RU2518369C1 - Alkyl [2-(2-{5-[4-(4-{2-[1-(2-methoxycarbonylamino-acetyl)-pyrrolidin-2-yl]-3h-imidazol-4-yl}-phenyl)-buta-1,3-diinyl]-1h-imidazol-2-yl}-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamate, pharmaceutical composition, medication, method of treating viral diseases - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to novel alkyl [2-(2-{5-[4-(4-{2-[1-(2-methoxycarbonylamino-acetyl)-pyrrolidin-2-yl]-3H-imidazol-4-yl}-phenyl)-buta-1,3-diinyl]-1H-imidazol-2-yl}-pyrrolidin-1-yl)-2-oxo-ethyl]-carbamates or their naphthalene-1,5-disulfonates, which possess properties of NS5A protein inhibitor and can be used for prevention and treatment of viral diseases, caused by viruses of hepatitis C (HCV) and hepatitis GBV-C. In claimed invention compounds, corresponding to general formula 1

R¹, R², R³ and R⁴ independently on each other stand for C₁-C₃ alkyl; R⁵ and R⁶ independently on each other stand for C₁-C₃alkyloxymethyl, or R³, and R⁵, and R⁴, and R⁶, together with carbon atoms, which they are bound with, independently on each other form tetrahydrofurane cycle.

EFFECT: improved propertied of compounds.

7 cl, 2 tbl, 7 ex

Description

The present invention relates to NS5A inhibitors, to a new pharmaceutical composition, an antiviral drug, a method for the prevention and treatment of viral diseases, especially caused by hepatitis C viruses (HCV).

Viral infections can cause many diseases, which poses a serious threat to human health and life. Over the past 20 years, at least 30 completely new pathogens of infectious diseases have been discovered: AIDS, viral hepatitis, acute and chronic diarrhea, hemorrhagic fevers (Ebola, Venezuelan, Brazilian, Rift Valley) [a) Lednicky J.A., Rayner J.O. Uncommon respiratory pathogens. Curr. Opin. Pulm. Med. 2006, 12 (3), 235-239; b) Hayden F.G. Respiratory viral threats. Curr. Opin. Infect. Dis. 2006, 19 (2), 169-178]. In particular, the possibility of human disease with the so-called bird flu [a) Liu J.P. is of particular concern. Avian influenza-a pandemic waiting to happen, J. Microbiol. Immunol. Infect. 2006, 39 (1), 4-10; b) Henter J.I .; Chow C. B .; Leung C.W, Lau Y.L. Cytotoxic therapy for severe avian influenza A (H5N1) infection. Lancet. 2006 367 (9513), 870-873. Review]. According to statistics, 60-65% of epidemic infections have a viral etiology. Due to the complexity of the interaction in the triad “virus-host organism-drug”, most modern antiviral drugs in the course of therapy exhibit side effects and form resistant viral strains [Jain R., Clark N.M., Diaz-Linares M., Grim S.A. Limitations of current antiretroviral agents and opportunities for development. Curr. Pharm. Des. 2006, 12 (9), 1065-1074]. Currently, the number of antiviral drugs that can be used in clinical practice is extremely limited - only 43 low molecular weight substances [http://integrity.prous.com/integrity], which far from meets the needs of the prevention and treatment of viral diseases. In addition, there are a significant number of viral infections that cause diseases for which no chemotherapeutic agents currently exist. This applies, for example, to diseases caused by papilloma viruses, adenoviruses, herpes-6, smallpox, SARS syndrome, hemorrhagic fevers, West Nile fever, bird flu, etc. [De Clercq E. Recent highlights in the development of new antiviral drugs. Curr Opin Microbiol. 2005, 8 (5), 552-560].

Hepatitis C virus belongs to flaviviruses (genus Flaviviridae), along with other important human pathogens, such as yellow fever virus, West Nile virus, Dengue virus and hepatitis GBV-C virus. Flaviviruses have similar genome organization, including the presence of a gene encoding an NS5A non-structural protein. NS5A plays an important role in the replication of the viral RNA genome, being a structural component of the viral replication complex. Since this protein is currently validated in clinical trials as a target for the development of drugs against chronic hepatitis C, NS5A seems to be a promising target for other clinically important flaviviruses listed above.

Thus, the creation of new antiflavovirus drugs, especially those with high activity and low toxicity, is of great importance.

There are publications in the patent literature on various derivatives of 2-pyrrolidin-2-yl-1H-imidazoles, which are ligands of the non-structural protein NS5A and suppress the hepatitis C virus (HCV) [WO 2008021927 A2, WO 2009020825 A1, WO 2009020828 A1, WO 2010065668 A1, WO 2010065681 A1, WO 2010096302 A1, WO 2010096462 A1, WO 2010096777 A1, WO 2010111534 A1, WO 2010111673 A1, WO 2010117635 A1, WO 2010117977 A1], including the patent RU 2452735 C1, 2012, which describes methyl [(S) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) -pyrrolidine -2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] car Amat (A) and the dihydrochloride · 2HCl.

However, due to the bulkiness of the molecules, the purification of this type of compounds both in the form of bases A and in the form of A · 2CHl hydrochloride is very difficult. This makes them difficult to test and use as drug candidates.

"Alkyl" means an aliphatic hydrocarbon linear or branched group with 1-5 carbon atoms in the chain. Branched means that the alkyl chain has one or more substituents, for example C ₁ -C ₃ alkyloxymethyl.

“Medicinal substance” (drug substance, drug substance, drug-substance, active ingredient) means a physiologically active substance of synthetic or other (biotechnological, plant, animal, microbial and other) origin, having pharmacological activity and being the active principle of the pharmaceutical composition used for production and manufacture of a medicinal product (product).

“Medicinal product (preparation)” - a substance (or a mixture of substances in the form of a pharmaceutical composition) in the form of tablets, capsules, injections, ointments and other formulations intended to restore, correct or alter physiological functions in humans and animals, as well as for treatment and disease prevention, diagnosis, anesthesia, contraception, cosmetology and more.

A “therapeutic cocktail” is a simultaneously administered combination of two or more drugs with a different mechanism of pharmacological action and aimed at different biological targets involved in the pathogenesis of the disease.

