CN101240001B - 2'-fluoro nucleotide - Google Patents

Abstract

The present invention provides a 2'-fluor-nucleoside compound used for curing hepatitis b infection, hepatitis c infection, HIV and abnormal cells proliferation including tumour and cancer. The compound has formulas (I), (II), (III), (IV) or its medicine salt, wherein, base is purine or pyrimidine base; R<1> is OH, H, OR<3>, N<3>, CN, halogen includes F, or CF3, lower alkyl, amino, lower alkyl amino, two lower alkyl amino, or alkoxy; R<2> is H, phosphate, including monophosphate, diphosphate, triphosphate or stable phosphate prodrug; acyl, or other medical leaving group, when human body uses, the group provides a compound which R<2> is H or phosphate; sulfonic ester includes alkyl or aralkyl sulfonyl, including methyl sulphonyl, benzyl, wherein, benzyl is selectively substituted by the substituent of one or more aryl definition, lipid, amino, peptide or cholesterin; and R<3> is acyl, alkyl, phosphate or other medical leaving group, it is a group cracked by the parent compound when the human body uses.

Description

2'-fluoro nucleosides

The application is that international filing date is February 25, international application no in 1999 the dividing an application for the application of " 2 '-fluoro nucleosides " that be PCT/US99/04051 (national applications number be 99805472.0), denomination of invention.

The present invention who describes herein supports that by government the grant number that health ministry is authorized is AI32351.United States Government has confirmed right of the present invention.

Technical field

The invention belongs to the pharmaceutical chemistry field, comprise specifically 2 '-the fluoro nucleosides and its production and use.

Background technology

For some years, synthetic nucleosides such as 5-iodo-2 '-deoxyuridine and 5-fluoro-2 '-deoxyuridine has been used for the treatment of cancer and simplexvirus.Since the eighties in 20th century, synthetic nucleosides also has been to be conducive to treat HIV, hepatitis and Epstein-Barr virus.

In 1981, acquired immune deficiency syndrome (AIDS) (AIDS) was confirmed to be a kind of disease of serious harm human immune system, and it almost causes death without any exception.In nineteen eighty-three, the cause of disease of AIDS is defined as human immunodeficiency virus (HIV).In 1985, have report claim synthetic nucleosides 3 '-azido--3 '-the copying of deoxythymidine (AZT) HIV (human immunodeficiency virus) inhibiting.From that time, proved some other synthetic nucleosides, comprise 2 ', 3 '-didanosine (DDI), 2 ', 3 '-zalcitabine (DDC) and 2 ', 3 '-dideoxy-2 ', 3 '-two dehydrogenation thymidines (D4T), effective to HIV.By cell kinase by the cells phosphorylation effect produce 5 '-triphosphate after, these synthetic nucleosides mix in the growing chain of viral DNA, owing to do not have 3 '-hydroxyl causes chain termination.They also can suppress viral reverse transcriptase.

The success that multiple synthetic nucleosides suppresses HIV in vivo or has in vitro made some investigators' designs and has detected nucleosides, its 3 of nucleosides '-position replaces heteroatoms with carbon atom.European patent application No.0337713 and U.S. Patent No. 5,041,449 (authorizing BioChem Pharma, Inc.) disclose racemize 2-replacement-4-and have replaced-1,3-dioxolane, and it shows antiviral activity.U.S. Patent No. 5,047,407 and european patent application No.0382526 (also be to belong to BioChcmPharma, Inc.), disclose some and had racemic 2-replacement-5-replacement-1, the 3-oxa-thiacyclopentane nucleosides of antiviral activity, and particularly pointed out 2-methylol-5-(cytosine(Cyt)-1-yl)-1,3-oxa-thiacyclopentane (hereinafter being called BCH-189) has intimate identical anti-HIV activity with AZT, and toxicity is very little.(-) of racemic mixture BCH-189-enantiomer is called 3TC, and its U.S. Patent No. 5,539,116 by Liotta etc. is open, sells at present itself and AZT combination therapy HIV in the U.S..

Also disclose suitable-2-methylol-5-(5-flurocytosine-1-yl)-1,3-oxa-thiacyclopentane (" FTC ") has strong HIV (human immunodeficiency virus)-resistant activity.Schinazi etc., racemic mixture and the enantioselectivity HIV inhibiting of cytosine(Cyt) " with the suitable-5-fluoro-1-[2-methylol-1; 3-oxa-thiacyclopentane-5-yl] " Antimicrobial Agents and Chemotherapy, in November, 1992, pp.2423-2431.Also visible U.S. Patent No. 5,210,085; WO 91/11186 and WO 92/14743.

Other virus that causes people's serious health problem is hepatitis B virus (hereinafter being called " HBV ").HBV is the reason that causes human cancer that is only second to smoking.The mechanism that HBV brings out cancer it be unclear that.It directly causes the deterioration of tumour by inference, or indirectly causes the deterioration of tumour by chronic inflammatory diseases, liver cirrhosis and the cell regeneration relevant with infection.

After 2 to 6 months latent period, this moment, infected object was not yet discovered infection, and HBV infects can cause hepatitis and liver injury, and it causes the Enrichment of some enzyme in abdominal pain, jaundice and the blood.HBV can cause fulminant hepatitis, and this is that a kind of quick deterioration often causes dead disease, and most of liver is destroyed when suffering from this disease.

In general, the acute hepatitis patient can rehabilitation.But in some patients, the virus antigen of high density is long-term or be retained in indefinitely in the blood, causes chronic infection.Chronic infection can cause chronic persistent hepatitis.Chronic persistent HBV patient is the most general in developing country.In year in 1991, the 2.25 hundred million Chronic HBV carrier that only just have an appointment in the Asia, and there are nearly 300,000,000 carrier in the whole world.Chronic persistent hepatitis can cause fatigue, liver cirrhosis and hepatocellular carcinoma, a kind of main liver cancer.

In west industrialized country, the crowd that the HBV infection risk is high comprises those people that contact with HBV carrier or its blood sample.The epidemiology of HBV is similar to acquired immune deficiency syndrome (AIDS), the reason of common HBV infection in HIV infection or AIDS patient that Here it is.But HBV is than the easier contact infection of HIV.

It is active that FTC and 3TC show anti-HBV.Furman etc., " suitable-5-fluoro-1-[2-(methylol)-1; 3-oxa-thiacyclopentane-5-yl]-resistance of hepatitis B of (-) and (+) enantiomer of cytosine(Cyt) is active, cytotoxicity and metabolic characteristic " Antimicrobial Agents andChemotherapy, in December, 1992, pp.2686-2692; And Cheng etc., Journal ofbiological Chemistry, Volume 267 (20), pp.13938-13942 (1992).

Developed the vaccine that derives from human serum the patient has been carried out immunity with anti-HBV.Although having found it is that effectively the production of this vaccine has some troubles, this is because obtaining human serum from chronic carrier is restricted, and purification step is long and cost is high.In addition, the every batch of vaccine for preparing from different serum must detect to guarantee safety in chimpanzee.Also by the preparation vaccine.Also be likely with a kind of engineered protein alpha-interferon therapy every day.

Hepatitis C virus (" HCV ") is major cause (Alter, H.J. (1990) J.Gastro.Hepatol.1:78-94 of blood transfusion postoperative infection and accidental non-A non-B hepatitis; Dienstag, J.L. (1983) Gastro 85:439-462).Although improved screening methods, be still the cause of disease of at least 25% acute viral hepatitis (ibid for Alter, H.J. (1990) at a lot of national HCV; Dienstag, J.L. (1983), ibid; Alter M.J. etc., (1990a) J.A.M.A.264:2231-2235; Alter M.J. etc., (1992) N.Engl.J.Med.M:1899-1905; Alter, M.J. etc., (1990b) N.Engl.J.Med.321:1494-1500).With latent at high proportion, they may not show clinical symptom in a lot of years in chronic infection (and infectious) carrier in the HCV infection.Acute infection changes at high proportion to chronic infection (70-100%) and hepatopathy (＞50%), and it distributes and the shortage of vaccine widely, and making HCV is morbidity and dead remarkable reason.

Tumour is irregular, the histoclastic propagation of Growth of Cells.If it has infectivity and transitivity, then tumour is pernicious or carcinous.Infectivity refers to that tumour enters surrounding tissue, destroys the basic unit that limits organizational boundary, so enter the tendency of normal systemic circulatory system.Transitivity refers to tumor migration to other zone of health and in the former tendency that in addition foundation propagation zone, site occurs.

Cancer is that the present U.S. causes second dead reason.Made a definite diagnosis existing 8,000,000 people of the U.S. and to be suffered from cancer, estimate newly to diagnose out 1,208,000 in 1994.Die from 500,000 people that surpass of this disease every year in this country.

At molecular level, people do not fully realize cancer.Known cells contacting carcinogenic substance such as some virus, some chemical substance or radiation cause DNA to change this deactivation " inhibition " gene or activation " oncogene ".Suppressor gene is growth regulatory gene, after its sudden change, or else can control Growth of Cells.It is normal gene (being called proto-oncogene) that oncogene begins, and its change by sudden change or expressed sequence becomes transforming gene.The product of transforming gene causes unsuitable Growth of Cells.More than 20 kind of normal cell gene of change by gene can become oncogene.It is much different that the cell that has transformed and normal cell have, and comprises cellular form, cell-with-cell interaction, film forms, cytoskeletal structure, protein excretion, genetic expression and death (cell that has transformed can indeterminate growth).

The cell type that all of body are different can change optimum or malignant cell into.Modal tumor sites is lung, then is colorectum, breast, prostate gland, bladder, pancreas, besides ovary.Other more general type of cancer comprises leukemia, and central nervous system cancer comprises brain tumor, melanoma, lymphoma, erythroleukemia, uterus carcinoma and head and neck cancer.

Now cancer mainly one of with the following method or its cooperative programs treatment 3 years: operation, radiotherapy and chemotherapy.Operation comprises removes most of illing tissues.Although sometimes performing the operation removing the tumour that is positioned at some site such as breast, rectum and skin is effectively, it can not be used for the treatment of and is positioned at other zone such as the tumour of vertebra, can not be used for the treatment of tumour illness such as the leukemia of disseminating.

Chemotherapy comprises disintegrates copying or cellular metabolism of cell.Its most frequently used leukemia and mammary gland, lung and testis cancer for the treatment of.

The chemotherapeutics that is used for the treatment of at present cancer has 5 main kinds: natural product and derivative thereof; Anthracycline; Alkylating agent; Antiproliferative agents (being also referred to as antimetabolite); And hormone.Chemotherapeutics often is called antineoplastic agent.

It is believed that alkylating agent by with other possible base alkylation or crosslinked working among guanine and the DNA, stops cell fission.Typical alkylating agent comprises mustargen, ethylenimine compound, alkyl sulfuric ester, cis-platinum and multiple nitrosourea.The shortcoming of these compounds is that they not only connect malignant cell, also connects the cell that other divides naturally, such as the cell of marrow, skin, gastrointestinal mucosa and placenta tissue.

Antimetabolite generally is reversible or irreversible enzyme inhibitors, or the interfere RNA compound that copies, translate or transcribe.

Determined that some synthetic nucleosides show the anticancer activity.The nucleoside derivates of knowing with strong antitumour activity is 5 FU 5 fluorouracil.5 FU 5 fluorouracil has been used for the clinical treatment malignant tumour, for example, comprises cancer, sarcoma, skin carcinoma, Alimentary cancer and mammary cancer.But 5 FU 5 fluorouracil causes that serious side reaction is such as nauseating, alopecia, diarrhoea, stomatitis, leukocytic thrombocytopenia, apocleisis, pigmentation and oedema.Derivative with 5 FU 5 fluorouracil of antitumour activity is described in US Patent No 4,336,381, and Japanese patent application Nos.50-50383,50-50384,50-64281,51-146482 and 53-84981.

U.S. Patent No. 4,000,137 have described inosine, adenosine or cytidine and methyl alcohol or ethanol has the leukemic activity of antilymphocyte through the snperoxiaized product of superoxide.

Cytosine arabinoside (being also referred to as Cytarabin, araC and Cytosar) is a kind of nucleoside analog of deoxidation cytosine(Cyt), its at first in nineteen fifty synthetic and in 1963 as clinical medicine.It is the most important medicine for the treatment of at present acute myeloid leukemia.It also has activity to acute lymphoblastic leukemia, and less being used for the treatment of in chronic myelocytic leukemia and the non_hodgkin lymphoma.The Main Function of cytosine arabinoside is suppress nucleic acid DNA synthetic.Handschumacher, R. and Cheng, Y., " purine and pyrimidine antimetabolite ", and Cancer Medicine, the XV-1 chapter, the 3rd edition, editor J.Holland etc., Lea and Febigol publish.

5-azacitidine belongs to the cytidine class, and it is mainly used in treating acute myelocytic leukemia and myelodysplastic syndrome.

2-fluorine adenosine-5 '-phosphoric acid salt (Fludara is also referred to as FaraA) is one of the most effective medicine for the treatment of chronic lymphocytic leukemia.DNA is synthetic to work this compound by suppressing.With F-araA process cell with at GI/S phase edge and relevant at the cellular accumulation of S phase; Therefore, it is the specific drugs of cell cycle S phase.Active metabolite F-araATP mixes, and has stoped the extension of DAN chain.F-araA or the potent inhibitor of ribonucleotide reductase, this enzyme are the key enzymes of being responsible for forming dATP.

2-chlorodeoxyadenosine is used for the treatment of rudimentary B glucagonoma such as chronic lymphocytic leukemia, non_hodgkin lymphoma and hairy cell leukemia.

When designing new biological activity nucleosides, people several times attempt fluoro substituents is mixed in the carbohydrate ring of nucleosides.Fluorine suggestion is substituting group, be because it may as hydroxyl wait the utmost point and isopyknic stand-in because C-F bond distance's (1.35 dust) is very approaching with C-O bond distance's (1.43 dust), and because fluorine is the acceptor of hydrogen bond.Fluorine can make molecule produce significant electronic variable with minimum spatial interference.Replace the variation that other group in the molecule can cause substrate utilization with fluorine, this is (116kcal/mol and C-H=100kcal/mol) because the high strength of C-F key.

Some reference have reported 2 '-the synthetic and purposes of arabinose base fluoro nucleosides (that is, 2 '-fluorin radical is the nucleosides of " making progress " configuration).Existing some reports have reported that 2-fluoro-beta-D-furyl glycosyl nucleosides shows the activity of resistance of hepatitis B and bleb.For example, referring to U.S. Patent No. 4,666,892 (Fox etc.); U.S. Patent No. 4,211,773 (Lopez etc.); Su etc., Nucleosides.136, " the synthetic and antivirus action of several 1-(2-deoxidation-2-fluoro-beta-D-furyl glycosyl)-5-alkyl urea pyrimidine "; " some structure activity relationships ", J.Med Chem., 1986,29,151-154; Borthwick etc., " carbocyclic ring 2 '-arabinose-fluoro-guanosine: a kind of strong synthesizing with enzyme of new herpes agent splits, " J.Chem.Soc., Chem.Commun, 1988; Wantanabe etc., " active anti-AIDS nucleosides 3 '-azido--3 '-deoxythymidine (AZT) and 2 '; 3 '-zalcitabine (DDC) 2 '-" go up " fluorine analogue synthetic and HIV (human immunodeficiency virus)-resistant activity ", J.Med.Chem.1990.33,2145-2150; Martin etc., " the synthetic and antiviral activity of single fluorine of the pyrimidine dideoxy ribonucleoside of anti-human immunodeficiency virus (HIV-1) and two fluoro analogues, " J Med, Chem.1990,33,2137-2145; Sterzycki etc., " some 2 '-the synthetic and HIV (human immunodeficiency virus)-resistant activity of fluoro-contain pyrimidine nucleoside, " J. Med.Chem.1990, and EPA 0316017 (by applications such as Sterzycki); And Montgomery etc., " 9-(2-deoxidation-2-fluoro-beta-D-furyl glycosyl) guanosine: 2 of a class metabolic stability '-pancreatic desoxyribonuclease cell toxicant analogue ", U.S. Patent No. 5,246,924 disclose the method for the treatment of hepatitis, comprise use 1-(2 '-deoxidation-2 '-fluoro-beta-D-furyl glycosyl)-the 3-ethyl uracil), be also referred to as " FEAU ".U.S. Patent No. 5,034,518 disclose 2-fluoro-9-(2-deoxidation-2-fluoro-beta-D-furyl glycosyl) adenosine class, and it shows antitumour activity by reducing compound as the metabolism that the ability of thymidine substrate changes adenosine.EPA 0292023 discloses some β-D-2 '-fluoro arabinose yl nucleosides has anti-virus infection active.

U.S. Patent No. 5,128,458 disclose antiviral agent β-D-2 ', 3 '-dideoxy-4 '-the thioribose nucleosides.U.S. Patent No. 5,446,029 disclose have 2 of antihepatitic activity ', 3 '-dideoxy-3 '-the fluoro nucleosides.

European patent application No.0409227 A2 disclose be used for the treatment of hepatitis B some 3 '-β that replaces-D-pyrimidine and purine nucleoside.

Also disclose L-FMAU (2 '-fluoro-5-methyl-β-L-furyl glycosyl uridylic) and be a kind of strong anti--HBV and anti--EBV agent.See Chu etc., " 2 '-fluoro-5-methyl-β-L-furyl glycosyl uridylic as the purposes of new antiviral agent treatment hepatitis B virus and Epstein-Barr virus " AntimicrobialAgents and Chemotherapy, April nineteen ninety-five, 979-981 page or leaf; Balakrishna etc., " with new L-nucleosides, 2 '-fluoro-5-methyl-β-L-furyl glycosyl uridylic inhibition hepatitis B virus ", Antimicrobial Agents and Chemotherapy, in February, 1996,380-356 page or leaf; United States Patent(USP) Nos. 5,587,362; 5,567,688; And 5,565,438.

United States Patent(USP) Nos. 5,426,183 and 5,424,416 disclose preparation 2 '-deoxidation-2 ', 2 '-two fluoro nucleosides and 2 '-deoxidation-2 '-method of fluoro nucleosides.Also can referring to " 2 ', 2 '-people's deoxycytidine kinase of difluoro Deoxyribose cytidine (gemcitabine) and purifying and the dynamics research of cytidine deaminase ", BioChemical Pharmacology, Vol.45 (No.9) 4857-1861 page or leaf, 1993.

U.S. Patent No. 5,446,029 (Eriksson etc.) disclose have the resistance of hepatitis B activity some 2 ', 3 '-dideoxy-3 '-the fluoro nucleosides.U.S. Patent No. 5,128,458 disclose some 2 ', 3 '-dideoxy-4 '-thionucleoside, wherein 3 '-substituting group is H, trinitride or fluorine.WO 94/14831 disclose some 3 '-fluoro-dihydro-pyrimidin nucleosides.WO 92/08727 disclose β-L-2 '-deoxidation-3 '-uridine nucleosides that fluoro-5-replaces, be used for the treatment of herpes simplex 1 and 2.

EPA publication number No.0352248 discloses the L-ribofuranosylpurine nucleosides that a large class is used for the treatment of HIV, bleb and hepatitis.Although some 2 '-the fluoro purine nucleoside belongs in this large class, do not provide the information of these compounds of preparation at its specification sheets, and their preferred row of not listing in open especially or specification sheets.How this specification sheets open does not prepare 3 if not being '-ribofuranosyl fluoridizes nucleosides.Similarly explanation is found in WO 88/09001 (Aktiebolaget Astra application).

European patent application 0357571 discloses β-D and the α-D pyrimidine nucleoside that a large group is used for the treatment of AIDS, in this large group, comprise in general manner 2 ' or 3 ' position nucleosides that can be replaced by fluorin radical.But, in this large group, do not disclose especially 2 '-fluoro nucleosides or its preparation method.

The open preparation of EPA 0463470 (5S)-3-fluoro-tetrahydrochysene-5[(hydroxyl) methyl]-preparation method of 2-(3H)-furanone, its be the preparation 2 '-fluoro-2 ', 3 '-di-deoxynucleoside such as 2 '-fluoro-2 ', 3 '-a kind of known intermediate of zalcitabine.

U.S.S.N.07/556,713 disclose β-D-2 '-fluorine furyl glycosyl nucleosides, and preparation method thereof, they are synthetic 2 ', 3 '-dideoxy-2 '-intermediate of fluorine arabinose yl nucleosides.

U.S. Patent No. 4,625,020 discloses by the method for 1,3,5-, three-O-acyl group-ribofuranose preparation with the 1-halo of protecting group-2-deoxidation-2-fluorine furyl glycosyl derivative.

The open very shortage that comprises β-L-2 ' of treat HIV, hepatitis (B-mode or the third type) or proliferative disorders-fluoro-ribofuranose yl nucleosides to being used for medical use.At least relevant 2 '-the ribofuranose yl nucleosides, this may be since with presensation fluorin radical is placed 2 '-the ribofuranosyl configuration is difficult.For L-2 '-fluoro-2 ', 3 '-unsaturated purine nucleoside, may be since purine nucleoside unstable in acidic medium, cause the fracture of glucose base key.

Consider the HIV acquired immune deficiency syndrome (AIDS), AIDS-related syndromes and hepatitis B and hepatitis C virus are at worldwide dissemination, and the patient who infects caused the fact of miserable impact, also exist for treat these diseases provide new, effectively, to the requirement of the medicament of the object low toxicity for the treatment of.In addition, the demand that also has the antiproliferative that provides new.

Summary of the invention

Therefore, one of purpose of the present invention provides the method and composition that the patient of hepatitis B or hepatitis C has been infected in treatment.

Another object of the present invention provides the method and composition that the patient of HIV has been infected in treatment.

Another object of the present invention provides new antiproliferative.

Another object of the present invention provide preparation 2 '-novel method of fluoro-ribofuranose yl nucleosides.

Another object of the present invention provide preparation 2 ', 3 '-dideoxy-2 ', 3 '-two dehydrogenations-2 '-novel method of fluoro-L-glyceryl-penta-2-thiazolinyl-furyl glycosyl nucleosides.

Specific embodiments

In one embodiment of the invention, provide structure following 2 '-α-fluoro-nucleosides:

Wherein

Base is purine or the pyrimidine bases that further define as herein;

R ¹OH, H, OR ³, N ₃, CN, halogen comprise F or CF ₃, low alkyl group, amino, low-grade alkyl amino, two elementary alkyl amido or alkoxyl group, and base refers to purine or pyrimidine bases;

R ²That H, phosphate radical comprise the monophosphate root, gen-diphosphate, triphosphate, or stable phosphate prodrug; Acyl group, or other medicinal leavings group, this group can provide R when using in the body ²It is the compound of H or phosphate radical; Sulphonate comprises alkyl or aralkyl alkylsulfonyl (comprising methyl sulphonyl), benzyl (wherein the phenyl selectivity is replaced by the substituting group described in one or more above-mentioned aryl definition), lipid (comprising phosphatide), amino acid, peptide or cholesterol; And

R ³Be acyl group, alkyl, phosphate radical or other medicinal leavings group, namely when using in the body, can be cracked into the group of parent compound.

In second embodiment, provide 2 '-fluoro nucleosides of following formula:

Wherein substituting group as defined above.

In the 3rd embodiment, provide 2 '-fluoro nucleosides of following formula:

Wherein substituting group as defined above.

In the 4th embodiment, provide structure following 2 '-fluoro nucleosides:

Wherein substituting group as defined above.

These 2 '-fluoro nucleosides can be β-L or β-D configuration.Preferred β-L configuration.

This type of 2 '-fluoro nucleosides is bioactive molecules, and they are used for the treatment of hepatitis B, hepatitis C or HIV.These compounds also are used for the treatment of abnormal cell proliferation, comprise tumour and cancer.Estimate this compound by the detection method of description herein or with other affirmation detection method, can easily determine activity profile.

In another embodiment, in order to treat hepatitis or HIV, this active compound or derivatives thereof or salt can or replace administration with other antiviral agent (for example anti-hiv agent or anti-hepatitis agent comprise that as above structural formula is described) associating.In general, in combination therapy, the together administration of two or more reagent of significant quantity, and in alternating treatment, all ingredients sequential administration of significant quantity.Dosage depends on absorption, deactivation and drainage rate and the other factors well known by persons skilled in the art of medicine.It should be noted that this dosage also will change with the seriousness of the illness that is treated.It should also be further understood that for any specific object specific dosage and program are according to needs and the administration person of individuality or instruct the people of these composition administrations to rely on professional judgement to be adjusted in time.

Can comprise 2-methylol-5-(5-flurocytosine-1-yl)-1,3-oxa-thiacyclopentane (FTC) with the limiting examples of the antiviral agent of compound disclosed herein associating; 2-methylol-5-(cytosine(Cyt)-1-yl)-1, (-)-enantiomer of 3-oxa-thiacyclopentane (3TC); Carbovir, acyclovir, Interferon, rabbit, Famciclovir, Penciclovir, AZT, DDI, DDC, D4T, abacavir, L-(-)-FMAU, L-DDA phosphate prodrug and β-D-dioxolane nucleosides such as β-D-dioxolane base-guanine (DG), β-D-dioxolane base-2,6-diaminopurine (DAPD), and β-D-dioxolane base-6-chloropurine (ACP), non-nucleoside rt inhibitor such as nevirapine, MKC-442, DMP-266 (sustiva) and proteinase inhibitor such as indinavir, Saquinavir, AZT, DMP-450 etc.

