CN106883280B - A kind of prodrug, preparation method, medical composition and its use - Google Patents

A kind of prodrug, preparation method, medical composition and its use Download PDF

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Publication number
CN106883280B
CN106883280B CN201611158070.5A CN201611158070A CN106883280B CN 106883280 B CN106883280 B CN 106883280B CN 201611158070 A CN201611158070 A CN 201611158070A CN 106883280 B CN106883280 B CN 106883280B
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hydroxyl
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CN106883280A (en
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周星露
刘兴国
董晓武
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Guangdong HEC Pharmaceutical
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HANGZHOU HERTZ PHARMACEUTICAL Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A61K31/7072—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/57—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C323/58—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/57—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C323/58—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton
    • C07C323/59—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups with amino groups bound to the carbon skeleton with acylated amino groups bound to the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/60—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton with the carbon atom of at least one of the carboxyl groups bound to nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention discloses a kind of prodrugs containing class nucleotide structure or its tautomer, stereoisomer shown in formula I, stereoisomer mixture or its pharmaceutically acceptable solvate, preparation method is also disclosed, and the pharmaceutical composition being made from it and its purposes as anti-hepatitis C drug.Such compound or its pharmaceutical composition can be used for the treatment of hepatitis C, while having the effects that protect hepatic tissue, liver cell, improve liver function, reduce aminopherase.

Description

A kind of prodrug, preparation method, medical composition and its use
Technical field
The invention belongs to field of medicinal chemistry.Specifically, the present invention relates to a kind of following general formula I compound represented or Its tautomer, stereoisomer, stereoisomer mixture, prodrug, pharmaceutically acceptable salt or its solvate, Preparation method, pharmaceutical composition and its purposes as anti-hepatitis C drug.
Background technique
Hepatitis C is a kind of serious prestige caused by Hepatitis C Virus (hepatitis C virus, hereinafter referred to as HCV) Coerce the liver diseases of human health.1978, hepatitis C virus was found for the first time；1989, complete gene sequencing, HCV quilt It is confirmed as non-A type, another main pathogens of non-hepatitis B.According to HCV genome heterogeneity characteristic, can divide at present For 6 seed types, 30 hypotypes.The type of infection has stronger region, and China is based on 1 type.2011 studies have shown that in In state HCV infection person, 1 type accounts for 58.2%, and wherein gene 1a type is 1.4%, genotype 1 b 56.8%.Currently, global range Interior HCV infection person accounts for the 3.3% of total world population more than 200,000,000.There are about 3,200,000 the infecteds, Chinese HCV patients in the U.S. is more than It 40000000 person-times, occupies first of the world, many HCV infection persons are hepatitis B even AIDS patient simultaneously, and which increase answering for treatment Polygamy.There is presently no the hepatitis C vaccines formally to get the Green Light, so preventing hepatitis propagation from still facing the challenge.Existing chronic sense Dye person's state of an illness is developing, it is contemplated that the following 10-20 will welcome onset peak.The standard scheme of clinical treatment hepatitis is Peg-IFN alpha-2b α combines Ribavirin, and criterion of cure is RNA of the blood testing less than HCV in 24 weeks after the treatment.It is this Therapeutic modality side effect is larger, including depressed, tired, influenza-like symptom and anemia etc.；Treatment cost is higher, and standard course for the treatment of needs 48 weeks are wanted, expense about 40,000 dollar.To solve the above problems, being directed to the anti-hepatitis of Hepatitis C virus RNA gene order in recent years The small molecule compound medicament research and development of virus rapidly becomes hot spot, and antiviral drugs will enter small-molecule drug from the interferon epoch Epoch.Currently, be widely studied there are many HCV protease inhibitor, wherein Telaprevir (tirrevir, VX-950, Trade name Incivek) and Boceprevir (Bo Xipuwei, SCH-503034, trade name Victrelis) criticized in 2011 years Quasi- listing.The inhibitor of non-structural protein NS5A is also a kind of specificity antivirus drug for acting on Hepatitis C virus RNA chain, more Kind HCV genotype virus all has significant inhibiting effect.In addition, using NS5B polymerase as the drug of target spot be divided into ucleosides and Non- two class of nucleosides polymerase inhibitors.Sofosbuvir therein is anti-HCV medicament the most efficient so far, can be effectively right HCV-Ab IgG genotype 1,2,3,4 and 6 infects.
Summary of the invention
The purpose of the present invention is to provide a kind of new HCV-Ab IgG virus and liver protection, the difunctional drug of protect liver, play it double The effect of function medicament has oral administration biaavailability high, and the good advantage of metabolisming property can be used for treating virus hepatitis etc. Disease.
It is an object of the present invention to provide prodrug shown in a kind of general formula I or its stereoisomer, stereoisomer is mixed Close object or its pharmaceutically acceptable solvate.
It is a further object to provide the preparation methods of compound provided by the invention.
A further object of the present invention is to provide prodrug shown in general formula I or its stereoisomer, stereoisomer mixing Object or its pharmaceutically acceptable salt or solvate protect liver cell, liver organization, improve liver function as NS5B inhibitor The purposes of energy, and the application in preparation treatment virus hepatitis.
It is also another object of the present invention to provide include prodrug shown in general formula I or its stereoisomer, stereoisomer One of mixture or its pharmaceutically acceptable solvate or a variety of pharmaceutical compositions.
Virus hepatitis is treated it is also another object of the present invention to provide a kind of, while protecting liver cell, liver organization, is changed The method of kind liver function.
The present invention uses technical solution as described below:
According to an aspect of the present invention, the present invention protects a kind of compound, with structure shown in general formula I or its Stereoisomer, stereoisomer mixture or its pharmaceutically acceptable salt or solvate, or have shown in general formula I Prodrug:
Wherein, X is hydroxyl or F, Cl, Br；R isLinker is Or be not present, A from the drug listed for treating or assisting in the treatment of hepatopathy or Its derivative；R1For H or C1-C5Alkyl；N is the integer of 1-19.
In the general formula I, further preferred compound be general formula II-A shown in the prodrug containing class nucleotide structure or Its stereoisomer, stereoisomer mixture or its pharmaceutically acceptable salt or solvate:
Wherein, X is hydroxyl or F, Cl, Br；Linker isOr it is not present；R1For H or C1-C5Alkyl；N is 1,2,3,4 Or 5；R3、R4It is each independently H or C1-C5Alkyl.
In the general formula II-A, further preferred compound be general formula II-Aa or Formula II-Ab shown in structure or Its tautomer, optical isomer, stereoisomer, stereoisomer mixture, prodrug, pharmaceutically acceptable salt or molten Object is closed in agent:
Wherein, X is hydroxyl or F, Cl, Br；Linker isOr it is not present；R1For H or C1-C5Alkyl；N is 1,2,3,4 Or 5；R3、R4It is each independently H or C1-C5Alkyl.
In the general formula I, compound further preferably be structure shown in general formula II-B or its tautomer, Optical isomer, stereoisomer, stereoisomer mixture, prodrug, pharmaceutically acceptable salt or solvate:
Wherein, X is hydroxyl or F, Cl, Br；Linker isOr it is not present；R1For H or C1-C5Alkyl；N is 1,2,3,4 Or 5；R3、R4It is each independently H or C1-C5Alkyl.
In the general formula II-B, structure or its optical siomerism shown in further preferably general formula II-Ba or Formula II-Bb Body, pharmaceutically acceptable salt or solvate:
Wherein, X is hydroxyl or F, Cl, Br；Linker isOr it is not present；R1For H or C1-C5Alkyl；N is 1,2,3,4 Or 5；R3、R4It is each independently H or C1-C5Alkyl.
In the general formula I, the prodrug containing Tiopronin structure shown in further preferably general formula II-A-A (1) or its Stereoisomer, stereoisomer mixture or its pharmaceutically acceptable solvate:
Wherein, X is hydroxyl or F, Cl, Br, and Linker isR1For H or C1-C5Alkyl, n are the integer of 1-19.
Alternatively, preferably, further preferably the following general formula II-A-A (2) is shown in the general formula II-A-A (1) Prodrug containing Tiopronin structure or its stereoisomer, stereoisomer mixture or its pharmaceutically acceptable solvent close Object:
In above formula: X, Linker A, R1, n be defined as above Formula II-A-A (1).
In the general formula II-A-A (1), the further preferably following prodrug containing Tiopronin structure or its solid Isomers, stereoisomer mixture or its pharmaceutically acceptable solvate:
Wherein, X is hydroxyl or F, Cl, Br；R1For H or C1-C5Alkyl, n 1,2,3,4 or 5.