"Pharmaceutical composition" means a composition comprising at least one of the claimed compounds and at least one of the components selected from the group consisting of pharmaceutically acceptable and pharmacologically compatible excipients, solvents, diluents, carriers, excipients, distributing and perceiving means of delivery, such as preservatives, stabilizers, fillers, grinders, moisturizers, emulsifiers, suspending agents, thickeners, sweeteners, perfumes, flavors, and antibacterial agents, fungicides, lubricants, prolonged delivery regulators, the choice and ratio of which depends on the nature and method of administration and dosage. Examples of suspending agents are ethoxylated isostearyl alcohol, polyoxyethylene, sorbitol and sorbitol ether, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, as well as mixtures of these substances. Protection against the action of microorganisms can be achieved using a variety of antibacterial and antifungal agents, for example, such as parabens, chlorobutanol, sorbic acid and the like. The composition may also include isotonic agents, for example, sugars, sodium chloride and the like. The prolonged action of the composition can be achieved using agents that slow down the absorption of the active principle, for example aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery vehicles are water, ethanol, polyalcohols, and also mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Examples of excipients are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate and the like. Examples of grinders and distributors are starch, alginic acid and its salts, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol. The pharmaceutical composition for oral, sublingual, transdermal, intramuscular, intravenous, subcutaneous, local or rectal administration of the active principle, alone or in combination with another active principle, can be administered to animals and humans in a standard administration form, in the form of a mixture with traditional pharmaceutical carriers. Suitable unit dosage forms include oral forms such as tablets, gelatine capsules, pills, powders, granules, chewing gums and oral solutions or suspensions, sublingual and buccal administration forms, aerosols, implants, local, transdermal, subcutaneous, intramuscular, intravenous, intranasal or intraocular administration forms and rectal administration forms.

The authors found that previously unknown steroisomers of alkyl [2- (2- {5- [4- (4- {2- [1- (2-methoxycarbonylamino-acetyl) pyrrolidin-2-yl] -3H-imidazole-4- yl} -phenyl) -buta-1,3-diinyl] -1H-imidazol-2-yl} -pyrrolidin-1-yl) -2-oxoethyl] carbamates of the general formula 1, possibly in crystalline or polycrystalline form, exhibit high activity against hepatitis C virus and hepatitis GBV-C virus,

where R ¹ , R ² , R ³ and R ⁴ independently from each other represent C ₁ -C ₃ alkyl;

R ⁵ and R ⁶ independently of one another are C ₁ -C ₃ alkyloxymethyl or R ³ _, and R ⁵ _, and R ⁴ _, and R ^6, together with the carbon atoms to which they are bonded, independently form a tetrahydrofuran ring.

The authors also found that previously unknown stereoisomers of alkyl [2- (2- {5- [4- (4- {2- [1- (2-methoxycarbonylamino-acetyl) pyrrolidin-2-yl] -3H-imidazole-4 -yl} -phenyl) -buta-1,3-diinyl] -1H-imidazol-2-yl} -pyrrolidin-1-yl) -2-oxoethyl] carbamate naphthalene-1,5-disulfonates of the general formula 1.1 , possibly in crystalline or polycrystalline form, exhibit high activity against hepatitis C virus and GBV-C hepatitis virus,

Where

R ¹ , R ² , R ³ and R ⁴ independently of one another are C ₁ -C ₃ alkyl;

R ⁵ and R ⁶ independently of one another are C ₁ -C ₃ alkyl or C ₁ -C ₃ alkyloxymethyl, or

R ³ _, and R ⁵ _, and R ⁴ _, and R ⁶ together with the carbon atoms to which they are attached independently form a tetrahydrofuran ring,

excluding alkyl [(S) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) -pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diinyl) phenyl] -1H-imidazol-2-yl} -pyrrolidin-1-carbonyl) -2-methyl-propyl ] carbamate naphthalene-1,5-disulfonate.

Preferred stereoisomers are alkyl [2- (2- {5- [4- (4- {2- [1- (2-methoxycarbonylamino-acetyl) pyrrolidin-2-yl] -3H-imidazol-4-yl} phenyl) -buta-1,3-diinyl] -1H-imidazol-2-yl} -pyrrolidin-1-yl) -2-oxoethyl] carbamate naphthalene-1,5-disulfonates are:

methyl [(R) -1 - ((R) -2- {5- [4- (4- {2 - [(R) -1 - ((R) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate 1.1.1;

methyl [(S) -1 - ((R) -2- {5- [4- (4- {2 - [(R) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate 1.1.2;

methyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate 1.1.3;

methyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate 1.1.4;

methyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((R) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate 1.1.5;

isopropyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((R) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate 1.1.6;

methyl [2 - ((S) -2- {5- [4- (4- {2- (S) -1- (2-methoxycarbonylamine-2-methyl-propionyl) pyrrolidin-2-yl] -3H- imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-yl) -1,1-dimethyl-2-oxoethyl] carbamate naphthalene -1.5-disulfonate 1.1.7;

methyl N- [1 - ({((2S) 2- [5- (4- {4- [2 - ((2S) -1- {2 - [(methoxycarbonyl) amino] -2-methyl-3-methoxypropioyl} tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrol-1-yl} carbonyl) - 1-methyl-2-methoxyethyl] carbamate naphthalene-1,5-disulfonate 1.1.8;

methyl N- [3 - ({(2S) -2- [5- (4- {4- [2 - ((2S) -1- {3 - [(methoxycarbonyl) amino] -tetrahydrofuran-3-ylcarbonyl} tetrahadro -1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrol-1-yl} carbonyl) -tetrahydrofuran -3-yl] carbamate naphthalene-1,5-disulfonate 1.1.9;

methyl N - [(3S) -3 - ({(2S) -2- [5- (4- {4- [2 - ((2S) -1 - {(3S) -3 - [(methoxycarbonyl) amino] -tetrahydrofuran-3-ylcarbonyl} tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrole- 1-yl} carbonyl) -tetrahydrofuran-3-yl] carbamate naphthalene-1,5-disulfonate 1.1.10.

In accordance with this invention, the salts of naphthalene-1.5-disulfonates of the steroisomers 1.1.1-1.1.10 are prepared by reacting the corresponding bases of formula 1 with naphthalene-1,5-disulfonic acid in methanol, ethanol or isopropanol.

The salts obtained according to this invention are distinguished by the high purity required for use in pharmacopoeia and are well stored at low temperatures (from 4.7 ° C to -23 ° C).

In turn, the bases of formula 1 were obtained using well-known chemical reactions and the corresponding optically active commercial reagents according to known methods described in patent RU 2452735 C1 (2012).

The structure of the new compounds and their purity are confirmed by analytical chromatography, LC MS and NMR spectroscopy. The compounds are named using the ChemDrow program (Chembridge Soft Inc.).

