New nucleoside and oligonucleotide analogues

 

The invention relates to nucleoside analogs of formula (1) in which R1represents H or a group protecting the hydroxyl, R2represents H, a group protecting the hydroxyl group of phosphoric acid, a protected group, phosphoric acid or a group of the formula P(R3R4in which R3and R4are the same or different and represent a hydroxyl group, a protected hydroxyl group, alkoxygroup, allylthiourea, cyanoacetylurea, amino group, substituted alkyl group; And represents alkylenes group containing from 1 to 4 carbon atoms, and a represents a substituted purine-9-ilen group or substituted 2-oxopyrimidine-1-ilen group containing at least one Deputy, selected from hydroxyl groups, protected hydroxyl groups, amino groups, protected amino groups, alkyl groups. The invention relates to oligonucleotide analogues comprising one or more structures of the formula (2), where a and b have the same meanings as in the formula (1). Oligonucleotide analogues according to the invention or their pharmaceutically acceptable salts can be used as drugs, possessing the">

The technical FIELD TO WHICH the INVENTION RELATES

The present invention relates to the field of organic chemistry, in particular to novel oligonucleotide analogs which are antisense activity or antigenic activity, possess excellent stability or exhibit excellent activity as the detecting agent (probe) for a specific gene or as a primer for starting amplification, and to new nucleoside analogues, which are intermediate compounds for their preparation.

The LEVEL of TECHNOLOGY

I believe that oligonucleotide analogues with excellent antisense or antigenic activity and stable in the body, may be useful in pharmaceutical preparations. In addition, oligonucleotide analogues with high ability to form a stable complex with DNA or mRNA, are useful as detection agents for specific genes or as primers for the initial amplification.

On the contrary, it is known that oligonucleotides of natural origin quickly disintegrate under the action of various nucleases, p is to provide sufficient sensitivity when used as detecting agents for specific genes or as a primer for starting amplification due to the limitations of their affinity with complementary sequences.

In order to overcome these disadvantages, have been obtained in different naturally occurring oligonucleotide analogues and attempts were made to their use as pharmaceuticals or detecting agents for specific genes. Known examples of such are not found in nature oligonucleotide analogs include oligonucleotides, in which the oxygen atom bound to the phosphorus atom phosphorodithioic communication, replaced by a sulfur atom; oligonucleotides in which the specified oxygen atom substituted methylene group; oligonucleotides in which the specified oxygen atom replaced by a boron atom, and oligonucleotides, in which the functional group of the sugar or the main functional group of the oligonucleotide is chemically modified. For example, the firm ISIS Corp. developed oligonucleotide timetogo type ISIS2922 (Vitravene) as a therapeutic agent for the treatment of cytomegalovirus retinitis person, and in the United States oligonucleotide ISIS2922 entered the free market.

However, the potential efficacy of antisense activity or antigenic activity of the above, not naturally occurring oligonucleotide analogues, namely the ability to education is cnyh effects due to non-specific binding to various proteins in the body, makes the actual development is not occurring oligonucleotide analogs with greater stability in the body that have a low probability of unwanted side effects and a high ability to form stable complementary circuits.

The INVENTION

The authors of the present invention over a long period of time spent in intensive study does not naturally occurring oligonucleotide analogues having excellent antisense or antigenic activity, excellent stability in the body and a low likelihood of unwanted side effects. In this study, the authors found that oligonucleotide analogs or nucleoside analogues having ether bond in these molecules, are useful as antisense or antigenic pharmaceutical preparations having excellent stability, the detecting agent (probe for a particular gene, a primer for starting amplification or as intermediate compounds for their production, and completed the present invention.

In the following the present invention will be described in detail.

New nucleoside analogues of the present izopet 2 are the same or different substituents and represent a hydrogen atom, a group protecting the hydroxyl group of phosphoric acid, a protected group, phosphoric acid or a group-P(R3R4[in which R3and R4are the same or different and represent a hydroxyl group, a protected hydroxyl group, mercaptopropyl, secure mercaptopropyl, amino group, alkoxygroup having from 1 to 4 carbon atoms, allylthiourea containing from 1 to 4 carbon atoms, cyanoacetylurea containing from 1 to 5 carbon atoms, or amino group, substituted alkyl group containing from 1 to 4 carbon atoms;

But alkylenes group having from 1 to 4 carbon atoms, and

Represents a purine-9-ilen group, 2-oxo-pyrimidine-1-ilen group or a substituted purine-9-ilen group or substituted 2-oxopyrimidine-1-ilen group with the Deputy, selected from the following alpha group,

or their salts.

Oligonucleotide analogues of the present invention are oligonucleotide analogs that have one, or two or more structures of the formula (2)

in which a represents sobomehin-1-ilen group, or substituted purine-9-ilen group, or substituted 2-oxopyrimidine-1-ilen group having a Deputy chosen from the following alpha group,

or their pharmacologically acceptable salts.

(alpha group):

hydroxyl group,

protected hydroxyl group,

alkoxygroup having from 1 to 4 carbon atoms,

mercaptopropyl,

protected mercaptopropyl,

allylthiourea having from 1 to 4 carbon atoms,

amino group,

protected amino group,

amino group, a substituted alkyl group having from 1 to 4 carbon atoms,

an alkyl group having from 1 to 4 carbon atoms, and

halogen atom.

"Allenova group having from 1 to 4 carbon atoms" group a in the above formula (1) or (2) may include methylene, ethylene, trimethylene and tetramethylene group, preferably a methylene group.

The protective group that protects the hydroxyl", R1and R2and "protected hydroxyl group, R3and R4or the alpha group in the above formula (1) or (2) refers to a protective group which can be chipped off by a chemical method such as hydrogenolysis, decomposition, hydrolysis, electrolysis and photolysis, or biological is kind of acyl group", such as alkylcarboxylic group, such as formyl, acetyl, propionyl, butyryloxy, isobutyryloxy, pentanoyl, pivaloyl, valerino, isovaleryl, octanoyl, nonanoyl, technology, 3-methylnonanoic, 8-methylnonanoic, 3-ethyloctanoic, 3,7 - dimethylacetanilide, undecanoyl, dodecanoyl, tridecanol, tetradecanol, pentadecanol, hexadecanol, 1-methylpentanol, 14-methylpentanol, 13,13-dimethylacetanilide, heptadecanoyl, 15-methylhexadecanoic, octadecanoyl, 1-methylheptadecyl, nanotechnology, eicosanol and heneicosanol, carboxypropanoyl alkylcarboxylic group, such as succinoyl, glutaryl and adipoyl, halogen(lower)alkylcarboxylic group such as chloroacetyl, dichloroacetyl, trichloroacetyl and TRIFLUOROACETYL, lower alkoxy(lower)alkylcarboxylic group, such as methoxyacetyl, and unsaturated alkylcarboxylic group, such as (E)-2-methyl-2-butanol;

"aromatic acyl group" such as arylcarbamoyl group, for example benzoyl, alpha naphtol and beta naphtol, halogenoalkanes group, such as 2-bromobenzoyl and 4-chlorobenzoyl, lower alkilirovanny and the group, for example, 4-anisoyl, carboxypropanoyl arylcarbamoyl group, such as 2-carboxybenzoyl, 3-carboxybenzoyl and 4-carboxybenzoyl, nitrated arylcarbamoyl group, for example 4-nitrobenzoyl and 2-nitrobenzoyl, lower alkoxycarbonylmethyl arylcarbamoyl group, such as 2-(methoxycarbonyl)benzoyl and anilinophenol arylcarbamoyl group, for example 4-phenylbenzyl;

"tetrahydropyranyloxy group or tetrahydropyranyl group, such as tetrahydropyran-2-ilen, 3-bromotetradecane-2-ilen, 4-methoxyacridine-4-ilen, tetrahydrothiopyran-2-ilen and 4-methoxytryptamine-4-ilen;

"tetrahydropyranyloxy group or tetrahydrofuranyl group" such as tetrahydrofuran-2-ilen and tetrahydrothiopyran-2-ilen;

"silyl group" such as tri(lower)alkylsilane group, for example trimethylsilyl, triethylsilyl, isopropylimidazole, tert-butyldimethylsilyl, methyldiisopropanolamine, methyldi-tert-Boticelli and triisopropylsilyl and three(lower)alkylsilane group, substituted by one or two aryl groups, for example diphenylmethylsilane, diphenylbutyric, diphenylethylene and phenyldimethylsilane;

"lower alkoxymethyl group, the tert-butoxymethyl;

"lower alkoxycarbonyl (lower)alkoxymethyl group" such as 2-methoxyethoxymethyl;

"halogen(lower)alkoxymethyl group" such as 2,2,2-trichloroacetyl and bis(2-chloroethoxy)methyl;

"lower alkoxycarbonyl ethyl group" such as 1-ethoxyethyl and 1-(isopropoxy)ethyl;

"halogenated ethyl group" such as 2,2,2-trichloroethyl;

"methyl group substituted by from 1 to 3 aryl groups such as benzyl, alpha-naphthylmethyl, beta-naphthylmethyl, diphenylmethyl, triphenylmethyl, alpha naphthylmethyl and 9-antimetal;

"methyl group substituted by from 1 to 3 aryl groups in which the specified aryl ring substituted lower alkyl, lower alkoxy, halogen atom or cyano", such as 4-methylbenzyl, 2,4,6-trimethylbenzyl, 3,4,5-trimethylbenzyl, 4-methoxybenzyl, 4-methoxyphenylalanine, 4,4'-dimethoxybiphenyl, 2-nitrobenzyl, 4-nitrobenzyl, 4-Chlorobenzyl, 4-bromobenzyl and 4-cyanobenzyl;

"lower alkoxycarbonyl group, such as methoxycarbonyl, etoxycarbonyl, tert-butoxycarbonyl and isobutoxide;

"lower alkoxycarbonyl group substituted by a halogen atom, or three(lower)alkylsilane group" such as 2,2,2-trichlorocyanuric xianbei, and

"aracelikarsaalyna group in which the specified aryl ring may be substituted by one or two lower alkoxy or nitro groups" such as benzyloxycarbonyl, 4-methoxybenzenesulfonyl, 3,4-dimethoxyphenylacetone, 2-nitrobenzeneboronic and 4-nitrobenzenesulfonyl.

"Protection of hydroxyl groups", R1and R2may respectively include "aliphatic acyl group", "aromatic acyl group", "methyl group substituted by from 1 to 3 aryl groups", "methyl group substituted by from 1 to 3 aryl groups in which the specified aryl ring substituted lower alkyl, lower alkoxy, halogen atom or cyano", or silyl group; more preferably acetyl group, benzoyloxy group, benzyl group, para-methoxybenzyloxy group, dimethoxytrityl group, monomethoxypolyethylene group or tert-butyldiphenylsilyl group.

The protective group of the protected hydroxyl group", R3and R4or the alpha group may preferably include aliphatic acyl group or aromatic acyl group", more preferably benzoyloxy group.

The protective group of the protected group Fosforit be chipped off by a chemical process, such as hydrogenolysis, hydrolysis, electrolysis and photolysis, and biological method such as hydrolysis in the human body, and such a protective group may include a "lower alkyl group" such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl and 2-ethylbutyl;

"sanirovano lower alkyl group" such as 2-cyanoethyl and 2-cyano-1,1-dimethylethyl;

"ethyl group, substituted silyl group" such as 2-methyldiphenylamine, 2-trimethylsilylmethyl and 2-triphenylsilanol;

"halogenated lower alkyl group" such as 2,2,2-trichloroethyl, 2,2,2-tribromoethyl, 2,2,2-triptorelin and 2,2,2-trichloro-1,1-dimethylethyl;

"lower alkenylphenol group, such as ethynyl, 1-propenyl, 2-propenyl, 1-methyl-2-propenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 2-ethyl-2-propenyl, 1-butenyl, 2-butenyl, 1-methyl-2-butenyl, 1-methyl-1-butenyl, 3-methyl-2-butenyl, 1-ethyl-2-butenyl, 3-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl, 1-ethyl-3-butenyl, 1-pentenyl, 2-pentenyl, the l, 2-methyl-4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl and 5-hexenyl,

"cycloalkyl group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl and substituted;

"sanirovano lower alkenylphenol group" such as 2-cyanobutane;

"Uralkaliy group, such as benzyl, alpha-naphthylmethyl, beta-naphthylmethyl, intermetal, phenantrolinate, anthranilates, diphenylmethyl, triphenylmethyl, 1-phenethyl, 2-phenethyl, 1-naphtalate, 2-naphtalate, 1-phenylpropyl, 2-phenylpropyl, 3-phenylpropyl, 1-naftilamin, 2-naftilamin, 3-naftilamin, 1-phenylbutyl, 2-phenylbutyl, 3-phenylbutyl, 4-phenylbutyl, 1-naphthylmethyl, 2-naphthylmethyl, 3-naphthylmethyl, 4-naphthylmethyl, 1-fenilpentil, 2-fenilpentil, 3-fenilpentil, 4-fenilpentil, 5-fenilpentil, 1-naphthylmethyl, 2-naphthylmethyl, 3-naphthylmethyl, 4-naphthylmethyl, 5-naphthylmethyl, 1-phenylhexa, 2-phenylhexa, 3-phenylhexa, 4-phenylhexa, 5-phenylhexa, 6-phenylhexa, 1-nattinger, 2-nattinger, 3-nattinger, 4-nattinger, 5-nattinger and 6-nattinger;

"Uralkaliy group in which the specified aryl ring substituted by a nitro-group or halogen atom", such as 4-Chlorobenzyl, 2-(4-nitrophenyl)ethyl ortho-nitrobenzyl, 4-nitrobenzyl, 2,4-dinitrobenzyl and 4 the ilen group, substituted lower alkyl group, a halogen atom or a nitro-group" such as 2-were, 2,6-dimetilfenil, 2-chlorophenyl, 4-chlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, 2-bromophenyl, 4-nitrophenyl and 4-chloro-2-nitrophenyl, preferably "lower alkyl group", a "lower alkyl group substituted by a cyano", "Uralkaliy group" or "Uralkaliy group in which the specified aryl ring substituted by a nitro-group or halogen atom, more preferably 2-cyanoethylene group, 2,2,2-trichlorethylene group or benzyl group.

"Alkoxygroup having from 1 to 4 carbon atoms", R3and R, or the alpha group in the above formula (1) or (2) may include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy or ethoxypropan.

The protective group of the protected mercaptopropyl" Alfa-group in the above formula (1) or (2) may include, in addition to the protective groups of hydroxyl, mentioned above, "a group that forms a disulfide", such as allylthiourea, for example methylthio, ethylthio, tert-butylthio and kalkiliya, such as benzylthio, preferably "aliphatic acyl group or aromatic acyl group", is up>4or the alpha group in the above formula (1) or (2) may include methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutyric, sec-butylthio and tert-butylthio, preferably methylthio or ethylthiourea.

The protective group of the protected amino group", the alpha group in the above formula (1) or (2) may include aliphatic acyl group, such as alkylcarboxylic group, such as formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, pivaloyl, valeryl, isovaleryl, octanoyl, nonanoyl decanoyl, 3-methylnonanoic, 8-methylnonanoic, 3-ethyloctanoic, 3,7-dimethyloctane, undecanoyl, dodecanoyl, tridecanol, deletion, pentadecanol, hexadecanol, 1-methylpentanol, 14-methylpentadiene, 13,13-dimethylcarbamoyl, heptadecanoyl, 15-methylhexadecanoic, octadecanoyl, 1-methylheptadecyl, nonadecanoic, eicosanoic and heneicosanol, carboxypropanoyl alkylcarboxylic group, such as succinoyl, glutaryl and adipoyl, halogen(lower)alkylcarboxylic group such as chloroacetyl, dichloroacetyl, trichloroacetyl and TRIFLUOROACETYL, lower alkoxy(lower)alkylcarboxylic group, such as methoxyacetyl, and unsaturated alkylcarboxylic group, such as (E)-2-methyl-2-butene the beta naphtol, haloidalkyls group, such as 2-bromobenzoyl and 4-chlorobenzoyl, lower alkilirovanny arylcarbamoyl group, for example 2,4,6-trimethylbenzoyl and 4-toluoyl, lower alkoxycarbonyl arylcarbamoyl group, for example 4-anisoyl, carboxypropanoyl arylcarbamoyl group, such as 2-carboxybenzoyl, 3-carboxybenzoyl and 4-carboxybenzoyl, nitrated arylcarbamoyl group, for example 4-nitrobenzoyl and 2-nitrobenzoyl, lower alkoxycarbonylmethyl arylcarbamoyl group, such as 2-(methoxycarbonyl)benzoyl and anilinophenol arylcarbamoyl group, for example 4-phenylbenzyl;

"lower alkoxycarbonyl group, such as methoxycarbonyl, etoxycarbonyl, tert-butoxycarbonyl and isobutoxide;

"lower alkoxycarbonyl group substituted by a halogen atom, or three(lower)alkylsilane group" such as 2,2,2-trichlorocyanuric and 2-trimethylsilylethynyl;

"altneratively group, such as vinyloxycarbonyl and allyloxycarbonyl, and

"aracelikarsaalyna group in which the specified aryl ring may be substituted by lower alkoxy or nitro-group", such as benzyloxycarbonyl, 4-methoxybenzenesulfonyl, 3,4-dimethoxybenzenesulfonamide allgroups", more preferably benzoylpropionic.

"Amino group, a substituted alkyl group having from 1 to 4 carbon atoms", R3and R4or the alpha group in the above formula (1) or (2) may include methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, sec-butylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamine, di(sec-butyl)amino and di(tert-butyl)amino, preferably methylamino, ethylamino, dimethylamino, diethylamino or diisopropylamino.

"Cyanoacetylurea having from 1 to 5 carbon atoms", R3and R4in the above formula (1) represents a group in which the above-described "alkoxygroup having from 1 to 4 carbon atoms", substituted by cyano, such group may include cyanoethoxy, 2-cyanoethoxy, 3 cyanopropionic, 4-cyanobutane, 3-cyano-2-methylpropoxy or 1-cyanomethyl-1,1-dimethylmethoxy, preferably 2-senatorship.

"An alkyl group having from 1 to 4 carbon atoms alpha group in the above formula (1) or (2) may include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl, preferably methyl or ethyl group is, the fact bromine or iodine atom, preferably fluorine atom or chlorine atom.

Preferred groups of the purine-9-ilen group" and "substituted purine-9-ilen group" In the above formula (1) or (2) may include, in General, 6-aminopurin-9-ilen (i.e. adenine), 6-aminopurin-9-ilen, the amino group of which is protected, 2,6-diaminopurine-9-ilen, 2-amino-6-globulin-9-ilen, 2-amino-6-globulin-9-ilen, the amino group of which is protected, 2-amino-6-toburen-9-ilen, 2-amino-6-toburen-9-ilen, the amino group of which is protected, 2-amino-6-bromopurine-9-ilen, 2-amino-6-bromopurine-9-ilen, the amino group of which is protected, 2-amino-6-hydroxypurine-9-ilen (i.e. guanine), 2-amino-6-hydroxypurine-9-ilen, the amino group of which is protected, 2-amino-6-hydroxypurine-9-ilen, in which the amino - and hydroxy-group-protected, 6-amino-2-methoxypurine-9-ilen, 6-amino-2-globulin-9-ilen, 6-amino-2-toburen-9-ilen, 2,6-dimethoxyfuran-9-ilen, 2,6-dichloropurine-9-ilen or 6-mercaptopurine-9-ilen, more preferably 6-benzylaminopurine-9-ilen, adenine, 2 isobutylamino-6-hydroxypurine-9-ilen or guanidino group.

