CN106366146A

CN106366146A - 2-dialkyl substituted nucleoside analogue and uses thereof

Info

Publication number: CN106366146A
Application number: CN201510410358.6A
Authority: CN
Inventors: 黄新全; 赵蕾; 刘江; 杨学聪
Original assignee: Zhongke Yunhe (beijing) Bio Pharmaceutical Technology Co Ltd
Current assignee: Zhongke Yunhe (beijing) Bio Pharmaceutical Technology Co Ltd
Priority date: 2015-07-13
Filing date: 2015-07-13
Publication date: 2017-02-01

Abstract

The present invention discloses a 2-dialkyl substituted nucleoside analogue and uses thereof, wherein the 2-dialkyl substituted nucleoside analogue is the nucleoside represented by a general formula I, a nucleoside aminophosphate compound, or a pharmaceutically acceptable prodrug, a salt, a solvate, a stereoisomer, a tautomer or a polymorphic form thereof, and a metabolite thereof. According to the present invention, the preparation is convenient, and the performance is excellent.

Description

Nucleoside analog and purposes that 2 '-bis- alkyl replace

Technical field

The present invention relates to the double alkyl nucleosides analog of 2'- and its prodrug, and it is used for treating hepatitis C viruss (hcv) therapeutic use infecting.The invention still further relates to being used for preparing nucleoside analog disclosed herein and wherein The method of mesosome.

Background technology

Hepatitis C viruss (hcv) are normal chain rna virus.According to estimates, there are about 200,000,000 people infection hcv in the world. The diverse clinical manifestations of infection with hepatitis C virus, gently to inflammation, weight is to liver cirrhosis, hepatocarcinoma.Chronic hepatitis liver Inflammation can also concurrently show outside some livers, including rheumatoid arthritiss, drying property conjunctivokeratitis, flat tongue Moss, glomerulonephritiies, mixed type cryoglobulinemia, b cell lymphoma and porphyria cutanea tarda etc., Caused by being probably the reaction of body abnormal immune.And during hepatitis C cirrhosis patients in decompensation, various complication can occur As ascites abdominal cavity infection, upper gastrointestinal hemorrhage, hepatic encephalopathy, hepatorenal syndrome, liver failure etc. shows. Hcv belongs to flaviviridae hepatovirus virus, and it is pestivirus and Huang with two other genus in flaviviridae The similar drug development having identified many anti-hcv agent for directly working of the gene structure of Tobamovirus Potential molecular target, including ns2-ns3 autologous protein enzyme, n3 protease, n3 unwindase and ns5b gather Synthase.Ns5b rna dependency rna polymerase (rdrp) is that duplication single stranded positive-sense rna genome institute is required , and this enzyme has caused huge interest in the middle of Pharmaceutical Chemist.Core for hcv tool activity Glycosides is ns5b rdrp inhibitor.Nucleoside must be converted to its corresponding triguaiacyl phosphate, and it is incorporated at 3'- end In virus rna, virus rna is made to extend stopping showing chain terminating agent form.Some nucleoside activity are weaker, because For they can not effectively by tyrosine phosphorylation or be not kinases substrate, and some inertia nucleoside when with When chemical mode is converted into its triguaiacyl phosphate, the activity becoming to be directed in vitro some viruses is very strong.Nucleoside phosphorylase Ester (nucleotide) itself very frequently can not be used as medicine, because their envelope cores before entering in cell Thuja acid and other hydrolytic enzyme dephosphorylations or its polarity can not enter in cell by force very much.In order to improve nucleoside Its phosphate prodrugs is studied by biological activity, because they can potentially get around the first of speed limit The phosphorylation of step.Recently, phosphoramidate prodrugs approach has confirmed is that one kind makes the biologically inert Nucleoside be Active single-nucleotide phosphate, thus get around effective ways (the pharmaceutical chemistry magazine of the phosphorylation of the speed limit first step (j.med.chem.,2007,50(22),5463-5470).It has been reported that nucleoside phosphoramidate effectively will Nucleoside 5'- phosplate is delivered to (wo2008/121634 in liver；Wo2008/082601 and wo2008/082602).In recent years, there are many patent applications to disclose and utilized phosphoramidate As nucleoside prodrugs, single-nucleotide phosphate is delivered in tissue, is especially delivered in liver (us6455513、wo2009/052050、wo2008/121634、wo2008/0833101、wo2008/062206、 wo2007/002931、wo2008/085508、wo2007/095269、wo2006/012078、 wo2006/100439).Single-nucleotide phosphate can turn to bisphosphate by phosphoric acid further, and then becomes Biological activity triguaiacyl phosphate.Therefore, the present invention have studied the phosphate prodrugs of synthesized nucleoside simultaneously.

Lucky Leadd B.V hepatitis C medicine Suo Feibuwei (trade name: sovaldi, common name: Sofosbuvir) on December 6th, 2013 obtains food and medicine Surveillance Authority of the U.S. (fda) and ratifies to be used for Gene 1 type, 2 types, the treatment of 3 types and 4 type chronic hepatitis C (hepatitis c) adult patients.Rope Fei Buwei is the first granted medicine that can be used for the full oral treatment regimes of hepatitis C, for specific gene type (2 Type, 3 types) chronic hepatitis C treatment when, the demand to conventional injection interfering effects of drug element (ifn) can be eliminated. Merck drugmaker is being developed for treating the idx21437 of hepatitis C, is also a kind of uridylate Prodrug ns5b inhibitor, its chemical constitution does not also disclose so far.Achillion pharmacy is public Department discloses uridylate prodrug ns5b inhibitor ach-3422.Recently, medivir and idenix In succession individually disclose the double halogen nucleoside analog wo2015081297 in 2 '-position, wo2015034420 and wo2015056213.During these are open, 2 '-position at least substituent group is the strong electron-withdrawing group such as halogen or hydroxyl Group, 2010, wo2010066699 disclosed the nucleoside analog that one group special 2 '-bis- alkyl replace, its In the uridylate precursor analog that replaces of 2 '-cyclopropyl, compound 3, there is certain anti-hepatitis C disease Cytotoxic activity (ec₅₀For 9.6um).Disclosed herein 2 '-bis- alkyl, especially double methyl substituted ucleosides Compound has anti-hepatitis C virus activity well.

Content of the invention

It is an object of the invention to provide a kind of for treat and/or prevent hcv infection formula i shown in New 2 '-bis- alkyl nucleosides analog and its stereoisomer, salt, hydrate, solvate or crystallization and Its purposes.