These compounds can also be used for the treatment of equine infectious anemia virus (EIAV), feline immunodeficiency virus and man like ape immunodeficiency virus (Wang, S., Montelaro, R., Schinazi, R.F., Jagerski, B. and Mellors, J.W.: " activity of the anti-equine infectious anemia virus of nucleosides and non-nucleoside reverse transcriptase inhibitor (NNRTI) (EIAV) ", First National Conference on Human Retro viruse and Related lnfections, Washington, DC, 12-16 day in December, 1993; Sellon D.C., " contagious equine abortion ", the Vet.Cl in.North Am.Equine Pract. U.S., 9:321-336,1993; Philpott, M.S., Ebner; J.P., Hoover, E.A.; " estimate 9-(2-phosphoryl methoxy base ethyl) VITAMIN B4 to the treatment of feline immunodeficiency virus with quantitative polyase chain reaction ", Vet.Immunol.Immunopathol.35:155166,1992).

Also provide fluorine has been introduced new on the non-carbohydrate sugar ring precursor and the method for non-enantiomer selectivity thoroughly.The method comprise chirality, non-carbohydrate sugar ring precursor (4S)-5-(the oxygen base of protection)-penta-4-lactone (it can be prepared by Pidolidone) includes but not limited to the reaction of N-fluoro-(two) benzenesulfonimide with the electrophilic source of fluorine, obtains key intermediate fluoro lactone 6.This fluoro lactone is reduced to the acetic ester that lactol and ethanoyl obtain the end group isomery, then for the synthesis of some new β-L-α-2 '-fluoro nucleosides.Also available D-Glu is as the synthetic corresponding D-enantiomer of initiator.

In another interchangeable embodiment, the glycal of fluoridizing of preparation dehydrogenation, then change 2 into ', 3 '-dideoxy-2 ', 3 '-two dehydrogenations-2 '-fluoro nucleosides or β-L or β-D-R base-2 '-the fluoro nucleosides, as mentioned below.

Give the preparation 2 ', 3 '-dideoxy-2 ', 3 '-two dehydrogenations-2 '-simple method of fluoro nucleosides, the method comprises the 6-chloro-purine of silylation and key intermediate direct polycondensation, this key intermediate is by L-2,3-O-acetone glycerol aldehyde (glyceraldenhyde).

Detailed Description Of The Invention

Compound, the method and composition of the HIV that the present invention relates to a kind of people for the treatment of or other animal host disclosed herein, hepatitis (B-mode or the third type) or abnormal cell proliferation; comprising using selectivity to be present in 2 '-fluoro-nucleosides of the significant quantity in the pharmaceutical carrier; a kind of medicinal derivative (be included in 5 '-position or on purine or pyrimidine, carried out the compound of alkylation or acylations), or its pharmaceutical salts.Activity that compound of the present invention has antiviral (being anti-HIV-1, anti-HIV-2 or anti-hepatitis (B-mode or the third type)), or antiproliferative activity, or metabolism is to have these active compounds.

Put it briefly, the present invention includes following feature:

(a) β-L and β-D-2 '-fluoro nucleosides is as described herein, and medicinal derivative and pharmaceutical salts,

(b) β-L and β-D-2 '-fluoro nucleosides is as described herein, and medicinal derivative and pharmaceutical salts, and they are used for pharmacological agent, and for example, treatment or prevention HIV or hepatitis (B-mode or the third type) infect or the treatment abnormal cell proliferation;

(c) 2 ', 3 '-dideoxy-2 ', 3 '-two dehydrogenations-2 '-fluoro-L-glyceryl-penta-2-thiazolinyl-furyl glycosyl nucleosides, and medicinal derivative and pharmaceutical salts, they are used for pharmacological agent, for example, treatment or prevention HIV or hepatitis (B-mode or the third type) infect or the treatment abnormal cell proliferation;

(d) these 2 '-fluoro nucleosides and medicinal derivative thereof and its salt purposes in the medicine of preparation treatment HIV or virus infection or treatment abnormal cell proliferation;

(e) contain the pharmaceutical preparation of 2 '-fluoro nucleosides or its medicinal derivative or its salt and pharmaceutical carrier or thinner;

(f) prepare the method for β-L and β-D-2 '-alpha-fluoro nucleosides, see for details hereinafter, and

(g) preparation 2 ', 3 '-dideoxy-2 ', the method for 3 '-two dehydrogenations-2 '-fluoro-L-glyceryl-penta-2-thiazolinyl-furyl glycosyl nucleosides.

I. active compound, and physiology acceptable derivates and salt

2 ' following-α of structure-fluoro-nucleosides is provided:

R wherein ¹OH, H, OR ³, N ₃, CN, halogen (comprising F) or CF ₃, low alkyl group, amino, low-grade alkyl amino, two elementary alkyl amido or alkoxyl group, and base refers to purine or pyrimidine bases;

R ²Be H, phosphate radical comprises monophosphate root, gen-diphosphate, triphosphate, or stable phosphate prodrug; Acyl group, or other medicinal leavings group, this group can provide R when using in the body ²It is the compound of H or phosphate radical; Sulphonate comprises alkyl or aralkyl alkylsulfonyl (comprising methyl sulphonyl), benzyl (wherein the phenyl selectivity is replaced by the substituting group described in one or more above-mentioned aryl definition), lipid (comprising phosphatide), amino acid, peptide or cholesterol; And

R ³Be acyl group, alkyl, phosphate radical or other medicinal leavings group, namely when using in the body, can be cracked into the group of parent compound.

In second embodiment, provide 2 '-fluoro nucleosides of following formula:

In the 3rd embodiment, provide 2 '-fluoro nucleosides of following formula:

In the 4th embodiment, provide structure following 2 '-fluoro nucleosides:

In this article, unless stated otherwise, the term alkyl refers to saturated straight chain, side chain or ring-type primary, the second month in a season or uncle C ₁To C ₁₀Hydrocarbon, and particularly including methyl, ethyl, propyl group, sec.-propyl, cyclopropyl, butyl, isobutyl-, the tertiary butyl, amyl group, cyclopentyl, isopentyl, neo-pentyl, hexyl, isohexyl, cyclohexyl, cyclohexyl methyl, 3-methyl amyl-2,2-dimethylbutyl, and 2,3-dimethylbutyl.This alkyl can be selected from following part selectivity and replace by one or more: hydroxyl, amino, alkylamino; arylamino, alkoxyl group, aryloxy; nitro, cyano group, sulfonic acid; sulfate radical, phosphonic acids, phosphate radical or phosphonate radical; they can be unprotected or as required protection; as is known to persons skilled in the art, such as Greene etc. Protective Groups inOrganic Synthesis, John Wiley and Sons, second edition, 1991 instruct, and are hereby incorporated by.

Term lower alkyl unless stated otherwise, refers to C in this article ₁To C ₄Saturated straight chain, side chain or ring (if suitably, for example cyclopropyl) alkyl.

Term alkylamino or arylamino refer to respectively with the substituent amino of one or two alkyl or aryl.

Term " protection " refers to be added on Sauerstoffatom, nitrogen-atoms or the phosphorus atom to prevent further reaction or the group that adds for other purpose unless limit separately in this article.The kinds of protect base of Sauerstoffatom or nitrogen-atoms is that the organic synthesis those skilled in the art are known.Herein, term aryl refers to phenyl, xenyl or naphthyl unless stated otherwise, and preferred phenyl.Aryl can selectedly replace from one or more following part selectivity: hydroxyl, amino, alkylamino; arylamino, alkoxyl group, aryloxy; nitro, cyano group, sulfonic acid; sulfate radical, phosphonic acids, phosphate radical or phosphonate radical; they can be unprotected or as required protection; as is known to persons skilled in the art, such as Greene etc. Protective Groups in Organic Synthesis, John Wiley and Sons, second edition, 1991 instruct.

Term alkaryl or alkylaryl refer to the alkyl with aryl substituent.Term aralkyl or arylalkyl refer to the aryl with alkyl substituent.

Term halogen in this article, comprises chlorine, bromine, iodine and fluorine.

Term purine or pyrimidine bases include but not limited to VITAMIN B4, N ⁶-alkyl purine, N ⁶(wherein acyl group is C (O) (alkyl, aryl, alkylaryl, or arylalkyl) to-acyl group purine, N ⁶-benzyl purine, N ⁶-halo purine, N ⁶-vinyl purine, N ⁶-ethyl purine, N ⁶-acyl group purine, N ⁶-hydroxyalkyl purine, N ⁶-alkylthio purine, N ²-alkyl purine, N ²-alkyl-6-thio-purine, thymus pyrimidine, cytosine(Cyt), 5-flurocytosine, 5-methylcytosine, 6-aza-pyrimidine (comprising 6-azepine cytosine(Cyt)), 2-and/or 4-mercaptopyrimidine, uridylic, 5-halo uridylic (comprising 5 FU 5 fluorouracil),

The alkyl pyrimidine, C ₅-benzyl pyrimidines, C ₅-halogenated pyrimidine, 5-vinyl pyrimidine, C ₅-ethynyl pyrimidine, C ₅-acyl group pyrimidine, C ₅-hydroxyalkyl purine, C ₅-amido pyrimidine, C ₅-cyanopyrimidine, C ₅-nitro-pyrimidine, C ₅-aminopyrimidine, N ²-alkyl purine, N ²-alkyl-6-thio-purine, 5-azepine cytosine(Cyt) base, 5-azauracil base, three azepine cyclopentano pyridyl, imidazopyridyl, pyrrolo-pyrimidine radicals and pyrazolopyrimidine bases.Purine bases include but not limited to guanine, VITAMIN B4, xanthoglobulin, 2,6-diaminopurine and 6-chloropurine.Sense oxygen on the base or nitrogen-atoms group can be as required or are claimed.Suitable protecting group is well known to those skilled in the art, and comprises trimethyl silyl, dimethyl hexyl silyl, t-butyldimethylsilyl and tertiary butyl xenyl silyl, trityl, alkyl, acyl group such as ethanoyl and propionyl, methylsulfonyl and p-toluenesulfonyl.

When the patient is carried out administration, can use active compound with the form that parent compound or the activated any derivative of itself tool can directly or indirectly be provided.Limiting examples is pharmaceutical salts (perhaps being called " physiologically acceptable salt "), or 5 '-carried out the compound (perhaps being called " medicinal derivative ") of alkylation or acylations on position or purine or the pyrimidine bases.In addition, modification can affect the biological activity of this compound, increases in some cases the activity of parent compound.By preparing derivative and detecting its antiviral activity according to the method for describing or other method well known by persons skilled in the art herein and can easily estimate this compound.

The term acyl group refers to carboxylicesters, and wherein the non-carbonyl moiety of ester group is selected from straight chain, side chain or cycloalkyl or low alkyl group, and alkoxyalkyl comprises methoxymethyl, and aralkyl comprises benzyl, and aryloxy alkyl such as phenoxymethyl, aryl comprise optionally by halogen, C ₁To C ₄Alkyl or C ₁To C ₄The phenyl that alkoxyl group replaces; sulphonate such as alkyl or aralkyl alkylsulfonyl comprise methylsulfonyl, and be single, two or triguaiacyl phosphate, trityl or mono methoxy trityl; the benzyl that replaces, trialkylsilkl (for example dimethyl tertiary butyl silyl) or diphenyl methyl silyl.Aryl preferably includes phenyl in ester.

In this article, term " there is no " or " basically lack " refers to that nucleotide composition contains at least 95% to 98%, or the more preferably enantiomer of the appointment of this nucleosides of 99% to 100%.

The nucleoside prodrugs preparation

Any nucleosides of describing herein can be with the form administration of nucleoside prodrugs with the activity, bioavailability, the stability that increase this nucleosides or change its character.Some Nucleotide prodrug parts are known.Generally speaking, to nucleosides single, two or triphosphate carry out the stability that alkylation, acylations or other lipophilic modification can increase this Nucleotide.The substituent example of the one or more hydrogen atoms on the replaceable phosphate radical is alkyl, aryl, steroid class, carbohydrate, comprises sugar, 1,2-DG and alcohol.Much be described in R.Jones and N.Bischofberger, AntiviralResearch, 27 (1995) 1-17.Wherein any compound can be united with nucleosides disclosed herein and used to reach required effect.

This active nucleosides can also provide with the form of 5 '-phosphorus ether fat or 5 '-ether fat, as disclosed in the following document, they are incorporated herein by reference: Kucera, L.S., N.lyer, E.Leake, A.Raben, Modest E.K., D.L.W., and C.Piantadosi.1990, " suppressing the new membrane interaction ether lipid that infectivity HIV-1 produces and induced defect virus forms ", AIDS Res.Hum.Retro Viruses.6:491-501; Piantadosi, C., J.MarascoC.J., S.L.Morris-Natschke, K.L.Meyer, F.Gumus, J.R.Suries, K.S.Ishaq, L.S.Kucera, N.Iyer, C.A.Wallen, S.Piantadosi, and E.J.Modest.1991, " new ether fat nucleosides conjugate synthetic also estimated its HIV (human immunodeficiency virus)-resistant activity ", J.Med Chem.34:1408,1414; Hosteller, K.Y., D.D.Richman, D.A.Carson, L.M.Stuhmiller, G.M.T.van Wijk, and H.van den Bosch.1992, " with 3 '-deoxythymidine diphosphate two mnyristoyl base glycerols, a kind of 3 '-the lipid prodrug of deoxythymidine; greatly improved the inhibition to 1 type human immunodeficiency virus replication in CEM and the HT4-6C cell ", Antimicrob.Agents Chemother.36:2025.2029; Hosetler, K.Y., L.M.Stuhmiller, H.B.Lenting, H.vanden Bosch, and D.D.Richman, 1990, " the synthetic and antiretroviral of the phospholipid analogues of azido-thymidine and other anti-viral nucleoside is active ", J.BioL Chem.265:61127.

Disclose and to be connected on the nucleosides by covalent linkage, preferably in the limiting examples of the United States Patent (USP) of the suitable lipophilic substituting group of 5 ' of nucleosides-OH position or lipophilic preparation, comprised United States Patent(USP) Nos. 5,149,794 (on September 22nd, 1992, Yatvin etc.); 5,194,654 (on March 16th, 1993, Hostetler etc.), 5,223,263 (on June 29th, 1993, Hostetler etc.); 5,256,641 (on October 26th, 1993, Yatvin etc.); 5,411,947 (May 2 nineteen ninety-five, Hostetler etc.); 5,463,092 (October 31 nineteen ninety-five, Hostetler etc.); 5,543,389 (on August 6th, 1996, Yatvin etc.); 5,543,390 (on August 6th, 1996, Yatvin etc.); 5,543,391 (on August 6th, 1996, Yatvin etc.); And 5,554,728 (on September 10th, 1996; Basava etc.), all the elements are incorporated herein by reference.The lipophilic substituting group that can connect nucleosides of the present invention or the foreign patent of lipophilic preparation are disclosed, comprise WO 89/02733, WO 90/00555, and WO 91/16920, and WO 91/18914, WO 93/00910, WO 94/26273, and WO 96/15132, and EP 0350287, EP 93917054.4, and WO 91/19721.

The limiting examples of Nucleotide prodrug is described in following document: Ho, D.H.W. (1973) " kinases of 1 β-D-furyl glycosyl cytosine(Cyt) and the distribution of deaminase in people and mouse tissue ", Cancer Res.33,2816-2820; Holy, the nucleotide analog that the utmost point phosphorus such as A. (1993) are modified ", see: De Clercq (editor), Advances In Antiviral Drug DesignVol.1, JAI Press, pp.179-231; Hong, C.I., Nechaev, A., and West, C.R. (1979a) " the synthetic and anti-tumor activity of the 1-β of hydrocortisone and cortisone-D-furyl glycosyl cytosine(Cyt) conjugates ", Bicohem.Biophys.Rs.Commun.88,1223-1229; Hong, C.I., Nechaev, A., Kirisits, A.J.Buchheit, D.J. and West, C.R. (1980) " as the nucleosides conjugates of potential antineoplastic agent, 3. synthetic the and anti-tumor activity of the 1-of reflunomide and selected lipophilic alcohol (β-D-furyl glycosyl) cytosine(Cyt) conjugates ", J.Med.Chem.28,171-177; Hosteller, K.Y., Stuhmiller, L.M., Lenting, H.B.M.van den Bosch, H. and Richman J.Biol.Chem.265,6112-6117; Hosteller, K.Y., Carson, D.A. and Richman, D.D. (1991); " phosphatidyl azido-thymidine: antiviral mechanism in cem cell ", J.Biol.Chem.266,11714-11717; Hosteller, K.Y., Korba, B.Sridhar, C., Gardener, M. (1994a) " antiviral activity of phosphatidyl-zalcitabine in the cell of hepatitis B infection and the liver absorption that in mouse, improves ", Antiviral Res.24,59-67; Hosteller, K.Y., Richman, D.D., Sridhar.C.N.Felgner, P.L.Felgner, J., Ricci, J., Gardener, M.F.Selleseth, D.W. and Ellis, M.N. (1994b) " phosphatidyl azido-thymidine and phosphatidyl-ddC: the evaluation of antiviral activity in the absorption of mouse lymph tissue and the cell the human immunodeficiency virus infection and in rauscher leukemia infecting mouse ", Antimicrobial Agents Chemother.38,2792-2797; Hunston, R.N., Jones, A.A.McGuigan, C., Walker, R.T., Balzarini, J., and DeClercq, E. (1984) " derive from 2 '-some cyclic phosphoric acid three ester synthesis and biological properties of '-Deoxy-5-fluorouridine ", J.Med Chem.27,440-444; Ji, Y.H., Moog, C., Schmitt, G., Bischoff, P. and Luu, B. (1990); " as the 7-beta-hydroxy cholesterol of potential antineoplastic agent and the phosplate of pyrimidine nucleoside: the preliminary assessment of synthetic and anti-tumor activity ", J.Med Chem.33 2264-2270; Jones, A.S., McGuigan, C., Walker, R.T., Balzarini, J. and DeClercq, E. (1984) " synthetic, the character of some nucleosides endoxan and biological activity ", J.Chem.Soc.Perkin Trans.I, 1471-1474; Juodka, B.A. and Smrt, J. (1974) " two ribonucleoside phosphorus (P → N) amino acid derivative is synthetic ", Coll.Czech.Chem.Comm.39,363-968; Kataoka, S., Imai, J., Yamaji, N., Kato, M., Saito, M., Kawada, T. and Imai, S. (1989) " alkylating cAMP derivative; Synthetic and the biological activity of selectivity ", NucleicAcids Res, Sym.Ser.21,1-2; Kataoka, S., Uchida, " (cAMP) benzyl and methyl three esters ", Heterocycles 32,1351-1356; Kinchington, D., Harvey, J.J., O ' Comor, T.J., Jones, B.C.N.M., Devine, K.G., Taylor-RobinsonD., Jeffiies, D.J. and McGuigan, C. (1992) " comparison of the antivirus action of the phosphoramidate of zidovudine and the anti-HIV of phosphorodiamidite derivatives and ULV ", Antiviral Chem.Chemother.3,107-112; Kodmna, K., Morozumi, M., Saithoh, K.I., Kuninaka, H., Yosino, H. and Saneyoshi, M. (1989) " anti-tumor activity and the pharmacology of 1-β-D-furyl glycosyl cytosine(Cyt)-5 '-stearyl phosphoric acid ester; The Orally active derivative of a kind of 1-β-D-furyl glycosyl cytosine(Cyt) ", Jpn.J.Cancer Res.80,679-685; Korty, M. and Engels, J. (1979) " adenosine-and guanosine 3 ', 5 ' phosphoric acid and benzyl ester to the effect of guinea-pig ventricular's cardiac muscle ", Naunyn-Schmiedeberg ' s Arch.Pharmacol.310,103-111; Kumar, A., Goe, P.L., Jones, A.S.Walker, R.T.Balzarini, J. and DeClereq, E. (1990) " the synthetic and biological assessments of some ring phosphoramidate nucleoside derivates ", J.Med.Chem, 33,2368-2375; LeBec, C. and Huynh-Dinh, T. (1991) " the phosphotriester derivative of 5-FUD, a kind of arabinose base is synthetic as the cytosine arabinoside of anticancer prodrug ", Tetrahedron Lett.32,6553-6556; Lichtenstein, J., Barner.H.D. and Cohen, S.S. (1960) " derives from the metabolism of the exogenous Nucleotide of Escherichiacoli. ", J.Biol.Chem.235,457-465; Lucthy, J., Von Daeniken, A., Friederich, J.Manthey, B., Zweifel, J., Schlafter, C. and Bem, M.H. (1981) " the synthetic and toxicity of natural cyano group table sulfo-alkane ", Mitt.Geg.Lebensmittelunters.Hyg.72,131-133 (Chem.Abstr.95,127093); McGigan, C.Tollerfield, S.M. and Riley, P.a. (1989) " some tricresyl phosphate ester synthesis and biological assessments of antiviral Ara ", Nucleic Acids Res.17,6065-6075; McGuigan, C., Dev ine, K.G., O ' Connor, T.J., Galpin, S.A., Jeffries, D.J. and Kinchington, D. (1990a) " anti-HIV-1 compounds 3 '-azido--3 '-some new phosphoramidic acid ester derivatives of deoxythymidine (AZT) synthetic and estimate ", Antiviral Chem.Chemother.I 107-113; McGuigan, C., O ' Connor, T.J., Nicholls, S.R.Nickson, C. and Kinchington, D. (1990b) " synthetic and HIV (human immunodeficiency virus)-resistant activity of the Acidic phosphates ester derivative of the replacement that some of AZT and ddCyd are new ", Antiviral Chem.Chemother.1,355-360; McGuigan, C., Nicholls, S.R., O ' Connor, T.J., and Kinchington, D. (1990c) " as 3 of potential anti-AIDS drugs '-some new Acidic phosphates ester derivatives of the nucleosides modified synthetic ", Antiviral Chem.Chemother.1,25-33; McGuigan, C., Devin, K.G., O ' Cornnor, T.J., and Kinehington, D. (1991) " the synthetic and HIV (human immunodeficiency virus)-resistant activity of some haloalkyl phosphoramidic acid ester derivatives of AZT (AZT); The strong activity of three chloroethyl methoxy propyl aminoacyl compounds ", Antiviral Res.15,255-263; McGuigan, C., Pathirana, R.N., B, J. and DeCiercq, E. (1993 b) " intracellular transport of the biological activity AZT Nucleotide that the aryl phosphoric acids ester derivative by AZT carries out ", J.Med Chem.36,1048-1052.

