BR102012020638A2 - MIMETIC PEPTIDE COMPOUNDS DERIVED FROM TARTARIC ACID POTENTIALLY ACTIVE AGAINST HEPATITIS C VIRUS AND PHARMACEUTICAL COMPOSITION CONTAINING SUCH COMPOUNDS - Google Patents

Abstract

MIMETIC PEPTIDE COMPOUNDS DERIVED FROM POTENTIALLY ACTIVE TARTARIC ACID AGAINST HEPATITIS CE VIRUS PHARMACEUTICAL COMPOSITION CONTAINING SUCH COMPOUNDS The present invention relates to tartaric acid-derived mimetic compounds potentially active against hepatitis C viruses and pharmaceutical compositions containing such compounds as synthetic Hepatitis C virus (HCV) polymerase and serine protease, characterized by having a structure of the 1,4: 3,6-dianhydromanitol type and a tartaric acid type meat. The side portions are characterized by mimetic peptide bonds from coupling with various amino esters.

Description

Patent Invention Descriptive Report

Potentially Active Tartaric Acid Derivative Mimetic Peptide Compounds Against Hepatitis C Virus and Pharmaceutical Composition Containing Such Compounds

Field of the Invention

The present invention relates to POTENTIALLY ACTIVE TARTARIC ACID DIMENSIVE PEPTIDE COMPOUNDS AGAINST 10 HEPATITIS VIRUS PHARMACEUTICAL COMPOSITION CONTAINING SUCH COMPOUNDS, designed as inhibitors of HCV (HVatitis C, synthase C, synthase, Hepatitis C virus) in the central portion, a structure derived from tartaric acid. The side portions are characterized by having a 1,4: 3,6-15 dianhydromanitol type structure and mimetic peptide bonds from coupling with various amino esters. These new compounds, if active against the hepatitis C virus, could be the basis for the preparation of antiviral formulations capable of stopping HCV proliferation responsible for flavivirosis, hepatitis C.

20

Background of the Invention

Flaviviridae is a large family of viral pathogens that cause various diseases and high death rates among animals, especially humans. This family consists of three genera: Flavivirus, Pestivirus and Hepacivirus. The Flaviviridae family includes hepatitis C viruses (genus Hepacivirus), yellow fever, West Nile and dengue, which belong to the genus Flavivirus (Linderbach BD; Thiel HJ; Rice CM Flaviviridae: Viruses and their replication in Fields Virology 2007, 33, 1101-1133).

Hepatitis is an inflammatory disease that compromises hepatocytes and can be classified as viral, toxic (caused by reaction to alcohol, drugs or drugs), or autoimmune, a condition in which cytotoxic cells act on liver tissue (Manual Advice on Viral Hepatitis / Health Surveillance Secretariat, Department of Epidemiological Surveillance, Brasília, DF, 2005).

In the case of viral hepatitis, seven types have already been characterized: A, B, C, D, E, G and TT, which have in common hepatotropism and may lead to inflammation and liver necrosis. Viral hepatitis, in particular, is caused by different globally distributed etiological agents, including HCV (Craxi, A.; Laffi, G .; Zignego, AL, Hepatitis C virus (HCV) infection: a systemic disease, Mol. Aspects Med. 2008, 29, 85-95; Manns, MP; Foster,

G.R .; Rockstroh, J.K .; Zeuzem, S .; Zoulim, F .; Houghton, M. Nat. Rev. Drug Discov. 2007, 6, 991-1000).

HCV causes persistent liver infections, that is, mild, non-progressive liver infections, thus without symptoms. However, in the vast majority of patients who develop the severe form of hepatitis C, the infection becomes chronic, which can result in cirrhosis, failure or liver carcinoma 15 causing thousands of deaths annually (Chevaliez S. & Pawlotsky JM HCV Genome and Iife Cycle in Hepatitis C virus genomes and molecular biology 2006, 1, 5-47).

In structural terms, HCV is an enveloped virus and has an RNA genome of approximately 9.4Kb. The genome has a single open reading window encoding four structural proteins (C, E1, E2, and p7) and six nonstructural proteins (NS2 to NS5B). Two untranslated regions (RTU) present at each end of the genome are important for the RNA genome translation and transcription steps. Based on its evolutionary history, inferred from phylogenetic analysis, HCV is classified into 6 genotypes (from 1 to 6), whose RNAs differ by more than 30% in nucleotide sequence. However, they all share an identical complement of co-linear genes of the same size in the open reading window, the main differences being found throughout the genome, especially in regions encoding the E2 envelope protein, 30 probably originating from immunologically selected mutations. (Simmonds P .; Bukh J .; Combet C .; Deléaqe G .: Enomoto N .: Feinstone S .: Halfon P .: Inchauspe G .: Kuiken C .: Maertens G .; Mizokami M .: Murphv DG; Okamoto H Pawlotskv JM: Penin F.; Sablon E .: Shin-I T .; Stuvver LJ: Thiel.

H. J .; Viazov S .: Weiner A. J .: Widell A, Consensus proposals for a unified system of nomenclature of hepatitis C virus genotypes ,. Hepatology 2005, 42 (4), 962-973). There is a different geographical distribution in relation to different genotypes. In Brazil, the most frequent are genotypes 1, 2 and 3 with an incidence of 70; 2.5 and 28%, respectively.

The viral genome encodes a single long polyprotein of

approximately 3000 amino acids. This polyprotein is further cleaved into four structural proteins (C, E1, E2 and p7) and six nonstructural (NS) proteins designated NS2, NS3, NS4A, NS4B, NS5A and NS5B. While structural proteins are processed by proteases of the host cell in the endoplasmic reticulum (ER), the nonstructural proteins are cleaved by the NS2 proteins and the NS3 / NS4A complex, which has an N-terminal proteinase domain (Grakoui A. ; McCourt DW; Wychowski C.; Feinstone SM; Rice, CM, Characterization of the hepatitis C virus-encoded serine proteinase: determination of the proteinase-dependent polyprotein cleavage 15 sites, J. Virot. 1993, 67, 2832-2843; C. Francesco RJ, Both NS3 and NS4A are required for proteolytic processing of hepatitis C virus (nonstructural proteins, Virol. 1994, 68, 3753-3760).

The most commonly used therapy for individuals with chronic hepatitis C is the combination of interferon alfa (IFN-α) injections and oral ribavirin doses. 20 Studies with patients treated with IFN-a alone showed that only 10-19% of them had sustained response, defined as the absence of detectable HCV RNA 24 weeks after therapy. When ribavirin was used in combination with IFN-a, 38-43% of treated patients (infected with genotypes 2 to 6) showed sustained virological response, which was accompanied by better histological (biopsy) results. Ribavirin is a synthetic guanosine analogue that has direct action against RNA and DNA viruses by probable mechanism of inhibition of virus-dependent RNA polymerase. It alone, however, has no effect on hepatitis C.

Combining polyethylene glycol (PEG) with IFN-α resulted

IFN-α (PEG-IFN) slower absorption and elimination. PEG-IFN is only given subcutaneously once a week and during this time the level of the drug in the blood remains constant. Another advantage of PEG-IFN monotherapy is that the sustained response, which was 10-19% with IFN-α, was 39% for patients infected with genotypes 2 to 6. However, this percentage increases with PEG-IFN + ribavirin combination, and is now considered the most effective treatment against 5 hepatitis C today (Fried MW: Shiffman ML: Reddv KR: Smith C .: Marinos G .; Goncales FL Jr .: Háussinqer P .: Diaqo M .: Carosi G .: Dhumeaux D .; Craxi A.: Lin A.: Hoffman J .; Yu J., Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection, N. Engi J. Med. 2002, 347, 975-982).

Even with this therapeutic arsenal, HCV infection accounts for over 80% of liver transplants worldwide, and in Brazil (Consensus of the Paulista Infectious Society (SPI) for Hepatitis C Management and Therapy, São Paulo, SP .). In addition, this treatment causes many side effects to patients, as both interferon and ribavirin induce a systemic antiviral response. Interferon causes fever, chills, and flu-like symptoms, while ribavirin can cause hemolytic anemia, severe depression leading to suicidal tendencies, and even birth defects. As a consequence of these effects, a study with HCV patients undergoing PEG-IFN treatment showed that approximately 35% of patients withdrew therapy (Belon, CA & Frick, DN, Helicase inhibitors as specifically targeted antiviral therapy for hepatitis C, Future Virol). 2009 4 (3), 277-293).

Thus, the need to identify drugs that target specific proteins or HCV RNA is becoming increasingly evident in order to optimize the standard treatment, allowing it to be more effective and cause fewer side effects to patients.

HCV has two potential targets for the development of new antiviral drugs. One is the NS5B RNA-dependent RNA polymerase protein (RdRp) that can replicate one RNA molecule into another. 30 This is a unique activity of RNA viruses, and this activity is not found in host cells. Thus NS5B RdRp is a very specific target for HCV. In addition to viral replicase we also have NS3-4A (serinoprotease) which is responsible for processing viral polyprotein. Despite being a serine protease, NS3 has a very different molecular structure than human cell serine proteases, which

The three-dimensional structure elucidated by X-ray crystallography and diffraction has shown that it has two β-leaf subdomains forming typical barrel 5 of a serine protease chymotrypsin. It has a substrate interaction slit located between the subdomains, important for proteolytic activity (Kim JL; Morgenstern KA; Lin C.; Fox T .; Dwyer MD; Landro JA; Chambers SP; Markland W.; Lepre CA; 0 'Malley ET; Harbeson SL; Rice CM; Murcko MA; Caron PR; Thomson JA, Crystal Structure of the Hepatitis C Virus NS3 Protease Domain Complexed with a Synthetic NS4A Cofactor Peptide, Ce //, 1996, 87, 343-355) . The NS3 protease domain is primarily responsible for self-cleavage of the HCV polyprotein NS3 / 4A site, while subsequent cleavages at the other sites occur preferably in the order NS5A / 5B, NS4A / 4B, NS4B / 5A (Steinkuhler C .; 15 Urbani A.; Tomei L .; Biasiol G .; Sardana M .; Bianchi E .; Pessi A .; From Francesco R., Activity of Purified Hepatitis C Virus Protease NS3 on Peptide Substrates, J. virol. ), 6694-6700; Steinkuhler C. Hepacivirin in Handbook of proteolytic enzymes. 2004, 1773-1779).