The technical solution of the present invention is:

A kind of new 2 '-bis- alkyl nucleosides analog synthetic methods and its stereoisomer, salt, hydrate, molten Agent compound or crystallization and application thereof.On the one hand, the present invention is provided to treating 2 '-bis- alkane of mankind hcv infection Yl nucleosides and its phosphate prodrugs with and combinations thereof.In yet another aspect, the present invention is provided to preparation 2 '- The method of double alkyl nucleosides, phosphate prodrugs and its intermediate. The nucleoside analog that 2 '-bis- alkyl replace, is characterized in that nucleoside represented by formula i, nucleoside phosphoramidate Compound or its pharmaceutically acceptable prodrug, salt, solvate, stereoisomer, tautomer or many Crystalline forms, metabolite:

Wherein,

(1) base is natural or engineered miazines or purines nucleoside base；

(2) pd is hydrogen；Or phosphoramidate, phosplate, bisphosphate, triguaiacyl phosphate or it is stable Phosphate prodrugs:

(3)r_a、r_bEach independent, c can be respectively selected from_1-6Alkyl；Replace alkyl；The thiazolinyl of c2-c6, alkynyl Or replace thiazolinyl, alkynyl；Especially methyl or deuterated methyl (cd₃、ch₂d、chd₂)；Or containing extremely The halogenated methyl (- cf of a few halogen₃、-ch₂f、-chf₂、-ccl₃、-ch₂cl、-chcl₂、-cf₂cl、 -ccl₂f、-chfcl)；Or contain an oxygen, nitrogen, phosphorus, the substituent methyl (- ch of sulfur heteroatom₂oh、-ch₂or₁, -ch₂sh、-ch₂sr₁,-ch₂n₃,-ch₂nh₂,-ch₂nhr₁)；

Rc select hydrogen (h), halogen (f, cl, br, i), cyano group (cn), nitrine (n₃)、c_1-6Alkyl, replacement Alkyl, the thiazolinyl of c2-c6, alkynyl or substituted thiazolinyl, alkynyl；

(4) w is oxygen or sulfur；

(5) x and y is each independent, and one of is-h ,-or₁、-sr₁、-nr₁r₂, with n connect natural or Non-natural amino acid estersOr the natural or non-natural amino acid esters being connected with oxygen；And another is then -or₁,-nr₁r₂；

(6) z is oh；Or z is combined with y and constitutes a hexa-member heterocycle, the common table of wherein z and y Show-o- key；And another x is then-or₁、-sr₁、-nr₁r₂, with n connect natural or alpha-non-natural amino acid EsterOr the natural or non-natural amino acid esters being connected with oxygen；

(7) r is natural or the side chain of non-natural alpha-amino acid；r₁For hydrogen, alkyl, cycloalkyl, aryl, miscellaneous Cyclophane base, aryl alkyl, heterocyclic aryl alkyl, alkyl-carbonyl sulfanyl, alkoxycarbonyl alkyl, aralkoxy Carbonylic alkyl, acyl group alkoxyl (aralkyl), naphthene base carbonyl alkoxyl, alkoxycarbonylamino carbonyl Base sulfanyl, hydroxyalkylcarbonyl sulfanyl, acyl group alkoxyl or aminoalkylcarbonyl alkoxy carbonyl group sulfane Base；r₂For hydrogen or alkyl.

Phosphate prodrugs of nucleoside compound that according to formula i 2 '-bis- alkyl replace or its pharmaceutically may be used

Accept salt, hydrate and solvate, preferably from following compound ii to viii:

Wherein,

(1) base is selected from

Or its tautomer

Wherein r₃For h, oh, nh₂,sme,or’,f,cl,br,i,-n₃,nhoh,-nhor’,-nhr”；

r₄For h, oh, or ', sr ', nhr ", f, cl, br, i ,-n₃,nhoh,-nhor’；

r₅For h, oh, nh₂,or’,nhr”,f,cl,br,i,nhoh,-nhor’；

r₆For h, d, oh, or ', nhr ", f, cl, br, i, cn, n₃, nhoh ,-nhor ', c_1-10Non-substituted Alkyl, c_1-10Replace alkyl；

r₇For h, d, oh, or ', sr ', nhr ", f, cl, br, i, cn, n₃；

(2) r ' is selected from c_1-10Substituted or non-substituted alkyl or c_1-10Cycloalkyl；R " is selected from h, c_1-10Replace or non- Replace alkyl, aryl, heterocyclic aryl, Heterocyclylalkyl, cycloalkyl；Ar is substituted or non-substituted phenyl, takes Generation or non-substituted naphthyl or substituted or non-substituted heterocyclic aryl；

(3)w、x、y、z、r、r₁And r₂As defined in claim 1；

(4) phosphorus atoms (p) are chiral centre, can be sp or rp of single configuration, or sp and rp is any The mixture of ratio.

The phosphate prodrugs of nucleoside compound or its medicine that according to formula ii-viii 2 '-bis- alkyl replace Acceptable salt and solvate on, preferably from following structural formula ix to xiii:

Wherein,

Hal is halogen, including fluorine, chlorine, bromine, iodine；Other substituent groups w, x, y, z, base press formula i Defined in.

The phosphate prodrugs of nucleoside compound or its medicine that according to formula i-xiii 2 '-bis- alkyl replace Acceptable salt, hydrate and solvate on, in the molecule shown in its general structure i to xiii arbitrarily Hydrogen atom (h) on position can be replaced by deuterium (d) and remain in that its intrinsic antiviral activity.

The nucleoside compound that according to formula i-xiii 2 '-bis- alkyl replace, the chemical combination below prioritizing selection Thing parent 1001-1084, but it is not limited solely to following compound:

Phosphate prodrugs of nucleoside compound that according to formula i-xiii 2 '-bis- alkyl replace and its different Structure body, the compound 101-133 below prioritizing selection, 201-232,301-332,401-429 but not only It is confined to following compound:

The triguaiacyl phosphate of the nucleoside compound that 2 ' according to formula i-xiii-bis- alkyl replace, below prioritizing selection Compound 501-640, but be not limited solely to following compound:

Phosphate prodrugs of nucleoside compound that according to formula i-xiii 2 '-bis- alkyl replace or it is different Structure body, officinal salt, hydrate, solvate or crystallization and pharmaceutically acceptable carrier, diluent or tax Shape agent is prepared by mixing into pharmaceutical preparation and nanometer formulation, to be suitable for oral or parenteral.Medication Include, but are not limited to Intradermal, intramuscular, intraperitoneal, intravenouss, subcutaneous, intranasal and peroral route.Described system Agent can be applied by any approach, for example pass through infusion or inject, by transepithelial or mucocutaneous (for example Oral mucosa or rectum etc.) approach that absorbs applies.Administration can be whole body or local.Oral administration system The example of agent includes solid or liquid dosage form, specifically, including tablet, pill, granule, powder, capsule Agent, syrup, Emulsion, suspensoid etc..Described preparation can be prepared by methods known in the art, and comprises medicine The conventional use of carrier of thing formulation art, diluent or excipient, nanometer formulation.