The alkyl hydrogen phosphate derivative of anti-HIV reagent A ZT may be low than parent ucleosides toxicity.AntiViral Chem.Chemother.5,271-277; Meyer, R.B., Jr., Shuman, D.A. and Robins, R.K. (1973) " purine nucleoside 3 ', 5 '-ring cyano group phosphate synthesis ", Tetrahedron Lett.269-272; Nagyvary, J.Gohil, R.N., Kirchner, C.R. and Stevens, J.D. (1973) " to the research of ring AMP neutral ester ", Biochem.Biophys.Res.Commun.55,1072-1077; Namane, A.Gouyette, C., Fillion, M.P., fillion, G. and HuynhDinh, T. (1992) " improves the brain transhipment of AZT " with glucosyl phosphotriester prodrug, J.Med Chem.35,3039-3044; Nargeot, J.Nerbome, J.M.Engel s, J. and Leser, H.A. (1983) Natl.Acad.Sci.U.S.A.80,2395-2399; Nelson, K.A., Bentrude, W.G.Stser, W.N. and Hutchinson, J.P. (1987) " nucleosides ring 3 ', the position distortion equilibrium problem of the phosphoric acid ring of 5 ' monophosphate.Thymidine benzyl ring 3 ', 5 '-diastereomer of monophosphate ¹H NMR and X-ray crystallography research ", J.Am.Chem.Soc.109,4058-4064; Nerbonne.J.M., Richard, S., Nargeot, J. and Lester, H.A. (1984) " new photoactivation cyclic nucleotide produces in the cell and jumps over aspect ring AMP and cyclo GMP concentration ", Nature 301,74-76; Neumann, J.M., Herve, M., Debouzy, J.C., Guerra, F.I., Gouyette, C., Dupraz, B. and Huyny-Dinh, T. (1989) " synthetic the reaching with NMR of the glucosyl phosphatide of thymidine carried out the research that its cross-film is transported ", J.Am.Chem.Soc.111,4270-4277; Ohno, R., Tatsumi, N., Hirano, M.; Imai, K.Mizoguchi, H., Nakamura, T., Kosaka; M., Takatuski, K., Yajnaya, T., Toyarna K.; Yoshida, T., Masaoka, T., Hashimoto; S., Ohshima, T., Kimura, I.; Ywnada, K. and Kimura, J. (1991) " oral 1-β-D-R base uranyl base cytosine(Cyt)-5 '-stearyl phosphoric acid treatment myelodysplastic syndrome ", Oncology 48,451-455.Palomino, E., Kessle, D. and Horwitz, J.P. (1989) " is used for 2 ', 3 ' di-deoxynucleoside to the dihydropyridine carrier system of the lasting transhipment of brain ", J.Med Chem.32,22-625; Perkins, R.M., Barney, S.Wittrock, R., Clark, P.H., Levin, R.Lambert, D.M., Peneway, S.R., Serafinowska, H.T., Bailey, S.M., Jackson, S., Hamden, M.R.Ashton, R., Sutton, D., Harvey, J.J. and Brown, A.G. (1993) " " activity that the rauscher murine leukemia virus of BRL47923 and the anti-mouse of oral prodrugs SB203657A thereof infects ", Antiviral Res.20 (Suppl.I) .84; Piantadosi, C., Marasco, C.J., Jr., Norri s-Natschke, S.L., Meyer, K.L., Gumus, F., Surles, J.R., Ishaq, K.S., Kucera, L.S.lyer, N., Wallen, C.A., Piantadosi, S. and Modest, E.J. (1991) " evaluation of the synthetic and Anti-HIV-1 Active of new ether fat nucleosides conjugates ", J.Med Chem.34,1408-1414; Pompon, A., Lefebvre, I., Imbach, J.L., Kahn, S. and Farquhar, D. (1994). " azido-thymidine-5 in cell extract and tissue culture medium (TCM) '-list of monophosphate and the decomposition path-ways of two (oxy acid methyl neopentyl) esters; The application of closed circuit ISRP-cleaning HPLC technology ", Antiviral Chem Chemother.5,91-98; Postemark, T. (1974) " ring AMP and cyclo GMP ", Annu.Rev.Pharmacol.14,23-33; Prisbe, E.J., Martin, J.C.M., McGhee, D.P.C., Barker, M.F., Smee, D.F.Duke, A.E., Matthews, T.R. and Verheyden, J.P.J. (1986) " 9-[(1,3-dihydroxyl-2-propoxy-) methyl] phosphoric acid of guanine and the synthetic and herpesvirus resisting activity of phosphonate derivative ", J.Med.Chem.29,671-675; Pucch, F., Gosselin, G., Lefebvre, I., Pompon, a., Aubertin, A.M.Dim, and Imbach, J.L. (1993) " activation method by the reductase enzyme mediation carries out the intracellular transport of nucleosides monophosphate ", Antivral Res.22,155-174; Pugaeva, V.P..Klochkeva, S.I., Mashbits, F.D. and Eizengart, R.S. (1969). " toxicity assessment of thiirane and health standards grade in industrial environment ", Gig.Trf.Prof.Zabol.14,4748 (Chem.Abstr.72,212); Robins, R.K. (1984) " nucleotide analog is as the potentiality of retrovirus and tumor inhibitor ", Pharm.Res.11-18; Rosowsky, A., Kim.S.H., Ross and J.Wick, M.M. (1982) " lipotropy 5 ' (alkylphosphonic acid carboxylic acid) ester of 1-β-D-furyl glycosyl cytosine(Cyt) and N4-acyl group and 2.2 ' thereof-dehydration-3 ' O-acyl derivative is as potential prodrug ", J.Med Chem.25,171-178; Ross, W. (1961) " progressively loading device is to the increase with aryl nitrogen mustard susceptibility after glucose pretreatment of basic side chain ", Biochem.Pharm.8,235-240; Ryu, E.K., Ross, R.J.Matsushita, T., MacCoss, M., Hong, C.I. and West, C.R. (1982). " phosphatide nucleosides conjugates.3. to 1-β-D-furyl glycosyl cytosine(Cyt) 5 ' bisphosphate [-], the synthetic and preliminary biological assessment of 2-DG ", J.Med Chem.25,13221329; Saffhill, R. and Huine, W.J. (1986) " serum of different sources to the degraded of idoxuridine and 5-bromine oxethyl uridine and the result who mixes DNA with these compounds ", Chem.Biol.Interact.57,347-355; Saneyoshi, M., Morozumi, M., Kodama, K., Machida, J., Kuninaka, A. and Yoshino, H. (1980) " synthetic nucleosides and Nucleotide; a series of 1-β of XVI.-D-furyl glycosyl cytosine(Cyt) 5 '-alkyl or aryl phosphate synthesis and biological assessment ", Chem Pharm.Bull.28,2915-2923; Sastry, J.K., Nehete, P.N., Khan, S., Nowak, B.J., Plunkett, W., Arlinghaus, R.B. and Farquhar, D. (1992) " infection of film perviousness di-deoxyuridine 5 '-monophosphate analogue HIV inhibiting ", Mol.Pharmacol.41,441-445; Shaw, J.P., Jones, R.J.Arimilli, M.N., Louie, M.S., Lee, W.A. and Cundy, K.C. (1994) " derives from the oral administration biaavailability of PMEA in male Sprague-Dawley rat of PMEA prodrug ", 9th Annual AIPS Meeting, San Diego, CA (digest).Shuto, S., Ueda, S., Imamura, S., Fukukawa, K.Matsuda, A. and Ueda, T. (1987) " uses easily one-step synthesis 5 ' phosphatidyl nucleosides of enzyme two phase reaction ", Tetrahedron Lett.28,199-202; Shuto, S.Itoh, H., Ueda, S., Imamura, S., Kukukawa, K., Tsujino, M., Matsuda, A. and Ueda, T. (1988) Pharm.Bull.36,209-217.The example of favourable phosphoric acid ester prodrug class is S-acyl group-2-thio-ethyl class, is also referred to as " SATE ".

II. unite and alternating treatment

Have recognized that and treating the anti-medicine variant that HIV and HBV can occur with antiviral agent for a long time.Resistance is the most normal to be used for the enzyme of virus replication by coding the sudden change of gene produces, and in the situation of HIV, the most generally reversed transcriptive enzyme, proteolytic enzyme or archaeal dna polymerase, and for HBV, be archaeal dna polymerase.Recently, shown by causing the second of different sudden changes from main ingredient, perhaps the third antiviral compound associating or be used alternatingly this compound, the effect that the anti-HIV of medicine infects can prolong, increases or keep.Perhaps, by this associating or alternating treatment, the pharmacokinetics of this medicine, bio distribution or other parameter can change.In a word, combination therapy generally than alternating treatment more by preferably, this is because it causes multiple stimulation stress to virus.

In one embodiment, the second antiviral agent that treatment HIV uses can be reverse transcriptase inhibitors (a kind of " RTI "), and it can be nucleosides (a kind of " NRTI ") or the non-nucleoside compound (a kind of " NNRTI ") that synthesizes.In one embodiment, for HIV, the second (or the third) antiviral agent can be proteinase inhibitor.In another embodiment, the second (or the third) can be the pyrophosphate analogue or merge binding inhibitors.Some antiviral compounds resistance data external and that body is interior are seen Schinazi etc., the reverse transcription virus gene sudden change relevant with resistance, International Antiviral News, 1997.

The preferred compound that is used for associating or alternating treatment HBV comprises 3TC, FTC, L-FMAU, Interferon, rabbit, β-D-dioxolane base-guanine (DXG), β-D-dioxolane base 2,6-diaminopurine (DAPD) and β-D-dioxolane base-6-chloropurine (ACP), Famciclovir, Penciclovir, BMS-200475, bis pom PMEA (adefovir, dipivoxil); Lobucavir, ganciclovir and ribavirin.

Can comprise suitable-2-methylol-5-(5-flurocytosine-1-yl)-1,3-oxathiolane (FTC) with the preferred embodiment of the antiviral agent of compound disclosed herein associating or alternating treatment; 2-methylol-5-(cytosine(Cyt)-1-yl)-1, (-)-enantiomer of 3-oxa-thiacyclopentane (3TC); Carbovir, acyclovir, FOSCARNET, Interferon, rabbit, AZT, DDI, DDC, D4T, CS-87 (3 '-azido--2 ', 3 '-dideoxy-uridine) and β-D-dioxolane nucleosides such as β-D-dioxolane base-guanine (DXG), β-D-dioxolane base-2,6-diaminopurine (DAPD) and β-D-dioxolane base-6-chloropurine (ACP), MKC-442 (6-benzyl-1-(ethoxyl methyl)-5-sec.-propyl uridylic.

Preferred proteinase inhibitor comprises the full (crixivan of gram filter, Merck), viracept see nelfinaivr (nelfinavir, Agouron), ritonavir (ritonavir, Abbott), Saquinavir (Roche), DMP-266 (Sustiva, efavirenz) and DMP-450 (DuPont Merck).

The more comprehensive tabulation that can unite or replace the compound of administration with any disclosed nucleosides comprises (1S, 4R)-4-[2-amino-6-cyclopropylamino)-9H-purine-9-yl]-2-cyclopentenes-1-methyl alcohol succinate (" 1592 ", a kind of Carbovir analogue; GlaxoWellcome); 3TC:(-)-B-L-2 ', 3 '-dideoxy-3 '-thia cytidine (GlaxoWellcome); A-APA R18893:a-nitro-phenylamino-phenyl-acetamides; A-77003; The proteinase inhibitor of C2 symmetry (Abbott); The symmetrical proteinase inhibitor (Abbott) of A-75925:C2; AAP-BHAP: two heteroaryl piperazine analogues (Upjohn); The symmetrical proteinase inhibitor (Abbott) of ABT-538:C2; AzddU:3 '-azido--2 ', 3 '-di-deoxyuridine; AZT:3 '-azido--3 '-deoxythymidine (GlaxoWellcome); AZT-p-ddI:3 '-azido--3 '-deoxythymidine base-(5 ', 5 ')-2 ', 3 '-dideoxy inosinic acid (Ivax); BHAP: two heteroaryl piperazines; BILA 1906:N-{1S-[[[3-[2S-{ (1,1-dimethyl ethyl) amino] carbonyl }-4R-] the 3-pyridylmethyl) sulfo-]-piperidino]-2R-hydroxyl-1S (phenyl methyl) propyl group] amino] carbonyl]-the 2-methyl-propyl }-2-quinoline formyl amine (Bio Mega/Boehringer-Ingelheim); BILA 2185:N-(1, the 1-dimethyl ethyl)-and 1-[2S-[[2-2, the 6-dimethyl phenoxy)-the 1-oxoethyl) amino]-2R-hydroxy-4-phenyl butyl]-the 4R-pyridylthio)-2-piperidyl urea (BioMega/Boehringer-Ingelheim); BM+51.0836: thiazole also-the isoindoline ketone derivatives; BMS 186,318: aminodiol derivatives HIV-1 proteinase inhibitor (Bristol-Myers-Squibb); D4API:9-[2,5-dihydro-5-(phosphoryl methoxy base)-2-furyl] VITAMIN B4 (Gilead); D4C:2 ', 3 '-two dehydrogenations-Zalcitabine; D4T:2 ', 3 '-two dehydrogenations-3 '-deoxythymidine (Bristol-Myers-Squibb); DdC; Zalcitabine (Roche); DdI:2 ', 3 '-didanosine (Bristol-Myers-Squibb); DMP-266:1,4-dihydro-2H-3,1-benzo _ piperazine-2-ketone; DMP-450:{[4R-(4-a, 5-a, 6-b, 7-b)]-six hydrogen-5, two (hydroxyl)-1 of 6-, 3-two (3-is amino) phenyl] methyl)-4, two (the phenyl methyl)-2H-1 of 7-, 3-diazacyclo heptantriene-2-ketone }-double A sulphonate (Avid); DXG:(-)-β-D-dioxolane guanosine (Triangle); EBU-dM:5-ethyl-1-ethoxyl methyl-6-(3,5-dimethyl benzyl) uridylic; E-EBU:5-ethyl-1-ethoxyl methyl-6-benzyluracils; DS: T 500; E-EPSeU:1-(ethoxyl methyl)-(6-phenyl selenyl)-5-ethyl uracil; E-EPU:1-(ethoxyl methyl)-(6-thiophenyl)-5-ethyl uracil; FTC: β-2 ', 3 '-dideoxy-5-fluoro-3 '-thia cytidine (Triangle); HBY097:S-4-isopropoxy carbonyl-6-methoxyl group-3-(methylthio group-methyl)-3,4-dihydro-quinoxaline-2 (1H)-thioketones; The HEPT:1-[(2-hydroxyl-oxethyl) methyl]-6-(thiophenyl) thymus pyrimidine; HIV-1: human immunodeficiency virus type 1; JM2763:1,1 '-(trimethylene)-two-Isosorbide-5-Nitrae, 8,11-tetraazacyclododecane tetradecane (JohnsonMatthey); JM3100:1,1 '-[Isosorbide-5-Nitrae-phenylene two-(methylene radical)] is two-Isosorbide-5-Nitrae, 8,11-tetraazacyclododecane tetradecane (Johnson Matthey); KNI-272: contain (2S, 3S)-3-amino-tripeptides of 2-hydroxy-4-phenyl butyric acid; L-697,593; 5-ethyl-6-methyl-3-(2-phthalimido-ethyl) pyridine-2 (1H)-ketone; L-735,524: hydroxyl-aminopentane methane amide HIV-1 proteinase inhibitor (Merck); L-697,661:3-{[(-4,7-two chloro-1,3-benzo _ azoles-2-yl) methyl] amino }-5-ethyl-6-picoline-2 (1H)-ketone; L-FDDC:(-)-β-L-5-fluoro-Zalcitabine; L-FDOC:(-)-β-L-5-fluoro-dioxolane cytosine(Cyt); NMC442:6-benzyl-1-ethoxyl methyl-5-sec.-propyl uridylic (I-EBU; Triangle/Mitsubishi); Nevirapine: 11-cyclopropyl-5,11-dihydro-4-methyl-6H-two pyrido 1[3,2-b:2 ', 3 '-e] diazacyclo heptantriene-6-ketone (Boehringer-Ingelheim); NSC648400:1-benzyloxymethyl-5-ethyl-6-(α-pyridine thio) uridylic (E-BPTU); P9941:[2-pyridyl ethanoyl-IlePheAla-y (CHOH)] 2 (Dupont Merck); PFA: phosphonoformate (Trisodium phosphonoformate hexahydrate; Astra); PMEA:9-(2-phosphoryl methoxy base ethyl) VITAMIN B4 (Gilead); PMPA:(R)-9-(2-phosphoryl methoxy base propyl group) VITAMIN B4 Gilead); Ro 31-8959: aminoethyle alcohol derivative HIV-1 proteinase inhibitor (Roche); RPI-312: the peptidyl proteinase inhibitor, 1-[(3s)-3-(just-α-carbobenzoxy-(Cbz))-the 1-asparagyl)-amino-2-hydroxy-4-phenyl butyryl radicals]-just-tertiary butyl-1-proline(Pro) acid amides; 2720:6-chloro-3,3-dimethyl 4-(isopropyl alkenyloxycarbonyl)-3,4-dihydro-quinoxaline-2 (1H) thioketones; SC-52151: hydroxyethyl-hydroxyethylurea isostere proteinase inhibitor (Searle); SC-55389A: hydroxyethyl-hydroxyethylurea isostere proteinase inhibitor (Searle); TIBO R82150:(+)-(5S)-4,5,6,7-tetrahydrochysene-5-methyl-6-(3-methyl-2-butene base) imidazo [4,5,1-jk] [Isosorbide-5-Nitrae]-benzodiazepine cycloheptatriene-2 (1H)-thioketones (Janssen); TIBO 82913:(+)-(5S)-4,5,6,7-tetrahydrochysene-9-chloro-5-methyl-6-(3-methyl-2-butene base) imidazo [4,5,1jk]-[Isosorbide-5-Nitrae] benzo-diazacyclo heptantriene-2 (1H)-thioketones (Janssen); TSAO-m3T:[2 ', 5 '-two-O-(t-butyldimethylsilyl)-3 '-spiral shell-5 '-(4 '-amino-1 ', 2 '-oxa-thiol (thiole)-2 ', 2 '-dioxide)]-b-D-furan pentose base-N3-methyl thymus pyrimidine; The U90152:1-[3-[(1-methylethyl)-amino]-the 2-pyridyl]-the 4-[[5-[(methyl sulphonyl)-amino]-1H-indoles-2-yl] carbonyl] piperazine; UC: sulfo-anilid derivative (Uniroyal); UC-781=N-[4-chloro-3-(3-methyl-2-butene oxygen base) phenyl]-2-methyl-3-furans thioformamide; UC-82=N-[4-chloro-3-(3-methyl-2-butene oxygen base) phenyl]-2-methyl-3-thiophene phenol thioformamide; VB 11,328: hydroxyethyl-sulphonamide proteinase inhibitor (Vertex); VX-478: hydroxyethyl sulphonamide proteinase inhibitor (Vertex); XM 323: ring urea proteinase inhibitor (Dupont Merck).

The combination therapy proliferative disorders

In another embodiment, when as antiproliferative agents, these compounds can be united use with the another kind of compound that increases this result for the treatment of, so-called another kind of compound includes but not limited to antifolic, 5-FU (comprising 5 FU 5 fluorouracil), cytidine analog such as β-L-1,3-dioxolane base cytidine or β-L-1,3-dioxolane base 5-fluorine cytidine, antimetabolite (comprises the purine antimetabolite, cytarabine, fudarabine, floxuridine, Ismipur, methotrexate and 6-thioguanine), hydroxyurea, mitotic inhibitor (comprises CPT-11, Etoposide (VP-21), taxol and catharanthus alkaloid such as vincristine(VCR) and vincaleucoblastine, alkylating agent (includes but not limited to busulfan, Chlorambucil, endoxan, ifosfamide, mustargen, melphalan and thiophene are for group), non-classical alkylating agent, the compound of platiniferous, bleomycin, antitumor antibiotics, anthracycline antibiotics such as Zorubicin and dannomycin, amerantrone class, Topoisomerase II inhibitors, hormone preparation (includes but not limited to reflunomide (dexamethasone, prednisone and Methyllprednisolone), male sex hormone such as Fluoxymesterone and methyltestosterone, oestrogenic hormon such as stilboestrol, estrogen antagonist such as tamoxifen, the general amine of p-GLU-HIS-TRP-SER-TYR-D-TRP-LEU-ARG-PRO-GLY-NH2 such as profit, androgen antagonist such as flutamide, aminoglutethimide, Magace and medroxyprogesterone), asparaginase, carmustine, chlorethyl cyclohexyl nitrosourea, hexamethyl-trimeric cyanamide, Dacarbazine, mitotane, streptozocin, cis-platinum, carboplatin, LEVAMISOLE HCL and folinic acid.Compound of the present invention also can be united use with enzyme treatment agent and immune system toner such as Interferon, rabbit, interleukin-, tumour necrosis factor, macrophage colony stimulating factor and G CFS.

III. the method for preparing active compound

In one embodiment of the invention, provide fluorine has been introduced diastereo-isomerism selective reaction on the sugar moieties of novel nucleoside analogs.This synthesizes can be used for preparing purine and pyrimidine derivatives.Committed step in the synthetic route is chirality, non-carbohydrate sugar ring precursor (4S)-5-(protected-the oxygen base)-penta-4-lactone; for example; (4S)-the electrophilic fluorine source of 5-(tert-butyl diphenyl first siloxanes) penta-4-methyl esters 4 uses; include but not limited to N-fluoro-(two) benzenesulfonimide 5.This type of relatively new N-fluorine sulfimide reagent is at first open in 1984 by Barnette, from then on several times makes with extra care Electron Affinities fluorine (Barnette, W.E.J.Am.Chem.Soc.1984,106,452. of also conduct facility and hyperergy; Davis, F.A.; Han; W., Murphy, C.K.J.Org.Chem.1995,60,4730; Snieckus, V.; Beaulieu, F.; Mohri, K.; Han, W.; Murphy, C.K.; Davis, F.A.Tetrahedron Lett.1994,35 (21), 3465).These reagent are the most frequently used to be transitted to nucleophilic reagent such as enolate and metallized aromatics (Davis, F.A. with fluorine; Han; W., Murphy, C.K.J.Org.Chem.1995,60,4730).Specifically, N-fluoro-(two) benzenesulfonimide (NFSI) is stable in the air, easy-to-handle solid, to the stereoselectivity of the enolate of the lactone 4 of silyl protection is fluoridized have enough sterically hindered.As the limiting examples of this method, fluoridize the synthetic of lactone 6 and details are as follows as the common intermediate in more synthetic new α-2 '-fluoro nucleosides.According to this specification sheets, those of ordinary skills as required conventional this method of revising also prepare the compound that needs with the purpose of finishing requirement.

Can be with the Electron Affinities fluorine in any source with precursor (4S)-5-(protected-the oxygen base)-penta-4-lactone, for example (4S)-5-(tert-butyl diphenyl siloxy-)-penta-4-lactone is fluoridized.Other source of Electron Affinities fluorine comprises N-fluoro thionamic acid class (Differding etc., Tet.Lett.Vol.29, No.47pp.6087-6090 (1988); Chemical Reviews, 1992, Vol92, No.4 (517)), N-fluoro-O-benzene disulfonyl imines (Tet.Lett.Vol.35,3456-3468 page or leaf (1994), Tet Lett.Vol35.No.20,3263-3266 page or leaf (1994)); J.Org.Chem.1995,60,4730-4737), 1-vinyl fluoride and synthetic coordinator (Matthews, Tet.Lett.Vol.35, No.7,1027-1030 page or leaf (1994); Allied Signal, Inc., Buffalo Research Laboratory, Buffalo, Accufluor fluorination reagent (NFTh (the 1-fluoro-4-hydroxyl-1 that New York sells, 4-two azo-groups-dicyclo [2.2.2] octane two (a tetrafluoro borate)), NFPy (N-fluorine pyridine _ pyridine seven fluorine biborates) and NFSi (N-fluorobenzene sulfimide); Aldrich Chemical Company, Inc. the Electron Affinities fluorination reagent of selling comprises N-fluorine pyridine _ salt ((1-fluoro-2,4,6-trimethylpyridine _ fluoroform sulphonate, 3,5-, two chloro-1-fluorine pyridine _ fluoroform sulphonates, 1-fluorine pyridine _ fluoroform sulphonate, 1-fluorine pyridine _ a tetrafluoro borate and 1-fluorine pyridine _ seven fluorine biborates), also can be referring to J.Am.Chem.Soc., Vol 112, No.23 1990); N-fluorine sulfimide and amides (N-fluoro-N-methyl-p-toluol sulfonamide, N-fluoro-N-propyl group-p-toluol sulfonamide and N-fluorobenzene sulfimide); The peaceful fourth fluorochemical of N-fluoro-quinoline (J.Chem.Soc.Perkin Trans I 1988,2805-2811); Perfluor-2,3,4,5-tetrahydropyridine and perfluor-(1-crassitude), Banks, Cheng and Haszeldine, Polyfluoro generation-compound part the II of heterocycle(1964); 1-fluoro-2-pyridone, J.Org.Chem., 1,983 48,761-762; Quaternary stereogenic center (T.Chem.Soc.Perkin.Trans.221-227 page or leaf (1992)) with fluorine atom; N-fluoro-2,4,6-pyridine _ fluoroform sulphonate, Shimizu, Tetrahedron Vol 50 (2), 487-495 page or leaf (1994); N-fluorine pyridine _ pyridine seven fluorine biborates, J.Org.Chem.1991,56,5962-5964, Umemoto etc., Bull.Chem.Soc.Jpn., 64 1081-1092 (1991); N-fluoro perfluoroalkyl sulfimide class, J.Am.Chem.Soc., 1987,109,7194-7196; Purrington etc., the aromatic substrate that Lewis is acid mediated fluoridize J.Org.Chem.1991,56,142-145.

The remarkable advantage of this method is can be by suitably selecting respectively " natural " that L-or D-Glu initiator use respectively nucleosides (1a) (1b) L enantiomorph of D or " non-natural ".

By the route shown in the scheme 1 by the synthetic lactone 4 of Pidolidone, such as described (Tetrahedron 1974,30,3547) such as (Tetrahedron 1978,34,1449) as described in the Ravid etc. and Taniguchi.

Scheme 1

The enolate of known lactone 4 prepares in THF with LiHMDS at-78 ℃, is stable.Carried out some synthesizing with this enolate, comprised with high yield adding electrophilic reagent such as diphenyl disenenide compound, Diphenyl disulfide compound and alkyl halide (Liotta, D.C.; Wilson, L.J.Tetrahedron Lett.1990,31 (13), 1815; Chu, C.K.; Babu, J.R.; Beach, J.W.; Ahn.S.K.; Huang, H.; Jeong, L.S.; Lee .S.J.J.Org.Chem., 1990,55,1418; Kawakami, H.; Ebata, T.; Koseki, K.; Matsushita, H.; Naoi, Y.; Ltoh, K.Chem.Lett.1990,1459).But, add the yield extreme difference of required single fluoro product 6 that 5 THF solution obtains in 4 the enolate.Formed some by products and comprised the two fluoro lactones of inferring, itself and other impurity is inseparable.For this reason, change the addition sequence of reagent, be dissolved in together lactone 4 and NFSi5 among the THF and be cooled to-78 ℃.Slowly add LiHMDS, obtain the unique product except unreacted starting material (reaction formula 1) in a small amount.