Alternatively, preferably, further preferably containing Tiopronin structure below in the general formula II-A-A (1) Prodrug or its stereoisomer, stereoisomer mixture or its pharmaceutically acceptable solvate:
Wherein, X, R1, n is defined as above.
Alternatively, preferably, further preferably containing Tiopronin structure below in the general formula II-A-A (1) Prodrug or its stereoisomer, stereoisomer mixture or its pharmaceutically acceptable solvate:
Wherein, X, R1, n is defined as above.
In the general formula II-A-A (1), the further preferably following prodrug containing Tiopronin structure or its solid Isomers, stereoisomer mixture or its pharmaceutically acceptable solvate:
Wherein, R1For H or C1-C5Alkyl, n 1,2,3,4 or 5.
In the general formula I compound represented, further preferred particular compound is one of following compounds:
Or the stereoisomer of above-mentioned preferred compound, stereoisomer mixture or its pharmaceutically acceptable salt or molten Object is closed in agent.
It is a further object to provide the preparation methods of general formula II-A compound, which comprises 1) formula III Compound reacts to obtain with formula IV-A compound Formula V-A compound, 2) Formula V-A compound obtains Formula II-A by deprotection reaction Compound；
Wherein, X is hydroxyl or halogen (F, Cl, Br)；Linker is Or it is not present；N is 1,2,3,4 or 5；R1For H or C1-C5Alkyl；R3、R4It is each independently H or C1-C5Alkane Base；LG is leaving group, preferably halogen；Z is thiol protecting group, preferably trityl, 4- Methoxytrityl, 4, 4 '-dual-methoxy trityls.
It is a further object to provide the preparation methods of formula Ii-B compound, which comprises 1) formula III Compound reacts to obtain Formula V-B compound with formula IV-B compound；2) Formula V-B compound obtains Formula II-Bization through deprotection reaction Close object；
Wherein, X hydroxyl or halogen (F, Cl, Br)；Linker is Or it is not present；N is 1,2,3,4 or 5；R1For H or C1-C5Alkyl；R3、R4It is each independently H or C1-C5Alkane Base；LG is leaving group, preferably halogen；Z is thiol protecting group, preferably trityl, 4- Methoxytrityl, 4, 4 '-dual-methoxy trityls.
Further, a kind of preparation method of general formula II-A-Aa compound represented, can be by compound of formula III General formula V-A compound is obtained by alkylation reaction with compound of Formula IV, is then obtained by deprotection reaction.
Wherein, X F, Cl or Br；R1For H or C1-C5Alkyl；Y is leaving group, and Z is thiol protecting group.
A kind of preparation method of Formula II-A-Ab compound represented, can be by compound of formula III and general formula IV-B chemical combination Object obtains general formula V-B compound by alkylation reaction, then obtains by deprotection reaction:
Wherein, X is hydroxyl or halogen (F, Cl, Br)；R1For H or C1-C5Alkyl；N is the integer of 1-19；Y is leaving group Group；Z is thiol protecting group.
It is a further object to provide the preparation methods of general formula II-A-B compound represented, can be by III institute The compound shown is reacted through overprotection, general formula VII-1 compound represented is then obtained with aldehyde reaction, then and shown in general formula Esterification occurs for VIII-1A compound represented or acylation reaction obtains general formula IX-A-B compound represented, then sloughs protection Base obtains general formula II-1A-B compound represented.The active site of the reaction of the key intermediate of this preparation method is suitably protected, Facilitate the generation of reduction side reaction, reaction selectivity is high, and obtained intermediate, final product is with high purity, is easy to purify.And this The reaction that method is related to has the advantage that easy to operate, purifying is convenient and process controllability is good, is suitble to industrialization production.
Wherein, X is hydroxyl or halogen (F, Cl, Br)；R1For H or C1-C5Alkyl；R3、R4It is each independently H or C1-C5 Alkyl；P1For hydroxy-protective group, preferably trimethyl silicon substrate, triethyl group silicon substrate, t-Butyldimethylsilyl, triphenyl silicon Base；P2For amide NH blocking group, preferably tertbutyloxycarbonyl, benzyloxycarbonyl group, 4- methbxybenzyl-oxycarbonyl；P3It is sulfhydryl protected Group, preferably trityl, 4- Methoxytrityl, 4,4 '-dual-methoxy trityls.
It is a further object to provide the preparation methods of general formula II-B-B compound represented, can be by general formula Then VII-1 compound represented occurs esterification with general formula VIII-1B compound represented or acylation reaction obtains general formula IX- Then B-B compound represented sloughs blocking group and obtains general formula II-B-B compound represented.In the key of this preparation method The active site of the reaction of mesosome is suitably protected, and facilitates the generation for reducing side reaction, reaction selectivity is high, obtained centre Body, final product purity is high, are easy to purify.And the reaction that this method is related to has easy to operate, purifying convenience and technique can The good advantage of control property, is suitble to industrialization production.
Wherein, X, R1, R3, R4Definition with its definition in general formula II-B, P1For hydroxy-protective group, preferably three Methylsilyl, triethyl group silicon substrate, t-Butyldimethylsilyl, triphenyl silicon substrate；P2For amide NH blocking group, preferably tertiary fourth Oxygen carbonyl, benzyloxycarbonyl group, 4- methbxybenzyl-oxycarbonyl；P3For thiol protecting group, preferably trityl, 4- methoxyl group triphen Methyl, 4,4 '-dual-methoxy trityls.
It is a further object of the present invention to provide intermediate shown in following VII and its it is used to prepare anti-hepatitis C virus drug Purposes:
Wherein, X is hydroxyl or halogen (F, Cl, Br)；R1For H or C1-C5Alkyl；P1For hydroxy-protective group, preferably three Methylsilyl, triethyl group silicon substrate, t-Butyldimethylsilyl, triphenyl silicon substrate.
It is a further object of the present invention to provide intermediate shown in following VIII-A or its stereoisomer or its alloisomerisms Body mixture:
Wherein, P2For NH blocking group, preferably tertbutyloxycarbonyl, benzyloxycarbonyl group, 4- methbxybenzyl-oxycarbonyl；P3For mercapto Base base blocking group, preferably trityl, 4- Methoxytrityl, 4,4 '-dual-methoxy trityls etc..
It is a further object of the present invention to provide prodrug shown in following IX-A-B or its stereoisomer, stereoisomer is mixed Close object:
Wherein, X is hydroxyl or halogen (F, Cl, Br)；R1For H or C1-C5Alkyl；P1For hydroxy-protective group, preferably three Methylsilyl, triethyl group silicon substrate, t-Butyldimethylsilyl, triphenyl silicon substrate；P2For NH blocking group, preferably tertiary butyloxycarbonyl Base, benzyloxycarbonyl group, 4- methbxybenzyl-oxycarbonyl etc.；P3For thiol protecting group, preferably trityl, 4- methoxyl group triphen first Base, 4,4 '-dual-methoxy trityls etc..
It is a further object of the present invention to provide intermediate shown in following VIII-B or its stereoisomer or its alloisomerisms Body mixture:
Wherein, R3、R4It is each independently H or C1-C5Alkyl；P2For amide NH blocking group, preferably tertbutyloxycarbonyl, Benzyloxycarbonyl group, 4- methbxybenzyl-oxycarbonyl；P3For thiol protecting group, preferably trityl, 4- Methoxytrityl, 4, 4 '-dual-methoxy trityls.
It is a further object of the present invention to provide prodrug shown in following IX-B-B or its stereoisomer, stereoisomer is mixed Close object:
Wherein, X is hydroxyl or halogen (F, Cl, Br)；R1For H or C1-C5Alkyl；R3、R4It is each independently H or C1-C5 Alkyl；P1For hydroxy-protective group, preferably trimethyl silicon substrate, triethyl group silicon substrate, t-Butyldimethylsilyl, triphenyl silicon Base；P2For amide NH blocking group, preferably tertbutyloxycarbonyl, benzyloxycarbonyl group, 4- methbxybenzyl-oxycarbonyl；P3It is sulfhydryl protected Group, preferably trityl, 4- Methoxytrityl, 4,4 '-dual-methoxy trityls.
General formula I compound represented can be containing one or more chiral centres, and because stereoisomer may be present, i.e. mapping is different Or mixtures thereof structure body, diastereoisomer.Therefore, formula I- compound represented can be single isomers or different The mixture of structure body.