The subject of this invention is NS5A protein inhibitors, which are compounds of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10, possibly in crystalline or polycrystalline form.

The subject of this invention is a pharmaceutical composition for the treatment and prevention of diseases caused by hepatitis C virus and hepatitis GBV-C virus, containing, as an NS5A protein inhibitor, a compound of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10 in therapeutically effective amount.

Pharmaceutical compositions may include pharmaceutically acceptable excipients. By pharmaceutically acceptable excipients are meant diluents, excipients and / or carriers used in the pharmaceutical field. The pharmaceutical composition, along with the compound of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10, or its crystalline or polycrystalline form, the present invention may include other active ingredients, provided that they do not cause undesirable effects e.g. allergic reactions.

If it is necessary to use the pharmaceutical compositions of the present invention in clinical practice, they can be mixed for the manufacture of various forms, and they may include traditional pharmaceutical carriers, for example, oral forms (such as tablets, gelatine capsules, pills, solutions or suspensions); injection forms (such as solutions or suspensions for injection, or dry powder for injection, which requires only the addition of water for injection before use); local forms (such as ointments or solutions).

The carriers used in the pharmaceutical compositions of the present invention are carriers that are used in the pharmaceutical field to obtain common forms, including: in oral forms, binders, lubricants, disintegrants, solvents, diluents, stabilizers, suspending agents, colorless are used agents, flavoring agents of taste; antiseptic agents, solubilizers, stabilizers are used in injection forms; in local forms, bases, diluents, lubricants, antiseptic agents are used.

A pharmaceutical composition is prepared by mixing at least one compound of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10, or its crystalline or polycrystalline form with an inert filler and / or solvent.

The clinical dosage of a pharmaceutical composition containing, as an active component, a compound of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10 in patients can be adjusted depending on the therapeutic efficacy and bioavailability of the active ingredients in the body, their metabolic rate and excretion from the body, as well as depending on the age, gender and stage of the patient’s disease, while the daily dose in adults is usually 10 ~ 500 mg. Therefore, during the preparation of the pharmaceutical compositions of the present invention in the form of dosage units, the above effective dosage must be taken into account, with each dosage unit of the preparation containing 10 ~ 500 mg of the compound of general formula 1, or 1.1, or of formulas 1.1.1-1.1.10. In accordance with the instructions of a doctor or pharmacist, these drugs can be taken several times during certain periods of time (preferably from one to six times).

The subject of this invention is a medicament in the form of tablets, capsules or injections in a pharmaceutically acceptable package for the treatment and prevention of diseases caused by hepatitis C virus and GBV-C hepatitis virus, containing at least one compound of general formula 1 as an inhibitor of the NS5A protein or 1.1, or formulas 1.1.1-1.1.10, or a new pharmaceutical composition in a therapeutically effective amount.

The subject of this invention is a therapeutic cocktail for the prevention and treatment of diseases caused by hepatitis C virus and hepatitis GBV-C virus, comprising at least one compound of general formula 1, or 1.1, or formulas 1.1.1-1.1.10, or a new a pharmaceutical composition, or a new drug in a therapeutically effective amount.

Therapeutic cocktails for the prevention and treatment of the above flavavirus diseases, including hepatitis C and hepatitis GBV-C, along with the drugs of this invention, may include: inosine-5-monophosphate dehydrogenase inhibitors, for example Ribavirin (allowed) and Ribamidine; hepatitis C protease NS3 inhibitors, for example Telaprevir, Siluprevir and SCH-503034; NS5B RNA polymerase inhibitors, for example XTL-2125; alpha glucosidase inhibitors, for example, amino carbohydrate Selgozivir; as well as TLR receptor agonists, hepatoprotectors, cyclosporins, various proteins (e.g. interferons), antibodies, vaccines, etc.

For combination therapy, any classes of agents may be useful when combined with the bis-azoles of the present invention, and may include, for example, nucleoside and non-nucleoside inhibitors of HCV polymerase, protease inhibitors, helicase inhibitors, NS4B inhibitors and medical agents that functionally inhibit the internal ribosome entry site (IRES) and other medicines that inhibit the attachment of HCV cells or virus entry, HCV RNA transcription, HCV RNA transcription, replication or HCV maturation, focusing ix or attenuated virus. Specific compounds in these classes and useful in this invention include, but are not limited to, macrocyclic, heterocyclic, and linear HCV HCV protease inhibitors such as telaprevir (VX-950), boceprevir (SCH-503034), narlaprevir (SCH-900518), ITMN -191 (R-7227), TMC-435350 (aka TMC-435), MK-7009, BI-201335, BI-2061 (ciluprevir), BMS-650032, ACH-1625, ACH-1095 (HCV NS4A inhibitor of concomitant factor proteases) VX-500, VX-813, PHX-1766, PHX2054, IDX-136, IDX-316, ABT-450 EP-013420 (and related) and VBY-376; nucleoside HCV polymerase inhibitors (replicases) useful in the invention include, but are not limited to, the following: R7128, PSI-7851, IDX-184, IDX-102, R1479, UNX-08189, PSI-6130, PSI-938 and PSI -879 and various other nucleosides and nucleotide analogs and HCV inhibitors, including (but not limited to) derived from 2′-C-methyl modified nucleosides and nucleotides; and 7′-dease modified nucleosides and nucleotides. Inhibitors of non-nucleoside HCV polymerase (replicase) useful in this invention include, but are not limited to: HCV-796, HCV-371, VCH-759, VCH-916, VCH-222, ANA-598, MK-3281, ABT-333 , АВТ-072, PF-00868554, BI-207127, GS-9190, А-837093, JKT-109, GL-59728 and GL-60667.