Reaction formula 1

After silica gel chromatography and crystallization, can obtain the fluorine lactone 6 of yield 50-70%.This reaction obtains 6 single diastereomer, and supposition is because the interaction of the larger fluorination reagent 5 of sterically hindered TBDPS group and volume.Detect by NMR data and x-ray crystal structure more previously disclosed, its enantiomer 20, determine that fluoro lactone 6 is the fluorine isomer of α or " downwards ".

Shown in scheme 2, change lactone 6 into anomer acetic ester 8.Should notice that 7 forms with the β anomer of lactol exist, and acetic ester 8 does not show the detectable α anomer of NMR, such as reports (Bull.Chem.Soc.Jpn.1995,68,1509) such as Niihata.

Scheme 2

By standard Vorbruggen method coupling 8 and silylated pyrimidine bases (Tetrahedron Lett.1978,15,1339), sour as Lewis with the TMS fluoroform sulphonate.Perhaps, can use any other known condensation base and carbohydrate of can being used for to form the Lewis acid of nucleosides, comprise tin chloride, titanium chloride and other tin or titanium compound.Some bases with the column chromatography purifying after the successfully coupling of high yield (reaction formula 2, table 1) of 72-100%.

Reaction formula 2

Table 1. with 8 with the miazines glycosylations that replace

Proton N MR shows that the ratio of β and α nucleosides anomer is about 2: 1 in all cases.The nucleosides of silyl protection can not be split as by column chromatography the anomer of separation.But, use NH ₄F in methyl alcohol with 5 '-the Sauerstoffatom deprotection after (reaction formula 3), can easily α be separated with the β anomer, and be summarized in table 2.

Reaction formula 3

The deprotection of table 2. nucleosides

The classification of the free nucleosides of α or β is based on the chemical shift (table 3) of end group isomery proton and as the polarity of the nucleosides observed by thin-layer chromatography.Observe the right trend of α/β of all free nucleosides, the chemical shift of the anomeric proton of the compound that its Semi-polarity is less obviously than the larger compound of polarity in higher magnetic field.

Table 3. end group isomery chemical shift of proton (ppm)

By comparing 18a (Niihata, S.; Ebata, T.; Kawakami, H.; Matsushida, H.Bull.Chem.Soc.Jpn.1995,68,1509) and 18b (Aerschot, A.V.; Herdewijn, P.; Balzarini, J.; Pauwels, R.; De Clercq, E.J.MedChem.1989,32,1743), measure with former disclosed spectral data and by the X ray crystalline texture of 14b and 15b, prove the relation of end group isomery chemical shift of proton and absolute structure.This discovery is opposite with the common trend of nucleosides, wherein at this in the two the general polarity of α anomer less.Probably " downwards " 2 '-fluoridize in the nucleosides, the C-N end group isomery bond dipoles in the strong dipole antagonism beta isomer of C-F key also reduces the dipolar nature of whole molecule.On the contrary, the geometric configuration of α anomer makes molecular dipole by adding C-F and the C-N bond dipoles is strengthened.Therefore, for the situation of α-2 '-fluoro nucleosides, the α anomer has more polarity than β anomer.

α can not separate by column chromatography with 17b with β anomer 17a because free amino make nucleosides silica gel walk too fast.Therefore, need to use N ⁴The ethanoyl cytosine(Cyt) prepares 11 and also splits 16a and 16b again.Saturated methanol solution with ammonia is quantitatively removed N ⁴-ethanoyl is so that the 17a that obtains separating and 17b.When with 5-flurocytosine during as alkali (compound 10), anomer 15a separates easily with 15b, and does not observe at silica gel and form spot.

In ten kinds of nucleosides that table 2 is listed, as if only have 17b (Martin, J.A.; Bushnell, D.I.; Duncan, I.B.; Dunsdon, S.J.; Hall, M.J.; Machin, P.J.; Merrett, J.H.; Parkes, K.E.B.; Roberts, N.A.; Thomas, G.J.; Galpin, S.A.; Kinchington, D.J.Med.Chem.1990,33 (8), 2137; Zenchoff, G.B.; Sun, R; Okabe, M.J.Org.Chem.1 991,56, and 4392), 18a (Niihata.S.; Ebata, T.; Kawakami, H.; Matsushida, H.Bull.Chem.Soc.Jpn.1995,68,1509) and 18b (Aerschot, A.V.; Herdewijn, P.; Balzarini, J.; Pauwels, R.; De Clercq, E.J.Med Chem.1989,32,1743) in the past synthetic.They as some are known 2 '-β or " making progress " fluorinated nucleoside analogue ¹⁴The same synthetic by natural precursor (being that they are β-D configurations).As if do not determine β-L-2 '-fluoro ribofuranose yl nucleosides in the document before the present invention.

General these molecules are introduced fluorine (Mengel, R. by nucleophillic attack dehydration nucleosides; Guschlbauer, W.Angew.Chem., Int.Ed.Engl.1978,17,525) or by fluoridizing the fixing hydroxyl displacement of diethylamino sulphur (DAST) stereochemistry or be converted introducing fluorine (Herdewijn, P. with three; Aerschot, A.V.; Kerremans, L.NucleosidesNucleotides, 1989,8 (1), 65).One of advantage of the inventive method is not need hydroxyl for introducing fluorine.Therefore, the method is not limited only to natural nucleus glycoside or sugar as initiator, and make 2 '-use of the non-natural enantiomer of fluoro nucleosides becomes simple.

Therefore, some non-natural nucleosides have been synthesized with D-Glu 19 as initiator (scheme 3) with this synthetic route.Sugar ring precursor 20 fluoridize with aforesaid method and from different silylated base coupling (table 4).

Scheme 3

The yield of table 4. non-natural nucleoside analogue

Scheme 4

29 success is synthesized shown in scheme 4, has utilized two class nucleosides.The first kind is to be called 2 ', 3 '-dideoxy-2 ', 3 '-two dehydrogenations-2-2 '-fluoro-nucleosides, and 30, and being nucleosides, Equations of The Second Kind " makes progress "-fluoro or arabinose base analogue, 31, as described in following scheme 5.

Scheme 5

Can synthetic compound 30 and 31 by common intermediate 32, it can obtain by the selenizing of fluoro glycal 29.

Scheme 6

By the compound 32 of selenizing being changed into " making progress " fluoro analogue 31 with the Raney nickel reduction.Perhaps, use NaIO ₄Or hydrogen peroxide is selenide 32 oxidations, and the thermal elimination by this selenide intermediate obtains 30 subsequently.To not fluoridizing these conversions existing play-by-play (Wurster, the J.A. of system; Ph.D.Thesis, Emory University, 1995; Wilson, L.J.; Ph.D.Thesis, Emory University, 1992).

In addition, the synthetic of nucleosides 30 and 31 enantiomer also is possible, and this is because they derive from 29 enantiomer.

Compound 2 ', the 3 '-dideoxy-2 ' of preparation 30 expressions, the square case 7 of the another kind of route of 3 '-two dehydrogenations-2 '-fluoro-nucleosides.This route provide with large-scale silylated base come simply, directly to obtain this compounds method and successfully finish.

Scheme 7

But form silylated ketene acetal Stereoselective by 6 and add the phenyl Selenium monobromide to produce single isomeric compound 36.The reduction of this compound and acylation reaction are carried out smoothly and two step yields height, obtain 37.It is stereoselective that the α of phenyl selenyl orientation makes in subsequently the glycosylation step, and has finished beta isomer synthetic of nucleosides 38 with good yield.Compound 38 can be with the be eliminated product 39 of reaction of hydrogen peroxide oxidation in methylene dichloride, but with our experience, only need to be adsorbed onto on the silica gel and allow it stop some hours 38, after this can be to be close to quantitative yield wash-out from piston boit.Remove as previously mentioned the nucleosides product that 39 protecting group obtains end product 30 and obtains yield good (81%).

Scheme 8

A series of chemical transformations for the synthesis of 30 and 31 can be used for synthetic 34 and 35 equally.

Experimental section

General method

N-fluoro-(two) benzenesulfonimide 5 derives from Allied Signal, not purified direct use.Other all reagent derives from the not purified direct use of Aldrich Chemical Company.Detect fusing point and not calibration with Thomas Hoover capillary melting point apparatus.Obtain the IR spectrum with Nicolet Impact400FT-IR spectrograph. ¹H NMR and ¹C NMR spectrum record on NT-360 or Varian400MHz chromatographic instrument.The TLC plate is silica gel 60F ₂₅₄(0.25mm is thick) is available from EM Science.Flash chromatography is in that (the 230-400 order carries out on ASTM) available from the silica gel 60 of EM Science.All reactions are carried out under the argon atmospher of drying in flame-dried glass reactor.By the rotary evaporation desolventizing.By Atlantic Microlab, Inc, Atlanta, GA. carries out ultimate analysis.

(2S, 4R)-5-(t-butyldiphenylsilyl oxygen)-2-fluoro penta-4-lactone (20)

Add (4R)-5-(tert-butyl diphenyl siloxy-) of being present among the anhydrous THF of 250mL-penta-4-lactone (20.0g in the reaction flask, 0.0564mol, 1.0eq.) and N-fluoro-(two) benzenesulfonimide (NFSi) 5 (17.80g, 0.0564mol, 1.0eq.).This solution is cooled to-78 ℃, and in 1 hour, drips the 1.0MTHF solution of 68.0mL (0.0680mol, 1.2eq.) LiHMDS.Again it was stirred 2 hours under-78 ℃, then be warming up to room temperature and restir 1 hour.After finishing reaction, stop this reaction with the 10mL saturated ammonium chloride solution.Dilute this mixture and pour in isopyknic saturated sodium bicarbonate with the ether of 3 volumes.Again wash organic layer and with saturated sodium-chloride washing once with saturated sodium bicarbonate.With this organic layer of dried over mgso, filter and the concentrated yellow oil that obtains.With this oily matter by silica gel column chromatography with 30% ether/70% hexane solvent system purifying.Again with the gained white solid with hot hexane crystallization, obtain the transparent crystalline solid of 13.04g (yield 62%): R _f(30% ether/70% hexane)=0.26; Mp115-116 ℃.

1H NMR (360MHz, CDCl ₃) d7.63-7.60 (m, 4H), 7.45-7.35 (m, 6H), (5.49 dt, J=52.9 and 7.9Hz, 1H), 4.69 (d, J=9.36Hz, 1H), 3.91 (d, J=11.5Hz, 1H), 3.60 (d, J=11.5Hz, 1H), 2.72-2.40 (m, 2H), 1.05 (s, 9H); ¹³C NMR (100MHz, CDCl ₃) d172.1 (d, J=20.5Hz), 135.5,135.4,132.3,131.7,130.1,128.0,127.9,85.6 (d, J=186.6Hz), 77.3 (d, J=5.3Hz), 65.0,31.8 (d, J=20.5Hz), 26.7,19.1; IR (film) 2958,1796,1252,1192,1111,1016cm ^-1HRMS[M+Li] C ₂₁H ₂₅O ₃FSiLi theoretical value: 379.1717.Measured value: 379.1713.CHAFFS ultimate analysis theoretical value: C, 67.71; H, 6.76.Measured value: C, 67.72; H, 6.78.

5-O-(t-butyldiphenylsilyl)-2,3-dideoxy-2-fluoro-(L)-red sugar (erythron)-furanose (21)

In reaction flask, add lactone 20 (12.12g, 0.0325mol, 1.0eq.) and the anhydrous THF of 240mL.This solution is cooled to-78 ℃ and dripped the hexane solution of 65mL (0.065mol, 2.0eq.) 1.0 M DIBALH in 30 minutes.And-78 ℃ of lower stirrings 3 hours, after this slowly add 2.93mL (0.163mol, 5.0eq.) water stopped reaction.This reaction is warming up to room temperature and stirred 1 hour, in whole flask, form afterwards the gel transparent solid.With this reaction mixture with the dilution of the ether of two volumes and be poured in the saturated sodium tartrate aqueous solutions of potassium of the medium volume of Erlenmeyer flask.Stir 20 minutes until this emulsion is destroyed.Separate organic layer and use the 250mL ether with water layer extraction 3 times.With the organic layer dried over mgso that merges, filter, and concentrate and obtain faint yellow oily thing.This product is passed through silica gel chromatography, with 6: 1 hexane/ethyl acetate solvent systems wash-outs.The limpid oily matter of gained is obtained the white crystalline solid of 11.98g (yield 98%): R with the hexane crystallization of boiling _f(30% ether/70% hexane)=0.33; Mp 66-67 ℃. ¹H NMR (360MHz, CDCl ₃) d 7.68-7.66 (m, 4H), 7.55-7.38 (m, 6H), 5.39 (t, J=7.6Hz, 1H), 4.99 (dd, J=52.2 and 4.3Hz, 1H), 4.52 (m, 1H), 3.88 (dd, J=10.8 and 2.5Hz, 1H), 3.65 (d, J=7.9Hz, 1H), 3.49 (dd, J=7.9 and 1.8Hz, IM, (2.44-2.07 m, 2H), 1.07 (s, 9H); ¹³C NMR (100MHz, CDCl ₃) d 135.7,135.5,132.2,132.1,130.2,130.0,129.8,127.9,127.7,99.8 (d, J=31.1Hz), 96.6 (d, J=178.3Hz), 79.4,64.8,29.9 (d, J=21.2Hz), 26.8,19.2; IR (film) 3423,2932,1474,1362,1113cm ^-1HRMS theoretical value [M+Li] C ₂₁H ₂₇O ₃FSiLi:381.1874. measured value: 381.1877. ultimate analysis theoretical value .C ₂₁H ₂₇O ₃FSi:C, 67.35; H, 7.27, measured value: C, 67.42; H, 7.31.

1-O-ethanoyl-5-O-(t-butyldiphenylsilyl)-2,3-dideoxy-2-fluoro-(L)-red sugar-furanose (22)

In reaction flask, add lactonaphthol 21 (8.50g, 0.0227mol, 1.0eq.) and 170mL anhydrous methylene chloride.Then add DMAP (0.277g, 0.00277mol, 0.1eq.) and acetic anhydride (13.5mL, 0.143mol, 6.3eq.) and at room temperature stir and spend the night.React complete after, this reaction is poured in the saturated sodium bicarbonate solution.Separate organic layer, and with chloroform water layer is extracted 3 times.With the organic layer dried over mgso that merges, filter, and desolventizing obtains faint yellow oily thing.This oily matter by silica gel chromatography, is obtained 9.85g (99% yield) limpid colorless oil: R with 8: 1 hexane/ethyl acetate solvent systems wash-outs _f(30% ether/70% hexane)=0.44; ¹H NMR (360MHz, CDCl ₃) d 7.69-7.67 (m, 4H), 7.43-7.38 (m, 6H), (6.30 d, J=10.4Hz, 1H), 5.06 (d, J=54.9Hz, 1H), 4.53 (m, 1H), (3.81 dd, J=10.8 and 4.3Hz, 1H), 3.72 (dd, J=10.8 and 4.3Hz, 1H), 2.38-2.12 (m, 2H), (1.89 s, 3H), 1.07 (s, 9H); ¹³C NMR (100MHz, CDCl ₃) d 169.4,135.6,135.5,133.2,133.1,129.8,129.7,127.8,127.7,99.3 (d, J=34.1Hz), 95.5 (d, J=178.2Hz), 81.4,65.3,31.6 (d, J=20.5Hz), 26.8,21.1,19.3; IR (film) 3074,2860,1750,1589,1229,1113cm ^-1HRMS theoretical value [M-OCOCH ₃] C ₂₁H ₂₆O ₂FSi:357.1686. measured value: 357.1695. ultimate analysis theoretical value .C ₂₃H ₂₉O ₄FSi:C, 66.32; H, 7.02. measured value: C, 66.30; H, 7.04.

The exemplary process of silylanizing base and 22 couplings: (L)-5 '-O-(t-butyldiphenylsilyl)-2 ', 3-dideoxy-2 '-fluoro-5-fluorine cytidine (25)

In the flask that is equipped with short-path distillation head, add 5-flurocytosine (2.01g, 15.6mmol, 5.0eq), 35mL 1,1,1,3,3, (about 1mg) (NH of 3-hexamethyl-disilazane and catalytic amount ₄) ₂SO ₄This white suspension is heated to boiling, kept 1 hour until this alkali by silylanizing and reaction soln becomes limpid.Distill excessive HMDS, the oily resistates is vacuumized 1 hour to remove the HMDS of last tracer level.The gained white solid is dissolved in 5mL under argon atmospher anhydrous 1, the 2-ethylene dichloride.It is anhydrous 1 at 5mL to add acetic ester 22 (1.30g, 3.12mmol, 1.0eq.) in this limpid solution, the solution in the 2-ethylene dichloride.Under the room temperature to wherein adding trifluoromethanesulfonic acid trimethyl silyl ester (3.32mL, 17.2mmol, 5.5eq.).Monitor this reaction by TLC (10% methyl alcohol/90% methylene dichloride), and observe reaction when reacting 4 hours thoroughly.This reaction mixture is poured in the saturated sodium bicarbonate.Then, separate organic layer, and with chloroform water layer is extracted 3 times.With the organic layer dried over mgso that merges, filtering also, desolventizing obtains white foam.With this compound by silica gel chromatography, with 100% methylene dichloride to being present in 10% methyl alcohol in the methylene dichloride as the gradient solvent system.Isolate this compound of 1.51g (99% yield), be white foam: the mixture R of anomer _f(100%EtOAc)=0.36; Mp 74-80 ℃. ¹H NMR (400MHz, CDCl ₃) d 8.84 (bs, 1H), 8.04 (d, J=6.4Hz, 0.67H), 7.67-7.63 (m, 4H), 7.51-7.39 (m, 6.33H), 6.11 (d, J=20Hz, 0.33H), (5.98 d, J=16.4Hz, 0.67H), 5.88 (bs, 1H), 5.41 (d, J=52.4Hz, 0.33H), 5.23 (dd, J=50.4 and 4Hz, 0.67H), 4.56 (m, 0.33H), (4.45 m, 0.67H), 4.23 (dd, J=12.0 and 1.6Hz.0.67H), 3.89 (dd, J=11.2 and 3.2Hz, 0.33H), 3.74-3.66 (m, 1H), (2.45-1.96 m, 2H), 1.09 (s, 6H), 1.06 (s, 3H); ¹³C NMR (100MHz, CDCl ₃) d 158.6 (d, J=14.4Hz), 158.4 (d, J=14.4Hz), 153.9,153.8,136.6 (d, J=240.5Hz), (136.3 d, J=239.7Hz), 135.6,135.56,135.5,135.4,133.1,132.9,132.5,132.4,130.1,130.0,129.9,127.9,127.8,125.8 (d, J=33.4Hz), 124.6 (d, J=32.6Hz), 96.5 (d, J=182.0Hz), 91.7 (d, J=185.1), 90.7 (d, J=35.6Hz), 87.7 (d, J=15.2Hz), 81.5,79.5,64.9,63.0,33.5 (d, J=20.5Hz), 30.6 (d, J=20.4Hz), 26.9,26.8,19.22,19.18; IR (film) 3300,2960,1682,1608,1513,1109cm ^-1HRMS theoretical value [M+Li] C ₂₅H ₂₉N ₃O ₃SiF ₂Li:492.2106. measured value: 492.2085. ultimate analysis theoretical value .C ₂₅H ₂₉N ₃O ₃SiF ₂1/2 H ₂O:C, 60.71; H, 6.11; N, 8.50. measured value: C, 60,67; H, 6.03; N, 8.44.

The typical method of the nucleosides deprotection of silyl-protection: α-and β-(L)-2 ', 3 '-dideoxy-2 '-fluoro-5-fluorine cytidine (28a and 28b)

Nucleosides 25 (1.098g, 2.26mmol, 1.0eq.) is dissolved in the 15mL methyl alcohol, to wherein adding Neutral ammonium fluoride (0.838g, 22.6mmol, 10.0eq.).With its vigorous stirring 24 hours, TLC afterwards (15% ethanol/85% ethyl acetate) showed that this reaction finishes.This reaction mixture is filtered with the ethyl acetate dilution of 3 volumes and by little (1cm) silica gel plug.This plug is cleaned with 15% ethanol of 200mL/85% ethyl acetate solution, and desolventizing obtains white foam.By silica gel chromatography, with 15% ethanol/85% ethyl acetate solvent system wash-out, it also can separate α with the β anomer with this mixture.Obtain white foam 0.190g (0.768mmol, 34% yield), and obtain the β 0.290g (1.17mmol, 52% yield) of white foam form: (28a) R _f(15%EtOH, 85%EtOAc)=0.22; Mp 199-203 ℃ (decomposition). ¹H NMR (400MHz, CD ₃OD) d 7.78 (d, J=6.8Hz, 1H), 6.07 (d, J=19.2Hz, 1H), 5.37 (d, J=54.0Hz, 1H), 4.60 (m, 1H), 3.80 (dd, J=12.0 and 3.2Hz, 1H), 3.56 (dd, J=12.4 and 4.4Hz, 1H), 2.40-2.00 (m, 21i); ¹³C NMR (100MHZ, DMSO-d ₆) d 157.7 (d, J=13.6Hz), 153.2,135.9 (d, J=239.0Hz), (126.2 d, J=31.1Hz), 92.4 (d, J=183.6Hz), 86.7 (d, J=15.2Hz), 79.6,62.7,33.3 (d, J=20.5Hz); IR (KBr) 3343,3100,1683,1517,1104cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₁N ₃O ₃F ₂Li:254.0929. measured value: 254.0919. ultimate analysis theoretical value, C ₉H ₁₁N ₃O ₃F ₂1/2 H ₂O:C, 42.19; H, 4.72; N, 16.40. measured value: C, 42.44; H, 4.56; N, 16.56. (28b) R _f(15%EtOH, 85%EtOAc)=0.37; Mp 182-186 ℃ (decomposition). ¹H NMR (400MHz, DMSO-d ₆) d 8.32 (d, J=7.6Hz, 1H), 7.79 (bs, 1H), (7.53 bs, 1H), 5.81 (d, J=16.8Hz, 1H), 5.37 (t, J=4.8Hz), 5.18 (dd, J=51.6 and 3.2Hz, 1H), 4.32 (m, 1H), (3.88 dd, J=12.0 and 2.8Hz, 1H), 3.59 (dd, J=12.4 and 2.4Hz, 1H), 2.20-1.99 (m, 2H); ¹³C NMR (100MHz, DMSO-d ₆) d 157.7 (d, J=13.7Hz), 153.2,136.1 (d, J=237.4Hz), (125.3 d, J=33.4Hz), 97.3 (d, J=176.8Hz), 89.9 (d, J=35.7Hz), 81.6,60.2,30.3 (d, J=19.7Hz); IR (KBr) 3487,2948,1678,1509,1122cm ^-1, HRMS theoretical value [M+Li] C ₉H ₁₁N ₃O ₃F ₂Li:254.0929. measured value: 254.0935. ultimate analysis theoretical value .C ₉H ₁₁N ₃O ₃F ₂: C, 43.73; H, 4.49; N, 17.00. measured value: C, 43.69; H, 4.53; N, 16.92.

(D)-5 '-O-(t-butyldiphenylsilyl)-2 ', 3 '-dideoxy-2 '-fluoro-5-FUD (9)

The mixture R of anomer _f(1: 1 hexane/EtOAc)=0.48; Mp 65-70 ℃. ¹H NMR (400MHz, CDCl ₃) d 10.0 (bm, 1H), 7.99 (d, J=5.6Hz, 0.63H), 7.65 (m, 4H), 7.42 (m, 6.37H), 6.12 (dd, J=18.0 and 1.6Hz, 0.37H), (6.00 d, J=16Hz, 0.63H), 5.37 (dd, J=54.6 and 2.4Hz, 0.37H), 5.22 (dd, J=50.4 and 4Hz, 0.63H), 4.57 (m, 0.37H), 4.44 (m, 0.63H), 4.22 (dd, J=12.2 and 2.0Hz, 0.63H), (3.92 dd, J=11.2 and 3.2Hz, 0.37H), 3.70 (m, 1H), 2.22 (m, 2H), 1.09 (s, 5.67H), 1.074 (s, 3.33H); ¹³C NMR (100MHz, CDCl ₃) d 157.2 (d, J=31.7Hz), 157.1 (d, J=25.8Hz), 149.1,148.8,140.4 (d, J=236.6Hz), (140.1 d, J=235.2Hz), 135.6,135.5,135.4,132.9,132.7,132.4,132.3,130.1,130.0,129.9,127.9,127.8,125.1 (d, J=34.9 Hz), (123.6 d, J=34.1Hz), 96.4 (d, J=182.0 Hz), (92.0 d, J=185.9 Hz), 90.2 (d, J=37.2 Hz), (87.0 d, J=15.2Hz), 81.7,79.8,64.8,63.0,33.3 (d, J=21.2Hz), 31.0 (d, J=21.2Hz), 26.9,26.8,19.2; IR (film) 3185,1722,1117cm ^-1HRMS theoretical value [M+1] C ₂₅H ₂₉N ₂O ₄SiF ₂: 487.1866. measured value: 487.1853. ultimate analysis theoretical value C ₂₅H ₂₉N ₂O ₄SiF ₂: C, 61.71; H, 5.80; N, 5.76. measured value: C, 61.72; H, 5.86; N, 5.72.

