CN103304551B - Hepatitis AG14361 - Google Patents

Abstract

The present invention relates to a kind of 4,4 ' double (2 (base of (S) pyrrolidines 2) base of 1H imidazoles 4) biphenyl compounds and its derivative, compound of the present invention is HCV AG14361s, and the activity with HCV-Ab IgG can be used to treat HCV infection patient.

Description

Hepatitis AG14361

Technical field

The present invention relates to antiviral compound, suppress what is encoded by HCV (HCV) more particularly, to one kind The compound of NS5A polymerases (herein, also referred to as serine protease), and the composition containing the compound, and suppress The method of NS5A polymerases.

Background technology

Hepatitis C is the class through blood born caused by HCV (hepatitis C virus, HCV) Liver diseases, patient has the symptoms such as fatiguability, Nausea and vomiting.HCV infection person there are about 60%-80% and develop into chronic hepatitis, Wherein about 20% develops into cirrhosis, and the patient for having about 2%-5% dies from cirrhosis and liver cancer (Modi caused by HCV infection AA,Liang TJ.Hepatitis C:a clinical review.Oral Dis[J].2008,14(1):10-4.).The whole world In the range of, HCV infection number accounts for the 2.2-3.0% of total number of people, about 1.3-1.7 hundred million (Lavanchy D.The global burden of hepatitis C[J].Liver Int 2009；29Suppl s1:74–81.).The infection rate of China HCV is about It is 3.2%, HCV infection is just turning into problem (Chinese Journal of Hepatology, 2004,12 (4) of harm human health:194—198.).

At present, due to lacking specific effective medicine and vaccine, clinically the therapeutic scheme of standard is to use IFN-α (interferon alfa, INF- α) or glycol interferon (pegylated interferon, peg-INF) and sharp bar Wei Lin (ribavirin, RBV) drug combination treats hepatitis C, even if however, by comprising polyethyleneglycol modified (pegylated) the experimental treatment scheme of the combination of alpha-interferon and Ribavirin, most patients are not reduced persistently Viral load amount.There is the shortcomings for the treatment of cycle long, poor selectivity, erious adverse reaction；Interferon and Ribavirin can cause patient There is influenza-like symptom, tired, cognition dysfunction, depression, fash, gastrointestinal symptom, thyroid dysfunction, retinopathy Become, the adverse reaction such as hemolytic anemia and haemocyte are reduced, some patientss is not resistant to and stopped treatment (Manns in advance MP,Wedemeyer H,Cornberg M.Treating viral hepatitis C:efficacy,side effects, and complications[J].Gut.2006；55(9):1350-9).

Thus, the need for there is the limited method for treating HCV infections of clear and definite and unmet exploitation. The protease inhibitors or AG14361 of HCV duplications can directly be suppressed turns into the new thing of anti-hepatitis c virus Matter.Wherein with HCV NS3/4A serine proteases (NS3/4A serine protease, NS3/4ASP), NA5A polymerases and RNA polymerase (NS5B RNA-dependent RNA polymerase, NS5B RdRp) that NS5B RNA are relied on etc. is target, The specific target spot antiviral therapies of searching HCV (specifically targeted antiviral therapy for HCV, STAT-C) medicine is the important directions of HCV-Ab IgG research.

HCV is a kind of positive chain RNA virus.Based on the similitude of wide hair in the amino acid sequence and 5 ' non-translational regions inferred Compare, said that HCV is categorized as individually being planted in flaviviridae.The member of whole flaviviridaes has encapsulated virion, this A little virion include positive chain RNA genome, and it encodes all known virus by the translation of single continuous open reading frame Specific protein.

In HCV genomes, significant heterogeneity is there is in nucleotides and coded amino acid sequence.Table Six kinds of main genotype are levied, and describes more than 50 kinds of hypotype.In the whole world, the main genotypes of HCV are distributed at it Aspect is different, although and numerous studies possible effect of the genotype to pathogenesis and treatment method, the something lost of HCV Pass heterogeneous clinical meaning still unpredictable.

Unit chain HCV rna gene groups are about 9500 nucleotides in length, and with about 3000 amino acid of coding Single large-scale polyprotein single open reading frame (ORF).In infected cell, this polyprotein is in multiple sites by cell Protease and virus protease cracking are so as to produce structural and unstructuredness (NS) protein.For HCV, ripe non-knot The generation of structure protein (NS2, NS3, NS4A, NS4B, NS5A and NS5B) is influenceed by two kinds of virus proteases.The first Protease is cracked at NS2-NS3 tie points；Second protease is the serine stretch protein being contained in the N- end ranges of NS3 Enzyme, and all then cracking in NS3 downstreams are mediated, it is cis at NS3-NS4A cracking sites, for remaining NS4A- NS4B, NS4B-NS5A, NS5A-NS5B site, are trans.NS4A protein seems to play various functions, used as NS3 protease Confactor and potentially contribute to the film of NS3 and other rdrp virus components and position.The complexing of NS3 protein and NS4A Type is required for the processing of effective polyprotein, enhances the proteolytic cleavage at all sites.NS3 protein Also show nuclear nucleoside triphosphatase and DBPA activity.NS5B is a kind of RNA polymerase for depending on RNA, and it is related to HCV Duplication.