In addition, the NS5A inhibitors of the present invention can be used in combination with cyclophillin and immunophillin antagonists (e.g., without limitation of the DEBIO compound, NM-811, as well as cyclosporin and its derivatives), kinase inhibitors, heat shock protein inhibitors (e.g. HSP90, HSP70 ), other immunomodulatory agents, which may include, without limitation, interferons (alpha, beta, omega, gamma, lambda or synthetic), such as Intron A ™, Roferon-A ™, Canferon-A300 ™, Advaferon ™, Infergen ™, Humoferon ™, Sumiferon MP ™, Alfaferon ™, IFN-β ™, Feron ™, and the like, interferon Connections, polyethylene glycol derivatized (pegylated), such as PEG interferon-α-2a (Pegasys ™), PEG interferon-α-2b (PEGIntron ™), pegylated IFN-α-con 1 and the like; prolonged formulas and derivatives of interferon compounds, such as albumin-fused interferon, Albuferon ™, Locteron ™, and the like; interferons with various types of controlled delivery (for example, ITCA-638, omega-interferon delivered by DUROS subcutaneous delivery system); compounds that stimulate the synthesis of interferon in cells, such as resiquimod and the like; interleukins; compounds that enhance the development of type 1 helper T cell response, such as SCV-07 and the like; TOLL - like receptor agonists, such as CpG-10101 (action), isotorabine, ANA773 and the like; thymosin α-1, ANA-245 and ANA-246, histamine dihydrochloride, propagermanium; tetrachlorodecaoxide; ampligen; IMP-321; KRN-7000; antibodies, such as: civacir, XTL-6865 and the like; and prophylactic and therapeutic vaccines such as Inno Vac, HCV E1E2 / MF59 and the like. In addition, any of the methods described above, including the administration of an NS5A inhibitor, a type I interferon receptor agonist (e.g. IFN-α), and a type II interferon receptor agonist (e.g. IFN-γ), can be enhanced by the administration of an effective amount of a TNF-α antagonist. A typical non-limiting TNF-α antagonist that is suitable for use in such combination therapy is ENBREL ™ and HUMIRA ™.

In addition, the NS5A inhibitors of the present invention can be used in combination with antiprotozoans, and other antiviruses are considered effective in the treatment of HCV infection, such as the prodrug nitazoxanide. Nitazoxanide can be used as an agent in combination with the compounds disclosed in this invention, as well as in combination with other agents useful in the treatment of HCV infection, such as peginterferon alfa-2a and ribavarin (e.g. Rossignol, JF and Keeffe, EB, Future Microbiol. 3: 539-545, 2008).

The NS5A inhibitors of the present invention can also be used with alternative forms of interferons and pegylated interferons, ribavirin or its analogues (e.g. Tarabavarin, levovirion), microRNA, low-harm RNA compounds (e.g. SIRPLEX-140-N and the like), nucleotide or nucleoside analogs, immunoglobulins, hepatoprotectors, anti-inflammatory agents and other NS5A inhibitors. Inhibitors of other targets in the HCV life cycle include NS3 helicase inhibitors; co-factor NS4A inhibitors; antisense oligonucleotide inhibitors such as ISIS-14803, AVI-4065 and the like; vector-encrypted short hairpin RNA (shRNA); HCV specific ribozymes such as heptazyme, RPI, 139199 and the like; entry inhibitors such as HepX-C, HuMax-Heps and the like; alpha glucosidase inhibitors such as celgosivir, UT-231B and the like; KRE-02003002 and BIVN 401 and IMPDH inhibitors. Other representative compounds — HCV inhibitors — include those disclosed in well-known scientific and patent publications.

Additionally, combinations, for example ribavirin and interferon, can be administered as combination therapy with at least one of the compounds of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10.

The present invention is not limited to the above classes or compounds, and contemplates known and novel compounds and combinations of biologically active agents. It is understood that the combination therapy of the present invention includes any chemical compatible combinations of the bis-azoles of this patentable group with other compounds of the patentable group or other compounds outside the patentable group, and the combination does not exclude the antiviral activity of the compounds of this patentable group or the antiviral activity of the pharmaceutical composition itself.

Combination therapy may be sequential, i.e. treatment first with one agent and then with another (for example, when each treatment step involves a different compound of the present invention or when one treatment step includes a compound of the present invention and the other involves one or more biologically active agents), or there may be a treatment with both agents simultaneously . Sequential therapy may include a significant amount of time after completion of the first therapy and before the start of the second therapy. Treatment with both agents at the same time can be carried out in a single daily dose or in different doses. Combination therapy does not need to be limited to two agents and may include three or more agents. Doses for simultaneous and sequential combination therapy will depend on absorption, distribution, metabolic rates and excretion of the components of combination therapy, as well as other factors well known to the specialist. The size of the dose will also vary depending on the severity of the condition that needs to be alleviated. It should be understood that for each particular subject, the specific dosage regimen and schedule can be adjusted in time in accordance with the needs of the individual and the professional judgment of the person who treats or monitors the treatment using the combination therapy method.

The subject of this invention is a method for the prevention and treatment of diseases caused by hepatitis C virus and hepatitis GBV-C virus, by administering a therapeutically effective amount of at least one compound of general formula 1, or general formula 1.1, or formulas 1.1.1-1.1.10, or a new pharmaceutical composition, or a new drug, or a therapeutic shake.

The following examples demonstrate, but are not limited to, the present invention.

Example 1. General method for the production of alkyl [2- (2- {5- [4- (4- {2- [1- (2-methoxycarbonylamino-acetyl) -pyrrolidin-2-yl] -3H-imidazole-4- sterioisomers yl} -phenyl) -buta-1,3-diinyl] -1H-imidazol-2-yl} -pyrrolidin-1-yl) -2-oxoethyl] carbamates of the general formula 1 [1 (1) -1 ( 3)]

To a solution of 8.15 g (42 mmol) of 1,4-bis- (trimethylsilyl) -1,3-butadiene in 50 ml of dry ether under argon, 31 ml (46.5 mmol) of a 1.5 M solution of MeLi × LiBr in broadcast. The mixture was stirred for 6 hours at room temperature. 50 ml of a saturated solution of NH ₄ Cl were slowly added to the mixture, extracted with pentane, dried over Na ₂ SO ₄ and the solvents were removed in a weak vacuum. The liquid residue was dissolved in 70 ml of THF, 20 ml of triethylamine, 8.8 g (20 mmol) of compound 1 (1) .1, 458 mg (0.5 mmol) Pd ₂ (dba) ₃ , 524 mg (2 mmol) were added. triphenylphosphine, 190 mg (1 mmol) CuI, and the mixture was stirred under argon for 12 hours at 40 ° C. The mixture was filtered through celite, applied to silica gel and compound 1 (1) .2 was isolated by flash chromatography (eluent CHCl ₃ : ErOAc = 10: 1). Yield 7.56 g (87%). LCMS (M + H) ⁺ 434.