The present invention includes any prodrug forms of general formula I compound represented.
The invention also includes the pharmaceutical acceptable solvates of compounds of formula I.
The present invention also includes the pharmaceutically acceptable oxide and its officinal salt and acceptable solvent of general formula I compound represented Compound.
The invention also includes a variety of crystal forms of general formula I compound represented.
According to another aspect of the invention, the present invention provides general formula I compound represented or its tautomers, solid Isomers, the purposes of stereoisomer mixture, prodrug, pharmaceutically acceptable salt or its solvate press down as NS5B Preparation protects protection liver cell, liver organization simultaneously, improves the purposes of liver function, and in preparation for treating virus hepatitis etc. Purposes in the drug of disease.
In accordance with a further aspect of the present invention, change shown in the present invention also provides a kind of general formula I comprising therapeutically effective amount Close object or its tautomer, stereoisomer, stereoisomer mixture, prodrug, pharmaceutically acceptable salt or its solvent One of object or a variety of pharmaceutical compositions are closed, NS5B inhibitor is can be used as and the composition can optionally include Pharmaceutically acceptable carrier or excipient.
According to another aspect of the present invention, the present invention also provides a kind of NS5B inhibitor, logical containing therapeutically effective amount Formulas I compound represented or its tautomer, stereoisomer, it is stereoisomer mixture, prodrug, pharmaceutically acceptable One of salt or its solvate or a variety of and inhibitor can optionally include pharmaceutically acceptable carrier or figuration Agent.
The composition by therapeutically effective amount one or more general formula I compounds represented or its tautomer, solid Isomers, stereoisomer mixture, prodrug, pharmaceutically acceptable salt or its medicine solvate and at least one are pharmaceutically acceptable auxiliary Material composition.The selection of pharmaceutic adjuvant is different because of administration method and action character, usually filler, diluent, adhesive, wetting Agent, disintegrating agent, lubricant, emulsifier, suspending agent etc..Compound of formula I, its stereoisomer, stereoisomer mixture or its Shared ratio of the pharmaceutically acceptable solvate in above-mentioned composition is the 0.1%~99.9% of total weight, preferably 1%~99%.
The pharmaceutically acceptable carrier refers to the pharmaceutical carrier of pharmaceutical field routine, such as: diluent, such as water； Filler, such as starch, sucrose；Adhesive, such as cellulose derivative, alginates, gelatin, polyvinylpyrrolidone；Wetting agent, Such as glycerol；Disintegrating agent, such as agar, calcium carbonate and sodium bicarbonate；Sorbefacient, such as quaternary ammonium compound；Surfactant, such as ten Six alkanols；Absorption carrier, such as kaolin and soap clay；Lubricant, such as talcum powder, calcium stearate and magnesium stearate, He Juyi bis- Alcohol etc..Furthermore it is also possible to other adjuvants be added in described pharmaceutical composition, such as flavouring agent and sweetener.
The present invention also provides general formula I compound represented or its tautomer, stereoisomer, stereoisomer is mixed Close the preparation method of the pharmaceutical composition of object, prodrug, pharmaceutically acceptable salt or its solvate.Usually by general formula I Shown in the compound containing Tiopronin structure or its tautomer, stereoisomer, it is stereoisomer mixture, preceding Medicine, pharmaceutically acceptable salt or its solvate and pharmaceutically acceptable auxiliaries mix, and are made through conventional preparation method suitable for one Determine the form (dosage form) of approach application.Dosage form includes tablet, capsule, granule, pill, solution, suspension, emulsion, soft Cream, film, creme, aerosol, injection, suppository etc..Preferred tablet and capsule.
The dosage of the compounds of this invention is generally daily 1~1000mg, point single or multiple uses.But in necessity When, it can suitably deviate above-mentioned dosage.Professional can determine optimal dose as the case may be and professional knowledge.These situations Including the severity of disease, the individual difference of patient, the characteristic of preparation and administration route etc..
In addition, the present invention also provides general formula I compound represented or its tautomer, stereoisomer, it is three-dimensional different Structure body mixture, prodrug, pharmaceutically acceptable salt or its solvate or its pharmaceutical composition are as human medicine Purposes.
According to another aspect of the invention, the present invention also provides the method for the diseases such as treatment virus hepatitis, the methods General formula I compound represented or its tautomer, stereoisomer including applying therapeutically effective amount, stereoisomer mixing One of object, prodrug, pharmaceutically acceptable salt or its solvate or a variety of or of the invention described pharmaceutical compositions To patient.
Compound or composition provided by the invention can take orally, inject (vein, muscle, in subcutaneous and coronary artery), Sublingual, buccal, per rectum, per urethra, Via vagina, intranasal, sucking or topic route application.Preferred approach is oral.For When oral, it can be made into conventional solid pharmaceutical preparation, such as tablet, pulvis, granula, capsule, or liquid preparation is made, such as water Or oil-suspending agent or other liquid preparations, such as syrup；When for parenteral administration, can be made into the solution of injection, water or Oleaginous suspension etc..
The present invention also provides general formula I compound represented or its tautomer, stereoisomer, stereoisomer is mixed Object, prodrug, pharmaceutically acceptable salt or its pharmaceutically acceptable solvate are closed, in the people's medication for preparing NS5B inhibitor Purposes in object.
The present invention also provides general formula I compound represented or its tautomer, stereoisomer, stereoisomer is mixed Close object, prodrug, pharmaceutically acceptable salt or its solvate further drug combination with other treatment hepatopathy.It is at least one Selected from following treatment: interferon, interferon beta, interferon gamma and interferon ω；Interleukin, including IL-10 and interleukin 12；Ribavirin；Interferon-' alpha ' or glycol interferon alpha are used in combination with Ribavirin or Levovirin；Left-handed Wei Woods；Protease inhibitors, including NS3 inhibitor, NS3/4A inhibitor；NS5A inhibitor；Helicase inhibitors；Polymerase inhibits Agent, including HCV RNA polymerase and NS5B polymerase inhibitors；Gliotoxin；IRES inhibitor；Antisense oligonucleotides；Thiazole Alkane derivatives；N- benzanilide, ribozyme；Another nucleosides, nucleoside prodrugs or nucleoside derivates；1- amino-alkylcyclohexane； Antioxidant, including vitamin E；Squalene；Amantadine；Bile acid；N- (phosphonoacetyl)-L-Aspartic acid；Benzene dicarboxyl Amide；Polyadenylic acid；Benzimidazole；Thymosin extrasin；Prevent vaccine；Immunomodulator, a kind of IMPDH inhibitor；Silymarin；Water Fly silibin-phosphatidyl choline endosome；And Mycophenolate Mofetil.
" pharmaceutically acceptable " refers to compounds some in this way, raw material, composition and/or dosage form, they are cured rationally Learn judgement in the range of, be suitable for contacted with patient tissue and without excessive toxicity, irritation, allergy or with reasonable benefit The symmetrical other problems of benefit/Hazard ratio and complication, and effective for given application.
Term " halogen " and " halogenated " are used interchangeably in the present invention, refer to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I)。
Terminology used in the present invention " alkyl " or " alkyl group ", indicate contain 1-20 carbon atom, the straight chain of saturation or Branch univalent hydrocarbyl group, wherein the substituent group institute that the alkyl group can be described optionally by one or more present invention Replace.In one embodiment, alkyl group contains 1-6 carbon atom；In another embodiment, alkyl group contains 1-4 A carbon atom；Also in one embodiment, alkyl group contains 1-3 carbon atom.The example of alkyl group includes, but and unlimited In methyl (Me ,-CH3), ethyl (Et ,-CH2CH3), n-propyl (n-Pr ,-CH2CH2CH3), isopropyl (i-Pr ,-CH (CH3)2), normal-butyl (n-Bu ,-CH2CH2CH2CH3), isobutyl group (i-Bu ,-CH2CH(CH3)2), sec-butyl (s-Bu ,-CH (CH3)CH2CH3), tert-butyl (t-Bu ,-C (CH3)3), etc..