Current STAT-C medicines are the important directions of HCV-Ab IgG research, and such medicine is just actively studied by external many pharmaceuticals Thing, some drugs enter into clinical experimental stage, the NS3/4A serpins of Vertex companies of the U.S. Telaprevir (VX-950) and the NS3/4A serine stretch protein enzyme levels of Schering Plough company (Schering Plough) research and development Agent boceprevir (SCH 503034) was ratified to list in 2011 by FDA.NS5A albumen and NS5B AG14361 medicines Thing carries out clinical experimental study.Additionally, also have some trial drugs with other viral enzymes, such as helicase, NS5A and The internal ribosome entry site (IRES) of HCV 5' noncoding regions shows effective disease-resistant toxic effect as target position, part research Really.PPI-461 is to be used to treat with one kind of Stanford and Rutgers universities R ＆ D Cooperations by Presidio pharmaceutical companies The small molecule NS5A/NS4B inhibitor of HCV infection, enters II clinical trial phases at present.It imitates a kind of inhibition peptides blocking Virus N S5A anchors to HCV nucleoprotein with (6th Ann BioPartner N on the combination of cell membrane America(Vancouver),2008；Company Web Page,Presidio,31Jul&13Nov 2008； Pressreleases,Presidio,7Jul 2009&27Apr 2010；BioWorld Today,13Jul 2009).BMS- 790052 is that the hepatitis C virus NS 5 A for treating HCV infection researched and developed by Bristol-Myers Squibb (BMS) company presses down Preparation.Currently carry out phase II clinical trials (Study of BMS-790052Add-On to Standard of Care in Treatment Naive Subjects(HEPCAT)[EB/OL].(2010-9-9)http:// clinicaltrials.gov/ct2/show/NCT01125189)。

The content of the invention

On the one hand, present invention aim at a kind of compound with logical formula (I) structure of offer

Or its officinal salt, wherein：

R₁And R₃Be each independently selected from hydrogen, alkoxy, alkyl, aryl-alkoxy carbonyl, carboxyl, haloalkyl, halogen, Hydroxyl and-NR^aR^bCarbonyl and trialkylsilylalkoxyalkyl；

R₂Selected from C1-C5 low alkyl groups, Alkylcarbonylalkyl, aryl, aryl alkenyl, alkoxy aryl, aryl alkyl, virtue Base epoxide alkyl, cycloalkyl, cycloalkyloxyalkyl, haloalkyl and heterocyclic radical.

In a preferred embodiment, wherein R₁And R₃It is each independently selected from hydrogen.

In another preferred scheme, wherein R₂Alkyl, cycloalkyl selected from C1-C5, aryl, aryl alkyl, cycloalkyl Alkyl, heterocyclic radical and cycloheteroalkylalkyl.

Which salt salt form all pharmaceutical acceptable salts of the present invention including the compound, officinal salt is, therein Counter ion counterionsl gegenions do not have notable contribution to the physiologically active or toxicity to the compound, thus play pharmacological equivalents. These salt can using commercial reagent according to ordinary post technology prepare, some anionic salt forms include acetate, Acistrate, benzene sulfonate, bromide, chloride, citrate, fumaric acid eye, glucuronate salt, hydrobromate, hydrochloric acid Salt, hydriodate, iodide, lactate, maleate, mesylate, nitrate, embonate, phosphate, butanedioic acid Salt, sulfate, it is exactly hydrochlorate, toluene fulfonate and xinofoate, it is therefore preferable to dihydrochloride.Some cation salt bags Include ammonium, aluminium, N, benzathine, bismuth, calcium, choline, diethylamide, lithium, magnesium, meglumine, 4- phenylcyclohexylamines, The salt of piperazine, potassium, sodium, tromethamine and zinc.

General formula compound can be prepared by means known in the art, including including those described below.Some reagents Box intermediate is known in the art, and other kit intermediates can be using easily available starting materials by known in the art Method prepare.Variable (such as digitized " R " replaces base) for describing general formula compound synthesis is used only for illustrating It is bright how to prepare, without should with specification claim or other parts use confounding of variable, the contracting used in scheme Write the convention for generally being used according to this area.

In the particular embodiment, the present invention relates to the compound of following structural：

Or its officinal salt.