To a solution of 7.36 g (17 mmol) of compound 1 (1) .2 in 120 ml of THF and 120 ml of methanol was added 7.04 g (51 mmol) of potash and the mixture was stirred for 2 hours. The solvents were removed in vacuo and the residue was treated with 150 ml. THF and filtered. To the resulting solution of compound 1 (1) .3, 5.56 g (15.3 mmol) of compound 13-1 (5-iodo-2 - [(28) -1-vos-pyrrolidin-2-yl] -1H- imidazole), 366 mg (0.4 mmol) of Pd ₂ (dba) ₃ , 630 mg (2.4 mmol) of triphenylphosphine, 152 mg (0.8 mmol) of CuI, and the mixture was stirred under argon for 12 hours at 40 ° C. The mixture was filtered through celite, applied to silica gel and compound 1 (1) .4 was separated from the main part of the impurities by flash chromatography (eluent CHCl ₃ : MeOH = 80: 1). After evaporation of the solvent, the residue is treated with 60 ml of acetonitrile, kept in an ultrasound bath until crystallization begins and left for 3 hours. The precipitate formed is filtered off, washed with acetonitrile, ether and dried in vacuo. Yield 5.47 g (54%). LCMS (M + H) ⁺ 597.

15 ml of a 3 M solution of HCl in dioxane were added to compound 1 (1) .4 (0.695 g) and stirred for 12 hours. The mixture was evaporated in vacuo to give compound 1 (1) .5 in 55% yield. LCMS (ESI): LCMS (M + H) ⁺ 397. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 2.02 (br.s., 8H), 2.99 (m, 4H), 3.40 (br.s. , 2H), 6.22 (m, 4H), 6.75 (s, 1H), 7.22 (s, 1H), 7.77 (m, 2H), 7.83 (d, 2H).

A mixture of 50 mg (0.244 mmol, 2.1 equiv.) 4-methoxy-N-methoxycarbonyl-L-valine, 40 mg (0.295 mmol, 2.5 equiv.) 1-hydroxybenzotriazole and 53 mg (0.277 mmol, 2.35 equiv.) N- ( 3-dimethylaminopropyl) -N′-ethylcarbodiimide hydrochloride in 1 ml of acetonitrile was stirred for 1 h, 64 mg (0.118 mmol, 1 equiv.) Of compound 1 (1) were added. 5 and 82 μl (61 mg, 0.472 mmol, 4 equiv.) diisopropylethylamine. The reaction mass is stirred for 12 hours at room temperature. The completeness of the reaction is monitored by LCMS. After completion of the reaction, the solvent was completely distilled off on a rotary evaporator, the residue was dissolved in dichloromethane. The extract was washed with 10% sodium carbonate solution, dried over Na ₂ SO ₄ and evaporated on a rotary evaporator. Further purification is carried out by HPLC. Methyl N- [1 - ({((2S) -2- [5- (4- {4- [2 - ((2S) -1- {2 - [(methoxypropioyl) amino])] -2-methyl-3- methoxypropioyl} tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-buta-diinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrol-1-yl } carbonyl) -1-methyl-2-methoxyethyl] carbamate 1 (1) in 71% yield. LCMS (M + H) ⁺ 744, ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.56 (s, 3H), 1.65 (s, 3H), 1.65 (m, 2H), 1.85 (m, 2H), 2.05 (m, 4H), 3.43 (s, 3H), 3.51 (s, 3H), 3.61 (s, 6H), 3.66 (m, 2H), 3.75 (br.s, 2H), 3.89 (m, 2H) 4.11 (br.s, 2H), 5.00 (m, 2H), 6.11 (s, 1H), 6.86 (s, 1H), 6.91 (s, 1H), 7.33 (s, 1H), 7.77 (d, J = 8.26, 2H), 7.89 (d, J = 8.26, 2H), 8.02 (m, 2H), 8.18 (d, J = 7.75.2H), 8.90 (d, J = 8.70.2H).

The methyl compound N- [3 - ({((2S) -2- [5- (4- {4- [2 - ((2S) -1- {3 - [(methoxycarbonyl) amino] -tetrahydrofuran-3-) ilcarbonyl} tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrol-1-yl} carbonyl ) -tetrahydrofuran-3-yl] carbamate 1 (2) by the reaction of compound 1 (1) .5 and 3 - [(methoxycarbonyl) amino] tetrahydrofuran-3-carboxylic acid, LCMS (M + H) ⁺ 740, ¹ H NMR ( DMSO-d ₆ , 400 MHz) δ 1.85 (m, 8H), 2.75 (m, 2H), 3.10 (m, 2H), 3.55 (m, 2H), 3.65 (s, 6H) 3.85 (m, 2H), 4.0 (m, 4H), 4.15 (d, J = 8.75, 2H), 4.45 (d, J = 8.60.2H), 4.95 (m, 2H), 6.86 (s, 1H), 7.35 (s, 1H), 7.75 (m, 2H), 7.90 (m, 2H), 8.63 (m, 4H).

In a similar manner, methyl N - [(3S) -3 - ({((2S) -2- [5- (4- {4- [2 - ((2S) -1 - {(38) -3 - [- methoxycarbonyl) ) amino] -tetrahydrofuran-3-ylcarbonyl} tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H -pyrrol-1-yl} carbonyl) -tetrahydrofuran-3-yl] carbamate 1 (3), LCMS (M + H) ⁺ 740.

Example 2. General method for producing naphthalene-1,5-disulfonates of stereoisomers 1.1.1-1.1.10. 5 g of the corresponding base of the formulas 1.1.1-1.1.10 are dissolved in 150 ml of methanol. The resulting solution was filtered through Celite and 1.05 equiv. a solution of naphthalene-1,5-disulfonic acid tetrahydrate in methanol at the rate of 50 ml per 5 g of base. The mixture is stirred for 45-60 minutes (but no more, since with increasing mixing time the precipitate becomes extremely finely divided), the precipitate is separated by centrifugation, washed twice with methanol, ether (after each washing, the precipitate is centrifuged), transferred with ether to a round bottom flask and dried on rotary evaporator and then in a freeze dryer. Naphthalene-1,5-disulfonates of stereoisomers 1.1.1-1.1.10 are obtained with a yield of higher than 75% and a purity of 98.2% to 98.8% according to the HPLC method.

If the purity of the target product is less than 98.5%, an additional purification is carried out by repeated planting of naphthalene-1,5-disulfonate. For this, salt 1.1.1-1.1.10 is again transferred to the base. To do this, it is suspended in 100 ml of ethanol, 100 ml of a 10% sodium carbonate solution are added. After complete dissolution, add 400 ml of dichloromethane. The organic layer is separated, washed once with water and a saturated solution of sodium chloride, dried over sodium sulfate, evaporated in vacuo at a temperature not exceeding 40 ° C. Next, naphthalene-1,5-disulfonate 1.1.1-1.1.10 is planted in the above manner.