A kind of medical composition, the nucleosidation that its 2 '-bis- alkyl in comprising according to formula i-xiii replaces The phosphate prodrugs of compound or its isomer, officinal salt, hydrate, solvate, crystallization and medically may be used The excipient accepting.According to the medical composition of this claim, wherein said other therapeutic agent independence Group and medicine selected from consisting of: ribavirin (ribavirin), interferon, the suppression of hcvns3 protease Preparation, hcv reverse transcriptase ns5b non-nucleosidic inhibitors, hcv reverse transcriptase ns5b nucleosidic inhibitors, ns5a suppression The synergist of preparation and ns5a inhibitor, entry inhibitor, ciclosporin immunosuppressant, ns4a antagonist, Ns4b inhibitor, cyclophilin inhibitor (cyclophilin inhibitor).

According to formula i-xiii, it is used for antiviral for antiviral, including hcv, hbv, hiv, Rsv, norovirus, ev71, dengue fever, influenza virus, the treatment use of Ebola virus, particularly control Treat the application in the medicine of flaviviridae, it is characterized in that: described flaviviridae is hepatitis C viruss.

The method preparing nucleoside analog disclosed herein and its intermediate

Term defines:

Unless otherwise directed, otherwise when referring to compound provided herein, following term has following meanings.

Term " acyl group " or the group of " ester that o- connects " inclusion following formula: alkyl-co or aryl-co or ring-type Alkyl-co.

" alkyl " includes being usually c as the term is employed herein₁To c₂₀Saturated straight chain, side chain or cyclic hydrocarbon, and And specifically include methyl, cf₃、ccl₃、cfcl₂、cf₂Cl, ethyl, ch₂cf₃、cf₂cf₃, propyl group, Isopropyl, cyclopropyl etc..The non-limiting examples that the part of described alkyl can be replaced are selected from and consist of Group: halogen (fluorine, chlorine, bromine or iodine), hydroxyl, amino, alkyl amino, arylamino, alkoxyl, Aryloxy group, nitro, cyano group etc..

" thiazolinyl " includes univalent alkene unsaturated alkyl, in a certain embodiment, has at most 11 carbon atoms, its Can be straight or branched and there is at least one or 1 to 2 alkene unsaturation site.Exemplary thiazolinyl bag Include vinyl (-- ch=ch₂), positive acrylic (-- ch₂Ch=ch₂), isopropenyl (-- c (ch₃)=ch₂)、 Vinyl and substituted vinyl etc..

" alkynyl " includes acetylene series unsaturated alkyl, in certain embodiments, has at most about 11 carbon atoms, it can To be straight or branched and there is at least one or 1 to 2 alkynyl unsaturation site.Alkynyl non-limiting Example includes acetylene series acetenyl, propargyl etc..

" aryl " includes phenyl, xenyl or naphthyl as the term is employed herein, and is preferably phenyl.Described Term includes substituted and unsubstituted part.Aryl can be replaced by any described part, including but do not limit In one or more parts selected from the group consisting of: halogen (fluorine, chlorine, bromine or iodine), alkyl, Hydroxyl, amino, alkyl amino, arylamino, alkoxyl, aryloxy group, nitro, cyano group, sulfonyl (sulfono), Sulfato base (sulfato), phosphono, phosphoric acid acyloxy or phosphonato, it is unprotected, or regards Need protected.

" cyclic alkyl " or cycloalkyl include 3-7 membered hydrocarbon ring, such as cyclopropyl.

" heterocycle " includes the 3-7 yuan of rings in ring with the carbon compound of 1 to 3 hetero atom (as o, s, n).

" aromatic heterocycle group " includes the aromatic ring containing 1 to 3 hetero atom (as o, s, n), such as pyridine radicals, Pyrimidine radicals.

" alkoxyl or alkyl oxy " includes group-or, and wherein r is alkyl.Specific alkoxyl include n-pentyloxy, Positive hexyloxy, 1,2- dimethyl butyrate epoxide etc..

" amino " includes group-nh₂.

Term " alkyl amino " or " arylamino " include thering is one or two alkyl or aryl substituent group respectively Amino.

" halogen " or " halogen " includes chlorine (cl), bromine (br), fluorine (f) or iodine (i).

" alkyl monosubstituted amino " includes group alkyl-nhr'-, and wherein r' is selected from alkyl or aryl.

" alkyl sulfenyl " includes group-sr, and wherein r is alkyl or aryl.

Unless otherwise defined, " protected " refers to for a group to be added to oxygen, nitrogen otherwise as the term is employed herein Or on phosphorus atoms, to prevent it from reacting further or for other purposes.Diversified oxygen and nitrogen-protecting group group It is that those of skill in the art in organic synthesis field are known.In the following methods the protection of functional group and go protect Shield can carry out (see, for example, t.w. Green by commonly known program in art And p.g.m. 5 this (p.g.m.wuts), (t.w.greene) " the blocking group in organic synthesiss (protecting groups in organic synthesis) ", the third edition, Willie publishing house (wiley), 1999), described document is incorporated herein by reference.Oxygen or " blocking group " of nitrogen Example include but is not limited to acyl group (such as acetyl group, formoxyl, benzoyl etc.), carbonate group (example As roc (o)-, wherein r can be substituted or unsubstituted alkyl, thiazolinyl, aryl, benzyl etc.), Carbamate groups (such as r_ar_bN-c (o)-, wherein r_aAnd r_bIt is each independently hydrogen, alkyl, aryl etc.). Oxygen and nitrogen-protecting group group can also include unsubstituted or substituted benzyl, pi-allyl, the tert-butyl group or first silicon Alkyl, it can easily be removed by known method in art.Especially, suitable nitrogen is protected Shield group illustrates as follows: benzyl-[bn], tert-butoxycarbonyl-[boc], t-butyldimethylsilyl - [tbdms] etc..