The management that in general abbreviation used in scheme uses according to this area, some examples are as follows：THF means four Hydrogen furans；DMF means N,N-dimethylformamide；RCM means closed loop methasis；Boc means tert-butoxycarbonyl；TFA anticipates Refer to trifluoroacetic acid；DMA means DMAC N,N' dimethyl acetamide；PPh₃Mean triphenylphosphine；OAc means acetic acid esters；Me means methyl； COD (or cod) means 1,5- cyclo-octadiene；DCM means dichloromethane；DIPEA means N, N- diisopropylethylamine；Dtbpy anticipates Refer to the pyridine of 4,4 '-di-t-butyl -2,2 '-two；Dba means dibenzalacetone；EDCI means 1- ethyls -3- (3- dimethylamine third Base) carbon diamine hydrochloride；Aq means the aqueous solution；TBUty or t-Bu represent the tert-butyl group, and SEM represents-(trimethyl silyl) second Epoxide methyl；TFA represents trifluoroacetic acid；Ph means phenyl；OAc means acetate；Cbz means benzyloxycarbonyl group；Bn means benzyl； EtOH means ethanol；MeOH means methyl alcohol；TBTU means 2- (1H- BTA -1- bases) -1,1,3,3- tetramethylurea cations Tetrafluoroborate；DMSO means dimethyl sulfoxide (DMSO)；HATU means O- (7- azepine benzos triazol-1-yl)-N, N, N ', N '-tetramethyl Base uronium hexafluorophosphate；TEA means triethylamine；HOBt means I-hydroxybenzotriazole, CH₃CN means acetonitrile；Pd (dppf) Cl2 means double (diphenylphosphine) ferrocene of 1,1'-] palladium chloride.

It is still another object of the present invention to provide a kind of composition, the composition is the present invention containing therapeutically effective amount The logical formula (I) compound or pharmaceutically acceptable salt thereof, and pharmaceutical carrier, diluent or excipient.Therapeutically effective amount, refers to be enough to The total amount of each active component of patient benefit is shown, viral load is such as reduced.Single active component is applied to when individually giving When, therapeutically effective amount refers to single component, when drug combination, refers to and produces the active component of therapeutic effect and ice-cold, but regardless of being No is being given in combination successively or simultaneously.Terms used herein " pharmaceutically acceptable ", the such compound of essence, raw material, Composition and/or formulation, they are in correct medical judgment category, it is adaptable to patient tissue contacts, and without excessive toxicity, Excitant, allergic reaction or other problems or complication, this is thought of as with rational interests/Hazard ratio, and they are made a reservation for Purposes is effective.

Pharmaceutical preparation can exist with unit dosage forms, and its per unit dose contains the active component of scheduled volume, for pre- In the monotherapy of the disease that anti-and treatment HCV is connected to, about 0.01-200 milligrams per kilogram body weight is daily, preferably 0.05- The daily presently disclosed compound dosage level of 100mg/kg body weight.Typically, the Pharmaceutical composition of the disclosure will be with every Day about 1- about 5 this gives, or is given as the mode of continuous infusion.Such administration can be used as chronic or formulation therapy, can With combined with support material with produce single formulation active component amount by according to the disease, the seriousness of the state of an illness treated, Administration number of times, method of administration, the discharge rate of compound used therefor, the course for the treatment of, and age of patient, sex, body weight and body shape Condition and change.Preferred unit dose formulations are containing daily dose or such as hereinbefore sub-doses, or its suitable divided dose Active component.Treatment can be started with being significantly less than the low dose of the dose,optimum of the compound, and hereafter, dosage is according to small Increment increase, until reach optimum efficiency in this case, in general, optimal the giving of the compound is logical Viral effective result, the concentration level without causing any harmful or poisonous side effect will be often provided.

When the compound of the composition comprising the disclosure of the disclosure and at least one other treatment or the combination of prophylactic When, the treatment of compound and two cancers or prophylactic are ined succession and are generally existed with the dosage level between about 10% to 150%, generally more It is preferred that in monotherapy scheme with about 10% to 80% between dosage give.

May be adapted to give pharmaceutical formulation with any appropriate approach, for example by oral (including buccal or sublingual administration), Rectum, intranasal, part (including buccal, sublingual or transdermal), vagina or it is parenteral (including subcutaneous, intracutaneous, intramuscular, in joint, In synovial membrane chamber, in breastbone, interior intrathecal, focus, intravenous or intracutaneous injection or infusion) approach.Can be by known to pharmaceutical field The non-method of people, for example, be mixed with such preparation by by active component and carrier or excipient.It is preferred that oral or be administered to Give.

Pharmaceutical preparation suitable for orally giving can exist as discrete unit, such as capsule or tablet；Powder or particle Agent；Solution or supensoid agent in aqueous or on-aqueous liquid；Edible foaming agent or whips；Or oil-in-water liquid emulsion or oil Bag aqueous emulsion.For example for oral administration in the form of tablets or capsules, active pharmaceutical ingredient can be with oral, atoxic medicine The acceptable inert carrier such as mixing such as ethanol, glycerine, water on.Powder is broken into suitable fine powder size by by compound powder And mix to prepare with the pharmaceutical carrier (such as edible carbohydrate, starch or mannitol) similarly crushed.Flavor enhancement, anti-corrosion Agent, dispersant and colouring agent also there may be.

