RU2559873C2 - Mimetics of poly (adp-ribose) and process for producing thereof - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to PAR mimetics and method of obtaining thereof for the synthesis of novel medical drugs of formula

where Y - nucleotide residue, amine derivative of an aliphatic compound, fluorescence dye; Z - nucleoside residue, (k+1)·m = 1-200; X is O or S; R₁ and R₂ are residues of a disacharide nucleoside or a residue of formulas

wherein n = 2-6; 2-6 or 1-4 respectively, N = nucleoside residue, or -((CH₂)_nO)_m-(P = X(OH))O-N-, wherein n = 2-6, m = 1-6, R₃ is a branching agent of formula

wherein N′- disacharide nucleoside residue, n - a number up to 100, or residues of formulas

n = 2-6; or

n = 2-6; or

n = 1-4,

wherein B = adenine-9-yl, uracyl-1-yl, cytosine-1-yl or guanine-9-yl.

EFFECT: novel PAR mimetics, maintaining all key functional groups and spaces between them, required for the reaction with proteins, wherein the said mimetics are modified for the purpose of synthesis and stability.

2 cl, 21 ex, 2 tbl, 10 dwg

Description

The invention relates to molecular biology, medical chemistry, biochemistry and biotechnology, specifically to means for regulating the metabolism of poly (ADP-ribose) (PAR), which are polymer derivatives of disaccharide nucleosides and their analogues, including their linear and branched copolymers with nucleosides and their derivatives.

A disaccharide nucleoside is a nucleoside with an additional carbohydrate residue attached. An additional residue is connected to the pentafuranose cycle of the nucleoside by an O-glycosidic bond. The disaccharide nucleoside analogs of the present invention are nucleosides modified at the 2 ′, 3 ′ or 5 ′ position with O-hydroxyalkyl, O-hydroxyalkylmethyl, N-hydroxyalkylcarboxamide, O-hydroxyalkyl (thio) phosphoryl or O-polyalkylene glycol (thio) phosphoryl fragments.

Poly-ADP-ribosylation is a post-translational modification of proteins in eukaryotic cells. The process is catalyzed by poly (ADP-ribose) polymerases (PARP). These enzymes convert nicotinamide adenine dinucleotide (NAD ⁺ ) to poly (ADP-ribose) (PAR) with the release of nicotinamide.

The reviews (D.D ′ Amours, et al. Biochem J. 342, 249-268 1999; P.O. Hassa, et al., Microbiol. Mol. Biol. Rev. 70, 789-829, 2006) examined the diverse biological role of this polymer. To date, the reaction mechanism of poly (ADP-ribosylation) has not been studied in detail. The active center of the enzyme is quite extended, it binds the NAD + substrate and the “primer” poly (ADP-ribose) attached to the protein. The key stage is the formation of an O-glycosidic bond, with the formation of nicotinamide. Multipoint fixation of NAD + on nicotinamide, pyrophosphate, and adenosine residues orientes the 2′-O-hydroxyl group to attach to the C′-1 carbon atom to release nicotinamide (Fig. 1).

PAR is known to be involved in the modulation of chromatin structure, replication, transcription, DNA repair and cell differentiation. A detailed study of the intracellular functions of poly (ADP-ribose) continues to this day. Polymer molecules, both isolated from natural sources and synthesized in vitro using poly (ADP-ribose) polymerase, can contain more than 200-300 monomeric units. Linear sections with a length of 20-50 units alternate with branched fragments, which, apparently, stabilize the conformation of poly (ADP-ribose). The structure of poly (ADP-ribose) was determined by NMR spectroscopy and enzymatic hydrolysis with snake venom phosphodiesterase and alkaline phosphatase, which led to diphosphate α-D-ribofuranosyl- (1 ″ → 2 ′) adenosine and the corresponding disaccharide nucleoside α- RibAdo, the structure of which was determined by NMR spectroscopy (AM Ferro, NJOppenheimer Proc. Natl. Acad. Sci. USA. 75, 809-813, 1978). The structure of the trisaccharide nucleoside located at the branching point is also confirmed by physicochemical methods (T. Sugimura, M. Miwa Mol. Cell. Biochem. 138, 5-12, 1994).

The degradation of poly (ADP-ribose) involves ADP-ribosylprotein lyase and poly (ADP-ribose) -glycohydrolase (SAR). ADP-ribosylprotein lyase is responsible for hydrolysis of the bond between the protein and the nearest residue of ADP-ribose. PARG, with exo- and endoglycosidase activity, cleaves the bonds between adenosine and the ribofuranose residue to form ADP ribose (D'Amours D., et al Biochem J. 342, 249-268, 1999), therefore, the protein is covalently bound to the cell STEAM and free polymer. Recently, it was shown that free PAR, entering the cytoplasm from the nucleus, is toxic and can cause cell death (Y. Wang, V.L. Dawson, T.M. Dawson. Exp Neurol. 218, 193-202, 2009).

Poly (ADP-ribosylation) proteination is catalyzed by several polymerases, which are constantly and in significant quantities in the cell. All of them use NAD ⁺ as a substrate and transfer ADP-ribose to glutamate and lysine residues in proteins. PARP-2, PARP-3, as well as tankyrase 1 and 2, etc., were found in eukaryotic cells (R. Krishnakumar and WL Kraus. Molecular Cell 39, 6-24, 2010). Of the enzymes that carry out poly (ADP-ribosylation), PARP-1 is the main one, and it is he who is responsible for the synthesis of 90% poly (ADP-ribose) in the cell. This enzyme specifically recognizes DNA breaks, activates and synthesizes PAR, covalently attached to certain proteins, the enzyme itself being the main acceptor. Poly (ADP-ribosylation) proteination is considered as the main mechanism providing the formation of an intracellular signal about DNA damage and modulation of protein functions in response to genotoxic effects. Excessive activation of PARP during massive DNA breaks, greatly reducing the content of intracellular NAD +, leads to suppression of glycolysis and mitochondrial respiration and causes cell death according to the variant of necrosis. (M.V. Sukhanov, O.I. Lavrik, S.N. Khodyreva, Mol. Biol. 38, 834-847, 2004).

To date, no chemical methods for the synthesis of poly (ADP-ribose) have been developed (E.V. Efimtseva, I.V. Kulikova, S.N. Mikhailov, Molecular Biology, 43, 327-338, 2009) and only recently synthesized its structural element 2′-O-α-D-ribofuranosyladenosine (SN Mikhailov et al Tetrahedron, 64, 2871-2876, 2008). Based on disaccharide nucleosides, new effective regulators of biosynthesis of PAR can be created.

There are several ways to regulate the metabolism of poly (ADP-ribose) associated with suppression (inhibition of PARP, activation of PARG) or, conversely, with the activation of its biosynthesis (activation of PARP, inhibition of PARG). In recent years, there has been an active search for poly (ADP-ribose) polymerase (PARP) inhibitors, which are considered promising target enzymes for creating drugs in the treatment of stroke, ischemia, diabetes, arthritis, colitis and other inflammatory diseases. PARP inhibitors can also be used as potential anticancer and antiviral agents (P. Jagtap, C. Szabo, Nature Rev. Drug Discovery, 4, 421-440, 2005). The main attention of researchers is focused on nicotinamide analogues. It was shown that 3-aminobenzamide inhibits PARP at concentrations of 10 ^-6 M. Further studies have led to the discovery of potent heterocyclic inhibitors with nanomolar inhibition constants. A number of promising compounds are undergoing clinical trials: in combination with alkylating agents in antitumor therapy, as well as for the treatment of heart attack (DV Ferraris. J. Med. Chem. 53, 4561-4584, 2010). The use of alkylating agents in chemotherapy of cancer leads to DNA damage in both tumor and normal cells, and the DNA repair system is activated. The use of PARP inhibitors in complex chemotherapy will reduce the concentration of alkylating agents and, therefore, reduce the overall toxicity of the treatment. A search is also being made for poly (ADP-ribose) -glycohydrolase (PARH) inhibitors, an enzyme responsible for the hydrolysis of PAR in the cell and determining its level (DW Koh, et al. J. Med. Chem. 46, 4322-4332, 2003).

Currently, seven heterocyclic PARP inhibitors, which are analogues of nicotinamide, are undergoing clinical trials (D.V. Ferraris. J. Med. Chem. 53, 4561-4584, 2010). The disadvantages of existing PARP-1 inhibitors include, first of all, the poor solubility of heterocyclic compounds in water, as well as their short lifetime.

In the series of nucleosides, PARP inhibitors are 2′-deoxynucleosides (AD Pivazyan et al., Biochem. Pharm., 44, 947-953, 1992; Preiss et al., FEBS Lett., 19, 244-246, 1971) and disaccharide nucleosides (S.Η. Khodyreva, O. I. Lavrik, A. L. Zakharenko, S. N. Mikhailov, I. V. Kulikova, E. V. Efimtseva. Means for inhibiting the enzyme poly (ADP-ribose) polymerase 1 person RF Patent 2411948). Natural 2′-deoxynucleosides are structural elements of nucleic acids and participate in numerous processes in the cell, which excludes the specificity of their effect on a particular enzyme. Disaccharide nucleosides are more potent PARP-1 inhibitors with IC ₅₀ 0.20 - 0.05 mM, however, it is obvious that this level of inhibition is not sufficient to create drugs based on them.

The second way to regulate PAR metabolism is to activate its biosynthesis. In the cell nucleus, the PARP enzyme is activated by binding to damaged DNA. The binding of poly (ADP-ribose) to the PARP self-modification domain (AMD-PARP) provides regulation of the catalytic activity of the enzyme (R. Krishnakumar et al. Molecular Cell., 39, 8-24, 2010). Single-stranded oligodeoxynucleotides have diverse biological activity unrelated to the formation of complementary complexes with nucleic acids (M. Markosian and R.Μ. Hyde, Antiviral Chem. Chemotherapy 16, 91-102, 1995). Single-stranded guanosine-rich oligodeoxynucleotides cause cell death by caspase activation mechanism (US Patent Application 2009/0312399), have pronounced anti-HIV activity (US Patent 6323185, US Patent 6355785). Oligonucleotides containing thiophosphate groups prevent the penetration of viruses through the cell membrane (D.I. Bernstein et al, Antimicrob.Agents Chemotherapy, 52, 2727-2733, 2008; T. Matsumara et al, Gastroenterology, 137, 673-681, 2009). Polymers are described in which 1,2-dioxipropyl derivatives of nucleic bases are bound by phosphodiester bonds (US Patent 6448373). However, in these compounds the formal distances between the monomer residues are substantially less than in PAR.

The objective of the invention is to obtain mimetics of PAR having the following distinctive properties:

1. In modified PAR, all the main functional groups and the distances between them are retained, which is necessary to ensure potential interactions with proteins; According to the calculated data, poly (ADP-ribose) and its mimetics have significant conformational mobility and the formal distances between C1'-C1 'in two adjacent nucleoside residues are 12-14 Å.

2. It is promising to obtain PAH mimetics in which the pyrophosphate group is replaced by a phosphate residue. In this case, you can use the modified technology of automatic oligonucleotide synthesis using phosphoramidite derivatives. This technology allows the inclusion of commercially available phosphoramidite derivatives of nucleosides, as well as residues of ethylene glycol, propane and butanediols, hydrophobic and fluorescent groups to increase stability and efficient delivery of PAR analogues to the cell. In this case, one or more modified residues can be located in a given position of the OH chain.

3. The protective groups used in the synthesis of disaccharide nucleosides and their derivatives should be combined with the groups used for automatic synthesis, which will significantly reduce the time to obtain PAR mimetics; selective blocking of five hydroxyl groups (two primary and three secondary groups) in 2′-O-α-D-ribofuranosyladenosine and its analogs is an important step in the preparation of phosphoramidite derivatives.

4. To increase the stability of PAR mimetics to PARH, it is proposed to change the configuration of the O-glycosidic bond in the residues of 2′-O-α-D-ribofuranosyladenosine or use derivatives thereof, such as 2′-O-pentafuranosyladenosine and 2′-O-hydroxyalkoxymethyl nucleosides. It is also possible to obtain various copolymers based on these compounds.

PAR mimetics, binding to PARP and activating it, can replace PAR in various cellular processes (modulation of chromatin structure and repair of damaged DNA, transcription, cell differentiation, etc.) and are of significant interest as prototypes of new drugs for molecular medicine, as well as new objects in research in molecular biology, medical chemistry and biochemistry. It is important to note that to date, structural PAR mimetics are not described in the literature and are not used in industry.