To obtain naphthalene disulfonate with a purity of more than 99%, the base is purified by column chromatography. Eluent dioxane: toluene = 1: 1.5. Combine the middle fractions, evaporate them in vacuum at a temperature not exceeding 40 ° C. Next, naphthalene-1,5-disulfonate is planted as described above. Yield 60%.

The obtained naphthalene-1,5-disulfonates are white crystalline substances with a yellow tint. Their purity and structure was confirmed by LC MS and ¹ H NMR. According to LC MS, the content of the main substance in analytical samples is> 99%. Salts and bases in the LC MS spectra have the same molecular ions corresponding to the molecular weight of the bases: 1.1.1: LC MS m / e 711 (M + 1) ⁺ . 1.1.2: LC MS m / e 711 (M + 1) ⁺ . 1.1.3: LC MS m / e 711 (M + 1) ⁺ . 1.1.4: LC MS m / e 711 (M + 1) ⁺ . 1.1.5: LC MS m / e 711 (M + 1) ⁺ . 1.1.6: LC MS m / e 767 (M + 1) ⁺ . 1.1.7: LC MS m / e 683 (M + 1) ⁺ . 1.1.8: LC MS m / e 743 (M + 1) ⁺ . 1.1.9: LC MS m / e 739 (M + 1) ⁺ . 1.1.10: LC MS m / e 739 (M + 1) ⁺ .

1.1.1: ¹ H NMR (DMSO-d ₆ , 400 MHz) 5 1.00 (m, 12H), 1.70 (m, 4H), 1.85 (m, 4H), 2.15 (m, 2H), 3.30 (t, J ¹ = 10.60, J ² = 7.30, 2H), 3.56 (t, J ¹ = 6.70, J ² = 6.92, 2H), 3.61 (s, 6H), 4.0 (m, 2H), 4.70 (m, 2H), 6.86 (s, 1H), 7.33 (s, 1H), 7.77 (m, 2H), 7.89 (m, 6H), 7.95 (m, 2H), 8.02 (m, 2H), 8.18 (m, 2H), 8.90 (m, 2H);

1.1.2: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.00 (m, 12H), 1.70 (m, 4H), 1.85 (m, 4H), 2.15 (m, 2H), 3.30 (t, J ¹ = 10.60, J ² = 7.30.2 H), 3.56 (t, J ¹ = 6.70, J ² = 6.92, 2 H), 3.61 (s, 6 H), 4.0 (m, 2 H), 4.70 (m, 2 H), 6.86 (s, 1H), 7.33 (s, 1H), 7.77 (m, 2H), 7.89 (m, 6H), 7.95 (m, 2H), 8.02 (m, 2H), 8.18 (m, 2H), 8.90 (m, 2H);

1.1.3: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.00 (m, 12H), 1.70 (m, 4H), 1.85 (m, 4H), 2.15 (m, 2H), 3.30 (t, J ¹ = 40.60, J ² = 7.30.2 H), 3.56 (t, J ¹ = 6.70, J ² = 6.92, 2 H), 3.61 (s, 6 H), 4.0 (m, 2 H), 4.70 (m, 2 H), 6.86 (s, 1H), 7.33 (s, 1H), 7.68 (m, 6H), 7.77 (m, 2H), 7.95 (m, 2H), 8.02 (m, 2H), 8.18 (m, 2H), 8.90 (m, 2H);

1.1.4: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.00 (m, 12H), 1.70 (m, 4H), 1.85 (m, 4H), 2.15 (m, 2H), 3.30 (t, J ¹ = 10.60, J ² = 7.30, 2H), 3.56 (t, J ¹ = 6.70, J ² = 6.92, 2H), 3.61 (s, 6H), 4.0 (m, 2H), 4.70 (m, 2H), 6.86 (s, 1H), 7.33 (s, 1H), 7.68 (m, 6H), 7.77 (m, 2H), 7.95 (m, 2H), 8.02 (m, 2H), 8.18 (m, 2H), 8.90 (m, 2H);

1.1.5: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.00 (m, 12H), 1.70 (m, 4H), 1.85 (m, 4H), 2.15 (m, 2H), 3.30 (t, J ¹ = 40.60, J ² = 7.30.2 H), 3.56 (t, J ¹ = 6.70, J ² = 6.92, 2 H), 3.61 (s, 6 H), 4.0 (m, 2 H), 4.70 (m, 2 H), 6.86 (s, 1H), 7.33 (s, 1H), 7.77 (m, 2H), 7.89 (m, 6H), 7.95 (m, 2H), 8.02 (m, 2H), 8.18 (m, 2H), 8.90 (m, 2H);

1.1.6: ¹ NMR (DMSO-d ₆ , 400 MHz) δ 0.88 (br.s, 3H), 0.99 (br.s, 9H), 1.32 (br.s, 12H), 1.65 (m, 2H), 1.85 (m, 2H), 1.99 (m, 3H), 2.10 (m, 3H), 3.30 (t, J ¹ = 10.60, J ² = 7.30.2H), 3.56 (m, 1H), 3.63 (m, 1H ), 3.99 (m, 2H), 4.68 (m, 4H), 6.86 (s, 1H), 7.33 (s, 1H), 7.76 (d, J = 8.26, 2H), 7.90 (d, J = 8.26.2H ), 8.02 (m, 2H), 8.18 (d, J = 7.75.2H), 8.90 (d, J = 8.70.2H);

1.1.7: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.52 (s, 6H), 1.65 (s, 6H), 1.65 (m, 2H), 1.85 (m, 2H), 2.05 (m, 4H ), 3.61 (s, 6H), 3.70 (m, 2H), 3.89 (m, 2H), 5.00 (m, 2H), 5.63 (s, 1H), 5.94 (s, 1H), 6.86 (s, 1H) 7.33 (s, 1H), 7.77 (d, J = 8.26, 2H), 7.89 (d, J = 8.26.2H), 8.02 (m, 2H), 8.18 (d, J = 7.75.2H), 8.90 ( d, J = 8.70.2 H);

1.1.8: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.56 (s, 3H), 1.65 (s, 3H), 1.65 (m, 2H), 1.85 (m, 2H), 2.05 (m, 4H ), 3.43 (s, 3H), 3.51 (s, 3H), 3.61 (s, 6H), 3.66 (m, 2H), 3.75 (br.s, 2H), 3.89 (m, 2H), 4.11 (br. s, 2H), 5.00 (m, 2H), 6.11 (s, 1H), 6.86 (s, 1H), 6.91 (s, 1H), 7.33 (s, 1H), 7.77 (d, J = 8.26, 2H) 7.89 (d, J = 8.26.2H), 8.02 (m, 2H), 8.18 (d, J = 7.75.2H), 8.90 (d, J = 8.70.2H);