And, when wanting or needing, suitable adhesive, lubricant, disintegrant and colouring agent can also be mixed and mixed In thing, suitable adhesive includes starch, gelatin, natural sugar such as glucose or beta lactose, corn sweetener, natural and synthesis Gummy gum arabic, bassora gum or mosanom, carboxymethylcellulose calcium, polyethylene glycol etc..For the lubrication of these types Agent includes enuatrol, sodium chloride etc., disintegrant including but not limited to starch, methylcellulose, agar, xanthans etc., for example By preparing mixture of powders, fritter, addition lubricant and disintegrant and tabletted, configuration tablet are granulated or made.Merits and demerits The compound that crushing will be reverberated mixes with diluent or matrix as described above, and optionally with adhesive such as carboxymethyl cellulose Element, alginates, gelatin or with vinylpyrrolidone, retarder such as paraffin, reabsorb accelerator such as quaternary ammonium salt, or absorbent Such as bentonite, kaolin or Dicalcium Phosphate mix, and prepare mixture of powders.Can by with adhesive such as syrup, gelatinized corn starch, The solution wetted powder mixing of Acadia's mucus or cellulose or polymer raw material, and it is pressed through screen cloth granulation.As Another selection of granulation, by mixture of powders by tablet press machine, and can crush granulating by the block of imperfect formation.Can use and add The method of stiffened resin acid, stearate, talcum powder or vegetable oil is particle lubrication in case adhering of tablets is on tablet-forming dies, Then the mixture that will be lubricated is tabletted.Can provide be made up of the sealant of shellac transparent or opaque protective layer, The coating or wax polishing of sugar or polymer raw are coated.

The liquid oral such as solution of dosage unit form can be prepared as, high degradation and elixir, so, the amount given is carried Compound containing scheduled volume.Be dissolved in the suitable aqueous solution processed by flavoring by by compound, can prepare syrup, And by using avirulent solvent, elixir can be prepared, can also add the different tristearin of solubilizer and emulsifying agent such as ethoxylation Alcohol and polyoxyethylene sorbitol ether, preservative, flavoring addition such as peppermint oil or natural sweetener or saccharin or other artificial Flavouring etc..

In one embodiment, said composition can also further include at least one compound with anti-HCV activity, the work Property compound can be interferon, Ribavirin, HCV metal protease inhibitors, HCV serpins, HCV gather Synthase inhibitor, HCV helicase inhibitors, HCV NS4B protease inhibitors or HCV NS5A protease inhibitors etc..

Such as, described interferon, can be selected from interferon alpha 2 b, Interferon a2a, interferon lambda, interferon beta, interference Plain γ, Peg-IFN alpha-2b α, long-acting interferon, Interferon Alfacon-1, lymph yeast cells sample interferon-tau etc..

The method of patient's HCV infection is treated it is still a further object of the present invention is to provide a kind of, including patient is given is treated The compound or its officinal salt of the logical formula (I) of effective dose.

In the method for the treatment of HCV infection patient, still further comprise and give compound described in logical formula (I) or it can medicine Before with salt, afterwards or it is administered simultaneously at least one other compound with anti-HCV activity.

Table 1 below lists the example of some exemplary compounds that can be given together with disclosure compound, is controlled combining In treatment, the present invention disclose compound can with other anti-HCV reactive compounds or and together with or separate or by general Each compound mixing gives together in the composition.

Table 1

Brief description of the drawings

Fig. 1 is compound K W811's¹H-NMR collection of illustrative plates.

Fig. 2 is the LC-MS collection of illustrative plates of compound K W811.

The antiviral activity figure of Fig. 3 compound Ks W811.

Fig. 4 is the cytotoxicity figure of compound K W811 and KW819.

Embodiment

The real scheme is not limitation scope disclosed by the invention, conversely, the disclosure covers and all of such as may include Alternative, amendment and equivalent in Claims scope.Thus, the embodiment including specific embodiment once will Illustrate a kind of practice of the disclosure, it should be appreciated that these embodiments are, in order to illustrate the purpose of some embodiments, and to present Out believe to the description of its method and concept it is most useful and understandable with offer.