By comparing the NS3 sequence of other flaviviruses we can identify three conserved residues (His-57, Asp-81, and Ser-139) that make up the catalytic center of this protease. The NS3 protease active center contains a tetrahedral oxyon stabilizing site, reaction intermediate, oxyanion / stabilization loop, and an E2 tape preceded by an E1 tape.

The NS3 assumes a structure of two pleated beta tape sets. The first located in the N-terminal portion contains tapes A1, B1, C1, D1, E1, and F1. The second beta barrel consists of tapes A2, B2, C2, D2, E2, and F2.

Several papers have been reported on serine protease inhibitors. Recently, hexapeptide derivatives containing an electrophilic α-keto amide function have been described as potent inhibitors of HCV serine protease (Arasappan, A.; Njoroge, FG; Chan, T.-Y .; Bennett, F.; Bogen, SL; Chen, K .; Gu, H.; Hong, L.; Jao, E.; Liu, Y.-T.; Lovey, RG; Parekh, T.; Pike, RE; Pinto, P.; Santhanam, B. ; Venkatraman, S.; Vaccaro, H.; Wang, H .; Yang, X .; Zhu, Z .; Mckittrick, B.; Saksena, AK; Girijavallabhan, V .; Pichardo, J .; Butkiewicz1 N.; Malcolm B. Prongay A. Yao Y Marten B. Madison V Kemp S Levy O Lim-Wilby M Tamura S S; Ganguly1 AK, Hepatitis C virus NS3-4A serine protease inhibitors: SAR of P2 moiety with improved potency, Bioorg. Med. Chem. Lett. 2005, 15, 4180-4184). Still within the α-keto amide class, telaprevir and boceprevir are in clinical development. (Chen, KX; Vibulbhan, B .; Yang, W .; Cheng, KC .; Liu, R .; Pichardo1 J .; Butkiewiez1 N.; Njoroge, FG. Potent and selective small molecule NS3 serine protease inhibitors of Hepatitis C virus with dichlorocyclopropylproline as P2 residue, Bioorg. Med. Chem. 2008, 10 doi: 10.1016 / jbmc.2007.11.002. Perni, RB; Almquist, SJ; Bym, RA; Chandorkar, G .; Chaturvedi1 PR; Courtney, LF Decker, CJ; Dinehart, K.; Gates 1 CA; Harbeson, SL; Heiser, A.; Kalkeri, G .; Kolaczkowski, E.; Lin1 K .; Rao1 BG; Taylor1 WP; Thomson, JA; Tung1 RD; Wei1 Y .; Kwong1 AD; Lin1 C., Preclinical profile of VX-950, a potent, selective, andally bioavailable inhibitor of hepatitis C virus NS3-4A serine protease, Antimicrob. Agents Chemother. 2006, 50, 899-909).

According to this scenario, the major inhibitors that have been promising in human clinical studies to date focus on inhibiting proteolytic processing (viral NS3-4A protease inhibitors) and RNA genome replication (NS5B inhibitors). RNA polymerase viral dependent RNA), as shown in Table 1 below.

Table 1 - Protease inhibitors and viral polymerase in development for the treatment of hepatitis C (Asselah, T. & Marcellin1 P .. New direct-acting antivirals' combination for the treatment of chronic hepatitis C1 Liver International 2011, 31 (1), 68-77).

Drug name Manufacturer Study phase Molecular target Telaprevir Vertex - Tibotec Phase 3 NS3-4A protease Boceprevir Schering-Plow / Merck Phase 3 NS3-4A protease TMC 435350 Tibotec / Medavir Phase 2 NS3-4A protease BI 201335 Boehringer Ingelhelm Phase 2 NS3- 4A protease R 7227 / ITMN 191 InterMune / Roche Phase 2 NS3-4A protease MK 7009 Merck Phase 2 NS3-4A protease BMS 650032 Bristol-Myers Squibb Phase 1 NS3-4A protease PHX 1766 Phenomix Phase 1 NS3-4A protease R 7128 Roche / Pharmasset Phase 2 NS5B RdRp PS -7851 Pharmasset Phase 1 NS5B RdRp IDX 184 Idenix Phase 1 NS5B RdRp BI 207127 Boehringer Ingelhelm Phase 2 NS5B RdRp GS 9190 Gilead Phase 1 NS5B RdRp GS 9190 Gilead Phase 1 NS5B RdRp ViroChem Pharma Phase 1 NS5B RdRp VCH 916 Vi roChem Pharma Phase 1 NS5B RdRp VCH 222 ViroChem Pharma Phase 1 NS5B RdRp ANA 598 Anadys Phase 1 NS5B RdRp ABT 333 Abbott Phase 1 NS5B RdRp The targets of anti-HCV therapy are concentrated in the different stages of viral RNA replication, and in addition to these. New therapies are targeted at the translation steps (ribozymes, antisense oligos, RNAi), proteolytic processing (viral protease inhibitors) and RNA genome replication (viral RNA polymerase inhibitors).

In recent studies, the telaprevir viral protease inhibitor has been used as a third drug in the treatment of patients with chronic hepatitis C (infected with genotype 1), along with PEG interferon and Ribavirin, in order to obtain a greater therapeutic response. These studies demonstrated that treatment with the addition of telaprevir increased the rapid virologic response from 11% to 79% and the sustained virologic response from 35% to 61% compared to standard treatment patients (McHutchison JG; Everson GT; Gordon SC; Jacobson IM; Sulkowski M .; Kauffman R.; McNair L .; Alam J .; Telaprevir with peginterferon and ribavirin for chronic HCV genotype 1 infection, N Engl J Med. 2009, 360, 1827-1838; Hézode C .; Forestier N .; Dusheiko G .: Ferenci P .; Pol S .; Goeser T. Bronowicki JP; Bourlière M .; Gharakhanian S .; Benqtsson L .; Georqe S .; Kieffer T. ; Kwonq A .; Kauffman RS; Alam J .; Pawlotskv JM; Zeuzem S., Telaprevir and peginterferon with or without ribavirin for chronic HCV infection, N Engl J Med. 2009, 360, 1839-1850).

Another protease inhibitor that is also being tested is boceprevir, which when combined with standard treatment has shown a 70% increase in sustained virological response in patients with chronic hepatitis C (genotype 1 infected) (Kwo PY; Lawitz EJ ; McCone J .; Schiff ER: Vierlinq JM; Pound P .; Davis MN; Galati JS; Gordon SC; Ravendhran N .: Rossaro L.; Anderson FH; Jacobson IM;

Rubin R .; Kourv K .; Pedicone L. P .; Brass C. A .; Chaudhri E .; Albrecht JK Efficacy of boceprevir, NS3 protease inhibitor, combination with peginterferon alfa-2b and ribavirin in treatment-naive patients with genotype 1 hepatitis C infection (SPRINT-1): open-label, randomized, multicentre phase 2 trial, Lancet, 2010, 376, 705-716). Both inhibitors are in phase 3 clinical trials in

stressing the importance of NS3-4A protease as one of the main targets for the treatment of chronic hepatitis C patients. However, despite increased treatment efficacy, both telaprevir and boceprevir cause side effects to patients, such as anemia, 15 nausea, headaches, and insomnia, thus indicating the need for more in-depth testing and more patients. until these drugs are effectively added to standard HCV treatment (Asselah, T. & Marcellin, P., New direct-acting antivirals' combination for the treatment of chronic hepatitis C1 Liver International 2011, 31 (1), 68-77) .

In the patent literature, documents related to

but without infringing its scope, the most relevant being described below:

WO2008137126 describes a method of treating HCV infection using a therapeutic combination comprising a serine protease inhibitor and a polymerase inhibitor. Not being a single compound as inhibitor of both HCV serine protease and polymerase, such disclosed compounds are part of the state of the art.

WO2007092616 describes pharmaceuticals, compositions or pharmaceutical kits and methods, based on combinations of at least one HCV protease inhibitor and one HCV polymerase inhibitor, to be administered in the treatment of symptoms caused by HCV. Compounds that act as protease and polymerase inhibitors are also described.

US7105499 describes compounds that act as HCV polymerase inhibitors. US2007027071 describes macrocyclic compounds that act as HCV protease inhibitors.

UFRJ1 owned document PI0401908-3 describes synthetic serine protease inhibitor compounds having a C2 axis of symmetry, used as a basis for the preparation of antiviral formulations capable of stopping the proliferation of viruses responsible for most flaviroses, such as , hepatitis C, Dengue and West Nile fever.

As can be seen there are several documents describing either protease inhibitors or polymerase inhibitors, or a combination of both, but neither can describe a compound capable of acting as a protease and polymerase inhibitor, more specifically related to HCV.

Summary of the Invention

The first object of the present invention relates to serine protease inhibiting compounds and potentially inhibitors of RNA and DNA polymerases of formula (I):

The second object of this invention relates to a pharmaceutical composition containing said compound of formula (I),

H OR2 O O

20

(I)

H OR2 O O

25

(I) as well as their derivatives and / or pharmaceutically acceptable salts thereof, for the manufacture of a medicament used as inhibitors of DNA and RNA polymerases and / or serine proteases.