The task of obtaining mimetics PAR with the specified characteristics is solved by the fact that as the claimed compounds are proposed linear and (or) branched polymers of the general formula

where ribo- and 2′-deoxyribonucleosides attached to the polymer by a phosphodiester bond (Υ: deoxyadenosin-5′-yl, deoxyguanosin-5′-yl, deoxycytidin-5′-yl, deoxythymidine- can be used as terminal fragments Υ, Ζ 5′-yl; Ζ: adenosin-3′-yl, guanosin-3′-yl, cytidin-3′-yl, uridin-3′-yl), an amino derivative of a branched aliphatic compound (C7-6-amino (2- hydroxymethyl) -hexan-1-yl), triphenylmethane derivatives (Tr - triphenylmethyl, MMTr - 4-methoxytriphenylmethyl, DMTr - 4,4′-dimethoxytriphenylmethyl) fluorescent dyes triphenylmeth ananova series (fluorescein, eosin, rhodamine) associated with the mimetic ester bond, or an amide bond with a C7 linker attached to the mimetic. The total length of the main chain of the polymer (k + 1) * m can vary from 1 to 100, and the phosphodiester groups (X = O) can be combined or completely replaced with thiophosphoryl (X = S). As monomer units R ₁ and R ₂ , disaccharide nucleosides 2′-O-β-D-ribofuranosyladenosine or 2′-O-β-D-ribofuranosylguanosine or 2′-O-β-D-ribofuranosylcytidine or 2′- can be used O-β-D-ribofuranosyluridine or 2′-O-α-D-arabinofuranosyladenosine or 2′-O-α-D-arabinofuranosylguanosine or 2′-O-α-D-arabinofuranosylcytidine or 2′-O-α-D- arabinofuranosyluridine or 2′-O-α-L-arabinofuranosyladenosine or 2′-O-α-L-arabinofuranosylguanosine or 2′-O-α-L-arabinofuranosylcytidine or 2′-O-α-L-arabinofuranosyluridine or 2′-O -α-D-ribofuranosyladenosine or 2′- O-α-D-ribofuranosylguanosine or 2′-O-α-D-ribofuranosylcytidine or 2′-O-α-D-ribofuranosyluridine or 2′-O-β-D-glucopyranosyladenosine or 2′-O-β-D- glucopyranosylguanosine or 2′-O-β-D-glucopyranosylcytidine or 2′-O-β-D-glucopyranosyluridine, interconnected by 5′-5 ″ phosphodiester bonds, or their analogs of the general formula:

where B = Ade (adenine-9-yl); B = Ura (uracil-1-yl); B = Cyt (cytosin-1-yl); B = Gua (guanine-9-yl).

Also, fragments of the general formula - ((CH ₂ ) _n O) _m - (P = X (OH)) ON-, where n = 1-6, m = 1-6, can be used as monomer units R ₁ and R ₂ , and N = the remainder of the nucleoside, or the General formula

где N = остаток нуклеозида.

where N = nucleoside residue.

R ₃ splitters _{of the} general formula can be introduced into the main chain of polymers - mimetics PAR

где N′ остаток дисахаридного нуклеозида - 2′-O-β-D-рибофуранозиладенозина или 2′-O-β-D-рибофуранозилгуанозина или 2′-O-β-D-рибофуранозилцитидина или 2′-O-β-D-рибофуранозилуридина, соединенного с основной цепью миметика фосфодиэфирными связями по 3' и 5′ положениям аденозинового фрагмента, и с боковой цепью по 5′′-гидроксильной группе β-D-рибофуранозного остатка; или

where N ′ is the residue of a disaccharide nucleoside - 2′-O-β-D-ribofuranosyladenosine or 2′-O-β-D-ribofuranosylguanosine or 2′-O-β-D-ribofuranosylcytidine or 2′-O-β-D-ribofuranosyluridine connected to the main chain of the mimetic by phosphodiester bonds at the 3 ′ and 5 ′ positions of the adenosine moiety, and to the side chain at the 5 ′ ′ hydroxyl group of the β-D-ribofuranose residue; or

(nucleoside units are interconnected by a 3′-5′-phosphodiester bond, the side chain of the mimetic is attached to the main chain through an acyclic moiety - (CH ₂ ) _n ) The monomer units shown are introduced into the polymer as phosphoramidite derivatives.

To obtain polymers, phosphoramidite synthons with monomethoxytrityl (MMTr) or dimethoxytrityl (DMTr) protective groups are used. Compounds 1-6, where B = Ade ^Bz (H ⁶ -benzoyladenin-9-yl); B = Ura (uracil-1-yl); B = Cyt ^Bz (N ⁴ -benzoylcytosin-1-yl); B = Gua ^iBu (N ² -isobutyroylguanin-9-yl). R = MMTr or DMTr were obtained from disaccharide nucleosides (Fig. 2).

In the case of compound 1, the monomethoxytrityl group blocks the 5 ′ ′ - hydroxyl group of the additional 2′-O-β-D-ribofuranose residue. Also, isomeric phosphoramidite derivatives 2 were obtained in which the di (mono) methoxytrityl group blocks the 5′-OH residue of adenosine. In this case, glycosylation was carried out by 1,2,3-tri-O-benzoyl-5-O-levulininyl-O-ribofuranose (SN Mikhailov et al. Chemistry & Biodiversity, 2, 1153-1163, 2005), which significantly reduced the number stages and increase the total yield of products 2. For the synthesis of polymers, phosphoramidite derivatives with monomethoxytrityl and dimethoxytrityl groups were used, acyl groups: benzoyl (ΒZ) and isobutyroyl (iBu) groups were used to protect exocyclic groups of heterocyclic bases.

Phosphoramidite derivatives based on 2′-O-hydroxyalkoxynucleosides 7–9 were synthesized, where B = Ade ^Bz , Ura, Cyt ^Bz , Gua ^iBu , R = MMTr or DMTr, n = 2-4 (Fig. 3).

Along with disaccharide nucleosides and their derivatives, a number of commercially available phosphoramidite synthones 10-14 were used to obtain PAR mimetics, where B = Ade ^Bz , Ura, Cyt ^Bz , Gua ^iBu (Figure 4).

To obtain branched polymers, synthons 15 and 16 were used, where B = Ade ^Bz , Ura, Cyt ^Bz , Gua ^iBu , n = 2-4 (Fig. 5).

Phosphoramidite synthons were included in PAR mimetics using automated oligonucleotide synthesis technology (Fig. 6, 11)

After the synthesis of the polymer on a polymer carrier, the levulinyl group (Lev = C (O) CH ₂ CH ₂ C (O) CH ₃ ) hydrazine hydrate was selectively removed. Subsequent chain extension and removal of the cyanethyl and acyl protecting groups led to branched PAH mimetics (Figure 6).

The synthesis of PAR mimetics is carried out using the above-described phosphoramidite blocks 1-16 in automatic mode on an insoluble silicate, polystyrene or polyacrylate carrier by oligonucleotide chemistry at a temperature of 20 ° C. In the course of automatic synthesis, monomer units are sequentially attached to the 5′- or 5 ′ ′ end of the synthesized oligomer until a polymer of the required length is obtained. Each attachment cycle consists of 4 stages:

1. Release (removal of the triphenylmethyl (MMTr or DMTr) protective group from the 5 ′ hydroxyl group of the oligonucleotide chain with trichloroacetic acid).

2. Condensation (addition of an amidophosphite block in the presence of a catalyst of -1H-tetrazole or 2-ethylthiotetrazole).

3. Capping (acetylation of unreacted hydroxyl groups with a mixture of propionic anhydride and pyridine in a ratio of 1: 1 to 1: 3 in tetrahydrofuran).

4. Oxidation (conversion of internucleotide phosphite to phosphotriether or thiophosphodiester).

At the final stage of the synthesis of polymer mimetics, PAR removes the acyl protecting groups under standard conditions (conc. Aqueous ammonia, 6 hours, 55 ° C).

The previously considered modified phosphoramidites differ from each other both in the reactivity of the amidophosphite group and in the type of trityl protection on the hydroxyl group.

The most reactive are synthons with amidophosphite on the primary hydroxyl group.

The standard protecting group at the 5 ′ position of amidophosphite blocks is 4,4′-dimethoxytrityl. In addition, a monomethoxytrityl protecting group may be used. Its removal rate with a standard deblocking solution (3% trichloroacetic acid in methylene chloride) is lower compared to the dimethoxytrityl group. Its use is advisable in the synthesis of a set of oligomers that differ among themselves by one monomer unit: Ν, Ν-1, Ν-2, etc. Moreover, in the final set, each oligomer will contain a monomethoxytrityl protective group.

Standard approaches in oligonucleotide chemistry allow the introduction of both phosphodiester and thiophosphoryl internucleotide bonds. Thus, thiophosphoryl derivatives of oligomeric PAR mimetics can be obtained.

Automatic synthesis of oligomeric Mimetic PARs allows you to get not only linear but also branched oligomers. This is achieved through the use of appropriate commercially available amidophosphite blocks, for example dendrimer phosphoramidites.

The functionality of the considered PAR mimetics can be significantly expanded by introducing reactive groups, fluorescence labels, fluorescence quenchers, fragments that ensure the transport of oligomers to the target. Amino groups, aldehyde, thiol, phosphate groups, biotin, and an EDTA fragment can act as reactive groups. Fluorescence reporter fragments based on fluorescein derivatives, cyanine dyes, acridine, phenanthrene, pyrene, as well as fluorescence quenchers, for example, based on azobenzene derivatives, can also be introduced into oligomers.

The presence of a hydrophobic dimethoxytrityl group at the 5 ′ end of the synthesized oligomer significantly facilitates the chromatographic isolation of the desired full-sized product. Due to this, PAR oligomeric mimetics can be obtained with a high degree of purification using reverse-phase HPLC.

The developed methodology allows one to obtain regular and irregular structures with a degree of polymerization of up to 100 monomer units, which is comparable to natural PAIRs. The number (k + 1) · m is determined by the technical capabilities of the OH synthesizer. The developed technology allows synthesizing polymers based on diesters of phosphoric acid with a repeating monomer unit, polymers with a regular structure with repeating several units, as well as irregular polymers with several monomer units. A distinctive feature of this technology is the production of polymers with a given molecular weight, and therefore with certain properties.

The present invention is illustrated by examples of the synthesis of phosphoramidite blocks and PAH mimetics with different functional and reporter groups at the ends of the polymer chain and in the middle of the chain.

The structure of the claimed compounds was confirmed by UV, NMR and mass spectrometry. NMR spectra were recorded on a Bruker AMX400 spectrometer (Germany). Chemical shifts (δ) are measured relative to the internal standard tetramethylsilane (δ 0 ppm) and are given in parts per million. The values of the spin – spin interaction constants (J) are measured in hertz. When describing ¹ H-NMR spectra, the following abbreviations were adopted: c — singlet, ush. s - broadened singlet, d - doublet, t - triplet, m - multiplet. Modified oligonucleotides were obtained on an automated synthesizer Applied Biosystems (USA). Thin layer chromatography was performed on Kieselgel 260 F plates (Merck), spots were shown in ultraviolet light (λ = 254 nm). Column chromatography was performed on Kieselgel 60 silica gel (0.063-0.200 mm, Merck). Purification of solvents was carried out according to standard methods.

Example 1. Synthesis of N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D) -ribofuranosyl) -3-O-acetyl-5-O - (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] adenine (1, B = Ade ^Bz , R = MMTr. Fig. 7).

a) N ⁶ -benzoyl-9- [2-O- (β-D) -ribofuranozil) -3,5-O (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D- ribofuranosyl] adenine.

A solution of 1.66 g (1.56 mmol) of N ^6- benzoyl-9- [2-O- (2,3,5-tri-O-benzoyl-β-D-ribofuranosyl) -3,5-O- (1,1, 3,3-tetraisopropyl disiloxane-1,3-diyl) -β-D) ribofuranosyl] adenine (SN Mikhailov et al J. Carbohydr. Chem. 16, 75-92, 1997) in a 0.15 Μ solution of sodium methylate in dry methanol ( 42 ml) was held for 40 minutes. at 20 ° C, 10% acetic acid in methanol was added to pH 7.0, evaporated to dryness in vacuo. The residue was dissolved in 100 ml of dichloromethane and washed successively with 30 ml of a saturated solution of sodium bicarbonate and water (2 × 30 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was chromatographed on a column containing 35 g of silica gel. The material was eluted with 1.5% ethanol / dichloromethane (500 ml). The fractions containing the product were combined and evaporated to dryness in vacuo. Yield 680 mg (58% foam). R _ƒ 0.28 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.28 br. s (1H, NH), 8.76 s (1H, H-8), 8.43 s (1H, H-2), 8.03 d (2H, Bz), 7.60 t (1H, Bz), 7.52 t (2H, Βz) , 6.11 s (1H, H-1 ′ Ado), 5.33 s (1H, H-1 ′ Rib), 4.76 dd (1H, J _{3 ′} , _{2 ′} = 4.6, J _{3 ′, 4} = 7.6, Η-3 ′ Rib), 4.50 dd (1H, J _{3 ′, 2 ′} 4.2, J _{3 ′, 4} = 9.2, H-3 ′ Ado), 4.47 d (1H, H-2 ′ Ado), 4.29 d (1H, J _5′a5′b = _- 13.2, Η-5′a Ado), 4.17 m (2H, Η-2 ′ Rib, Η-4 ′ Ado), 4.03 m (3H, H-4 ′, 5′a Rib, Η-5′b Ado), 3.76 d (1H, J _{5′b, 5′a} = -10.7, Η-5′b Rib), 1.14-0.95 m (28H, i-Pr). ¹³ C-NMR (CDCl ₃ ): 164.58 (C = O), 152.17 (C-2), 149.89 (C-6), 149.66 (C-4), 140.55 (C-8), 132.73, 128.72, and 127.85 (Bz), 123.65 (C-5), 106.11 (C-1 ′, Rib), 89.17 (C-1 ′, Ado), 83.79, 81.67 (C-4 ′), 76.62, 75.49 (C-2 ′) , 68.98, 68.91 (C-3 ′), 60.21 (C-5 ′, Rib), 59.42 (C-5 ′, Ado), 17.32, 17.25, 17.15, 17.07, 16.89, 13.27, 12.78, 12.78 and 12.57 (i -Pr).

b) N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3,5-O- (1,1,3 , 3-tetraisopropyl disiloxane-1,3-diyl) -β-D-ribofuranosyl] adenine.