The synthesis of embodiment 1 (S) -2- (methoxyl group carbon acylamino) -2- phenylacetic acids

By ClCO₂Me (41.4mmol) is added drop-wise to (the S) -2- aminophenyl acetic acids tert-butyl ester/HCl of cooling (ice/water) In half solution of (40.52mmol) and diisopropylethylamine (81.52mmol) and THF (410ml), and stir at the same temperature Mix 5.5 hours, after volatile ingredient is removed in vacuum, residue is distributed between water (100ml) and ethyl acetate (200ml), There are several layers of use 1N HCl (25ml) and saturation NaHCO₃(30ml) is washed, anhydrous Na₂SO₄Dry, be concentrated in vacuo after filtering, gained Colorless oil hexanes trituration, is washed after filtering with hexane (100ml), obtains (R) -2- (methoxycarbonylamin) -2- phenyl Tert-butyl acetate (7.7g, white solid).

In CH2Cl2 (160ml) solution for the cooling (ice/water) that TFA (16ml) is added drop-wise to above-mentioned product, cooling is removed After bath, stirring reaction mixture 20 hours, because deprotection reaction is still to complete, therefore in addition TFA (1.0ml), and followed by Continuous stirring 2 hours, after volatile ingredient is removed in vacuum, gained oiliness residue with diethyl ether (15ml) and hexane (12ml) are processed, Be precipitated thing, sediment after filtering, with ether/hexane (about 1:3,30ml) after washing, vacuum drying obtains (S) -2- (methoxyl group carbon acylamino) -2- phenylacetic acids.Optical activity:- 176.9 ° [in c=3.7mg/ml, H2O；λ=589nm].HNMR (DMSO-d6, δ=2.5ppm, 400MHz)：δ 12.84 (br s, 1H), 7.96 (d, J=8.3,1H), 7.41-7.29 (m, 5H), 5.14 (d, J=8.3,1H), 3.55 (s, 3H).LC/MS:For [M+H]⁺C₁₀H₁₂NO₄Measured value is 210.1.

Embodiment 2

Methyl (S) -2- ((S) -2- (4- (4 '-(2- ((S) -1- (3- methy oxetane -3- carbonyls) pyrrolidines -2- Base) -1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidin-1-yl) -2- oxo -1- phenylethylcarbamate first Acid benzyl ester.

Synthesis step

Step a：The bromo- 1- of 2- (4- bromobenzenes)-ethyl ketone

1- (4- bromobenzenes)-ethyl ketone (50g, 0.253mmol) is dissolved in dichloromethane (300mL), bromine liquid is added (40.81g, 0.225mol), is filling N₂Under the conditions of, mixture is stirred at room temperature 4 hours, then concentrates reactant, obtains The bromo- 1- of product 2- (4- bromobenzenes)-ethyl ketone, gained crude product (50g, 70%) synthesizes for next step, and crude product is without further Purifying.

Step b：(S) -1- benzyls -2- (2- (4- bromobenzenes) -2- oxoethyls) pyrrolidines -1,2- dicarboxylic acid esters

By the bromo- 1- of 2- (4- bromobenzenes)-ethyl ketone (9.5g, 36mmol) and (S) -1- (benzyloxycarbonyl group) pyrrolidines -2- formic acid In DIPEA (4.6g, 38mmol) is added at 20 DEG C, mixed liquor is stirred overnight the acetonitrile solution of (9g, 38mmol) mixture, Gained reactant is washed with strong brine, organic phase anhydrous sodium sulfate drying, and is concentrated to give product (15.3g, 93.6%).Institute Obtain product to synthesize for next step, without being further purified.

Step c：(S)-benzyl -2- (4- (4- bromobenzenes) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters

In (S) -1- benzyls -2- (2- (4- bromobenzenes) -2- oxoethyls) pyrrolidines -1,2- dicarboxylic acid esters (15g, In toluene solution 33.7mmol), ammonium acetate, mixture is added to be heated to 100 DEG C and kept for 5 hours, the concentration of gained mixture is residual Thing silica gel chromatography is stayed, product (S)-benzyl -2- (4- (4- bromobenzenes) -1H- imidazoles -2- bases) pyrrolidines -1- first is obtained Acid esters (5g, 35%).

Step d：(R) -2- (4 ' -2- ((R) -1- ((R) -2- (methoxyl group carbon acylamino) -3- methylbutyryls base) pyrrolidines - 2- phosphinylidynes oxygen) biphenyl -4- bases) -2 oxoethyls

The 1 of (S)-benzyl -2- (4- (4- bromobenzenes) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters (6g, 14mmol), Potassium acetate (4.15g, 42mmol) and connection boric acid pinacol ester (4.15g, 42mmol) are added in 4- bis- oxazole (60ml) solution.It is mixed Compound deaerates and uses N₂Displacement 3 times, is subsequently adding Pb (dppf) Cl₂.Mixture stirs 3h, gained reactant mixture at 80 DEG C Wash with water and extracted with EA, mix organic phase anhydrous Na₂SO₄Dry, concentrate, residue is obtained after silica gel chromatography To (R) -2-, (4 ' -2- ((R) -1- ((R) -2- (methoxyl group carbon acylamino) -3- methylbutyryls base) pyrrolidines -2- phosphinylidynes oxygen) join Benzene -4- bases) -2 oxoethyls (6g, 90%).

