoal / hydrogen analogously to Example 24g). The compound already described in Example 1 A) was obtained.

g) 3,7-didesaza-2 ', 3'-dideoxy-9-.beta.-D-ribofuranos /! -purlin-6-one

The compound from Example 20d) was treated with sodium hydroxide solution as described in Example 19a) and then hydrogenated as described in Example 19b). This gave the compound already described in Example 1 E).

Example 21

2-amino- (2 ', i'-dide8OXY-ß-D-glycero-pentofurenosyl) -1H-pyrazolo [3,4-d] pyrimidln-4-one

This compound was prepared analogously to Wej described in Example 17 via 2-amino- (2'-deoxy-9-.beta.-D-ribofuranosyl) -1H-pyrazolo (3,4-d] pyrimidin-4-one and Barton Deoxygenation of 2-amino'j- (2'-deoxy-3'-O-methoxythiocarbonyl-5'-toluoyl-ribofuranosyO-1H-pyrazolo [3,4-d] pyrimidine 4-one

CI ₀ H ₁₃ N ₅ O ₃ (251.25) Calc .: C47.81 H5.22 N 27.88% Found: C 48.01 H 5.30 N 27.83%

¹³ C-NMR (DMSO-d _e ): δ = 135.1 (C-3), 99.7 (C-3a), 157.9 (C-4), 155.3 (C-6), 154 , 5 (C-7a), 83.8 (CV), 30.3 (C-2 '), 27.3 (C-3'l, 8ϊ, 6 (C-4'), 64.3 '( C-5 ').

¹ H NMR: δ = 6.19 (dd, 1'-H, j = 6.9.3.5Hz), 2.06 (m, 3'-H)

Schmp.:221°C

Example 22

3,7-Dldesaza-2'-deoxy-9-β-D-ribofuranosyl-purine (2'-deoxy-3,7-dldesaza-nebular) The compound from Example 18b) was assayed on palladium / animal charcoal (10% Pd). hydrogenated in ammoniacal methanol. The product was purified by filtration of the catalyst and concentration of the filtrate in vacuo of inorganic salts by chromatography on Amberlite XAD (methanol / water) and crystallization from water. Schmp.:175-176°C

¹³ C-NMR ([D ₆ ] DMSO): δ = 126.9 (C-2), 101.7 (C-3), 125.5 (C-3a), 143.3 (C-4), 140.6 (C-6), 105.9 (C-7), 139.2 (C-7a), 84.6 (CV), 70.8 (C-3 '), 87.8 (C) 4 '), 61.9 (C-5 ·)

UV (0.1 N aq. HCl): A _mix = 224.274 nm C ₁₂ H ₁₄ N ₂ O ₃ Calc .: C 61.53 H 6.02 N 11.96% Found: C 61.55 H 6, 12N 12.02%

¹ H-NMR (L MSO-de): δ = 2.23 (m, 2'-Hb), 2.29 (m, 2'-Ha), 2.55 (m, 5'-H ₂ ), 3.85 (m, 4'-H), 4.38 (m, 3'-H), 4.99 (5'-OH), 5.37 (3'-OH), 6.42 (pt, I-H), 6.66 (d, J = 3Hz, 3-H), 7.62 (d, j = 6Hz, 7-H), 7.71 (d, j = 3Hz, 2-H) , 8.21 (d, j = 6Hz, 6-H), 8.23 (s, 4-H).

Example 23

a) 2-Chloro-e-n.sub.3-thoxy-3,7-didezaZe-2'-deoxy-9-.beta.-D-ribofuranosylpurene

The compound from example 18b) was heated for 40 hours in 1N methanolic sodium methanolate. The reaction product was purified on Amberlite XAD by hydrophobic chromatography (methanol / water). UV (methanol): A _n ,,,, = 271,280nm

C ₁₃ H ₁₅ CIN ₂ O ₄ Ber. C 52.27 H 5.06 Cl 11.87 N 9.38% Gef. C52.24 H5.14 Cl 12.05 N9.46%

b) 2-chloro-3,7-dideaza-2'-deoxy-9-.beta.-D-ribofuranosyl-purine-6-one

Heating the compound of Example 18 b) in 2N aqueous sodium hydroxide / 1,4-dioxane for 30 hours gave 2-chloro-3,7-dideaza-2'-deoxy-9-.beta.-D-ribofuranosyl-purin-6-one: UV (Methanol): A _n ,,,, = 262 nm Ci ₃ H ₁₆ N ₂ O ₄ Calc .: C59.08 H6.10 N 10.60% Found: C 59.09 H 6.07 N 10.65 %