(D)-5 '-O-(t-butyldiphenylsilyl)-2 ', 3 '-dideoxy-2 '-fluoro-5-fluorine cytidine (10)

The mixture R of anomer _f(100%EtOAc)=0.36; Mp 75-81 ℃. ¹H NMR (400MHz, CDCl ₃) d 8.50 (bm, 1H), 8.05 (d, J=6.0Hz, 0.67H), 7.67-7.63 (m, 4H), 7.51-7.39 (m, 6.33H), 6.10 (d, J=20Hz, 0.33H), (5.98 d, J=16.4Hz, 0.67H), 5.62 (bm, 1H), 5.41 (d, J=52.4Hz, 0.33H), (5.23 dd, J=51.6 and 4Hz, 0.67H), (4.57 m, 0.33H), 4.48 (m, 0.67H), 4.24 (dd, J=12.4 and 2.0Hz, 0.67H), 3.89 (dd, J=11.2 and 3.2Hz, 0.33H), 3.74-3.66 (m, 1H), (2.39-1.95 m, 2H), 1.09 (s, 6H), 1.06 (s, 3H); ¹³C NMR (100MHz, CDCl ₃) d 158.4 (d, J=14.4Hz), 158.3 (d, J=15.2Hz), 153.8,153.7,136.5 (d, J=240.5Hz), 136.2 (d, J=241.8Hz), 135.59,135.56,135.4,133.0,132.9,132.5,132.4,130.1,130.0,129.9,127.9,127.8,124.8 (d, J=31.9Hz), (96.5 d, J=181.3Hz), 91.8 (d, J=175.2Hz), (90.7 d, J=24.9Hz), 87.8 (d, J=21.2Hz), 81.6,79.6,64.9,63.0, (33.5 d, J=19.7Hz), 30.6 (d, J=21.3Hz), 26.9,26.8,19.2,14.2; IR (film) 3304,2959,1680,1621,1508,1105cm ^-1HRMS theoretical value [M+Li] C ₂₅H ₂₉N ₃O ₃SiF ₂Li:492.2106. measured value: 492.2110. ultimate analysis theoretical value .C ₂₅H ₂₉N ₃O ₃SiF ₂: C, 61.84; H, 6.02; N, 8.65. measured value: C, 61.86; H, 6.09; N, 8.55.

(D)-N ⁴-ethanoyl-5 '-O-(t-butyldiphenylsilyl)-2 ', 3 '-dideoxy-2 '-fluoro-cytidine (11)

The mixture R of anomer _f(15%EtOH, 85%EtOAc)=0.75; Mp 81-86 ℃. ¹H NMR (400MHz, CDCl ₃) d 10.58 (bs, 1H), 8.40 (d, J=7.2Hz, 0.61H), 7.86 (d, J 7.6Hz, 0.38H), (7.67-7.65 m, 4H), 7.51-7.41 (m, 6H), (7.27 d, J=8.4Hz, 1H), 6.12 (t, J 15.8Hz, 1H), 5.51 (d, J=52.6Hz, 0.38H), 5.21 (dd, J=50.8 and 2.9Hz, 0.61H), (4.62 m, 0.38H), 4.54 (m, 0.61H), 4.28 (d, J=11.5Hz, 0.61H), 3.95 (dd, J=11.9 and 3.2Hz, 0.38H), 3.79-3.70 (m, 1H), (2.46-2.04 m, 5H), 1.12 (s, 5.49H), 1.07 (s, 3.42H); ¹³C NMR (100MHz, CDCl ₃) d 171.5,171.3,163.4,154.9,144.9,144.1,135.5,135.4,133.0,132.8,132.5,132.2,130.2,130.1,129.9,128.0,127.8, (96.8 d, J=91.1Hz), 96.2 (d, J=147.9Hz), 92.3,91.2 (d, J 35.7Hz), 90.5, (88.5 d, J=15.9Hz), 81.9,80.1,64.7,62.9,33.5 (d, J=20.5 Hz), (30.5 d, J20.5 I4z), 26.9,26.8,24.9,24.8,19.3,19.2; IR (film) 3237,2932,1722,1671,1559,1493,1107cm ^-1HRMS theoretical value [M+Li] C ₂₇H ₃₂N ₃O ₄FSiLi:516.2306. measured value: 516.2310. ultimate analysis theoretical value .C ₂₇H ₃₂N ₃O ₄FSi:C, 63.63; H, 6.33; N, 8.24. measured value: C, 63.45; H, 6.42; N, 8.09.

(D)-5 '-O-(t-butyldiphenylsilyl)-2 ', 3 '-dideoxy-2 '-fluoro-cytidine (12)

The mixture R of anomer _f(15%EtOH, 85%EtOAc)=0.50; Mp 98-104 ℃. ¹H NMR (360MHz, CDCl ₃) d 7.97 (d, J=7.2Hz, 0.64H, H-6), (7.65 m, 4H), 7.47-7.38 (m, 6.36H), (6.15 d, J=20.5Hz, 0.36H), 6.05 (d, J=16.6Hz, 0.64H), 5.83 (d, J=7.9Hz, 0.36H), 5.46 (d, J=7.2Hz, 0.64H), (5.30-5.10 m, 1H), 4.55 (m, 0.36H), (4.44 m, 0.64H), 4.22 (d, J=9.7Hz, 0.64H), 3.88-3.63 (m, 1.36H), (2.38-1.95 m, 2H), 1.09 (s, 5.76H), 1.06 (s, 3.24H); ¹³C NMR (100MHz, CDCl ₃) d 166.1,155.8,141.5,140.5,135.6,135.4,133.1,132.9,132.8,132.4,130.1,130.0,129.8,128.0,127.9,127.8,96.7 (d, J=181.3Hz), 93.4 (d, J=140.3Hz), 94.5,90.8 (d, J=35.6Hz), 90.8,87.8 (d, J=15.9Hz), 81.2,79.4,65.0,63.2,33.7 (d, J=21.2Hz), 3 0.8 (d,, J=20.4Hz), 26.9,26.8,19.3,19.2:IR (film) 3470,3339,1644,1487,1113cm ^-1HRMS theoretical value [M+Li] C ₂₅H ₃₀N ₃O ₃FSiLi:474.2201. measured value: 474.2198. ultimate analysis theoretical value .C ₂₅H ₃₀N ₃O ₃FSi:C, 64.21; H, 6.47; N, 8.99. measured value: C, 64.04; H, 6.58; N, 8.76.

α-(D)-2 ', 3 '-dideoxy-2 '-fluoro-5-FUD (14a)

R _f(100％EtOAc)＝0.38；mp?153-155℃。 ¹H NMR (360MHz, CD ₃OD) d 7.80 (d, J=6.8Hz, 1H), 6.11 (d, J=18.7Hz, 1H), 5.35 (d, J=52.9,1H), 4.59 (m, 1H), 3.81 (d, J=11.9Hz, 1H), 3.57 (dd, J=12.6 and 3.6Hz, 1H), 2.36-2.15 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 159.6 (d, J=25.8Hz), 150.7,141.5 (d, J=230.6Hz), (127.0 d, J=34.9Hz), 93.9 (d, J=185.1Hz), 88.5 (d, J=15.1Hz), 81.8,64.3,34.3 (d, J=20.5Hz); IR (KBr) 3421,3081,1685,1478,1111cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₀N ₂O ₄F ₂Li:255,0769, measured value: 255.0778. ultimate analysis theoretical value C ₉H ₁₀N ₂O ₄F ₂: C, 43.56; H, 4.06; N, 11.29. measured value: C, 43.59; H, 4.11; N, 11.17.

β-(D)-2 ', 3 '-dideoxy-2 '-fluoro-5-FUD (14b)

R _f(100％EtOAc)0.54；mp?152-154℃。 ¹H NMR (360MHz, CD ₃OD) d 8.41 (d, J=7.2Hz, 1H), 5.89 (d, J 16.6Hz, 1H), 5.21 (dd, J=51.5 and 3.6Hz, 1H), 4.41 (m, 1H), 4.00 (d, J=12.6Hz, 1H), 3.67 (d, J=12.2Hz, 1H), 2.25-2.09 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 159.7 (d, J=25.8Hz), 150.7,141.8 (d, J=229.8Hz), (126.3 d, J=36.4Hz), 98.3 (d, J=179Hz), 91.9 (d, J=37.1Hz), 83.6,61.9,31.9 (d, J=20.5Hz); IR (KBr) 3417,3056,1684,1474,1105cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₀N ₂O ₄F ₂Li:255.0769. measured value: 255.0764. ultimate analysis theoretical value C ₉H ₁₀N ₂O ₄F ₂: C, 43.56; H, 4.06; N, 11.29. measured value: C, 43.37; H, 3.98; N, 11.22.

α-(D)-2 ', 3 '-dideoxy-2 '-fluoro-5-fluorine cytidine (15a)

R _f(15%EtOH, 85%EtOAc)=0.22; Mp 198-202 ℃ (decomposition). ¹H NMR (400MHz, CD ₃OD) d 7.78 (d, J=6.8Hz, 1H), 6.07 (d, J=18.8Hz, 1H), 5.37 (d, J=54.0Hz, 1H), 4.59 (m, 1H), 3.80 (dd, J=12.0 and 3.2Hz, 1H), 3.57 (dd, J=12.4 and 4.4Hz, 1H), 2.38-2.14 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 159.9 (d, J=13.6Hz), 156.5,138.3 (d, J=240.4Hz), (127.5 d, J=33.4Hz), 93.6 (d, J=184.3Hz), 89.5 (d, J=15.9Hz), 81.8,64.4,34.5 (d, J=20.5Hz); IR (Y.Br) 3486,3098,1681,1519,1108cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₁N ₃O ₃F ₂Li:254.0929. measured value: 254.0929. ultimate analysis theoretical value C ₉H ₁₁N ₃O ₃F ₂1/2 H ₂O:C, 42.19; H, 4.72; N, 16.40. measured value: C, 41.86; H, 4.75; N, 16.36.

β-(D)-2 ', 3 '-dideoxy-2 '-fluoro-5-fluorine cytidine (15b)

R _f(15%EtOH, 85%EtOAc)=0.37; Mp 181-183 ℃ (decomposition). ¹H NMR (400MHz, CD ₃OD) d 8.45 (d, J=7.2Hz, 1H), (5.92 dd, J=16.2 and 1.2Hz, 1H), (5.18 dd, J=50.9 and 4.0Hz, 114), (4.46 m, 1H), 4.05 (dd, J=12.4 and 2.4Hz, 1H), 3.72 (dd.J=12.8 and 2.4Hz, 1H), 2.27-2.05 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 159.9 (d, J=13.6Hz), 156.5,138.5 (d, J=240.5Hz), (126.9 d, J=33.4Hz), 98.4 (d, J=179.0Hz), 92.5 (d, J=36.4Hz), 83.6,61.9,31.6 (d, J=20.5Hz); IR (KBR) 3494,2944,1689,1522,1106cm ^-1HEMS theoretical value [M+Li] C ₉H ₁₁N ₃O ₃F ₂Li:254.0929. measured value: 254.0936. ultimate analysis theoretical value .C ₉H ₁₁N ₃O ₃F ₂: C, 43.73; H, 4.49; N, 17.00. measured value: C, 43.84; H, 4.47; N, 17.05.

α-(D)-N ⁴-ethanoyl-2 ', 3 '-dideoxy-2 '-fluoro-cytidine (16a)

R _f(15％EtOH，85％EtOAc)＝0.40；mp?208-212℃。 ¹H NMR (360 MHz, DMSO-d ₆) d (10.91, bs, 1H), 8.05 (d, J=7.2Hz, 1H), 7.25 (d, J=7.2Hz, 1H), 6.08 (dd, J=19.1 and 2.9Hz, 1H), 5.42 (d, J=52.2Hz, 1H), (4.97 bs, 1H), 4.54 (m, 1H), 3.63 (d, J=13.0Hz, 1H), 3.47 (d, J=13.3Hz, 1H), (2.35-2.15 m, 2H), 2.11 (s, 3M; ¹³C NMR (100MHz, DMSO-d ₆) d 171.0,162.6,154.3,145.7,94.9,92.0 (d, J=183.6Hz), 87.5 (d, J=15.9Hz), 80.2,62.6,33.3 (d, J=19.7Hz), 24.4; IR (KBr) 3436,3227,1702,1661,1442,1102cm ^-1HRMS theoretical value [M+Li] C ₁₁H ₁₄N ₃O ₄FLi:278.1128. measured value: 278.1136. ultimate analysis theoretical value C ₁₁H ₁₄N ₃O ₄F:C, 48.71; H, 5.20; N, 15.49. measured value: C, 48.73; H, 5.23; N, 15.52.

β-(D)-N ⁴-ethanoyl-2 ', 3 '-dideoxy-2 '-fluoro-cytidine (16b)

R _f(15％EtOH，85％EtOAc)＝0.50；mp?174-178℃。 ¹HNMR (360MHz, DMSO-d ₆) d (10.90, bs, 1H), 8.46 (d, J=7.2Hz, 1H), 7.18 (d, J=7.2Hz, 1H), 5.90 (d, J=16.9Hz, 1H), 5.27 (d, J=52.9Hz, 1H), (5.27 bs, 1H), 4.39 (m, 1H), 3.88 (d, J=13.0Hz, 1H), 3.61 (d, J=13.0Hz, 1H), (2.09 s, 3H), 2.20-1.85 (m, 2H); ¹³C NMR (100MHz, DMSO-d ₆) d 171.0,162.6,154.4,144.7,97.0 (d, J=177.5Hz), 95.0,90.7 (d, J=36.6Hz), 82.2,60.3,30.3 (d, J=19.7Hz), 24.3; IR (KBr) 3447,3245,1703,1656,1497,1122cm ^-1HRMS theoretical value [M+Li] C ₁₁H ₁₄N ₃O ₄FLi:278.1128. measured value: 278.1133. ultimate analysis theoretical value C ₁₁H ₁₄N ₃O ₄F:C, 48.71; H, 5.20; N, 15.49. measured value: C, 48.65; H, 5.22; N, 15.46.

α-(D)-2 ', 3 '-dideoxy-2 '-fluoro-cytidine (17a)

R _f(15%EtOH, 85%EtOAc)=0.08; Mp 234-237 ℃ (decomposition). ¹H NMR (400MHz, DMSO-d ₆) d 7.52 (d, J=7.6Hz, 1H), 7.21 (bm, 2H), (6.05 dd, J=20.4 and 3.2Hz, 1H), 5.73 (d, J=7.2Hz, 1H), 5.28 (d, J=52.4Hz, 1H), 4.93 (t, J=5.6Hz, 1H), (4.45 m, 1H), 3.58 (m, 1H), (3.43 m, 1H), 2.26-2.13 (m, 2H); ¹³C NMR (100MHz, DMSO-d ₆) d 165.8,155.0,141.6,93.3,92.2 (d, J=182.8Hz), 86.6 (d,

J=15.1Hz), 79.4,62.8,33.3 (d, J=19.7Hz); IR (KBr) 3366,3199,1659,1399,1122cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₂N ₃O ₃FLi:236.1023. measured value: 236.1014. ultimate analysis theoretical value C ₉H ₁₂N ₃O ₃F:C, 47.16; H, 5.28; N, 18.33. measured value: C, 47.40; H, 5.34; N, 18.51.

β-(D)-2 ', 3 '-dideoxy-2 '-fluoro-cytidine (17b)

Nucleosides 25 (0.160g, 0.59mmol) is dissolved in the methanol solution of 10mL saturated ammonia.Stir after 5 minutes, react complete.Remove the ammonia solution of methyl alcohol and the gained white solid is placed under the vacuum, mild heat 2 hours is to remove the ethanamide by product by distillation in 60 ℃ of water-baths.With the crystallization from 5% methyl alcohol/95% methylene dichloride of this white solid, obtain the white crystalline solid of quantitative yield.R _f(15%EtOH, 85%EtOAc)=0.18; Mp 191-195 ℃ (decomposition). ¹H NMR (360MHz, CD ₃OD) d 8.10 (d, J=7.2Hz, 1H), 5.92 (d, J=17.3Hz, 1H), 5.82 (d, J=7.6Hz, 1H), 5.13 (d, J=50.0Hz, 1H), (4.39 m, 1H), 3.97 (d, J=12.2Hz, 1H), 3.68 (dd, J=13.0 and 2.5Hz, 1H), 2.21-2.00 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 165.9,155.0,140.8,97.3 (d, J=176.8Hz), 93.6,90.3 (d, J=35.6Hz), 81.3,60.7,31.0 (d, J=20.5Hz); IR (KBr) 3397,3112,1680,1400,1178,1070cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₂N ₃O ₃FLi:236.1024. measured value: 236.1028. ultimate analysis theoretical value .C ₉H ₁₂N ₃O ₃F:C, 47.16; H, 5.28; N, 18.33. measured value: C, 47.01; H, 5.21; N, 18.29.

(L)-5 '-O-(t-butyldiphenylsilyl)-2 ', 3 '-dideoxy-2 '-fluoro-thymidine (23)

The mixture R of anomer _f(10%MeOH/90% methylene dichloride)=0.56; Mp 61-65 ℃. ¹H NMR (360, MHz, CDCl ₃) d 9.48 (bs, 1H), 7.67 (m, 4H), (7.45-7.37 m, 7H), 6.15 (dd, J=20.2 and 3.2Hz, 0.36H), 5.99 (d, J=18.4Hz, 0.64H), (5.34 d, J=51.8Hz, 0.36H), 5.24 (dd, J=52.2 and 4.3Hz, 0.64H), 4.59 (m, 0.36H), (4.45 m, 0.64H), 4.17 (dd, J=12.2 and 2.5Hz, 0.64H), 3.91 (dd, J=11.9 and 2.9Hz, 0.36H), (3.81 dd, J=11.5 and 2.9Hz, 0.64H), 3.68 (dd, J=10.8 and 3.6Hz, 0.36H), 2.40-2.12 (m, 2H), 1.94 (s, 1.08H), (1.61 s, 1.92H), 1.10 (s, 5.76H), 1.07 (s, 3.24H); ¹³C NMR (100MHz, CDCl ₃) d 164.1,164.0,150.4,150.2,136.4,135.6,135.5,135.4,135.3,135.2,133.0,132.8,132.6,130.1,130.0,129.9,127.94,127.90,127.8,110.8,109.8,96.4 (d, J=181.3Hz), (92.1 d, J=185.8Hz), 90.7 (d, J=36.4Hz), (86.6 d, J=15.2Hz), 80.9,79.4,64.9,63.6,33.4 (d, J=20.5Hz), (32.0 d, J=21.2Hz), 27.0,26.8,19.4,19.2,12.6,12.2; IR (film) 3183,3050,1696,1506,1188cm ^-1HRMS theoretical value [M+Li] C ₂₆H ₃₁N ₂O ₃SiF:489.2197. measured value: 489.2175. ultimate analysis theoretical value C ₂₆H ₃₁N ₂O ₃SiF:C, 64.71; H, 6.47; N, 5.80. measured value: C, 64.88; H, 6.56; N, 5.76.

(L)-5 '-O-(t-butyldiphenylsilyl)-2 ', 3 '-dideoxy-2 '-fluoro-5-FUD (24)

The mixture R of anomer _f(1: 1 hexane/EtOAc)=0.48; Mp 65-71 ℃. ¹H NMR (400MHz, CDCl ₃) d 9.08 (bs, 0.4H), 9.00 (bs, 0.6H) 8.01 (d, J=5.4Hz, 0.6H), 7.65 (m, 4H), (7.42 m, 6.4H), 6.10 (dd, J=20.2 and 1.4Hz, 0.4H), 6.00 (d, J=16.0Hz, 0.6H), 5.3 5 (dd, J=52.4 and 1.6Hz, 0.4H), (5.22, dd, J=51.2 and 4Hz, 0.6H), (4.57 m, 0.4H), 4.44 (m, 0.6H), 4.22 (dd, J=12.4 and 2.0Hz, 0.6H), 3.91 (dd, J=11.2 and 2.9Hz, 0.4H), 3.70 (m, 1H), (2.45-2.00 m, 2H), 1.09 (s, 5.4H), 1.07 (s, 3.6H); ¹³C NMR (100MHz, CDCl ₃) d 156.9 (d, J=26.5Hz), 148.8,148.6,140.3 (d, J=236.7Hz), 140.1 (d, J=235.1Hz), 135.6,135.5,135.4,132.9,132.7,132.4,132.3,130.2,130.1,129.9,127.9,127.8,125.1 (d, J=34.9Hz), 123.6 (d, J=34.2Hz), 96.4 (d, J=182.9Hz), 92.0 (d, J=186.6Hz), 90.2 (d, J=36.0Hz), 86.9 (d, J=15.1Hz), 81.7,79.8,64.8,63.0,33.2 (d, J=20.5Hz), 30.9 (d, J=20.4Hz), 26.9,26.8,19.2; IR (film) 3191,1719,1113cm ^-1HRMS theoretical value [M+Li] C ₂₆H ₃₁N ₂O ₃SiF Li:493.1946. measured value: 493.1952. ultimate analysis theoretical value C ₂₆H ₃₁N ₂O ₃SiF:C, 61.71; H, 5.80; N, 5.76. measured value: C, 61.73; H, 5.83; N, 5.77.

α-(L)-2 ', 3 '-dideoxy-2 '-fluoro-thymidine (26a)

R _f(100％EtOAc)＝0.25；mp?147-149℃。 ¹H NMR (360MHz, CD ₃OD) d 7.45 (s, 1H), 6.11 (dd, J=19.4 and 2.9Hz, 1H), 5.30 (d, J=53.6Hz, 1H), (4.58 m, 1H), 3.79 (dd, J=12.2 and 2.2Hz, 1H), 3.55 (dd, J=2.2 and 3.6Hz, 1H), 2.40-2.15 (m, 2H), 1.87 (s, 3H); ¹³C NMR (100MHz, CD ₃OD) d 166.6,152.3,138.6,110.5,93.9 (d, J=185.1Hz), 88.3 (d, J=15.1Hz), 81.7,64.4,34.5 (d, J=20.5Hz), 12.6; IR (KBr) 3436,3166,1727,1667,1362,1186c ^-1HRMS theoretical value [M+Li] C ₁₀H ₁₃N ₂O ₄FLi:251.1019. measured value: 251.1014. ultimate analysis theoretical value C ₁₀H ₁₃N ₂O ₄F:C, 49.18; H, 5.37; N, 11.47. measured value .C, 49.32; H, 5.40; N, 11.29.

β-(L)-2 ', 3 '-dideoxy-2 '-fluoro-thymidine (26b)

R _f(100％EtOAc)＝0.38；mp?186-188℃。 ¹H NMR (360MHz, CD ₃OD) d 7.94 (s, 1H), 5.93 (d, J=17.6Hz, 1H), 5.20 (d, J=51.8Hz, 1H), (4.40 m, 1H), 3.98 (d, J=11.9Hz, (H), 3.68 (d, J=13.0Hz, 1H), 2.37-2.10 (m, 2H), 1.83 (s, 3H); ¹³C NMR (100MHz, CD ₃OD) d 166.7,152.3,138.2,111.0,98.4 (d, J=178.3Hz), 92.1 (d, J=36.4Hz), 83.1,62.4,32.5 (d, J=20.5Hz), 12.6; IR (KBr) 3478,3052,1684,1363,1192,1005cm ^-1Ultimate analysis theoretical value C ₁₀H ₁₃N ₂O ₄F:C, 49.18; H, 5.37; N, 11.47. measured value: C, 49.29; H, 5.44; N, 11.36.

α-(L)-2 ', 3 '-dideoxy-2 '-fluoro-5-FUD (27a)

R _f(100％EtOAc)＝0.38；mp?155-157℃。 ¹H NMR (400MHz, CD ₃OD) d 7.80 (d, J=6.8Hz, 1H), (6.13 d, J=20.0Hz, 1H), (5.35 d, J=54.4Hz, 1H), 4.63 (m.1H), 3.81 (dd, J=11.9 and 3.2Hz, 1H), 3.58 (dd, J=12.4 and 2.0Hz, 1H), 2.41-2.15 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 159.6 (d, J=25.8Hz), 150.7,141.5 (d, J=230.6Hz), (127.0 d, J=34.9Hz), 93.9 (d, J=184.3Hz), 88.5 (d, J=15.1Hz), 81.9,64.3,34.3 (d, J=20.5Hz); IR (KBr) 3401,3098,1661,1458,1018cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₀N ₂O ₄F ₂Li:255.0769. measured value: 255.0771. ultimate analysis theoretical value C ₉H ₁₀N ₂O ₄F ₂: C, 43.56; H, 4.06, N, 11.29. measured value: C, 43.70; H, 4.17; N, 11.15.