Step e：(S) -2- (2- (4- bromobenzenes) -2- oxoethyls) 1- tert-butyl pyrrolidine -1,2- dicarboxylic acid esters

Products therefrom (9.5g, 36mmol) and (S) -1- (tertbutyloxycarbonyl) pyrrolidines -2- carboxylic acids in step d (9g, In acetonitrile solution 38mmol), in adding DIPEA (4.6g, 38mmol), gained mixture to be stirred overnight at 20 DEG C, reaction is mixed Compound is washed with strong brine, and crude product (S) -2- (2- (4- bromobenzenes) -2- are obtained after organic phase anhydrous Na 2SO4 dryings, concentration Oxoethyl) 1- tert-butyl pyrrolidine -1,2- dicarboxylic acid esters (15.3g, 93.6%).Products obtained therefrom synthesizes for next step, nothing Need to be further purified.

Step f：(S)-tert-butyl group 2- (4- (4- bromobenzenes) -1H- imidazoles -2 base) pyrrolidines -1- formic acid esters

In step e in the toluene solution of products obtained therefrom (15g, 33.7mmol), ammonium acetate (15g, 33.7mmol) is added.It is mixed Compound is heated to 100 DEG C and keeps 5h, the concentration of gained reactant mixture, and product is obtained after residue silica gel chromatography (S)-tert-butyl group 2- (4- (4- bromobenzenes) -1H- imidazoles -2 base) pyrrolidines -1- formic acid esters (5.6g, 35%).

Step g：(S)-benzyl 2- (4- (4 '-(2- ((S) -1- (tertbutyloxycarbonyl) pyrrolidin-2-yl) -1H- imidazoles -4- Base) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters

Products obtained therefrom (2.08g, 5.285mmol) in step f, and (R) -2- (4 '-(2- ((R) -1 ((R)-(methoxy carbon Acylamino-) -3- methyl determines acyl group) pyrrolidines -2- phosphinylidynes epoxide) acetyl group) biphenyl -4- bases) -2- oxoethyls (3g, In DME solution 6.34mmol), Na is added₂CO₃Solution (2M, 5ml).Resulting solution deaerates and uses N₂Displacement three times, adds Pb (PPH3) 2Cl2, gained reaction solution is stirred 16 hours at 87 DEG C, and reaction solution is cooled to environment temperature and is diluted with ethyl acetate, institute Obtain organic phase anhydrous Na₂SO₄Dry, concentrate, residue obtains product (S)-benzyl 2- (4- after silica gel chromatography (4 '-(2- ((S) -1- (tertbutyloxycarbonyl) pyrrolidin-2-yl) -1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) Pyrrolidines -1- formic acid esters (2.5g, 72%).

Step h：(S)-tert-butyl group 2- (4- (4 '-(2 ((S)-pyrrolidin-2-yl) -1H- imidazol-4 yls) -1H- imidazoles -2- Base) pyrrolidines -1- formic acid esters.

In step f in the methanol solution of products therefrom (2.5g, 3.8mmol), 10% Pd/C (1.5g) is added.Mixture Degassing, and use H₂After displacement three times, reaction solution is stirred at room temperature 2 days, then reactant mixture filtering, after filtrate concentration To product (S)-tert-butyl group 2- (4- (4 '-(2 ((S)-pyrrolidin-2-yl) -1H- imidazol-4 yls) -1H- imidazoles -2- bases) pyrroles Alkane -1- formic acid esters (1.5g, 75%).

Step i：(S)-tert-butyl group 2- (4- (4 '-(2- ((S) -1- ((S) -2- (methoxy carbon acylamino) -2- phenylacetyls) pyrroles Cough up alkane -2- bases) 1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters

Products obtained therefrom (1g, 1.9mmol) and EDCI (0.4374g, 2.28mmol) and HOBt in step h (0.3092g, In acetonitrile solution 2.28mmol), (S) -2- (methoxy carbon acylamino) -2 phenylacetic acids (476mg, 2.28mmol) are added.Mixed liquor 0 DEG C is cooled to, and DIPEA (490mg, 3.8mmol) is added dropwise in the case where being kept for 10 DEG C.Gained reaction solution was stirred at room temperature Night, reactant mixture is washed with water, and uses CH₂Cl₂Extraction, organic phase anhydrous Na₂SO₄Dry, concentration, residue is through silica gel Column chromatography is purified, and obtains product (S)-tert-butyl group 2- (4- (4 '-(2- ((S) -1- ((S) -2- (methoxy carbon acylamino) -2- Phenylacetyl) pyrrolidin-2-yl) 1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters (0.8g, 60%), it is yellow solid.