" leaving group " refers to that a group can be another by the such as reaction of displacement as the term is employed herein Group displacement.Suitable leaving group includes but is not limited to halogen (cl, br, i) and sulfonate group (- os (o)₂- Aryl (such as-os (o)₂Ph or-os (o)₂c₆h₄ch₃- p) or-os (o)₂- alkyl (such as-os (o)₂ch₃Or -os(o)₂cf₃)) etc..

" pharmaceutically acceptable salt " include its biological property of reservation of compound provided in this article and avirulence or Other any salt in accordance with the place of medicine use requirement invariably.

" prodrug " refers to when administration biosystem because spontaneous chemical reaction, enzyme catalysiss are anti-as the term is employed herein Should and/or metabolic process or respective combination and produce any compound of bioactive compound.Using connection To functional group (such as-oh ,-nh₂、-p(o)(nh)(oh)、-p(o)(oh)₂) and medicine associate, can be The group of internal cracking is forming Standard prodrugs.Heretofore described prodrug is exemplary, but is not only restricted to This, and one of ordinary skill in the art can prepare the prodrug of other Known Species.

Term " nucleoside " refers to purine or pyrimidine bases or its analog connecting to sugar, including its heterocycle and carbocyclic ring class Like thing.

Term " phosphate ester " refers to-o-po_32-

Term " phosphoramidate " refers to-n (r)-po_32-, wherein r is hydrogen or the substituent group based on carbon.

Term " biologically active drug or medicament " refers to produce the chemical entities of biological effect.In the present invention, biological Activating agent refers to nucleoside, single-nucleotide phosphate, nucleoside diphosphate ester, ribonucleoside triphosphote ester.

Term " alkaryl " or " alkylaryl " include the aryl with alkyl substituent.Term aralkyl or aryl Alkyl includes the alkyl with aryl substituent.

Term " deuterium " is the isotope of hydrogen, and atomic mass is 2 times of the latter, higher with the combination of carbon.Deuterate " and " deuterium " table Show that hydrogen is replaced by deuterium in specified location.One " substituent group of deuterate " is substituent group, and wherein at least one hydrogen is by refer to Fixed percentage ratio enrichment replaces deuterium.

Term " aminoacid " include the α that naturally occurs and synthesize-, β-, γ-or δ-aminoacid, and include But it is not limited to the aminoacid found in protein, i.e. glycine, alanine, L-Valine, leucine, different bright Propylhomoserin, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, Tyrosine, agedoite, L-Glutamine, aspartic acid, glutamic acid, lysine, arginine and histidine. In a preferred embodiment, aminoacid is in l- configuration or d- configuration.Or, aminoacid can be Following derivant: alanyl, valyl, leucyl-, isoleucyl-, prolyl, phenyl Alanyl, tryptophanyl, methionyl, glycyl, tryptophanyl, Threonyl, cysteinyl Base, tyrosyl-, asparaginyl-, glutaminyl, aspartyl, glutamyl, lysyl Base, arginyl-, histidyl-, β-alanyl, β-valyl, β-leucyl-, β-different bright Aminoacyl, β-prolyl, beta-phenyl alanyl, β-tryptophanyl, β-methionyl, β- Glycyl, β-tryptophanyl, β-Threonyl, β-cysteinyl-, β-tyrosyl-, β-sky Winter amide acyl group, β-glutaminyl, β-aspartyl, β-glutamyl, β-lysyl-, β-arginyl- or β-histidyl-.

" therapeutic agent " (therapeutic agent/therapeutic agents) refers to as the term is employed herein Can be used for treating or prevent any medicament of disease or one or more symptom.In certain embodiments, term " therapeutic agent " includes compound provided herein.In one embodiment, therapeutic agent is to be applied to known to one kind Or or currently be used for treating or prevent the medicament of disease or one or more symptom.

" therapeutically effective amount " includes right enough to realize when being used for treating disease to experimenter's administration compound or compositionss The amount of this treatment of described disease.

" treatment " (treating/treatment) of any disease or disease refers to that improvement is subject in one embodiment Disease in the presence of examination person or disease.In another embodiment, " treat " inclusion improve at least one can Can be difficult to be debated other body parameter by experimenter.In yet another embodiment, " treat " and include (example on body As other symptom stably can be debated) or physiologically (for example stablize body parameter) or adjust disease in terms of this is two Or disease.In yet another embodiment, " treat " outbreak including postponing disease or disease.

Present invention preparation is convenient, and product has antiviral effect.

The preparation of compound

Compound provided herein can obviously any method be prepared, is separated by one of ordinary skill in the art Or obtain.Illustrative preparation method is described in detail in following examples.

With reference to embodiment, the invention will be further described

Specific embodiment

(5r) -5- (((t-butyldimethylsilyl) epoxide) methyl) -4- hydroxyl dihydro -2 (3h) -one (11)

Compound 10 (39.4g, 300mmol) is dissolved in 120ml dmf, is cooled with an ice bath to zero degree, add in solution Enter imidazoles (51g, 0.75mol) and tert-butyldimethylsilyl chloride compound (49.5g, 0.33mol).Reactant mixture exists After being stirred overnight at room temperature, add water (150ml), stir ten minutes, extracted three times with ethyl acetate (250ml), merge organic Phase, washes three times with water (100ml).Separate organic layer, sodium sulfate is dried, decompression boils off solvent, residue silica gel column chromatography (hexane of 0-10% ethyl acetate) purification, obtains white solid product 11 (42.87 grams), yield 58%.

¹h nmr(400mhz,cdcl₃) δ 4.50-4.53 (m, 1h), 4.44 (s, 1h), 3.81 (t, j=2.8hz, 2h), 3.69 (d, j=4.4hz, 1h), 2.87 (dd, j=18hz, j=6.6hz, 1h), 2.42 (dd, j=18hz, j=1.6hz, 1h),0.85(s,9h),0.045(s,3h),0.03(s,3h).

¹³c nmr(101mhz,cdcl₃)δ177.24,88.25,69.52,63.05,38.93,25.74,22.75, 18.15,-5.64,-5.70.