The third object of this invention is a composition

pharmaceutical compound containing the compound of molecular formula (I),

(I)

as well as derivatives and / or pharmaceutically acceptable salts thereof for the manufacture of a medicament used for the treatment of diseases caused by the Flaviviridae family viruses in human and non-human mammals.

Description of the Figures Figure 1: Compound 12a3 dose-response to activity

HCV-1b NS3 / 4A protease proteolytic. The points represent the averages of relative activities for each compound concentration (0, 12.5, 25, 50, 100, 200, and 400μΜ) from two independent experiments.

Detailed Description of the Invention

The present invention relates to compounds of formula (I), designed as polymerase and serine protease inhibitors.

These compounds are potentially active in flaviviridae viruses, preferably; Such compounds are active in inhibiting HCV serine protease and dengue virus.

For the polymerase and serine protease inhibiting compounds of the present invention of formula (I), JT R OR1

(I)

R is defined as amino esters derived from the amino acids Lfenylalanine (L-Phe), L-valine (L-VaI) 1 L-glycine (L-Gly), L-proline (L-Pro) 1 L5 arginine (L-Arg) and L-Lysine (L-Lys) which will be used in the synthesis of pseudopeptides. Z being benzyl (-Bn), carboxybenzyl (-Cbz) or carboxy-t-butyl (-Boc); capable of imparting rigidity to the inhibitor, and which by their nature may also act on polymerases, such as DNA or RNA polymerases, by inhibiting them.

Said compounds of molecular formula (I) 1 as well as their

Pharmaceutically acceptable derivatives and / or salts are used in a pharmaceutical composition as inhibitors of DNA and RNA polymerases and serine proteases.

Such pharmaceutical composition has a concentration ranging from 10μΜ to 400μΜ of said compound of molecular formula I, as well as pharmaceutically acceptable salts and / or derivatives thereof.

For purposes of this invention the excipients are all those chemical compounds which are intended to give the pharmaceutical composition form, volume, stability. All excipients to be used herein are those from the pharmaceutical industries such as adjuvants, sweeteners, solvents, stabilizers, antioxidants, preservatives, buffers, flavorants, surfactants, humectants, lubricants, dispersants and all those known to one skilled in the art.

Such excipients may be employed in different pharmaceutical forms such as powders, tablets, capsules, nanocapsules, nanospheres, hot melt extrusion processes, co-crystals, ampoules, oral solutions, transdermal patches, muscle or parenteral injectable solutions, ointments. , creams, gels, suspensions, emulsions, aerosols, dyes, elexirs, pastes, pastilles, suppositories, urges, eggs, and all those known to one of ordinary skill in the art. Such a pharmaceutical composition containing the compounds (I), as well as their pharmaceutically acceptable salts and derivatives, is employed for the manufacture of a medicament in the treatment of diseases caused by the Flaviviridae viruses, viruses of this family comprising hepatitis C viruses. dengue fever, yellow fever, west fever or Nile fever, and is preferably used to treat diseases caused by dengue and hepatitis C virus. In the case of hepatitis C the following viruses A, B, C, D, E, G and TT.

Compounds of formula (I) were produced from isomanide 10 as analogues to those previously published by the group, with the additional analogy to pro-nucleosides, that is, compounds capable of inhibiting polymerases. Its structural analogy with cyclic rigid dipeptides makes it an excellent chassis, rigid core, scaffold. Moreover, because it has a C2 axis of symmetry, it can be homologated, in two or in one position, maintaining or not the symmetry axis, via reaction with different amino acids and dihydroamino acids, for example, via direct coupling catalyzed by EDC1 HOBt, and amino acids.

From the data obtained above, the synthesis step of the predicted serine protease inhibitors was then passed.

The first step in obtaining derivatives of formula (I)

It consisted of the conversion of isomanide (1) to the mono-tosylated derivative 2 by reaction with p-toluenesulfonyl chloride in pyridine for 12h at room temperature. Treatment of mono-tosylate 2 with benzyl chloride in basic medium using TBAB as the phase transfer catalyst provided the benzylated derivative 3. This reacted with sodium azide in [bmim] BF4 at 120 ° C for 12h and provided the azide. 4, with inversion of configuration. Azide Reduction

4 with palladium on carbon in ethanol for 12h using a pressure of 40psi resulted in amine 5 (Scheme 1) (Barros, TG; Pinheiro, S.; Williamson, JS; Tanuri, A.; Pereira, HS; Toast, RM; Alonso Neto, JB; Antunes, O. 30 BC; Muri, EMF; Synthesis. 2009, 620. Barros, TG; Pinheiro, S.; Williamson, JS; Tanuri, A.; Pereira, HS; Toast, RM; Alonso Neto, JB; Antunes, OAC; Muri, EMF; Amino acids. 2010, 3, 701).

Different solvents may be used in the nucleophilic substitution step, such as DMF and DMSO. However, [bmim] BF4, ionic liquid, 10

15

It has many advantages over conventional organic solvents and has a better reaction yield. Ionic liquids have no measurable vapor pressure at room temperature, and even at very high temperatures they only degrade above 400 ° C and can be used in a vacuum without solvent loss.

The [bmim] BF4 9 ionic liquid was synthesized from N-methyl imidazole (7). This reacted with 1-bromobutane to derivative 8 which upon ion exchange with NaBF4 gave 7. The ionic liquid was purified by silica gel chromatographic column eluting with dichloromethane and was obtained in 83% yield as shown in the scheme. 2.

HO 1 you.

BnCl, TBAB 50% NaOH CH 2 Cl 2

H

OTs 75%

OrV

H

OTs

3

'NaN3

[bmimj + fBFJ "H2.5% Pd / C BnQ4.0> 73%

EtOH

. o-l

quantitative

Scheme 1. Synthesis of intermediate compounds to obtain the amine 5.

\

N

1) Bromobutane, Toluene, O0C

2) 10 O, 36h

N

7th

NaBF 4, CH 2 Cl 2 24h, r.t.

- »

83%

(CH2) 3CH3 8

Scheme 2. Synthesis of ionic liquid [Bmim] + [BF4] '. Compounds containing non-hydrolyzable tartaric acid heartwood have been described as inhibitors of HIV-1 protease (Peçanha, EP; Figueiredo, LJO; Toindeiro, RM; Tanuri, A.; Calazans, AR; Antunes, OAC; Il Farmaco. 2003, 58 149. Barros JC da Silva JFM Calazans A. Tanuri A;

Toast, R. M .; Williamson, J. S .; Antunes, O. A. C .; Lett. Org. Chem. 2006, 3,882. Resende, G. O .; Aguiar, L. C. S .; Cotrim, B.A .; da Silva, J. F. C .; Antunes, O. A. C .; Lett. Org. Chem. 2007, 4, 168). Mimetic peptides derived from L-tartaric acid have already been synthesized by the classical protocol, EDC / HOBt from free acid, or using the corresponding acid chloride, forming pseudo-peptides with a C2 symmetry axis (Peçanha, EP; Figueiredo, LJO; Toast, RM; Tanuri, A.; Calazans, AR; Antunes, OAC; Il Farmaco. 2003, 58, 149. Barros, JC; da Silva, JFM; Calazans, A.; Tanuri, A.; Toast, RM ; Williamson, JS; Antunes, OAC; Lett. Org. Chem. 2006, 3,882).

An efficient synthetic route to obtaining mimetic peptides

containing the tartaric acid heartwood but without a symmetry C2 axis is shown in scheme 3, starting from diacetyl Ο, Ο-tartaric anhydride (10a-b), which is directly prepared respectively from acids D- and L- Tartaric acid in good yields (Shrlner, RL; Furow, CL Org. Synth. Coll. 4: 242, 1963. 20 Resende, GO; Aguiar, LCS; Cotrim, BA; da Silva, JFC; Antunes, OAC; Lett. Org. Chem 2007, 4, 168). D-tartaric diacetyl β, β-anhydride (10a) reacted with L-glycine ethyl ester hydrochlorides, L-phenylalanine methyl esters and L-valine in the presence of N-methyl morpholine (NMM) to form the corresponding pseudo 11a1-a3 peptides in moderate yields. The reaction with L-proline methyl ester hydrochloride was unsuccessful so product 11a4 was obtained using the free amine, L-proline methyl ester. Similarly, diacetyl β, β-tartaric anhydride (10b) reacted with the methyl ester hydrochlorides of L-phenylalanine and L-valine by providing 11b2-b3 acids. Compound 11b4 was also obtained using the free amine of L-proline methyl ester (Scheme 3) (Resende, GO; Aguiar, LCS; Cotrim, BA; da Silva, JFC; Antunes, OAC; Lett. Org. Chem. 2007 , 4, 168).

Thus, a coupling reaction of acids 11a-b and amine 5 prepared according to scheme 1 using the classic EDC / HOBt / NMM protocol provided the corresponding 12a-b mimetic peptides in moderate yields (Scheme 3).

In order to verify the importance of acetyl groups in the biological activity of synthesized compounds, compounds 12b2-b4 were selectively unprotected under basic conditions. Selective methanolysis of the acetyl groups was obtained by treating said compounds with K2 CO3 in methanol to obtain the 13b2-b4 diols in good yields. (Resende, G. O .; Aguiar, L. C. S.; Cotrim, B. A.; da Silva, J. F. C.; Antunes, O. C. C.; Lett. Org. Chem. 2007, 4, 168).