Prepared from 518 mg (0.7 mmol) of N ^6- Benzoyl-9- [2-O- (β-D-ribofuranosyl) -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl ) -β-D-ribofuranosyl] adenine according to the standard procedure for the administration of a trityl protective group followed by acylation. The product was chromatographed on a column containing 30 g of silica gel. The substance was eluted with a 1.5% ethanol / dichloromethane system. The fractions containing the product were combined and evaporated to dryness in vacuo. Yield 640 mg (83% foam). R _ƒ 0.41 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 8.98 br. s (1H, NH), 8.59 s (1H, H-8), 8.02-7.15 m (18H, H-2, Bz, Ph), 6.74 d (2H, Ph), 5.76 s (1H, H-1 ′ Ado), 5.73 dd (1H, J _{3 ′, 2 ′} 3.3, J _{3 ′, 4 ′} 6.9, Η-3 ′ Rib), 5.54 s (1H, H-1 ′ Rib), 5.53 d (1H, Η- 2 ′ Rib), 5.11 dd (1H, J _{3 ′, 2 ′} 4.8, J _{3 ′, 4 ′} 8.8, H-3 ′ Ado), 4.87 d (1H, H-2 ′ Ado), 4.27 ddd (1H, J _{4 ′, 5′a} 4.8, J _{4 ′, 5′b} 3.7, H-4 ′ Rib), 4.05 m (3H, H-4 ′, 5′a, 5′b Ado), 3.73 s (3H, OMe), 3.36 dd (1H, _{J5′a, 5′b} - 10.3, Η-5′a Rib), 3.23 dd (1H, Η-5′b Rib), 2.09 s (3H, Ac), 2.03 s ( 3H, Ac), 1.11-1.04 m (28H, i-Pr). ¹³ C-NMR (CDCl ₃ ): 169.75, 169.42, 164.56 (C = O), 158.70 (Ph), 152.63 (C-2), 150.60 (C-6), 149.42 (C-4), 142.44 (C- 8), 135.06, 132.75, 130.45, 128.90, 128.47, 128.38, 127.81 and 127.02 (Bz, Ph), 123.36 (C-5), 113.03 (Ph), 105.45 (C-1 ′, Rib), 88.84 (C- 1 ′, Ado), 86.17 (Ph ₃ C), 81.22, 79.82 (C-4 ′), 78.64, 74.97 (C-2 ′), 71.36, 69.96 (C-3 ′), 62.95 (C-5 ′, Rib), 59.85 (C-5 ′, Ado), 55.15 (OMe), 20.46 and 20.37 (Ac), 17.33, 17.28, 17.18, 17.00, 16.82, 13.23, 12.95, 12.84 and 12.60 (i-Pr).

c) N ⁶ -Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -β-D-ribofuranosyl] adenine.

To a solution of 324 mg (0.29 mmol) N ^6- benzoyl-9- [2-O (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3,5-O- ( 1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D-ribofuranosyl] adenine in tetrahydrofuran (4 ml) was added 280 mg (0.88 mmol) of tetrabutylammonium fluoride trihydrate in tetrahydrofuran (3 ml). This reaction takes place for 15 minutes at 20 ° C. The reaction mixture was evaporated in vacuo and then reevaporated with chloroform (2 × 10 ml). The residue was chromatographed on a column containing 30 g of silica gel, eluting with 1.5% ethanol / dichloromethane. The fractions containing the product were combined and evaporated to dryness in vacuo. Yield 230 mg (92% as a foam), R _ƒ 0.30 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.11 g (1H, NH), 8.77 s (1H, H-8), 8.01-7.28 m (18H, H-2, Bz, Ph), 6.88 d (2H, PhOMe) , 5.90 d (1H, J _{1 ′, 2 ′} = 7.0, H-1 ′ Ado), 5.42 dd (1H, J _{3 ′, 2 ′} = 5.0, J _{3 ′, 4 ′} = 5.2, Η-3 ′ Rib ), 5.35 dd (1H, J _{2 ′, 1 ′} = 2.2, Η-2 ′ Rib), 5.01 dd (1H, J _{2 ′, 3 ′} = 4.8, H-2 ′ Ado), 4.81 d (1H, H -1 ′ Rib), 4.64 dd (1H, J _{3 ′, 4 ′} = 1.1, H-3 ′ Ado), 4.29 dd (1H, H-4 ′ Ado), 4.04 ddd (1H, J _{4 ′, 5 ′ a} = 3.6, J _{4 ′, 5′b} = 4.2, H-4 ′ Rib), 3.96 d (1H, J _{5′a, 5′b} - = 13.2, H-5′a Ado), 3.73 s (3H , OMe), 3.70 dd (1H, Η-5′b Ado), 3.32 dd (1H, J _5′a5′b = -10.6, H-5′a Rib), 3.23 dd (1H, Η-5′b Rib), 2.05 s (3H, Ac), 2.02 s (3H, Ac). ¹³ C-NMR (CDCl ₃ ): 169.90, 164.53 (C = O), 158.92 (Ph), 152.12 (C-2), 150.36 (C-6), 149.78 (C-4), 141.42 (C-8) , 134.79, 132.94, 130.48, 128.96, 128.47, 128.11, 127.87 and 127.32 (Bz, Ph), 124.16 (C-5), 113.39 (Ph), 106.62 (C-1 ′, Rib), 89.72 (C-1 ′ , Ado), 86.93 (Ph ₃ C), 86.83 (C-4 ′), 81.04, 76.36 (C-2 ′), 75.00, 72.02 (C-3 ′), 71.14 (C-5 ′, Rib), 63.01 (C-5 ′, Ado), 55.21 (OMe), 20.43, 19.87 (Ac).

d) N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -5-O-tert-butyldiphenylsilyl-β-D- ribofuranosyl] adenine.

To a solution of 370 mg (0.43 mmol) N ⁶ -benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -β-D-ribofuranosyl] adenine and imidazole 35 mg (0.51 mmol) in dry pyridine (4 ml) was added 0.19 ml (0.75 mmol) of tert-butyldiphenylsilyl chloride, kept for 48 h at 20 ° C. The mixture was evaporated to dryness, the residue was dissolved in chloroform (20 ml) and washed with water (10 ml), saturated sodium bicarbonate solution (10 ml) and then with water (10 ml). The organic layer was dried with anhydrous sodium sulfate, filtered, evaporated to dryness, re-evaporated with toluene (2 × 10 ml). The residue was chromatographed on a column containing 20 g of silica gel. The substance was eluted with 1% ethanol / dichloromethane (400 ml). The fractions containing the product were combined and evaporated to dryness in vacuo. Yield 420 mg (88%, foam). R _ƒ 0.37 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.21 us (1H, NH), 8.60 s (1H, H-8), 8.04-7.18 m (28H, H-2, Bz, Ph), 6.83 d (2H, PhOMe) , 6.13 d (1H, J _{1 ′, 2 ′} = 2.8, H-1 ′ Ado), 5.49 dd (1H, J _{3 ′, 2 ′} 5.0, J _{3 ′, 4 ′} = 5.2, Η-3 ′ Rib) , 5.46 dd (1H, J _{2 ′, 1 ′} = 2.4, H-2 ′ Rib), 5.20 d (1H, H-1 ′ Rib), 4.95 dd (1H, J _{2 ′, 3 ′} = 5.6, H- 2 ′ Ado), 4.76 dd (1H, J _{3 ′, 4 ′} = 5.9, H-3 ′ Ado), 4.22 d (1H, H-4 ′ Rib), 4.05 ddd (1H, J _{4 ′, 5′a} = 3.6, J _{4 ′, 5′b} = 3.4, Η-4 ′ Ado), 4.03 d (1H, J _{5′a, 5′b} - = 11-2, Η-5′a Ado), 3.86 d ( 1H, Η-5′b Ado), 3.74 s (3H, OMe), 3.36 dd (1H, J _{5′a, 5′b} = -9.9, Η-5′a Rib), 3.30 dd (1H, Η- 5′b Rib), 2.09 s (3H, Ac), 2.06 s (3H, Ac), 1.04 s (9H, t-Bu). ¹³ C-NMR (CDCl ₃ ): 165.94, 162.53 (C = O), 157.92 (Ph), 151.12 (C-2), 151.26 (C-6), 148.76 (C-4), 143.41 (C-8) , 134.79, 132.94, 130.48, 128.96, 128.47, 128.11, 127.87 and 127.32 (Bz, Ph), 124.16 (C-5), 113.39 (Ph), 107.42 (C-1 ′, Rib), 88.41 (C-1 ′ , Ado), 87.05 (Ph ₃ C), 84.42, 81.52 (C-4 ′), 81.29, 76.36 (C-2 ′), 75.00, 71.80 (C-3 ′), 71.14 (C-5 ′, Rib) 63.08 (C-5 ′, Ado), 55.34 (OMe), 26.97 (t-Bu), 20.43, 19.43 (Ac).

d) N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3-O-acetyl-5-O-tert- butyl diphenylsilyl-β-D-ribofuranosyl] adenine.

To a solution of 350 mg (0.32 mmol) of N ^6- benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -5-O-tert- butyldiphenylsilyl-β-D-ribofuranosyl] adenine in dry pyridine (20 ml) was added 50 μl (0.5 mmol) of acetic anhydride and kept for 2 h at 20 ° C, after which 0.3 ml of methanol was added and kept for 30 min. The reaction mixture was evaporated in vacuo. The residue was dissolved in chloroform (30 ml), washed with 10% aqueous sodium bicarbonate solution (15 ml), water (2 × 10 ml). The organic layer was dried over anhydrous sodium sulfate, filtered, evaporated, and then evaporated with toluene (2 × 10 ml). The residue was chromatographed on a column containing 20 g of silica gel. The substance was eluted with a 1.5% ethanol / dichloromethane system. The fractions containing the product were combined and evaporated to dryness in vacuo. Yield 277 mg (76% foam). R _ƒ 0.40 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.06 g (1H, NH), 8.54 s (1H, H-8), 8.04-7.18 m (28H, H-2, Bz, Ph), 6.76 d (2H, PhOMe) , 6.10 d (1H, J _{1 ′, 2 ′} = 3.8, H-1 ′ Ado), 5.43 dd (1H, J _{3 ′, 2 ′} = 4.9, J _{3 ′, 4 ′} = 5.3, H-3 ′ Ado ), 5.34 dd (1H, J _{2 ′ , 4 ′} = 5.6, H 3 ′ Rib), 5.32 dd (1H, J _{2 ′, 1 ′} = 1.4, H-2 ′ Rib ), 5.09 d (1H, J- _{1 ′, 2 ′} = 3.8, H-1 ′ Ado), 4.99 d (1H, H-1 ′ Rib), 4.28 d (1H, H-4 ′ Ado), 4.10 ddd ( 1H, J _{4 ′, 5′a} = 3.6, J _{4 ′, 5′b} = 3.4, H-4 ′ Rib), 3.96 d (1H, J _{5′a, 5′b} - = 11.2, Η-5 ′ a Ado), 3.86 d (1H, Η-5′b Ado), 3.73 s (3H, OMe), 3.36 m (2H, Η-5′a, Η-5′b Rib), 2.11 s (3H, Ac ), 2.05 s (3H, Ac), 2.02 s (3H, Ac), 1.05 s (9H, t-Bu). ¹³ C-NMR (CDCl ₃ ): 165.94, 162.53 (C = O), 157.92 (Ph), 151.12 (C-2), 151.26 (C-6), 148.76 (C-4), 143.41 (C-8) , 134.79, 132.94, 130.48, 128.96, 128.47, 128.11, 127.87 and 127.32 (Bz, Ph), 124.16 (C-5), 113.39 (Ph), 107.42 (C-1 ′, Rib), 88.41 (C-1 ′ , Ado), 87.05 (Ph ₃ C), 84.42, 81.52 (C-4 ′), 81.29, 76.36 (C-2 ′), 75.00, 71.80 (C-3 ′), 71.14 (C-5 ′, Rib) 63.08 (C-5 ′, Ado), 55.34 (OMe), 26.97 (f-Bu), 20.43, 20.87, 19.43 (Ac).

f) N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3-O-acetyl-β-D-ribofuranosyl] adenine.