(5r) -5- (((t-butyldimethylsilyl) epoxide) methyl) -4- hydroxyl -3,3- dimethyl dihydrofuran -2 (3h) -one (12)

The solution dry ice-propanone bath of 11 (22g, 89.3mmol) and thf (450ml) is cooled to -78 DEG C, slowly Deca diisopropylamine lithium reagent (tetrahydrofuran solution of 2m, 225ml, 0.45mmol, 5.0 equivalents), continues after adding Stir 50 minutes at a temperature of this, Deca iodomethane (31.7 grams, 223mmol, 2.5 equivalents).Mixture stirs at -78 DEG C Mix half an hour, be gradually heating to -30 DEG C, complete to reacting.It is cooled to -78 DEG C, add 200ml saturated ammonium chloride solution, After being gradually increased to room temperature, decompression boils off solvent, and residue adds ethyl acetate (600ml), washes three times with saline (100ml). Separate organic layer, sodium sulfate is dried, decompression boils off solvent, residue is with silica gel column chromatography (hexane of 0-40% ethyl acetate) Purification.Obtain white solid product 12 (14.95 grams), yield 61%.

¹h nmr(400mhz,cdcl₃)δ4.15(m,2h),3.88-3.97(m,2h),2.26(m,1h),1.28 (s,3h),1.21(s,3h),0.90(s,9h),0.1(s,3h),0.09(s,3h).

¹³c nmr(101mhz,cdcl₃)δ179.08,80.66,62.13,43.58,25.79,22.75,18.22, 17.96,-5.44,-5.48.

(5r) -4- (((t-butyldimethylsilyl) epoxide) -5- (((t-butyldimethylsilyl) epoxide) Methyl) -3,3- dimethyl dihydrofuran -2 (3h) -one (13)

12 (12 grams, 43.7mmol) are dissolved in 220ml dichloromethane, sequentially add imidazoles (7.31g, 108mmol) and the tert-butyl group Dimethylsilylchloride (9.8g, 65.3mmol).After reactant mixture is stirred at room temperature overnight, add methanol (15ml), Stirring ten minutes, removal of solvent under reduced pressure, residue adds ethyl acetate (300ml) and water (50ml).Separate organic layer, sulphuric acid Sodium is dried, and decompression boils off solvent.Residue passes through column chromatography (hexane solution of 0-20% ethyl acetate) purification, obtains 3 ', 5'- Oxygen-t-butyldimethylsilyl midbody product 13,15.47g, yield 91%.

(5r) -4- (((t-butyldimethylsilyl) epoxide) -5- (((t-butyldimethylsilyl) epoxide) Methyl) -3,3- dimethyl-tetrahydrofuran -2- methanesulfonates (15)

The mixture of 13 (1.2g, 3.09mmol) and thf (15ml) is cooled to 0 DEG C, adds li (obu-t)₃alh (6.2ml, 6.2mmol), subsequently reacts 40 minutes at 0 DEG C, is slowly increased to room temperature and continues stirring one hour.It is cooled to After 0 DEG C, add methanol that reaction is quenched.Decompression boils off solvent, and residue adds ethyl acetate (90ml) and water (10ml), Organic layer uses 1n hydrochloric acid solution (10ml), saturated sodium bicarbonate solution (20ml) and saturated aqueous common salt (30ml) to wash respectively Wash, anhydrous sodium sulfate drying, decompression boils off solvent.Add 15ml dichloromethane and be cooled to 0 DEG C in gained crude product 14 , add triethylamine (0.68ml, 5mmol), be slowly added dropwise mscl (0.31ml, 4.0mmol), be slowly increased to room temperature And continue to stir two hours.Add 60ml dichloromethane and 5% sodium bicarbonate (10ml) that reaction is quenched.Organic layer is used respectively Saturated sodium bicarbonate solution (10ml) and saturated aqueous common salt (20ml) washing, anhydrous sodium sulfate drying, decompression boils off solvent, The crude product 15 obtaining is directly used in next step reaction.

1- ((2r, 5r) -4- hydroxyl -5- (methylol) -3,3- dimethyl-tetrahydrofuran -2- base) pyrimidine -2,4 (1h, 3h) - Diketone (1001)

Under argon protection, add uracil (280mg, 2.5mmol), second cyanogen (15ml) and n, 0- in 50ml reaction bulb Double methylacetamides (1.27ml, 5.2mmol).It is warming up to 80 DEG C of stirrings 20 minutes to turbid solution clarification, reactant liquor is cold But to room temperature, compound 15 (938mg, 2.0mmol) and trimethylsilyl triflate (1.08ml, 6.0mmol) are successively added. Reactant liquor stirs 6 hours under 0 DEG C to room temperature, is quenched with saturated sodium bicarbonate after cooling, separates organic faciess, washing, is dried, Concentrate, residue 16 is dissolved in oxolane, add triethylamine hydrogen fluoride, be stirred at room temperature 2 days.After concentration, residue Purified (methylene chloride/methanol 30:1 to 20:1) with column chromatography, obtain white solid product 1001 (170mg, gross production rate 33%).

¹H nmr (400mhz, meod) δ 8.24 (d, j=8.1hz, 1h), 5.86 (s, 1h), 5.70 (d, j =8.1hz, 1h), 4.09 3.69 (m, 4h), 1.18 (s, 3h), 0.94 (s, 3h).

ms-esi(m/z)257.3(m+1).

Isopropyl [(s) (phenyl-pentafluoride epoxide) (phenoxy group) phosphoryl] l alanine ester (17)

Add dichloro-phenyl phosphate (11.42,51.4mmol) in reaction bulb, and l- alanine isopropyl ester hydrochlorate (8.64g, 51.4mmol), it is cooled to -78 DEG C, the dichloromethane (40ml) of Deca triethylamine (14.4ml, 103.5mmol) in 2 hours Solution, finishes, and is warmed to room temperature, and after being stirred overnight, reactant mixture is cooled to 0 DEG C, in 1 hour by Pentafluorophenol (9.46g, 51.4mmol) it is added drop-wise in above-mentioned solution, 0 with dichloromethane (50ml) solution of triethylamine (7.2ml, 51.4mmol) DEG C stirring 1 hour, be warmed to room temperature, after being stirred overnight, be filtered to remove triethylamine hydrochloride solid, solid filter cake dichloromethane (3x 20ml) washs, filtrate reduced in volume, and residue adds t-butyl methyl ether (150ml), filters gained white solid Washed with t-butyl methyl ether (3x 10ml).All filtrates concentrate after merging, and gained filter cake adds oneself of 20% ethyl acetate Alkane mixed solvent (100ml), after filtration, gained solid 10% sodium bicarbonate solution washs to filtrate to neutral, then washes, Gained solid is vacuum dried 30 hours at 55 DEG C, and dried solid adds 25ml ethyl acetate-pentane (1:5) mixed solvent, Stirring 1 hour, after filtering gained solid with ethyl acetate-pentane (1:5,2x 5ml) washing, with first time gained solid Merge, be dried to obtain title compound 17, purity is more than 99%.