AcO OAc

10a-b

OAeO O

HO2C R ^ OR1

OAe

Wai: R = -NHCH2, R1 = Et IIa2-b2: R = -NHCH2Bnj R1 = Me IIa3-b3: R = -NHCH2-i-Pr, R1 = Me Ua4-b4: R = -NH (CH2) 3- , R1 = Me

X R OR1

12al: R = -NHCH2, R1 = Et 12a2-b2: R = -NHCH2Bn, R1 = Me 12a3-b3: R = -NHCH2-i-Pr, R1 = Me 12a4-12b4: R = -NH (CH2) 3 -, R1 = Me

H OH O

Qxx? ' R oRi

H · '} - The OH ΒηΟ’Ό0

13b2: R = -NHCH2Bn, R1 = Me 13b3: R = -NHCH2-i-Pr, R1 = Me 13b4: R = -NH (CH2) 3-, R1 = Me

Scheme 3. Synthesis of compounds (11), (12) and (13). to HCI.H2NCH2CO2CH2CH3 or HCLH2NCH (Bn) CO2CH3 or

HCl.H2NCH (iPr) CO2 CH3 or HCl.HN (CH2) 3 CO2 CH3, NMM, CH2 Cl2, O0 C, 30 min., 40% (11a1), 55% (11a2), 50% (11a3), 84% (11a4) ), 83% (11b2), 78% (11b3), 95% (11b4). b Compound 5, EDC.HCl, NMB HOBt1, CH2 Cl2, 0.01C1h, then r, 12h, 40% (12a1), 40% (12a2), 42% (12a3), 50% (12a4), 50% ( 12b2), 75% (12b3), 51% (12b4). ç. MeOH, COCl K 2 CO 3, r.t. 30 min., 92% (13b2), 74% (13b3), 90% (13b4).

20

Purification and expression of HCV NS3 / 4A protease

Recombinant plasmid pET21d_6xHis-Tev-HCVNS3pro / 4A (cloned into Carlos Chagas Filho Institute of Biophysics Structural Genomics Laboratory -IBCCF / UFRJ by Dr. Emmerson CB da Costa) was inserted into E. coli strain BL21 [ADE3] (Novagen ) and expression of the recombinant 6xHis-Tev-HCVNS3pro / 4A protein was induced using 0.5mM isopropyl? -Dthiogalactopyranoside (IPTG). The cells were centrifuged and the bacterial pellet was lysed for 6xHis-Tev-HCVNS3pro / 4A purification using HisTRAP (GE-HeaIthcare) affinity column (Gui-Xin Du, Li-Hua 5 Hou1 Rong-Bin Guan, Yi- Gang Tong1 Hai-Tao Wang, Stablishment of a simple in vitro assay for hepatitis C virus NS3 serine protease based on recombinant substrate and single-chain protease, World Journal of Gastroenterology, 2002 8 (6): 1088-93), followed by excision of the NS3pro / 4A 6xHis-tev tail with the TEV protease enzyme (Joseph J., Savithri HS, Mutational analysis of the NI-protease from pepper vein banding potyvirus, Archives Virology, 2000, 145, 2493-2002). The purity of NS3Pro / 4A was evaluated on 15% SDS-Page gel, and protein quantification was performed using the Coomassie (Bradford) Protein Assay Kit (Pierce) following the manufacturer's protocol.

NS3pro / 4A recombinant protease activity and inhibition assay

HCV:

HCV proteolytic activity was evaluated with the SensoLyte®520 HCV Protease Assay Kit * Fluorimetric * (AnaSpec1CA1USA). This assay makes it possible to screen anti-HCV-NS3pro / 4A 20 inhibitors with greater efficiency and sensitivity, based on the FRET (fluorecence resonance energy transfer) phenomenon, in which the peptide derived from the NS4A / NS4B cleavage site is labeled. with 5-FAM / QXL ™ 520 fluorophore, with 5-FAM fluorescence suppressed by QXL ™ 520. The quantification of proteolytic activity occurs when the FRET peptide is cleaved into two 25 fragments by NS3pro / 4A, thus 5-FAM fluorescence can be monitored at 490nm / 520nm excitation / emission. The great advantage of using the 5-FAM-labeled peptide is that the fluorescence emission occurs at long wavelength, suffering less interference from the autofluorescence of the tested compounds.

30

Methodology used in the dosage of inhibitory capacity of the compounds described in this patent.

The lyophilized compounds were resuspended in DMSO (100%) at a concentration of 20mM. Subsequently, 100μΜ of the compounds were preincubated at 25 ° C for 10 min with 56ng of the enzyme HCV-NS3pro / 4A in a final volume of 50μΙ_ of the SensoLyte®520 HCV Protease Assay Kit * (AnaSpec1GA1USA) reaction buffer following the manufacturer's protocol. The reaction was initiated by the addition of the 5-FAM / QXL ™ 520 5 FRET Peptide substrate whose peptide cleavage was monitored in the SpectraMax M5 (Molecular Devices) plate reader at excitation / emission equal to 490nm / 520nm by measuring the fluorescence generated at every 30 seconds for 60 min.

The initial velocity (Vi) reaction kinetic parameters were calculated by linear or polynomial 3rd order regression using the SigmaPIot version 10.0 and Excel 2007 software, respectively, and the Vi values converted to residual activity (%).

The initial screening of 17 compounds against HCV NS3Pro / 4A proteolytic activity is shown in Table 2. We can see that compounds 12a3, 13b3, 11a1, 12a4 had better inhibitory activity for HCV1 NS3 / 4A protease and Compound 12a3 was the most active, with an inhibitory capacity of 63 ± 13%. Subsequently, the dose response of the most active compound (12a3) was compared with the standard inhibitor Ac-DE-Dif-ECha-C (AnaSpec, CA1 USA), where we can observe an IC50 of 76 ± 14μΜ (Figure 1) and 0, 8 ± 0.3μΜ, respectively. These data show the need for structural changes in the molecule aiming at optimizing activity against the protease site. These modifications to the structure can be accomplished using the 12a3 scaffold itself as a prototype molecule.

Table 2 - Inhibitory activity of 17 compounds against HCV NS3Pro / 4A protease in an in vitro reaction with SensoLyte® 520 HCV Protease Assay Kit (Anaspec, CA, USA). Listed compounds are in decreasing order of inhibition

Compound Percent inhibition at 100μΜ concentration (mean% ± SD) * 12a3 63 ± 13 13b3 # 54 11a1 49 ± 17 12a4 * 30 12b2 * 26 13b2 "24 12a1 20 + 17 11b2 * 11 12b3" 11 11a4 * 10 11b4 # 8 11b3 # 7 & NI ■ tf -Q CN 13b4 * NI 12a2 NI 11a3 NI 11a2 NI * standard deviation obtained with compounds tested in duplicate.

# compounds tested only once.

NI -not inhibited

The present invention will be described in detail through the following

examples given below. It should be noted that the invention is not limited to these examples and includes variations and modifications within the limits to which it applies.

Example 1: Obtaining 1,4: 3,6-dianhydro-mono (4-methylbenzenesulfonate) -D-mannitol (2).

To a solution of isomanide (1g, 6mmol) and pyridine (1.1 OmL) in CH 2 Cl 2 (10mL) was added p-toluenesulfonyl chloride (1.49g, 7mmol) to

O0C. The reaction was stirred at room temperature for 12h and then diluted with CH 2 Cl 2 (20mL). The solution was washed with water, 1N HCl and saturated sodium chloride solution. After evaporation of the solvent, the solid was recrystallized from a methanol / isopropanol mixture providing the product as a white solid in 40% yield. MP: 103-104 ° C.

1 H NMR δ ppm (CDCl 3, 300MHz): 7.84 (d, 2H, J = 8.1 Hz); 7.34 (d, 2H, J = 8.1 Hz); 4.91 (dd, 1H, J = 6.6 / 5.5 Hz); 4.42 (t, 1H, J = 5.1 Hz); 4.49 (t, 1H, J = 4.8 Hz); 4.29-4.25 (m, 1H); 4.04-3.95 (m, 2H); 3.79 (t, 1H, J = 7.8 Hz); 3.55 (dd, 1H, J = 7.2 / 1.8 Hz); 2.46 (s, 3Η). IRv CrrT1 (KBr): 3526, 2933, 2866, 1596, 1359, 1189, 1173, 1050, 1019, 818, 663.

Example 2: Obtaining 1,4: 3,6-dianhydro-5-0- (benzyl) -2- (4-methylbenzenesulfonate) -D-mannitol (3).

To a solution of 2 (10g, 33mmol), 50% aqueous KOH and TBAB (0.322g, 1mmol) in CH 2 Cl 2 (80mL) was added benzyl chloride (6.54g, 38mmol). After stirring for 12h, the mixture was diluted with water (10mL) and the organic phase separated. The aqueous phase was extracted with CH 2 Cl 2 and dried with Na 2 SO 4. The product was purified by silica gel column chromatography eluting with a mixture of hexane and ethyl acetate, providing the product as a colorless oil in 75% yield.

1 H NMR δ ppm (CDCl 3, 300MHz): 7.81 (d, 2H, J = 8.4 Hz); 7.35-7.32 (m, 7H); 4.91-4.89 (m, 1H); 4.69 (d, 1H, J = 12.0 Hz); 4.50 (d, 1H, J = 12.0 Hz); 4.47 (t, 2H, J = 2.1 Hz); 4.04-3.79 (m, 4H); 3.62 (t, 1H, J = 8.4 Hz); 2.44 (s, 3H). IR v cm -1 (film): 3063, 2977, 2950, 2879, 1598, 1454, 1366, 1190, 1178, 1141, 1027, 853. Ao 20 = +98 (c, 0.1) DMSO.

Example 3: Obtaining 1,4: 3,6-dianhydro-2-azido-2-deoxy-5-0- (benzyl) -DGlucitol (4).

A mixture of 3 (1.86g, 4mmol), [bmin] + [BF4] 'previously dried under vacuum at 90 ° C (4.8mL, 23mmol) and NaN3 (0.93g, 14mmol) was heated to 120 ° C by 12pm. Water was added and the aqueous phase was extracted with ethyl ether. The product was purified by flash column chromatography (hexane / ethyl acetate) to give a light yellow oil in 73% yield.