" pharmaceutically acceptable salt " used in the present invention refers to the organic salt and inorganic salts of the compound of the present invention.Medicine Acceptable salt is known to us in fields on, such as document: S.M.Berge et al., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977,66:1-19. documented.The salt that pharmaceutically acceptable nontoxic acid is formed includes, but is not limited to, with amino base The inorganic acid salt that group's reaction is formed has hydrochloride, hydrobromate, phosphate, sulfate, perchlorate and acylate such as acetic acid Salt, oxalates, maleate, tartrate, citrate, succinate, malonate, or by recorded in books, literature Other methods such as ion-exchanges obtain these salt.Other pharmaceutically acceptable salts include adipate, and alginates resist Bad hematic acid salt, aspartate, benzene sulfonate, benzoate, bisulphate, borate, butyrate, camphor hydrochlorate, camphor sulphur Hydrochlorate, cyclopentyl propionate, digluconate, lauryl sulfate, esilate, formates, fumarate, Portugal Heptose hydrochlorate, glycerophosphate, gluconate, Hemisulphate, enanthate, caproate, hydriodate, 2- hydroxy-ethanesulfonic acid Salt, lactobionate, lactate, laruate, lauryl sulfate, malate, mesylate, 2- naphthalene sulfonate, niacin Salt, nitrate, oleate, palmitate, pamoate, pectate, persulfate, 3- phenylpropionic acid salt, picrate, spy penta Hydrochlorate, propionate, stearate, rhodanate, tosilate, undecylate, valerate, etc..Pass through alkali appropriate Obtained salt includes alkali metal, alkaline-earth metal, ammonium and N+(C1-4Alkyl)4Salt.The present invention is also intended to contemplate any included N's The compound of group is formed by quaternary ammonium salt.Water-soluble or oil-soluble or dispersion product can be obtained by quaternization.Alkali Metal or alkali salt include sodium, lithium, potassium, calcium, magnesium, etc..Pharmaceutically acceptable salt further comprises appropriate, nontoxic Ammonium, the amine cation that quaternary ammonium salt and gegenions are formed, such as halide, hydroxide, carboxylate, hydrosulphate, phosphoric acid Compound, nitric acid compound, C1-C8Sulphonic acid compound and aromatic sulphonic acid compound.
" solvate " of the invention refers to that one or more solvent molecules and the compound of the present invention are formed by association Object.The solvent for forming solvate includes, but is not limited to, water, isopropanol, ethyl alcohol, methanol, dimethyl sulfoxide, ethyl acetate, second Or mixtures thereof acid, ethanol amine.Term " hydrate " refers to that solvent molecule is that water is formed by associated matter.
When the solvent is water, term " hydrate " can be used.In one embodiment, a compounds of this invention Molecule can be combined with a hydrone, such as monohydrate；In another embodiment, a compounds of this invention molecule It can be combined with more than one hydrone, such as dihydrate, in yet another embodiment, a compounds of this invention point Son can be combined with the hydrone less than one, such as semihydrate.It should be noted that hydrate of the present invention remain with it is non- The biological effectiveness of the compound of hydrated form.
Unless otherwise mentioned, all suitable isotope variations of the compound of the present invention, stereoisomer, tautomerism Body, solvate, metabolite, salt and pharmaceutically acceptable prodrug are intended to be included within the scope of the present invention.
In structure disclosed by the invention, when the spatial chemistry of the chiral atom of any specific does not indicate, then the structure All stereoisomers all consider within the present invention, and be included in the invention as disclosed compound of present invention.When Spatial chemistry is expressed the real wedge-shaped line (solid wedge) of particular configuration or when dotted line indicates, then the alloisomerism of the structure Body is with regard to this clear and definition.
Compound shown in formula (I) can exist in a salt form.In one embodiment, the salt refers to and can pharmaceutically connect The salt received.Term " pharmaceutically acceptable " refer to substance or composition must with other ingredients comprising preparation and/or use it The mammal for the treatment of is compatible chemically and/or in toxicology.In another embodiment, the salt, which is not necessarily, pharmaceutically may be used The salt of receiving can be and be used to prepare and/or purify compound shown in formula (I) and/or for separating compound shown in this formula (I) Enantiomer intermediate.
Officinal salt of the invention can be synthesized with conventional chemical processes by parent compound, alkalinity or acidic moiety. In general, such salt can by make these compounds free acid form and stoichiometry suitable alkali (such as Na, Ca, Hydroxide, carbonate, bicarbonate of Mg or K etc.) reaction, or free alkali form and chemistry by making these compounds The suitable acid reaction of metered amount is to be prepared.Such reaction usually carries out in the mixture of water or organic solvent or both. Generally, in appropriate cases, it needs using non-aqueous medium such as ether, ethyl acetate, ethyl alcohol, isopropanol or acetonitrile.? Such as " Remington ' s Pharmaceutical Sciences ", the 20th edition, Mack Publishing Company, Easton,Pa.,(1985)；" pharmaceutical salts handbook: property, selection and application (Handbook of Pharmaceutical Salts:Properties, Selection, and Use) ", Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002) list that other is suitable for salt can be found in.
Any structural formula that the present invention provides, which is also intended to, indicates these compounds not by the form of isotope enrichment and same The form of position element enrichment.The structure that the general formula that there is the compound of isotope enrichment the present invention to provide is described, in addition to one or more A atom is replaced by the atom with selected atomic weight or mass number.The Exemplary isotopes that can be introduced into the compounds of this invention Isotope including hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur, fluorine and chlorine, such as2H、3H、11C、13C、14C、15N、17O、18O、18F、31P、32P、35S、36Cl and125I。
On the other hand, the present invention relates to the intermediates of compound shown in preparation formula (I).
On the other hand, the present invention provides a kind of pharmaceutical composition, and described pharmaceutical composition includes the compounds of this invention.One In embodiment, pharmaceutical composition of the present invention further includes pharmaceutically acceptable carrier, excipient, adjuvant, molten Matchmaker or their combination.In another embodiment, pharmaceutical composition can be liquid, solid, semisolid, gel or spray Type.
Such compound or its pharmaceutical composition of the invention can be used for the treatment of hepatitis C, have oral bioavailability The advantage that degree is high, metabolisming property is good, while having the function of protecting hepatic tissue, liver cell, improving liver function.
Specific embodiment
For the description present invention, it is listed below embodiment.But it is to be understood that the present invention is not limited to these Examples, only Method of the invention is practiced in offer.
Generally, the compound of the present invention described method can be prepared through the invention, unless there are further Explanation, wherein shown in the definition of substituent group such as formula (I).Following reaction scheme and embodiment is for being further illustrated this The content of invention.
The professional of fields will be appreciated that chemical reaction described in the invention can be used to suitably prepare perhaps Other compounds mostly of the invention, and other methods for the preparation of the compounds of the present invention are considered as in model of the invention Within enclosing.For example, the synthesis of the compound of those non-illustrations can be successfully by those skilled in the art according to the present invention It is completed by method of modifying, such as protection interference group appropriate, by utilizing other known reagent in addition to described in the invention , or reaction condition is made into some conventional modifications.In addition, reaction disclosed in this invention or known reaction condition are also generally acknowledged Ground is suitable for the preparation of other compounds of the invention.
The embodiments described below, unless other aspects show that all temperature are set to degree Celsius.Reagent purchase is in quotient Product supplier such as Aldrich Chemical Company, Arco Chemical Company and Alfa Chemical Company, all without by not being further purified when use, unless other aspects show.General reagent is from the western Gansu Province chemical industry in Shantou Factory, Guangdong Guanghua Chemical Reagent Factory, Guangzhou Chemical Reagent Factory, tianjin haoyuyu chemicals co., ltd., Tianjin good fortune morning chemistry Chemical reagent work, Wuhan Xin Huayuan development in science and technology Co., Ltd, Qingdao Tenglong Chemical Reagent Co., Ltd. and Haiyang Chemical Plant, Qingdao's purchase It can buy.
Anhydrous tetrahydro furan, dioxane, toluene, ether are dried to obtain by sodium metal reflux.Anhydrous methylene chloride It with chloroform is dried to obtain by calcium hydride reflux.Ethyl acetate, petroleum ether, n-hexane, n,N-dimethylacetamide and N, N- Dimethylformamide is used through anhydrous sodium sulfate is dry in advance.
Reaction is usually to cover a drying tube under positive pressure of nitrogen or argon or on anhydrous solvents (unless other aspects below Show), reaction flask all squeezed by syringe beyond the Great Wall by suitable rubber stopper, substrate.Glassware is all dried.
Chromatographic column is using silicagel column.Silica gel (300-400 mesh) is purchased from Haiyang Chemical Plant, Qingdao.