To a solution of 250 mg (0.22 mmol) N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3-O-acetyl- 5-O-tert-butyldiphenylsilyl-β-D-ribofuranosyl] adenine in tetrahydrofuran (5 ml) was added 105 mg (0.33 mmol) of tetrabutylammonium fluoride trihydrate in tetrahydrofuran (5 ml) and kept for 15 min at 20 ° C. The reaction mixture was evaporated in vacuo, re-evaporated with chloroform (2 × 10 ml). The residue was chromatographed on a column containing 20 g of silica gel. The material was eluted with 5% ethanol / dichloromethane (500 ml). The fractions containing the product were combined and evaporated to dryness in vacuo. Yield 177 mg (89% foam). R _ƒ 0.34 in the 5% ethanol / dichloromethane system. ¹ H NMR (CDCl ₃ ): 9.31 cf (1H, NH), 8.87 s (1H, H-8), 8.01-7.28 m (18H, H-2, Bz, Ph), 6.79 d (2H, PhOMe), 5.83 d (1H, J _{1 ′, 2 ′} = 5.2, H-1 ′ Ado), 5.45 dd (1H, J _{2 ′, 1 ′} = 2.2, Η-2 ′ Rib), 5.32 dd (1H, J _{3 ′ , 2 ′} = 5.0, J _{3 ′, 4 ′} = 4.2, Η-3 ′ Rib), 5.21 dd (1H, H-1 ′ Rib), 5.06 dd (1H, J _{2 ′, 3 ′} = 6.8, H- 2 ′ Ado), 4.75 dd (1H, J _{4 ′, 4 ′} = 3.1, H-3 ′ Ado), 4.29 d (1H, H-4 ′ Rib), 4.04 ddd (1H, J _{4 ′, 5′a} = 2.6, J _{4 ′, 5′b} = 3.2, H-4 ′ Ado), 3.96 d (1H, J _{5′a, 5′b} - = 11.2, Η-5′a Ado), 3.84 d (1H, Η-5′b Ado), 3.73 s (3H, OMe), 3.35 dd (1H, J _{5′a, 5′b} = -10.2, Η-5′a Rib), 3.28 dd (1H, Η-5 ′ b Rib), 2.11 s (3H, Ac), 2.07 s (3H, Ac), 2.01 s (3H, Ac). ¹³ C-NMR (CDCl ₃ ): 169.90, 167.57, 164.53 (C = O), 158.92 (Ph), 153.12 (C-2), 151.36 (C-6), 148.78 (C-4), 141.35 (C- 8), 134.79, 132.94, 130.48, 128.96, 128.47, 128.11, 127.87 and 127.32 (Bz, Ph), 123.15 (C-5), 113.43 (Ph), 106.72 (C-1 ′, Rib), 88.76 (C- 1 ′, Ado), 87.95 (Ph ₃ C), 81.88 (C-4 ′), 81.04, 76.36 (C-2 ′), 75.00, 72.02 (C-3 ′), 71.14 (C-5 ′, Rib) 63.01 (C-5 ′, Ado), 55.76 (OMe), 21.15, 20.46, 19.97 (Ac).

g) N ^6- Benzoyl-9- [2-O (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3-O-acetyl-5-O- (2- cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] adenine (1, B = Ade ^Bz , R = MMTr).

To a solution of 150 mg (0.17 mmol) N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl) -3-O-acetyl- β-D-ribofuranosyl] adenine in 10 ml of dichloroethane, diisopropylethylamine (0.08 ml, 0.5 mmol) and 2-cyanoethyl-Ν, Ν-diisopropylchlorophosphoramidite (0.07 ml, 0.35 mmol) were added. After 2 hours, a 10% aqueous sodium hydrogen carbonate solution (3 ml) was added, and stirred for 10 minutes. A solution of sodium bicarbonate (20 ml) and dichloromethane (20 ml) were added. The organic layer was separated, washed with water (2 × 20 ml), dried with anhydrous sodium sulfate, filtered, evaporated to dryness. The residue was chromatographed on a column containing 15 g of silica gel. The material was eluted with ethyl acetate / hexane / triethylamine (70/29/1).

The fractions containing the product were combined, evaporated to dryness in vacuo. Yield 140 mg (74% foam). R _ƒ 0.42 in the 5% ethanol / dichloromethane system. ³¹ P-NMR (CDCl ₃ ): 151.47, 150.12.

Example 2. Synthesis of N ^6- Benzoyl-9- [2-O- (2,3-di-O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl) -3-O-acetyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] adenine (1, B = Ade ^Bz , R = DMTr).

The substance was obtained analogously to example 1 with a total yield of 17%. R _ƒ 0.44 in the 5% ethanol / dichloromethane system. ³¹ P-NMR (CDCl ₃ ): 151.45, 150.11.

Example 3. Synthesis of N ^6- Benzoyl-9- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] - 3-O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl} adenine (2, B = Ade ^Bz , R = DMTr. Fig. 8).

a) N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-β-D-ribofuranosyl) -3-O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl] adenine.

From 595 mg (0.73 mmol) N ⁶ -Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -β-D-ribofuranosyl] adenine (SN Mikhailov et al Chemistry & Biodiversity, 2, 1153-1163, 2005) obtained N ⁶ -Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-β-D ribofuranosyl) -3-O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl] adenine according to the standard tritylation-acylation method. N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -3-O-acetyl-5-O-dimethoxytrityl-β- Without preliminary chromatographic purification, D-ribofuranosyl] adenine was dissolved in 10 ml of pyridine, cooled to 0 ° C and 1.89 ml (33 mmol, 45 equivalents with respect to the starting N ⁶ -Benzoyl-9- [2- (9- (2, 3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -β-D-ribofuranosyl] adenine) acetic acid Next, 0.214 ml (4.41 mmol, 6 equivalents to the starting material) were added dropwise at 0 ° C. N ⁶ -benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulinyl-β-D-ribofuranosyl) -β-D-ribofuranosyl] adenine) g Drazine monohydrate and kept for 40 min at 0 ° C. The reaction mixture was diluted with ethyl acetate (30 ml) and washed successively with saturated aqueous sodium bicarbonate (20 ml) and water (4 × 20 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated in vacuo. The residue was chromatographed on a column containing 15 g of silica gel. The material was eluted with ethyl acetate / hexane / triethylamine (66/33/1 by volume). The product yield in 3 stages 578 mg (75%) as a white foam. R _ƒ 0.26 in the system ethyl acetate / hexane / triethylamine (49/49/2 by volume). ¹ H-NMR (DMSO-D ₆ ): 11.29 ss (1H, ΝΗ), 8.68 s (1Η, H-8), 8.65 s (1H, H-2), 8.10-7.20 m (24H, Bz, Ph) , 6.88 d (2H, J = 3.5, mPhOMe), 6.86 d (2H, J = 3.3, mPhOMe), 6.40 d (1H, J _{1 ′, 2 ′} = 5.4, H-1 ′ Ado), 5.59 dd (1H , J _{2 ′, 3 ′} = 6.0, J _{3 ′, 4 ′} = 4.6, H-3 ′ Ado), 5.56-5.54 m (2H, Η-2 ′, H-3 ′ Rib), 5.46 dd (1H, J _{1 ′, 2 ′} = 5.4, J _{2 ′, 3 ′} = 6.0, H-2 ′ Ado), 5.31 d (1H, J _{4 ′, 3′a} = 1.2, Н-1 ′ Rib), 5.12 t ( 1H, J _{5 ′, OH} = 5.2, 5′-OH), 4.37 ddd (1H, J _{4 ′, 5′a} = 4.3, J _{5′a, 5′b} = -9.0, Η-5′a Ado) , 4.35-4.30 m (1H, Η-5′b Ado), 3.79 s (6H, OMe), 3.42-3.38 m (2H, Η-4 ′ Ado, Η-4 ′ Rib), 3.36-3.32 m (2H , Η-5′a, Η-5′b Rib), 2.07 s (3H, Ac). ¹³ C-NMR (DMSO-D ₆ ): 170.15 (C = O-Ac), 166.19, 165.28, 164.81 (C = O-Bz), 158.44 (Ph), 152.25 (C-2), 152.01 (C-6 ), 150.73 (С-4), 144.94 (С-8), 144.23, 135.69, 135.62, 134.28, 134.14, 133.56, 132.95, 130.02, 129.50, 129.09, 128.91, 128.77, 128.16, 127.93, 127.14 (Bz, DMTr) 126.04 (C-5), 113.48 (m-MeOPh), 105.60 (C-1 ′, Rib), 87.52 (Ph ₃ C), 86.10 (C-1 ′, Ado), 82.59, 81.19 (C-41) , 76.89, 75.14 (С-2 ′), 72.40, 71.15 (С-3 ′), 63.09, 61.24 (С-5 ′, Ado), 55.34 (OMe), 20.58 (Ac).

b) N ^6- Benzoyl-9- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] -3- O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl} adenine (2, B = Ade ^Bz , R = DMTr).

To a solution of 500 mg (0.47 mmol) of N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-β-D-ribofuranosyl) -3-O-acetyl-5-O-dimethoxytrityl- β-D-ribofuranosyl] adenine and 36 mg (0.52 mmol) of 1H-tetrazole in 6 ml of acetonitrile / pyridine (5/1 by volume) were added dropwise 0.254 ml (0.80 mmol) of 2-cyanoethyl-N, N, N ′, N '-Tetraisopropylphosphoramidite. The reaction was carried out in an inert atmosphere of nitrogen. After 1 h, a 10% aqueous sodium hydrogen carbonate solution (3 ml) was added, and stirred for 10 minutes. A solution of sodium bicarbonate (20 ml) and ethyl acetate (20 ml) were added. The organic layer was separated, washed with water (2 × 20 ml), dried with anhydrous sodium sulfate, filtered, evaporated to dryness. The residue was chromatographed on a column containing 15 g of silica gel. The material was eluted with ethyl acetate / hexane / triethylamine (69/29/2). The fractions containing the product were combined, evaporated to dryness in vacuo. Yield 502 mg (85% foam). R _ƒ 0.66 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.92, 150.69.

Example 4. Synthesis of N ^6- Benzoyl-9- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl) -N, N-diisopropylamidophosphinyl-β-D-ribofuranosyl] - 3-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl} adenine (2, B = Ade ^Bz , R = MMTr).

The substance was obtained according to example 3, starting from N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -β-D-ribofuranosyl ] adenine. The total yield of the product in four stages is 67%. R _ƒ 0.65 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.91, 150.67.

Example 5. Synthesis of 1- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] -3-O-acetyl -5-O-dimethoxytrityl-β-D-ribofuranosyl} uracil (2, B = Ura, R = DMTr).

The substance was obtained according to example 3, starting from 1- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -β-D-ribofuranosyl] uracil. The total yield of the product in four stages is 71%. R _ƒ 0.75 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.90, 150.65.

Example 6. Synthesis of 1- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -β-D-ribofuranosyl] -3-O-acetyl -5-O-monomethoxytrityl-β-D-ribofuranosyl} uracil (2, B = Ura, R = MMTr).

The substance was obtained according to example 3, starting from 1- [2-O (2,3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -β-D-ribofuranosyl] uracil. The total yield of the product in four stages is 77%. R _ƒ 0.75 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.87, 150.61.

Example 7. Synthesis of N ^6- Benzoyl-9- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -α-D-arabinofuranosyl] - 3-O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl} adenine (3, B = Ade ^Bz , R = DMTr).

The substance was obtained according to example 3, starting from N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-α-D-arabinofuranosyl) -β-D-ribofuranosyl ] adenine. The total yield of the product in four stages is 63%. R _ƒ 0.67 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.90, 150.70.

Example 8. Synthesis of N ^6- Benzoyl-9- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -α-D-arabinofuranosyl] - 3-O-acetyl-5-O-monomethoxytrityl-β-D-ribofuranosyl} adenine (3, Β = Αde ^Βz , R = MMTr).

The substance was obtained according to example 3, starting from N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-α-D-arabinofuranosyl) -β-D-ribofuranosyl ] adenine. The total yield of the product in four stages is 63%. R _ƒ 0.67 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.90, 150.71.

Example 9. Synthesis of N ^6- Benzoyl-9- {2-O- [2,3-di-O-benzoyl-5-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -α-D-ribofuranosyl] - 3-O-acetyl-5-O-dimethoxytrityl-β-D-ribofuranosyl} adenine (5, B = Ade ^Bz , R = DMTr).

The substance was obtained from 2′-O-α-D-ribofuranosyladenosine (SN Mikhailov et al Tetrahedron, 64, 2871-2876, 2008). R _ƒ 0.70 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.87, 150.68.

Example 10. Synthesis of N ^6- Benzoyl-9- {2-O - [(3- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxypropoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β- D-ribofuranosyl} adenine (7, B = Ade ^Bz , R = MMTr, n = 3. Fig. 9).

a) 9- {2-O - [(3-Acetoxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) -β-D-ribofuranosyl} adenine.