β-(L)-2 ', 3 '-dideoxy-2 '-fluoro-5-FUD (27b)

R _f(100％EtOAc)＝0.54；mp?153-156℃。 ¹H NMR (400MHz, CD ₃OD) d 8.46 (d, J=6.8Hz, 1H), 5.94 (d, J=16.4Hz, 1H), 5.25 (dd, J=51.6 and 4.0Hz, 1H), 4.41 (m, 1H), 4.05 (dd, J=12.8 and 2.4Hz, 1H), 3.72 (dd, J=12.4 and 2.4 Hz, 1H), 2.34-2.09 (m, 2H); ¹³C NMR (100MHz, CD ₃OD) d 159.7 (d, J=25.8Hz), 150.7,141.8 (d, J=230.6Hz), (126.3 d, J=35.7Hz), 98.3 (d, J=184.6Hz), 91.9 (d, J=36.4Hz), 83.6,61.9,31.9 (d, J=20.5Hz); IR (KBr) 3482,3037,1702,1654,1402,1103cm ^-1HRMS theoretical value [M+Li] C ₉H ₁₀N ₂O ₄F ₂Li:255.0769. measured value: 255.0764. ultimate analysis theoretical value C ₉H ₁₀N ₂O ₄F ₂: C, 43.56; H, 4.06; N, 11.29. measured value: C, 43.59; H, 4.06; N, 11.17.

Preparation L-2 '-fluoro-2 ', 3 '-unsaturated nucleosides

The second of unsaturated 2 '-fluoro nucleosides is easy to synthetic method and now finishes and be described below.This is synthetic to comprise protected pyrimidine or purine bases and key intermediate 309 is reacted in the presence of Lewis acid, total as described in the following scheme 9.The typical compound synthetic according to this synthetic method sees Table 5-6.

Scheme 9

Reagent; (i) 2-methoxyl group propylene, DMF, p-TsOH (ii) NalO ₄, H ₂O (iii) (EtO) ₃P (O) CHFCO ₂Et.NaHMDS, THF ,-78 ℃ of (iv) C-HCl.EtOH (v) TBDMSCl, imidazoles, CH ₂Cl ₂(vi) DIBAL-H, CH ₂Cl ₂,-78 ℃ of (vii) Ac ₂O, pyridine, CH ₂Cl ₂,

Scheme 9

Reagent: (i) silylanizing uridylic, TMSOT1, DCE (ii) silylanizing thymus pyrimidine, TMSOT1, DCE (111) silylanizing N ⁴-Bz-cytosine(Cyt), TMSOTI, CH ₃CN (iv) 5-F-cytosine(Cyt), TMSOTI, CH ₃CN (v) TBAF, CH ₃CN (vi) NH ₃/ MeOH

Scheme 9

Reagent: (i) silylanizing 6-Cl, purine, TMSOTI, DCE (ii) silylanizing s-Cl-2-F-purine, TMSOTI, DCE (iii) TBAF, CH ₃CN (iv) NH ₃/ DME (v) NH ₃/ MeOH, 90 ℃ of (vi) HSCH ₂CH ₃OH, MeOMe, MeOH refluxes,

Table 5

As previously mentioned, finished the synthetic of 2 ', 3 '-unsaturated-D-nucleosides by the elimination reaction that is begun by the facile nucleoside analog of appearance, comprising the tediously long modification reaction of single core thuja acid.Some reported by suitable 2 '-elimination reaction preparation D-2 '-fluoro-2 ', 3 '-unsaturated pyrimidine nucleoside (Martin, J.A. etc., J.Med Chem.1990,33,213 7-2145 of fluorinated nucleoside analogue; Stezycki, R.Z. etc., J.Med.Chem.1990,33,2150-2157).But during with the synthetic L-Fd4N of this method, in preparation initiator L-nucleosides, run into the other difficulty.Because 2, the unsaturated sugar moieties of 3-is unstable in the presence of Lewis acid under coupling condition, by direct polycondensation synthetic 2 ', 3 '-example of unsaturated purine nucleoside is considerably less, the situation thiophene phenyl intermediate (Abdel-Medied that only has a kind of pyrimidine analogue, A.W.S. etc., Synthesis 1991,313-317; Sujino, K. etc., Tetrahedron Lett.1996,37,6133-6136).Opposite with the unsaturated sugar moieties of 2,3-, 2-fluoro-2, the unsaturated sugar of 3-glycosyl bond stability with the heterocycle condensation time improves, and estimates can become more stable to direct linked reaction.Therefore, select (R)-2-fluoro butenolide 506, as the precursor of key intermediate 508, they can be by L-Glycerose acetonide (glyceraldehyde acetonide) 501 preparations.

Begun by L-Glycerose acetonide, by Horner-Emmons reaction in the presence of alpha-fluoro phosphoryl acetic acid triethyl ester and two (trimethyl silyl) ammonification sodium, in THF, obtain (E)-502/ (Z)-2 mixture ( ¹H NMR detects 9: 1) (Thenappan, A. etc., J.Org.Chem., 1990,55,4639-4642; Morikawa, T. etc., Chem.Pharm.Bull.1992,40,3189-3193; Patrick, T.B. etc., J.Org.Chem.1994,59,1210-1212).Because the difficulty in separating (E)-502/ (Z)-502 isomer is used for following ring-closure reaction with this mixture, obtains the glycol 504 of required lactone 503 and not cyclization under acidic conditions.Change the gained mixture into silylated lactone 506, it is obtained lactonaphthol 507-78 ℃ of lower reduction with DIBAL-H in methylene dichloride.This lactonaphthol 507 usefulness acetic anhydride processing is obtained key intermediate 508, itself and the condensation under the Vorburggen condition of silylated 6-chloro-purine are obtained anomer mixture 509.In THF, process 509 nucleosides that obtain dissociating 510 and 511 with TBAF, easily be isolated with silica gel chromatography.By in stainless steel cylinder, respectively obtaining Adenine derivatives 512 and 513 at 90 ℃ of lower compounds 510 and 511 of processing with mercaptoethanol and NaOMe.Process compound 510 and 511 with mercaptoethanol and NaOMe and obtain respectively inosine analogue 514 and 515.The stereochemistry of these compounds is determined (having the intersection peak in the B isomer 512 between H-1 ' and H-4 ') by the NOESP wave spectrum.

Scheme 10. is by the synthetic L-2 ' of direct polycondensation-fluoro-d4 VITAMIN B4 and xanthoglobulin

Reagent: (i) (EtO) ₂P (O) CHFCO ₂Et, [(CH ₃) ₃Si] ₂NNa, THF ,-78 ℃ of (ii) HCVEtOH (iii) TBDMSCl, imidazoles, CH ₂Cl ₂(iv) 1 M DIBAL-H is in CH ₂Cl ₂, CH ₂Cl ₂,-78 ℃ of (v) Ac ₂O, pyr., CH ₂Cl ₂(vi) silylanizing 6-Cl-purine, TMSOTl, DCE (vii) TBAF, CH ₃CN (viii) NH ₃/ MgOH, 90 ℃ of (ix) HS (CH ₂) ₂OH, NaOMe/MeOH refluxes

Half virtual value (the EC of table 7. L-2 '-fluoro-d4 VITAMIN B4 and xanthoglobulin anti-HIV-1 in PBM ₅₀) and half inhibiting value (IC ₅₀)

Experimental section

Detecting fusing point with Mel-temp II experimental installation does not calibrate.Record NMR (Nuclear Magnetic Resonance) spectrum as internal standard substance with Bruker 250 and AMX400 400MHz chromatographic instrument with the tetramethyl-silicomethane; Chemical shift (δ) with 1,000,000/report (ppm), and signal description is s (unimodal), d (doublet), t (triplet), q (quartet), br s (wide unimodal), dd (doublet of doublet) and m (multiplet).UV spectrum derives from beckman DU 650 spectrographs.Opticity is measured at the digital polariscope of Jasco DIP-370.Mass spectrum detects at Micromass Inc.Autospec high resolution double focussing sector (EBE) MS spectrograph.Infrared spectra is recorded in Nicolet 510 FT-IR spectrographs.By Atlantic Mcrolab, Inc., Norcross, GA carries out ultimate analysis.Institute responds and uses thin-layer chromatography Analtech, 200mm silica gel G F board monitoring.Use before use Ca ₂The H distillation obtains 1 of drying, 2-ethylene dichloride, methylene dichloride and acetonitrile.With sodium and benzophenone distillation, when the solution purpling, obtain dry THF.

L-(S)-Glycerose acetonide (302)

DMF (1L) solution of L-GuA-gamma lactone (175g, 0.98mol) is cooled to 0 ℃ and under agitation add tosic acid (1.1g, 5.65mmol) in batches.In gained solution, under 0 ℃, drip 2-methoxyl group propylene (87.7g, 0.92mol) by dropping funnel.This reaction mixture was warming up to room temperature and restir 24 hours.React complete after, add yellow soda ash (124g) and with gained suspension vigorous stirring 3 hours.Then, obtain yellow residue with the glass filter filtration and with this filtrate vacuum-evaporation, add toluene (170mL), crystallization occurs.The suction filtration solid is used hexane/ethanol (9: 1; 1L) washing, and drying obtains faint yellow solid 301 (99.1g, 65%).

To stir, 5,6-O-isopropylidene-L-GuA-1,4-lactone (70.0g, 0.32mol) in the suspension in water (270mL), in 30 minutes, adding sodium metaperiodate (123g under 0 ℃ in batches, 0.58mol), keep pH5.5 (regulating by adding 2N sodium hydroxide).To stir 2 hours under this suspension room temperature, then and filtration saturated with sodium-chlor.With the pH regulator of this filtrate to 6.5-7.0, and with methylene dichloride (5 times, 200mL) and ethyl acetate (5 times, 300mL) extraction.With anhydrous magnesium sulfate that the organic layer that merges is dry, filter and concentrating under reduced pressure (＜20 ℃).Then the distillation of gained resistates is obtained 302 (23.2g, 69%), be colorless oil.b.p.49-51℃/16Torr。[α] _D(25-66.4 c 6.3, benzene).

(E)/(Z)-ethyl-3-[(R)-2,2-dimethyl-1,3-dioxolane-4-yl]-2-fluorinated monomer (E-303 and Z-303)

THF (70mL) solution of 2-fluorine phosphoryl triethyl acetate (39.2g, 162mmol) is cooled to-78 ℃, and drips two (trimethyl silyl) ammonification sodium (1.0 M THF solution, 162mL, 162mmol).This mixture was kept 30 minutes at-78 ℃, then add THF (70mL) solution of 303 (19.14g, 147mmol).-78 ℃ of lower stirrings after 1 hour are processed this reaction mixture and are used extracted with diethyl ether with aqueous ammonium chloride solution.With saturated sodium-chloride washing ether phase, use dried over mgso, filter and evaporation.This resistates is obtained E-303 and Z-303 (9: 1 bases in the enterprising circumstances in which people get things ready for a trip spectrum of silica gel ¹H NMR), be faint yellow oily thing (34.6g, 97.9%). ¹HNMR(CDCl ₃)81.34，1.36(2t，J＝8Hz，-CH ₂CH ₃)，1.40，1.45(2s，-?CH ₃)，3.69(m，H _a-5)，4.28(m，H _b-5，-CH ₂CH ₃)，5.02(m，H-4)，5.40(m，H-4)，6.02(dd，J＝8，20Hz，H-3)，6.18(dd，J＝8，32Hz，H-3)。

(R)-(+)-and 4-[(t-butyldimethylsilyloxy base) methyl]-2-fluoro-2-butylene-4-lactone (307)

The 110mL ethanol solution of E-303 and Z-303 (19.62g, 89.89mmol) is processed and at room temperature stirred 2 hours with the 30mL concentrated hydrochloric acid.Solvent removed in vacuo and with this resistates and toluene (3 * 300mL) coevaporations obtain the ester 305 of lactone 304 and not cyclization.The yellowish mill base of gained is directly used in the next step without purifying.To 304,305 and the mixture of imidazoles (12.3g, 180mmol) in methylene dichloride (250mL) in add tert-butyldimethylsilyl chloride (27.1g, 180mmol), and will be under this reaction mixture room temperature stir 4 hours.The gained mixture is washed with water, use dried over mgso, filter and be concentrated into dried.Separate this resistates with the 4%EtOAc-hexane as eluent by silica gel column chromatography, obtain 307 (28.0g, 70.2%, by compound 302), be white crystalline solid.Mp48-50 ℃; [α] _D+ 105.3 (c1.60, CHCl ₃); ¹H NMR (CDCl ₃) δ 0.07,0.08 (2s, 2xCH ₃), 0.88 (s, the tertiary butyl), 3.88 (m, 2H, H-5), 5.01 (m, 1H, H4), 6.73 (ps t, 1H, J=4Hz); Ultimate analysis theoretical value C ₁₀H ₁₉FO ₃Si:C, 53.63; H, 7.77. measured value: C, 53.70; H, 7.75.

1-ethanoyl-4-[(t-butyldimethylsilyloxy base) methyl]-2-fluoro-2-butylene-4-lactone (309)

Under nitrogen atmosphere, lactone 307 (20.59g, 83.54mmol) is dissolved in the methylene dichloride of 200mL, then be cooled to-78 ℃, and add methylene dichloride (125mL) solution of 1.0M DIBAL-H.The gained compound was stirred 2 hours under-78 ℃.With this cold mixture of rare nitric acid treatment, and dry (sodium sulfate).Solvent evaporation is obtained anomer 308, for faint yellow oily thing (16.6g, thick yield 80%), with its not purified the next step that is directly used in.

At 0 ℃ with Ac ₂O (25mL, 0.27mol) join 308 and methylene dichloride (200mL) solution of pyridine (22mL, 0.27mol) in, and the gained mixture stirred 16 hours.With dilute hydrochloric acid, saturated sodium bicarbonate and this reaction mixture of salt water washing.The organic layer that merges is dry, filter and be concentrated into dried.This resistates is carried out column chromatography purifying (6.5% ethyl acetate/hexane), obtain 309 (12.6g, 65%), be colorless oil.

The general method of acetic ester 309 and pyrimidine base condensation

Under nitrogen atmosphere, with uridylic (420mg, 3.75mmol), the mixture of hexamethyl-disilazane (15mL) and ammonium sulfate (20mg) refluxed 3 hours.Gained clear solution vacuum concentration is extremely done.In the solution of the DCE of drying (20ml), under 0 ℃, add TMSOTf (0.7mL, 3.14mmol) to sugar 309 (728mg, 2.50mmol) and silyl choline.This reaction mixture was stirred 2 hours under nitrogen atmosphere, pour in the cold saturated sodium bicarbonate solution (30mL) and stirred 15 minutes.With the washing of gained mixture, dry (sodium sulfate) filters and vacuum concentration.This crude product is obtained 310 (0.960g, 2.73mmol, 73%) by column chromatography (3% methyl alcohol/chloroform) purifying, for inseparable anomer, it is directly used in the next step without separation.

1-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-L-glyceryl-penta-2-alkene furyl glycosyl] uridylic (310)

UV (CHCl ₃) λ _Max257.5nm.; Ultimate analysis (C ₁₅H ₂₃N ₂O ₄Si) C, H, N.

1-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-L-glyceryl-penta-2-alkene furyl glycosyl] thymus pyrimidine (311)

Silylated thymus pyrimidine (242mg, 1.92mmol), 307 (500mg, 1.72mmol) and TMSOTf (0.5mL, 2.25mmol) reaction obtained 311 mixture in 2 hours, be inseparable anomer mixture (0.392g, 1.10mmol, 64%) with it by silica gel column chromatography (3% methyl alcohol/chloroform) purifying.UV(CHCl ₃)λmax262.0nm。Ultimate analysis (C ₁₆H ₂₅FN ₂O ₄Si) C, H, N.

N ⁶-benzoyl-1-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-(a, b)-L-glyceryl-penta-2-alkene furyl glycosyl] cytosine(Cyt) (312 and 313)

Silylated N ⁶-benzoyl cytosine(Cyt) (790mg, 3.67mmol), 307 (470mg; 1.62mmol) and TMSOTf (0.5mL; 2.25mmol) reaction obtained 312 and 313 mixture in 2 hours, and it is obtained β anomer 312 (0.34g, 0.76mmol by silicagel column (30% ethyl acetate/hexane) purifying; 47.1%); be white solid, and α anomer 313 (0.23g, 0.52mmol; 31.8%), is white solid.312:UV (CHCl ₃) λ max260.5nm; Ultimate analysis (C ₂₂H ₂₈FN ₃O ₄Si) C, H, N.; 513:UV (CHCl ₃) λ _Max260.5nm.; Ultimate analysis (C ₂₂H ₂₈FN ₃O ₄Si) C, H, N.

5-fluoro-1-[5-O-(t-butyldimethylsilyl l-2, and 3-dideoxy-2-fluoro-(a, b-L-glyceryl-penta-2-alkene furyl glycosyl] cytosine(Cyt) (314 and 315)

Silylated 5-fluoro-cytosine(Cyt) (300mg, 2.32mmol), 309 (360mg, 1.24mmol) and TMSOTf (0.4mL, 1.86mmol) reaction obtained 314 and 315 mixture in 2 hours, it is obtained β anomer 314 white solids (168mg, 37.8%) and α anomer 315 (121mg, 27.1%) white solid by silica gel column chromatography (3%MeOH/ methylene dichloride) purifying.314：UV(MeOH)λ _max281.5nm；315：UV(MeOH))λ _max281.5mn。

1-(2,3-dideoxy-2-fluoro-(α, β)-L-glyceryl-penta-2-alkene furyl glycosyl) uridylic (316 and 317)

To fluoridize that tetra-n-butyl ammonium (0.6mL, 0.6mmol) joins in the mixture of 310 (177mg, 0.52mmol) in THF (15mL) and will stir 15 minutes under this reaction mixture room temperature.Desolventizing and with this resistates by silica gel column chromatography (2%MeOH/CHCl ₃) purifying obtains β anomer 316 (52.8mg, 0.23mmol, 44.5%) and α anomer 317 (35.1mg, 0.15mmol, 29.6%).

316:UV (H ₂O) λ _Max261.0mn (pH7); Ultimate analysis (C ₉H ₉FN ₂O ₄0.3H ₂O) C, H, N.317:UV (H ₂O) λ _Max261.0nm (pH7); Ultimate analysis (C ₉H ₉FN ₂O ₄0.2H ₂O) C, H, N.

1-(2,3-dideoxy-2-fluoro-(α, β)-L-glyceryl-penta-2-alkene furyl) thymus pyrimidine (318 and 319)

At 0 ℃, will fluoridize tetra-n-butyl ammonium (0.8mL, 0.8mmol) and join in the mixture of 311 (240mg, 0.67mmol) in THF (10mL), and will stir 15 minutes under this reaction mixture room temperature.Desolventizing also obtains β anomer 318 (66.5mg, 0.28mmol, 41%) and α anomer 319 (52.8mg, 0.23nunol, 26%) with this resistates by silica gel column chromatography (40%THF/ hexanaphthene) purifying.

318:UV (H ₂O) λ _Max265.5nm (pH7); Ultimate analysis (C ₉H ₉FN ₂O ₄0.3H ₂O) C, H, N.319:UV (H ₂O) λ _Max266.0nm (pH7); Ultimate analysis (C ₉H ₉FN ₂O ₄0.3H ₂O) C, H, N.

N ⁶-benzoyl-1-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) cytosine(Cyt) (320)

To fluoridize in THF (10mL) solution that tetra-n-butyl ammonium (1M is in THF) (1mL, 1mmol) joins β anomer 312 (280mg, 0.63mmol) and with this reaction and at room temperature stir 1 hour.With this reaction mixture concentrating under reduced pressure and with this resistates by flash silica gel column 2.5%MeOH/CHCl ₃The wash-out purifying obtains 320 (218mg, 0.66mmol, 75%), is white solid.

UV(MeOH)λ _max260.5nm。Ultimate analysis (C ₁₆H ₁₄FN ₃O ₄) C, H, N.

N ⁶-benzoyl-1-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) cytosine(Cyt) (321)

To fluoridize in THF (10mL) solution that tetra-n-butyl ammonium (1M is in THF) (1mL, 1mmol) joins α anomer 313 (280mg, 0.63mmol) and with this reaction and at room temperature stir 1 hour.With this reaction mixture concentrating under reduced pressure and with this resistates by flash silica gel column 2.5%MeOH/CHCl ₃The wash-out purifying obtains 321 (145.8mg, 0.44mmol, 69%), is white solid.

UV(MeOH)λ _max260.5nm。Ultimate analysis (C ₁₆H ₁₄FN ₃O ₄0.3H ₂O) C, H, N.

1-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) cytosine(Cyt) (322)

Methyl alcohol (5mL) solution NH with β anomer (67.60mg, 0.204mmol) ₃/ MeOH (10mL saturated solution) processes and will stir under this reaction mixture room temperature until observe initiator disappearance (10 hours).Pass through preparation TLC purifying with this reaction mixture concentrating under reduced pressure and with this resistates, make eluent with the 12%MeOH/ methylene dichloride.The material 322 (43mg, 93.1%) of plate acquisition is solid thus, and it is ground with hexane and ether.

UV (H ₂O) λ _Max266.5nm (pH7); Ultimate analysis (C ₉H ₁₀FN ₃O ₃.0.4H ₂O) C, H, N.

1-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) cytosine(Cyt) (323)

Methyl alcohol (5mL) solution NH with α anomer (65.90mg, 0.199mmol) ₃/ MeOH (10mL saturated solution) processes and will stir under this reaction mixture room temperature until observe initiator disappearance (16 hours).Pass through preparation TLC purifying with this reaction mixture concentrating under reduced pressure and with this resistates, make eluent with the 12%MeOH/ methylene dichloride.The material 323 (42.5mg, 94.5%) of plate acquisition is solid thus, and it is ground with hexane and ether.

UV (H ₂O) λ _Max276.0nm (pH7); Ultimate analysis (C ₉H ₁₀FN ₃O ₃) C, H, N.

5-fluoro-1-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) cytosine(Cyt) (324)

To fluoridize in the acetonitrile solution that tetra-n-butyl ammonium (1M is in THF) joins β anomer 314 and will stir 1 hour under this reaction room temperature.With this reaction mixture concentrating under reduced pressure and with this resistates by flash silica gel column 12%MeOH/CHCl ₃The wash-out purifying obtains 324.

5-fluoro-1-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) cytosine(Cyt) (325)

To fluoridize in the acetonitrile solution that tetra-n-butyl ammonium (1M is in THF) joins α anomer 315 and will stir 1 hour under this reaction room temperature.With this reaction mixture concentrating under reduced pressure and with this resistates by flash silica gel column 12%MeOH/CHCl ₃The wash-out purifying obtains 325.

The general method of acetic ester 309 and purine bases condensation

Under nitrogen atmosphere, with 6-chloropurine (1.20g, 7.75mmol), the mixture of hexamethyl-disilazane (25mL) and ammonium sulfate (catalytic amount) refluxed 4 hours.Gained clear solution vacuum concentration to dry doubling is dissolved in this resistates among the anhydrous DCE (10mL) and at room temperature with 307 (1.50g, 5.17mmol) DCE (40mL) solution and trifluoromethanesulfonic acid trimethyl silyl ester (1.5mL, 7.75mmol) reaction.Stirring at room is after 1 hour under nitrogen atmosphere, is poured into this reaction soln in the ice-cold saturated sodium bicarbonate solution (20mL) and stirs 15 minutes.Water and this organic layer of salt water washing, and use dried over mgso.Removal of solvent under reduced pressure is also separated this resistates with 12.5% ethyl acetate/hexane by silica gel column chromatography, obtains anomer mixture 326 (1.25g, 62.9%), is soup compound.

6-chloro-9-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-L-glyceryl-penta-2-alkene furyl glycosyl] purine (326)

326:UV (MeOH) λ _Max265.0nm; Ultimate analysis (C ₁₆H ₂₂ClFN ₄O ₂Si) C, H, N.

6-chloro-2-fluoro-9-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-(α, β)-L-glyceryl-penta-2-alkene furyl glycosyl] purine (327 and 328)

Stirred 16 hours under the mixture room temperature with silylated 2-fluoro-6-chloropurine [by the 2-fluoro-6-chloropurine preparation of 1.170g (6.78mmol)] and anhydrous DCE (40mL).Be similar to 326 after treatment, obtain β anomer 327 (685mg, 1.70mmol, 30.0%) white foam and α anomer 328 (502mg by silica gel column chromatography (12% ethyl acetate/hexane) purifying, 1.25nunol, 22.1%) and faint yellow soup compound.

327：UV(MeOH)λ _max268.5nm。Ultimate analysis (C ₁₆H ₂₁F ₂ClN ₄O ₂Si) C, H, N., 328:UV (MeOH) λ _Max269.0nm.Ultimate analysis (C ₁₆H ₂₁F ₂ClN ₄O ₂Si) C, H, N.