¹ H-NMR ([D ₆ ] DMSO): δ = 2.22 (m, 2'-H ") 2.38 (m, 2'-H ₁ ), 3.53 (m, 5'-H ₂ 3.80 {m, 4'-H), 4.33 (m, 3'-H), 4.96 (5'-OH), 5.29 (3'-OH), 6.22 ( pt, 1'-H), 6.54 (d, j = 3Hz, 3-H), 6.96 (s, 7-H), 7.38 (d, j = 3Hz, 2-H) , 11.81 (NH).

Example 24

a) 4-Chloro-7- (2'-deoxy-3,5-dl-O- (p-toluoyl) -β-D-erythropentofuranosyl) -7H-pyrrolo [2,3-d] pyridine> idine 1 g ( 17.8 mmol) of powdered potassium hydroxide was added at room temperature in 60 ml of dry acetonitrile. With stirring, 100 μΙ (0.31 mmol) of tris [2- (2-methoxyethoxy) ethyl] amine was added. After 5 minutes, 1.23 g (8.01 mmol) of 4-chloro-7H-pyrrolo [2,3-d] pyrimidine was dissolved in the reaction mixture and stirred for an additional 5 minutes. Then, a-chloro-2-deoxy-3,5-di-Opo-toluoyl-β-D-erythro-pento-furanose was added. After stirring for 15 minutes, insoluble material was removed by filtration. The filtrate was concentrated to dryness in vacuo and the residue was chromatographed on a silica gel column (5 x 7cm, chloroform). Concentration of the eluate in vacuo gave 3.26 g (81%) of product which crystallized from ethanol in colorless needles (mp 120 ° C).

 Other variants of the manufacturing process:

(I) Solid-liquid glycosylation in the absence of a catalyst: The reaction was carried out as described above but without catalyst. After working up, 2.82 g (70%) of the product were obtained.

(II) By liquid-liquid phase transfer glycosylation:

500mg (3.26mmol) of 4-chloro-7H-pyrrolo [2,3-d] pyrimidine was dissolved in 20ml of dichloromethane. 9 ml of 50% aqueous sodium hydroxide solution was added. After adding 10 mg (0.03 mmol) of tetrabutylammonium hydrogensulfate, the solution was stirred with a vibromixer for one minute. Next, 1 .4n (3.61 mmol) of the above-described halogenose was added and mixed for a further three minutes. Thereafter, the phases were found. The aqueous phase was extracted twice with 25 ml of dichloromethane each time. The combined organic Phast-T were dried over sodium sulfate. The filter was evaporated to dryness. The residue was chromatographed on silica gel (column 5 × 5 cm, chloroform). Isolation of main fraction material and crystallization from ethanol erpab 1.04g (63%) product. Melting point: 118 ^° C.

TLC (cyclohexane / ethyl acetate 3: 2): Rf = 0.7. UV (MeOH): A _n ,,, = 240 nm (log [e] = 4.48).

'H-NMR (DMSO-de): δ = 2.37, 2.40 (s, 2CH ₃ ); 2.77 (m, 2'-H "); 3.18 (m, 2'-Ha); 4.60 (m, 4'-H and 5'-H); 5.77 (m, 3'-H); 6.75 (d, ο = 3.7Hz, 5-H); 6.78 (m, VH); 7, 34, 7, 91 (m, 8 aromatic, H and β-Η); 8.65 (s, 2-H).

b) 4-Chloro-7- (2'-deoxy-9-.beta.-D-erythro-pentofuranosyl) -7H-pyrrolo [2 ,: J] pyrimidine

2.4 g (4.7 mmol) of the compound from Example 24a) were stirred in 10 μl of ammonia-saturated methanol for 24 hours at room temperature. The solution was concentrated to dryness, the residue was adsorbed on silica gel 6OHMOg) and applied to a silica gel column (4 × 10 cm, chloroform / methanol, 95: 5). From the main fraction, the product was isolated as a colorless solid which crystallized from ethyl acetate in colorless needles. Yield: 1.07 g (84%). Melting point 162 ° C. CC (chloroform / methanol, 9: 1): R _f = 0, v.