, Н-4′), 4.01 д (1Н, Н-5′b), 3.83-3.78 м (1Н, $$OC{H}_{2}C{H}_{2}C\underset{\_}{H}HOAc$$

), 3.70-3.65 м (1H, $$OC{H}_{2}C{H}_{2}CH\underset{\_}{H}OAc$$

), 2.01 с (3Н, ОАс), 1.93-1.89 м (2Н, ОСН₂СН₂СН₂ОАс), 1.10-0.88 м (28Н, i-Pr). ¹³С-ЯМР (CDCl₃): 171.12 (С=O), 155.59 (С-6), 153.20 (С-8), 149.28 (С-4), 138.76 (С-2), 120.41 (С-5), 95.15 (OCH₂O), 89.96 (С-1′), 81.67 (С-4′), 78.45 (С-2′), 69.29 (С-3′), 65.13 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OAc$$

), 61.78 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OAc$$

), 60.10 (С-5′), 29.00 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 21.02 (ОАс), 17.59, 17.49, 17.44, 17.29, 17.16, 17.04, 13.54, 13.10, 13.03, 12.83 (i-Pr).To a solution containing 1.02 g (2 mmol) of 9- [3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) -β-D-ribofuranosyl] adenine and 0.76 g (4 mmol) ) 3- (hydroxymethoxy) propan-1-ol diacetate in 10 ml of 1,2-dichloroethane under nitrogen atmosphere with stirring and cooling to -10 ° C, 0.749 ml (6.4 mmol) of tin tetrachloride was added. The solution was held for 40 minutes at -10 ° C. To the solution was added 20 ml of a saturated solution of sodium bicarbonate and 30 ml of dichloromethane, stirred for 20 minutes and filtered through Hyflo Super-cel (celite). The organic layer was separated, washed with water (2 × 50 ml), dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo. The residue was chromatographed on a column containing 35 g of silica gel. The substance was eluted with a 1.5% ethanol / dichloromethane system. Yield 0.73 g (56%) as a white foam. R _ƒ 0.23 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 8.30 s (1H, H-8), 8.12 s (1H, H-2), 6.05 s (1H, H-1 ′), 5.66 s (2H, NH ₂ ), 4.98 s (2H, OCH ₂ O), 4.73 dd (1H, J _{3 ′, 2 ′} = 4.4, J _{3 ′, 4 ′} = 9.3, Η-3 ′), 4.50 d (1H, H-2 ′), 4.23 d (1H, J _{5′a, 5′b} = -13.2, Η-5′a), 4.12-4.04 m (3H, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OAc$$

, Н-4 ′), 4.01 d (1Н, Н-5′b), 3.83-3.78 m (1Н, $$OC{H}_{2}C{H}_{2}C\underset{\_}{H}HOAc$$

), 3.70-3.65 m (1H, $$OC{H}_{2}C{H}_{2}CH\underset{\_}{H}OAc$$

), 2.01 s (3H, OAc), 1.93-1.89 m (2H, OCH ₂ CH ₂ CH ₂ OAc), 1.10-0.88 m (28H, i-Pr). ¹³ C-NMR (CDCl ₃ ): 171.12 (C = O), 155.59 (C-6), 153.20 (C-8), 149.28 (C-4), 138.76 (C-2), 120.41 (C-5) , 95.15 (OCH ₂ O), 89.96 (С-1 ′), 81.67 (С-4 ′), 78.45 (С-2 ′), 69.29 (С-3 ′), 65.13 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OAc$$

), 61.78 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OAc$$

), 60.10 (С-5 ′), 29.00 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 21.02 (OAc), 17.59, 17.49, 17.44, 17.29, 17.16, 17.04, 13.54, 13.10, 13.03, 12.83 (i-Pr).

b) 9- {2-O - [(3-Hydroxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) -β-D-ribofuranosyl} adenine.

), 4.92 d (1Н, $$OCH\underset{\_}{H}O$$

), 4.65 дд (1H, J_3′,2′=4.4, J_3′,4′=9.3, Η-3′), 4.40 д (1Н, Η-2′), 4.25 д (1Η, J_5′a5′b=-13.4, Η-5′а), 4.02 дд (1H, J_4′,5′b=2.5, H-4′), 3.67 дд (1H, Η-5′b), 3.77-3.62 м (4H, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 1.84-1.82 м (2H, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OH$$

), 1.24-0.97 м (28Н, i-Pr). ¹³С-ЯМР (CDCl₃): 155.63 (С-6), 152.84 (С-8), 148.72 (С-4), 138.74 (С-2), 120.15 (С-5), 95.24 (OCH₂O), 88.73 (С-1′), 81.57 (С-4′), 78.71 (С-2′), 69.11 (С-3′), 65.57 ( $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 59.90 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OH$$

), 59.30 (С-5′), 32.18 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 17.58, 17.42, 17.27, 17.12,17.00, 13.50, 13.01, 12.74 (i-Pr).9- {2-O - [(3-Acetoxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D-ribofuranosyl} adenine (1.38 g , 2.16 mmol) was treated with 15 ml of a 4M solution of methylamine in ethanol (MeNH ₂ / EtOH) and left for a day at room temperature. After 24 hours, the contents of the flask were evaporated in vacuo, re-evaporated with dichloromethane (2 × 15 ml) and applied to a column containing 30 g of silica gel. The substance was eluted with 5% ethanol / dichloromethane. Yield 1.10 g (85% foam). R _ƒ 0.19 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 8.27 s (1H, H-2), 8.22 s (1H, H-2), 6.33 s (2H, NH ₂ ), 6.08 s (1H, H-1 ′), 5.00 d (1H, J = -7.0, $$OC\underset{\_}{H}HO$$

), 4.92 d (1H, $$OCH\underset{\_}{H}O$$

), 4.65 dd (1H, J _{3 ′, 2 ′} = 4.4, J _{3 ′, 4 ′} = 9.3, Η-3 ′), 4.40 d (1H, Η-2 ′), 4.25 d (1Η, J _{5 ′ a5′b} = -13.4, Η-5′a), 4.02 dd (1H, J _{4 ′, 5′b} = 2.5, H-4 ′), 3.67 dd (1H, Η-5′b), 3.77-3.62 m (4H, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 1.84-1.82 m (2H, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OH$$

), 1.24-0.97 m (28Н, i-Pr). ¹³ C-NMR (CDCl ₃ ): 155.63 (C-6), 152.84 (C-8), 148.72 (C-4), 138.74 (C-2), 120.15 (C-5), 95.24 (OCH ₂ O) , 88.73 (С-1 ′), 81.57 (С-4 ′), 78.71 (С-2 ′), 69.11 (С-3 ′), 65.57 ( $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 59.90 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OH$$

), 59.30 (С-5 ′), 32.18 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 17.58, 17.42, 17.27, 17.12.17.00, 13.50, 13.01, 12.74 (i-Pr).

c) N ^6- Benzoyl-9- {2-O - [(3-hydroxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D ribofuranosyl} adenine.

), 4.96 д (1Н, $$OCH\underset{\_}{H}O$$

), 4.70 дд (1Н, J_3′,2′-=4.1, J_3′,4′=9.3, Η-3′), 4.49 д (1H, Η-2′), 4.25 д (1Η, J_5′а,5′b=-13.5, Η-5′а), 4.17 д (1Н, Η-4′), 4.04-3.99 м (2Η, $$OC\underset{\_}{H}HC{H}_{2}C{H}_{2}OH$$

, Н-5′b), 3.73-3.66 м (3Н, $$OCH\underset{\_}{H}C{H}_{2}C{\underset{\_}{H}}_{2}OH$$

), 1.83-1.81 м (2Н, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OH$$

), 1.01-0.99 м (28Н, i-Pr). ¹³С-ЯМР (CDCl₃): 164.97 (С=O), 152.74 (С-6), 150.83 (С-8), 149.82 (С-4), 141.31 (С-2), 133.90, 132.82, 128.91, 127.48 (Ph), 123.68 (С-5), 95.30 (OCH₂O), 89.03 (С-1′), 81.79 (С-4′), 78.54 (С-2′), 69.31 (С-3′), 66.08 ( $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 60.02 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OH$$

), 59.97 (С-5′), 32.18 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 17.57, 17.41, 17.48, 17.28, 17.13, 17.00, 13.52, 13.03, 12.80 (i-Pr).To a suspension of 9- {2-O - [(3-Hydroxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) -β-D-ribofuranosyl} adenine ( 0.75 g, 1.26 mmol) in dry pyridine (30 ml) was added trimethylchlorosilane (0.8 ml, 6.33 mmol). The suspension was stirred at room temperature for 40 minutes. Benzoyl chloride (0.8 ml, 6.89 mmol) was added, left at 20 ° C and stirring for 2 hours. The reaction mixture was cooled to 0 ° C, water (10 ml) was added, and after 5 min 25% aqueous ammonia solution (10 ml) was added. The mixture was kept for 30 min at 20 ° C and evaporated in vacuo. Evaporated with toluene (2 × 15 ml). The residue was dissolved in ethyl acetate (50 ml), the resulting solution was washed successively with saturated sodium bicarbonate solution (20 ml), water (2 × 20 ml), dried over anhydrous sodium sulfate, filtered, evaporated to dryness in vacuo. The residue was chromatographed on a column containing 30 g of silica gel. The substance was eluted with a 2% ethanol / dichloromethane system. Yield 0.64 g (72%) as a white foam. R _ƒ 0.30 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.32 s (1H, NHBz), 8.77 s (1H, H-8), 8.38 s (1H, H-2), 8.03-7.42 m (5H, Bz), 6.14 s ( 1H, H-1 ′), 5.02 d (1H, J = -6.9, $$OC\underset{\_}{H}HO$$

), 4.96 d (1H, $$OCH\underset{\_}{H}O$$

), 4.70 dd (1H, J _{3 ′, 2 ′} - = 4.1, J _{3 ′, 4 ′} = 9.3, Η-3 ′), 4.49 d (1H, Η-2 ′), 4.25 d (1Η, J _{5 ′ A, 5′b} = -13.5, Η-5′a), 4.17 d (1H, Η-4 ′), 4.04-3.99 m (2Η, $$OC\underset{\_}{H}HC{H}_{2}C{H}_{2}OH$$

, H-5′b), 3.73-3.66 m (3H, $$OCH\underset{\_}{H}C{H}_{2}C{\underset{\_}{H}}_{2}OH$$

), 1.83-1.81 m (2H, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OH$$

), 1.01-0.99 m (28H, i-Pr). ¹³ C-NMR (CDCl ₃ ): 164.97 (C = O), 152.74 (C-6), 150.83 (C-8), 149.82 (C-4), 141.31 (C-2), 133.90, 132.82, 128.91, 127.48 (Ph), 123.68 (С-5), 95.30 (OCH ₂ O), 89.03 (С-1 ′), 81.79 (С-4 ′), 78.54 (С-2 ′), 69.31 (С-3 ′) 66.08 ( $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 60.02 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OH$$

), 59.97 (С-5 ′), 32.18 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 17.57, 17.41, 17.48, 17.28, 17.13, 17.00, 13.52, 13.03, 12.80 (i-Pr).

g) N ⁶ -Benzoyl-9- {2-O - [(3-levulinyloxypropoxy) methyl] -3,5- (9- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β- D-ribofuranosyl} adenine.

), 4.93 д (1Н, $$OCH\underset{\_}{H}O$$

), 4.70 дд (1H, J_3′,2′=4.5, J_3′,4′=8.9, Η-3′), 4.48 д (1Н, Η-2′), 4.17 д (1Η, J_5′a,5′b=-13.1, Η-5′а), 4.00 м (3Н, Η-4′, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 3.99 д (1H, Η-5′b), 3.78-3.66 м (2Н, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.69 т (2Н, J=5.6, CH₂-Lev), 2.51 т (2H, CH₂-Lev), 2.12 с (3Н, СН₃-Lev), 1.90-1.86 м (2Н, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

), 1.07-0.99 м (28Н, i-Pr). ¹³С-ЯМР (CDCl₃): 206,53 172.66, 164.97 (С=O), 152.53 (С-6), 150.91 (С-8), 149.62 (С-4), 141.27 (С-2), 133.77, 132.70, 128.77, 127.46 (Ph), 123.58 (С-5), 95.12 (OCH₂O), 89.04 (С-1′), 81.69 (С-4′), 78.21 (С-2′), 69.22 (С-3′), 65.01 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 61.77 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 59.89 (С-5′), 37.03( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 29.80 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 28.88 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 27,92 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

), 17.47, 17.40, 17.31, 17.18, 17.04, 16.92, 13.52, 13.41, 12.96 (i-Pr).To a solution containing 0.64 g (0.92 mmol) of N ^6- Benzoyl-9- {2-O - [(3-hydroxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1, 3-diyl) -β-D-ribofuranosyl} adenine, 0.27 g (2.3 mmol) of levulinic acid in 15 ml of 1,2-dichloroethane at 20 ° C was added 0.76 g (3.6 mmol) of N, N′-dicyclohexylcarbodiimide. The solution was kept for 1.5 hours at 20 ° C. The precipitate was filtered off. The organic layer was separated, washed with water (2 × 50 ml), dried over anhydrous sodium sulfate, filtered and evaporated in vacuo. The residue was chromatographed on a column containing 35 g of silica gel. The substance was eluted with a 1.5% ethanol / dichloromethane system. Yield 0.57 g (78%) as a white foam. R _ƒ 0.34 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.41 s (1H, NHBz), 8.72 s (1H, H-8), 8.31 s (1H, H-2), 8.01-7.44 m (5H, Bz), 6.10 s ( 1H, H-1 ′), 4.96 d (1H, J = -6.7, $$OC\underset{\_}{H}HO$$

), 4.93 d (1H, $$OCH\underset{\_}{H}O$$

), 4.70 dd (1H, J _{3 ′, 2 ′} = 4.5, J _{3 ′, 4 ′} = 8.9, Η-3 ′), 4.48 d (1H, Η-2 ′), 4.17 d (1Η, J _{5 ′ a, 5′b} = -13.1, Η-5′a), 4.00 m (3H, Η-4 ′, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 3.99 d (1H, Η-5′b), 3.78-3.66 m (2Н, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.69 t (2H, J = 5.6, CH ₂ -Lev), 2.51 t (2H, CH ₂ -Lev), 2.12 s (3H, CH ₃ -Lev), 1.90-1.86 m (2H, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

), 1.07-0.99 m (28H, i-Pr). ¹³ C-NMR (CDCl ₃ ): 206.53 172.66, 164.97 (C = O), 152.53 (C-6), 150.91 (C-8), 149.62 (C-4), 141.27 (C-2), 133.77 , 132.70, 128.77, 127.46 (Ph), 123.58 (С-5), 95.12 (OCH ₂ O), 89.04 (С-1 ′), 81.69 (С-4 ′), 78.21 (С-2 ′), 69.22 ( S-3 ′), 65.01 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 61.77 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 59.89 (С-5 ′), 37.03 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 29.80 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 28.88 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 27.92 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

), 17.47, 17.40, 17.31, 17.18, 17.04, 16.92, 13.52, 13.41, 12.96 (i-Pr).

d) N ⁶ -Benzoyl-9- {2-O - [(3-levulinyloxypropoxy) methyl] -β-D-ribofuranosyl} adenine.