Step j：Methyl (S) -2- oxo -1- phenyl -2- ((S) -2- (4- (4 '-(2- ((S)-pyrrolidin-2-yl) -1H- The base of imidazoles -4) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidin-1-yl) ethyl ester.

In step i in the DCM solution (5mL) of products obtained therefrom (0.5g, 0.7mmol), TFA (3mL) is added dropwise over, gained is mixed Compound is stirred overnight at room temperature, and after being detected through TLC, reaction gained mixture is washed with NaHCO3, and is extracted with DCM, organic relevant It is dry, it is concentrated to give yellow solid methyl (S) -2- oxo -1- phenyl -2- ((S) -2- (4- (4 '-(2- ((S)-pyrrolidines -2- Base) base of -1H- imidazoles -4) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidin-1-yl) ethyl ester (0.4g, 93%).

Step k：Methyl (S) -2- ((S) -2- (4- (4 '-(2- ((S) -1- (3- methy oxetane -3- carbonyls) pyrroles Alkane -2- bases) -1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidin-1-yl) -2- oxo -1- phenylethyls Benzyq carbamate.

Products therefrom (0.35g, 1.9mmol) in step j, EDCI (0.4374g, 0.8mmol), HOBt (0.3092g, In acetonitrile liquid (5mL) 0.8mmol), 3- methy oxetane -3- formic acid esters (0.093g, 0.8mmol) is added.Gained is mixed After compound is cooled to 0 DEG C, DIPEA (0.172g, 1.336mmol) is added dropwise under the conditions of keeping temperature is less than 10 DEG C.Gained Mixture is stirred overnight at room temperature, and gained reactant is washed with water, and uses CH₂Cl₂Extraction, organic phase anhydrous Na₂SO₄Dry, it is dense Contracting, residue is purified through p-TCL, obtains yellow solid product：Methyl (S) -2- ((S) -2- (4- (4 '-(2- ((S) -1- (3- first Base oxetanes -3- carbonyls) pyrrolidin-2-yl) -1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines - 1- yls) -2- oxo -1- phenylethylcarbamates benzyl formate (100mg, 15%), its HNMR collection of illustrative plates and LC-MS collection of illustrative plates see Fig. 1 and 2。

Embodiment 3

The synthesis of compound K W810

(S)-tert-butyl group 2- (4- (4 '-(2- ((S) -1- ((S) -2- (methoxy carbon acylamino) -3- methylbutyryls base) pyrroles Alkane -2- bases) 1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters synthesis

By products obtained therefrom (1g, 1.9mmol) and EDCI (0.4374g, 2.28mmol) and HOBt in the step h of embodiment 2 In the acetonitrile solution of (0.3092g, 2.28mmol), (S) -2- (methoxy carbon acylamino) -3 methylbutanoic acids (2.28mmol) are added. Mixed liquor is cooled to 0 DEG C, and is added dropwise over DIPEA (490mg, 3.8mmol) in the case where being kept for 10 DEG C.Gained reaction solution is at room temperature It is stirred overnight, reactant mixture is washed with water, and uses CH₂Cl₂Extraction, organic phase anhydrous Na₂SO₄Dry, concentration, residue Through silica gel chromatography, product (S)-tert-butyl group 2- (4- (4 '-(2- ((S) -1- ((S) -2- (methoxy phosphinylidyne ammonia is obtained Base) -3- methylbutyryls base) pyrrolidin-2-yl) 1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- first Acid esters (0.8g, 60%), is yellow solid.

Remaining step obtains product KW810 with step j and k in embodiment 2：Methyl (S) -2- ((S) -2- (4- (4 '-(2- ((S) -1- (3- methy oxetane -3- carbonyls) pyrrolidin-2-yl) -1H- imidazol-4 yls) biphenyl -4- bases) - 1H- imidazoles -2- bases) pyrrolidin-1-yl) (0.05g, purity is more than -2- oxo -3- methylbutanoylaminos methyl formate 95%) it is 679.81 that, ES-API collection of illustrative plates determines its value.

Embodiment 4

The synthesis of compound K W812

(S)-tert-butyl group 2- (4- (4 '-(2- ((S) -1- ((S) -2- (methoxy carbon acylamino) -4- methylvaleryls) pyrroles Alkane -2- bases) 1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- formic acid esters synthesis

By products obtained therefrom (1g, 1.9mmol) and EDCI (0.4374g, 2.28mmol) and HOBt in the step h of embodiment 2 In the acetonitrile solution of (0.3092g, 2.28mmol), (S) -2- (methoxy carbon acylamino) -4- methylvaleric acids (2.28mmol) are added. Mixed liquor is cooled to 0 DEG C, and is added dropwise over DIPEA (490mg, 3.8mmol) in the case where being kept for 10 DEG C.Gained reaction solution is at room temperature It is stirred overnight, reactant mixture is washed with water, and uses CH₂Cl₂Extraction, organic phase anhydrous Na₂SO₄Dry, concentration, residue Through silica gel chromatography, product (S)-tert-butyl group 2- (4- (4 '-(2- ((S) -1- ((S) -2- (methoxy phosphinylidyne ammonia is obtained Base) -4- methylvaleryls) pyrrolidin-2-yl) 1H- imidazol-4 yls) biphenyl -4- bases) -1H- imidazoles -2- bases) pyrrolidines -1- first Acid esters (0.8g, 60%), is yellow solid.