1 H NMR δ ppm (CDCl 3, 300MHz): 7.36-7.26 (m, 5H); 4.75 (d, 1H, J = 11.7 Hz); 4.66 (t, 1H, J = 4.5 Hz); 4.54 (d, 1H, J = 11.7 Hz); 4.48 (d, 1H1 J = 4.2 Hz); 4.15-4.00 (m, 4H); 3.86 (dd, 1H, J = 6.3 / 2.7 Hz); 3.66 (t, 1H, J = 7.8 Hz). 13 C NMR δ ppm (CDCl 3, 75.4 MHz): 137.4 (-C), 128.3 (-CH), 127.8 (-CH), 127.7 (-CH), 86.4 (- CH), 80.5 (-CH), 78.8 (-CH), 72.6 (-CH2), 72.4 (-CH2), 70.7 (-CH2), 66.2 (-CH) . IRvcm-1 (film): 3063, 3031, 2946, 2878, 2102, 1455, 1320, 1256, 1135, 1100, 1083, 1021, 739. HRMS calculated for C13 H15 N3 O5 Na, 284.1011; found, 284.1025. +92 (c, 0.1) DMSO.

Example 4: Obtaining 1,4: 3,6-dianhydro-2-amino-2-deoxy-5-0- (benzyl) -DGlucitol (5)

To a solution of 4 (0.870g, 3mmol) in ethanol (25mL) was added 5% Pd / C (0.3mmol) and the mixture was hydrogenated under 40psi for 5h. After this time the suspension was filtered on XAD-4 and washed with ethanol. Evaporating the solvent gave a colorless oil in quantitative yield.

1 H NMR δ ppm (CDCl 3, 300MHz): 7.37-7.35 (m, 5H); 4.76 (d, 1H, J = 11.4 Hz); 4.68 (t, 1H, J = 4.2 Hz); 4.54 (d, 1H, J = 12.0 Hz); 4.27 (d, 1H, J = 4.2 Hz); 4.06 - 4.02 (m, 1H); 3.85 (dd, 1H, J = 6.3 / 2.4 Hz); 3.76 (d, 1H, J = 9.3 Hz); 3.64 (t, 1H, J = 7.8 Hz); 3.53 (d, 1H, J = 4.2 Hz); 3.45 (s, 1H). 13 C NMR δ ppm (CDCl 3, 75.4 15 MHz): 137.9 (-C), 128.6 (-CH), 128.0 (-CH), 127.8 (-CH), 90.0 ( -CH), 80.1 (CH), 79.4 (-CH), 76.3 (-CH2), 72.6 (-CH2), 70.5 (-CH2), 58.8 (-CΗ) . IR v cm '

1 (film): 3360, 2876, 1605, 1455, 1369, 1209, 1065, 1017, 751, 700. HRMS calculated for C 13 H 17 NO 3 Na, 236.1287; found, 236.1284.

Example 5: Obtaining Ionic Liquid [Bmim] + BF4 '(compound 9).

To a solution of N-methyl imidazole (200mmol, 16mL) in toluene (25mL) at 0 ° C was added 1-Bromobutane (230mmol, 25mL) slowly and with stirring. The mixture was refluxed (1100 ° C) for 36 h. Two phases were formed: the densest (viscous) and the lightest. They were separated. The viscous phase was washed with ethyl ether (3 x 50mL) and ethyl acetate (3x50mL). Soon after, CH 2 Cl 2 (150mL) and NaBF 4 (164mmol, 18g) were added. The mixture was stirred for 24h at room temperature. After this time, the mixture was filtered 2 times, the activated charcoal added and left for an additional 24h under stirring. At the end, the mixture was filtered, evaporated and finally filtered on neutral alumina by washing well with CH 2 Cl 2. The product was obtained as a light yellow oil in 83% yield. 1 H NMR δ ppm (CDCl 3, 300 MHz): 9.27 (s, 1H); 7.38 (s, 1H); 7.32 (s, 1H); 4.22 (t, 2Η, J = 7.2 Hz); 3.99 (s, 3H); 1.91-1.80 (m, 2H); 1.43-1.30 (m, 2H); 0.94 (t, 3Η, J = 7.2 Hz). IR VcrrT1 (KBr): 3606, 3418, 3161, 3119, 2964, 2939, 2877, 1634, 1574, 1467, 1385, 1338, 1286, 1170, 1062, 848, 754, 623.

5th

General process for obtaining acids 11a-b.

To a solution of anhydride 10a-b (4.02mmol, 0.868g) in anhydrous dichloromethane (5 ml) at 0 ° C was added dropwise a solution of the appropriate amino ester hydrochloride (4.82 mmol) in anhydrous dichloromethane ( 2.5mL) containing N-methyl morpholine (4.82mmol, 0.53mL). The mixture was stirred for 12h and the solvent evaporated after that time. The residue was dissolved in ethyl acetate (or dichloromethane) and washed successively with 10% HCl and saturated NaCl solution. The organic phases were dried over NaaSO4 and concentrated to yield products 11a1-a3 and 11b2-b3 as viscous oils which were purified by recrystallization (CH2 Cl2 / MeOH) to obtain a 40% (11a1), 55% (11a2) hygroscopic solid. , 50% (11a3), 84% (11b2) and 78% (11b3) yield. The free amine of L-proline methyl ester was obtained by dissolving the salt (7.87 mmol, 1.3 g) in a minimum volume of 20% K2 CO3 and extracted eight times with ethyl ether. Concentration of the organic phases provided the free amine (4.02mmol, 0.519g) as an oil which was directly dissolved in dichloromethane (2.5mL) and added to a solution of anhydride 10 (4.02mmol, 0.868g) in anhydrous dichloromethane. (5mL). The mixture was stirred for 12h and the solvent was evaporated. The product was purified by recrystallization (CH 2 Cl 2 / MeOH) to obtain a hygroscopic solid (11a4) in 84% and (11b4) in 95% yield.

Example 6: Obtaining (2S, 3S) -2,3-diacetoxy-4- (2-ethoxy-2-oxoethylamino) -4-oxobutanoic acid (11a1). 1 H NMR (CDCl 3, 300 MHz): 5.83 (d, 1H, J = 2.4 Hz), 5.64 (d, 1H, J = 2.4 Hz), 4.23 (q, 2H, J = 7.2 Hz), 4.13 (1H, dd, J = 18.3, 5.1 Hz), 3.99 (dd, 1H, J = 18.3, 5.1 Hz), 2.22 (s, 3H), 2.17 (s, 3H), 1.30 (t, 3H, J = 7.2 Hz). 13 C-NMR (CDCl 3, 75 MHz): 169.8, 169.6, 169.3, 169.1,

166.2, 71.7, 70.7, 61.8, 41.2, 20.4, 20.2, 14.0. IR (film) ν cm -1: 3851, 3626, 3369, 2926, 2853, 2479, 1746, 1731, 1694, 1681, 1652, 1634, 1538, 1532, 1463, 1455, 1377, 1271, 1212, 1116, 1073 , 1032, 992, 896, 866, 799, 738, 702. [α] D 20 + 20 ° (c 0.10, CH 2 Cl 2). HRMS (FAB): Calculated for C 12 H 17 NO 9 [M + 1]: 319.0903. Found: 320.0611.

Example 7: Obtaining (2S, 3S) -2,3-diacetoxy-4 - ((S) -1-methoxy-1-oxo3-phenylpropan-2-ylamino) -4-oxobutanoic acid (11a2). 1 H NMR δ ppm (CDCl3, 300 MHz): 7.32-7.18 (m, 5H), 7.06 (d, 1H, J = 5.4 Hz), 5.74 (sl, 1H), 5.34 (sl, 1H), 4.79 (dd, 1H, J = 8.4, 3.9 Hz), 3.66 (s, 3H), 3.07 (m, 2H), 2.05 (s, 3H), 2.04 (s, 3H). 13 C NMR δ ppm (50 MHz, CDCl 3): 171.9, 171.2, 170.2, 169.4, 166.4, 135.2, 10 129.1, 128.6, 127.2, 72.3, 70.7, 52.6, 52.3, 37.3, 20.4, 20.3. IR (KBr) ν cm -1: 3627, 3585, 3389, 2955, 1732, 1716, 1681, 1652, 1538, 1497, 1455, 1436,

1374, 1220, 1132, 1069, 946, 877, 738, 702. [α] D20 +26.2 (c 1.0, CH 2 Cl 2). HRMS calcd for C13H2INOg [M + 1]: 395.1216. Found: 396.6837.

Example 8: Obtaining (2S, 3S) -2,3-diacetoxy-4- (S) -1-methoxy-3-methyl-1-oxobutan-2-ylamino) -4-oxobutanoic acid (11a3). 1 H NMR δ ppm (CDCl 3, 300 MHz): 6.68 (d, 1H, J = 7.6 Hz), 5.76 (d, 1H, J = 2.4 Hz), 5.64 (d, 1H, J = 2.7 Hz), 4.13 (dd , 1H, J = 14.4, 7.2 Hz), 3.76 (s, 3H), 2.22 (s, 3H), 2.17 (s, 3H), 1.28-1.24 (m, 1H), 0.91 (d, 3H, J = 6.9 Hz), 0.86 (d, 3H, J = 6.9 Hz). 13 C NMR δ 20 ppm (50 MHz, CDCl 3): 172.0, 169.8, 169.6, 169.2, 166.1, 72.1, 70.5, 56.9, 52.3,

31.0, 20.4, 20.1, 18.8, 17.4. IRv (cm -1) KBr: 3854, 3752, 3736, 3650, 3399, 2966, 2936, 2850, 2615, 1737, 1662, 1540, 1439, 1376, 1221, 1151, 1129, 1073, 1017, 945, 892, 823. [a] D 20 +12.6 (c 1.0, CH 2 Cl 2). HRMS calcd for C 14 H 21 NO 9 [M + 1]: 347.1216. Found: 348.6792.