Preferred groups 2-oxopyrimidine-1-ilen group" and "substituted 2-oxopyrimidine-1-ilen group" In the above formula (1) or gruppa of which is protected, 2-oxo-4-amino-5-ftorpirimidinu-1-ilen, 2-oxo-4-amino-5-ftorpirimidinu-1-ilen, the amino group of which is protected, 4-amino-2-oxo-5-chloropyrimidine-1-ilen, 2-oxo-4-methoxypiperidine-1-ilen, 2-oxo-4-mercaptopyrimidine-1-ilen, 2-oxo-4-hydroxypyrimidine-1-ilen (i.e. orcinol), 2-oxo-4-hydroxy-5-methylpyrimidin-1-ilen (i.e. teminal) or 4-amino-5-methyl-2-oxopyrimidine-1-ilen (i.e., 5-methylcytosine) group, more preferably 2-oxo-4-benzoylpiperidine-1-ilen, cytosine, timini, orcinol, 2-oxo-4-benzoylamine 5-methylpyrimidin-1-ilen, or 5-methylcytosine group.

The term "nucleoside analog" refers to an unnatural type "nuke" in which purine or pyrimidine group attached to the sugar.

The term "oligonucleotide analog" refers to an unnatural type "oligonucleotide" derivative, in which from 2 to 50 "nucleosides", which may be the same or different, are joined by devernay communication phosphoric acid, and such counterparts may preferably include sugar derivatives, in which the functional group modified sugar; tianye derivatives, in which diperna communication phosphoric acid tiatearaway; ester products in which the end of uhrinova basis is amidinophenoxy, more preferably sugar derivatives, in which the functional group modified sugar, and tianye derivatives, in which diperna functional group of phosphoric acid is titrovanie.

The term "their salts" refers to salts of the compounds (1) of the present invention because they can be converted into salts, and such salts may preferably include inorganic salts, for example metal salts, such as alkali metal salts, for example salts of sodium, potassium salts and lithium salts, salts of alkaline earth metals, for example calcium salts and magnesium salts, aluminum salts, iron salts, zinc salts, salts of copper, Nickel salts and cobalt salts; amine salts such as inorganic salts, for example ammonium salts, organic salts, for example, salts of tert-octylamine, salt dibenzylamine, salt of the research, glucosamine salt, salt of ester of alkylphenolic, salt, Ethylenediamine salt, N-methylglucamine salt of guanidine, salts diethylamine, salt, triethylamine salt dicyclohexylamine, salts of N,N'-dibenziletilendiaminom, salt chloroprocaine, salts of procaine, salt, diethanolamine, salts of N-benzylpenicillin, salt, piperazine salt of Tetramethylammonium and salt of Tris(hydroxymethyl)aminomethane, salt Neorganicheskie orodno acid, salts of Hydrobromic acid and salts idiscovered acids, salts of nitric acid, salt perchloro acids, salts of sulfuric acid, and salts of phosphoric acid; organic acid salts such as salts of lower alkanesulfonyl, for example salts methansulfonate, salt triftoratsetata and salt econsultancy, salt arylsulfonyl, for example salts benzosulfimide and salts of para-toluenesulfonic acid, salts of acetic acid, salts of malic acid, salts of fumaric acid, salts of succinic acid, salts of citric acid, salts of tartaric acid, salts of oxalic acid, and salts of maleic acid, and salts of amino acids such as salts of glycine, lysine salt, salt of arginine, ornithine salts, salts of glutamic acid and salts asparticularly acid.

Because the modified oligonucleotide or polynucleotide analogs of the present invention can be converted into salts, the term "their pharmacologically acceptable salts" refers to salts, and such salts may preferably include inorganic salts, for example metal salts, such as alkali metal salts, for example sodium salts, potassium salts, lithium salts, alkaline earth metals, for example calcium salts and magnesium salts, their salts, for example ammonium salts, organic salts, for example salts of tert-octylamine, salt dibenzylamine, salt of the research, glucosamine salt, salt of ester of alkylphenolic, salt, Ethylenediamine salt, N-methylglucamine salt of guanidine, salts diethylamine, salt, triethylamine salt dicyclohexylamine, salts of N,N'-dibenziletilendiaminom, salt chloroprocaine, salts of procaine, salt, diethanolamine, salts of N-benzylpenicillin, salt, piperazine salt of Tetramethylammonium, and salts of Tris(hydroxymethyl)aminomethane, inorganic salts, such as salts kaleidograph acids, for example salts of hydrofluoric acid, salt, hydrochloric acid salt, Hydrobromic acid salt idiscovered acids, salts of nitric acid, salt perchloro acids, salts of sulfuric acid, and salts of phosphoric acid; organic acid salts such as salts of lower alkanesulfonyl, for example salts methansulfonate, salt triftoratsetata and salt econsultancy, salt arylsulfonyl, for example salts benzosulfimide and salts of para-toluenesulfonic acid, salts of acetic acid, salts of malic acid, salts of fumaric acid, salts of succinic acid, salts of citric acid, salts of tartaric acid, salts of oxalic Kalotina, salt of glutamic acid and salts asparticularly acid.

Of the compounds (1) and their salts of the present invention preferred compounds may include:

(1) compounds in which R1represents a hydrogen atom, an aliphatic acyl group, aromatic acyl group, a methyl group substituted by from 1 to 3 aryl groups, a methyl group substituted by from 1 to 3 aryl groups, the aryl ring of which is substituted by lower alkyl, lower alkoxygroup, a halogen atom or a cyano, or a silyl group, and their salts;

(2) compounds in which R1represents a hydrogen atom, acetyl group, benzoyloxy group, benzyl group, para-methoxybenzyloxy group, dimethoxytrityl group, mono-methoxytrityl group or tert-butyldiphenylsilyl group, and their salts;

(3) compounds in which R2represents a hydrogen atom, an aliphatic acyl group, aromatic acyl group, a methyl group substituted by from 1 to 3 aryl groups, a methyl group substituted by from 1 to 3 aryl groups, the aryl ring of which is substituted by lower alkyl, lower alkoxy, halogen atom or cyano, silyl group, phosphamidon the

(4) compounds in which R2represents a hydrogen atom, acetyl group, benzoyloxy group, benzyl group, para-methoxybenzyloxy group, tert-butyldiphenylsilyl group, -P(OC2H4CN)(N(CH3)2), -P(och3)(NCH(CH3)2), fastonline group, or 2-chloraniline, or 4-chloraniline group of phosphoric acid, and their salts;

(5) compounds in which a represents a methylene group, and their salts;

(6) compounds in which a represents a 6-aminopurin-9-ilen (i.e. adenine), 6-aminopurin-9-ilen group, the amino group of which is protected, 2,6-diaminopurine-9-ilen, 2-amino-6-globulin-9-ilen, 2-amino-6-globulin-9-ilen group, the amino group of which is protected, 2-amino-6-toburen-9-ilen, 2-amino-6-toburen-9-ilen group, the amino group of which is protected, 2-amino-6-bromopurine-9-ilen, 2-amino-6-bromopurine-9-ilen, amino group which is protected, 2-amino-6-hydroxypurine-9-ilen (i.e. guanine), 2-amino-6-hydroxypurine-9-ilen, amino group which is protected, 2-amino-6-hydroxypurine-9-ilen, amino group and hydroxy-group of which is protected, 6-amino-2-methoxypurine-9-ilen, 6-amino-2-globulin-9-ilen, 6-amino-2-toburen-9-ilen, 2,6-dimethoxyfuran-9-ilen, 2,6-dichloropurine, the amino group of which is protected, 2-oxo-4-amino-5-ftorpirimidinu-1-ilen, 2-oxo-4-amino-5-ftorpirimidinu-1-ilen, the amino group of which is protected, 4-amino-2-oxo-5-chloropyrimidine-1-ilen, 2-oxo-4-methoxypiperidine-1-ilen, 2-oxo-4-mercaptopyrimidine-1-ilen, 2-oxo-4-hydroxypyrimidine-1-ilen (i.e. orcinol), 2-oxo-4-hydroxy-5-methylpyrimidin-1-ilen (i.e. teminal), 4-amino-5-methyl-2-oxopyrimidine-1-ilen (i.e., 5-methylcytosine) group, or 4-amino-5-methyl-2-oxopyrimidine-1-ilen group in which the amino group is protected, and their salts, and

(7) compounds in which a represents a 6-benzylaminopurine-9-ilen, adenine, 2-isobutylamino-6-hydroxypurine-9-ilen, guanidino, 2-oxo-4-benzoylpiperidine-1-ilen, casinillo, 2-oxo-5-methyl-4-benzoylpiperidine-1-ilen, 5-methylcytosine, brazenly or ciminillo group, and their salts.

In the above embodiments (1) and (2), (3) and (4) or (6) and (7) are preferred compounds in increasing numbers, and in the formula (1) are preferred compounds obtained by the optional choice of R1from (1) and (2), optional choice of R2from (3) and (4), optional choice And from (5) and the optional choice of (6) and (7) or by optional the E. of the following groups.

A group of compounds:

2'-O,4'-C-teleguinin,

2'-O,4'-C-utilizationin,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-6-N-benzyladenine,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-2-N-isobutylamine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamine

2'-O,4'-C-ethylene-2-N-isobutylamine,

2'-O,4'-C-ethylene-6-N-benzyladenine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine-3'-O-(2-Tianeti N,N-aminobutiramida)phosphoamide,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamino-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide,

2'-O,4'-C-ethyleneimine,

2'-O,4'-C-ethylene-5-methyluridine,

2'-O,4'-C-atransitive,

2'-O,4'-C-ethylene-5-methylcytidine,

3',5'-di-O-benzyl-2'-O,4'-C-ethyleneimine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethyleneimine,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-5-methyluridine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-5-methyluridine,

3',5' -di-O-benzyl-2'-O,4'-C-ethylene-4-N-benzoylation,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation,

3', 5'-di-O-benzyl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine,

2'-O,4'-C-ethylene-4-N-benzoylation,

2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine,

5,4'-C-ethylene-5-methyluridine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide and

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide.

Of oligonucleotide analogues containing one, or two or more structures of the formula (2), and their pharmacologically acceptable salts of the present invention preferred compounds may include:

(8) oligonucleotide analogues in which a represents a methylene group, and their pharmacologically acceptable salts;

(9) oligonucleotide analogues, which represents a 6-aminopurin-9-ilen (i.e. adenine), 6-aminopurin-9-ilen, amino group which is protected, 2,6-diaminopurine-9-ilen, 2-amino-6-globulin-9-ilen, 2-amino-6-globulin-9-ilen, amino group which is protected, 2-amino-6-toburen-9-ilen, 2-amino-6-toburen-9-ilen, amino group which is protected, 2-amino-6-bromopurine-9-ilen, 2-amino-6-bromopurine-9-ilen, the amino group of which is protected, 2-amino-6-hydroxypurine-9-ilen (i.e. guanine), 2-amino-6-hydroxypurine-9-ilen, the amino group of which is protected, 2-amino-6-hydroxypurine-9-ilen, amino group and hydroxyl group which is protected, 6-amino-2-methoxypurine-9-ilen, 6-amino-2-chlorphenol, 2-oxo-4-aminopyrimidine-1-ilen (i.e. cytosine), 2-oxo-4-aminopyrimidine-1-ilen, the amino group of which is protected, 2-oxo-4-amino-5-ftorpirimidinu-1-ilen, 2-oxo-4-amino-5-ftorpirimidinu-1-ilen, the amino group of which is protected, 4-amino-2-oxo-5-chloropyrimidine-1-ilen, 2-oxo-4-methoxypiperidine-1-ilen, 2-oxo-4-mercaptopyrimidine-1-ilen, 2-oxo-4-hydroxypyrimidine-1-ilen (i.e. arallel), 2-oxo-4-hydroxy-5-methylpyrimidin-1-ilen (i.e. teminal), 4-amino-5-methyl-2-oxopyrimidine-1-ilen (i.e., 5-methylcytosine) group, or 4-amino-5-methyl-2-oxopyrimidine-1-ilen group, the amino group of which is protected, and their pharmacologically acceptable salts

and (10) oligonucleotide analogues, which represents a 6-benzylaminopurine-9-ilen, adreniline, 2-isobutylamino-6-hydroxypurine-9-ilen, guanidino, 2-oxo-4-benzoylpiperidine-1-ilen, casinillo, 2-oxo-5-methyl-4-benzoylpiperidine-1-ilen, 5-methylcytosine, brazenly or ciminillo group, and their pharmacologically acceptable salts.

In the above embodiments (9) and (10) are more preferred oligonucleotide analogues, with increasing numbers, and in the formula (2) are preferred are oligonucleotide analogs obtained by neobazine and their pharmacologically acceptable salts.

Specific compounds included in the number of connections by the above formula (1) of the present invention are listed in Tables 1 and 2. However, compounds of the present invention is not limited to these compounds.

In Table 1 and Table 2 No. Conn. represents a connection number given as example. Me represents a methyl group, PG is a benzyl group, Bz represents benzoyloxy group, RPS represents the p-methoxybenzyloxy group, Tg represents triphenylmethyl group, MTG is a 4-methoxycarbonylmethylene (monoethoxylate) group, DMTr represents 4,4'-dimethoxytrityl (dimethoxytrityl) group, Tmtg represents 4,4',4"-trimethoxytrityl (trimethoxytrityl) group, TMS represents trimethylsilyl group, TBDMS represents a tert-butyldimethylsilyloxy group, TBDPS represents a tert-butyldiphenylsilyl group and TIPS is triisopropylsilyl group.

In the above tables 1 and 2, the preferred compounds are compounds(1-5), (1-7), (1-23), (1-24), (1-31), (1-35), (1-39), (1-43), (1-49), (1-51), (1-67), (1-68), (1-75), (1-79), (1-83), (1-87), (1-93), (1-95), (1-111), (1-112), (1-119), (1-12, 2-15), (2-19), (2-22), (2-27), (2-31), (2-34), (2-39), (2-43), (2-46), (2-51), (2-55), (2-57), (2-58), (2-59), (2-60), (2-66), (2-71), (2-75), (2-78), (2-83), (2-87), (2-90), (2-95), (2-99), (2-102), (2-107), (2-111), (2-113), (2-114), (2-115), (2-116), (2-122), (2-127), (2-131), (2-134), (2-139), (2-143), (2-146), (2-151), (2-155), (2-158), (2-163), (2-167), (2-169), (2-170), (2-171), (2-172), (2-178), (2-183), (2-187), (2-190), (2-195), (2-199), (2-202), (2-207), (2-211), (2-214), (2-219), (2-223), (2-225), (2-226), (2-233), (2-234), (2-235) or (2-236),

more preferable compounds may include:

2'-O,4'-C-teleguinin (1-5),

2'-O,4'-C-utilizationin (1-7),

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-6-N-benzyladenine (1-23),

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-2-N-isobutylamine

(1-24),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine

(1-31),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamine (1-35),

2'-O,4'-C-ethylene-2-N-isobutylamine (1-177),

2'-O,4'-C-ethylene-6-N-benzyladenine (1-178),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamino-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoramidite (1-185),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (1-186),

2'-O,4'-C-ethylenedi (2-1),

2'-O,4'-C-ethylene-5-methyluridine (2-2),

2'-O,4'-C-atransition (2-3),

2'-O,4'-C-ethylene-5-methylcytidine (2-4),

3',5'-di-O-benzyl-2'-O,4'-C-ethylenedi (2-10),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylenedi (2-15),

'-di-O-benzyl-2'-O,4'-C-ethylene-4-N-benzoylation (2-34),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation (2-39),

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine (2-46),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine (2-51),

2'-O,4'-C-ethylene-4-N-benzoylation (2-225),

2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine (2-226),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylenedi-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (2-233),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-5-methyluridine-3'-O-(2-Tianeti-N, N-aminobutiramida)phosphoamide (2-234),

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (2-235)

and 5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine-3'-O-(2-Tianeti-N, N-aminobutiramida)phosphoamide (2-236).

Connection (1) of the present invention can be obtained in accordance with the method As described below.

In the method And X represents a protective group; Y represents a protective group; a has the same meaning defined above, while In1represents a purine-9-ilen group, a substituted purine-9-ilen group or substituted 2-oxopyrimidine-1-ilen group, and these substituents selected from the above R3and R4, takaeclipse a purine-9-ilen group, substituted purine-9-ilen group or substituted 2-oxopyrimidine-1-ilen group, and these substituents selected from the above R3and R4however , with the exception of unprotected amino group "amino group which may be protected; R7represents a group which forms tsepliaeva group, and R8represents an aliphatic acyl group having from 1 to 4 carbon atoms.

The protective group X is the same group as the protective group of hydroxyl in the above R1.

The protective group for Y is the same group as the protective group of hydroxyl in the above R2.

"The group that forms tsepliaeva group, R7may include lower alkylsulfonyl group, such as methanesulfonyl and econsultancy; halogen-substituted lower alkylsulfonyl group, such as trifloromethyl, and arylsulfonyl group such as para-toluensulfonyl, preferably methanesulfonyl group or p-toluensulfonyl group.

"Aliphatic acyl group containing from 2 to 4 carbon atoms, R8may include acetyl, propionyl, butyryloxy group and the like, predpochtitelney stage is intended to obtain compound (4) by reacting compound (3), which can be obtained by methods b to D described below, with a reagent for introducing tsepliaeva group in the presence of a basic catalyst in an inert solvent.

Used for this purpose solvents may include aliphatic hydrocarbons such as hexane, heptane, ligroin and petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl, butyl acetate and diethylmalonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and dimethyl ether of diethylene glycol; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone and cyclohexanone; nitro compounds such as nitroethane and nitrobenzene; NITRILES such as acetonitrile and isobutyronitrile; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone and hexamethylphosphorotriamide; sulfoxidov, such as sulfolan, and derivatives of pyridine, preferably pyridine.

The main catalyst male introduction tsepliaeva group may include alkylsulfonate, such as methanesulfonate and acanaloniidae, and arylsulfonate, such as p-toluensulfonate, preferably methanesulfonate and p-toluensulfonate.

The reaction temperature varies depending on the starting material, solvent, reagent for the introduction of tsepliaeva group and a basic catalyst, but it usually ranges from 0 to 50C, preferably from 10 to 40C.

The time of interaction varies depending on the starting material, solvent, reagent for the introduction of tsepliaeva group, the basic catalyst and the reaction temperature, but it usually takes from 10 minutes to 24 hours, preferably from 1 to 10 hours

After the interaction, the desired compound (4) this reaction receive, for example, by neutralization of the reaction solution, the concentration of the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary(stage-2)

This stage is intended to obtain compound (5) by reacting compound (4) obtained in stage a-1, with an acid anhydride, in the presence of an acid catalyst in a solvent.

Used for this purpose solvents may include ethers such as diethyl ether, tetrahydrofuran and dioxane; NITRILES, such as acetonitrile and isobutyronitrile; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone and hexamethylphosphorotriamide, and organic acids such as acetic acid, preferably acetic acid.

The acid catalysts may include inorganic acids such as hydrochloric acid, sulfuric acid and nitric acid, preferably sulfuric acid (especially sulfuric acid).

The anhydrides of acids may include the anhydride of a lower aliphatic carboxylic acid, such as acetic anhydride and propionic anhydride, preferably acetic anhydride.

The reaction temperature varies depending on the starting material, solvent, acid catalyst and the acid anhydride and usually it ranges from 0 to the applied depending on the source material, solvent, acid catalyst, the acid anhydride and the reaction temperature, but it usually ranges from 10 min to 12 h, preferably from 30 minutes to 3 hours

After the interaction, the desired compound (5) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatography on a column of silica gel, etc.,

(Stage-3)

This stage is intended to obtain compound (6) by reacting compound (5) obtained in stage a-2, with trimethylsilylethynyl connection corresponding to the purine or pyrimidine, which can have the desired Deputy received in accordance with reference (N. Vorbrggen, K. Krolikiewicz and Century Bennua, Chem. Weg., so 114, S. 1234-1255 (1981)), in the presence of acid catalyst in an inert solvent.