Remaining step obtains product KW812 with step j and k in embodiment 2：Methyl (S) -2- ((S) -2- (4- (4 '-(2- ((S) -1- (3- methy oxetane -3- carbonyls) pyrrolidin-2-yl) -1H- imidazol-4 yls) biphenyl -4- bases) - 1H- imidazoles -2- bases) pyrrolidin-1-yl) (0.05g, purity is more than -2- oxo -4- methylpentanoylaminos methyl formate 95%) it is 693.84 that, ES-API collection of illustrative plates determines its value.

Embodiment 5

The synthesis of compound K W813

It is raw material with (S) -2- (methoxy carbon acylamino)-butyric acid, its synthesis step obtains product with embodiment 4 It is 665.78 that KW813, ES-API collection of illustrative plates determine its value.

Embodiment 6

The synthesis of compound K W814

It is raw material with (S) -2- (methoxy carbon acylamino)-propionic acid, its synthesis step obtains product with embodiment 4 It is 651.76 that KW813, ES-API collection of illustrative plates determine its value.

The detection of the Compound ira vitro anti-HCV activity of embodiment 6

According to the method (J.Virol.2003 of the reports such as Blight；77:3181-3190) build respectively and contain HCV genes The HCV full-length genome replication in vitro subsystems and real time fluorescent quantitative reverse transcriptase polymerase of hypotype 1a (H77) and 1b (Con1) Chain reaction (qRT-PCR) detection method.The compound of various concentrations and identical is separately added into above-mentioned viral replication system The dimethyl sulfoxide (DMSO) (DMSO, negative control) of volume, after continuing to cultivate 48 hours, carries out qRT-PCR viral nucleic acid quantitative determinations.

Each compound (KW810-KW814) is dissolved in DMSO, and the storing solution for being configured to 10mM is standby, carry out it is antiviral In experiment, the initial concentration of each compound is set in 100nM, final concentration of 0.01pM.10 times are carried out respectively to above-claimed cpd It is serially diluted, each treatment carries out 3 independent repetitions and tests respectively, repeats to set 5 multiple holes every time, and experimental data carries out statistical Analysis.

Under equal conditions, scheme disclosed in the clinical medicine (BMS790052) according to BMS (hundred Mei Shiguibao companies) is entered Row synthesis compound BMS790052, the positive control with the compound as external activity.

After testing, compound is sufficiently close to for the inhibition of the HCV duplicating efficiencies of two kinds of hypotypes of H77 and Con1, with As a example by Con1 hypotypes, the IC that compound K W-811 is replicated to HCV₅₀About 0.1pM (result is shown in Fig. 3), activity is most strong, other chemical combination The IC of the anti-HCV activity of thing₅₀Value the results are shown in Table 2.

The compound antiviral activity of table 2

The Compound ira vitro cytotoxicity experiment of embodiment 7

To obtain the vitro cytotoxicity data of compound, the CytoTox- that we are produced using Promega companies of the U.S. Glo Cytotoxicity Assay kits, according to the method that its specification is recorded, HepG2, uterine neck to hepatocyte origin The cell lines such as the 293T in cancer cells HeLa, hemopoietic origin K562, the A549 in lung source and nephrocyte source are to two The cytotoxicity for opening compound has carried out analyzed in vitro, is as a result displayed under the treatment conditions that highest concentration for the treatment of is 10 μM, not See any cytotoxic effect (result is shown in Fig. 4).

Claims (7)

1. there is the compound of following structure：

Or its officinal salt.

2. the composition of compound or its officinal salt and pharmaceutical acceptable carrier described in claim 1 is included.

3. the composition described in claim 2, further includes at least one other compound with anti-HCV activity.

4. the composition described in claim 3, wherein the described compound with anti-HCV activity be interferon, Ribavirin, HCV metal protease inhibitors, HCV serpins, HCV AG14361s or HCV helicase inhibitors.

5. the composition described in claim 4, wherein the described compound with anti-HCV activity is selected from HCV NS4B protease Inhibitor, HCV NS3 serpins, HCV NS5B AG14361s or HCV NS5A protease inhibitors.

6. utilization of the compound or its officinal salt described in claim 1 in hepatitis medicine is prepared.

7. utilization of the composition described in any one claim in hepatitis medicine is prepared in claim 2-5.