25

Example 9: Obtaining (2S, 3S) -2,3-diacetoxy-4- (2- (methoxycarbonyl) pyrrolidin-1-yl) -4-oxobutanoic acid (11a4). 1H NMR δ ppm

(CDCl3, 300 MHz): 5.74 (d, 1H, J = 4.5 Hz), 5.59 (d, 1H, J = 4.5 Hz), 4.43 (dd, 1H, J = 8.2, 3.0 Hz), 4.00-3.94 (m 1H), 3.69 (s, 3H), 3.60-3.51 (m, 1H), 2.16 (s, 3H), 2.15 (s, 3H), 2.11-2.09 (m, 2H), 2.07-1.98 (m, 2H) . 13 C-NMR δ ppm (50 MHz, CDCl 3): 171.8, 169.9, 169.7, 169.1, 165.1, 70.9, 70.3, 59.7, 52.2, 46.9,

28.5, 24.7, 20.4, 20.3. IR ν (cm -1) KBr: 3315, 2965, 2882, 1753, 1669, 1538, 1453, 1368, 1212, 1136, 1096, 1078, 961, 881, 740. [a] D20-47.0 (c1.0, CH 2 Cl 2). HRMS calcd for C 14 H 19 NO 9 [M + 1]: 345.1060. Found: 346.1280.

Example 10: Obtaining (2R, 3R) -2,3-diacetoxy-4 - ((S) -1-methoxy-1-oxo-3-phenylpropan-2-ylamino) -4-oxobutanoic acid (11b2). 1H NMR δ ppm

(CDCl3, 300 MHz): 7.32-7.22 (m, 5H), 6.60 (d, 1H, J = 7.9 Hz), 5.73 (br s, 1H), 5.50 (br s, 1H), 4.88 (q, 1H, J = 5.7 Hz), 3.73 (s, 3H), 3.15-3.06 (m, 2H), 2.07 (s, 3H), 2.05 (s, 3H). 13 C-NMR δ ppm (50 MHz, CDCl 3): 171.2, 171.1, 169.9, 169.1,

166.1, 135.2, 129.1, 128.6, 127.3, 72.3, 70.6, 52.6, 52.4, 37.4, 20.4, 20.3. IRv (cm -1) KBr: 3500, 2957, 1750, 1677, 1534, 1455, 1374, 1214, 1115, 1063, 972, 703. [α] D 20 + 32.5 (c 0.8, CH 2 Cl 2) HRMS calculated for C 18 H 21 NO 9 [ M + 1]: 395.1216 Found: 396.1488.

Example 11: Obtaining (2α, 3α) -2,3-diacetoxy-4 - ((S) -1-methoxy-3-15-methyl-1-oxobutan-2-ylamino) -4-oxobutanoic acid (11b3). 1 H NMR δ ppm (CDCl 3, 300 MHz): 6.77 (d, 1H, J = 8.9 Hz), 6.76 (br s, 1H), 5.73 (d, 1H, J = 2.4 Hz), 5.58 (d, 1H, J = 2.4 Hz), 4.53 (dd, 1H, J = 8.8, 4.8 Hz), 3.74 (s, 3H), 2.20 (s, 3H), 2.15 (s, 1H), 2.14 (s, 1H), 0.90 (d , 3H, J = 6.8 Hz), 0.86 (d, 3H, J = 6.8 Hz). 13 C-NMR (50 MHz, CDCl 3): 172.0, 170.2, 169.9, 169.4, 166.5, 72.4, 71.0, 56.9, 20 52.3, 31.1, 20.4, 20.1, 18.9, 17.5. IR v (cm -1) movie: 3344, 2966, 1745, 1535, 1438, 1375, 1214, 1151, 1120, 1066, 973, 875, 736. [α] D 20 + 26.7 (c 0.9, CH 2 Cl 2). for C 14 H 21 NO 9 [M + 1]: 347.1216 Found: 348.1465.

Example 12: Obtaining (2R, 3α) -2,3-diacetoxy-4 - ((S) - (2- (methoxycarbonyl) pyrrolidin-1-yl) -4-oxobutanoic acid (11b4) 1H NMR δ ppm

(CDCl3, 300 MHz): 5.74 (d, 1H, J = 4.3 Hz), 5.63 (d, 1H, J = 4.3 Hz), 4.44 (dd, 1H, J = 8.4, 3.1 Hz),), 4.00-3.95 (m 1H), 3.69 (s, 3H), 3.57-3.52 (m, 1H), 2.17 (s, 3H), 2.16 (s, 3H), 2.11-2.07 (m, 2H), 2.04-1.95 (m, 2H). 13 C-NMR (50 MHz, CDCl 3): 171.8, 169.9, 169.7, 168.9, 70.7, 69.9, 59.7, 52.3, 47.0, 28.6, 24.7,

20.4, 20.3. IRv (cm -1) movie: 3449, 2957, 1751, 1655, 1637, 1618, 1459, 1438,

1375, 1220, 1120, 1073, 874, 733. [α] D 20 -42.5 (c 0.8, CH 2 Cl 2). HRMS calcd for C 14 H 19 NO 9 [M + 1]: 345.1060. Found: 346.1271. General process for obtaining compounds 12a-b.

To a solution of the corresponding acid 11a-b (0.515mmol) and amine 5 (0.618mmol, 0.145g) in anhydrous dichloromethane (10mL) at 0 ° C under inert atmosphere, EDC.HCI (0.790mmol, 0.125g) was added. ), HOBt (0.790mmol, 0.106g) and NMM (1.614mmol, 0.17mL). The mixture was stirred at 0 ° C for 1h and at room temperature for 12h. The solvent was evaporated and the residue dissolved in ethyl acetate and washed successively with saturated NaHCO 3 solution, 10% HCl and saturated NaCl solution. After drying over Na 2 SO 4 and concentrated, crude product 12a-b was obtained.

Example 13: Obtaining (2S, 3S) -1 - ((3S, 6f?) - 6- (benzyloxy) hexahydrofuro [3,2-

b] furan-3-ylamino) -4- (2-ethoxy-2-oxoethylamino) -1,4-dioxobutane-2,3-dil diacetate (12a1). 1 H NMR δ ppm (CDCl 3, 300 MHz): 77.37-7.35 (m, 5H), 6.75 δ (t, 1H, J = 4.8 Hz); 6.42 (d, 1H, J = 8.7 Hz), 5.65 (d, 1H, J = 3.6 Hz), 5.53 (d, 1H, J = 3.3 Hz), 4.76 (d, 1 H1J = 11.7 Hz), 4.62 (t , 1H, J = 4.8 Hz), 4.55 (d, 1H, J = 11.7 Hz), 4.42 (d, 1H, J = 4.2 Hz), 4.39-4.34 (m, 1H), 4.22 (q, 2H, J = 7.2 Hz), 4.10-4.05 (m, 4H), 4.00 (d, 1H, J = 5.1 Hz) 3.86 (dd, 1H, J = 9.0, 7.5 Hz), 3.67 (dd, 1H, J = 9.0, 7.5 Hz ), 2.18 (s, 3H), 2.17 (s, 3H), 1.29 (t, 3H, J = 7.2 Hz). 13 C NMR (50 MHz, CDCl 3): 169.3, 169.0, 168.9, 166.3, 165.7, 137.6, 128.4,

127.8, 127.7, 86.6, 80.3, 78.9, 72.9, 72.5, 71.2, 71.9, 70.8, 61.7, 57.0, 41.1,

20.5, 20.4, 14.0. IRv (cm -1) movie: 3311, 3066, 3032, 2959, 2929, 2875, 1746, 1669, 1602, 1546, 1496, 1455, 1374, 1344, 1274, 1225, 1134, 1094, 1085, 1070, 1017, 962, 912, 880, 845, 798, 780, 746, 698, 662, 638, 603. [α] D 20 + 28.0 ° (c 0.10, CH 2 Cl 2). Appearance: yellow oil. Yield: 40% after purification by column chromatography (eluent: hexane / ethyl acetate). HRMS calcd for C 25 H 32 N 2 O 11 [M + 1]: 536.5284; Found 537.73676.

Example 14: Obtaining (2S, 3S) -1 - ((3S, 6R) -6- (benzyloxy) hexahydrofuro [3,2-b] furan-3-ylamino) -4 - ((S) -1-methoxy -1-oxo-3-phenylpropan-2-ylamino) -1,4-

dioxobutane-2,3-dil diacetate (12a2). 1 H NMR δ ppm (CDCl3, 300 MHz): 7.35-7.26 (m, 8H), 7.05 (d, 2H, J = 4.2 Hz), 6.55 (d, 1H, J = 4.8 Hz), 6.46 (d, 1H, J = 4.5 Hz), 5.66 (d, 1H, J = 1.5 Hz), 5.54 (d, 1H, J = 1.5 Hz), 5.30 (s, 2H), 4.62 (dt, 1H, J = 11.4, 2.7 Hz) , 4.55 (d, 1H, J = 6.9 Hz), 4.41 (dd, 1H, J = 10.1, 2.4 Hz), 4.35-4.33 (m, 1Η), 4.08-4.03 (m, 2Η), 3.85-3.82 (m , 2Η), 3.75 (s, 3Η), 3.66 (t, 1Η, J = 5.0 Hz), 3.13 (dd, 1H, J = 11.6, 8.1 Hz), 3.08 (dd, 1H, J = 11.6, 8.1 Hz) ,

2.13 (s, 3H), 2.06 (s, 3H). 13 C-NMR (50 MHz, CDCl 3): 169.3, 169.2, 168.7,

165.8, 165.7, 137.6, 135.2, 129.2, 128.7, 128.5, 127.9, 127.8, 127.4, 86.8, 86.7, 80.3, 79.0, 73.0, 72.5, 72.3, 70.8, 57.1, 52.6, 52.4, 37.5, 20.5, 20.4. IR ν (cm -1) KBr: 3286, 3064, 3031, 2952, 2880, 2359, 1755, 1657, 1540, 1497, 1455, 1436, 1373, 1343, 1374, 1256, 1206, 1136, 1098, 1084, 1058 , 1029, 963, 912, 879, 749, 700. [α] D 20 + 50.0 (c 0.10, CH 2 Cl 2). Appearance: yellow solid (mp 122-123 ° C). Yield: 40% after recrystallization (dichloromethane / ethyl ether). HRMS calcd for C 31 H 36 N 2 O 11 [M + 1]: 612.2319; Found 613.7522.