6-chloro-9-(2,3-dideoxy-2-fluoro-(α, β)-L-glyceryl-penta-2-alkene furyl glycosyl) purine (329 and 330)

Anhydrous CH with 326 (1.2g, 3.12mmol) ₃CN (20mL) solution was processed and is stirred 1 hour with TBAF (the THF solution of 1M) (3.2ml, 3.2mmol).Behind the evaporating solvent, resistates is obtained β anomer 329 (296mg, 35%) white solid and α anomer 330 (380mg, 45%) foam by column chromatography (3% methyl alcohol/chloroform) purifying.

329：UV(MeOH)λ _max265.0nm.；330：UV(MeOH)λ _max265.0nm。

6-amino-2-fluoro-9-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl] purine (331) and

6-chloro-2-amino-9-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl] purine (332)

Ammonia with drying under the room temperature passes in anhydrous DME stirring, 327 (420mg, 1.04mmol) (35mL) solution, and spending the night passes into.Remove by filter salt and with this filtrate evaporated under reduced pressure.This resistates is obtained two kinds of compounds by silica gel column chromatography (25%EtOAc/ hexane) purifying, 331 (114mg, 0.30mmol) white solid and 332 (164mg, 0.41mmol) white solid.

331：UV(MeOH)λ _max268，5nm。Ultimate analysis (C ₁₆H ₂₃F ₂N ₅O ₂Si0.2 acetone) C, H, N; 332:UV (MeOH) λ _Max307.5nm.Ultimate analysis (C ₁₆H ₂₃FN ₅O ₂ClSi) C, H, N, CL.

6-amino-2-fluoro-9-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl] purine (333) and 6-chloro-2-amino-9-[5-O-(t-butyldimethylsilyl)-2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl] purine (334)

Ammonia with drying under the room temperature passes in dry DME stirring, 333 (420mg, 1.04mmol) (35mL) solution, and spending the night passes into.Remove by filter salt and with this filtrate evaporated under reduced pressure.This resistates is obtained two kinds of compounds by silica gel column chromatography (25%EtOAc/ hexane) purifying, 332 (150mg, 0.30mmol) white solid and 333 (150mg, 0.38mmol) white solid.

333：UV(MeOH)λ _max269.0nm。Ultimate analysis (C ₁₆H ₂₃F ₂N ₅O ₂Si0.3 acetone) C, H, N; 332:UV (MeOH) λ _Max309.5nm.Ultimate analysis (C ₁₆H ₂₃FClN ₅O ₂Si) C, H, N.

9-(2,3-dideoxy-2-fluoro-beta-glyceryl-penta-2-alkene furyl glycosyl) VITAMIN B4 (335)

With 329 (100mg, 0.369mmol) and saturated NH ₃/ MeOH (50mL) heated 24 hours at 90 ℃ in stainless steel cylinder.After being cooled to room temperature, removal of solvent under reduced pressure and with remaining soup compound by column chromatography 6%MeOH/CHCl ₃Obtain 335 (70mg, 75%) white solid as the eluent purifying.335：UV(H ₂O)λ _max258nm(ε18，800)(pH2)，258.5nm(ε18，800)(pH7)，258.5nm(ε19，100)(pH11)。Ultimate analysis (C ₁₀H ₁₀FN ₅O ₂.0.2H ₂O) C, H, N.

9-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) VITAMIN B4 (336)

With 330 (100mg, 0.369mmol) and saturated NH ₃/ MeOH (50mL) heated 24 hours at 90 ℃ in stainless steel cylinder.After being cooled to room temperature, removal of solvent under reduced pressure and with remaining soup compound by column chromatography 6%MeOH/CHCl ₃Obtain 335 (72mg, 78%) white solid as the eluent purifying.336：UV(H ₂O)λ _max258nm(ε21，100)(pH2)，259nm(ε21，500)(pH7)，29nm(ε22，600)(pH11)。Ultimate analysis (C ₁₀H ₁₀FN ₅O ₂.0.3MeOH) C, H, N.

9-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) xanthoglobulin (337)

With 329 (100mg, 0.369mmol), NaOMe (0.5M methanol solution) (2.94mL, 1.46mmol) and HSCH ₂CH ₂The mixture of OH (0.1mL, 1.46mmol) in methyl alcohol (20mL) refluxed 4 hours under nitrogen atmosphere.With this reaction mixture cooling, extremely do with Glacial acetic acid neutralization and vacuum-evaporation.This resistates is obtained 337 (74mg, 80%) white solid by silica gel column chromatography (10% methyl alcohol/chloroform) purifying.337：UV(H ₂O)λ _max247nm(ε12，400)(pH2)，247.5nm(ε13，000)(pH7)，253nm(ε13，100)(pH11)。Ultimate analysis (C ₁₀H ₉FN ₄O ₃.0.2H ₂O) C, H, N.

9-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) xanthoglobulin (338)

With 330 (100mg, 0.369mmol), NaOMe (0.5M methanol solution) (2.94mL, 1.46mmol) and HSCH ₂CH ₂The mixture of OH (0.1mL, 1.46mmol) in methyl alcohol (20mL) refluxed 4 hours under nitrogen atmosphere.With this reaction mixture cooling, extremely do with Glacial acetic acid neutralization and vacuum-evaporation.This resistates is obtained 338 (70mg, 80%) white solid by silica gel column chromatography (10% methyl alcohol/chloroform) purifying.338：UV(H ₂O)λ _max247.5nm(ε12，700)(pH2)，247.5nm(ε13，700)(pH7)，252.5nm(ε13，100)(pH11)。Ultimate analysis (C ₁₀H ₉FN ₄O ₃.0.3H ₂O) C, H, N.

2-fluoro-6-amino-9-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) purine (339)

Anhydrous acetonitrile (15mL) solution of 31 (101mg, 0.26mmol) was processed and stirred 30 minutes with TBAF (1 MTHF solution) (0.35mL, 0.35mmol).After the solvent evaporation, resistates is obtained 339 (64.7mg, 0.24mmol, 92.3%) white crystalline solid by column chromatography (9% methylene chloride/methanol) purifying.UV(H ₂O)λ _max269.0nm(pH7)。

2-fluoro-6-amino-9-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) purine (340)

Anhydrous acetonitrile (10mL) solution of 333 (73.4mg, 0.19mmol) was processed and stirred 30 minutes with TBAF (1 M THF solution) (0.25mL, 0.25mmol).After the solvent evaporation, resistates is obtained 340 (46.2mg, 0.17mmol, 90.3%) white crystalline solid by column chromatography (9% methylene chloride/methanol) purifying.UV(H ₂O)λ _max269.0nm(pH7)。

2-amino-6-chloro-9-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) purine (341)

Anhydrous acetonitrile (15mL) solution of 332 (143.5mg, 0.40mmol) was processed and stirred 30 minutes with TBAF (1 M THF solution) (0.60mL, 0.60mmol).After the solvent evaporation, resistates is obtained 341 (109mg, 0.382mmol, 90.3%) white crystalline solid by column chromatography (5% methylene chloride/methanol) purifying.UV(H ₂O)λ _max308.5nm(pH7)。

2-amino-6-chloro-9-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) purine (342)

Anhydrous acetonitrile (10mL) solution of 334 (145mg, 0.36mmol) was processed and stirred 30 minutes with TBAF (1 MTHF solution) (0.50mL, 0.50mmol).After the solvent evaporation, resistates is obtained 342 (99.9mg, 0.35mmol, 96.4%) white crystalline solid by column chromatography (9% methylene chloride/methanol) purifying.UV(H ₂O)λ _max309.0nm(pH7)。

9-(2,3-dideoxy-2-fluoro-beta-L-glyceryl-penta-2-alkene furyl glycosyl) guanine (343)

With 341 (63.6mg, 0.223mmol), 2 mercapto ethanol (0.06mL, 0.89mmol) and 1N NaOMe (0.89mL, 0.89mmol) refluxed 5 hours under nitrogen atmosphere in methyl alcohol (10mL).With the cooling of this mixture, with the Glacial acetic acid neutralization and be evaporated to dried.This resistates is obtained 343 (30.1mg, 0.113mmol, 50.7%) white solid by column chromatography (12% methylene chloride/methanol) purifying.UV(H ₂O)λ _max253.5nm(pH7)。

9-(2,3-dideoxy-2-fluoro-alpha-L-glyceryl-penta-2-alkene furyl glycosyl) guanine (344)

With 342 (59.3mg, 0.208mmol), 2 mercapto ethanol (0.07mL, 1.04mmol) and 1N NaOMe (1.04mL, 1.04mmol) refluxed 5 hours under nitrogen atmosphere in methyl alcohol (10mL).With the cooling of this mixture, with the Glacial acetic acid neutralization and be evaporated to dried.This resistates is obtained 344 (28.0mg, 0.105mmol, 50.5%) white solid by column chromatography (12.5% methylene chloride/methanol) purifying.UV(H ₂O)λ _max253.0nm(pH7)。

Synthetic suitable-(±)-carbocyclic ring d4 cytidine(C and 5 '-triphosphate thereof

With reference to scheme 11, by diallyl diethyl malonate (701) beginning, the yield with 78% synthesizes 4-ethoxycarbonyl-1, and 6-heptadiene (702) (W.A.Nugent, J.Am.Chem.Soc., 1995,117,8992-8998).Yield with 71% is by compound 702 synthetic compounds 703 (L.E.Martinez, J.Org.Chem., 1996,61,7963-7966) and with 43% yield by compound 704 synthetic compounds 705 (D.M.Hodgson, J.Chem.Soc.Perkin Trans.1,1994,3373-3378).Key intermediate is suitable-(±)-3-acetoxyl group-5-(acetoxy-methyl) cyclopentenes (708) or can from cyclopentadiene and formaldehyde acetic acid with the synthetic (E.A.Saville-Stones of Prins reaction, J.Chem.Soc.PerkinTrans.1,1991,2603-2604), although it has the low and inseparable problem of yield; Perhaps can be synthetic by bicyclic lactone, its through multi-step by the reaction of 4 steps synthetic (F.Burlina, Bioorg.Med.Chem.Lett., 1997,7,247-250).Although need the bicyclic lactone of synthesis of chiral, rear kind of method obtains chirality 708[(-)-enantiomer].By 5-flurocytosine and the synthetic N of acetic acid p-nitrophenyl ester ⁴-ethanoyl-5-flurocytosine (A.S.Steinfeld, J.Chem.Research (M), 1979,1437-1450).

Scheme 11

Experimental section

Introduction: all unless otherwise indicated reagent all directly uses.Anhydrous reagent is available from AldrichChemical Co..Measure fusing point (M.p.) and without calibration with electrothermal digit melting point apparatus. ¹H and ¹³C NMR spectrum at room temperature detects with Varian Unity Plus 400 spectrographs and with the ppm of internal standard substance tetramethylsilane downfield report.

4-ethoxycarbonyl-1,6-heptadiene (702)

With diallyl diethyl malonate (701; 50g, 208mmol), the mixture of sodium cyanide (20.7g, 422mmol) and DMSO (166mL) was 160 ℃ of heating 6 hours.After being cooled to room temperature, this mixture is joined in the 400mL water also with hexane (4 * 100mL) extraction products.After solvent removed under reduced pressure, (42-43 ℃/1Torr) obtain 702 of 27.34g (78%) is colourless liquid with this resistates distillation. ¹H?NMR(400MHz，CDCl ₃)65.80-5.70(m，2H，2CH＝CH ₂)，5.10-5.02(m，4H，2CH＝CH ₂)，4.14(q，2H，J＝7.2Hz，OCH ₂)，2.54-2.48(m，1H，CH)，2.41-2.34，2.29-2.23(2m，4H，2CH ₂)，1.25(t，J＝7.2Hz，3H，CH ₃)。

(±)-3-aleprolic acid ethyl ester (703)

In flame-dried 500mL flask, add 2,6-bromophenesic acid (1.20g, 4.76 mmol), tungsten oxychloride (0.813g, 2.38mmol) and dry toluene (25mL).Under nitrogen atmosphere with gained suspension reflux 1 hour, vacuum evaporating solvent then.Solid residue is smashed also vacuum-drying 30 minutes with scraper.In this resistates, add toluene (160mL), Et ₄Pb (1.54g, 4.76mL) and 702 (22g, 131.0mmol).Under nitrogen atmosphere, this mixture was heated 1.5 hours under 90 ℃.After being cooled to room temperature, this mixture is passed through diatomite filtration, and clean diatomite with t-butyl methyl ether.With 1% sodium hydroxide solution, water and salt water washing, and reduction vaporization concentrates with the filtrate that merges.(37-38 ℃/1Torr) obtain 703 of 13.06g (71%) is colourless liquid with this resistates distillation. ¹H NMR (400MHz, CDCl ₃) δ 5.67 (s, 2H, CH=CH), 4.14 (q, 2H, J=7.2Hz, OCH ₂), 3.11 (five small peaks, J=7.6Hz, 1H, CH), 2.65 (d, J=7.6Hz, 4H, 2CH2), 1.27 (t, J=7.2Hz, 3H, CH ₃).

(±)-1-(methylol)-3-cyclopentenes (704)

In cold (78 ℃) solution of anhydrous THF (150mL), add LiAlH to 703 (7g, 50mmol) ₄(1M THF solution, 25mL, 25mmol), and this reaction soln stirred under argon atmospher 4 hours under-78 ℃.Then allow this reaction soln be warming up to 0 ℃, and add successively 2.5mL water, 2.5mL 15% sodium hydroxide and 7.5mL water.After being warming up to room temperature, leaching precipitation and clean this diatomite with hot ethyl acetate by diatomite.The filtrate that merges is used 0.1N sodium hydroxide and salt water washing, and dry (sal epsom) filters, and concentrated and vacuum-drying obtains 704 of 4.294g (84%), is weak yellow liquid. ¹H?NMR(400MHz，CDCl ₃)δ5.68(s，2H，2CH＝CH)，

3.57(d，J＝6.0Hz，2H，CH ₂OH)，2.54-2.48(m，3H，CH+CH ₂)，2.15-2.10(m，2H，CH ₂)。

Suitable-(+)-4-(methylol)-1,2-cyclopentane epoxide (705)

To 704 (930mg; 9.1mmol) and anhydrous methylene chloride (20mL) solution of vanadyl Acetyl Acetone (10mg) in; drip tertiary butyl formic acid [3M dichloromethane solution; (70% (weight) is in water by tertiary butyl formic acid; 41mL, 0.3mol) and methylene dichloride (59mL) by dry (2 * MgSO ₄) and at 4 dust molecular sieves preservation preparation, 10mL, about 30mmol].Stir under the room temperature after 24 hours, (15% solution 60mL), and will stir 6 hours under this mixture room temperature to add sodium sulfite aqueous solution.Separate organic layer, with saturated sodium bicarbonate and salt water washing, and concentrated.This resistates is obtained 705 of 460mg (43%) at silica gel with hexane/ethyl acetate (2: 1) wash-out purifying by flash chromatography, be colourless liquid. ¹H?NMR(400MHz，CDCl ₃)δ3.54(s，2H，(CH) ₂O)，3.49(t，J＝4.0Hz，2H，CH ₂OH)，2.95(bs，1H，OH)，2.44-2.40(m，1H，CH)，2.05-2.02(m，4H，2?CH ₂)。 ¹³C?NMR(100MHz，CDCl ₃)δ66.9(d，(CH) ₂O)，59.2(t，CH ₂OH)，36.5(d，CH)，31.4(t，2?CH ₂)。

Suitable-(±)-3-acetoxyl group-5-(acetoxy-methyl) cyclopentenes (708)

In dehydrated alcohol (100mL) solution of phenylbenzene diselenide (2.70g, 8.65mmol), add sodium borohydride in batches.Stir this solution to yellow and become colorless, then add anhydrous THF (10mL) solution of 705 (1.70g, 14.4mmol).With this reaction soln reflux 1 hour under nitrogen atmosphere, then vacuum evaporating solvent.In this resistates, add ethyl acetate (80mL) and water (30mL).Separate organic layer, use the salt water washing, dry (sal epsom) filters, concentrated and vacuum-drying.With gained (±)-1-hydroxyl-4-(methylol)-2-(phenyl seleno)-pentamethylene (706; Faint yellow oily thing) the not purified the next step that is directly used in.In crude product 706, add anhydrous methylene chloride (60mL), triethylamine (30mL, 216mmol) and DMAP (50mg).Gained solution is cooled to 0 ℃, and drips acetic anhydride (14.7g, 144mmol).After the stirred overnight at room temperature, evaporating solvent obtains (±)-1-acetoxyl group-4-(acetoxy-methyl)-2-(phenyl seleno)-pentamethylene (707 under argon atmospher; Faint yellow oily thing).In 707 cold (0 ℃) methylene dichloride (50mL) solution that contains 3 pyridines, in 5 minutes, add 30% superoxol (20ml)., at room temperature stirred again 30 minutes after 30 minutes 0 ℃ of stirring, add methylene dichloride (50mL) and dilute this reaction mixture.Separate organic phase, water, saturated sodium bicarbonate and salt water washing, dry (sal epsom) filters and vacuum evaporation.This resistates is obtained 708 of 2.254g (79%, 3 step total recovery) at silica gel with the 0-10% eluent ethyl acetate purifying that is present in the hexane by flash chromatography, be light brown liquid. ¹H?NMR(400MHz，CDCl ₃)δ6.01-6.00，5.92-5.90(2m，2H，CH＝CH)，5.66-5.64(m，1H，H-3)，4.04(d，J＝6.8Hz，2H，CH ₂O)，2.98-2.92(m，1H，H-5)，2.53-2.46(m，1H，H-4a)，2.08，2.04(2s，6H，2CH ₃)，1.60-1.54?(m，2H，H-4b)。 ¹³C?NMR(100MHz，CDCl ₃)δ171.1，170.9(2s，2C＝O)，137.0，131.4(2d，CH＝CH)，79.2(d，C-3)，67.4(t，CH ₂O)，43.7(d，C-5)，33.4(t，C-4)，21.3，20.9(2q，2CH ₃)。

Suitable-(±)-carbocyclic ring-5 '-O-ethanoyl-2 ', 3 '-two dehydrogenations-2 ', 3 '-dideoxy-5-fluorine cytidine (709)

5-flurocytosine (258mg, 2mmol) and NaH (58mg, 2.4mmol) were heated 30 minutes at 70 ℃ in the oil bath of suspension in preheating in anhydrous DMSO (15mL).Then, gained solution is cooled to room temperature, and (the PPh that adds Pd respectively ₃) ₄Anhydrous THF (2mL) solution of (73mg, 0.063mmol) and 708 (298mg, 1.5mmol).This reaction mixture was stirred 3 days at 70 ℃ under argon atmospher.After the vacuum-evaporation desolventizing, this resistates is processed with methylene dichloride (50mL).Leach precipitation and clean this diatomite with methylene dichloride by diatomite.The filtrate that merges is concentrated, and this resistates is obtained 709 of 40mg (10%) at silica gel with the 0-5% methanol-eluted fractions purifying that is present in the methylene dichloride by flash chromatography, be the light brown solid.Obtain pure product with ethanol/methylene/hexane recrystallization, be white powder.M.p.182-184℃。 ¹H?NMR(400MHz，CDCl ₃)δ7.43(d，J＝6.0Hz，1H，H-6)，6.18-6.16(m，1H，H-3’)，5.83-5.81(m，1H，H-2’)，5.73-5.71(m，1H，H-1’)，4.23-4.21，4.08-4.04(2m，2H，CH ₂O)，3.14-3.12(m，1H，H-4’)，2.92-2.84(m，1H，.H-6’a)，2.08(s，3H，CH ₃)，1.41-1.35(m，1H，H-6’b)。

Suitable-(±)-carbocyclic ring-N ⁴, 5 '-O-diacetyl-2 ', 3 '-two dehydrogenations-2 ', 3 '-dideoxy-5-fluorine cytidine (710)

To be similar to 709 method, by 708 (560mg, 2.828mmol) and N ⁴-ethanoyl-5-flurocytosine (726mg, 4.24mmol) preparation title compound 710:560mg (64%, brown oil).With the not purified the next step that is directly used in of this crude product.

Suitable-(±)-carbocyclic ring-N ⁴, 5 '-O-diacetyl-2 ', 3 '-two dehydrogenations-2 ', 3 '-zalcitabine (711)

To be similar to 709 method, by 708 (272mg, 1.37mmol) and N ⁴The white powder of-ethanoyl cytosine(Cyt) (316mg, 2.06mmol) preparation title compound 711:108mg (27%).M.p.169.5-171.5℃。 ¹H?NMR(400MHz，CDCl ₃)δ8.80(bs，1H，NH)，7.72(d，J＝6.8Hz，1H，H-6)，7.39(d，J＝6.8Hz，1H，H-5)，6.19-6.17(m，1H.H-3’)，5.86-5.81(m，1H，H-2’)，5.77-5.75(m，1H，H-1’)，4.17-4.13，4.07-4.02(2m，2H，CH ₂O)，3.18-?3，10(m，1H，H-4’)，2.96-2.88(m，1H.H-6’a)，2.27，2.06(2s，6H，2?CH ₃)，1.43-1.37(m，1H，H-6’b)。 ¹³C?NMR(100MHz，CDCl ₃)δ170.8(s，2C＝O)，162.0(s，C-4)，155.6(s，C-2)，145.3(d，C-6)，139.2(d，C-3’)，130.0(d，C-2’)，96.8(d，C-5)，66.3(t，C-5’)，62.8(d，C-1’)，44.2(d，C-4′)，34.7(t，C-6’)，25.0，20.9(2q，2?CH ₃)。

Suitable-(±)-carbocyclic ring-2 ', 3 '-two dehydrogenations-2 ', 3 '-dideoxy-5-fluorine cytidine (712)

Add in the flask that 709 (33mg, 0.12mmol) are housed NaOMe (methanol solution of 0.5M, 0.5mL).With this reaction soln stirring at room 1 hour, and vacuum evaporating solvent.This resistates is obtained 712 of 17mg (61%) at silica gel with the 5-10% methanol-eluted fractions purifying that is present in the methylene dichloride by flash chromatography, be the light brown solid.Obtain pure products with ethanol/methylene/hexane recrystallization, be white powder.M.p.205.5-206.0℃. ¹HNMR(400MHz，DMSO-d ₆)δ7.66(d，J＝6.0Hz，1H，H-6)，7.60，7.40(2bs，2H，NH ₂)，6.06-6.05(m，1H，H-3’)，5.68-5.65(m，1H，H-2’)，5.53-5.50(m，1H，H-1’)，4.77-4.75(m，1H，H4’)，3.50-3.48，3.41-3.37(2m，2H，H-5’)，2.79-2.77(m，1H，H-6’a)，1.34-1.27(m，1H，H-6’b)。 ¹³C?NMR(100MHz，DMSO-d ₆)δ157.0(d，J _C-F＝11.9Hz，C-4)，154.0(3，C-2)，139.2(d，C-3’)，135.8(d，J _C-F＝241.3Hz，C-5)，130.2(d，C-2’)，126.8(d，J _C-F＝11.8Hz，C-6)，63.5(t，C-51)，61.3(d，C-1’)，47.2(d，C-4)，33.3(t，C-6’)。MS(FAB)m/e?226(MH ⁺)。Ultimate analysis (C ₁₀H ₁₂FN ₃O ₂) theoretical value C 53.33, H 5.37, and N 18.66; Measured value C53.10, H 5.40, and N 18.44.Be similar to aforesaid method, also prepared compound 712:320mg (59%, white powder) by 710 (750mg, 2.42mmol).

Suitable-(±)-carbocyclic ring-2 ', 3 '-two dehydrogenations-Zalcitabine (713)

To be similar to the method for preparation 712, by compound 711 (75mg, 0257mmol) preparation title compound 713:48mg (90%, white solid).M.p.200-201℃。 ¹H?NMR(400MHz，DMSO-d ₆)δ7.40(d，J＝7.2Hz，1H，H-6)，7，03，6.95(2bs，2H，NH ₂)，6.07-6.05(m，1H，H-3’)，5.67(d，J＝7.2Hz，1H，H5)，5.65-5.64(m，1H，H-2’)，5.55-5.52(m，1H，H-l’)，4.71-4.68(m，1H，H4’)，3.43-3.36(m，2H，H-5’)，2.78-2.76(m，1H，H-6’a)，1.24-1.18(m，1H，H-6’b). ¹³C?NMR(100MHz，DMSO-d ₆)δ165.5(s，C-4)，155.8(s，C-2)，142.2(d，C-6)，138.6(d，C-3’)，130.5(d，C-2’)，93.7(d，C5)，63.9(t，C-5’)，60.8(d，C-1’)，47.3?(d，C-4’)，34.0(t，C-6’)。MS(FAB)m/e?208(MH ⁺)。Ultimate analysis (C ₁₀H ₁₃N ₃O ₂) theoretical value D 57.96, H 6.32, and N 20.28; Measured value C 57.35, H 6.27, and N 20.02.HRMS (FAB) theoretical value (C ₁₀H ₁₄N ₃O ₂): 208.1086; Measured value 208.1088.