Used for this purpose solvents can switch risty methylene, chloroform, carbon tetrachloride, 1,2-dichloroethane, chlorobenzene and dichlorobenzene; NITRILES, such as acetonitrile and isobutyronitrile; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone and hexamethylphosphorotriamide; carbon disulfide; preferably 1,2-dichloroethane.

The acid catalysts may include catalysts are Lewis acid, such as ll3, SnCl4, TiCl4, ZnCl2BF3, trimethylsilyltrifluoromethane, preferably trimethylsilyltrifluoromethane.

The reaction temperature varies depending on the starting material, solvent and acid catalyst, but it usually ranges from 0 to 100C, preferably from 50 to 80C.

The time of interaction varies depending on the starting material, solvent, acid catalyst and reaction temperature, but it usually takes from 1 to 24 h, preferably from 1 to 8 hours

After the interaction, the desired compound (6) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent not miscible with water, such as Etisalat magnesium and removal of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatography on a column of silica gel, etc.,

(Stage-4)

This stage is intended to obtain compound (1A) of the present invention by cyclization of compound (6) obtained in stage a-3, in the presence of a basic catalyst in an inert solvent.

Used for this purpose solvents include water; pyridine derivatives; acetonitrile, such as acetonitrile and isobutyronitrile; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone and hexamethylphosphorotriamide, and mixtures thereof, preferably a mixture of water and pyridine.

Applied here, the basic catalyst may include hydroxides of alkali metals such as sodium hydroxide and potassium hydroxide; carbonates of alkali metals such as sodium carbonate and potassium carbonate; alkoxides of alkali metals such as sodium methoxide and ethoxide sodium; aqueous ammonia, preferably hydroxides of alkali metal (especially sodium hydroxide).

The reaction temperature varies depending on the source>the, preferably from 10 to 30C.

The time of interaction varies depending on the starting material, solvent, basic catalyst and the reaction temperature, but usually it ranges from 1 min to 5 h, preferably from 1 to 30 minutes.

After the interaction, the desired compound (1A) this reaction receive, for example, by neutralization of the reaction solution, the concentration of the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

(Stage-5)

This stage is intended to obtain compound (1b) by reacting the compound (1A) obtained in stage a-4, removing the protecting reagent, in an inert solvent.

Method of removing protection vary depending on the type of the protective group and is not specifically limited if it does not bring the ic Synthesis" (Theodora W. Greene and Peter G. M. Wuts, 1999, published in A Wiley-Interscience Publication).

Specifically, the removal method of protection can be performed by the following methods, in the case where the protective group is a (1) "aliphatic acyl group or aromatic acyl group", (2) "methyl group substituted by from 1 to 3 aryl groups" or "methyl group substituted by from 1 to 3 aryl groups, the aryl ring of which is substituted by lower alkyl, lower alkoxy, halogen atom or panography", or (3) "silyl group".

(1) if, when the protective group is an aliphatic acyl group or aromatic acyl group, the reaction of the removal of protection is usually carried out by treating the compound base in an inert solvent.

Used for this purpose, the solvent is not specifically limited, if they are easily mixed with water, does not inhibit the reaction and to some extent dissolve the starting material, and may include aqueous or anhydrous amides, such as dimethylformamide and dimethylacetamide; halogenated hydrocarbons such as methylene chloride, chloroform, 1,2-dichloroethane or carbon tetrachloride; ethers, such as tetrahydrofuran, diethyl ether and dioxane; precht hydroxides of alkali metals, such as lithium hydroxide, sodium hydroxide and potassium hydroxide; carbonates of alkali metals such as sodium carbonate and potassium carbonate; alkoxides of alkali metals such as sodium methoxide and ethoxide sodium, and ammonia solution, such as aqueous ammonia and ammonia in a solution of methanol.

The reaction temperature is 0 to 60C, preferably from 20 to 40C.

Interaction time is from 10 minutes to 24 hours, preferably from 1 to 3 hours.

After the interaction, the desired compound (1b) this reaction receive, for example, by neutralization of the reaction solution, the concentration of the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example, recrystallization and chromatography on a column of silica gel, etc.,

(2) In the case when the protective group is a "methyl group substituted by one to three and the x substituted lower alkyl, lower alkoxy-, halogen atom or cyano, the reaction is usually carried out in an inert solvent using a reducing agent.

Used for this purpose solvents can preferably include alcohols, such as methanol, ethanol and isopropanol; ethers, such as diethyl ether, tetrahydrofuran and dioxane; aromatic hydrocarbons, such as toluene, benzene and xylene; aliphatic hydrocarbons such as hexane and cyclohexane; esters such as ethyl acetate and propyl; organic acids such as acetic acid, or a mixture of these organic solvents and water.

Used for this purpose rejuvenating agents are not specifically limited, if they are usually used for catalytic reduction, and they may include preferably palladium on charcoal, Raney Nickel, platinum oxide, platinum black, rhodium-aluminum oxide, triphenylmethylchloride and palladium on barium sulphate.

Pressure is not specially limited, but usually it ranges from 0.1 to 1 MPa (1-10 ATM).

The reaction temperature is 0 to 60C, preferably from 20With up to 40C.

Time shall tell (1b) this reaction get for example, by removing the reducing agent from the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent. Thus obtained target product can be further purified, if necessary, in the traditional way, for example, recrystallization, chromatography on a column of silica gel, etc.,

When the protective group is a "methyl group substituted by three aryl groups", i.e. trailing group, the reaction of the removal of protection can also be performed with the use of acids.

In this case, used here solvents may include aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, 1,2-dichloroethane, chlorobenzene and dichlorobenzene; NITRILES, such as acetonitrile and isobutyronitrile; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone and hexamethylphosphorotriamide, and organic acids, Tokyo tert-butanol).

The acid may preferably include acetic acid or triperoxonane.

The reaction temperature is 0 to 60C, preferably from 20 to 40C.

Interaction time is from 10 minutes to 24 hours, preferably from 1 to 3 o'clock

After the interaction, the desired compound (1b) this reaction receive, for example, by neutralizing the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization, chromatography on a column of silica gel, etc.,

(3) if, when the protective group is a "silyl group", then usually it can be removed by treating the compound that forms the anion of fluorine, such as tetrabutylammonium, hydrofluoric acid, hydrofluoric acid-pyridine and potassium fluoride, or organic acids such as acetic acid, IU the acid, or inorganic acid such as hydrochloric acid.

When the protective group is removed under the action of the anion of fluorine, the reaction sometimes activate by adding to the mixture of organic acids, such as formic acid, acetic acid and propionic acid.

Used for this purpose, the solvent is not specifically limited if they do not slow the reaction and to some extent dissolve the starting material, and may preferably include ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and dimethyl ether of diethylene glycol; NITRILES, such as acetonitrile and isobutyronitrile; water; organic acids such as acetic acid, and mixtures thereof.

The reaction temperature is from 0 to 100C, preferably from 20 to 70C.

Interaction time is from 5 min to 48 h, preferably from 1 to 24 hours.

After the interaction, the desired compound (1b) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer, is thus the target product can be further purified, if necessary, the traditional way, for example by recrystallization, chromatography on a column of silica gel, etc.,

Stage A-6

This stage is intended to obtain compound (1C) of the present invention by reacting compound (1b) obtained in stage A-5, to remove the protecting reagent, in an inert solvent.

Method of removing protection vary depending on the type of the protective group and is not specifically limited if it does not result in other adverse reactions, and may be implemented, for example, by the method described in the book "Protective Groups in Organic Synthesis" (Theodora W. Greene and Peter G. M. Wuts, 1999, published in A Wiley-Interscience Publication).

In particular, the removal method of protection can be implemented in the following way, in the case where the protective group is an aliphatic acyl group or aromatic acyl group.

Specifically, the removal method of protection is usually carried out by reacting with a base in an inert solvent, in the case where the protective group is an aliphatic acyl group or aromatic acyl group.

Used for this purpose, the solvent is not specifically limited, if they are easily mixed with evadnie alcohols, such as methanol and ethanol; amides such as dimethylformamide and dimethylacetamide; halogenated hydrocarbons such as methylene chloride, chloroform, 1,2-dichloroethane or carbon tetrachloride, ethers, such as tetrahydrofuran, diethyl ether and dioxane; preferred alcohols, preferably methanol.

Applied here, the base can include hydroxides of alkali metals such as lithium hydroxide, sodium hydroxide and potassium hydroxide; carbonates of alkali metals such as sodium carbonate and potassium carbonate; alkoxides of alkali metals such as sodium methoxide and ethoxide sodium, and ammonia, preferably ammonia.

The reaction temperature is 0 to 50C, preferably from 10 to 40C.

Interaction time is from 10 minutes to 24 hours, preferably from 10 minutes to 15 hours. After the interaction, the desired compound (1C) get this reaction, for example by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation rasti, traditional means, such as recrystallization and chromatographic column with silica gel, etc.,

The above intermediate compound (3) can be obtained by methods b through D below.

In methods B - D X and Y have the same meanings defined above; R9represents a group which forms tsepliaeva group; E represents ethylene, trimethylene or tetramethylene group, and Z is a simple bond, methylene or ethylene group.

The group, which forms tsepliaeva group, R9may include group described above for R7preferably trifloromethyl group.

R11and R12are the same and represent a hydrogen atom or taken together form an oxygen atom.

In the case when R11and R12taken together form an oxygen atom, R10represents an alkyl group having from 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl, preferably methyl group. In the case when R11and R12are the same and represent a hydrogen atom, RCimetidina group; benzoyloxymethyl group, such as benzoyloxymethyl group, uraleleketro group, such as benzoyloxymethyl group; alkoxylalkyl group, such as methoxyethoxymethyl group; a silyl group, such as trimethylsilyl, tert-butyldimethylsilyl, diphenylmethylsilane, diphenylmethylsilane, diphenylmethylsilane and phenyldimethylsilane.

Compound (7), i.e., the source material used in method b or method C, can be obtained as follows.

Specifically, the connection corresponding to the connection (6), in which functional group X is a hydrogen atom, is obtained from 1,1,5,6-diisopropylidene-D-glucose (on sale) in accordance with the described in the literature (R. D. Youssefyeh, J. P. H. Verheyden, J. G. Moffatt. J. Org. Chem., 44, 1301-1309 (1979)) and then the compound (6) can be obtained as described in the literature (I. Waga, I. Nishizaki, I. Miyakawa, H. Ohrui, H. Meguro, Biosci. Biotechnol. Biochem., 57, 1433-1438 (1993)) (in the case when X=benzyl).

Method In

Stage-1

This stage is intended to obtain compound (8) by reacting the compound (7) obtained above, with a reagent for introducing tsepliaeva group in the presence of basic is such as dimethylformamide and dimethylacetamide; halogenated hydrocarbons, such as methylene chloride, chloroform, 1,2-dichloroethane or carbon tetrachloride, ethers, such as tetrahydrofuran, diethyl ether and dioxane; preferred methylene chloride.

The basic catalyst may preferably include bases, such as triethylamine, pyridine and dimethylaminopyridine.

The reagent used to introduce tsepliaeva group may include preferably a chloride triftormetilfullerenov acid or anhydride triftormetilfullerenov acid.

The reaction temperature varies depending on the starting material, solvent and acid catalyst, but it is usually from -100 to -50C, preferably from -100 to -70C.

The time of interaction varies depending on the starting material, solvent, acid catalyst and reaction temperature, but it usually takes from 30 min to 12 h, preferably from 30 minutes to 3 hours

After the interaction, the desired compound (8) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, prom is ing and removal of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatography on a column of silica gel.

Stage-2

This stage is intended to obtain compound (9) by reacting compound (8) obtained in stage-1, with tianyoude reagent, in an inert solvent.

Used for this purpose solvents may include amides such as dimethylformamide and dimethylacetamide; halogenated hydrocarbons such as methylene chloride, chloroform, 1,2-dichloroethane or carbon tetrachloride; ethers, such as tetrahydrofuran, diethyl ether and dioxane; acetonitrile; dimethyl sulfoxide and so on; the preferred amides (dimethylformamide).

Used here tianyoude reagent may include KCN, NaCN and trimethylsilane, preferably NaCN.

The reaction temperature varies depending on the starting material, solvent and tianyoude reagent, but it usually ranges from 0 to 100C, preferably from 30 to 70C.

The time of interaction varies depending on the starting material, solvent, cinerous/p>After the interaction, the desired compound (9) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

Stage-3

This stage is intended to obtain compound (10) by reacting compound (9) obtained in stage-2, with a regenerating agent in an inert solvent.

Used for this purpose solvents may include halogenated hydrocarbons such as methylene chloride, chloroform, 1,2-dichloroethane or carbon tetrachloride; aliphatic hydrocarbons such as hexane, heptane, ligroin and petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and Dimitrova the non; preferred halogenated hydrocarbons (particularly methylene chloride).

Used here, the reducing agent may include diisobutylaluminium and trichotillomania, preferably diisobutylaluminium.

The reaction temperature varies depending on the starting material, solvent and reducing agent, but it usually ranges from -100 to -50With, preferably -90 to -70C.

The time of interaction varies depending on the starting material, solvent, reducing agent and the reaction temperature, but it usually takes from 30 min to 12 h, preferably from 1 to 5 o'clock

After the interaction, the desired compound (10) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization to obtain compound (), one of the starting materials of the way And, by reacting compound (10) obtained in stage-3, with a regenerating agent in an inert solvent.

Used here solvents may include alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, Isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol and methylcellosolve, and acetic acid, the preferred alcohols (especially ethanol).

Used here, the reducing agent may include borohydride alkali metals, such as borohydride sodium and borohydride lithium; aluminiumhydride compounds such as sociallyengaged and laetrilelamygdalin, and boron, predpochtitelno borohydride sodium.

The reaction temperature varies depending on the starting material, solvent and reducing agent, but it usually ranges from 0 to 50C, preferably from 10 to 40C.

The time of interaction varies depending on the starting material, solvent, reducing agent and the reaction temperature, but it usually ranges from 10 min to 12 h, preferably from 30 minutes to 5 hours

After vzaimodeystvia, concentration of the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

The way To

Stage s-1

This stage is intended to obtain compound (11) by reacting compound (7) obtained in the above way, with an oxidizing agent in an inert solvent.

Used here solvents may include aliphatic hydrocarbons such as hexane, heptane, ligroin and petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl, butyl acetate and diethylmalonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran isobutylketone, isophorone and cyclohexanone; preferred halogenated hydrocarbons (particularly methylene chloride).

The oxidizing agent used herein may include a reagent Swarna (Swern) for oxidation reagent Dess-Martin oxidation, a complex of chromium trioxide, such as a complex of pyridine hydrochloride/chromium trioxide (chlorproma pyridinium and pyridinium dichromate), preferably the reagent Swarna for oxidation (namely, dimethylsulfoxide-oxalyl chloride).

The reaction temperature varies depending on the starting material, solvent and oxidizing agent, but it usually ranges from -100 to -50C, preferably from -100 to -70C.

The time of interaction varies depending on the starting material, solvent, oxidizing agent and the reaction temperature, but it usually takes from 30 min to 12 h, preferably from 1 to 5 o'clock

After the interaction, the desired compound (11) this reaction receive, for example, by decomposition of the oxidizing agent, the concentration of the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, seat to be further purified, if necessary, the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

Stage-2

This stage is intended to obtain compound (12) by reacting compound (11) obtained in stage s-1, with a reagent that increases the carbon chain in an inert solvent.

Used here solvents may include aliphatic hydrocarbons such as hexane, heptane, ligroin and petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl, butyl acetate and diethylmalonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and dimethyl ether of diethylene glycol, ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone and cyclohexanone; preferred halogenated hydrocarbons (particularly methylene chloride).

Used herein, the reagents can include the Wittig reagent (Wittig), reagent Horner-Emmons, reagent Peterson (Peterson), the system reactive Tebbe.

The reaction temperature varies depending on the starting material, solvent and reagent, increasing the carbon chain, but it usually ranges from -20 to 20With, preferably 0C.

The time of interaction varies depending on the starting material, solvent, reagent, increasing the carbon chain, and the reaction temperature, but it usually takes from 30 min to 12 h, preferably from 1 to 5 o'clock

After the interaction, the desired compound (12) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

Stage-3

This stage is intended to obtain compound (3A) by the selective introduction of a hydroxyl group at the terminal carbon atom by a double may include aliphatic hydrocarbons, such as hexane, heptane, ligroin and petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl, butyl acetate and diethylmalonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and dimethyl ether of diethylene glycol, ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone and cyclohexanone; preferred ethers (particularly tetrahydrofuran).

The reagent used in this reaction may include borane, vitaminbody, tert-vexillary, 9-BBN (9-borabicyclo[3.3.1]nonan), preferably 9-BBN.

The reaction temperature varies depending on the starting material, solvent and reagent, but it usually ranges from 0 to 50C, preferably from 10 to 40C.

The time of interaction varies depending on the starting material, solvent, reagent and reaction temperature, but it usually takes from 6 to 48 h, preferably 12 to 24 hours.

After vzaimodeistvie, add organic solvent not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

Method D

Stage D-1

This stage is intended to obtain compound (13) by reacting compound (11) obtained in stage s-1, with a reagent that increases the carbon chain in an inert solvent.

Used here solvents may include aliphatic hydrocarbons such as hexane, heptane, ligroin and petroleum ether; aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl, butyl acetate and diethylmalonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and dim lorexane; preferably ethers (particularly tetrahydrofuran), preferably halogenated hydrocarbons (particularly methylene chloride).

Used herein, the reagents can include the Wittig reagent and the reagent Horner-Einmons.

The reaction temperature varies depending on the starting material, solvent and reagent but it usually ranges from -20 to 40C, preferably from 0 to 20C.

The time of interaction varies depending on the starting material, solvent, reagent and reaction temperature, but it usually takes from 30 min to 12 h, preferably from 1 to 5 o'clock

After the interaction, the desired compound (13) this reaction receive, for example, by concentrating the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

This stage can be carried out in accordance with option (2) stage A-5. In the case when R10represents an optionally substituted benzyl group, and R11and R12are hydrogen atoms, the compound (3b) can be obtained directly at this stage.

Stage D-3

This stage is intended to obtain compound (3b), one of the starting materials of the way And, by reacting compound (14) obtained in stage D-2, with a regenerating agent.

(a) In the case when R11and R12taken together form an oxygen atom.

Used here solvents may include alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, Isobutanol, tert-butanol, isoamyl alcohol, diethylene glycol, glycerin, octanol, cyclohexanol and methylcellosolve, and acetic acid; the preferred alcohols (especially ethanol).

Used here, the reducing agent may include borohydride alkali metals such as lithium borohydride; aluminiumhydride compounds such as sociallyengaged and laetrilelamygdalin, and boron; preposterouswords and a reducing agent, but it usually ranges from 0 to 50C, preferably from 10 to 40C.

The time of interaction varies depending on the starting material, solvent, reducing agent and the reaction temperature, but it usually ranges from 10 min to 12 h, preferably from 30 minutes to 5 hours

After the interaction, the desired compound (3b) get this reaction, for example by decomposition of the reducing agent, concentration of the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization and chromatographic column with silica gel, etc.,

(b) In the case when R11and R12represent hydrogen atoms, and R10is a group that is different from benzyl group.

In the case when R10is a silyl group, this stage can be carried out in accordance with the method (3 is Kilroy group, such as methoxymethyl group; arylcarboxylic group, such as benzoyloxymethyl group, or aralkylamines group, such as benzoyloxymethyl group, and alkoxylalkyl group, such as methoxyethoxymethyl group used an acid catalyst, and is used in this case, the acid catalyst may include an organic acid such as p-toluensulfonate acid, triperoxonane acid and dichloracetic acid and a Lewis acid such as F3and ll3.

Used here solvents may include aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, 1,2-dichloroethane, chlorobenzene and dichlorobenzene; NITRILES, such as acetonitrile and isobutyronitrile; amides, such as formamide, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone, N-methylpyrrolidinone and hexamethylphosphorotriamide, and carbon disulfide.