), 4.58 д (1H, $$OCH\underset{\_}{H}O$$

), 4.43 дд (1Н, J_3′,4′=1.3, Η-3′), 4.32 д (1Н, Н-4′), 4.00 м (2Н, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 3.99 д (1H, J_5′a,5′b=-11.7, Η-5′а), 3.94 д (1H, Н-5′b), 3.58-3.37 м (2Н, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.70 т (2Н, J=5.6, CH₂-Lev), 2.51 т (2Η, CH₂-Lev), 2.14 с (3Н, СН₃-Lev), 1.71-1.67 м (2Н, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

). ¹³С ЯМР (CDCl₃): 207.05, 172.82, 164.90 (С=O, 152.33 (С-6), 150.83 (С-8), 150.37 (С-4), 143.32 (С-2), 133.53, 133.04, 128.44, 128.15 (Ph), 124.23 (С-5), 96.16 (OCH₂O, 89.51 (С-1′, 87.83 (С-4′), 80.55 (С-2′), 71.61 (С-3′), 65.46 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 63.03 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 60.32 (С-5′), 38.03 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 29.80 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 28.87 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 27.99 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

).1.42 g (1.78 mmol) N ⁶ -Benzoyl-9- {2-O - [(3-levulinyloxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) β-D-ribofuranosyl} adenine was treated with 2.5 ml of a 0.5 Μ solution of tetrabutylammonium fluoride trihydrate in tetrahydrofuran and kept for 10 min at 20 ° C. The reaction mixture was evaporated in vacuo, re-evaporated with chloroform (2 × 10 ml). The residue was chromatographed on a column containing 30 g of silica gel. The substance was eluted with 4% ethanol / dichloromethane. Yield 803 mg (81%) as a white amorphous R / 0.35 foam in a 10% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.53 ss (1H, NHBz), 8.76 s (1H, H-8), 8.22 s (1H, H-2), 8.03-7.48 m (5H, Bz), 6.03 d ( 1H, J _{1 ′, 2 ′} = 7.2, H-1 ′), 4.94 dd (1H, J _{2 ′, 3 ′} = 4.7, Η-2 ′), 4.67 d (1H, J = -6.5, $$OC\underset{\_}{H}HO$$

), 4.58 d (1H, $$OCH\underset{\_}{H}O$$

), 4.43 dd (1Н, J _{3 ′, 4 ′} = 1.3, Η-3 ′), 4.32 d (1Н, Н-4 ′), 4.00 m (2Н, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 3.99 d (1H, J _{5′a, 5′b} = -11.7, Η-5′a), 3.94 d (1H, H-5′b), 3.58-3.37 m (2Н, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.70 t (2H, J = 5.6, CH ₂ -Lev), 2.51 t (2Η, CH ₂ -Lev), 2.14 s (3H, CH ₃ -Lev), 1.71-1.67 m (2H, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

) ¹³ C NMR (CDCl ₃ ): 207.05, 172.82, 164.90 (C = O, 152.33 (C-6), 150.83 (C-8), 150.37 (C-4), 143.32 (C-2), 133.53, 133.04, 128.44, 128.15 (Ph), 124.23 (C-5), 96.16 (OCH ₂ O, 89.51 (C-1 ′, 87.83 (C-4 ′), 80.55 (C-2 ′), 71.61 (C-3 ′) , 65.46 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 63.03 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 60.32 (S-5 ′), 38.03 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 29.80 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 28.87 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 27.99 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

)

e) N ^6- Benzoyl-9- {2-O - [(3-hydroxypropoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β-D-ribofuranosyl} adenine.

), 6.35 д (1Н, J_1′,2′=6.4, Н-1′), 5.78 дд (1H, J_3′,2′=5.6, J_3′,4′=2.8, Η-3′), 5.33 дд (1H, Η-2′), 4.77 д (1Η, J=-7.2, $$OC\underset{\_}{H}HO$$

), 4.60 д (1H, $$OCH\underset{\_}{H}O$$

), 4.54 ддц (1Н, J_4′,5′a,=3.7, J_4′,5′b=4.1, Η-4′), 3.76 с (3Н, OMe), 3.60 дд (1H, J_5′a,5′b=-10.9, Η-5′а), 3.56 дд (1Н, Н-5′b), 3.45 м (2Н, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 3.30 м (2Н, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OH$$

), 1.57-1.51 м (2Н, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OH$$

). ¹³С-ЯМР (CDCl₃): 165.81, 165.07 (С=O), 158.91 ( $$p\underset{\_}{Ph}OMe$$

), 152.79 (С-6), 152.29 (С-8), 149.76 (С-4), 143.32 (С-2), 141.85, 141.98, 135.82, 133.04, 129.05, 128.57, 128.07, 127.33 (Ph), 123.67 (С-5), 113.45 ( $$m\underset{\_}{Ph}OMe$$

), 95.55 ( $$O\underset{\_}{C}{H}_{2}O$$

), 87.42 ( $$P{h}_3\underset{\_}{C}$$

), 86.65 (С-1′), 82.72 (С-4′), 77.47 (С-2′), 72.39 (С-3′), 66.30 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OH$$

), 63.47 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OH$$

), 59.77 (С-5′), 55.33 (ОМе), 33.91 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

).A solution of 450 mg (0.81 mmol) of N ⁶ -Benzoyl-9- {2-O - [(3-levulinyloxypropoxy) methyl] -β-D-ribofuranosyl} adenine and 426 mg (1.38 mmol) of monomethoxytrityl chloride in dry pyridine (5 ml) kept for 16 hours at 20 ° C. The reaction mixture was cooled to 0 ° C and 0.140 ml (1.21 mmol) of benzoyl chloride was added with stirring. Benzoylation was carried out at 0 ° C, monitoring the course of the reaction by TLC until the complete disappearance of the starting material. Then methanol (2 ml) was added to the reaction mixture and kept for 30 min at room temperature, then it was diluted with dichloromethane (20 ml) and washed successively with saturated aqueous sodium bicarbonate solution (20 ml) and water (2 × 20 ml). The organic layer was dried over anhydrous sodium sulfate, filtered, evaporated. The residue was dissolved in 10 ml of pyridine, cooled to 0 ° C and 2.08 ml (36.4 mmol, 45 equivalents with respect to N ⁶ -Benzoyl-9- {2-O - [(3-levulinyloxypropoxy) methyl] -β-D- ribofuranosyl} adenine) acetic acid. Then 0.236 ml (4.86 mmol, 6 equivalents with respect to the starting nucleoside) of hydrazine monohydrate were added and kept for 1 h at 0 ° C. The reaction mixture was diluted with ethyl acetate (30 ml) and washed successively with saturated aqueous sodium bicarbonate (20 ml), water (4 × 20 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated in vacuo. The residue was chromatographed on a column containing 15 g of silica gel. The substance was eluted with a 2% ethanol / dichloromethane system. The output in 3 stages is 406 mg (60%) as a white foam. R _f 0.35 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 9.31 s (1H, NHBz), 8.73 s (1H, H-8), 8.30 s (1H, H-2), 8.12-7.22 m (22H, Ph), 6.81 d ( 2H, $$m\underset{\_}{Ph}OMe$$

), 6.35 d (1H, J _{1 ′, 2 ′} = 6.4, H-1 ′), 5.78 dd (1H, J _{3 ′, 2 ′} = 5.6, J _{3 ′, 4 ′} = 2.8, Η-3 ′) , 5.33 dd (1Η, Η-2 ′), 4.77 d (1Η, J = -7.2, $$OC\underset{\_}{H}HO$$

), 4.60 d (1H, $$OCH\underset{\_}{H}O$$

), 4.54 dd (1H, J _{4 ′, 5′a} , = 3.7, J _{4 ′, 5′b} = 4.1, Η-4 ′), 3.76 s (3H, OMe), 3.60 dd (1H, J _{5 ′ a, 5′b} = -10.9, Η-5′a), 3.56 dd (1Н, Н-5′b), 3.45 m (2Н, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OH$$

), 3.30 m (2H, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OH$$

), 1.57-1.51 m (2H, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OH$$

) ¹³ C-NMR (CDCl ₃ ): 165.81, 165.07 (C = O), 158.91 ( $$p\underset{\_}{Ph}OMe$$

), 152.79 (С-6), 152.29 (С-8), 149.76 (С-4), 143.32 (С-2), 141.85, 141.98, 135.82, 133.04, 129.05, 128.57, 128.07, 127.33 (Ph), 123.67 (C-5), 113.45 ( $$m\underset{\_}{Ph}OMe$$

), 95.55 ( $$O\underset{\_}{C}{H}_{2}O$$

), 87.42 ( $$P{h}_3\underset{\_}{C}$$

), 86.65 (С-1 ′), 82.72 (С-4 ′), 77.47 (С-2 ′), 72.39 (С-3 ′), 66.30 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OH$$

), 63.47 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OH$$

), 59.77 (С-5 ′), 55.33 (ОМе), 33.91 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

)

g) N ^6- Benzoyl-9- {2-O - [(3- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxypropoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β-D- ribofuranosyl} adenine (7, B = Ade ^Bz , R = MMTr, n = 3).

To a solution of 400 mg (0.48 mmol) N ^6- Benzoyl-9- {2-O - [(3-hydroxypropoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β-D-ribofuranosyl} adenine and 37 mg (0.53 mmol) of 1H-tetrazole in 6 ml of a mixture of acetonitrile / pyridine (5/1 by volume) was added dropwise 0.229 ml (0.72 mmol) of 2-cyanoethyl-N, N, N ′, N′-tetraisopropylphosphoramidite. The reaction was carried out in an inert atmosphere of nitrogen. After 30 minutes, a 10% aqueous solution of sodium hydrogen carbonate (3 ml) was added, and stirred for 10 minutes. A solution of sodium bicarbonate (20 ml) and ethyl acetate (20 ml) were added. The organic layer was separated, washed with water (2 × 20 ml), dried with anhydrous sodium sulfate, filtered, evaporated to dryness. The residue was chromatographed on a column containing 15 g of silica gel. The material was eluted with ethyl acetate / hexane / triethylamine (69/29/2). The fractions containing the product were combined, evaporated to dryness in vacuo. Yield 303 mg (61% foam). R _ƒ 0.35 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (CDCl ₃ ): 150.68, 149.31.

Example 11. Synthesis of N ^6- Benzoyl-9- {2-O - [(2- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxyethoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β- D-ribofuranosyl} adenine (7, B = Ade ^Bz , R = MMTr, n = 2).

The substance was obtained analogously to example 10 with a total yield of 9.8%. R _ƒ 0.34 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (DMSO-D ₆ ): 150.18, 150.14.

Example 12. Synthesis of 1- {2-O - [(2- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxyethoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β-D-ribofuranosyl} uracil (7, B = Ura, R = MMTr, n = 2).

The substance was obtained analogously to example 10 with a total yield of 16%. R _ƒ 0.71 in the ethyl acetate / hexane / triethylamine system (69 / 29.5 / 1.5). ³¹ P-NMR (DMSO-D ₆ ): 150.17, 150.12.

Example 13. Synthesis of 1- {2-O - [(2- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxyethoxy) methyl] -3-O-benzoyl-5-O-dimethoxytrityl-β-D-ribofuranosyl} uracil (7, B = Ura, R = DMTr, n = 2).

The substance was obtained analogously to example 10 with a total yield of 14%. R _ƒ 0.74 in the ethyl acetate / hexane / triethylamine system (69 / 29.5 / 1.5). ³¹ P-NMR (DMSO-D ₆ ): 150.15, 150.09.

Example 14. Synthesis of N ^6- Benzoyl-9- {2-O - [(4- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxybutoxy) methyl] -3-O-benzoyl-5-O-monomethoxytrityl-β- D-ribofuranosyl} adenine (7, B = Ade ^Bz , R = MMTr, n = 4).