¹h nmr(400mhz,cdcl₃) δ 7.37 (d, j=7.6hz, 2h), 7.34 7.19 (m, 3h), 5.18 4.98 (m, 1h), 4.29 4.10 (m, 1h), 4.07 3.97 (m, 1h), 1.48 (d, j=7.0hz, 3h), 1.27 (d, j=5.9hz, 3h), 1.26 (d, j=5.9hz, 3h).

³¹p nmr(162mhz,cdcl₃)δ-1.81.

Isopropyl [(r) (phenyl-pentafluoride epoxide) (phenoxy group) phosphoryl] d alanine ester (18)

Specific embodiment reference compound (17)

³¹p nmr(162mhz,cdcl₃)δ-1.81.

New penta (chlorine (phenoxy group) phosphoryl)-l- alanine ethyl ester (19)

Add dichloro-phenyl phosphate (11.42g, 51.4mmol) in reaction bulb, and l- alanine peopentyl ester hydrochlorate (10.02g, 51.4mmol), it is cooled to -78 DEG C, the dichloromethane (40ml) of Deca triethylamine (14.4ml, 103.5mmol) in 2 hours Solution, finishes, and is warmed to room temperature, and after being stirred overnight, is filtered to remove triethylamine hydrochloride solid, solid filter cake dichloromethane (3 X 20ml) washing, filtrate reduced in volume, residue column chromatography purifies (ethyl acetate/hexane, 0 to 40% ethyl acetate), Obtain colorless liquid product 19 (11.35g, yield 66.1%).

¹h nmr(400mhz,cdcl₃)δ7.40-7.36(m,2h),7.28-7.25(m,3h),4.27-4.23(m, 2h), 3.95-3.83 (m, 2h), 1.55and 1.53 (d, j=6.8hz, 3h), 0.96 (s, 9h).

³¹p nmr(162mhz,cdcl₃)δ9.09,8.73.

Isopropyl [(s) ((((2r, 5r) 5 (2,4 dioxy 3,4 dihydro-pyrimidin 1 (2h) base) 3- hydroxyl Base -4,4 dimethyl-tetrahydrofuran 2 base) methoxyl group) (phenoxy group) phosphoryl)] l alanine ester (101a1)

Add nucleoside 1001 (102mg, 0.4mmol) and the anhydrous thf of 2.0ml in 10ml reaction tube, mixture is in ice-water bath In be cooled to 0 DEG C.Deca tert-butyl group chloride Grignard reagents (the 1m thf solution of 0.42ml, 0.42mmol), reaction is mixed Compound stirs 30min at 0 DEG C, subsequently at 0 DEG C Deca phosphorus reagent 17 (190mg, 0.42mmol) in 1ml thf In solution.Obtained clarifying reaction solution is warming up to 25 DEG C, stirs 1 day.Add saturation nh₄Cl (3ml), stirs Mix 5 minutes, mixture is diluted with ethyl acetate (60ml).Separate organic faciess, aqueous layer with ethyl acetate (10ml) extracts. The organic layer water (10ml) of merging, saturation nahco₃(2x 10ml), saline (10ml) wash, and through na₂so₄It is dried. Decompression boils off solvent, and residue, via silica gel column chromatography (dichloromethane of 0-5% methanol) purification, obtains white solid product 101a1 (128mg), yield 61%).

¹H nmr (400mhz, meod) δ 7.76 (d, j=8.1hz, 1h), 7.39-7.42 (m, 2h), 7.28-7.30 (m, 2h), 7.21-7.25 (m, 1h), 5.87 (s, 1h), 5.62 (d, j=8.1hz, 1h), 4.95-4.99 (m, 1h),4.48-4.52(m,1h),3.35-4.40(m,1h),3.98-4.01(m,1h),3.90-3.96(m,1h),3.86 (d, j=8.8hz, 1h), 1.37 (d, j=8.0hz, 3h), 1.24 (s, 3h), 1.23 (s, 3h), 1.19 (s, 3h), 0.93(s,3h).

¹³c nmr(101mhz,meod)δ172.94,164.52,150.79,141.02,129.51,124.87,120.0, 100.89,92.29,80.70,74.44,68.82,65.12,50.28,45.13,20.63,20.54,20.25,19.69, 19.34,19.28.

³¹p nmr(162mhz,methanol-d₄)δ3.64；

ms-esi 526.3(m+1).

Isopropyl [(r) ((((2r, 5r) 5 (2,4 dioxy 3,4 dihydro-pyrimidin 1 (2h) base) 3- hydroxyl Base -4,4 dimethyl-tetrahydrofuran 2 base) methoxyl group) (phenoxy group) phosphoryl)] d alanine ester (102b2)

Add nucleoside 1001 (52mg, 0.2mmol) and the anhydrous thf of 1.0ml in 10ml reaction tube, mixture is in ice-water bath It is cooled to 0 DEG C.Deca tert-butyl group chloride Grignard reagents (the 1m thf solution of 0.24ml, 0.24mmol), reaction mixing Thing stirs 30min at 0 DEG C, subsequently at 0 DEG C Deca phosphorus reagent 18 (100mg, 0.22mmol) in 1ml thf Solution.Obtained clarifying reaction solution is warming up to 25 DEG C, stirs 1 day.Add saturation nh₄Cl (3ml), stirring 5 minutes, mixture is diluted with ethyl acetate (60ml).Separate organic faciess, aqueous layer with ethyl acetate (10ml) extracts.Close And organic layer water (10ml), saturation nahco₃(2x 10ml), saline (10ml) wash, and through na₂so₄It is dried. Decompression boils off solvent, and residue, via silica gel column chromatography (dichloromethane of 0-5% methanol) purification, obtains white solid product 102b2 (57mg), yield 54%.

¹H nmr (400mhz, meod) δ 7.71 (d, j=8.1hz, 1h), 7.40-7.36 (m, 2h), 7.29-7.18 (m, 3h), 5.85 (s, 1h), 5.62 (d, j=8.1hz, 1h), 4.98-4.93 (m, 1h), 4.53-4.49 (m, 1h), 3.36-4.31 (m, 1h), 3.98-3.95 (m, 1h), 3.91-3.87 (m, 1h), 3.80 (d, j=8.8hz, 1h), 1.34 (d, j=7.1hz, 3h), 1.21 (d, j=6.1hz, 3h), 1.20 (d, j=6.1hz, 3h), 1.16 (s, 3h), 0.87(s,3h).