1H H NMR spectroscopy is recorded using Bruker 400MHz or 600MHz nuclear magnetic resonance spectrometer.1H H NMR spectroscopy is with CDC13、 DMSO-d6、CD3OD or acetone-d6For solvent (as unit of ppm), use TMS (0ppm) or chloroform (7.26ppm) as referring to mark It is quasi-.When there is multiplet, following abbreviation: s (singlet, unimodal), d (doublet, bimodal), t will be used (triplet, triplet), m (multiplet, multiplet), br (broadened, broad peak), dd (doublet of Doublets, double doublet), dt (doublet of triplets, double triplets).Coupling constant is indicated with hertz (Hz).
The determination condition of low resolution mass spectrometry (MS) data is: 6120 level four bars HPLC-M of Agilent (column model: Zorbax SB-C18,2.1x 30mm, 3.5 microns, 6min, flow velocity 0.6mL/min.Mobile phase: 5%-95% (contains 0.1% The CH of formic acid3CN) in (H containing 0.1% formic acid2O the ratio in), using electrospray ionisation (ESI), at 210nm/254nm, It is detected with UV.
Pure compound uses Agilent 1260pre-HPLC or Calesep pump 250pre-HPLC (pillar type Number: NOVASEP 50/80mm DAC), detected in 210nm/254nm with UV.
The use of logogram word below is through the present invention:
CH2Cl2, DCM methylene chloride；CDC13Deuterated chloroform；DMF N,N-dimethylformamide；DIPEA N, N- diisopropyl Base ethamine；DMSO dimethyl sulfoxide；MeOH methanol；MeCN,CH3CN acetonitrile；HCl hydrogen chloride；KI potassium iodide；t-BuOK Potassium tert-butoxide；NaHCO3Sodium bicarbonate；Na2S2O3Sodium thiosulfate；Na2SO4Sodium sulphate；10% palladium carbon of Pd/C；G grams；h Hour；ML, ml milliliters；PE petroleum ether (60-90 DEG C)；RT, rt, r.t. room temperature；Rt retention time；TFA trifluoroacetic acid.
Embodiment 1: the synthesis of compound II-A-1
Method A:
Step 1: 529mg (1.0mmol) III-1 is dissolved in 10mL anhydrous propanone, 680mg is added at room temperature (1.5mmol) IV-A-1 and potassium carbonate 414mg (3.0mmol), heating stirring reflux 6h, TLC detect fully reacting.Filtering, filter Liquid is diluted with methylene chloride (30mL), is successively washed with water (10mL) and saturated salt solution (10mL), and organic phase, anhydrous sulphur are separated It is filtered after sour sodium is dry, is concentrated under reduced pressure and removes solvent.Residue silica gel column chromatography purifies to obtain 710mg white solid V-A-1, yield 75%, ESI-MS:m/z [M+H]+=947.
Step 2: 200mg (0.21mmol) V-A-1 is dissolved in 10mL anhydrous methylene chloride, and 2mL (body is added at room temperature Product ratio: methylene chloride/tri isopropyl silane/trifluoroacetic acid=5/1/1) solution, reacts about 1h, TLC detects fully reacting, carefully Saturated sodium bicarbonate solution is added, separates organic phase saturated common salt water washing, anhydrous sodium sulfate depressurizes dense after drying, filtering Contracting.Residue silica gel column chromatography purifies to obtain 96mg white solid II-A-1, yield 65%.1H NMR(400MHz,DMSO-d6)δ 8.43 (t, J=5.7Hz, 1H), 7.68 (d, J=7.2Hz, 1H), 7.39 (t, J=7.7Hz, 2H), 7.30-7.14 (m, 3H), 6.14-5.96 (m, 2H), 5.90 (d, J=4.9Hz, 1H), 5.84 (d, J=9.6Hz, 1H), 5.81 (d, J=9.6Hz, 1H), 5.74 (d, J=8.2Hz, 1H), 4.87 (dt, J=12.4,6.2Hz, 1H), 4.47-4.34 (m, 1H), 4.29-4.20 (m, 1H), 4.08-4.01 (m, 1H), 3.94-3.76 (m, 4H), 3.57-3.46 (m, 1H), 2.82 (d, J=8.3Hz, 1H), 1.35 (d, J=6.9Hz, 3H), 1.27 (t, J=15.0Hz, 6H), 1.16 (d, J=6.2Hz, 6H)13C NMR(100MHz,DMSO- d6)δ173.7,173.1,173.0,169.2,161.1,151.2,151.1,150.6,130.1,125.1,120.6,120.5, 101.9,101.6,99.8,80.2,72.1,71.9,68.5,65.1,64.9,50.3,41.0,36.3,22.4,22.3,21.9, 21.8,20.3,20.2,17.2,16.9.31P NMR(162MHz,DMSO)δ3.83(s)ESI-MS:m/z[M+H]+=705.
Method B:
Step 1: 5.0g (9.5mmol) compound III-1 is dissolved in the anhydrous n,N-Dimethylformamide of 25mL (DMF) In, sequentially add 1.6g (23.5mmol) imidazoles under Ar protection, 0.12g (1.0mmol) 4-dimethylaminopyridine (DMAP) with And 1.6mL (9.5mmol) chlorotriethyl silane (TESCl).Reaction about 10 hours is stirred at room temperature, TLC detects fully reacting. Then 7mL 37%HCHO solution is added into reaction system, is heated to 80 degree and is stirred to react 1 hour.It is cooled to room temperature, is added 80mL ethyl acetate depressurizes dense after successively using water, saturated common salt water washing, separation organic phase to be dried, filtered with anhydrous sodium sulfate Contracting, obtains colorless oil VII-1, is directly used in without further purification in next step.
Step 2: the fresh resulting product VII -1 of previous step is dissolved in 40mL anhydrous methylene chloride (DCM), in argon gas It protects lower ice bath cooling, sequentially adds 62mg (0.5mmol) 4-dimethylaminopyridine (DMAP) and 2.36g (12.3mmol) 1- second Base-(3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC HCl) and 6.1g (11.4mmol) compound VIII-A- 1, it is slowly warmed to room temperature and is stirred to react about 12 hours, TLC detects fully reacting.100mL methylene chloride is added, successively with saturation Sodium bicarbonate aqueous solution, saturated common salt water washing, separation organic phase are concentrated under reduced pressure after being dried, filtered with anhydrous sodium sulfate.It is thick to produce Object recrystallizes to obtain 8.5g white solid product IX-A-1, yield 70% (two steps) with methylene chloride-normal heptane.1H NMR (400MHz,CDCl3) δ 7.57 (d, J=8.1Hz, 1H), 7.35 (dd, J=17.2,7.6Hz, 6H), 7.31-7.12 (m, 12H), 6.77 (d, J=8.5Hz, 2H), 6.17 (d, J=18.4Hz, 1H), 6.04-5.87 (m, 2H), 5.68 (d, J= 8.2Hz, 1H), 5.00 (dt, J=12.3,6.2Hz, 1H), 4.60 (dd, J=11.4,6.4Hz, 1H), 4.35 (q, J= 6.9Hz,1H),4.29–4.20(m,2H),4.15–4.08(m,1H),4.02–3.85(m,3H),3.78(s,3H),3.71(t,J =10.0Hz, 1H), 1.39-1.32 (m, 6H), 1.35 (s, 9H), 1.27 (d, J=11.9Hz, 3H), 1.23 (d, J=6.2Hz, 6H),0.92(s,9H),0.15(s,6H).ESI-MS:m/z[M+H]+=1191.
Step 3: 7.5g (6.3mmol) compound IX-A-1 is dissolved in 50% boiling (20mL) mixed solution, is added Enter 30mL glacial acetic acid (HOAc) and 6mL trifluoroacetic acid (TFA), is stirred to react at room temperature about 6 hours, until TLC can't detect residue Raw material.Vacuum rotary steam removes acetone, water, then sequentially adds 60mL methylene chloride, 9ml trifluoroacetic acid (TFA), and 6.5mL tri- is different Propyl silane (i-Pr3SiH), the reaction was continued at room temperature about 15 minutes, and TLC detects fully reacting, is carefully added into unsaturated carbonate hydrogen Sodium water solution extracts reaction of going out.100mL methylene chloride is added, successively uses water, saturated common salt water washing, separates the anhydrous sulphur of organic phase Crude product is concentrated under reduced pressure to obtain in sour sodium after drying, filtering.3.9g white solid II-A-1 is recrystallized to obtain with ethyl acetate-light petrol, Yield 88%.1H NMR (400MHz, DMSO) δ 8.43 (t, J=5.7Hz, 1H), 7.68 (d, J=7.2Hz, 1H), 7.39 (t, J =7.7Hz, 2H), 7.30-7.14 (m, 3H), 6.14-5.96 (m, 2H), 5.90 (d, J=4.9Hz, 1H), 5.84 (d, J= 9.6Hz, 1H), 5.81 (d, J=9.6Hz, 1H), 5.74 (d, J=8.2Hz, 1H), 4.87 (dt, J=12.4,6.2Hz, 1H), 4.47–4.34(m,1H),4.29–4.20(m,1H),4.08–4.01(m,1H),3.94–3.76(m,4H),3.57–3.46(m, 1H), 2.82 (d, J=8.3Hz, 1H), 1.35 (d, J=6.9Hz, 3H), 1.27 (t, J=15.0Hz, 6H), 1.16 (d, J= 6.2Hz,6H).ESI-MS:m/z[M+H]+=705.