The substance was obtained analogously to example 10 with a total yield of 8%. R _ƒ 0.50 in the system ethyl acetate / hexane / triethylamine (69 / 29.5 / 1.5). ³¹ P-NMR (DMSO-D ₆ ): 149.10.

Example 15. Synthesis of N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -3-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -5-O-monomethoxytrityl-β-D -ribofuranosyl} cytosine (15, B = Cyt ^Bz , R = MMTr, n = 3. Fig. 10).

a) N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D ribofuranosyl} cytosine.

), 4.95 д (1H, $$OCH\underset{\_}{H}O$$

), 4.29 д (1Н, J_5′a5′b=-13.7, Η-5′а), 4.25-4.23 м (1Н, Н-4′), 4.22-4.14 м (4Н, Н-2′, H-3′ $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 3.99 д (1Н, Η-5′b), 3.89-3.69 м (2H, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.73 т (2Η, J_CH2-CH2=6.5, CH₂-Lev), 2.56 т (2H, CH₂-Lev), 2.17 с (3H, CH₃-Lev), 1.96-1.88 м (2Н, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

), 1.20-0.85 м (28Н, i-Pr). ¹³С-ЯМР (CDCl₃): 206.99, 177.50, 173.02 (С=O), 162.96 (С-4), 144.81 (С-6), 133.46 (С-2) 133.35, 129.21, 127.20 (Ph), 96.56 (С-5), 94.94 (OCH₂O), 90.64 (С-1′), 82.28 (С-4′), 78.07 (С-2′), 68.12 (С-3′), 65.10 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 62.31 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 59.74 (С-5′), 38.33( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 30.15 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 29.11 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 28.33 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

), 17.79, 17.75, 17.61, 17.38, 17.30, 17.26, 17.16, 13.76, 13.42, 13.21, 12.91 (i-Pr).To a solution containing 0.80 g (1.18 mmol) of N ^4- Benzoyl-1- {2-O - [(3-hydroxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyldisiloxane-1, 3-diyl) -β-D-ribofuranosyl} cytosine (GV Bobkov et al. Tetrahedron, 64, 6238-6251, 2008) and 0.27 g (2.36 mmol) of levulinic acid in 15 ml of dioxane at 20 ° C were added 11 mg (0.091 mmol) Ν, Ν-dimethylaminopyridine (DMAP) and 0.48 g (2.36 mmol) of Ν, Ν-dicyclohexylcarbodiimide. The solution was kept for 1.5 hours at 20 ° C. The precipitated white precipitate of Ν, Ν′-dicyclohexylurea was filtered. The organic layer was separated, washed with water (2 × 50 ml), dried over anhydrous sodium sulfate, filtered and evaporated in vacuo. The residue was chromatographed on a column containing 30 g of silica gel. The substance was eluted with a 1.5% ethanol / dichloromethane system. Yield 0.75 g (82%) as a white foam. R _ƒ 0.43 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 10.20 ss (1H, NHBz), 8.34 d (1H, J _6.5 = 7.5, H-6), 7.97-7.45 m (6H, Bz, H-5), 5.83 s (1H, H-1 ′), 5.07 d (1H, J = -6.5, $$OC\underset{\_}{H}HO$$

), 4.95 d (1H, $$OCH\underset{\_}{H}O$$

), 4.29 d (1Н, J _5′a5′b = -13.7, Η-5′а), 4.25-4.23 m (1Н, Н-4 ′), 4.22-4.14 m (4Н, Н-2 ′, H -3 ′ $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 3.99 d (1H, Η-5′b), 3.89-3.69 m (2H, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.73 t (2Η, J _CH2-CH2 = 6.5, CH ₂ -Lev), 2.56 t (2H, CH ₂ -Lev), 2.17 s (3H, CH ₃ -Lev), 1.96-1.88 m (2Н, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

), 1.20-0.85 m (28Н, i-Pr). ¹³ C-NMR (CDCl ₃ ): 206.99, 177.50, 173.02 (C = O), 162.96 (C-4), 144.81 (C-6), 133.46 (C-2) 133.35, 129.21, 127.20 (Ph), 96.56 (C-5), 94.94 (OCH ₂ O), 90.64 (C-1 ′), 82.28 (C-4 ′), 78.07 (C-2 ′), 68.12 (C-3 ′), 65.10 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 62.31 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 59.74 (С-5 ′), 38.33 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 30.15 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 11/29 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 28.33 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

), 17.79, 17.75, 17.61, 17.38, 17.30, 17.26, 17.16, 13.76, 13.42, 13.21, 12.91 (i-Pr).

b) N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -β-D-ribofuranosyl} cytosine.

), 4.87 д (1Н, $$OCH\underset{\_}{H}O$$

), 4.41 дд (1Н, J_2′3′=5.3, Η-2′), 4.36 дд (1Н, J_3′4′=6.8, Η-3′), 4.16-4.09 м (3Н, H-4′, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 4.06 дд (1H, J_5′a5′b=-12.8, J_5′a4′=2.2, Н-5′а), 3.90 дд (1Н, J_5′a5′b=-12.8, J_5′b4′=1.9, Н-5′b), 3.71-3.62 м (1Н, $$OC{H}_{2}C{H}_{2}C\underset{\_}{H}HOLev$$

), 3.64-3.55 м (1H, $$OC{H}_{2}C{H}_{2}CH\underset{\_}{H}OLev$$

), 2.71 т (2Н, J_CH2-CH2=6.2, CH₂-Lev), 2.56 т (2Η, CH₂-Lev), 2.15 с (3Н, СН₃-Lev), 1.85 π (2Н, J_CH2-CH2=6.2, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

). ¹³С-ЯМР (CDCl₃): 207.49, 173.12, 167.27 (С=O), 162.98 (С-4), 155.10 (С-2), 146.69 (С-6), 133.50, 133.25, 129.22, 128.18 (Ph), 97.24 (С-5), 95.67 (OCH₂O), 91.60 (С-1′), 85.28 (С-4′), 80.03 (С-2′), 68.60 (С-3′), 65.51 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OLev$$

), 61.87 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 60.86 (С-5′), 38.26 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 30.13 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 29.16 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 28.25 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

).0.75 g (0.98 mmol) of N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -3,5-O- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) β-D-ribofuranosyl} cytosine was treated with 6 ml of a 0.1 Μ solution of tetrabutylammonium fluoride trihydrate in tetrahydrofuran and kept for 10 min at 20 ° C. The reaction mixture was evaporated in vacuo, re-evaporated with chloroform (2 × 10 ml). The residue was chromatographed on a column containing 30 g of silica gel. The substance was eluted with 5% ethanol / dichloromethane. Yield 0.37 g (71%) as a white amorphous foam. R _ƒ 0.20 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 10.20 ct (1H, NHBz), 8.46 d (1H, J _6.5 = 7.5, H-6), 7.93-7.44 m (6H, Bz, H-5), 5.90 d (1H, J _{1′2 ′} = 2.5, H-1 ′), 5.00 d (1H, J = -6.5, $$OC\underset{\_}{H}HO$$

), 4.87 d (1H, $$OCH\underset{\_}{H}O$$

), 4.41 dd (1Н, J _{2′3 ′} = 5.3, Η-2 ′), 4.36 dd (1Н, J _{3′4 ′} = 6.8, Η-3 ′), 4.16-4.09 m (3Н, H-4 ′, $$OC{\underset{\_}{H}}_{2}C{H}_{2}C{H}_{2}OHLev$$

), 4.06 dd (1H, J _5′a5′b = -12.8, J _{5′a4 ′} = 2.2, H-5′a), 3.90 dd (1H, J _5′a5′b = -12.8, J _{5 ′ b4 ′} = 1.9, H-5′b), 3.71-3.62 m (1H, $$OC{H}_{2}C{H}_{2}C\underset{\_}{H}HOLev$$

), 3.64-3.55 m (1H, $$OC{H}_{2}C{H}_{2}CH\underset{\_}{H}OLev$$

), 2.71 t (2H, J _CH2-CH2 = 6.2, CH ₂ -Lev), 2.56 t (2Η, CH ₂ -Lev), 2.15 s (3H, CH ₃ -Lev), 1.85 π (2Н, J _{CH2- CH2} = 6.2, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

) ¹³ C-NMR (CDCl ₃ ): 207.49, 173.12, 167.27 (C = O), 162.98 (C-4), 155.10 (C-2), 146.69 (C-6), 133.50, 133.25, 129.22, 128.18 (Ph ), 97.24 (С-5), 95.67 (OCH ₂ O), 91.60 (С-1 ′), 85.28 (С-4 ′), 80.03 (С-2 ′), 68.60 (С-3 ′), 65.51 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OLev$$

), 61.87 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 60.86 (С-5 ′), 38.26 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 30.13 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 29.16 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 28.25 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

)

c) N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -5-O-monomethoxytrityl-β-D-ribofuranosyl} cytosine.

), 4.93 д (1Н, $$OCH\underset{\_}{H}O$$

), 4.47 дд (1H, J_2′3′=5.0, J_3′4′=8.0, Н-3′), 4.27 д (1Н, Н-2′), 4.20-4.08 м (3Н, Н-4′, Η-5′а, $$OC\underset{\_}{H}HC{H}_{2}C{H}_{2}OLev$$

), 3.80 с (3Н, ОМе), 3.78-3.69 м (1H, $$OCH\underset{\_}{H}C{H}_{2}C{H}_{2}OLev$$

), 3.66-3.52 м (3Н, Η-5′b, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.72-2.70 м (2Н, CH₂-Lev), 2.54-2.51 м (2Н, CH₂-Lev), 2.12 с (3Н, СН₃-Lev), 1.88 п (2Н, J_CH2-CH2=6.2, $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

). ¹³С-ЯМР (CDCl₃): 206.96, 172.84 (С=О), 162.46 (С-4), 158.93 (Ph) 149.01 (С-2), 144.87 (С-6), 144.02, 143.73, 137.02, 135.20, 133.32, 130.46, 129.07, 128.65 128.57, 128.16, 127.93, 127.74, 127.40, 124.16 (Ph), 113.48 (m-PhOMe), 96.80 (С-5), 95.37 (OCH₂O), 89.81 (С-1′), 87.46 (Ph₃C), 83.23 (С-4′), 80.39 (С-2′), 68.13 (С-3′), 65.46 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OLev$$

), 61.63 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 61.35 (С-5′), 55.34 (OMe) 38.07 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 29.89 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 29.06 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 28.06 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

).A solution of 0.37 g (0.69 mmol) of N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -β-D-ribofuranosyl} cytosine and 0.36 mg (1.17 mmol) of monomethoxytrityl chloride in dry pyridine (10 ml) kept for 16 hours at 20 ° C. Methanol (2 ml) was added to the reaction mixture and kept at room temperature for 30 minutes, after which it was evaporated. The residue was diluted with dichloromethane (20 ml) and washed successively with saturated aqueous sodium bicarbonate (20 ml) and water (20 ml). The organic layer was dried over anhydrous sodium sulfate, filtered, evaporated in vacuo. The residue was chromatographed on a column containing 15 g of silica gel. The substance was eluted with a 2% ethanol / dichloromethane system. Yield 0.5 g (90%) as a white foam. R _ƒ 0.41 in the 5% ethanol / dichloromethane system. ¹ H-NMR (CDCl ₃ ): 10.20 ct (1H, NHBz), 8.47 d (1H, J _6.5 = 7.5, H-6), 8.00-7.10 m (17H, Bz, H-5, MMTr), 6.86 d (2H, J = 9.0, m-PhOMe), 5.99 s (1H, H-1 ′), 5.15 d (1H, J = -6.5, $$OC\underset{\_}{H}HO$$

), 4.93 d (1H, $$OCH\underset{\_}{H}O$$

), 4.47 dd (1H, J _{2′3 ′} = 5.0, J _{3′4 ′} = 8.0, H-3 ′), 4.27 d (1H, H-2 ′), 4.20-4.08 m (3H, H-4 ′, Η-5′a, $$OC\underset{\_}{H}HC{H}_{2}C{H}_{2}OLev$$

), 3.80 s (3H, OMe), 3.78-3.69 m (1H, $$OCH\underset{\_}{H}C{H}_{2}C{H}_{2}OLev$$

), 3.66-3.52 m (3Н, Η-5′b, $$OC{H}_{2}C{H}_{2}C{\underset{\_}{H}}_{2}OLev$$

), 2.72-2.70 m (2Н, CH ₂ -Lev), 2.54-2.51 m (2Н, CH ₂ -Lev), 2.12 s (3Н, CH ₃ -Lev), 1.88 p (2Н, J _CH2-CH2 = 6.2 , $$OC{H}_{2}C{\underset{\_}{H}}_{2}C{H}_{2}OLev$$

) ¹³ C-NMR (CDCl ₃ ): 206.96, 172.84 (C = O), 162.46 (C-4), 158.93 (Ph) 149.01 (C-2), 144.87 (C-6), 144.02, 143.73, 137.02, 135.20 , 133.32, 130.46, 129.07, 128.65 128.57, 128.16, 127.93, 127.74, 127.40, 124.16 (Ph), 113.48 (m-PhOMe), 96.80 (C-5), 95.37 (OCH ₂ O), 89.81 (C-1 ′ ), 87.46 (Ph ₃ C), 83.23 (С-4 ′), 80.39 (С-2 ′), 68.13 (С-3 ′), 65.46 ( $$O\underset{\_}{C}{H}_{2}C{H}_{2}C{H}_{2}OLev$$

), 61.63 ( $$OC{H}_{2}C{H}_2\underset{\_}{C}{H}_{2}OLev$$

), 61.35 (С-5 ′), 55.34 (OMe) 38.07 ( $$OCO\underset{\_}{C}{H}_{2}C{H}_{2}COC{H}_3$$

), 29.89 ( $$C{H}_2\underset{\_}{C}{H}_{2}C{H}_2$$

), 06/29 ( $$OCOC{H}_2\underset{\_}{C}{H}_{2}COC{H}_3$$

), 06/28 ( $$OCOC{H}_{2}C{H}_{2}CO\underset{\_}{C}{H}_3$$

)

d) N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -3-O- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) -5-O-monomethoxytrityl-β-D-ribofuranosyl } cytosine (15, B = Cyt ^Bz , R ₁ = MMTr, n = 3).