³¹p nmr(162mhz,methanol-d₄)δ3.32；

ms-esi 526.3(m+1).

Neopentyl ((((2r, 5r) 5 (2,4 dioxy 3,4 dihydro-pyrimidin 1 (2h) base) 3- hydroxyl -4,4 Dimethyl-tetrahydrofuran 2 base) methoxyl group) (phenoxy group) phosphoryl) l alanine ester (104a)

1- ((2r, 5r) -4- ((t-Butyldimethylsilyl) epoxide) -5- (hydroxyl) -3,3- dimethyl-tetrahydrofuran -2- base) pyrimidine - 2,4 (1h, 3h)-diketone (20)

The mixture of 16 (780mg, 1.6mmol) and methanol (40ml) is cooled to 0 DEG C, adds p-methyl benzenesulfonic acid (39mg), Subsequently it is warmed to room temperature, react 2 hours (tlc tracking) at room temperature.Decompression boils off solvent, and residue adds ethyl acetate (90ml) With water (10ml), organic layer is washed with saturated sodium bicarbonate solution (20ml) and saturated aqueous common salt (30ml) respectively, anhydrous Sodium sulfate is dried, and decompression boils off solvent, and residue, via silica gel column chromatography (dichloromethane of 0-5% methanol) purification, obtains white Solid product 20 (530mg), yield 90%.

¹h nmr(400mhz,cdcl₃) δ 8.26 (d, j=8.1hz, 1h), 5.84 (s, 1h), 5.68 (d, j =8.1hz, 1h), 4.06 (d, j=8.4hz, 1h), 3.96 (dd, j=12.4,2.0hz, 1h), 3.82-3.79 (m, 1h), 3.72 (dd, j=12.4,2.4hz, 1h), 1.17 (s, 3h), 0.92 (s, 12h), 0.14 (s, 3h), 0.11(s,3h).

¹³c nmr(101mhz,meod)δ166.05,152.52,142.99,101.87,93.19,84.75,75.99,60.12, 47.08,26.29,21.83,21.41,18.84,-4.05,-4.32.

ms-esi 371.2(m+1).

Neopentyl ((((2r, 5r) 3 ((t-Butyldimethylsilyl) epoxide) 5 (2,4 dioxies 3,4 Dihydro-pyrimidin 1 (2h) base) 3- hydroxyl 4,4 dimethyl-tetrahydrofuran 2 base) methoxyl group) (phenoxy group) phosphorus Acyl group) l alanine ester (21)

Nucleoside 20 (80mg, 0.21mmol), phosphorus reagent (140mg, 0.42mmol) and 1.0ml is added in 10ml reaction tube Anhydrous thf, mixture adds the phonetic azoles of N-methyl at room temperature, and stirring completes for 3 hours to reaction.Add saturation nh₄Cl (3ml), Stirring 5 minutes, mixture is diluted with ethyl acetate (60ml).Separate organic faciess, aqueous layer with ethyl acetate (10ml) extracts. The organic layer water (10ml) of merging, saturation nahco₃(2x 10ml), saline (10ml) wash, and through na₂so₄It is dried. Decompression boils off solvent, and residue, via silica gel column chromatography (dichloromethane of 0-5% methanol) purification, obtains white solid product 21 (91mg), yield 65%.

¹h nmr(400mhz,cdcl₃) δ 9.25and 8.92 (s, 1h), 7.72and 7.64 (d, j=8.1hz, 1h),7.36-7.32(m,2h),7.44–7.06(m,6h),7.24-7.18(m,3h),5.91and 5.90(s, 1h), 5.66and 5.61 (d, j=8.1hz, 1h), 5.30 (s, 2h), 4.57 4.51 (m, 1h), 4.26 4.05 (m, 3h), 3.93 3.72 (m, 5h), 1.74 (s, 1h), 1.74 (s, 1h), 1.42 (d, j=7.0 hz,3h),1.18(s,3h),0.92-0.89(m,21h),0.10-0.05(m,6h).

³¹p nmr(162mhz,methanol-d₄)δ2.71,2.41；

ms-esi 668.3(m+1).

At 0 DEG C, in 25ml reaction bulb, add nucleoside prodrugs 21 (67mg, 0.1mmol) and 80% trifluoroacetic acid aqueous solution, Mixture is warmed to room temperature, and reacts 3 hours (tlc tracking) at room temperature, and decompression boils off solvent, and residue adds ethyl acetate (90ml) with water (10ml), organic layer is washed with saturated sodium bicarbonate solution (20ml) and saturated aqueous common salt (30ml) respectively, Anhydrous sodium sulfate drying, decompression boils off solvent, and residue, via silica gel column chromatography (dichloromethane of 0-5% methanol) purification, obtains White solid product 104a (50mg), yield 90%.

¹h nmr(400mhz,methanol-d₄) δ 7.77and 7.72 (d, j=8.1hz, 1h), 7.40-7.18 (m, 5h), 5.86and 5.85 (s, 1h), 5.61and 5.60 (d, j=8.1hz, 1h), 4.56-4.35 (m, 2h), 4.03-3.96 (m, 2h), 3.87-3.73 (m, 3h), 1.39and 1.36 (d, j=7.2hz, 3h), 1.18 (s, 3h),0.94-0.89(m,12h).

³¹p nmr(162mhz,methanol-d₄)δ3.67,3.56；

ms-esi 554.2(m+1).

Hcv replicon is analyzed

Anti- hcv activity and the toxicity of Exemplary compounds can be tested in following two bioanalysiss: based on cell The analysis of hcv replicon and cytotoxicity analysis (wo2007/027248).

I. anti-hcv analysis

Using the human liver cancer adding luciferase reporter gene (luc-ubi-neo) containing duplication hcv two sub-gene group genotype 1b replicon Cell line (huh-7) is active come the anti-hcv to assess compound.In this analysis, luciferase signal level and virus rna Replicate directly related.It is being supplemented with 10% hyclone and 0.5mg/ml Geneticin (geneticin) hcv replicon-reporter cell lines (nk/luc-ubi-neo) are cultivated in the dmem culture medium of (g418). Cell is made to maintain subconfluent (subconfluent) state to guarantee high-caliber hcv replicon rna synthesis. For assessing the antiviral activity of compound, prepare serial dilution in 0.10 to 300 μ m for the concentration.Change by dilution Compound is transferred in 96 orifice plates, then adds replicon cell (6000 cells in every hole).Cell is trained together with compound Educate 48h, measure uciferase activity after this.Luciferase signal weakens and is reflected in hcv replicon rna in processed cell Reduce and be used for determine ec50 value (making uciferase activity reduce by 50% concentration).