The reaction temperature varies depending on the starting material, solvent and acid catalyst, but it usually ranges from 0 to 50C, preferably from 10 to 40

After the interaction, the desired compound (3b) this reaction receive, for example, by neutralizing the reaction mixture, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example, by recrystallization, chromatographic column with silica gel, etc.,

Oligonucleotides containing modified nucleoside or its toutou derivative, can be obtained according to method E, which is described below, using the compounds (1) of the present invention.

In method E And has the same meaning as above; R13represents a protective group of hydroxyl (in particular, trityloxy group which may be substituted by a methoxy group); R13represents postonline group or a group formed by the interaction of monosubstituted chlorine(alkoxy)prednaznachaetsya to obtain compound (15) by reacting compound (1C), obtained in method A, with a protective reagent in an inert solvent.

Used here solvents preferably may include aromatic hydrocarbons such as benzene, toluene and xylene; halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene; esters such as ethyl formate, ethyl acetate, propyl, butyl acetate and diethylmalonate; ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxyethane and dimethyl ether of diethylene glycol; ketones, such as acetone, methyl ethyl ketone, methyl isobutyl ketone, isophorone and cyclohexanone; nitro compounds such as nitroethane and nitrobenzene; NITRILES, such as acetonitrile and isobutyronitrile; amides, such as formamide, dimethylformamide (DMF), dimethylacetamide and hexamethylphosphorotriamide; sulfoxidov, such as dimethyl sulfoxide and sulfolane; aliphatic tertiary amines such as trimethylamine, triethylamine and N-methylmorpholine, and aromatic amines such as pyridine and picoline; more preferred halogenated hydrocarbons (particularly methylene chloride) and aromatic amines, especially pyridine).

Used here is et to be removed under conditions of acidic or neutral environment, however, preferably, it may include triarylmethane, such as Fritillaria, monomethoxypolyethylene and dimethoxytrityl.

When as a protective reagent used triarylmethane, usually apply Foundation.

In this case, apply the base can include heterocyclic amines such as pyridine, dimethylaminopyridine and pyrrolidinone, and aliphatic tertiary amines, such as trimethylamine and triethylamine; the preferred pyridine, dimethylaminopyridine and pyrrolidinones.

When the solvent is used the liquid phase, because the very Foundation plays the role of an agent that binds acid, there is no need to add another base.

The reaction temperature varies depending on the starting material, solvent and reagent, but it is usually from 0 to 150C, preferably from 20 to 100C.

The time of interaction varies depending on the starting material, solvent and reaction temperature, but usually it ranges from 1 to 100 hours, preferably from 2 to 24 hours.

After the interaction, the desired compound (15) this reaction get nab the water, such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent.

Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization, chromatographic column with silica gel, etc.,

Stage E-2

This stage is intended to obtain compound (16) by reacting compound (15) obtained in Stage E-1, monosubstituted with chlorine(alkoxy)phosphines or disubstituted by alkoxysilane that are typically used for amidation in an inert solvent.

Used for this purpose, the solvent is not specifically limited if they do not slow the reaction and preferably may include ethers such as tetrahydrofuran, diethyl ether and dioxane, and halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, dichloroethane, chlorobenzene and dichlorobenzene.

Used here monosubstituted chlorine(alkoxy)phosphines may include phosphine derivatives, such as chlorine(morpholino)metaxiotis, chlorine(morpholino)canadaceftin, chlorine(dimethylamylamine)canadaceftin, preferably chlorine(morpholino)metaxiotis, chlorine(morpholino)canadaceftin, chlorine(diisopropylamino)metaxiotis and chlorine(diisopropylamino)canadaceftin.

When used monosubstituted chlorine(alkoxy)phosphines used acid binding agent, and in this case, the acid binding agent used herein may include heterocyclic amines such as pyridine and dimethylaminopyridine, and aliphatic amines such as trimethylamine, triethylamine and Diisopropylamine; preferred aliphatic amines (especially Diisopropylamine).

Used here disubstituted alkoxyamine may include phosphine derivatives such as bis(diisopropylamino)canadaceftin, bis(diethylamino)methanesulfonylaminoethyl, bis(diisopropylamino)(2,2,2-trichloroethane)phosphine and bis(diisopropylamino)(4 chlorpheniramine)phosphine, preferably bis(diisopropylamino)canadaceftin.

When used disubstituted alkoxyamine, apply the acid, and in this case, the acid used preferably may include tetrazole, acetic acid or p-toluensulfonate acid.

The reaction temperature is not limited, but usually it ranges from 0 to 80

After the interaction, the desired compound (16) this reaction receive, for example, by appropriate neutralization of the reaction mixture, removing insoluble substances by filtration, if they are present, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent. Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization, re-precipitation or chromatography, etc.,

Alternative this stage is intended to obtain compound (16) by reacting compound (15) obtained in Stage E-1, Tris-(1,2,4-triazolyl)postiton in an inert solvent, preferably halogenated hydrocarbons, such as methylene chloride, followed by addition of water to cause N-Vospominanie.

The reaction temperature concreted src="https://img.russianpatents.com/chr/176.gif">C.

The time of interaction varies depending on the starting material, reagent and reaction temperature, but it usually ranges from 5 min to 30 h, preferably 30 min, when the reaction is performed at room temperature.

After the interaction, the desired compound (16) this reaction receive, for example, by appropriate neutralization of the reaction mixture, removing insoluble substances by filtration, if they are present, adding an organic solvent that is not miscible with water such as ethyl acetate, washing with water, separation of the organic layer containing the desired compound, drying over anhydrous magnesium sulfate and distillation of the solvent. Thus obtained target product can be further purified, if necessary, in the traditional way, for example by recrystallization, re-precipitation or chromatography, etc.,

Stage E-3

At this stage the target oligonucleotide analogue obtained using the automatic DNA sintetizzatore using at least one compound (16) obtained in stage E-2, and a commercially available fostamatinib reagents necessary for obtaining oligonucleotide analogue with VC is the dialogue, having the desired nucleotide sequence can be synthesized using DNA sintetizzatore, such as Perkin-Elmer, model 392, using phospholidine method, in accordance with the method described in the literature (Nucleic Acids Research, 12 so, S. 4539 (1984)).

In addition, in the case of transformation in tioat wish tiata derivative can be obtained according to the method described in literature (Tetrahedron Letters, 32 so, 3005 S. (1991), J. Am. Chem. Soc., so 112, S. 1253 (1990)), using, in addition to sulfur reagent, which forms tioat, by interacting with the trivalent phosphoric acid, such as tetraethylsilane (TETD, the company Applied Biosystems Inc.) or Beaucage reagent (firm Millipore Corp.).

The obtained oligonucleotide analogue can be purified using column OligoPak (chromatographic column with reversed phase), and the purity of the product can be confirmed by using HPLC (WAIK).

Chain length obtained oligonucleotide analogue is usually from 2 to 50, preferably from 10 to 30 nucleotides.

The ability to form complementary chain and resistance to nucleases derived oligonucleotide analogue can be determined in accordance with methods described below.

Test method 1

Spoke RNA can be determined by different annealing synthesized oligonucleotide analogues with oligonucleotide analogues consisting of DNA or RNA of natural origin having a complementary sequence, and measuring the melting temperature (value TM).

The sample solution containing equal amounts of oligonucleotide analogue and a complementary oligonucleotide natural origin in nutrifaster buffer solution, placed in a boiling water bath and then slowly cooled down to room temperature within a certain time (annealing). Then the solution is placed in the cuvette of a spectrophotometer (for example, Shimadzu UV-2100PC), gradually increase the temperature from 20 to 90C, followed by measuring the absorption at 260 nm.

Test method 2. Determination of resistance to nucleases

To the oligonucleotide in a buffer solution add nuclease and the mixture is heated. In the example used fosfodiesterazu snake venom, endonuclease P1 and S1 endonuclease. Testing can be done at any buffer solution, however, it is preferable to use a buffer solution suitable for the specific enzyme; so if fosfodiesterasa snake venom using Tris-HCl and in the case of P1 endonuclease using sodium acetate. In addition, if necessary, a buffer solution is added ions METAR> in the case of endonucleases. The reaction temperature is preferably from 0 to 100S, more preferably from 30 to 50C.

After a certain period of time add ethylenediaminetetraacetic acid (EDTA) and the mixture is heated for 2 minutes at 100To stop the reaction.

To estimate the remainder of the oligonucleotide using radioisotope-labeled oligonucleotide with the subsequent analysis of the product of the cleavage reaction using any of the analyzer, such as HPLC or by staining dye (such as ethidiumbromid) analysis using the computer.

Dosage forms oligonucleotide analogue having one, or two or more structures of the formula (2) of the present invention can be tablets, capsules, granules, powders or syrups for oral administration, or injections, or suppositories for parenteral use. Such dosage forms get well-known methods using additives such as fillers (for example, organic fillers, such as derivatives of sugars, such as lactose, sucrose, glucose, mannitol and sorbitol; starch derivatives such as the crystalline cellulose; Arabian gum; dextran; and Pullulan; and inorganic fillers such as derivatives of silicates, for example, light silicic anhydride, synthetic aluminum silicate, calcium silicate and metasilicate of magnesium aluminate; phosphates such as calcium phosphate; carbonates such as calcium carbonate; and sulfates, such as calcium sulfate), lubricants (e.g. stearic acid, metal salts of stearic acid such as calcium stearate and magnesium stearate; talc; colloidal silica; waxes such as beeswax and spermaceti; boric acid; adipic acid; sulfates, such as sodium sulfate; glycol; fumaric acid; sodium benzoate; DL-leucine; sodium salts of fatty acids; laurilsulfate, such as sodium lauryl sulfate and lauryl sulfate, magnesium; silicic acids such as silicic anhydride and silicic acid hydrate; and the above starch derivatives), binders (for example, hydroxypropylcellulose, hypromellose, polyvinylpyrrolidone, Macrogol, and compounds similar to the above fillers), shredders (for example, cellulose derivatives such as nizkozameshhennoj hydroxypropylcellulose, carboxymethylcellulose, callicarpenal is lulzy, such as carboximetilkrahmal, carboximetilkrahmal sodium and bridge polyvinylpyrrolidone), preservatives (pair-oxybenzoates, such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenethyl alcohol; benzylaniline; derivatives of phenol such as phenol and cresol; thimerosal; dehydroacetic acid; and sorbic acid), corrective additives (for example, the commonly used sweeteners, acidifying agents, flavorings and other), thinners, etc.

Although these forms may vary depending on the state of the disease, age of patient, application methods, and so on, for example, in the case of oral administration, it is desirable to introduce the active ingredient in an amount of from 0.01 mg/kg body weight (preferably 0.1 mg/kg body weight) up to 1000 mg/kg body weight (preferably 100 mg/kg body weight) and in the case of intravenous purpose, it is desirable to introduce the active ingredient in an amount of from 0.001 mg/kg body weight (preferably 0.01 mg/kg of body weight) up to 100 mg/kg body weight (preferably 10 mg/kg body weight), in a single daily dose or in the form of doses distributed to reception several times a day, respectively.

In NMR spectra: Shir. - wide, s - singlet, d - doublet is as fast atoms); UV - UV range.

INFORMATION CONFIRMING the POSSIBILITY of carrying out the INVENTION

Example 1. 3',5'-di-O-Benzyl-2'-O,4'-C-ethylene-4-N-benzoylation (number of illustrative compounds 2-34)

Added 2N aqueous solution of sodium hydroxide (68 ml) solution of the compound obtained in comparative example 11 (6,80 g, 8,86 mmol) in pyridine (136 ml) at 0C and the mixture is stirred at room temperature for 1 hour. The reaction mixture is neutralized by adding dropwise the aqueous 20% acetic acid, and extracted with chloroform. Washed chloroformate layer saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 100:3 as eluent) to obtain specified in the title compound (3.3 grams, of 6.02 mmol, 68%).

An NMR spectrum1H (400 MHz, Dl3): 8,64(2H, Shir. C), 7,89 (2H, d, a 7.6 Hz), 7, 64-7, 60 (1H, m), 7,54-7,51 (2H, m), 7, 48-7, 37 (3H, m), of 7.36-7,26 (8H, m), 6,18 (1H,s), 4,70 (1H, d, 11 Hz), 4,60 (1H, d, 11 Hz), 4,55(1H, d, 11 Hz), 4,46 (1H, d, 2,9 Hz), was 4.42 (1H, d, 11 Hz), 4,10-was 4.02 (2H, m) to 3.89(1H, d, 2,9 Hz in), 3.75(1H, d, 11 Hz), 3,62 (1H, d, Hz), 2,34-of 2.26 (1H, m), 1,39-of 1.36 (1H, m).

Mass spectrum (fast atom bombardment) FAB-MAS (mNBA): 554 (M+N)+.

Example 2. 2'-O,4'-C-Ethyl is ethane added dropwise into a solution of the compound, obtained in example 1 (e 2.06 g, 3.72 mmol) in anhydrous methylene chloride (317 ml) at -78C, and the mixture is stirred at this temperature for 1 hour. The reaction mixture is slowly warmed to -20C, the reaction flask is placed in a bath of ice and sodium chloride and the mixture is stirred at a temperature between -20 and -10C for 2 hours. Slowly add in the mixture of methanol (12 ml) and the mixture is stirred for 10 minutes, the pH Value of the reaction mixture is set to 7-8 by adding dropwise a saturated aqueous solution of sodium bicarbonate. The mixture is heated to room temperature and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 100:5 as eluent) to obtain the specified header connection (1,21 g, 3,24 mmol, 87%) as a white solid.

An NMR spectrum1H (500 MHz, DMSO-d6): 11,23 (lH, Shir. C) to 8.70 (1H, d, 7.2 Hz), 8,00 (2H, q, j 7.5 Hz), 7,3-6 (4H, m), 5,97 (1H, s), to 5.35 (1H, DD, 5 and 10 Hz), 4,10 (1H, DD, 5 and 10 Hz), a 4.03 (1H, d, 3.2 Hz), 3.95 to of 3.85 (2H, m) a 3.83 (1H, d, 3.2 Hz), 3,65-3,51 (2H, m), 2.06 to to 1.98 (1H, m) of 1.26 (1H, m).

FAB-MAS (mNBA): 374 (M+N)+.

Example 3. 2'-O,4'-C-Atransition (number of illustrative compound 2-3)

Rast is giving night. The mixture is concentrated to dryness in order to obtain the specified header connection (0,054 g, 75%) as a white solid.

An NMR spectrum1H (500 MHz, DMSO-d6): 8,18 (1H, q, j 7.4 Hz), 7,10 (2N, W), of 5.84 (1H, s) 5,69 (1H, d, and 7.6 Hz), 5,27-5,24 (2H, m), 3,86 (1H, d, 3.2 Hz), 3, 90-3, 78 (2H, m), 3,76 (1H, d, 3.2 Hz), of 3.56 (1H, DD, 5.5 and 12 Hz), 3,49 (1H, DD, 5, 5 and 12 Hz), 2,01-of 1.93 (1H, dt, 7.5 and 12 Hz), 1,22 (1H, DD, 3.6 and 13 Hz).

FAB-MAS (mNBA): 270(M+N)+.

Example 4. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation

(number of illustrative compounds 2-39)

A solution of the compound obtained in example 2 (1.29 g, 3.46 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous pyridine (26 ml) under nitrogen atmosphere and added to a solution of 4,4'-dimethoxytrityl (1,76 g, 5.18 mmol) and the mixture stirred at room temperature overnight. To the reaction mixture is added a small amount of methanol and then the solvent is evaporated in vacuum. The residue is distributed between water and chloroform, and the organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel (using cm,10 g, 3.11 mmol, 90%) as a colorless amorphous solid.

An NMR spectrum1H (270 MHz, DMSO-d6): 11,27 (1H, Shir. C) 8,59 (1H, m), 6,92-8, 01 (N, m), 6,03 (1H, s) to 5.56 (1H, m), 4,17 (1H, m), 4,08 (1H, m), 3,86 (2H, m), of 3.77 (6N, (C), 3,24 (2H, m) to 1.98 (1H, m), 1,24 (1H, m).

FAB-MAS (mNBA): 676 (M+N)+.

Example 5. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (number of illustrative compounds 2-235)

A solution of the compound obtained in example 4 (6,53 g to 9.66 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved under nitrogen atmosphere in anhydrous dichloromethane (142 ml). To the solution was added N,N-Diisopropylamine (2,80 ml, at 16.1 mmol) and then 2-Tianeti-N,N-diisopropylchlorophosphoramidite (2,16 ml, to 9.66 mmol) is added dropwise in the bath with ice. The mixture is stirred at room temperature for 6 hours. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : triethylamine = 50:1 - dichloromethane : ethyl acetate : triethylamine = 60: 30: 1 as eluent) to obtain specified in the header of the connection is), of 1.35(1H, m), 2,11 (1H, m), 2,3 (2H, m), 3,35-3,7 (6N, m), 3,8(6N, m), 3,9-4,1 (2H, m) to 4.33 (1H, m), of 4.45 (1H, m), 6,23 (1H, s), 6,9 (4H, m), 7.3 to 7.9 in (15 NM, m) of 8.7 to 8.8 (1H, m).

Example 6. 3',5'-Di-O-benzyl-2'-O,4'-C-ethylene-5-methyluridine (number of illustrative compound 2-22)

An aqueous solution (2N) of sodium hydroxide and a mixed solution (5 ml) specified in the mixed solution has a composition pyridine : methanol : water = 65:30:5, added to the compound obtained in comparative example 10 (418 mg, of 0.62 mmol) in a mixture of pyridine : methanol : water = 65: 30: 5 (5 ml) at 0C, and the mixture is stirred at room temperature for 15 minutes the Reaction mixture is neutralized 1N hydrochloric acid and extracted with ethyl acetate (approximately 30 ml). The organic layer was washed with saturated aqueous sodium bicarbonate (approximately 30 ml) and saturated aqueous sodium chloride (approximately 30 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of hexane : ethyl acetate = 1:1 as eluent) to obtain a colorless amorphous solid (228 mg, 0.49 mmol, 79%).

An NMR spectrum1H (400 MHz, Dl3): 1,35 (1H, d, 13 Hz), of 1.41 (3H, s), 2,28 (1H, dt, 9,4 and 13 Hz), 3,60 (1H, d, 11 Hz), 3,76 ,75 (1H, d, 12 Hz), the 6.06 (1H, s), and 7.3 (10H, m), to $ 7.91 (1H, s), 8,42 (1H, Shir. C).

FAB-MAS (mNBA): 465 (M+H)+.

Example 7. 2'-O,4'-C-Ethylene-5-methyluridine (number of illustrative compound 2-2)

A solution of the compound obtained in example 6 (195 mg, 0.42 mmol), in methanol (10 ml) stirred in an environment of hydrogen at atmospheric pressure for 5 hours. The reaction mixture is filtered to remove catalyst and the filtrate concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane: methanol =10:1 as eluent) to obtain a colorless powder (76 mg, 0,268 mmol, 64%).

An NMR spectrum1H (400 MHz, CD3OD): 1,33 (1H, DD, 3,8 and 13 Hz), to 1.86 (3H, d, and 0.9 Hz), was 1.94 (1H, DDD, 7,5, 11.7 and 13 Hz), 3,68 (1H, d, 12 Hz in), 3.75 (1H, d, 12 Hz), 3,9-4,0 (2H, m), of 4.05(1H, d, 3.2 Hz), 4.09 to (1H, d, 3.2 Hz), 6,00 (1H, s), of 8.28 (1H, d, 1,1 Hz).

FAB-MAS (mNBA): 285(M+N)+.