Suitable-(±)-carbocyclic ring-2 ', 3 '-two dehydrogenations-2 ', 3 '-dideoxy-5-fluorine CTP, triethyl hydrogen ammonium salt (714)

In the dry DMF (0.3mL) of 712 (10mg) and pyridine (0.1ml) solution, add 2-chloro-4H-1, the anhydrous Isosorbide-5-Nitrae of 1M of 3,2-benzo dioxa phosphorin-4-ketone-two _ alkane (0.05mL) solution.To stir 15 minutes under this reaction room temperature.Then, add successively tetra-sodium-Bu ₃1M dry DMF (0.12mL) solution and the Bu of N ₃N (0.05mL).Restir dripped I after 15 minutes under the room temperature in mentioned solution ₂/ H ₂The solution of O/ pyridine/THF is not until the color of iodine takes off (about 0.5mL), then with this mixture vacuum evaporation.This resistates is dissolved in the water (2mL), and (3 * 1mL) washings are filtered, and pass through FPLC purifying (post: HiLoad 26/10 Q Sepharose Fast Flow with methylene dichloride; Buffer A: 0.01M Et ₃NHCO ₃Buffer B: 0.7 M Et ₃NHCO ₃Flow velocity: 10mL/ minute; Gradient: 10% during with from the outset 0% to 4 minute of B was increased to 100% in the time of 64 minutes).Collect and suitable fraction freeze-drying is obtained 714, be colourless soup compound.HPLC[post: 100 * 4.6mm Rainin HydroporeSAX ion-exchange; Buffer A: 10mM NH ₄H ₂PO ₄At 10% methyl alcohol/H ₂Among the O (pH 5.5); Buffer B: 125mM NH ₄H ₂PO ₄At 10%MeOH/H ₂Among the O (pH5.5); Flow velocity: 1.0mL/min; Gradient: with B 0% 100% when increasing to 25 minutes from the outset], the residence time: 11.9 minutes.MS(FAB)m/e?464([M-H] ⁺)。

Suitable-(±)-carbocyclic ring-2 ', 3 '-two dehydrogenations-Zalcitabine 5 '-phosphoric acid salt (715)

To be similar to the method for preparation 714, by 713 preparation title compounds 715.HPLC (condition the is the same) residence time: 12.1 minutes.MS(FAB)m/e?446([M-H] ⁺)。

(±)-carboxyl-D4FC-triphosphate is to the restraining effect of HIV1-RT

With r (I) _n(dC) _12-18Homopolymer templa-primer (Pharmacia, Piscataway, NJ) and HIV-1 heterodimer p66/51 reversed transcriptive enzyme (RT, Biotechnology General, Rehoval, Israel).Standard reaction mixture (100 μ l) contains 100mM Tris-Hcl (pH8.0), 50 mM KCl, 2mM MgCl ₂, 0.05 unit/ml r (I) _n

_12-18, 5mMDTT, 100 μ g/ml bovine serum albumins and 1 μ M ³H-dCTP (2Ci/mmol).With 3TCTP (0.001-50 μ M) as positive control.Compound was hatched one hour with the 1 HIV-1 RT of unit in reaction mixture under 37 ℃.Adding isopyknic cold 10%TCA/0.05% trisodium phosphate stops this reaction and hatched under 4 ℃ 30 minutes.Manually gathering in the crops instrument (Meriden.CT) with Packard is collected in the nucleic acid that precipitates on the glass fibre filter paper.Measure the count per minute (cpm) that radio-labeled absorbs with Packard 9600 Direct Beta counters.

IV. HIV (human immunodeficiency virus)-resistant activity

In one embodiment, disclosed compound or its medicinal derivative or salt or the medicament that contains these compounds is used for prevention and treatment HIV infects and relative disease are such as illness, Kaposis sarcoma, thrombopenic purpura and the opportunistic infection of AIDS related syndromes (ARC), persistent generalized lymphadenopathy (PGL), AIDS related neural disease, anti-HIV antibody positive and the HIV positive.In addition, these compounds or preparation can preventive use to prevent or to postpone anti-HIV antibody or HIV antigen positive or contacted the deterioration of clinical disease of the individuality of HIV.

Nucleosides suppresses the ability of HIV can pass through the kinds of experiments technology for detection.One of technology that below describes in detail is the inhibition that detects virus replication in human peripheral monokaryon (PBM) cell that is infected by HIV-1 (strain LAV), use phytohemagglutinin (PHA) stimulation.Measure the amount of the virus that produces by the reversed transcriptive enzyme that detects encoding viral.The amount of the enzyme that produces is proportional with the amount of produce virus.

Antiviral and Cytotoxicity tests:

In human peripheral monokaryon (PBM) cell, measure as previously mentioned Anti-HIV-1 Active (Schinazi, the R.F. of these compounds; McMillan, A.; Cannon, D.; Mathis, R.; Lloyd, R.M.Jr.; Peck, A.; Sonimadossi, J.-P.; St.Clair, M.; Wilson, J.; Furman, P.A.; Painter, G.; Choi, W.-B.; Liotta, D.C.Antimicrob.Agents Chemother.1992,36,2423; Schinazi, R.F.; Sommadossi, J.-P.; Saalmann, V.; Cannon, D.; Xie, M.-Y.; Hart, G.; Smith, G.; Hahn, E.Antimicrob.Agents Chemother.1990,34,1061).The stock solution (20-40mM) of these compounds of preparation in the DMSO of sterilization, then dilution is required concentration in perfect dielectric.In water, prepare AZT (AZT) stock solution.Use original shape HIV-1 _LA1MOI cell with 0.01.The virus that will be obtained by cell conditioned medium liquid after infection the 6th day by the reversed transcriptive enzyme experiment with poly-(rA) _nFew (dT) _12-18As the templa-primer quantitative assay.The existence of DMSO is on the not impact of output of virus in dilute solution (＜0.1%).Can in people PBM, CEM and Vero cell, estimate the toxicity of these compounds.The half effective ways (Adv.Enzyme Regul.1984,22,27) of describing with Chou and Talalay obtain antiviral EC by concentration-effect curve ₅₀With cytotoxicity IC ₅₀

The PBM cell (10 of phytohemagglutinin stimulation hepatitis B and the seronegative health donors of HIV-1, that survived three days will be derived from ⁶Cell/ml) with HIV-1 (strain LAY) with about hundred times of concentration to every milliliter of infection of 50% TCID (TICD 50), and the antiviral compound of different concns exist or not in the presence of cultivate.

After infecting about 1 hour, the medium that will contain test compound (concentration is 2 times of ultimate density in medium) or not contain compound joins flask (5ml; Final volume 10ml).With AZT as positive control.

Allow cells contacting virus (about 2 * 10 ⁵Dpm/ml detects by the reversed transcriptive enzyme experiment) and place again the carbonic acid gas incubator.HI-1 (strain LAV) derives from Center for DiseaseControl, Atlanta, Georgia.Be used for to cultivate PBM cell, results virus and measure reverse transcriptase activity method be described in the (J.Immun.Meth.76 such as McDougal, 171-183,1985) and (J.Clin.Moth.25 such as Spira, 97-99,1987), different is, and amphotericin B is not included in the medium (sees Schinazi etc., Antimicrob.AgentsChemother, 32,1784-1787 (1988); Id., 34:1061-1067 (1990)).

At the 6th day, with cell with supernatant liquor is transferred to the 15ml test tube and at about 900g centrifugal 10 minutes.Shift out the 5ml supernatant liquor and by 40000rpm centrifugal (Beckman 70.1 Ti whizzers) 30 minutes that virus is concentrated.Process the viral throw out of dissolving to detect the concentration of reversed transcriptive enzyme.The result represents with the sampling supernatant liquor of dpm/ml.The virus in a small amount that is obtained by supernatant liquor (1ml) can also and detect the concentration of reversed transcriptive enzyme by centrifugal concentrating before dissolving.

Half is (EC effectively ₅₀) (Antimicrob.AgentsChemother, 30,491-498 (1986) detects concentration by the half effective ways.In brief, as from detect reversed transcriptive enzyme, measuring, the inhibition percentage of virus is mapped to the micro-molar concentration of compound.EC ₅₀The concentration of viral growth compound when suppressing 50%.

With the similar condition of above-mentioned antiviral experiment, medicine exist or not in the presence of, can cultivate the not infection people PBM cell (3.8 * 10 that mitogen stimulates ⁵Cell/ml).Calculate cell with hematimeter and with trypanblue exclusion method after 6 days, such as Schinazi etc., Antimicrobial Agents and Chemotherapy, 22 (3), 499 (1982) is described.IC ₅₀The concentration of compound when being normal cell growth suppressed 50%.

Table 7 provides the HIV (human immunodeficiency virus)-resistant activity data of selected compounds.Use this experiment, measured (±)-carbocyclic ring-D4FC-TP (EC of 2 ', 3 '-unsaturated-5-fluorine cytidine ₅₀Be 0.40 μ M, and the EC of (±)-carbocyclic ring-D4C-TP (2 ', 3 '-unsaturated cytidine) ₅₀Be 0.38 μ M.

V. resistance of hepatitis B is active

Can estimate the ability that active compound suppresses to suppress in the 2.2.15 cell culture (with the HepG2 cell of hepatitis virus particles conversion) the hepatitis virus growth according to the method for following detailed description.

In this culture systems, detect the general introduction of antivirus action and analysis HBV DNA and open (Korba and Milman, 1991, Antiviral Res, 15:217) is described in detail in detail.Antiviral evaluation experimental preferably independently carry out to cell by two stages.In brassboard porose with equal densities in the inoculation of identical time.

Because cell is interior and the heritable variation of extracellular HBV DNA, when only having 3.5 times (for the HBV virion DNA) or 3.0 times (for HBV dna replication dna intermediate) that suppresses greater than the mean level (ML) of HBV dna form in the untreated cell, just think to have significance,statistical (P＜0.05).Be used in the concentration (it is a constant with each cytometer in these experiments) of integrating HBV DNA in each cell DNA preparation and calculate the concentration of HBV dna form in the cell, to guarantee the equivalent of cell DNA between the sample that separates.

The representative value of extracellular HBV virion DNA is 50 to 150pg/ml substratum (on average about 76pg/ml) in untreated cell.HBV dna replication dna intermediate is 50 to 100 μ g/pg cell DNAs (on average about 74pg/ μ g cell DNAs) in the cell in untreated cell.In a word, with the inhibition of HBV virion DNA concentration in the cell, owing to treat with antiviral compound, thereby not remarkable to the inhibition of HBV DNA concentration in the cell, and very slow (Korba and Milman, 1991 occur to get, Antiviral Res., 15:217).

The mode that can carry out hybridization analysis in these experiments produces HBV DNA to 3 * 10 in the cell of interior each cell of HBV DNA to 2-3 chromosome copies of the cell that is equal to about 1.0pg and 1.0pg/ml ⁵Virion/ml.

Carry out oxicity analysis whether to estimate any antivirus action of observing owing to the resultant action to cell viability.Herein method therefor be detect the toluylene red dyestuff (standard and be widely used in comprise at multiple virus-host system detect the cell viability material among HSV and the HIV) absorption.Oxicity analysis carries out in the flat culture plate in 96 holes.The cell that is used for oxicity analysis uses the scheme identical with antiviral evaluation as described below to cultivate and process with test compound.4 concentration of each compound test, each concentration is carried out in three parts of cultures (hole A, B and C).Absorption measurement toxic level with the centering red.With the dyestuff of inherence at 510nm (A _Sin) absorption value be used for quantitative analysis.This numerical value is with average A in 9 independent cultures _SinValue accounts for the percentage that remains on the untreated cell on 96 orifice plates identical with test compound and represents.

VI. anti-hepatitis C is active

By suppressing the HCV polysaccharase, by other enzyme of need in the replication cycle suppressing or can demonstrate compound by other currently known methods to have anti-hepatitis C active.These active methods of many measure are disclosed.

WO97/12033, apply on September 27th, 1996, applied for by Emory University, Iisting C.Hagedom and A.Reinoldus are the contrivers, require U.S.S.N.60/004, the right of priority of 383 (applications in September nineteen ninety-five) has been described the HCV polysaccharase detection that can be used for estimating the compounds of this invention activity.This application and invention belong to separately TrianglePharmaceutical s, Inc., Durham, North Carolina.Another kind of HCV polysaccharase detection method is by reports such as Bartholomeusz, " hepatitis C virus (HCV) the RNA polymerase detection of carrying out with clone HCV Nonstructural Protein "; Antiviral Therapy 1996:1 (Supp4) 18-24.

VI. treat abnormal cell proliferation

In selectable embodiment, these compounds are used for treating abnormal cell proliferation.By conventional screening experiment (carrying out as being subsidized by National Cancer Institute), or can estimate the activity of this compound with any other known screens choosing method (for example WO96/07413 is described).

According to the method in following cem cell or other tumor cell line detection method, the easily degree of the antitumour activity of assessing compound.Cem cell is human lymphoma cell's (a kind of T-lymphoblastoid cell line can derive from ATCC, Rockville, MD).Compound provides Useful Information for being familiar with this compound to the activity of tumour to the toxicity of cem cell.This toxicity is with IC ₅₀(μ M) detects.IC ₅₀Refer to suppress the test compound concentration of 50% growth of tumour cell in the substratum.IC ₅₀Lower, just more tool is active as this compound of antineoplastic agent.In a word, 2 '-the fluoro nucleosides shown anti-tumor activity and can be used for the treatment of the abnormality proliferation of cell, if its concentration shows toxicity during less than 50 μ M in the tumor cell line of CEM or other vitality tanacity, this concentration is more preferably less than 10 μ M, and is first-selected less than 1 μ M.Drug solution comprises the cycloheximide as positive control, with 2 times to ultimate density, minutes three parts place 50 μ l growth mediums, and are allowed to condition under 37 ℃ balance in 5% carbonic acid gas incubator.In 50 μ l growth mediums, add the logarithmic phase cell to ultimate density 2.5 * 10 ³(CEM and SK-MEL-28), 5 * 10 ³(MMAN, MDA-MB-435s, SKMES-1, DU-145, LNCap), 1 * 10 ⁴(PC-3, MCF-7) cells/well is also being hatched 3 (DU-145, PC-3, MMAN) under 5% carbon dioxide atmosphere under 37 ℃, 4 (MCF-7, SK-MEL-28, CEM), or 5 (SK-MES-1, MDA-MB-435s, LNCap) days.The cell that control wells comprises independent medium (blank) and do not contain medicine adds medium.After vegetative period, Cell Titer 96 test kits of 15 μ l are detected dye solutions (Promega, Madison, WI) join in each hole and with this plate and in 5% carbonic acid gas incubator, hatching under 37 ℃.Promega Cell Titer 96 test kits are detected stop solution and join in each hole and in this incubator, hatched 4-8 hour.Read absorption value with Biotek Biokinetics plate readout instrument (Biotek, Winooski, VT) at 570nm, with only having the hole of medium to make blank.Relatively calculate the average inhibition percentage of growth with untreated control.Method by Chou and Talalay is calculated IC ₅₀, IC ₉₀, slope and r value.Chou T-C, Talalay P. quantitative analysis dose-effect relationship: the synergy of multi-medicament or enzyme inhibitors.Adv?Enzyme?Regul?1984，22：27-55。

Can use this active compound to treat especially abnormal cell proliferation, and cell hyperproliferation particularly.The example of abnormal cell proliferation includes but not limited to: innocent tumor, and it includes but not limited to papilloma, adenoma, firoma, chondroma, osteoma, lipoma, vascular tumor, lymphangioma, leiomyoma, rhabdomyoma, meningioma, neuroma, ganglioneuroma, mole, pheochromocytoma, schwannoma, fibroadenoma, teratoma, the cryptomere mole, granuloma (granuosatheca), brenner tumor, gynandroblastoma, hilar cell tumor, matter between sex cords, the propagation of Leydig's cell tumor and thyoma and the smooth muscle cell in vascular tissue's spot evolution; Malignant tumour (cancer) includes but not limited to cancer, comprises renal cell carcinoma, prostate cancer, bladder cancer and gland cancer, fibrosarcoma, chondrosarcoma, osteosarcoma, liposarcoma, angiosarcoma, lymphangiosarcoma, leiomyosarcoma, rhabdosarcoma, myelocytic leukemia, erythroleukemia, multiple myeloma, neurospongioma, meningioma, thyoma, cystosarcoma phylloides, nephroblastoma, teratoma choriocarcinoma, t cell lymphoma on the skin (CTCL), be primary in dermatoma (for example, basal cell tumor, the squamous cytoma of skin, melanoma and bowen's disease), mammary cancer and other are invaded the tumour of profit skin, and the premalignant and malignant disease of Kaposi's sarcoma and mucosal tissue comprises the oral cavity, bladder and recial disease; Preneoplastic damage, fungi mycosis, psoriasis, dermatomyositis, similar rheumatism sacroiliitis, virus (for example, wart, simple scar rash and pointed condyloma), molluscum contagiosum, the premalignant and malignant disease (uterine cervix, vagina and vulva) of female genital tract.These compounds can also be used for drainage.

In this embodiment, active compound or its pharmaceutical salts are with the administration for the treatment of significant quantity, to reduce the hyper-proliferative of target cell.This active compound can be modified into and comprise targeting moiety, it concentrates this compound at avtive spot.Targeting moiety can comprise in conjunction with the antibody of target cell surface protein or antibody fragment, includes but not limited to EGF-R ELISA (EGFR), c-Esb-2 series acceptor and the vascular endothelial growth factor (VEGF) of acceptor.

VII. pharmaceutical composition

By using active compound or its medicinal derivative or its salt that is present in the significant quantity in pharmaceutical carrier or the thinner to the patient, can treat the patient who suffers from any disease of trouble of describing herein.These active substances can pass through any suitable administration with the liquid or solid form, for example, and oral, non-enteron aisle, intravenously, intradermal, subcutaneous or topical.For all above-mentioned illnesss, the preferred dosage of this compound is about 1 to 50mg/kg, preferred 1 to 20mg/kg body weight every day, and more putting it briefly is 0.1 to about 100mg every kg body weight/sky.The significant quantity of medicinal derivative can according to will with the weight of parent nucleosides calculate.If this derivative itself has activity, significant quantity can be used the weight estimation of derivative as mentioned above, or by other approach assessment well known by persons skilled in the art.

This compound includes but not limited to that with the easily administration of any suitable dosage form units every unit dosage form contains 7 to 3000mg, preferred 70 to 1400mg active constituents.Oral dosage is that 50-1000mg generally is easily.

The use of active constituent should reach about 0.2 to 70pM in the ideal, the peak plasma concentrations of the active compound of preferred about 1.0 to 10 μ m.For example, 0.1 to 5% solution (optionally being present in the physiological saline) that can be by the intravenous injection active compound or the bolus injection by active compound reach this purpose.

The concentration of active compound depends on absorption, deactivation and drainage rate and other factor well known by persons skilled in the art of medicine in pharmaceutical composition.Notice that this dose value also can change with the seriousness of the illness that will alleviate.Will also be understood that for any specific object; should use according to the needs of this individuality and use or suggestion the people's of this composition professional judgment; come within a certain period of time specific dosage regimen to be adjusted, and the concentration range that provides herein just for example but not be intended to scope or the enforcement of the composition of requirement for restriction protection.This active constituent can once use, and maybe can be divided into some less dosage with variable timed interval administration.

The preferred modes of this active compound is oral.Oral compositions generally contains inert diluent or edible carrier.They can be encapsulated in gelatine capsule or be compressed to tablet.For the oral therapeutic administration purpose, this active compound can use with the vehicle blending and with the form of tablet, lozenge or capsule.Medicinal adhesive and/or auxiliary material also can be used as the part of this composition.

Tablet, pill, capsule, lozenge etc. can contain any following component or kin compound: tackiness agent such as Microcrystalline Cellulose, tragacanth gum or gelatin; Vehicle such as starch or lactose, disintegrating agent such as alginic acid, Primogel or W-Gum; Lubricant such as Magnesium Stearate or Sterotes; Glidant such as colloidal silica; Sweeting agent such as sucrose or asccharin; Or correctives such as mentha camphor, wintergreen oil or orange flavoring.When this element formulation was capsule, except above-mentioned substance, it can contain liquid vehicle such as fatty oil.In addition, can contain various other materials in the unit dosage form, it modifies the physical form of this element formulation, for example, and sugar, lac or other enteric reagent.

This compound can be used as a kind of composition administration of elixir, suspensoid, syrup, wafer, chewing-gum etc.Except active ingredient beyond the region of objective existence syrup can contain sucrose as sweeting agent and some sanitas, dyes and dyestuffs and correctives.

This compound or its medicinal derivative or salt can also be mixed with other active substance that does not damage required effect, or mixed with the material of augmenting required effect, and for example microbiotic, antimycotic, antiphlogiston or other antiviral drug comprise other nucleoside compound.Can contain following component for non-enteron aisle, intradermal, solution or suspensoid subcutaneous or topical: sterilization thinner such as water for injection, salt brine solution, fixing oil, polyoxyethylene glycol, glycerine, propylene glycol or other synthetic; Antiseptic-germicide such as benzylalcohol or methyl p-hydroxybenzoate; Antioxidant such as xitix or sodium bisulfite; Sequestrant such as ethylenediamine tetraacetic acid (EDTA); Buffer reagent such as acetate, citrate or phosphoric acid salt and osmotic pressure regulator such as sodium-chlor or dextrose.Parenteral formulation can be encapsulated in ampoule, disposable syringe or glass or the plastic multiple dose vials.

If intravenous administration, preferred vector are physiological saline or phosphate buffered saline (PBS) (PBS).

In preferred embodiments, active compound is prepared with the carrier that prevents its quick elimination in vivo with this compound of protection, such as controlled release preparation, comprises implants and microencapsulated delivery system.Can use biodegradable, biocompatible polymkeric substance, for example, ethylidene vinylacetate, poly-acid anhydrides, polyglycolic acid, collagen, poe and poly(lactic acid).The method for preparing these preparations is apparent to those skilled in the art.These materials also can be bought from Alza Corporation.

Liposome suspensoid (comprising the liposome with the monoclonal antibody target cells infected of antiviral antigen) also is preferred pharmaceutical carrier.They can prepare according to method known to those skilled in the art, for example, U.S. Patent No. 4,522,811 (its integral body is incorporated herein by reference) are described.For example; by suitable fat (one or more) (such as stearyl-phosphoryl thanomin, stearyl-PC, peanut acyl phospholipids acyl group choline and cholesterol) is dissolved in inorganic solvent; then with this solvent evaporation; stay the film of dry fat on the surface of this container, can prepare Liposomal formulation.The aqueous solution of active compound or its monophosphate, diphosphate and/or triphosphate derivative is introduced in this container.Then, rotate this container by hand to allow lipid material disengage and disperse the lipid agglomerate from wall of container, so form liposome turbid liquor.

Be described of the present invention with reference to preferred embodiment.Based on foregoing description of the present invention, variation of the present invention and modified forms are apparent to those skilled in the art.

Claims (10)

1. prepare the method for the acetic ester of end group isomery, may further comprise the steps:

A) make chirality non-carbohydrate sugar encircle the electrophilic source reactant of precursor (4S)-5-(tert-butyl diphenyl siloxy-)-penta-4-lactone and fluorine, form the fluoro lactone:

Wherein TBDPSO is the tertiary butyl-phenylbenzene siloxy-;

B) described fluoro lactone is reduced to the fluoro lactol; And

C) described fluoro lactol acetyl is turned to the acetic ester of end group isomery.

2. the method for claim 1, it also is included in electrophilic source reactant forward direction non-carbohydrate sugar ring precursor (4S)-5-(tert-butyl diphenyl siloxy-)-penta-4-lactone of non-carbohydrate sugar ring precursor (4S)-5-(tert-butyl diphenyl siloxy-)-penta-4-lactone of step in a) and fluorine and adds pair (trimethyl silyl) lithamides (LiHMDS).

3. the process of claim 1 wherein that the electrophilic source of described fluorine is N-fluoro-(two) benzenesulfonimide.

4. the process of claim 1 wherein step in a) chirality non-carbohydrate sugar ring precursor (4S)-5-(tert-butyl diphenyl siloxy-)-penta-4-lactone and the electrophilic source reactant of fluorine before be translated into enolate.

5. the method for claim 1, it comprises that further the acetic ester that makes the end group isomery and purine or pyrimidine bases react to provide β-L-or β-D-2 '-alpha-fluoro nucleosides.

6. the method for claim 5, wherein said base is a kind of purine bases.

7. the method for claim 4, it also comprises makes described enolate and C _1-10The alkyl halide reaction.

8. the process of claim 1 wherein that described fluoro lactone is reduced by diisobutyl alanate (DIBAL-H).

9. the method for claim 5, wherein said β-L-or β-D-2 '-alpha-fluoro nucleosides is β-L-2 '-alpha-fluoro nucleosides.

10. the method for claim 5, wherein said β-L-or β-D-2 '-alpha-fluoro nucleosides is β-D-2 '-alpha-fluoro nucleosides.