UV (ΜβΟΗ): λη ,, _χ = 273nm (Löge = 3.69). ¹ H-NMH (DMSO-d _e ): δ = 2.28 (m, 2'-Hb); 2.58 (m, 2'-Ha); 3.57 (m, 5'-H); 3.87 (m, 4'-H); 4.40 (m, 3'-H); 5.00 (t, J = 5.4Hz, 5'-OH), 5.35 (d, J = 4.2Hz, 3'-OH); 6.66 (m, Γ-Η); 6.72 (d, J = 3.8Hz, 5-H); 7.99 (d, J = 3.8Hz, 6-H); 8.66 (s, 2-H).

c) 4-Chloro-7- (2'-deoxy-β-D-orythro-pentofuranosyl-5'-O- (4,4'-dimethoxytrltyl) -7H-pyrrolo [2,3-d] pyrimididine

1 g (3.7 mmol) of the compound from Example 24 b) were dried by concentration with 10 ml of dry pyridine. The material was dissolved in dry pyridine (20 ml). 2rr (11.7 mmol) Hünigs base and 2g (5.9 mmol) 4,4'-dimethoxytrityl chloride were added. The solution was stirred at room temperature for three hours. After addition of 80 ml of 5% strength aqueous sodium carbonate solution, the solution was extracted with 3 × 100 ml of dichloromethane. The combined organic phases were dried over sodium sulfate. After filtration, the filtrate was concentrated in vacuo. The residue was purified by column chromatography (silica gel, eluent dichloromethane and dichloromethane / ethyl acetate, 9: 1). Isolation of the major fraction material, dissolution in ether and petroleum ether cases gave 1.66 g (78%) of yellowish amorphous substance.

C ₃₂ H ₃₀ N ₃ O ₅ Cl (572.1) Ber. C 67.19 H 5.29 Cl 6.20 N 7.35% Gef. C 67.03 H 5.47 Cl 6.19 N 7.29%

TLC (CH ₂ Cl ₂ / acetone, 9: 1): R, = 0.3.

UV (MeOH): A _011x = 274nm (logle] = 3.85).

¹ H-NMR (DMS0-d _s): δ = 2.36 (m, 2'-H _b); 2.70 (m, 2'-H ₁ ); 3.72 (see OCH ₃ ); 3.18 (d, J = 4.5Hz, 5'-H); 3.99 (m, 4'-H); 4.45 (m, 3'-H), 5.42 (d, J = 4.6Hz, 3'-OH); 6.65 (m, 1'-H); 6.69 (d, J = 3.7Hz, 5-H); 7.81 (d, J = 3.7Hz, 6-H), 8.64 (s, 2-H).

d) 4-Chloro-7 (2'-deoxy-β-D-erythro-pentofuranosyl) -5'-O- (4,4'-dimethoxytrityl) -3'-O-phenoxythocarbonyl-7H-pyrrolo [2,3 -d -] - pyrimidine

1 g (1.7 mmol) of the compound of Example 24c) was dissolved in 30 ml of dry acetonitrile. 500 mg (4.1 mmol) of 4-dimethylaminopyridine and 400 μΙ (2.9 mmol) of phenyl chlorothiocarbonate were added and the solution was stirred at room temperature for 16 hours. The reaction mixture was then concentrated to dryness in vacuo and the residue was applied to a silica gel column (3 × 15 cm, dichloromethane). From the main fraction 950 mg (76%) of colorless amorphous product were isolated.

C ₃₉ H ₃₄ CIN ₃ O ₆ S (708.2) Ber. C 66.14 H 4.84 Cl 5.01 N 5.93 S 4.53 Gef. C 66.22 H 4.9 'Cl 5.12 N 5.93 S 4.46

TLC (CH ₂ C! ₂ / acetone 95: 5) R, = 0.8.

UV (MeOH): A _mjx = 274 nm (log [e] = 3.87).