Example 8. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-5-methyluridine (number of illustrative compound 2-27)

A solution of the compound obtained in example 7 (1.45 g, 5,10 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous pyridine (44 ml) under nitrogen atmosphere and the solution was added 4,4'-dimethoxytrityl (2,59 g of 7.65 mmol) and the mixture stirred praetorial is evaporated in vacuum. The residue is distributed between water and chloroform, and the organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 100:10 as eluent) to obtain the specified header connection (2,42 g of 4.13 mmol, 81%) as a colorless amorphous solid.

An NMR spectrum1H (270 MHz, DMSO-d6): 11,36 (1H, s), to 7.68 (1H, s), 6.90 to-7,44 (13H, m), of 5.89 (1H, s), of 5.55 (1H, d), 4.09 to (1H, m), Android 4.04 (1H, d), 3,82 (2H, m), 3,74 (6N, (C), 3,19 (2H, m), 1,99 (1H, m) of 1.36 (1H, m) of 1.17 (3H, C).

FAB-MAS (mNBA): 587 (M+N)+.

Example 9. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-5-methyluridine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (number of illustrative compounds 2-234)

A solution of the compound obtained in example 8 (4.72 in g, with 8.05 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved under nitrogen atmosphere in anhydrous dichloromethane (142 ml). Add N,N-Diisopropylamine (2,80 ml, at 16.1 mmol) in a solution and then added dropwise 2-Tianeti-N,N-diisopropylchlorophosphoramidite (2,16 ml, to 9.66 mmol) in an ice bath. The mixture is stirred at room temperature in those who m solution of sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of hexane : ethyl acetate : triethylamine = 50:50:1 - hexane : ethyl acetate: triethylamine = 30: 60: 1 as eluent) to obtain the specified header connection (5,64 g, 7,17 mmol, 89%) as a colorless amorphous solid.

An NMR spectrum1H (400 MHz, Dl3): 1.1 to 1.2 (15 NM, m), 1,4 (1H, m), of 2.08 (1H, m), 2,4 (2H, m), 3,2-4,0 (14N, m), to 4.38 (2H, m), 4,47 (1H, m), the 6.06 (1H, s), 6.8 or 6.9 (4H, m), 7,2-7,5 (N, m), to $ 7.91 (1H, m).

FAB-MAS (mNBA): 787 (M+H)+.

Example 10. 3', 5'-Di-O-benzyl-2'-O,4'-C-ethylene-6-N-benzyladenine

(number of illustrative compound 1-23)

Aqueous 2N solution of sodium hydroxide and a mixed solution (5 ml) indicated mixed solution contains a mixture of pyridine : methanol : water = 65:30:5, added to the compound obtained in comparative example 12 (238 mg, 0.30 mmol) in a mixture of pyridine : methanol : water = 65:30:5 (5 ml) at 0C and the mixture is stirred at room temperature for 15 minutes the Reaction mixture is neutralized 1N hydrochloric acid and extracted with ethyl acetate (approximately 30 ml).

The organic layer was washed with saturated aqueous sodium bicarbonate (approximately 30 ml) and saturated aqueous sodium chloride (about whom and on a column of silica gel (using a mixture of dichloromethane : methanol = 50:1 as eluent) to to obtain a colorless amorphous solid (133 mg, 0.23 mmol, 78%).

An NMR spectrum1H (400 MHz, CDCl3): the 1.44 (1H, d, 13 Hz), 2,31 (1H, DD, 13 and 19 Hz), of 3.56 (1H, d, 11 Hz), 3,70 (1H, d, 11 Hz), 4,10 (2H, m), 4,24 (1H, s), of 4.45 (1H, d, 12 Hz), 4.53-in-4,67 (4H, m), of 6.52 (1H, s), and 7.3 (10H, m), 7,53 (2H, m), a 7.62 (1H, m), 8,03 (2H, d, a 7.6 Hz), 8,66 (1H, s), 8,78 (1H, s), of 9.00 (1H, Shir. C).

FAB-MAS(mNBA): 578(M+N)+.

Example 11. 2'-O,4'-C-Ethylene-6-N-benzyladenine (number of illustrative compound 1-178)

Slowly added dropwise 1M solution of trichloride boron (1.5 ml, 1.5 mmol) in dichloromethane to a solution of the compound obtained in example 10 (116 mg, 0.20 mmol) in anhydrous methylene chloride (5 ml) at -78C and the mixture is stirred at this temperature for 3 hours. To the reaction mixture are added 1M solution of trichloride boron (1.5 ml, 1.5 mmol) in dichloromethane and the mixture is stirred for 2 hours. The mixture is slowly warmed to room temperature, and then rapidly cooled to -78C and then added to methanol (5 ml). The reaction mixture is slowly warmed to room temperature and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 9:1 as eluent) to obtain white powder coated examination bed with d, 12 Hz), Android 4.04 (1H, DD, 7.3 and 12 Hz), 4,15 (1H, dt, of 4.3 and 9.4 Hz), 4,36 (1H, d, 3.2 Hz), 4,43 (1H, d, 3.2 Hz), to 6.57 (1H, s), EUR 7.57 (2H, m), 7,66 (1H, m), of 8.09 (2H, d, 8.0 Hz), 8,72 (1H, s), cent to 8.85 (1H, s).

FAB-MAS (mNBA): 398 (M+N)+.

Example 12. 2.' -O,4'-C-Utilizationin (number of illustrative compounds 1-7)

A solution of the compound obtained in example 11 (14 mg, 0.035 mmol) in methanol saturated with ammonia (1 ml), incubated over night. The mixture is concentrated and the residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol =10:1 as eluent) to give a white powder (10 mg, 0,034 mmol, 98%).

An NMR spectrum1H(400 MHz, CD3OD): 1,32 (1H, DD, 4 and 13 Hz), 2,04 (1H, dt, 7.4 and 12 Hz), 3,53 (1H, DD, 5 and 12 Hz), 3,61 (1H, DD, 5,2 and 12 Hz), 3,90 (1H, DD, 7.4 and 12 Hz), of 3.97 (1H, dt, 4 and 12 Hz), is 4.15 (1H, q, j 3.1 Hz), is 4.21 (1H, q, j 3.1 Hz), 5,27 (1H, t, 5,2 Hz), of 5.39 (1H, q, j 3.1 Hz), 6,33 (1H, s), 7,29 (2H, s), 7,66 (1H, m) to 8.14(1H, s), 8,42(1H, s).

FAB-MAS (mNBA): 294(M+N)+.

UV-spectrum (max): 260 (m), 260 (pH 1), 258 (rn).

Example 13. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-6-N - benzyladenine

(number of illustrative compound 1-31)

A solution of the compound obtained in example 11 (14 mg, 0.035 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous pyridine (1 ml) Armenie 5 hours. To the reaction mixture is added a small amount of methanol, and then the solvent is evaporated in vacuum. The residue is distributed between water and chloroform, the organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol =100:5 as eluent) to obtain specified in the title compound (18 mg, was 0.026 mmol, 73%) as a colorless amorphous solid.

An NMR spectrum1H (400 MHz, Dl3): and 1.63 (1H, m), and 2.14 (1H, 7,5,12 and 13 Hz), 3, 37 (1H, d, 11 Hz), to 3.41 (1H, d, 11 Hz), 3,79 (6N, (C), 4,10 (2H, m), 4,48 (1H, d, 3.3 Hz), 4,59 (1H, d, 3.3 Hz), is 6.54 (1H, s), 6,85 (4H, m), 7,2-7,6 (N, m), 8,02 (2H, m), to 8.45 (1H, s), 8,82 (1H, s), of 9.02 (1H, Shir. C).

FAB-MAS (mNBA): 700 (M+N)+.

Example 14. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide

(number of illustrative compounds 1-186)

A solution of the compound obtained in example 13 (16 mg, is 0.023 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous dichloromethane (0.5 ml) in nitrogen atmosphere. Tetrazolium salt of N,N-Diisopropylamine (10 mg) like 20 ml) in an ice bath. The mixture is stirred at room temperature overnight. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : ethyl acetate = 2:1 as eluent) to obtain specified in the title compound (20 mg, of 0.022 mmol, 97%) as a white solid.

An NMR spectrum1H (400 MHz, Dl3): 1,0-1,2 (N, m), and 1.54 (1H, m) to 2.15 (1H, m), 2,33 (2H, m), 3,3-3,6 (6N, m), 3,80 (6N, C) 4,08 (2H, m) and 4.65 (1H, m), and 4.75 (1H, m), 6,53 (1H, s), at 6.84 (4H, m), 7,2-7,6 (N, m), 8,01 (2H, m), 8,53 (1H, s), 8,83(1H, s), 9,01 (1H, Shir. C).

FAB-MAS(mNBA): 900(M+H)+.

Example 15. 3',5'-Di-O-benzyl-2'-O,4'-C-ethylenedi (number of illustrative compounds 2-10)

Aqueous 1N solution of sodium hydroxide (2 ml) are added to a solution of the compound obtained in comparative example 13 (194 mg, 0,292 mmol) in pyridine (3 ml) at 0C and the mixture is stirred at room temperature for 30 minutes, the Reaction mixture is neutralized 1N hydrochloric acid and extracted with ethyl acetate (10 ml). The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous chloride nutri column of silica gel using a mixture of dichloromethane : methanol=100:3 as eluent), receiving a colorless oil (105 mg, 0,233 mmol, 80%).

An NMR spectrum1H (400 MHz, CDCl3): of 1.36 (1H, m) to 2.29 (1H, m), 3,63 (1H, d, 11 Hz), 3,74 (1H, d, 11 Hz), a 3.87 (1H, d, 2,9 Hz), a 4.03 (2H, m), the 4.29 (1H, d, 2,9 Hz), 4,49 (1H, d, 12 Hz), 4,50 (1H, d, 11 Hz), a 4.53 (1H, d, 11 Hz), to 4.73 (1H, d, 12 Hz), 5,20 (1H, DD, 2 and 8 Hz), 6,04 (1H, s), 7,2-7,4 (10H, m), 8,13 (1H, d, 8.2 Hz), to 8.57 (1H, Shir. C).

FAB-MAS (mNBA): 451 (M+N)+.

Example 16. 2'-O,4'-C-Acylceramide (number of illustrative compound 2-1)

A solution of the compound obtained in example 15 (100 mg, 0,222 mmol), in methanol (4 ml) stirred in an environment of hydrogen at atmospheric pressure in the presence of catalytic hydrogenation over 5 hours. The reaction mixture is filtered to remove the catalyst, and the filtrate was concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 10: 1 as eluent) to obtain a colorless oil (45 mg, 0,167 mmol, 75%).

An NMR spectrum1H (400 MHz, CD3OD): 1,35 (1H, DD, 4 and 13 Hz), 2,13 (1H, DDD, 7, 11 and 13 Hz), 3,66 (1H, d, 12 Hz), to 3.73 (1H, d, 12 Hz), 3,91-4,08 (2H, m) to 4.01 (1H,d, 3.2 Hz), 4,12(1H, d, 3.2 Hz), to 5.66 (1H, d, 8.2 Hz), 6,00 (1H, s) of 8.37 (1H, d, 8,2 Hz).

FAB-MAS (mNBA): 271 (M+H)+.

Example 17. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylenedi (number of illustrative compound 2-15)

Solution connection, receive the ing water. The product is dissolved in anhydrous pyridine (3 ml) under nitrogen atmosphere, is added to the solution of 4,4'-dimethoxytrityl (50 mg, 0.15 mmol) and the mixture is stirred at room temperature overnight. To the reaction mixture is added a small amount of methanol and then the solvent is evaporated in vacuum. The residue is distributed between water and chloroform, the organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 100:3 as eluent) to obtain specified in the title compound (25 mg, 0,044 mmol, 42%) as a colourless oil.

An NMR spectrum1H (400 MHz, CD3OD): 1,35 (1H, DD, 3 and 14 Hz), 2,03 (1H, DDD, 8, 11 and 14 Hz), 2,46 (1H, d, 8 Hz), to 3.36 (1H, d, 11 Hz), to 3.41 (1H, d, 11 Hz), 3,80 (3H, s), 3,81 (3H, s), of 3.97 (2H, m), is 4.21 (1H, d, 3.2 Hz), 4,33 (1H, Shir. m), 5,31 (1H, m), 6,10 (1H, s) 6,86 (4H, m), 7,2-7,5 (N, m) of 8.27 (1H, d, 8.2 Hz), 8,43 (1H, Shir. C).

FAB-MAS (mNBA): 573(M+H)+.

Example 18. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylenedi-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (number of illustrative compounds 2-233)

A solution of the compound obtained in example 17 (6 mg, 0,0105 mmol) in anhydrous Piri is) in nitrogen atmosphere. Tetrazolium salt of N,N-Diisopropylamine (3 mg) is added to the solution and then added dropwise 2-Tianeti-N,N,N',N'-tetraisopropylphosphoramidite (approximately 5 ml) in an ice bath. The mixture is stirred at room temperature overnight. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : ethyl acetate = 2:1 as eluent) to obtain specified in the title compound (8 mg) as a white solid.

An NMR spectrum1H (400 MHz, CDCl3): 1.1 to 1.2 (13H, m), is 2.09 (1H, m), 2,4 (2H, m), 3,3-3,6 (6N, m), 3,81 (6N, m), of 3.94 (2H, m), 4,35 (1H, m), 4,47 (1H, m), is 5.18 (1H, d, 8.2 Hz), between 6.08 (1H, s) 6,86 (4H, m), 7,2-7,4 (N, m), 8,31 (1H, d, 8,2 Hz).

FAB-MAS (mNBA): 773(M+H)+.

Example 19. 3',5'-Di-O-benzyl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine (number of illustrative compounds 2-46)

Aqueous 1N solution of sodium hydroxide (5 ml) are added to a solution of the compound obtained in comparative example 14 (310 mg, 0,396 mmol), in pyridine (5 ml) at 0C and the mixture is stirred at room temperature for 20 minutes the Reaction mixture is neutralized by adding to the n aqueous solution of sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 100:2 as eluent) to obtain specified in the title compound (190 mg, 0,334 mmol, 84%).

An NMR spectrum1H (400 MHz, CDCl3): of 1.37 (1H, m), 1,58 (3H, s), is 2.30 (1H, dt, 10 and 13 Hz), to 3.64 (1H, d, 11 Hz), with 3.79 (1H, d, 11 Hz), 3,95 (1H, d, 3.0 Hz), Android 4.04 (2H, DD, 2,3 and 10 Hz), 4,37 (1H, d, 3.0 Hz), 4,50 (1H, d, 12 Hz), 4,56 (1H, d, 11 Hz), br4.61 (1H, d, 11 Hz), was 4.76 (1H, d, 12 Hz), 6,11 (1H, s), 7,2-7,5 (13H, m), of 8.09 (1H, s), 8,29 (2H, m).

FAB-MAS (mNBA): 568(M+H)+.

Example 20. 2'-O,4'-C-Ethylene-4-N-benzoyl-5-methylcytidine (number of illustrative compounds 2-226)

Added dropwise 1M solution of trichloride boron (1.6 ml) in dichloromethane to a solution of the compound obtained in example 19 (120 mg, 0,211 mmol) in anhydrous dichloromethane (5 ml) at a temperature of -78C and the mixture was stirred at -78With 4 hours. Slowly added dropwise methanol (1 ml) to the mixture and the mixture is stirred for 10 minutes, the pH Value of the reaction mixture was adjusted to 7-8 by adding dropwise a saturated aqueous solution of sodium bicarbonate. The reaction mixture is heated to room temperature and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : meta is solid substances.

An NMR spectrum1H (400 MHz, DMSO-d6): 1,24 (1H, m), a 2.01 (3H, s), 2,0 (1H, m), of 3.54 (1H, DD, 5.4 and 12 Hz), to 3.64 (1H, DD, 5.4 and 12 Hz), 3,88 (3H, m, 4,10 (1H, m), are 5.36 (1H, q, j 5.4 Hz), 5,49 (1H, t, 5.0 Hz), 5,95 (1H, s), 7,4-7,6 (3H, m), 8,21 (2H, m), 8,49 (1H, s), 13,17 (1H, Shir. C).

FAB-MAS (mNBA): 388 (M+N)+.

Example 21. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine (number of illustrative compounds 2-51)

A solution of the compound obtained in example 20 (44 mg, 0,114 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous pyridine (1 ml) under nitrogen atmosphere, is added to the solution of 4,4'-dimethoxytrityl (60 mg, 0,177 mmol) and the mixture is stirred at room temperature overnight. To the reaction mixture is added a small amount of methanol and then the solvent is evaporated in vacuum. The residue is distributed between water and chloroform. The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol = 100:4 as eluent) to obtain specified in the title compound (73 mg, 0,106 mmol, 93%) as a colourless mA is Hz), of 3.80 (3H, s), 3,81 (3H, s) to 3.99 (2H, m), 4,30 (1H, d, 3.3 Hz), 4,39 (1H, m), 6,12 (1H, s), 6,85 (4H, m), 7,2-7,5 (N, m), 8,03 (1H, s), of 8.28 (2H, m).

FAB-MAS (mNBA): 573 (M+N)+.

Example 22. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (number of illustrative compounds 2-236)

A solution of the compound obtained in example 21 (35 mg, 0,0507 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous dichloromethane (1 ml) under nitrogen atmosphere. Tetrazolium salt of N,N-Diisopropylamine (17 mg) is added to the solution and then added dropwise 2-Tianeti-N,N,N',N'-tetraisopropylphosphoramidite (32 μl, 0.1 mmol) in a bath of ice. The mixture is stirred at room temperature overnight. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : ethyl acetate= 2:1 as eluent) to obtain specified in the title compound (40 mg, 0,0445 mmol, 89%) as a white solid.

An NMR spectrum1H (400 MHz, Dl3): 1.1 to 1.2 (N, m) of 1.36 (3H, s) to 1.37 (1H, m), 2,10 (1H, m), a 2.36 (2H, m), 3,3-3,6 (6N, m),

Example 23. 2'-O,4'-C-Ethylene-5-methylcytidine (number of illustrative compounds 2-226)

A solution of the compound obtained in example 20 (11.6 mg 0,030 mmol) in methanol saturated with ammonia (2 ml), incubated over night. The mixture is concentrated to obtain a white solid (8.5 mg, 0,030 mmol).

An NMR spectrum1H (400 MHz, DMSO-d6): 1,20 (1H, m), equal to 1.82 (3H, s) of 1.97 (1H, m), 3,49 (1H, DD, 5 and 12 Hz), to 3.58 (1H, DD, 5 and 12 Hz), 3,85 (2H, m), 5,23 (1H, d, 5 Hz), 5,32 (1H, t, 5 Hz), of 5.84 (1H, s), 6,7 (1H, Shir. C) to 7.2 (1H, Shir. C) 8,08 (1H,s).

FAB-MAS (mNBA): 284 (M+N)+.

UV(max): 279 (rn), 289 (pH1), 279 (rn).

Example 24. 3',5'-Di-O-benzyl-2'-O,4'-C-ethylene-2-N-isobutylamine (number of illustrative compound 1-24)

Aqueous 1N solution of sodium hydroxide (2 ml) are added to a solution of the compound obtained in comparative example 15 (approximately 200 mg) in pyridine (2 ml) and the mixture is stirred at room temperature for 15 minutes the Reaction mixture is neutralized 1N hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on a column with silicate substance (20 mg, being 0.036 mmol, 6%, stage 2).

An NMR spectrum1H (400 MHz, CDCl3): l,27 (3H, c), of 1.29 (3H, s), 1,43 (1H, DD, 3 and 13 Hz), 2,28 (1H, m), 2,59 (1H, qui, 6.9 Hz), 3,54 (1H, d, 11 Hz), 3,68 (1H, d, 11 Hz), a 4.03 (2H, m), is 4.15 (1H, q, j 3.0 Hz), or 4.31 (1H, d, 3.0 Hz), of 4.45 (1H, d, 12 Hz), 4,56 (1H, d, 12 Hz), br4.61 (1H, d, 12 Hz), 4,63 (1H, d, 12 Hz), 6,18 (1H, s), 7,2-7,4 (10H, m), 8,19 (1H, s), 11,93 (1H, Shir. C).