To a solution of 500 mg (0.62 mmol) of N ^4- Benzoyl-1- {2-O - [(3-levulinyloxypropoxy) methyl] -5-O-monomethoxytrityl-β-D-ribofuranosyl} cytosine in 10 ml of dichloroethane was added diisopropylethylamine ( 0.32 ml, 1.86 mmol) and 2-cyanoethyl-N, N-diisopropylchlorophosphoramidite (0.28 ml, 1.24 mmol). After 2 hours, a 10% aqueous sodium hydrogen carbonate solution (3 ml) was added, and stirred for 10 minutes. A solution of sodium bicarbonate (20 ml) and dichloromethane (20 ml) were added. The organic layer was separated, washed with water (2 × 20 ml), dried with anhydrous sodium sulfate, filtered, evaporated to dryness. The residue was chromatographed on a column containing 15 g of silica gel. The material was eluted with ethyl acetate / hexane / triethylamine (69/29/2). The fractions containing the product were combined, evaporated to dryness in vacuo. Yield 405 mg (65% foam). R _ƒ 0.54 in the dichloromethane / methanol / hexane system (9: 1: 10). ³¹ P-NMR (DMSO-d ₆ ): 152.48, 151.03.

Example 16. Synthesis of N ^6- Benzoyl-9- [2-O- (2,3-di-O-benzoyl-5-O-levulininyl-β-D-ribofuranosyl) -3-O- (2-cyanoethyl-N , N-diisopropylamidophosphinyl) -5-O-dimethoxytrityl-β-D-ribofuranosyl] adenine (16, B = Ade ^Bz ).

The substance was obtained according to published data (SN Mikhailov et al. Chemistry & Biodiversity, 2, 1153-1163, 2005). White powder. R _ƒ 0.43 in the hexane / acetone / triethylamine system (49/49/2). ³¹ P-NMR: 150.94, 150.81.

Example 17. Synthesis of N ^6- Benzoyl-9- {2-deoxy-2- [2- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxyethyl-1-carboxamido] -3-O-tert-butyldimethylsilyl-5-O dimethoxytrityl-β-D-ribofuranosyl} adenine (9, B = Ade ^Bz , R = DMTr, n = 2).

Received from 2′-amino-2′-deoxyadenosine. R _ƒ 0.31 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (CDCl ₃ ): 150.63, 149.29.

Example 18. Synthesis of N ² -Isobutyroyl-9- {2-deoxy-2- [2- (2-cyanoethyl-N, N-diisopropylamidophosphinyl) hydroxyethyl-1-carboxamido] -3-O-tert-butyldimethylsilyl-5-O -dimethoxytrityl-β-D-ribofuranosyl} guanine (9, B = Gua ^iBu , R = DMTr, n = 2) Received from 2′-deoxy-2′-aminoguanosine. R _ƒ 0.30 in the system ethyl acetate / hexane / triethylamine (69/29/2). ³¹ P-NMR (CDCl ₃ ): 150.40, 149.10.

Example 19. Synthesis of an oligomeric mimetic VAP C ₇ H ₁₆ NO ₂ - (5′pAraAdo5 ′) ₁₁ (Fig. 11)

Solid-phase synthesis of the oligomer was carried out on an ABI 3400 automatic synthesizer (Applied Biosystems, USA) using glass with a controlled pore size (500 Å) as a solid support with covalently sewn protected C-7 aminolinker. The synthesis temperature is 20 ° C. The synthesis scale was 0.2 μmol. A standard synthesis program was used. The cycle of joining one monomer unit consisted of 4 stages (Fig. 11):

1. Condensation (addition of an amidophosphite block dissolved in acetonitrile or dichloromethane at a concentration of 0.5-0.05 M in the presence of 1H-tetrazole) was carried out using amidophosphite disaccharide nucleoside 3 at 20 ° C (stage i). The condensation time was set equal to 300 seconds.

2. Capping (stage ii - acylation of unreacted hydroxyl groups with a mixture of propionic anhydride and pyridine in a ratio of 1: 1 to 1: 3 in tetrahydrofuran) - 20-60 sec at 20 ° C.

4. Oxidation of 0.05-0.1 Μ with iodine solution in a mixture of tetrahydrofuran-pyridine-water (stage iii - conversion of internucleotide phosphite to phosphotriether, X = O) -15-20 sec at 20 ° C. Oligonucleotide phosphordithioates (stage iii, X = S) are prepared by the reaction of internucleotide phosphites with 3 - ((dimethylaminomethylene) amino) -3H-1,2,4-dithiazole-3-thione in a tetrahydrofuran-pyridine mixture (concentration of sulfonating reagent 0.05-0.1 M, the reaction time is 360-600 sec at 20 ° C).

5. Release (stage iv — removal of the trityl protecting group from the 5 'hydroxyl group of the oligonucleotide chain with a 2-5% solution of di- or trichloroacetic acid) - 60 sec at 20 ° C. The removal of monomethoxytrityl protection (MMTr) before the condensation step was carried out manually using the function '' Hold ''.

Sequential attachment of the links was carried out repeating steps i-iv (FIG. 11). The last MMTr group was not removed. Removal of acetyl protective groups and cleavage of the oligomer from the support was carried out under standard conditions (conc. Aqueous ammonia, 6 hours, 55 ° C, stage v, AraAdo - designation of a disaccharide nucleoside, Tfa-trifluoroacetyl, CEt - β-cyanoethyl, pr (abbr. Protecting groups) -protected groups to be deleted). A partially unblocked oligomer was isolated by reverse phase HPLC (Hypersil C18, 5mkm, 4.6 × 250 mm) at 0.05 Μ TEAA and an acetonitrile gradient (10-50% in 30 min). After the MMTr of the protective group was removed with 80% aqueous acetic acid for 30 min at room temperature (step vi), the oligomer was re-purified in the indicated buffer and a gradient of acetonitrile from 0 to 25% (30 min). The yield of oligomer after isolation was 20 p.u. (260 nm).

Example 20. Synthesis of oligomeric mimetic PAR Eosin-C ₇ H ₁₆ NO ₂ - (5′pAraAdo5 ′) ₁₁ .

The oligomer C ₇ H ₁₆ NO ₂ - (5′′pAraAdo5 ′) ₁₁ (2 pu) was dissolved in 50 μl of 0.1 Μ Na-bicarbonate buffer (pH 9) and 50 μl of a solution of eosin succinimide ether (0 ° C) was added at 0 ° C ( 1 mg) in anhydrous DMF. The mixture was left for 16 hours at 4 ° C. Then, the solution volume was adjusted to 1 ml and the unreacted dye was extracted with butanol to a final volume of the aqueous phase of 100 μl. The oligomer was planted with 2% LiClO ₄ in acetone. The conjugate was isolated by reverse phase HPLC (Hypersil C18, 5mkm, 4.6 × 250 mm) at 0.05 Μ TEAA and an acetonitrile gradient (10-50% in 30 min). The yield of oligomer after isolation was 1 p.u. (260 nm).

Example 21. The synthesis of a branched oligomeric mimetic PAR 3′-dAdo (5′′pAraAdo5 ′) ₅ -X - [(5′′pAraAdo5 ′) ₅ ] ₂ , X = asymmetric splitter).

The oligomer synthesis was carried out according to the standard scheme as described in example 19, with minor changes. Glass with a controlled pore size (1000 Å) and covalently attached protected deoxyadenosine was used as a solid support. The condensation time of the amidophosphite of an asymmetric splitter was set to 15 minutes. Upon completion of the synthesis of the main branch of the oligomer, the MMTr protecting group was removed and the free hydroxyl group was acylated with a caper mixture for 30 min at 20 ° C. Then, the Fmoc protection of the 1M DBU splitter in anhydrous MeCN was removed 5 min at room temperature. Then, the oligomer side chain was synthesized. The yield of oligomer after isolation was 10 p.u. (260 nm).

Example 22. Activation of PARP-1 with PAR mimetics.

To 40 μl of a solution of NAD + in a PARP buffer (50 mM Tris-HCl (pH 8.0), 2 mM MgCl ₂ ) was added 20 μl of a mimetic PAR (1-10 μM) or activated DNA, 20 μl of water or an inhibitor solution (when using inhibitor as a control), 1 μl PARP-1 in 19 μl PARP buffer and incubated at 20 ° C for 30 min. After incubation, 40 μl of 2M KOH and acetophenone solutions (20% in alcohol) were added, shaken, and incubated for 10 min at 4 ° C. Then 180 μl of 88% formic acid solution was added, vortexed and incubated for 10 min at 110 ° C. The resulting mixture was diluted to 1500 μl with PARP buffer and the fluorescence of the solution was measured at 440 nm. Fluorescence of the solution in the absence of a PARP-1 inhibitor corresponded to the minimum content of residual NAD + and maximum enzyme activity. Fluorescence of a solution containing no enzyme, inhibitor, and activated DNA corresponds to the maximum NAD + content and zero enzyme activity. For calibration purposes, we measured the fluorescence of solutions with NAD + content 2, 10, and 100 times lower than indicated above. The specific activity of PARP-1 was at least 600 units / mg. One unit of PARP-1 is the amount of enzyme required for the synthesis of 1 nmol poly (ADP-ribose) per minute at 25 ° C and pH 7.5. The concentration of the enzyme was 1 mg / ml in accordance with the certificate of production.

Claims (2)

1. PAI mimetics of the general formula

where Y is a nucleoside residue such as deoxyadenosine, deoxyguanosine, adenosine, guanosine, cytidine, uridine; an amino derivative of a linear or branched aliphatic compound C7, such as 6-amino (2-hydroxymethyl) hexan-1-yl; a fluorescent dye, such as fluorescein or eosin,
where Z is a nucleoside residue such as deoxyadenosine, deoxyguanosine, adenosine, guanosine, cytidine, uridine, optionally with a 5′-hydroxyl group attached to the residue of monomethoxytrityl or dimethoxytrityl,
(k + 1) m = 1-200; X represents O or S;
R ₁ and R ₂ are a disaccharide nucleoside residue which is a nucleoside with an additional carbohydrate residue attached to the pentafuranose nucleoside cycle of an O-glycosidic bond, or a residue of the formulas

n is 2-6; or

n is 2-6; or

n = 1-4 or - ((CH ₂ ) _n O) _m - (P = X (OH)) ON-, where n = 2-6, m = 1-6, or

where N = residue of a nucleoside such as deoxyadenosine, deoxyguanosine, adenosine, guanosine, cytidine, uridine,
R ₃ is a splitter

where N′ is the residue of a disaccharide nucleoside; n is the number of monomer units with a degree of polymerization of up to 100 monomer units, or the remainder of one of the formulas

n is 2-6; or

n is 2-6; or

n = 1-4
where B = adenin-9-yl, B = uracil-1-yl, B = cytosin-1-yl, B = guanine-9-yl. 2. A method for producing PAR mimetics described in Clause 1 on an insoluble silicate, polystyrene or polyacrylate carrier at a temperature of 20-40 ° C with sequential addition of amidophosphite monomer blocks 1-16 in the course of synthesis to obtain the polymer of the required length:

where R = MMTr, DMTr, B = N ⁶ -benzoyladenin-9-yl, N ² -isobutyroylguanin-9-yl, N ⁴ -benzoylcytosin-1-yl, uracil-1-yl, n = 2-6 for amidophosphites 7 -8, n = 1-4 for amidophosphite 9, in manual or automatic mode, where each reaction cycle includes: 1) the release of a 2-5% solution of di- or trichloroacetic acid in dichloromethane, 2) a condensation step using 0, 05-0.5 M solution of amidophosphite in acetonitrile or dichloromethane; 3) acylation of unreacted hydroxyl groups with a mixture of propionic anhydride and pyridine in a ratio of 1: 1 to 1: 3 in tetrahydrofuran; 4) the oxidation of the internucleotide phosphite obtained in the previous stage to phosphotriether with a 0.05-0.1 M solution of iodine in a mixture of tetrahydrofuran-pyridine-water; followed by the release of the resulting mimetic under standard conditions with aqueous ammonia and purification of the product using reverse phase HPLC.

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