Recrystallization mother liquor is concentrated, residue purifies to obtain a small amount of white solid II-A-9 using silica gel column chromatography,1H NMR(400MHz,DMSO)1H NMR(400MHz,CDCl3) δ 7.51 (d, J=7.9Hz, 1H), 7.35 (t, J=7.8Hz, 2H), 7.20 (dd, J=14.5,7.5Hz, 3H), 6.19 (brs, 1H), 6.07 (d, J=9.4Hz, 1H), 5.99 (d, J=9.4Hz, 1H), 5.72 (d, J=8.2Hz, 1H), 5.00 (dt, J=12.5,6.2Hz, 1H), 4.59-4.39 (m, 2H), 4.37-4.22 (m, 2H), 4.14 (d, J=8.6Hz, 1H), 3.99-3.85 (m, 3H), 3.65-3.58 (m, 1H), 1.48 (d, J=7.0Hz, 1H), 1.42-1.36 (m, 3H), 1.38-1.26 (m, 6H), 1.24 (d, J=6.2Hz, 6H) .ESI-MS:m/z [M+H]+= 705。ESI-MS:m/z[M+H]+=705.
Embodiment 2: the synthesis of compound II-A-1a
According to the fresh obtained compound VII-1 of method step B one in embodiment 1, (3mmol compound III-1 is that starting is former Material), it is dissolved in 12mL anhydrous methylene chloride (DCM), ice bath is cooling under protection of argon gas, sequentially adds 18mg (0.15mmol) 4- Dimethylamino naphthyridine (DMAP) and 0.75g (3.9mmol) 1- ethyl-(3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDC HCl) and 1.93g (3.6mmol) compound VIII-A-1a (are prepared, R configuration chiral purity by chiral Tiopronin Degree 95%, analysis method: AD-H column temperature: 25C；Detection wavelength: 210nm；Flow velocity: 1.0ml/min；Mobile phase: n-Hex:EtOH: TFA=90:10:0.1 isocratic elution), it is then slowly warmed to room temperature and is stirred to react about 12 hours, TLC detects fully reacting.Add Enter 30mL methylene chloride, successively use saturated sodium bicarbonate aqueous solution, saturated common salt water washing, separates organic phase anhydrous sodium sulfate It is concentrated under reduced pressure after drying, filtering.Crude product recrystallizes to obtain 8.7g white solid product IX-A-1a with methylene chloride-normal heptane, receives Rate 73%.
2.1g (1.76mmol) compound IX-A-1a is dissolved in 50% boiling (5mL) mixed solution, 8mL ice is added Acetic acid (HOAc) and 2mL trifluoroacetic acid (TFA) are stirred to react about 6 hours, at room temperature until TLC can't detect surplus stock.Decompression Revolving removes acetone, water, then sequentially adds 15mL methylene chloride, 3ml trifluoroacetic acid (TFA), 2.2mL tri isopropyl silane (i-Pr3SiH), the reaction was continued at room temperature about 15 minutes, and TLC detects fully reacting, and it is molten to be carefully added into saturated sodium bicarbonate water Liquid extracts reaction of going out.30mL methylene chloride is added, successively uses water, saturated common salt water washing, separation organic phase is dry with anhydrous sodium sulfate It is dry, crude product is concentrated under reduced pressure to obtain after filtering.1.7g white solid II-A-1a, yield are recrystallized to obtain with ethyl acetate-light petrol 86%.ESI-MS:m/z[M+H]+=705.
Embodiment 3: the synthesis of compound II-A-1b
Step 1: step 2, (is prepared, S with VIII-A-1b by chiral Tiopronin in foundation 1 method B of embodiment Configuration chiral purity 97%, analysis method: AD-H column temperature: 25C；Detection wavelength: 210nm；Flow velocity: 1.0ml/min；Mobile phase: N-Hex:EtOH:TFA=90:10:0.1 isocratic elution) VIII-A-1a is replaced, it can be made with 5.3g (10mmol) compound III-1 It is standby to obtain 2.57g intermediate compound I X-A-1b, yield 72% (two steps).
According to step 3 in 1 method B of embodiment, compound IX-A-1b (2.9mmol) continues to slough protecting group, can prepare Obtain 1.04g white solid product II-A-1b, yield 84%.ESI-MS:m/z[M+H]+=705.
Embodiment 4: the synthesis of compound II-A-2
Method A:
Step 1: 529mg (1.0mmol) III-1 is dissolved in 10mL anhydrous propanone, 680mg is added at room temperature (1.5mmol) IV-A-2 and potassium carbonate 414mg (3.0mmol), heating stirring reflux 6h, TLC detect fully reacting.Filtering, filter Liquid is diluted with methylene chloride (30mL), is successively washed with water (10mL) and saturated salt solution (10mL), and organic phase, anhydrous sulphur are separated It is filtered after sour sodium is dry, is concentrated under reduced pressure and removes solvent.Residue silica gel column chromatography purifies to obtain 306mg white solid V-A-2.ESI- MS:m/z[M+H]+=961.
Step 2: 200mg (0.21mmol) V-A-2 is dissolved in 10mL anhydrous methylene chloride, and 2mL (body is added at room temperature Product ratio: methylene chloride/tri isopropyl silane/trifluoroacetic acid=5/1/1) solution, reacts about 1h, TLC detects fully reacting, carefully Saturated sodium bicarbonate solution is added, separates organic phase saturated common salt water washing, anhydrous sodium sulfate depressurizes dense after drying, filtering Contracting.Residue silica gel column chromatography purifies to obtain 100mg white solid II-A-2, yield 67%.1H NMR(400MHz,CDCl3)δ 9.64 (s, 1H), 7.50 (dd, J=8.1,2.3Hz, 1H), 7.36 (t, J=7.8Hz, 2H), 7.30-7.22 (m, 3H), 7.19 (d, J=7.6Hz, 1H), 6.65 (q, J=6.5Hz, 1H), 6.19 (d, J=17.9Hz, 1H), 5.10-4.94 (m, 1H), 4.52 (dd, J=11.4,5.7Hz, 1H), 4.39-4.21 (m, 3H), 4.22-4.05 (m, 2H), 3.98 (tt, J=11.5,5.8Hz, 1H), 3.57-3.45 (m, 1H), 2.17 (dd, J=8.5,2.2Hz, 1H), 1.86 (d, J=6.5Hz, 3H), 1.56 (dd, J= 7.1,1.0Hz, 3H), 1.37 (dd, J=14.8,7.5Hz, 6H), 1.27 (s, 1H), 1.25 (d, J=6.3Hz, 6H) .ESI- MS:m/z[M+H]+=719.
Method B:
According to 1 method step B one of embodiment, step 2, formaldehyde is replaced with acetaldehyde, with 5.3g (10mmol) compound III- 1 can be prepared into 4.6g intermediate compound I X-A-2, yield 38% (two steps).ESI-MS:m/z[M+H]+=1205.
According to 1 method step B three of embodiment, compound IX-A-2 (2.9mmol) continues to slough protecting group, can be prepared into 1.7g white solid product II-A-2, yield 82%.ESI-MS:m/z [M+H]+=719.
Embodiment 5: the synthesis of compound II-A-3
According to 1 method step B one of embodiment, step 2, formaldehyde is replaced with propionic aldehyde, with 2.65g (5mmol) compound III- 1 can be prepared into 2g intermediate compound I X-A-3, yield 33% (two steps).ESI-MS:m/z[M+H]+=1219.
According to 1 method step B three of embodiment, compound IX-A-3 (0.8mmol) continues to slough protecting group, can be prepared into 445mg white solid product II-A-3, yield 76%.ESI-MS:m/z [M+H]+=733.