FAB-MAS (mNBA): 560 (M+N)+.

Example 25. 2'-O,4'-C-Ethylene-2-N-isobutylamine (number of illustrative compound 1-177)

A solution of the compound obtained in example 24 (10 mg, 0.018 mmol), in methanol (2 ml) is stirred in an environment of hydrogen at atmospheric pressure for 5 hours. The reaction mixture is filtered to remove the catalyst, and the filtrate was concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol =10:2 as eluent) to obtain a colorless oil (5 mg, of 0.013 mmol, 72%).

An NMR spectrum1H (400 MHz, CD3OD): to 1.21 (3H, s) to 1.22 (3H, s) of 1.41 (1H, DD, 4 and 13 Hz), to 2.18 (1H, m), 2,69 (1H, qui, 6.9 Hz), of 3.69 (1H, d, 12 Hz), 3,76 (1H, d, 12 Hz), 4,0 (2H, m), 4.26 deaths (1H, d, 3.2 Hz), 4,30 (1H, d, 3.2 Hz), 6,30 (1H, s), to 8.40 (1H, s).

FAB-MAS (mNBA): 380(M+H)+.

Example 26. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamine (number of illustrative compound 1-35)

A solution of the compound obtained in example 25 (5 mg, 0,013 the bottom pyridine (1 ml) in nitrogen atmosphere, to the solution was added 4,4'-dimethoxytrityl (14 mg, 0.04 mmol) and the mixture is stirred at 40 ° C for 3 hours. To the reaction mixture is added a small amount of methanol and then the solvent is evaporated in vacuum. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : methanol =100:6 as eluent) to obtain specified in the title compound (4 mg, 0,0059 mmol, 45%) as a colourless solid.

An NMR spectrum1H (400 MHz, Dl3): of 1.26 (3H, d, a 1.4 Hz), of 1.28 (3H, d, a 1.4 Hz), of 1.66 (1H, m) to 2.15 (1H, m), 2,59 (1H, qui, 6.9 Hz), the 3.65 (1H, m), of 3.78 (1H, m) 4,06 (2H, m), 4,35 (1H, m), to 4.38 (1H, d, 3.2 Hz), 6,23 (1H, s), 6,8 (4H, m), 7,2-7,5 (N, m), 8,01 (1H, s), 8,19 (1H, Shir. C).

FAB-MAS (mNBA): 682 (M+N)+.

Example 27. 5'-O-Dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamino-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide (number of illustrative compound 1-185)

A solution of the compound obtained in example 26 (4 mg, 0,0058 mmol) in anhydrous pyridine is subjected to azeotropic distillation to remove water. The product is dissolved in anhydrous dichloromethane (0.5 ml) in nitrogen atmosphere. Tetrazolium salt of N,N-Diisopropylamine (5 mg) are added to the solution and then added dropwise 2-Tianeti-N,N,N',N'-tetraisopropylphosphoramidite washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride and concentrated in vacuo. The residue is purified chromatographically on a column of silica gel using a mixture of dichloromethane : ethyl acetate = 2:1 as eluent) to obtain specified in the title compound (4 mg) as a white solid.

An NMR spectrum1H (400 MHz, CDCl3): 1.1 to 1.4 (N, m) 2,1 (1H, m), 2,4 (2H, m), 2,6 (1H, m), 3,3-3,6 (6N, m), 3,8 (6N, s), 4,0-4,6 (4H, m), 6,2 (1H, s), 6,8 (4H, m), 7,2-7,5 (N, m) and 8.1 (1H, s).

Example 28. 2'-O,4'-C-Teleguinin (number of illustrative compounds 1-5)

A solution of the compound obtained in example 25 (0.5 mg) in methanol saturated with ammonia (0.5 ml), incubated at 60C for 5 hours. The mixture was concentrated, obtaining a white powder (0.4 mg).

FAB-MAS (mNBA): 310 (M+N)+.

UV(max): 255 (m), 256 (pH 1), 258-266(rn).

Example 29. Synthesis of oligonucleotide derivatives

Synthesis of oligonucleotide derivatives carried out using mechanical sintetizador nucleic acid (ABI model 392 DNA/RNA sintetizador - product firm Perkin-Elmer Corporation), in the amount of 1.0 µmol. Solvents, reagents and concentrations of fosfoamidaza in each cycle of the synthesis are the same as in the synthesis of natural oligonucleotides. Solvents, reagents and postholiday natural nucleosides are products of fitem repetition condensation compounds, obtained in example 9, or amidites containing 4 nucleotides, 5'-hydroxycytidine obtained by removal of the protective dimethoxymethylsilyl group at the 5'-O-4,4'-dimethoxypyrimidine (1.0 mmol) using trichloroacetic acid, and 3'-hydroxy-group thymidine was added to the media CGP.

The cycle of synthesis described below:

1) remove trailvoy groups in a mixture of trichloroacetic acid/dichloromethane, 35;

2) the combination of fosfoamidaza (approximately 20 EQ) in a mixture of tetrazole/acetonitrile, 25 or 10 min;

3) floor - in the mix 1-Mei/tetrahydrofuran, acetic anhydride/pyridine/tetrahydrofuran, 15;

4) oxidation in a mixture of iodine/water/pyridine/tetrahydrofuran, 15 S.

In the above cycle 2), when using the compound obtained in example 9, the reaction time is 10 min, and when they use postholiday, the reaction time is 25 C.

After synthesis of the desired sequence containing oligonucleotide derivatives, remove the 5'-dimethoxytrityl group and then the media containing the desired product, traditionally treated with concentrated aqueous ammonia solution in order to separate the oligomer from the media and delete zenatello group, cosine removed from the oligomer. Oligonucleotide derivative purified by HPLC with reversed phase (HPLC: LC-VP - product firms Shimazu Corp.; column: Wakopak WS-DNA - product firms Wako Pure Chemical Industry Ltd.), to get the desired oligonucleotide.

In accordance with this method of synthesis get the following oligonucleotide sequence (oligonucleotide which in the following is designated as "oligonucleotide 1") in an amount of 0.23 mmol, yield 23%: 5'-GCGTTTTTTGCT-3' (illustrative sequence number 2 in the sequence listing), in which the functional group of the sugar thymidine rooms grounds from 4 to 8 is a 2'-0,4'-C-ethylene.

Comparative example 1. 3,5-Di-O-benzyl-4-triftoratsetilatsetonom-1,2-O-isopropylidene-alpha-D-altropane-furanose

Anhydrous pyridine (of 0.60 ml, 7.5 mmol) is added to a solution of 3,5-di-O-benzyl-4-hydroxymethyl-1,2-O-isopropylidene-alpha-D-erythropoietins (2000 mg, 5.0 mmol) in anhydrous dichloromethane (50 ml) and added dropwise to the anhydride of triftoratsetata (1010 mg, 6.0 mmol) in a nitrogen atmosphere at -78C, the mixture is stirred for 40 minutes, the Reaction mixture was distributed between methylene chloride and saturated aqueous sodium hydrogen carbonate solution (approximately 100 ml). The organic layer is or sodium chloride (approximately 100 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo, obtaining a white powder (2520 mg, to 4.73 mmol, 95%), which is used in the next reaction without further purification.

An NMR spectrum1H (400 MHz, CDCl3): of 1.34 (3H, s), and 1.63 (3H, s), 3,48 (1H, d, 10 Hz), 3,53 (1H, d, 10Hz), is 4.21 (1H, d, 5.0 Hz), and 4.5 (4H, m), 4,74 (1H, d, 12 Hz), 4,80 (1H, d, 12 Hz), free 5.01 (1H, d, 12 Hz), 5,73 (1H, q, j 4.6 Hz), and 7.3 (10H, m).

Comparative example 2. 3,5-Di-O-benzyl-4-lanmeter-1,2-O-isopropylidene-alpha-D-eritropeniya

The compound obtained in comparative example 1 (2520 mg, to 4.73 mmol), dissolved in dimethyl sulfoxide (50 ml) at 90C. To this solution was added sodium cyanide (463 mg, 9,46 mmol) at room temperature and the mixture was stirred at 50C for 3 hours. The reaction mixture was partitioned between water (approximately 100 ml) and ethyl acetate (about 100 ml). The organic layer was washed with saturated aqueous sodium chloride (approximately 100 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate = 4:1) to obtain colorless oil (1590 mg, to 3.89 mmol, 82%).

An NMR spectrum1H (400 MHz, Dl3): OF 1.34 (3H,57 (1H, m), 4,58 (1H, d, 12 Hz), was 4.76 (1H, d, 12 Hz), 5,73 (1H, d, and 3.7 Hz), and 7.3 (10H, m).

Comparative example 3. 3,5-Di-O-benzyl-4-formylmethyl-1,2-O-isopropylidene-alpha-D-eritropeniya

Toluene solution (1.5 M) isobutyleneisoprene (2 ml, 3.0 mmol) is slowly added dropwise into a solution of the compound obtained in comparative example 2 (610 mg, 1,49 mmol), in dichloromethane (10 ml) in a nitrogen atmosphere at -78C, the mixture was stirred at -78C for 1 h and then warmed to room temperature. To the reaction mixture is added methanol (5 ml) and saturated aqueous ammonium chloride solution (about 20 ml) and the mixture stirred for 30 minutes, the Reaction mixture was extracted with ethyl acetate (approximately 30 ml). The organic layer was washed with saturated aqueous sodium bicarbonate (approximately 30 ml) and saturated aqueous sodium chloride (approximately 30 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo, receiving product, which is used in the next reaction without further purification.

Comparative Example 4. 3,5-Di-O-benzyl-4-hydroxyethyl-1,2-O-isopropylidene-alpha-D-eritropeniya

Borohydride sodium NaBH4(7,6 mg, 0.2 mmol what camping is stirred at room temperature for 1 hour. The reaction mixture is distributed between ethyl acetate (about 10 ml) and water (approximately 10 ml) and the organic layer was washed with saturated aqueous sodium chloride (approximately 10 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate = 2: 1) to obtain colorless oil (117 mg, 0,284 mmol, 75%).

An NMR spectrum1H (400 MHz, CDCl3): of 1.33 (3H, s) of 1.66 (3H, s) of 1.78 (1H, DDD, 4,0, 8,5, 15 Hz), of 2.51 (1H, DDD, 3,4, 6,4, 15 Hz) and 3.31 (1H, d, 10Hz), of 3.54 (1H, d, 10 Hz), 3,80 (2H, m), of 4.13 (1H, q, j 5.3 Hz), 4,43 (1H, d, 12 Hz) to 4.52 (1H, d, 12 Hz), 4,55 (1H, d, 12 Hz) and 4.65 (1H, DD, 4,0, a 5.3 Hz), of 4.77 (1H, d, 12 Hz), 5,77 (1H, d, 4.0 Hz), and 7.3 (10H, m).

FABMS (mNBA): 415 (M+N)+. []D+57,4(0,91, in methanol).

Comparative Example 5. 3,5-Di-O-benzyl-4-formyl-1,2-O-isopropylidene-alpha-D-eritropeniya

Oxalicacid (6,02 ml of 69.0 mmol) is added to methylene chloride (200 ml), cooled to -78C. To this solution is added dropwise a solution of dimethylsulfoxide (7,87 ml, 110 mmol) in anhydrous methylene chloride (100 ml). After stirring for 20 min the mixture was added dropwise a solution of 3,5-di-O-benzyl-1,2-O-isopropylidene-alpha-D-Tu, the reaction mixture was added triethylamine (28 ml, 200 mmol) and the mixture is slowly heated to room temperature. The reaction mixture was partitioned between dichloromethane and water (about 300 ml). The organic layer was washed with water (about 300 ml) and saturated aqueous sodium chloride (approximately 300 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate = 5:1) to obtain colorless oil (8310 mg, to 20.88 mmol, 91%).

An NMR spectrum1H (400 MHz, Dl3): of 1.35 (3H, s) to 1.60 (3H, s), 3,61 (1H, d, 11 Hz), 3,68 (1H, d, 11 Hz), 4,37 (1H, d, 4.4 Hz), 4,46 (1H, d, 12 Hz) to 4.52 (1H, d, 12 Hz), 4,59 (1H, d, 12 Hz), 4,59 (1H, DD, 3,4, 4,4 Hz), 4,71 (1H, d, 12 Hz), of 5.84 (1H, d, 3.4 Hz), and 7.3 (10H, m), to 9.91 (1H, s).

FABMS (mNBA): 397 (M-N)+, 421(M+Na)+.

[]D+27,4° (of 0.51, methanol).

Comparative Example 6. 3,5-Di-O-benzyl-4-vinyl-1,2-O-isopropylidene-alpha-D-eritropeniya

Toluene 0.5 M solution of the reagent Tebbe (44 ml, 22 mmol) are added to a solution of the compound obtained in comparative example 5 (8310 mg, to 20.88 mmol) in anhydrous tetrahydrofuran (300 ml) under nitrogen atmosphere at 0C and the mixture was stirred at 0C for 1 hour. To reactionaries filtered through celite, to remove the precipitate, and the precipitate washed with diethyl ether (about 100 ml). The organic layer is dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatography on basic alumina using dichloromethane to obtain the crude product, which is optionally purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate =8:1-5:1) to obtain colorless oil (5600 mg, 14,14 mmol, 68%).

An NMR spectrum1H (400 MHz, CDCl3): of 1.28 (3H, s), of 1.52 (3H, s), and 3.31 (1H, d, 11 Hz), 3,34 (1H, d, 11 Hz), 4,25 (1H, q, j 4.9 Hz), and 4.40 (1H, d, 12 Hz) to 4.52 (1H, d, 12 Hz), of 4.57 (1H, DD, of 3.9, and 4.9 Hz), 4,59 (1H, d, 12 Hz), was 4.76 (1H, d, 12 Hz), the 5.25 (1H, DD, 1,8, 11 Hz), 5,52 (1H, DD, 1,8, 18 Hz), USD 5.76 (1H, q, j 3.9 Hz), of 6.20 (1H, DD, 11, 18 Hz), and 7.3 (10H, m).

FABMS (mNBA): 419(M+Na)+.

Comparative Example 7. 3,5-Di-O-benzyl-4-hydroxyethyl-1,2-O-isopropylidene-alpha-D-eritropeniya

Tertrahydrofuran ring 0.5 M solution of 9-BBN (9-borabicyclo[3,3,1]nonan) (80 ml, 40 mmol) is added dropwise to a solution of the compound obtained in comparative example 6 (5500 mg, 13,89 mmol) in anhydrous tetrahydrofuran (200 ml) under nitrogen atmosphere and the mixture is stirred at room temperature overnight. To the reaction mixture, water is added until there is no further allocation of ha is hydrogen, maintaining the temperature between 30 and 50C. This mixture is stirred for 30 min and partitioned between saturated aqueous sodium chloride (approximately 200 ml) and ethyl acetate (200 ml). The organic layer was washed with a neutral buffer solution of phosphoric acid (approximately 200 ml) and saturated aqueous sodium chloride (approximately 200 ml) and dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate = 2:1-1:1) to obtain colorless oil (5370 mg, 12,97 mmol, 93%).

An NMR spectrum1H (400 MHz, CDCl3): of 1.33 (3H, s) of 1.66 (3H, s) of 1.78 (1H, DDD, 4,0, 8,5, 15 Hz), of 2.51 (1H, DDD, 3,4, 6,4, 15 Hz) and 3.31 (1H, d, 10Hz), of 3.54 (1H, d, 10 Hz), 3,80 (2H, m), of 4.13 (1H, q, j 5.3 Hz), 4,43 (1H, d, 12 Hz) to 4.52 (1H, d, 12 Hz), 4,55 (1H, d, 12 Hz) and 4.65 (1H, DD, 4,0, a 5.3 Hz), of 4.77 (1H, d, 12 Hz), 5,77 (1H, d, 4.0 Hz), and 7.3 (10H, m).

FABMS (mNBA): 415(M+N)+. []D+57,4(0,91, in methanol).

Comparative Example 8. 3,5-Di-O-benzyl-4-(para-toluensulfonate)-1,2-O-isopropylidene-alpha-D-eritropeniya

The triethylamine (1.8 ml, 13 mmol), dimethylaminopyridine (30 mg, 0.25 mmol) and para-toluensulfonate (858 mg, 4.5 mmol) are added to lwala (1035 mg, 2.5 mmol), in anhydrous dichloromethane (35 ml) under nitrogen atmosphere at 0C and the mixture is stirred at room temperature overnight. The reaction mixture was partitioned between dichloromethane and saturated aqueous sodium hydrogen carbonate solution (approximately 100 ml). The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution (approximately 100 ml) and saturated aqueous sodium chloride (approximately 100 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate = 3:1) to obtain colorless oil (1340 mg, 2.6 mmol, 94%).

An NMR spectrum1H (400 MHz, Dl3): of 1.33 (3H, s), for 1.49 (3H, s), 1,99 (1H, dt, 7.6 and 15 Hz), 2,47 (3H, s), 2,60 (1H, DDD, 5,7, 7,6, 15 Hz), or 3.28 (1H, d, 10Hz), of 3.45 (1H, d, 10 Hz), 4,11 (1H, d, 5.3 Hz), 4,32 (2H, m), 4,42 (1H, d, 12 Hz), 4,50 (1H, d, 12 Hz), of 4.54 (1H, d, 12 Hz), to 4.62 (1H, DD, 4,0, 5,2 Hz), was 4.76 (1H, d, 12 Hz), 5,74 (1H, d, 4.0 Hz), 7,3 (N, m), 7,78 (2H, d, 8,3 Hz).

FAB-MAS(mNBA): 569(M+N)+.

Comparative Example 9. 1,2-Di-O-acetyl-Z,5-di-O-benzyl-4-(para-toluensulfonate)-alpha-D-eritropeniya

Acetic anhydride (1,88 ml, 20 mmol) and concentrated sulfuric acid (0.01 ml) are added to a solution of the compound, the Oh temperature for 1 hour. The reaction mixture was poured into water (60 ml) in an ice bath, and stirred for 30 min, then partitioned between saturated aqueous sodium chloride (approximately 100 ml) and ethyl acetate (about 100 ml). The organic layer was washed with a neutral buffer solution of phosphoric acid, saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and then concentrated. The residue is purified chromatographically on silica gel (using a mixture of hexane : ethyl acetate = 2:1) to obtain colorless oil (1290 mg, 2,11 mmol, 89%,:=1:5).

An NMR spectrum1H (400 MHz, Dl3): (-derivative) to 1.86 (3H, s), is 2.05 (3H, s), of 2.08 (1H, m) to 2.18 (1H, m), 2,42 (3H, s), 3,30 (1H, d, 10 Hz) to 3.33 (1H, d, 10 Hz) to 4.23 (1H, q, j 5.1 Hz), 4,24 (2H, m), 4,42 (2H, s), of 4.45 (1H, d, 12 Hz), 4,55 (1H, d, 12 Hz), 5,28 (1H, d, 5.1 Hz), 6,01 (1H, s), and 7.3 (N, m), 7,73 (2H, d, 8,3 Hz).

FAB-MAS(mNBA): 613(M+N)+.

Comparative Example 10. 2'-O-Acetyl-3',5'-di-O-benzyl-4'-para-toluensulfonate-5-methyluridine

Trimethylsilylethynyl thymine (500 mg, about 2 mmol), obtained in accordance with the method of H. Vorbrggen, K. Krolikiewicz and Century Bennua (Chem. Ber., so 114, S. 12,2-dichloroethane (15 ml) at room temperature under nitrogen atmosphere. Trimethylsilyltrifluoromethane (0,36 ml, 2 mmol) is added dropwise to the mixture and the mixture is stirred at a temperature of 50C for 1 hour. To the reaction mixture is added saturated aqueous sodium hydrogen carbonate solution (approximately 50 ml) and the mixture filtered through celite. Add to the filtrate dichloromethane (approximately 50 ml). The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution (approximately 50 ml) and saturated aqueous sodium chloride (about 50 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel using hexane :ethyl acetate = 1,2: 1) to obtain a colorless amorphous solid (432 mg, 0.64 mmol, 60%).