Example 15: Obtaining (2S, 3S) -1 - ((3S, 6%) - 6- (benzyloxy) hexahydrofuro [3,2- b] furan-3-ylamino) -4 - ((S) -1 -methoxy-3-methyl-1-oxobutan-2-ylamino) -1,4-dioxobutane-2,3-dil diacetate (12a3). 1 H NMR δ ppm (CDCl 3, 300 MHz): 7.35-15 7.33 (m, 5H), 6.68 (d, 1H, J = 5.1 Hz), 6.58 (d, 1H J = 4.5 Hz), 5.66 (d, 1H, J = 2.1 Hz), 5.56 (d, 1H, J = 1.8 Hz), 5.51 (s, 2H), 4.74 (d, 1H, J = 7.2 Hz), 4.64 (t, 1H, J = 3.0 Hz), 4.55 (t, 1H, J = 3.0 Hz), 4.35-4.33 (m, 1H), 4.06-4.03 (m, 2H), 3.87-3.80 (m, 3H), 3.73 (s, 3H), 2.17-2.15 (m , 1H), 2.17 (s, 3H), 2.16 (s, 3H),

0.91 (d, 3H, J = 3.9 Hz), 0.87 (d, 3H, J = 4.2 Hz). 13 C-NMR (50 MHz, CDCl 3): 169.3, 169.2, 168.9, 166.2, 166.0, 137.6, 128.3, 127.8, 127.7, 86.6, 86.5, 80.2,

78.9, 72.7, 72.4, 72.1, 70.8, 57.0, 56.9, 52.2, 30.9, 20.5, 20.4, 18.7. IR ν (cm -1) KBr: 3851, 3742, 3646, 3287, 3065, 2962, 2879, 1754, 1656, 1539, 1455, 1436, 1373, 1274, 1207, 1141, 1098, 1059, 1027, 964, 876 , 845, 750, 699, 651. [α] D20 +24.0 (c0.10, CH2 Cl2). Appearance: yellow solid (mp 112-113 ° C). Yield: 25% after recrystallization (dichloromethane / ethyl ether). HRMS calcd for C27H36N2O11 [M + 1]: 564.5815; Found 565.76803.

Example 16: Obtaining (2S, 3S) -1 - ((3S, 6%) - 6- (benzyloxy) hexahydrofuro [3,2- b] furan-3-ylamino) -4 - (%) - ( 2- (methoxycarbonyl) pyrrolidin-1-yl) -1,4-dioxobutane30 2,3-dil diacetate (12a4). 1 H NMR δ ppm (CDCl 3, 300 MHz): 7.36-7.28 (m, 5H), 6.48 (br s, 1H), 5.68 (d, 1H, J = 4.9 Hz), 5.55 (d, 1H, J = 4.9 Hz ), 4.78 (d, 1H, J = 11.7 Hz), 4.64-4.61 (m, 1H), 4.58 (d, 1H, J = 11.7 Hz), 4.43-4.32 (m, 3H), 4.12-4.03 (m, 2H), 3.96-3.90 (m, 1H), 3.90-3.83 (m, 2H), 3.78 (s, 1H), 3.69 (s, 3H), 2.21-2.18 (m, 2H), 2.15 (s, 3H) 2.10 (s, 3H), 2.08-2.00 (m, 2H). 13 C-NMR (50 MHz, CDCl 3): 171.7, 169.5, 169.1, 165.7, 164.7, 137.6, 128.4, 127.9, 127.8,

86.7, 80.3, 78.9, 72.8, 72.5, 71.2, 70.9, 70.3, 59.3, 57.0, 52.1, 46.8, 28.8, 24.6,

20.6, 20.3. IR ν (cm -1) KBr: 3414, 2953, 2882, 1751, 1665, 1535, 1451, 1438, 1372, 1213, 1135, 1071, 960, 700. [α] D20 +13.0 (c 0.8, CH2 Cl2). Appearance: hygroscopic white solid. Yield: 50% after purification by column chromatography (eluent: hexane / ethyl acetate). HRMS calcd for C27H34N2O1 [M + 1]: 562.2163; Found 563.2430.

Example 17: Obtaining (2 /?, 3 /?) -1 - ((3S, 6 /?) - 6- (benzyloxy) hexahydrofuro [3,2-6,6] furan-3-ylamino) -4- ( (S) - (1-Methoxy-1-oxo-3-phenylpropan-2-ylamino) -1,4-

dioxobutane-2,3-dil diacetate (12b2). 1 H NMR δ ppm (CDCl 3, 300 MHz):

7.35-7.24 (m, 8H), 7.04 (d, 2H, J = 6.2 Hz), 5.66 (d, 1H, J = 2.8 Hz), 5.54 (d, 1H, J = 2.8 Hz), 4.87 (dd, 1H , J = 13.7, 5.5 Hz), 4.73 (d, 1H, J = 11.8 Hz), 4.61 (t, 1H, J = 4.6 Hz), 4.53 (d, 1H, J = 11.8 Hz), 4.40 (d, 1H , J = 4.3 Hz), 4.34-4.32 (m, 1H), 4.06-4.02 (m, 2H), 3.85-3.82 (m, 2H), 3.74 (s, 3H), 3.67 (dd, 1H, J =

9.0, 7.3 Hz), 3.15 (dd, 1H, J = 13.9, 5.4 Hz), 3.08 (dd, 1H, J = 13.9, 5.4 Hz), 2.12 (s, 3H), 2.05 (s, 3H). 13 C-NMR (50 MHz, CDCl 3): 171.2, 169.3, 168.7,

165.9, 165.7, 137.6, 135.2, 129.2, 128.6, 128.4, 127.9, 127.8, 127.3, 86.7, 80.3,

78.9, 72.9, 72.5, 72.2,, 71.8, 70.8, 57.1, 52.6, 52.4, 37.5, 20.5, 20.4. IR ν (cm -1) KBr: 3287, 3066, 3030, 2953, 2879, 1755, 1657, 1540, 1497, 1436, 1372, 1257,

1204, 1134, 1083, 1057, 962, 747, 699. [α] D 20 +68.2 (c 0.8, CH 2 Cl 2). Appearance: white solid (mp 178-180 ° C). Yield: 50% after purification by column chromatography (eluent: hexane / ethyl acetate). HRMS calcd for C31H36N2O11 [M + 1]: 612.2319; Found 613.2689.

25

Example 18: Obtaining (2 /?, 3 /?) -1 - ((3S, 6 /?) - 6- (benzyloxy) hexahydrofuro [3,2- Î ±] furan-3-ylamino) -4 - (( S) -1-Methoxy-3-methyl-1-oxobutan-2-ylamino) -1,4-dioxobutane-2,3-dil diacetate (12b3). 1 H NMR δ ppm (CDCl 3, 300 MHz): 7.34-7.27 (m, 5H), 6.70 (d, 1H, J = 8.7 Hz), 6.61 (d, 1H J = 7.4 Hz), 5.66 (d, 1H, 30 J = 3.1 Hz), 5.56 (d, 1H, J = 3.1 Hz), 4.75 (d, 1H, J = 11.8 Hz), 4.64-4.61 (m, 1H), 4.54 (d, 1H, J = 11.8 Hz) , 4.51 (dd, 1H, J = 8.8, 4.8 Hz), 4.42-4.40 (m, 1H), 4.30-4.33 (m, 1H), 4.06-4.03 (m, 2H), 3.87-3.82 (m, 2H) , 3.73 (s, 3H), 3.65 (dd, 1H, J = 9.0, 7.3 Hz), 2.17 (s, 3H), 2.15 (s, 3H), 2.13 (s, 1H), 0.89 (d, 3H, J = 6.8 Hz), 0.85 (d, 3H, J = 6.8 Hz). 13 C-NMR (50 MHz, CDCl 3): 171.7, 169.4, 169.0, 166.2, 165.8, 137.6, 128.4, 127.9, 127.8, 86.7, 80.3, 78.9, 72.9, 72.5, 72.4,

71.8, 70.8, 57.1, 56.9, 52.2, 31.0, 20.6, 20.5, 18.2, 17.5. IRv (cm -1) KBr: 3279, 2963, 2876, 1753, 1654, 1541, 1468, 1373, 1259, 1208, 1144, 1061, 963, 874, 737. [α] δ 20 +71.2 (c 0.8, CH 2 Cl 2) . Appearance: white solid (mp 84-85 ° C). Yield: 75% after chromatographic column purification (eluent: hexane / ethyl acetate). HRMS calcd for C27H36N2O11 [M + 1]: 564.2319; Found 565.2684.