1.1.9: ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 1.64 (m, 2H), 1.84 (m, 2H), 1.99 (m, 2H), 2.10 (m, 2H), 2.73 (m, 2H ), 3.10 (m, 2H), 3.56 (t, J ¹ = 6.70, J ² = 6.92.2 H), 3.61 (s, 6H), 3.90 (m, 4H), 3.98 (m, 2H), 4.14 (br s, 2H), 4.42 (broad s, 2H), 5.00 (m, 2H), 6.86 (s, 1H), 7.33 (s, 1H), 7.76 (d, J = 8.26.2H), 7.89 (d , J = 8.26.2 H), 8.02 (m, 2H), 8.18 (d, J = 7.75, 2H), 8.90 (d, J = 8.70, 2H).

Example 3. Determination of the antiviral activity of the compounds of General formula 1 and 1.1 and formulas 1.1.1-1.1.10 (inhibitory ability of the protein NS5A)

The inhibitory ability of the compounds of formulas 1, 1.1 and formulas 1.1.1-1.1.10 of the NS5A protein was determined in the Huh7 human hepatoma cell line containing the subgenomic HCV RNA replicon (genotype 1b, Conl clone). As an experimental technique, a variant of the IFA immunoassay for the viral protein NS5A was used in a 96-well format. Cytotoxicity of the compounds was evaluated in parallel.

Huh7 cells were seeded in 96-well plates (7.5 × 10 ³ cells per well in 100 μl of culture medium), solutions of the test compounds in DMEM ((DMEM) 1X; Source: Cellgro; Catalog: 10-013-CV) were prepared immediately before using. In total, eleven serial triplicate dilutions were prepared with a concentration of 20 nM to 0.2 pM. 4 hours after plating of cells, serial dilutions of the preparations were added to the cells (100 μl per well). The final concentration of the tested compounds ranged from 10 nM to 0.1 pM, and DMSO - 0.5%. If necessary, higher concentrations of the tested bis-azoles were investigated. Each dilution of the drug was tested on two identical wells. The cells were then incubated for three days at 37 ° C / 5% CO ₂ . Cells were fixed by adding 250 μl / well of acetone / methanol (1: 1). After 1 minute, the cells were washed three times with PBS (Phosphate Buffered Saline). After that, the cells were blocked by adding 150 μl / well of 10% fetal calf serum in PBS for 1 hour at room temperature. The cells were then incubated with murine monoclonal antibodies to HCV core antigen, clone C7-50 (Source: Affinity BioReagents; Catalog: MA1-080) (100 μl / well, working dilution 1: 500 in 10% fetal calf serum in PBS solution ) for two hours at 37 ° C. Cells were washed 6 times with PBS / 0.05% Tween 20 solution, after which they were incubated for 1 hour with goat antibodies to mouse immunoglobulins (conjugated with horseradish peroxidase, 100 μl / well, working dilution 1: 2500 in 10% fetal calf serum in solution PBS). Cells were washed 6 times with PBS / 0.05% Tween 20 solution, once with PBS solution, after which 100 μl / well of substrate was added (1 tablet of OPD + 12 ml of citrate / phosphate buffer + 5 μl of 30% H ₂ O ₂ ). The plates were held for 30 minutes in the dark at room temperature. The reaction was stopped by adding 100 μl / well 2 NH ₂ SO ₂ and the optical density (wavelength 490 nm) was measured using a Victor3 V 1420 multichannel spectrophotometer (Perkin Elmer). IC ₅₀ values (azole concentration that lowers the level of viral RNA replicon by 50%) for each bis-azole tested were calculated using the XLfit 4 program.

The cytotoxicity of compounds of the general formula 1, 1.1 and formulas 1.1.1-1.1.10 was studied in experiments on the culture of the Huh7 human hepatoma cell line. The number of living cells was determined using the ATPLite kit (Perkin Elmer, Boston, USA) in accordance with the manufacturer's instructions. The cytotoxic effect was evaluated by plating cells in a black microplate with a transparent bottom (96 cells, 104 cells per well). Three independent repeats were used for each bis-azole. Test bis-azoles were added after 18 hours, after which the cells were incubated with substances for 96 hours. Washed twice each well with phosphate-buffered saline (0.2 ml / moon) and then lysed the cells by adding cell buffer (50 μl / moon) (all of these reagents are included in the ATPLite kit). The microplate was incubated for 5 minutes on a rotating platform at 600 rpm, after which 50 μl of substrate solution (part of the ATPLite kit) was added to each well. Incubated for another 5 minutes on a rotating platform at 600 rpm, kept for 10 minutes in the dark and then measured the luminescence on a TopCount NXT instrument (Packard, Perkin Elmer). As a quantitative parameter for assessing cytotoxicity, we used the value of CK ₅₀ , which corresponds to the concentration of bis-azole, at which 50% of the cells die. Calculation of the CK ₅₀ parameter: to calculate the inhibition efficiency (% Ing), we used the formula:% Ing = [(L ^pos -L ^ex ) / L ^pos -L ^neg )] * 100%, where L ^pos is the positive control, luminescence in cells with cells without substance; L ^neg - negative control, luminescence in cells with a medium without cells; L ^ex - luminescence in cells with a substance in a certain concentration. The values of CC _{50 were} then calculated using the XLfit 4 program.

The test results of the new compounds of the general formula 1, 1.1 and formulas 1.1.1-1.1.10 indicate their high picomolar activity. The inhibitory activity in relation to HCV genotype 1b, 1a and 2a of the new compounds is presented in table 1 and is designated as *> 1000 nM, ** from 999 nM to 10 nM, *** from 9.9 nM to 1 nM and **** < 1 nM

Table 1 Inhibitory activity towards HCV genotype 1b of compounds of general formula 1.1.1 and formulas 1.1.1-1.1.10. Compound gT1b gT1a gT2a FBS EC ₅₀ , nM 1 (1) ** * 1 (2) *** * * 1 (3) *** ** 1.1.1 ** * * 1.1.2 ** * 1.1.3 **** ** *** 1.1.4 **** *** *** 1.1.5 *** *** ** 1.1.6 ** * * 1.1.7 ** * 1.1.8 ** * * 1.1.9 *** * * 1.1.10 *** * **

Example 4. A study of the storage of naphthalene 1,5-disulfonates

All samples were stored in glass vials, sealed with rubber stoppers with aluminum caps, in a refrigerator at a temperature of ~ 4.7 ° C and in a freezer at a temperature of about -23 ° C.