¹ H-NMR (DMSO-d _e ): δ = 2.84 (m, 2'- _Hb ); 3.21 (m, 2'-H ₁ ); 3.37 (m, 5'-H); 4.46 (in, 4'-H); 5.92 (m, 3'-H); 6.70 (m, 1'-H); 6.76 (d, J = 3.8Hz, 5-H); 7.85 (d, J = 3.8Hz, 6-H); 8.61 (s, 2-H).

e) 4-chloro-7- (2 ', 3'-dideoxy-p-D-glycero-pentofuranosyl) -5'-O- (4,4'-dimethoxytitryl) -7H-pyirolo [2,3-d] pyrimidine

800 mg (1.1 mmol) of the compound of Example 24d) and 40 mg (0.2 mmol) of 2,2'-azobis (2-mothyl) propionitrile were dissolved in 40 ml of dry toluene under argon atmosphere. 600μI (2.2 mmol) of tri-n-butyl stannane was added with stirring and the reaction was continued at 75 ⁰ C for two hours. The solvent was removed in vacuo and the residue was chromatographed on silica gel (column 15 x 3 cm, dichloromethane / ethyl acetate 95: 5). From the main fraction, 470 mg (75%) of the product were obtained.

C ₃₂ H ₃₀ CIN ₃ O ₄ (556.1) Ber. C 69.12 H 5.44 Cl 6.38 N 7.56% Gef. C 69.07 H 5.53 Cl 6.33 N 7.58%

TLC (CH ₂ Cl ₂ / acetone 95: 5): R _; = 0 /.

UV (MeOH): X _m "= 273 nm (logU) = 3.78).

¹ H-NMR (DMSO-de): δ = 2.08 (m, 3'-H); 2.10 (m, 2'-H "); 2.43 (m, 2'-H ₁ ); 3.11 (d, J = 4.4Hz, 5'-H), 3.71 (s, OCH ₃ ); 4.27 (m, 4'-H); 6.55 (dd, J = 3.6 and 6, SHz, 1'-H); 6.64 (d, J = 3.7Hz, 5-H); 7.83 (d, J = 3.7Hz, 6-H); 8.67 (s, 2-H).

f) 4-Chloro-7- (2 ', 3'-diaesoxy-β-D-glycero-pentofuranosyl-7H-pyrrolo [2,3-d] -pyrimidine

400 mg (0.7 mmol) of the compound from Example 24e) were dissolved in 15 ml of 80% aqueous acetic acid and stirred at room temperature for 30 minutes. The solvent was removed in vacuo and traces of acetic acid removed by concentration with water. The residue was purified by column chromatography (dichloromethane and dichloromethane / methanol, 98: 2). From the main fraction, 120 mg (67%) of product was crystallized from ethyl acetate as colorless needles after crystallization. Melting point 9O ⁰ C.

C ₁₁ H ₁₂ CIN ₃ O ₂ (253.7) Ber. C52.08 H4,77 CI13.98 N 16,56 Gef. C 52,20 H 4,81 Cl 14,04 N 16,54

TLC (CH ₂ Cl ₂ / MeOH 95: 5): R, = 0.5

UV (MeOH): K _n ^ = 274nm (logle] = 3.65).

¹ H-NMR (DMSO ^ _O ) IO = 2.04 (m, 3'-H); 2.28 in, 2'- _Hb ); 2.46 (m, 2'-H ₁ ); 3.57 (m, 5'-H); 4.11 (m, 4'-H); 4.95 (t, J = 5.5Hz, 5'-OH); 6.52 (dd, J = 3.8 and 6.9Hz, 1'-H): 6.69 (d, J = 3.8Hz, 5-H); 8.01 (d, J = 3.8Hz, 6-H); 8.64 (s, 2-H).

g) 7- (2 ', 3'-Dideoxy-β-D-gero-pentofuranoiyl) -7H-pyrrolo [2,3-d] pyrrole

A solution of 200 mg (0.8 mmol) of the compound from Example 24f) was added to 20 ml of methanol, to which 0.5 ml (6.6 mmol) of aqueous ammonia had been added, with palladium on animal charcoal (40 mg, 10% Pd) in a white matter atmosphere at room temperature for three hours. The catalyst was filtered off and the solvent removed in vacuo. The residue was dissolved in water and chromatographed on an Amberlite XAD-4 column (1st eluent water, 2nd eluent water / methanol [8: 2]). Isolation of the main zone material yielded 130 mg (75%) of the colorless product in needles. Mp 131 ⁰ C. C _n H ₁₃ O ₂ N ₃ (219.2) Calcd. C 60.26 H 5.98 N 19.17% Gef. C 60.19 H 5.97 N 19.18%

TLC (CHjClj / MeOH 9: 1): R, = 0.6 UV (MeOH): A _n ,,,, = 270 nm (log) = 3.56).