Example 19: Obtaining (2R, 3 /?) -1 - ((3S, 6 /?) - 6- (benzyloxy) hexahydrofuro [3,2-10 fo] furan-3-ylamino) -4 - ((R ) -2- (methoxycarbonyl) pyrrolidin-1-yl) -1,4-dioxobutane2,3-dil diacetate (12b4). 1H NMR δ ppm (CDCl3, 300 MHz): 7.36-7.28 (m, 5H), 6.61 (d, 1H1 J = 7.5 Hz), 5.68 (d, 1H, J = 5.0 Hz), 5.54 (d, 1H, J = 5.0 Hz), 4.74 (d, 1H, J = 11.8 Hz), 4.64 (t, 1H, J = 4.7 Hz), 4.57 (d, 1H, J = 11.8 Hz), 4.42-4.40 (m, 1H), 4.35-4.33 (m, 2H), 4.08-4.03 (m, 3H), 3.96-3.91 (m, 1H), 3.84-15 3.84 (m, 3H), 3.69 (s, 3H), 2.23-2.19 (m, 2H), 2.15 (s, 3H), 2.10 (s, 3H), 2.04-1.97 (m, 2H). 13 C-NMR (50 MHz, CDCl 3): 171.7, 169.5, 169.2, 165.8, 164.7,

137.6, 128.4, 127.9, 127.8, 86.7, 80.3, 78.9, 72.9, 72.5, 71.2, 70.9, 70.3, 59.3,

57.1, 52.2, 46.8, 28.8, 24.6, 20.7, 20.3. IR ν (cm -1) KBr: 3314, 2955, 2880, 1751, 1668, 1538, 1453, 1372, 1212, 1135, 1096, 1070, 961, 881, 742. [α] D 20 + 20.5 (c 0.8, CH 2 Cl 2 ). Appearance: white solid (mp 53-54 ° C). Yield: 51% after purification by column chromatography (eluent: hexane / ethyl acetate). HRMS calcd for C27H34N2O11 [M + 1]: 562.2163; Found 563.2421.

Example 20: Obtaining (2S) -methyl-2 - ((2R, 3R) -4 - ((3S, 6%) - 6- (benzyloxy) hexahydrofuro [3,2-b] furan-3-ylamino) -2,3-dihydroxy-4-

oxobutanamido) -3-phenylpropanoate (13b2). 1 H NMR δ ppm (CD 3 OD, 300 MHz): 7.37-7.18 (m, 10H), 4.76-4.75 (m, 1H), 4.71 (d, 1H, J = 11.4 Hz), 4.70 (t, 1H, J = 4.7 Hz), 4.53 (d, 1H, J = 11.4 Hz), 4.44 (d, 1H, J = 4.4 Hz), 4.41 (d, 1H, J = 1.8 Hz), 4.39 (d, 1H, J = 1.8 Hz) 4.34-4.31 (m, 1H), 4.13 (dd, 1H, J = 11.6, 30 6.6 Hz), 4.04 (dd, 1H, J = 9.6, 4.8 Hz), 3.87-3.85 (m, 1H), 3.84- 3.82 (m, 1H), 3.70 (s, 3H), 3.64 (dd, 1H, J = 9.0, 7.0 Hz), 3.18 (dd, 1H, J = 13.8, 5.6 Hz), 3.08 (dd, 1H, J = 13.8, 5.6 Hz). NMR13C (50 MHz, CD3OD): 174.3, 174.0, 173.1, 139.4, 137.6, 130.4, 129.6, 129.4, 129.1, 128.9, 128.0, 88.5, 81.5, 80.7, 74.1, 74.0, 73.5, 71.8, 71.7, 58.2, 54.8, 52.8, 38.6. IR ν (cm -1) KBr: 3393, 30663, 3030, 2950, 2879, 1736, 1670, 1648, 1541, 1497, 1454, 1368, 1215, 1139, 1081, 911, 737, 700. [a] D20 + 124.3 (c 0.6, MeOH). Appearance: yellow solid (Rf. 41-42 ° C). Yield: 92% after purification by chromatographic column 5 (eluent: hexane / ethyl acetate). HRMS calcd for C27H32N2O9 [M + 1]: 528.2108; Found 529.2430.

Example 21: Obtaining (2S) -methyl-2 - ((2R, 3R) -4 - ((3S, 6%) - 6- (benzyloxy) hexahydrofuro [3,2b] furan-3-ylamino) -2 1,3-dihydroxy-4-oxo-butanamido) -3-methyl butanoate (13b3). 1 H NMR δ ppm (CD 3 OD, 300 MHz): 7.38-7.26 (m, 5H), 4.70-4.69 (m, 2H), 4.54 (d, 1H, J = 11.5 Hz), 4.49 (d, 1H J = 1.7 Hz ), 4.47 (d, 1H, J = 4.4 Hz), 4.43 (d, 1H, J = 1.8 Hz), 4.41 (d, 1H, J = 5.6 Hz), 4.34-4.32 (m, 1H), 4.14 (dd 1H, J = 11.6, 6.6 Hz), 4.05 (dd, 1H, J =

9.6, 4.8 Hz), 3.89-3.86 (m, 1H), 3.85-3.82 (m, 1H), 3.73 (s, 3H), 3.55 (dd, 1H, J = 9.0, 7.0 Hz), 2.21-2.14 (m , 1H), 0.96 (d, 3H, J = 4.0 Hz), 0.94 (d, 3H, J = 4.0 Hz). 13 C-NMR (50 MHz, CD 3 OD): 174.3, 174.0, 173.2, 139.4, 129.4, 129.1,

128.9, 88.5, 81.8, 80.7, 74.1, 74.0, 73.9, 73.5, 71.8, 58.7, 58.2, 52.7, 32.4, 19.3, 18.3. IR ν (cm -1) KBr: 3387, 2961, 2877, 1751, 1655, 1638, 1541, 1438, 1371, 1272, 1210, 1141, 1023, 735, 699. [α] D 20 + 96.0 (c 0.9, MeOH ). Appearance: yellow solid 20 (mp 44-45 ° C). Yield: 74% after chromatographic column purification (eluent: hexane / ethyl acetate). HRMS calcd for C 23 H 32 N 2 O 9 [M + 1]: 480.2108; Found 481.2320.

Example 22: Obtaining (2R) -methyl-1 - ((2R, 3R) -4 - ((3S, 6%) - 6- (benzyloxy) hexahydrofuro [3,2b] furan-3-ylamino) -2 , 3-dihydroxy-4-

oxobutanoyl) pyrrolidine-2-carboxylate (13b4). 1 H NMR δ ppm (CD 3 OD, 300 MHz): 7.36-7.28 (m, 5H), 4.74 (d, 1H, J = 11.7 Hz), 4.71-4.69 (m, 2H), 4.54 (d, 1H, J = 11.7 Hz), 4.51-4.43 (m, 1H), 4.42 (d, 1H, J = 6.0 Hz), 4.39-4.28 (m, 2H),

4.14 (dd, 1H, J = 13.5, 7.0 Hz), 4.07-4.03 (m, 1H), 3.91-3.89 (m, 1H), 3.86-3.82 (m, 2H), 3.71 (s, 3H), 3.66- 3.63 (m, 2H), 2.27-2.17 (m, 2H), 2.07-1.91 (m, 2H). 13 C-NMR (50 MHz, CD 3 OD): 174.3, 172.4, 172.2, 139.4, 129.4, 129.1, 128.9,

88.5, 81.8, 80.7, 74.1, 73.9, 73.5, 73.3, 71.7, 60.9, 58.3, 52.8, 46.6, 30.4, 25.8. IRv (cm -1) KBr: 3356, 2880, 1714, 1681, 1651, 1614, 1538, 1455, 1312, 1227, 1198, 1138, 1025, 911, 883, 742, 699. [a] D20 +79.6 (c 0.6, MeOH). Appearance: white solid (mp 47-48 ° C). Yield: 90% after purification by column chromatography (eluent: hexane / ethyl acetate). HRMS calcd for C23H30N2O9 [M + 1]: 478.1951; Found 479.2291.

5th

The above description of the present invention as well as the examples which have been presented are for illustration purposes and do not limit the invention to the form disclosed and exemplified herein. In consequence; variations and modifications consistent with the above teachings; and the skill or knowledge of the relevant technique; are within the scope of the present invention. The above described and exemplified embodiments are intended to further explain known methods for practicing the invention and to enable those skilled in the art to use the invention in such; or others; embodiments and with various modifications required by the specific applications or uses of the present invention. It is intended that the present invention include all modifications and variations thereof; within the scope described in the report and the appended claims.

Claims (9)

1. Polymerase and serine protease inhibitor compounds having the general formula I, where R is defined as amino esters derived from the amino acids Lfenylalanine (L-Phe), L-valine ( L-VaI) 1 L-Glycine (L-GIy) 1 L-Proline (L-Pro) 1 Laryngine (L-Arg) and L-Lysine (L-Lys). Z being benzyl (-Bn), carboxybenzyl (-Cbz) or carboxy-f-butyl (-Boc), hydrogen, alkyl or aryl: Compounds according to claim 1, characterized in that Z is preferably benzyl. Pharmaceutical compositions characterized in that they contain a pharmaceutically acceptable amount in their formulations as active ingredients, the compounds described in claims 1 and 2, as well as their derivatives and / or isolated or combined salts. Pharmaceutical compositions according to claim 3, characterized in that they have an active concentration of inhibitors of serine proteases and DNA and RNA polymerases, ranging from 10μΜ to 400μΜ. Pharmaceutical compositions according to Claim 4, characterized in that they have a preferential concentration of 10μΜ to 400 μΜ. Pharmaceutical compositions according to Claim 3, characterized in that the excipients employed are: adjuvants, sweeteners, solvents, stabilizers, antioxidants, preservatives, buffers, flavorants, surfactants, humectants, lubricants, dispersants. Pharmaceutical compositions according to Claim 3, characterized in that the excipients employed are provided for the production of antiviral drugs in pharmaceutical forms intended for external, oral or parenteral use. Pharmaceutical compositions according to claim 7, characterized in that they are used in the preparation of medicaments for the treatment of diseases caused by flaviviridae viruses, viruses of this family comprising hepatitis C, dengue, yellow fever, West fever or Nile fever. Pharmaceutical compositions according to claim 9, characterized in that they are used more specifically in the treatment of diseases caused by hepatitis C virus (HCV) and dengue virus.