The amount of impurities was determined by the internal normalization method; detection was carried out by a UV detector at a wavelength of 220 nm.

Below are the test results of naphthalene-1,5-disulfonate of compound 1.1.4, indicating its satisfactory stability at low temperature.

Example 5. Obtaining a pharmaceutical composition in the form of tablets

1600 mg of starch, 1600 mg of ground lactose, 400 mg of talc and 1000 mg of compound 1.1.4 are mixed. The resulting bar is crushed into granules and sieved through sieves, collecting granules with a size of 14-16 mesh. The granules obtained are tabletted into a suitable tablet form weighing 560 mg each.

Example 6. Obtaining a pharmaceutical composition in the form of capsules

Compound 1 (1) is thoroughly mixed with lactose powder in a 2: 1 ratio. The resulting powder mixture is packaged in 300 mg in a suitable size gelatin capsule.

Example 7 Preparation of a Pharmaceutical Composition in the Form of Injectable Compositions for Intramuscular, Intraperitoneal, or Subcutaneous Injection

500 mg of compound 1.1.4 are mixed with 300 mg of chlorobutanol, 2 ml of propylene glycol and 100 ml of injection water. The resulting solution is filtered and placed in 1 ml ampoules, which are sealed.

Claims (7)

1. Alkyl [2- (2- {5- [4- (4- {2- [1- (2-methoxycarbonylamino-acetyl) pyrrolidin-2-yl] -3H-imidazol-4-yl} phenyl) -buta-1,3-diinyl] -1H-imidazol-2-yl} -pyrrolidin-1-yl) -2-oxoethyl] carbamate of the general formula 1

where R ¹ , R ² , R ³ and R ⁴ independently from each other represent C ₁ -C ₃ alkyl;
R ⁵ and R ^6, independently of one another, are C ₁ -C ₃ alkyloxymethyl, or
R ³ _, and R ⁵ _, and R ⁴ _, and R ^6, together with the carbon atoms to which they are bonded, independently form a tetrahydrofuran ring. 2. Alkyl [2- (2- {5- [4- (4- {2- [1- (2-methoxycarbonylamino-acetyl) -pyrrolidin-2-yl] -3H-imidazol-4-yl} -phenyl) -buta-1,3-diinyl] -1H-imidazol-2-yl} -pyrrolidin-1-yl) -2-oxoethyl] carbamate naphthalene-1,5-disulfonates of the general formula 1.1

Where
R ¹ , R ² , R ³ and R ^{4 are} independently C ₁ -C ₃ alkyl;
R ⁵ and R ^{6 are} independently C ₁ -C ₃ alkyl or C ₁ -C ₃ alkyloxymethyl, or
R ³ _. and R ⁵ _, and R ⁴ _, and R ^6, together with the carbon atoms to which they are attached, independently form a tetrahydrofuran ring, excluding alkyl [(S) -1 - ((S) -2- {5- [4 - (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1, 3-diinyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate. 3. Compounds representing:
methyl [(R) -1 - ((R) -2- {5- [4- (4- {2 - [(R) -1 - ((R) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate ( 1.1.1 );
methyl [(S) -1 - ((R) -2- {5- [4- (4- {2 - [(R) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate ( 1.1.2 );
methyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate ( 1.1.3 );
methyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((S) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate ( 1.1.4 );
methyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((R) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate ( 1.1.5 );
isopropyl [(R) -1 - ((S) -2- {5- [4- (4- {2 - [(S) -1 - ((R) -2-methoxycarbonylamino-3-methyl-butyryl) - pyrrolidin-2-yl] -3H-imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-carbonyl) -2-methyl-propyl] carbamate naphthalene-1,5-disulfonate ( 1.1.6 );
methyl [2 - ((S) -2- {5- [4- (4- {2 - [(S) -1- (2-methoxycarbonylamine-2-methyl-propionyl) pyrrolidin-2-yl] -3H -imidazol-4-yl} -buta-1,3-diynyl) phenyl] -1H-imidazol-2-yl} pyrrolidin-1-yl) -1,1-dimethyl-2-oxoethyl] carbamate naphthalene-1,5-disulfonate ( 1.1.7 );
methyl N- [1 - ({(2S) -2- [5- (4- {4- [2 - ((2S) -1- {2 - [(methoxycarbonyl) amino] -2-methyl-3-methoxypropioyl } tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrol-1-yl} carbonyl) -1-methyl-2-methoxyethyl] carbamate naphthalene-1,5-disulfonate ( 1.1.8 );
methyl N- [3 - ({(2S) -2- [5- (4- {4- [2 - ((2S) -1- {3 - [(methoxycarbonyl) amino] -tetrahydrofuran-3-ylcarbonyl} tetrahydro -1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrol-1-yl} carbonyl) -tetrahydrofuran -3-yl] carbamate naphthalene-1,5-disulfonate ( 1.1.9 );
methyl N - [(3S) -3 - ({(2S) -2- [5- (4- {4- [2 - ((2S) -1 - {(3S) -3 - [(methoxycarbonyl) amino] -tetrahydrofuran-3-ylcarbonyl} tetrahydro-1H-pyrrol-2-yl) -1H-imidazol-5-yl] -1,3-butadiinyl} phenyl) -1H-imidazol-2-yl] tetrahydro-1H-pyrrole- 1-yl} carbonyl) -tetrahydrofuran-3-yl] carbamate naphthalene-1,5-disulfonate ( 1.1.10 ). 4. NS5A protein inhibitors. representing compounds of general formula 1, or of general formula 1.1. or formulas according to any one of claims 1 to 3. 5. A pharmaceutical composition for treating and preventing diseases caused by hepatitis C virus and GBV-C hepatitis virus, comprising, as an active component, an NS5A protein inhibitor according to claim 4 in a therapeutically effective amount. 6. A drug in the form of tablets, capsules or injections in a pharmaceutically acceptable package for the treatment and prevention of diseases caused by hepatitis C virus and GBV-C hepatitis virus, containing, as an active component, an NS5A protein inhibitor according to claim 4 or a pharmaceutical composition according to claim 5 in a therapeutically effective amount. 7. A method for the prevention and treatment of diseases caused by hepatitis C virus and hepatitis GBV-C virus by administering a therapeutically effective amount of a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 5, or a drug according to claim 6.