An NMR spectrum1H (400 MHz, CDCl3): of 1.52 (3H, d, and 0.9 Hz), was 1.94 (1H, dt, 7.5 and 15 Hz) to 2.06 (3H, s), of 2.23 (1H, dt, 6.0 and 15 Hz), 2,42 (3H, s) to 3.38 (1H, d, 10 Hz) to 3.67 (1H, d, 10 Hz), 4,17 (2H, m), 4,36 (1H, q, j 6.0 Hz), to 4.41 (1H, d, 12 Hz), of 4.44 (1H, d, 12 Hz), 4,48 (1H, d, 12 Hz), 4,58 (1H, d, 12 Hz), of 5.39 (1H, DD, of 5.1 and 6.0 Hz), 6,04 (1H, d, 5.1 Hz), and 7.3 (N, m), 7,73 (2H, dt, of 1.8 and 8.3 Hz), 8,18 (1H, s).

FAB-MAS (mNBA): 679(M+H)+.

Comparative Example 11. 2'-O-Acetyl-3',5'-di-O-benzyl-4'-para-toluensulfonate-4-N-benzoylation

Trimethylsilylpropyne the iewicz and B. Bennua (Chem. Ber., so 114, S. 1234-1255 (1981)), are added to a solution of the compound obtained in comparative example 9 (383 mg, 0,626 mmol) in anhydrous 1,2-dichloroethane (4 ml). To this mixture trimethylsilyltrifluoromethane (of 0.18 ml, 0,995 mmol) at 0C and the mixture was stirred at 50C for 1 hour. To the mixture is added saturated aqueous sodium hydrogen carbonate solution (approximately 10 ml) and methylene chloride (approximately 20 ml) and the mixture is then stirred. The resulting white precipitate was filtered through celite. The organic layer of the filtrate was washed with saturated aqueous sodium chloride (approximately 20 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo to obtain a colorless amorphous solid (397 mg, 83%).

An NMR spectrum1H (400 MHz, CDCl3): to 8.70 (1H, br), 8,18 (1H, q, j 7.4 Hz), 7,87 (2H, q, j 7.5 Hz), 7,72 (2H, d, 8,3 Hz), to 7.61-EUR 7.57 (1H, m), 7,51-of 7.48 (2H, m), 7,43-7,21 (13H, m), of 6.02 (1H, d, 2,9 Hz), of 5.40 (1H, DD, 5,8, 2,9 Hz), of 4.57 (1H, d, 11 Hz), 4,39 (1H, d, 11 Hz), 4,32-to 4.28 (3H, m), 4,19-4,16 (2H, m), of 3.69 (1H, d,11 Hz), and 3.31 (1H, d, 11 Hz), is 2.40 (3H, s), 2,30-of 2.23 (1H, m) to 2.06 (3H, s), 1,95-1,89 (1H, m).

FAB-MAS (mNBA): 768(M+H)+.

Comparative Example 12. 2'-O-Acetyl-3',5'-di-O-benzyl-4'-para-toluensulfonate-6-N-benzyladenine

Trimethylsilyl, K. Krolikiewicz and Century Bennua (Chem. Ber., so 114, S. 1234-1255 (1981)), are added to a solution of the compound obtained in comparative example 9 (600 mg, 0.98 mmol) in anhydrous 1,2-dichloroethane (15 ml) at room temperature under nitrogen atmosphere. After adding dropwise trimethylsilyltrifluoromethane (0,36 ml, 2 mmol) to the mixture, the mixture is stirred at 50°C for 4 hours. To the reaction mixture is added saturated aqueous sodium hydrogen carbonate solution (approximately 50 ml) and dichloromethane (50 ml) and the mixture is distributed between the two layers. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution (approximately 50 ml) and saturated aqueous sodium chloride (about 50 ml), dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of dichloromethane : methanol = 50:1) to obtain a colorless amorphous solid (405 mg, 0.51 mmol, 52%).

An NMR spectrum1H (400 MHz, CDCl3): 2,0 (1H, m) to 2.06 (3H, s), 2,32 (1H, dt, 6.0 and 15 Hz), is 2.40 (3H, s) to 3.36 (1H, d, 10 Hz) to 3.58 (1H, d, 10 Hz), 4,22 (2H, m), 4,39 (1H, d, 12 Hz), of 4.45 (1H, d, 12 Hz), 4,47 (1H, d, 12 Hz), 4,59 (1H, d, 12 Hz), to 4.62 (1H, q, j 5.6 Hz), 5,94 (1H, DD, 4.5 and 5.6 Hz), 6,21 (1H, d, 4.5 Hz), 7,2-7,3 (N, m), 7,54 (2H, m), a 7.62 (1H, dt, 1,2 and 6.2 Hz), 7,7-O-Acetyl-3',5'-di-O-benzyl-4'-para-toluensulfonate

Trimethylsilylethynyl uracil (200 mg, about 0.8 mmol), which was obtained according to method H. Vorbrggen, K. Krolikiewicz and Century Bennua (Chem. Ber., so 114, S. 1234-1255 (1981)), are added to a solution of the compound obtained in comparative example 9 (200 mg, 0,327 mmol) in anhydrous 1,2-dichloroethane (8 ml) at room temperature under nitrogen atmosphere. After adding to the mixture dropwise trimethylsilyltrifluoromethane (0,145 ml, 0.8 mmol), the mixture is stirred at a temperature of 70C for 1 hour. To the reaction mixture is added saturated aqueous sodium hydrogen carbonate solution (approximately 10 ml), the mixture is filtered through celite and to the filtrate add dichloromethane (about 10 ml). The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and then concentrated in vacuo. The residue is purified chromatographically on silica gel (using a mixture of dichloromethane : methanol = 100:2) to obtain a colorless oil (199 mg, 0,299 mmol, 92%).

An NMR spectrum1H (400 MHz, CDCl3): 1,94 (1H,dt, 7.4 and 15 Hz), 2,07 (3H, s), of 2.23 (1H, dt, 5.9 and 15 Hz), 2,43 (3H, s) to 3.36 (1H, q, j 10 Hz), the 3.65 (1H, q, j 10 Hz), 4,17 (2H, DD, 6, and 7 Hz), or 4.31 (1H, d, 5,9 Hz), to 4.38 (1H, d, 161 (1H, d, 8.2 Hz), 7,74 (1H, d, 8,3 Hz), 8,14 (1H, Shir. C).

FAB-MAS(mNBA): 665(M+N)+.

Comparative Example 14. 2'-O-Acetyl-3',5'-di-O-benzyl-4'-para-toluensulfonate-4-N-benzoyl-5-methylcytidine

Trimethylsilylethynyl benzoyl - 5-methylcytosine (400 mg, approximately 1.2 mmol), which was obtained according to method H. Vorbrggen, K. Krolikiewicz and Century Bennua (Chem. Ber., so 114, S. 1234-1255 (1981)), are added to a solution of the compound obtained in comparative example 9 (400 mg, 0,653 mmol) in anhydrous 1,2-dichloroethane (6 ml). After adding to the mixture trimethylsilyltrifluoromethane (0,180 μl, 1.0 mmol) at 0The mixture is stirred at a temperature of 50C for 1 hour. The reaction mixture is heated to room temperature. To the reaction mixture is added saturated aqueous sodium hydrogen carbonate solution (approximately 5 ml) and methylene chloride (approximately 10 ml) and the mixture is stirred. The mixture is filtered through celite to remove the white precipitate. The organic layer of the filtrate was washed with saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and then concentrated in vacuo, obtaining a colorless amorphous solid (320 mg, 0,409 mmol, 63%).

An NMR spectrum1H (400 MHz, CDCl3

FAB-MAS (mNBA): 782 (M+N)+.

Comparative Example 15. 2'-O-Acetyl-3',5'-di-O-benzyl-4'-para-toluensulfonate-2-N-isobutylamine

Trimethylsilylethynyl isobutylbenzene (650 mg, about 1.5 mmol), which was obtained according to method H. Vorbrggen, K. Krolikiewicz and Century Bennua (Chem. Ber., so 114, S. 1234-1255 (1981)), are added to a solution of the compound obtained in comparative example 9 (400 mg, of 0.65 mmol) in anhydrous 1,2-dichloroethane (10 ml) at room temperature under nitrogen atmosphere. After adding to the mixture trimethylsilyltrifluoromethane (0.2 ml, 1.2 mmol), the mixture was stirred at 50C for 4 hours. To the reaction mixture is added saturated aqueous sodium hydrogen carbonate solution (approximately 5 ml), the organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and then concentrated in vacuo, receiving product, which is used in the next reaction without further purification.

Test example 1. Determination of the melting temperature TM

The solution is (1) and 4 μm complementary DNA (hereinafter oligonucleotide (2)) with the sequence of the complementary chain (sequence: 5'-AGCAAAAAACGC-3' (sequence No. 1 in the sequence listing), or complementary RNA (hereinafter oligonucleotide (3) ) with the sequence 5'-AGCAAAAAACGC-3' (sequence No. 1 in the sequence listing) is heated in a boiling water bath and cooled slowly to room temperature for approximately two hours. Then the solution is heated and determine the absorbance at 260 nm using a spectrophotometer (UV-3100PC, product of Shimadzu Corp.). The sample is heated cuvette (cuvette thickness 1.0 cm, jacketed cylindrical type) circulating water heated in a thermostat (Haake FE2 - product firm ECO Corp.), to determine the growth temperature using a digital thermometer (firm S SK1250MC). The temperature was raised from 20 to 95With and for each increase in temperature of 1To measure the absorption at 260 nm, which corresponds to the maximum absorption. As a control using the DNA of natural origin (hereinafter oligonucleotide (4)), with the sequence 5'-GCGTTTTTTGCT-3' (sequence No. 2 in the sequence listing), which represents the same sequence, which oligonucleotide (1) (compound of example 29).

The temperature at which the intensity changes by 1°C reaches its maximum, is taken by the melting temperature TM, and sposobnost.

Below shows the results of measurement of the melting temperature TM of the oligonucleotide (4) (DNA of natural origin) and the oligonucleotide (1) (compound of example 29) with respect to the oligonucleotide (2) (complementary DNA) and the oligonucleotide (3) (complementary RNA).

As can be seen from the above table, for oligonucleotide analogue of the present invention are observed significantly higher values of the melting temperature TM, as well as a significantly higher ability to form complex with the complementary chain compared with the DNA of natural origin.

Test example 2. Determination of resistance to nucleases

Oligonucleotide are mixed in a buffer solution with exonuclease or endonuclease and incubated at 37With over a certain period of time. To stop reaction, add ethylenediaminetetraacetic acid (EDTA) and heated at 100C for 2 minutes, the Number of oligonucleotide remaining in the mixture was determined by HPLC on a column of reversed phase and to determine the changes in the number of the oligonucleotide in time, in the presence of nucleases.

Oligonucleotide analogues of the present from the STI to nucleases.

(a) synthesis of the reference compound is 2'-O,4-C-methylene nucleoside of formula (1)

Oligonucleotide containing 2'-O,4-C-methylenchlorid formula (1) was synthesized according to the method, opened in W98/39352 and its resistance to nucleases compared with the stability of the oligonucleotide containing nucleosides of the present invention.

Oligonucleotide A: 5'-tlt ttt ttt txt-31

b) Synthesis of compounds of the present invention

Oligonucleotide, containing nucleosides of the invention, synthesized according to the method, opened in the description of this application.

Oligonucleotide: 5'-ttt ttt ttt tyt-3'

(C) Methods of analysis for resistance to nuclease and the results obtained. The stability of the oligonucleotide and the oligonucleotide In to action of snake venom phosphodiesterase was determined by the method described in Comparative example 2. Oligonucleotide in the amount of 26 μg/ml was added in a solution containing 50 mm Tris-HCL (pH 8.0) and 10 mm MgCl2. Resistance to nucleases was determined as the percentage ratio of the number of remaining oligonucleotides to their original amounts. The results obtained are presented in Table 4.

From this we can conclude that the oligonucleotide In the present invention has a significantly higher resistance to nucleases than the known oligonucleotide A. Such a high resistance to nucleases of the oligonucleotide according to the invention is not obvious to a person skilled in the art. As the role of ring structures of nucleosides of the invention is not purely structural factor, apparently, this is the hallmark of the invention in comparison with the prior art.

Test example 4. Determining the ability of the oligonucleotides of the formula (I) to the formation of a duplex with a complementary chain and resistance to nucleases

The oligonucleotide of the present invention has the formula (I)

where a represents (CH2)2group.

(a) Synthesis of known compounds

References cited in the International search report, for example in Tetrahedron, 1998, 54, 3607, etc. are disclosed 2'-O,4'-C-methylenchlorid formula (II)

Oligonucleotide containing 2'-O,4'-C-methylenchlorid formula (II) was synthesized according to the method, a revelation is rasih nucleosides of the invention.

Oligonucleotide A: 5'-ttt ttt ttt txt-3'

(b) Synthesis of compounds of the present invention

Oligonucleotide and oligonucleotide With containing nucleoside according to the invention synthesized in accordance with the following scheme, as well as the methods indicated in the description of this application.

Oligonucleotide: 5'-ttt ttt ttt tyt-3'

Oligonucleotide: 5'-gcg ttt tty gct-3'

The schema. Synthesis of 2'-O,4'-C-propylenimine-3'-O-phosphoramidites and oligonucleotides. Reagents and conditions: (i) (COCl)2, DMSO, Et3N, CH2CL2, 78C, 99%; (ii) (EtO)2P(O)CH2COOEt, NaH, THF, 86%; (iii) Pd(OH)2; H2(3.5 lb/in2), Meon, 71%; (iv) L-selectride, THF, 85%; (v) TsCl, Et3N, CH2Cl2, 77%; (vi) Ac2O, H2SO4, AcOH, 79% (mainly-anomer); (vii) sililirovany thymine, TMSOTf, ClC2H4Cl, delegacia, 88%; (viii) 1M NaOH, pyridine-H2O; (ix) TBAF; THF, 82% (stage 2); x) H2Pd(OH)2, MeOH, 59%; (xi) DMTrCl, pyridine, CH2Cl2, quantitative xii) ((iPr)2N)2P(OC2H4CN), tetrazole N,N-Diisopropylamine, quantitative; xiii) synthesizer DNA/RNA.

5-O-(tert-Butyldiphenylsilyl)-4-formyl-3-O2Cl2(200 ml) at -78C. To the resulting solution was bury a solution of DMSO (11,4 ml, 160 mmol) in anhydrous CH2Cl2(100 ml). After stirring for 20 minutes, to the resulting mixture was bury a solution of compound 1 (Obica,S; Morio, K; Hari, Y.; Imanishi, T. Chem. Commun., 1999, 2423)(26,3 g of 47.9 mmol) in anhydrous CH2CL2(100 ml) and the resulting mixture was stirred for 30 minutes. In the reaction mixture were added triethylamine (42 ml, 300 mmol) and the resulting mixture was slowly heated to room temperature. The reaction mixture was cooled H2O. the Organic layer was washed with water and brine, dried over MgSO4and then evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (hexane : Eoas=7:1) with the formation of compound 2 as a colorless oil (26,1 g of 47.8 mmol, 99%).

Range13C-NMR (125 MHz, CDCl3):19,21, 26,13, 26,66, 26,76, 63,05, 72,73, 78,57, 79,08, 90,62, 104,91, 114,15, 127,76, 127,79, 127,83, 128,07, 128,51, 129,81, 129,86, 132,51, 132,82, 135,49, 135,57, 136,99, 100,17.

1H NMR (400 MHz, CDCl3):0,98 (N, C) to 1.37 (3H, s) of 1.62 (3H, s), with 3.79 (1H, d, 11 Hz), 3,88 (1H, d, 11 Hz), of 4.54 (1H, d, 4.4 Hz), to 4.62 (1H, d, 12 Hz) and 4.65(1H, DD, and 3.7 and 4.4 Hz), 4,74 (1H, d, 12 Hz), by 5.87 (1 H, d, and 3.7 Hz), and 7.3 and 7.6 (15 NM), of 9.89(1H, s).

IR(film)+
569,2335 found 569,2354.

5-O-(tert-Butyldiphenylsilyl)-4-(2-etoxycarbonyl-(E)-vinyl)-3-O-benzyl-1,2-O-isopropylidene-a-D-eritropeniya (3). Sodium hydride (60% in mineral oil, 2.3 g, about 57,4 mmole) was added to ethyl ether diethylphosphonate acid (11.5 ml, 57,4 mmole) in anhydrous THF (200 ml) at 0C. After 10 minutes of stirring the solution was bury a solution of compound 2 (26,1 g, 47,8 mmole) in anhydrous THF (200 ml) and the resulting mixture was stirred for 30 minutes at room temperature. The reaction mixture was cooled with water and was extracted with t. The organic layer was washed with brine, dried over MgSO4and then evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (hexane : tO=7:1) with the formation of 3 as a colorless oil (25,4, 41,2 mmole, 86%).

Range13C-NMR (125 MHz, Dl3):14,23, 19,25, 25,66, 26,07, 26,77, 60,21, 65,60, 72,53, 77,21, 78,08, 86,68, 103,99, 113,48, 122,66, 127,68, 127,73, 127,80, 127,96, 128,49, 129,73, 129,71, 133,12, 135,49, 135,56, 145,51, 166,21.

Range1H-NMR (400 MHz, CDCl3):1,04 (N, C) of 1.30 (3H, t, 7,3 Hz) of 1.45 (3H, s) of 3.54 (1H, d,10 Hz) to 3.58 (1H, d,10 Hz) and 4.2 (2H, m), 4,39 (1 H, d, 6.3 Hz), 4,59 (1H, d, 10 Hz), 4,58 (1H, DD, 4.4 and 6.3 Hz), of 4.77(1H, d, 10 Hz), 5,90 (1H, d, a 4.4 Hz), 6,24 (1H, d, 16 Hz), and 7.3 to 7.7(15 NM,>/p>ESI-HRMS (positive): calculated for C36H44O7SiNa [M+Na]+639,2754 found 639,2755.

5-O-(tert-Butyldiphenylsilyl)-4-(2-ethoxycarbonylethyl)-3-O-benzyl-1,2-O-isopropylidene-a-D-eritropeniya (4). To a solution of compound 3 (26,0 g, 42,14 mmole) in tOc (100 ml) was added 20% palladium hydroxide on coal (8.0 g). The mixture was stirred in hydrogen atmosphere at a pressure of 3.5 lb/in2within 10 hours. The reaction mixture was filtered and the filtrate evaporated in vacuum. The residue was purified by chromatography on silica gel (hexane : tO=7:1) with the formation of compound 4 as a colorless oil (18.5 g, and 29.9 mmole, 71%) and dibenzylamine compound (6.0 g, 11,34 mmole, 27%).

13C-NMR (125 MHz, Dl3):14,16, 19,20, 26,19, 26,59, 26,80, 27,00, 29,02, 60,12, 66,29, 72,35, 77,97, 79,16, 86,88, 104,17, 113,27, 127,73, 127,76, 128,40, 129,68, 129,76, 132,89, 133,17, 135,53, 135,61, 137,92, 173,88.

Range1H-NMR (400 MHz, CDCl3): 0,98 (N, C) of 1.18 (3H, t, 7.0 Hz), of 1.35 (3H, s) of 1.64 (3H, s), equal to 1.82 (1H, m) of 2.16 (1H, m), 2,52 (2H, m), 3,40 (1H, d, 11 Hz), 3,61 (1H, d, 11 Hz), Android 4.04 (1H, d, 12 Hz), 5,78 (1H, d, 4.4 Hz), and 7.3 to 7.7 (15 N, m).