'H NMR (DMSO-d'): δ = 2.06 (m, 3'-H); 2.27 (m, 2'- _Hb ); 2.42 (m, 2'-Ho); 3.55 (m, 5'-H); 4.09 (m, 4'-H); 4.93 (t, J = 5.5Hz, 5'-OH); 6.54 (dd, J = 4.3 and 6.9Hz, 1'-H); 6.67 (d, J = 3.7Hz, 5-H); 7.86 (d, J = 3.7Hz, 6-H); 8.79 (s, 4-H); 9.01 (s, 2-H).

h) 4-Amino-7- (2 ', 3'-dideoxy-β-D-flipercero-pentofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine (2', 3'-dideoxytubercidin) 200 mg (0 , 8 mmol) of the compound from Example 24f) were stirred in 60 ml of 25% aqueous ammonia for 15 hours or 100 ⁰ C under pressure in a steel bomb. Then the solvent was removed in vacuo and the residue was dissolved in 200 ml of water. This solution was purified on Dowex 1x2 (ear-shaped). The column was washed with water and the product was eluted with water / methanol (9: 1). From the home zone, 120 mg (65%) product was recovered. TLC (CH ₂ Cl ₂ / MeOH 9: 1): R, = 0.3

¹ H-NMR (DMSO-de): δ = 2.03 (m, 3'-H); 2.22 (m, 2'-H ₁ ); 2.33 (m, 2'- _Hb ); 3.53 (m, 5'-H); 4.04 (m, 4'-H); 4.99 (m, 5'-OH); 6.35 (m, VH); 6.51 (d, J - 3.6Hz, 5-H); 7.00 (s, NH ₂ ); 7.34 (, J = 3.6Hz, 6-H); 8.04 (s, 2-H).

i) 7- (2 ', 3'-dideoxy-β-D-glycero-pentofurano-8-yl) -4-methoxy-7H-pyrrolo [2,3-d] pyrimidine 170 mg (0.7 mmol) of the compound from Example 24f) dissolved in 5 ml of 1M methanolic methanolate solution and stirred at room temperature for four hours. The solution was neutralized with 80% acetic acid, concentrated in vacuo and the residue was applied to a silica gel column (eluent dichloromethane / methanol, 98: 2). Isolation of the major zone yielded a colorless oil which crystallized upon storage in needles. Yield: 130mg (78%)

j) 7- (2 ', 3'-Dldesoxy-.beta.-D-Grycero-pentofuranosyl) -4 H -pyrrolo [2,3-dl-pyrimidin-4-one 200 mg (0.8 mmol) of the compound from Example 24f) was added in 10 ml of 2N sodium hydroxide solution and heated to boiling for five hours under reflux. The solution was neutralized with 80% acetic acid and the insoluble material removed by filtration. The filtrate was applied to an Amberlite XAD-4 column. The column was washed with 500 ml of water and the product was eluted with water / 2-propanol (9: 1). This gave 180 mg (80%) of product.

Example 25

1- (2 ', 3'-dideoxy-.beta.-D-glycero-pentafuranosyl) -1H-pyrezolo [3,4-d] pyrimldin-4-one

The product of Example 17d) was deaminated with adenosine deaminase from calf intestinal mucosa cells. The progress of the Reaction was te: 275nm monitored by UV spectroscopy. The reaction gives the product quantitatively in the form of colorless crystals.

M.p. 171 ⁰ C

UV (MeOH): A _mix = 251 nm (ε = 7700) TLC (silica gel, dichloromethane / methanol 9: 1): R ₍ = 0.5

¹³ C-NMR ([D ₆ I-DMSO]: δ = 135.2 (C-8), 106.1 (C-5), 107.3 (C-6), 148.4 (C-2) , 152.3 (C-4), 84.6 (CV), 30.7 (C-2 '), 27.3 (C-3 ¹ ), 82.2 (C-4'), 64.2 (C- ^5f ).