Embodiment 6: the synthesis of compound II-A-4
Step 1: 529mg (1.0mmol) III-1 is dissolved in 10mL anhydrous propanone, 690mgIV-A-3 is added at room temperature And potassium carbonate 414mg (3.0mmol), heating stirring reflux 6h, TLC detect fully reacting.Filtering, filtrate methylene chloride (30mL) dilution, is successively washed with water (10mL) and saturated salt solution (10mL), separates organic phase, mistake after anhydrous sodium sulfate is dry Filter is concentrated under reduced pressure and removes solvent.Residue silica gel column chromatography purifies to obtain 300mg V-A-4, is directly used in and reacts in next step.
Step 2: 200mg (0.21mmol) V-A-4 is dissolved in 10mL anhydrous methylene chloride, and 2mL (body is added at room temperature Product ratio: methylene chloride/tri isopropyl silane/trifluoroacetic acid=5/1/1) solution, reacts about 1h, TLC detects fully reacting, carefully Saturated sodium bicarbonate solution is added, separates organic phase saturated common salt water washing, anhydrous sodium sulfate depressurizes dense after drying, filtering Contracting.Residue silica gel column chromatography purifies to obtain 100mg white solid II-A-4.1H NMR(400MHz,CDCl3)δ9.62(s, 1H), 7.46 (dd, J=8.1,2.2Hz, 1H), 7.34 (t, J=7.8Hz, 2H), 7.29-7.22 (m, 3H), 7.19 (d, J= 7.4Hz, 1H), 6.16 (d, J=18.1Hz, 1H), 5.64-5.53 (m, 1H), 5.37-5.17 (m, 1H), 5.08-4.93 (m, 1H), 4.53 (dd, J=11.4,5.5Hz, 1H), 4.38-4.20 (m, 3H), 4.40-4.08 (m, 6H), 3.97 (tt, J= 11.7,5.9Hz, 1H), 3.57-3.43 (m, 1H), 2.17 (dd, J=8.4,2.2Hz, 1H), 1.55 (dd, J=7.1,1.0Hz, 3H), 1.38 (dd, J=14.8,7.7Hz, 6H), 1.27 (s, 1H), 1.24 (d, J=6.3Hz, 6H) .ESI-MS:749 (M+1).
Embodiment 7: the synthesis of compound II-A-5
Step 1: 529mg (1.0mmol) III-1 is dissolved in 10mL anhydrous propanone, and 800mg IV-A-4 is added at room temperature And potassium carbonate 414mg (3.0mmol), heating stirring reflux 12h, TLC detect fully reacting.Filtering, filtrate methylene chloride (30mL) dilution, is successively washed with water (10mL) and saturated salt solution (10mL), separates organic phase, mistake after anhydrous sodium sulfate is dry Filter is concentrated under reduced pressure and removes solvent.Residue silica gel column chromatography purifies to obtain 400mg V-A-5, is directly used in and reacts in next step.
Step 2: 200mg (0.21mmol) V-A-5 is dissolved in 10mL anhydrous methylene chloride, and 2mL (body is added at room temperature Product ratio: methylene chloride/tri isopropyl silane/trifluoroacetic acid=5/1/1) solution, reacts about 1h, TLC detects fully reacting, carefully Saturated sodium bicarbonate solution is added, separates organic phase saturated common salt water washing, anhydrous sodium sulfate depressurizes dense after drying, filtering Contracting.Residue silica gel column chromatography purifies to obtain 100mg white solid II-A-5.1H NMR(400MHz,CDCl3)δ9.66(s, 1H), 7.44 (dd, J=7.9,2.1Hz, 1H), 7.32 (t, J=7.8Hz, 2H), 7.28-7.20 (m, 3H), 7.16 (d, J= 7.5Hz, 1H), 6.13 (d, J=17.6Hz, 1H), 5.15-5.03 (m, 1H), 4.60 (t, J=7.5Hz, 2H), 4.55 (dd, J =11.5,5.7Hz, 1H), 4.48-4.25 (m, 3H), 4.23-4.09 (m, 2H), 3.95 (tt, J=11.8,5.8Hz, 1H), 3.59-3.42 (m, 1H), 3.25 (t, J=7.5Hz, 2H), 2.16 (dd, J=8.4,2.2Hz, 1H), 1.54 (dd, J=7.1, 1.0Hz, 3H), 1.39 (dd, J=14.7,7.6Hz, 6H), 1.28 (s, 1H), 1.25 (d, J=6.5Hz, 6H) .ESI-MS:m/z [M+H]+=719.
Embodiment 8: the synthesis of compound II-A-7
According to 1 method step B one of embodiment, step 2, III-1 is only replaced with III-2, with 1.1g (2mmol) change Colorless oil IX-A-7, yield 71% (two steps) can be prepared by closing object III-2.ESI-MS:m/z[M+H]+=1207.
According to 1 method step B three of embodiment, 0.79g compound IX-A-7 (0.65mmol) continues to slough protecting group, can make It is standby to obtain white solid product II-A-7, yield 83%.ESI-MS:m/z[M+H]+=721.
Embodiment 9: the synthesis of compound II-A-8
According to 1 method step B one of embodiment, step 2 replaces VIII-A-1 with VIII-A-2, thus with 1.6g (3mmol) compound III-1 can be prepared into 1.29g intermediate compound I X-A-8, yield 65% (two steps).ESI-MS:m/z[M+H]+= 1219.
According to 1 method step B three of embodiment, compound IX-A-8 (0.8mmol) continues to slough protecting group, can be prepared into 0.5g white solid product II-A-8, yield 83%.ESI-MS:m/z [M+H]+=733.
Embodiment 10: the synthesis of compound II-B-1
According to 1 method B of embodiment, in generation, (is prepared) by (DL)-N-acetylcystein with VIII-B-1 in step 2 For VIII-1,3.38g intermediate compound I X-B-1, yield 71% (two steps) can be prepared into 2.1g (4mmol) compound III-1. ESI-MS:m/z[M+H]+=1191.
According to 1 method step B three of embodiment, compound IX-B-1 (2mmol) continues to slough protecting group, can be prepared into 1.17g white solid product II-B-1, yield 83%.ESI-MS:m/z [M+H]+=705.
Embodiment 11: the synthesis of compound II-B-1a
According to 1 method B of embodiment, (prepared by L- acetylcysteine, chiral purity in step 2 with VIII-B-1a Degree > 99%) VIII-1 is replaced, intermediate compound I X-B-1a, yield 67% (two can be prepared into 2.65g (5mmol) compound III-1 Step).ESI-MS:m/z[M+H]+=1191.
According to 1 method step B three of embodiment, compound IX-B-1a (2.1mmol) continues to slough protecting group, can be prepared into 1.2g white solid product II-B-1a, yield 85%.ESI-MS:m/z [M+H]+=705.
Embodiment 12: the synthesis of compound II-B-1b
According to 1 method B of embodiment, (prepared by D-Cys, chiral purity in step 2 with VIII-B-1b 95%) VIII-1 is replaced, 4.2g intermediate compound I X-B-1b, yield 70% can be prepared into 2.65g (5mmol) compound III-1 (two steps).ESI-MS:m/z[M+H]+=1191.
According to 1 method step B three of embodiment, compound IX-B-1b (2.1mmol) continues to slough protecting group, can be prepared into 1.2g white solid product II-B-1b, yield 81%.ESI-MS:m/z[M+H]+=705.
Embodiment 13: the synthesis of compound II-B-2
According to 1 method step B one of embodiment, formaldehyde is replaced with acetaldehyde, according to 1 method step B two of embodiment, with VIII- B-1a (being prepared by L- acetylcysteine, chiral purity > 99%) replaces VIII-1, thus with 1.6g (3mmol) chemical combination Object III-1 can be prepared into 1.3g intermediate compound I X-B-2, yield 36% (two steps).ESI-MS:m/z[M+H]+=1205.
According to 1 method step B three of embodiment, compound IX-B-2 (1mmol) continues to slough protecting group, can be prepared into 0.57g white solid product II-B-2, yield 79%.ESI-MS:m/z[M+H]+=719.
Embodiment 14: the synthesis of compound II-B-3
According to 1 method step B one of embodiment, step 2 (is prepared, hand with VIII-B-2 by L- acetylcysteine Property purity > 99%) replace VIII-1, so that intermediate compound I X-B-3 can be prepared into 1.6g (3mmol) compound III-1, yield 76% (two steps).ESI-MS:m/z[M+H]+=1205.