Ii. cytotoxicity analysis

Luciferase reporter gene (by hiv ltr promoters driven) is carried using being stably integrated in chromosome The cytotoxic effect to analyze selected compounds for the huh-7 cell line.Make this cell line (ltr-luc) maintain to have In the dmem culture medium of 10%fbs.The design of cytotoxicity analysis is similar to the design of hcv replicon analysis.Processed thin In born of the same parents, the reduction of uciferase activity and the cytotoxic effect of test compound are related and be used for calculating cc50 value (suppression is thin Intracellular growth reaches 50% concentration).

The biological results of selected compounds are summarized in table 1.

Activity in replicon analysis (genotype 1b) for table 1. Exemplary compounds

Ec in hcv-repl luc algoscopy is provided below₅₀:

0.1 μm≤+++ +≤0.5 μm≤+++≤1 μm≤++≤10 μm of ＜+

Cc in hcv-repl luc algoscopy is provided below₅₀:

1 μm≤++≤10 μm of ＜+

Ec50 is the drug level that suppression hcv reaches 50%.

Cc50 reaches 50% drug level for cell growth inhibiting.

The result of the anti-hcv activity of the selected nucleoside prodrugs summarized in table 1 shows that these nucleoside phosphorylase ester prodrugs show the anti-hcv of effect Activity, and no notable cytotoxicity, which ensure that to research and development as the novel nucleoside disclosed herein of anti-hcv agent or its prodrug Study further.

The previous examples of preferred embodiment and description are interpreted as illustrative, rather than limit as by claims institute

The present invention defining.As will readily appreciate that, can be in the feelings without departing substantially from the present invention as disclosed in the claims Utilize many changes of features described above under condition and combine.All such changes are intended to be included in the range of claims.

Claims

1. the nucleoside analog that one kind 2 '-bis- alkyl replace, is characterized in that: is nucleoside represented by formula i, nucleoside ammonia Base phosphate compound or its pharmaceutically acceptable prodrug, salt, solvate, stereoisomer, change Isomer or polymorphic forms, metabolite:

Wherein,

(1) base is natural or engineered miazines or purines nucleoside base；

(4) w is oxygen or sulfur；

(5) x and y is each independent, and one of is-h ,-or₁、-sr₁、-nr₁r₂, with n connect natural or Non-natural amino acid estersOr the natural or non-natural amino acid esters being connected with oxygen；And another is then For-or₁,-nr₁r₂；

2. the nucleoside analog that according to claim 12 '-bis- alkyl replace, is characterized in that: selected from followingization Compound ii to viii:

Wherein,

(1) base is selected from

Or its tautomer

Wherein r₃For h, oh, nh₂,sme,or’,f,cl,br,i,-n₃,nhoh,-nhor’,-nhr”；

r₄For h, oh, or ', sr ', nhr ", f, cl, br, i ,-n₃,nhoh,-nhor’；

r₅For h, oh, nh₂,or’,nhr”,f,cl,br,i,nhoh,-nhor’；

r₇For h, d, oh, or ', sr ', nhr ", f, cl, br, i, cn, n₃；

(3)w、x、y、z、r、r₁And r₂As defined in claim 1；

3. the nucleoside analog that according to claim 22 '-bis- alkyl replace, is characterized in that: selected from Lower structural formula ix to xiii:

Wherein,

Hal is halogen, including fluorine, chlorine, bromine, iodine；Other substituent groups w, x, y, z, base such as right Require defined in 1.

4. the nucleoside analog that 2 ' according to any one of claim 1-3-bis- alkyl replace, its feature : the hydrogen atom on optional position in the molecule shown in general structure i to xiii can be replaced by deuterium.

5. the nucleoside analog that 2 ' any one of claim 1-4-bis- alkyl replace, is characterized in that: The following oxide precursor 1001-1084 of selection:

6. the nucleoside analog that 2 ' any one of according to claim 1-5-bis- alkyl replace, it is special Levying is: the following compound 101-136,201-233,301-337,401-434 of selection:

7. the nucleoside analog that 2 ' according to any one of claim 1-5-bis- alkyl replace, is characterized in that: The following compound 501-665 of selection:

8. the nucleoside analog that 2 ' according to any one of claim 1-7-bis- alkyl replace, its feature It is: the 2 '-phosphate prodrugs of nucleoside compound of bis- alkyl replacements, officinal salt, hydrate, solvate Or crystallization is prepared by mixing into pharmaceutical preparation and nanometer system with pharmaceutically acceptable carrier, diluent or excipient Agent, to be suitable for oral or parenteral；Medication includes, Intradermal, intramuscular, intraperitoneal, intravenouss, Subcutaneous, intranasal and peroral route；Described preparation can be applied by any approach, for example, pass through infusion or inject, Applied by the approach of transepithelial or mucocutaneous absorption；Administration can be whole body or local；Oral administration The example of preparation includes solid or liquid dosage form, specifically, including tablet, pill, granule, powder, glue Wafer, syrup, Emulsion, suspensoid；Described preparation can be prepared by methods known in the art, and comprises medicine The conventional use of carrier of thing formulation art, diluent or excipient, nanometer formulation；Officinal salt, hydrate, Solvate, crystallization and medically acceptable excipient；The group selected from consisting of of therapeutic agent independence and medicine Thing: ribavirin, interferon, hcvns3 protease inhibitor, the suppression of hcv reverse transcriptase ns5b non-nucleoside Agent, the synergist of hcv reverse transcriptase ns5b nucleosidic inhibitors, ns5a inhibitor and ns5a inhibitor, Entry inhibitor, ciclosporin immunosuppressant, ns4a antagonist, ns4b inhibitor, cyclophilin inhibitor.

9. the nucleoside analog that 2 ' according to any one of claim 1-8-bis- alkyl replace is anti-in preparation Application in the medicine of virus, is characterized in that: described virus includes hcv, hbv, hiv, rsv, norovirus, Ev71, dengue fever, influenza virus, Ebola virus, flaviviridae.

10. the nucleoside analog that according to claim 92 '-bis- alkyl replace is preparing the medicine of antiviral Application in thing, is characterized in that: described flaviviridae is hepatitis C viruss.