IR(KBR)max: 2933, 2858, 1733, 1111, 1025, 703 cm-1.

FAB-MS (mNBA): 617[M-H], 641[M+Na]+.

ESI-HRMS (positive): calculated for C36H46O7SiNa [M+Na]+3and brine, dried over MgSO4and evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (hexane : tOc=4:1) to obtain compound 5 as colorless oil (1.55 g, 2,69 mmole, 85%).

IR(film)max: 3487, 2934, 2858, 1112, 1025, 703-1.

Range13C NMR (125 MHz, CDCl3): 19,21, 26,06, 26,58, 26,77, 26,80, 27,51, 62,57, 65,97, 72,42, 77,83, 79,15, 87,95, 104,13, 113,14, 127,68, 127,73, 127,82, 128,42, 129,70, 129,75, 132,92, 133,29, 135,53, 135,65, 137,95.

Range1H-NMR (400 MHz, CDCl3): 0,98 (N, C) of 1.35 (3H, s), and 1.63 (3H, s), 1,54 is 1.70 (2H, m) of 1.97 (1H, t, 6,1 Hz), 2,17 (1H, m), 3,44 (1H, d, 11 Hz), of 3.56 (2H, m), 3,74 (1H, 11 Hz), 4,35 (1H, q, j 1 Hz), 4,59 (1H, d, 12 Hz), and 4.68 (1 H, DD, of 4.4 and 5.1 Hz), 4,80 (1H, d, 12 Hz), 5,79 (1H, d, 4.4 Hz), and 7.3 to 7.7(15 NM, m).

FAB-HRMS (mNBA): calculated for C34H44O6SiN [M+Na]+599,2805 found 599,2819.

5-O-(tert-Butyldiphenylsilyl,0 mmole) and chloride p-toluensulfonyl (286 mg, 1.5 mmole) was added to a solution of compound 5 (610 mg, of 1.06 mmole) in anhydrous CH2CL2(10 ml) under nitrogen atmosphere at 0C and the resulting mixture was stirred over night at room temperature. The reaction mixture was distributed between CH2CL2and saturated NaHCO3. The organic layer was washed with brine, dried over MgSO4and then evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (hexane : tO=7:1) with the formation of 6 as a colorless oil (600 mg, about 0.82 mmole, 77%).

IR(film)max: 2933, 2858, 1360, 1176, 1112, 704cm-1. Range13C-NMR (400 MHz, CDCl3)19,17, 21,56, 23,20, 26,17, 26,73, 26,77, 27,80, 66,33, 71,09, 72,37, 78,04, 79,25, 87,20, 104,16, 113,13, 127,70, 127,76, 127,81, 127,87, 128,42, 129,70, 129,79, 132,85, 133,11, 133,13, 135,51, 135,58, 137,89, 144,44.

Range1H-NMR (400 MHz, CDCl3)as 0.96 (N, C) of 1.32 (3H, s), 1.50 in (3), 1,42-of 1.56 (2H, m), 1,82 (N, m) is 2.05 (1H, m), 2,39 (3H, s) to 3.34 (1H, d, 11 Hz), of 3.56 (1H, d, 11 Hz), 3,90-4,00 (2H, m), 4,25 (1H, d, 5.1 Hz), of 4.54 (1H, d, 12 Hz), 4,63(1 H, DD, and 3.7 and 5.1 Hz), of 4.77 (1H, d, 12 Hz), 5,74 (1H, d, and 3.7 Hz), 7.3 to 7.8 for (N, m).

ESI-HRMS: calculated for C41H50O8SiNa [M+Na]+753,2893 found 753,2878.

5-O-(tert-Butyldiphenylsilyl)-4-(p-toluensulfonyl)-3-O-benzyl-1,2-di-O-0.01 ml) was added to a solution of compound 6 (600 mg, 0.82 mmol) in acetic acid (5 ml) and the resulting mixture was stirred at room temperature for 30 minutes. The reaction mixture was poured into H2About (10 ml) in a bath with ice and stirred for 30 minutes, then was extracted with EtOAc. The organic layer is washed with neutral phosphate buffer, a saturated solution of NaHCO3and brine, dried over MgSO4and then evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (hexane : tO=5:1) with the formation of predominantly-anomer of compound 7 as a colorless oil (500 mg, 0.65 mmole, 79%).

IR(film)max: 1748, 1362, 1220, 1176, 1112, 704 cm-1.

Range13C-NMR (125 MHz, CDCl3): 19,32, 20,75, 20,96, 21,58, 22,85, 26,90, 28,23, 67,30, 71,15, 73,50, 74,75, 79,07, 87,79, 97,70, 127,51, 127,77, 127,84, 128,43, 129,74, 129,82, 129,94, 132,71, 133,05, 133,24, 135,49, 135,59, 137,59, 144,50, 169,20, 169,71.

Range1H-NMR (400 MHz, CDCl3): 1,03 (N, s), 1.60-to 1,90 (4H, m), of 1.80 (3H, s) to 2.06 (3H, s), is 2.40 (3H, s), 3,53 (2H, s), 3,98 (2H, m), 4,34 (1H, d, 5.2 Hz), 4,50 (1H, d, 11 Hz), of 4.57 (1H, d, 11 Hz), 5,33 (1H, d, 5.2 Hz), between 6.08 (1H, s), 7,307,8 (N, m).

ESI-HRMS: calculated for C42H50O10SiNa [M+Na]+797,2792 found 797,2798.

5-O-(tert-Butyldiphenylsilyl)-4'-(p-toluensulfonyl the first method Vorbruggen (Vorbruggen,H.; Bennua, C. Chem. Ber., 1981, 114, 1279) was added to a solution of compound 7 (490 mg, of 0.64 mmole) in anhydrous 1,2-dichloroethane (15 ml) at room temperature under nitrogen atmosphere. TMSOTf (0.2 ml, 1.1 mmole) was bury in the mixture and the mixture was stirred at 50C for 3 hours. The reaction was stopped by adding a saturated solution of NaHCO3and filtered through celite. The organic layer was washed with saturated solution of NaHCO3and brine, dried over MgSO4and then evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (CH2Cl2:MeOH=100:3) to give 8 as a colorless amorphous solid (475 mg, of 0.56 mmole, 88%).

IR(KBR)max: 2931, 1748, 1693, 1361, 1228, 1176, 1111, 703 cm-1.

1H-NMR (400 MHz, CDCl3): 1,10 (N, C), 1,40-of 1.52 (2H, m) of 1.55 (3H, s), 1,72-to 1.82 (2H, m), of 2.08 (3H, s), is 2.40 (3H, s), 3,49 (1H, d, 12 Hz in), 3.75 (1H, d, 12 Hz), 3,92 (2H, m), 4,37 (1H, q, j 6.6 Hz), was 4.02 (1H, d, 12 Hz), of 4.57 (1H, d, 12 Hz), are 5.36 (1H, t, 6.6 Hz), 6,12 (1H, q, j 6.6 Hz), of 7.3 to 7.7 (N, m), 7,71 (1H, d, 2.2 Hz), 8,08 (1H, s).

ESI-HRMS: calculated for C45H52N2O10SiNa [M+Na]+863,3009 found 863, 2991.

3'-O-Benzyl-2'-O,4'-propylene-5-methyluridine (9). 3 ml of 1.0 N NaOH solution was added to a solution of 8 (475 mg, of 0.56 mmole) in pyridine (5 ml the Reaction mixture was neutralized by addition of aqueous 20% solution of acetic acid and was extracted with CH2Cl2. The organic layer is washed with neutral phosphate buffer and brine, and then evaporated in vacuum. The resulting crude compound was diluted in THF (5 ml) and to the solution was added 1.0 M solution of tetrabutylammonium fluoride in THF. The resulting reaction mixture was stirred for 6 hours at room temperature and then distributed between H2Oh and CH2CL2. The organic layer was washed with brine, dried over MgSO4and evaporated in vacuum. The residue was subjected to chromatographic purification on silicagel (CH2Cl2:MeOH=40:1) to give 9 as a colorless amorphous solid (180 mg, and 0.46 mmole, 82%).

Range1H-NMR (400 MHz, CDCl3): 1,50-1,90 (4H, m) to 1.87 (3H, s), 3,70 (1H, d, 12 Hz), 3,80 (1H, d, 12 Hz), of 4.05 (1H, m), 4,30 (1H, dt, 1.5 and 11 Hz), 4,37 (1H, d, 5,9 Hz), 4,42 (1H, d, 5,9 Hz), 4,56 (1H, d, 11 Hz), 4,82 (1H, d, 11 Hz), of 6.02 (1H, C), 7.3 to 7.4 (5H, m), 7,72 (1H, d, 1.5 Hz), of 8.25 (1H, broad).

IR(KBR)max: 3393, 2956, 1468, 1118 cm-1.

ESI-HRMS9 (positive): calculated for C20H25N2O6[M+H]+389,1713 found 389,1714.

FAB-MS(mNBA): 389[M+H]+.

2'-O,4'-Propylene-5-methyluridine (10). To a solution of compound 9 (175 mg, 0.45 mmole) in Meon (5 ml) was added 20% palladium hydroxide on the camping was filtered and the filtrate evaporated in vacuum. The residue was subjected to chromatographic purification on silica gel (CH2Cl2:MeOH=10:1) to give 10 as a colorless amorphous solid (81 mg, of 0.27 mmole, 59%).

1H-NMR (400 MHz, CD3OD): 1,58 (1H, m), 1,71 (1H, m) of 1.84 (2H, m), of 1.85 (3H, s) to 3.36 (1H, s), 3,66 (1H, d, 12 Hz), 3,74 (1H, d, 12 Hz), of 3.94 (1H, dt, 3,7 and 11 Hz), 4,11 (1H, d, 5,9 Hz), 4,28 (1H, m), 4,50 (1H, 5,9 Hz), 5,95 (1H, s), of 8.28 (1H, s).

FAB-MS (mNBA): 299[M+H]+.

ESI-HRMS (positive): calculated for C13H19N2About6[M+H]+299,1243 found 299,1237.

5'-O-(4,4'-Dimethoxytrityl)-2'-O,4'-propylene-5-methyluridine (11). Receipt 11 was carried out according to the method described in Example 8 with the formation of 11 as a colorless amorphous solid (quantitative yield).

Range13C-NMR (125 MHz, CDCl3): 11,83, 25,03, 31,46, 55,26, 66,10, 68,18, 70,64, 80,89, 86,94, 89,38, 91,37, 110,14, 113,36, 123,81, 127,18, 128,09, 128,13, 130,10, 135,17. 135,19, 135,37, 136,13, 144,22, 149,99, 158,74, 163,80.

Range1H-NMR (400 MHz, CDCl3): to 1.32 (3H, s). to 1.67 (1H, m) of 1.76 (1H, m) and 1.83 (2H, m), 2,73 (1H, broad), 3,37 (2H, s), 3,79 (6N, C) 4,06 (1H, m), 4,20 (1H, m), 4,30 (1H, q, j 6.6 Hz), 4.72 in (1H, q, j 6.6 Hz), of 5.99 (1H, s), at 6.84 (4H, m), and 7.3 to 7.7 (N, m), 8,44 (1H, s), 8,61 (1H, m).

IR(KBR)max: 3403, 2951, 1688, 1509, 1252 cm-1.

FAB-MS (mNBA): 601 [M+H]>/p>51-O-(4,4'-Dimethoxytrityl)-2'-O,4'-propylene-5-methyluridine-3'-O(2-cyanoethyl-N,N-aminobutiramida)phosphoritic (12). The connection 12 was carried out according to the method described in example 9, to obtain the 12 in the form of a colorless amorphous solid (quantitative yield).

Range1H-NMR (400 MHz, CDCl3): of 1.20 (3H, s) to 1.22 (3H, s) of 1.29 (3H, s) is 1.31 (3H, s) of 1.78 (3H, s), 1,5-2,4 (6N, m), 2,6-2,8 (2H, m), 3,6-3,8 (4H, m), 3,80 (6N, (C), a 4.03 (1H, m), 4,13 (1H, m), 4,36 (1 H, d, 6.6 Hz), to 4.98 (1H, d, 6.6 Hz), 6,04 (1H, s), 6.8 or 6.9 (4H, m), 7,3-7,5 (N), TO $ 7.91 (1H, d, 1.5 Hz), of 8.25 (1H, broad).

FAB-MS (mNBA):801[M+H]+.

(C) Determining the ability of oligonucleotides to form a duplex with a complementary chain.

TPL(melting temperature) was measured by the method of Comparative example 1, presented in the description. From the data presented in Table 5 it follows that the values of TPL, duplexes of the oligonucleotide With respect to the oligonucleotide (complementary RNA) or oligonucleotide F (complementary DNA) (-0,5 or -2,0With per modification) have almost the same high values as duplex nucleotide D (natural DNA).

(d) Determination of resistance to the action of nucleases

USTOIChIVOSTI of example 2, listed in the description. The oligonucleotides in the amount of 26 μg/ml was added in a solution containing 50 mm Tris-HCl (pH 8.0) and 10 mm MgCL2, after which the solution was added 7,14 µg/ml snake venom phosphodiesterase. Resistance to nucleases was determined as a percentage of the amount of remaining oligonucleotides and their original number. The results obtained are presented in Table 6.

Discussion

As follows from the data presented in Table 5, the ability of the oligonucleotide With the present invention to the formation of a duplex with a complementary chain was so high that the ability of natural DNA. As can be seen from Table 6, in contrast to the oligonucleotide a, which is not detected after 10 minutes of incubation, the solution is still present 94% of the oligonucleotide Century As you can see, oligonucleotide In the present invention has a significantly higher resistance to nucleases than the known oligonucleotide A. Extremely high resistance to nucleases compounds of the present invention is not obvious to a person skilled in this field.

Industrial applicability

Oligonucleotide analogues and nucleos the ski drugs possessing good stability, as detection probes specific genes as primers for the amplification or as intermediate compounds for their preparation.

Claims

1. Nucleoside analog of formula (I):

in which R1represents a hydrogen atom or a group protecting the hydroxyl;

R2represents a hydrogen atom, a group protecting the hydroxyl group of phosphoric acid, a protected group, phosphoric acid or a group of the formula P(R3R4in which R3and R4are the same or different and represent a hydroxyl group, a protected hydroxyl group, alkoxygroup containing 1 to 4 carbon atoms, allylthiourea containing 1 to 4 carbon atoms, cyanoacetylurea containing 1 to 5 carbon atoms, amino group, substituted alkyl group containing 1 to 4 carbon atoms;

But alkylenes group containing 1 to 4 carbon atoms; a represents a substituted purine-9-ilen group or substituted 2-oxo-pyrimidine-1-ilen group containing at least one Deputy, selected from hydroxyl group, carbon; or its salt.

2. Nucleoside analogue p. 1 or its salt, in which R1represents a hydrogen atom, acetyl group, benzoyloxy group, benzyl group, p-methoxybenzyloxy group, dimethoxytrityl group, monomethoxypolyethylene group or tert-butyldiphenylsilyl group.

3. Nucleoside analogue according to any one of p. 1 or 2, or its salt, in which R2represents a hydrogen atom, acetyl group, benzoyloxy group, benzyl group, parameterising group, tert-butyldiphenylsilyl group, R(OS2H4SP)(N(CH3)2), -P(och3)(N(CH3)2), fastonline group, or a group of 2-chlorpheniramine acid or group, or group 4-chlorpheniramine acid.

4. Nucleoside analogue according to any one of paragraphs.1-3 or its salt, in which a represents a methylene group.

5. Nucleoside analogue according to any one of paragraphs.1-4 or its salt, which is a 6-aminopurin-9-yl-protected amino group, 2-amino-6-hydroxypurine-9-yl, 2-amino-6-hydroxypurine-9-yl-protected amino group, 2-amino-6-hydroxypurine-9-yl-protected amino group and a hydroxyl group, 2-oxo-4-amino-pyrimidine-1-yl, 2-oxo-4-amino-pyrimidine-1-yl to secure midin-1-yl or 4-amino-5-methyl-2-oxo-pyrimidine-1-yl-protected amino group.

6. Nucleoside analogue according to any one of paragraphs.1 - 5 or its salt, which is a 6-benzylaminopurine-9-ilen, adreniline, 2-isobutylamino-6-hydroxypurine-9-ilen, guanidino, 2-oxo-4-benzoylpiperidine-1-ilen, casinillo, 2-oxo-5-methyl-4-benzoylpiperidine-1-ilen, 5-methylcytosine, brazilrio or ciminillo group.

7. Nucleoside analog or its salt selected from the group of:

2'-O,4'-C-teleguinin,

2'-O,4'-C-utilizationin,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-6-N-benzyladenine,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-2-N-isobutylamine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamine

2'-O,4'-C-ethylene-2-N-isobutylamine,

2'-O,4'-C-ethylene-6-N-benzyladenine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-6-N-benzyladenine-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-2-N-isobutylamino-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide,

2'-O,4'-C-ethyleneimine,

2'-O,4'-C-ethylene-5-methyluridine,

2'-O,4'-C-atransitive,

2'-O,4'-C-ethylene-5-methylcytidine,

3',5'-di-O-benzyl-2'-O,4'-C-ethyleneimine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethyleneimine,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-5-mutilationcelecrity-2'-O,4'-C-ethylene-4-N-benzoylation,

3',5'-di-O-benzyl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine,

2'-O,4'-C-ethylene-4-N-benzoylation,

2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylenedi-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-5-methyluridine-3'-O-(2-cyanate-N,N-aminobutiramida)phosphoamide,

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoylation-3'-O-(2-Tianeti-N,N-aminobutiramida)phosphoamide and

5'-O-dimethoxytrityl-2'-O,4'-C-ethylene-4-N-benzoyl-5-methylcytidine-3'-O-(2-Tianeti N,N-aminobutiramida)phosphoamide.

8. Oligonucleotide analogue comprising one or more structures of the formula (2)

in which a represents alkylenes group having 1 to 4 carbon atoms;

Represents a substituted purine-9-ilen group or substituted 2-oxo-pyrimidine-1-ilen group containing at least one Deputy, selected from hydroxyl groups, protected hydroxyl groups, amino groups, protected amino groups, alkyl groups containing 1 to 4 carbon atoms; or its pharmacologically acceptable salt.

9. Oligonucleotide analogue under item 8, or its pharmacologically acceptable salt, koteski acceptable salt, which is a 6-aminopurin-9-yl, 6-aminopurin-9-yl-protected amino group, 2-amino-6-hydroxypurine-9-yl, 2-amino-6-hydroxypurine-9-yl-protected amino group, 2-amino-6-hydroxypurine-9-yl-protected amino group and a hydroxyl group, 2-oxo-4-amino-pyrimidine-1-yl, 2-oxo-4-amino-pyrimidine-1-yl-protected amino group, 2-oxo-4-hydroxy-pyrimidine-1-yl, 2-oxo-4-hydroxy-5-methyl-pyrimidine-1-yl 4-amino-5-methyl-2-oxo-pyrimidine-1-yl or 4-amino-5-methyl-2-oxo-pyrimidine-1-yl-protected amino group.

11. Oligonucleotide analogue under item 8 or 9, or its pharmacologically acceptable salt, which is a 6-benzylaminopurine-9-yl, adenine, 2-isobutylamino-6-hydroxypurine-9-yl,guanine, 2-oxo-4-benzoylpiperidine-1-yl, cytosine, 2-oxo-5-methyl-4-benzoylpiperidine-1-yl, 5-methylcytosine, urally or teminal.

12. Oligonucleotide analogue according to any one of paragraphs.8-11 or its pharmacologically acceptable salt as a medicinal product that has antisensitivity or antigenic activity.

 

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