¹ N-NMR (ID ₆ I-DMSO): δ = 2.13 (m, 3'-H), 2.40 (m, 2'-H), 3.40 (m, 5'-H), 4.09 (m, 4'-H) 4.73 (m, 5'OH), 6.43 (m, VH), 8.11 (s, 3-H),

Example 26 2-Amino-7-deaza-2 ', 3'-dideoxy-9-β-D-ribofuranosyl-purin-6-one 5'-triphosphate

C ₁₁ H ₁₄ N ₄ O ₁₂ P ₃ Na ₃ (556.2) Calcd. P: 16.7 Gef. P: 16.4

UV (buffer, pH 7.0): A _m "= 259 nm (e = 13400)

³¹ P-NMR (D ₂ O): δ = -8.35 (d, Pv), -10.0 (d, Pa), -21.5 (t, P-β)

Example 27 2-Amino-3,7-didecaze-2'-deoxy-9-β-D-ribofuranosyl-purine-e-on-5'-triphosphate

C ₁₂ H ₁₆ N ₃ O ₁₃ P ₃ Na ₃ (555.2) Calcd. P: 16.75 Gef. P: 16.5

UV (pH 7.0 buffer): X _n ,,, = 272nm (e = 12400) Example 28

3,7-Dldesaza-2 ', 3'-dldesoxy-9-.beta.-D-ribofuranosyl-purine-5'-tri-phosphate C ₁₂ H ₁₄ N ₂ O ₁₁ P ₃ Na ₃ (524.1) Calcd. P: 17.7 Gef. P: 17.3

UY (buffer, pH 7.0): A _n ,,, = 224.274 nm

All the triphosphates listed in Examples 26 to 28 were prepared by phosphorylation of the corresponding Yoshikawa nucleosides (Tetrah. Lett. 50, 5065 (1967)) to the 5'-monophosphate and subsequent conversion into the 5'-triphosphate according to Hoard and Ott (J. Chem. Soc. 87, (1965) 1785).

Example 29 Antiviral activity

The stability of the N-glycosidic linkage of 2 ', 3'-dideoxy-nucleosides is associated with the antiviral activity.

The hydrolysis of the bond was examined at 25 ° C at three different concentrations of hydrochloric acid. For this purpose, the decrease in UV adsorption (E,) at 258 nm was measured. The rate-constant of the hydrolysis (k) and the half-lives (Va) were determined by the absorption / time curve from the equation K = l / t × In (E ₀ -E ₀₀ ) / (Et - E ₀₀ ). Thus, E _{o is the} absorption - ο and E _{00 is} the absorption after completion of the reaction.

Compared were 2 ', 3'-dideoxyadenosine (a) and 6-amino-8-aza-7-deaza-2', 3'-dideoxy-9-β-D-ribofuranosyl-purine (b)

Table 1

1NHCI 0.1NHCI 0.01NHCI

(a) ^T / 2 - 1.9 min 31.5 min

k - 0.363 mh ~ ¹ 0.022 min " ¹

(b) τ / 2 0.83 min 20.4 me. 280 min k 0.85 min " ¹ 0.033 min" ¹ 0.0025 min " ¹

Table 1 shows that compound (b) according to the invention is more than 10 times more stable and thus more antivirally more effective than (a).

Claims (1)

claims: in the X nitrogen or a methine group, W is nitrogen or the group CR ⁴ R ¹ , R ² , R ³ , R ⁴ , which may be the same or different, are hydrogen, halogen, lower alkyl, hydroxy, mercapto, lower alkylthio, lower alkyloxy, aralkyl, aralkyloxy, aryloxy or optionally mono- or di-substituted amino group, R ^{5 is} hydrogen or a hydroxy group, R ⁶ , R ^{7 are} each hydrogen or one of the two radicals R ⁶ and R ^{7 is} halogen, a cyano, an acid or an optionally mono- or di-substituted amino group, wherein one of the radicals R ³ and R ⁷ also represent a hydroxy group can, if X means a methine group, and also R ⁵ and R ⁷ together may represent another bond between C-2 'and C-3' Y represents hydrogen, a monophosphate, diphosphate or triphosphate group, as well as possible tautomers and salts and nucleic acids which contain one or more compounds of the formula I as building block, characterized in that compounds of the formula II, (ID ₁ in which X, W, R ¹ , R ² and R ^{3 have} the meaning given in claim 1, with a compound of formula III R> -0 (III) in the R ^{5 has} the meaning given above, R ⁶ ', R ⁷ ' are each hydrogen or one of both radicals R ⁶ 'and R ⁷ ' is an azido or a hydroxy group protected by an oxygen protecting group, R 'is an oxygen protecting group and
Z is a reactive group
to compounds of Formula IV R ' R'-0 HV)