According to 1 method step B three of embodiment, compound IX-B-3 (1mmol) continues to slough protecting group, can be prepared into 0.57g white solid product II-B-3, yield 82%.ESI-MS:m/z[M+H]+=719.
Embodiment 15: the synthesis of compound II-B-4
According to step 1 in 1 method B of embodiment, step 2 replaces III-1 with III-2, and with VIII-B-1a (by L- second Acyl cysteine prepares, chiral purity > 99%) VIII-1 is replaced, it can be prepared into 1.6g (3mmol) compound III-2 2.64g intermediate compound I X-B-4, yield 72% (two steps).ESI-MS:m/z[M+H]+=1207.
According to 6 step 3 of embodiment, compound IX-B-4 (1.5mmol) continues to slough protecting group, and it is white can be prepared into 0.93g Color solid product II-B-4, yield 84%.ESI-MS:m/z [M+H]+=721.
Embodiment 16: anti-hepatitis C virus activity (HCV, EC50) and cytotoxicity (CC50)
Due to lacking ideal HCV Infection in Vitro cell and animal model, under normal circumstances, HCV virus activity rating meeting Extracellular molecule reconstructed model is established using the enzyme to play a crucial role in HCV rna replicon.
Experimental method:
Containing 10% fetal calf serum, 1X nonessential amino acid, Pen-Strep-Glu, G418 DMEM culture medium in, training Support the Huh7 cell of replicon containing HCV- (1b genotype).Antiviral screening test without G418 different culture medium in into Row.The compound of test is added in cell inoculation immediately after 96 orifice plates, inoculating cell, and 37 DEG C of incubations in the incubator.Then Culture medium is removed, cell is used for full nucleic acid extraction (including replicon rna and host RNA).It can amplify in Q-RT-PCR scheme Replicon rna, and it is accordingly quantitative.The level difference of observed replicon HCV RNA and untreated control group are as test The mode of compound antiviral efficacy, the result is shown in tables 1.
Anti- hepatitis C virus activity (HCV, the EC of 1 compound of table50) and cytotoxicity (CC50) result
Inhibitory activity (the EC of HCV replicon 1b in compound on intracellular50) result it is living as its anti-hepatitis C virus is evaluated Property, and cytotoxic activity (CC simultaneously50) synchronism detection for exclude the false positive results as caused by cytotoxicity.Treatment refers to Number is higher, and representation compound more has application prospect.The result shows that compound provided by the invention is shown and Sofosbuvir phase Like or more preferably activity and therapeutic index, by taking II-A-1, II-A-1a, II-A-1b, II-A-2 as an example, to intracellular HCV Inhibitory activity (the EC of replicon50) be better than than positive control Sofosbuvir, show that the compound can be in liver cell Sofosbuvir it is the same can in the cell fast degradation become active metabolite form play HCV-Ab IgG effect, in addition, Another one's share of expenses for a joint undertaking Tiopronin degraded in molecule has antiviral activity certain gain effect, and this synergistic effect Make compound provided by the invention, application prospect is had more in terms of HCV-Ab IgG disease.
Embodiment 17: mouse CCl4 liver damages the protective effect research of model
Experimental method: it before taking the male mice of 24~28g of weight to test, is deprived of food but not water 6 hours, is randomly divided by weight 3 groups, every group of 5 mouse set up blank control group, CCL separately4Group, compound group (Tiopronin 50mg/kg, test compound 200mg/kg).Every 12h gastric infusion 1 time, each stomach-filling capacity is 10mL/kg, is administered 4 times altogether.Blank control group and model Control group gavages the physiological saline for waiting capacity simultaneously.The 1h after third time is administered is other two groups small in addition to blank control group mouse Mouse presses the CCl of 2mg/kg b.w450%CCl is injected intraperitoneally4Corn oil.After 12 hours, each group is administered once again.It is small in 24 Shi Hou, each mouse take blood, measure Serum ALT and AST (table 2), take liver to cut fritter, after formaldehyde is fixed, paraffin section uses hematoxylin And eosin stains, microscopically observation liver cell situation.
The protective effect result of study of the CCl4 liver damage model of 2 the compounds of this invention of table
The result shows that CCl4The ALT and AST of model group mouse are increased extremely, and Tiopronin can significantly reverse this liter of enzyme Effect, compared under relatively, the Sofosbuvir then not no activity of this respect.And compound provided by the invention has apparent drop enzyme Effect, activity are quite or more excellent with Tiopronin.It traces it to its cause, it may be possible to because the carboxyl of Tiopronin is made into prodrug Afterwards, its stability and the ability by oral absorption can be improved, so that its activity be made to be enhanced.With II-A-1, II-A-a, For II-A-2, II-A-5, causes ALT and AST raising to act on CCl4 induced liver injury, there is apparent inhibitory activity, and Better than positive control Tiopronin.In addition, liver organization pathology section examination result is CCl4Model group, central veins of liver become silted up Blood, around there is centrilobular or a peripheral necrosis, and liver cell has swelling, the deformation of balloon sample, II-A-1, II-A-a, II-A-2, A small amount of dotted and piecemeal necrosis is presented in the administration groups such as II-A-5, most of that the deformation of balloon sample is presented, and shows by CCl4It is caused Toxicity has greatly reduced, and illustrating compound of the present invention really has protection CCl4Caused liver harm is used.
Due to lacking ideal HCV infection animal model, directly evaluates compound provided by the invention and cause liver to damage HCV The protective effect of wound is infeasible, but theoretically and from the perspective of clinical application, the liver protection shield of compound provided by the invention The effect of liver equally has protective effect for inflammatory reaction caused by HCV.
Embodiment 18: pharmacokinetic property research
1 experimental material
1.1 compound
The compound of the present invention II-A-1, II-A-1a, II-A-1b, II-A-2, the II-B- prepared using above embodiments 1, III-2A-1, III-2A-1a (30mg/kg, the amount and Suo Fei group of nucleoside moiety are consistent), positive control drug Suo Feibuwei Each compound is added to 5%DMSO+60%PEG400+35% physiological saline by (25mg/kg), and it is mixed that 10mg/ml is prepared into after vortex Suspension is used for gastric infusion.
GS331007 is bought from splendid remote scientific and technological (Shanghai) Co., Ltd..
1.2 experimental animals and sampling
Using SD rat as experimental animal (weight 180-220g), 6 SD rats of each test compound are randomly divided into 2 groups (stomach-filling group and intravenous group), every group 3.It is 0.17,0.33,0.5,1,1.5,2,4 that the tail vein of gastric infusion, which takes blood time point, 6,8,12,24 hours；It is 0.05,0.1,0.17,0.5,1,2,4,6,8,12,24 hours that intravenously administrable, which takes blood time point,.It takes complete Blood 0.3ml takes blood plasma 0.1ml to be analyzed using LC-MS after centrifugation.
1.3 instrument
1200 liquid chromatographic system of Agilent, API4000QTRAP triple quadrupole bar tandem mass spectrometer
1.4 sample treatment
30 μ l liver homogenate samples or standard curve sample are taken to be added into the acetonitrile of 100 μ l containing the internal standards (100ng/ml), 10min (6000 turns/min) are centrifuged after vortex 2min, supernatant is transferred in sample injection bottle；
Take 5 μ L supernatants, sample introduction, using the concentration of GS331007 in LC-MS/MS test sample.It is even according to resulting liver Drug concentration-versus-time data are starched, are calculated separately respectively using the non-compartment model of pharmacokinetics software for calculation WinNonlin6.2.1 The pharmacokinetic parameter for testing the metabolin GS331007 of compound, the results are shown in Table 3.
Main pharmacokinetic parameters summarize after the administration of 3 SD Oral Administration in Rats of table
According to the literature, metabolic pathway in vivo, wherein GS331007 is final metabolite, is research Sofosbuvir in vivo bioavilability and metabolism behavior marker.Compound II-A-1, II-A-1a, II-A-1b, II- A-2, II-B-1 GS331007 blood concentration and metabolism behavior in rat body show the metabolite of test-compound GS331007 blood concentration and area under the curve have obviously compared with the blood concentration of GS331007 in Suo Feibuwei group rat body Improve, in other words, compound of the present invention improves obviously in terms of oral administration biaavailability.With compound II-A-1, II- For A-2, II-B-1, oral administration biaavailability is the 3.38 of Suo Feibuwei, 2.75,2.25 times respectively.In addition, II-A-1 and Its half-life period of II-A-2 has respectively reached 6.14 and 6.23 hours, respectively the 2.95 and 2.51 of Suo Feibuwei times, when medicine Curve more mitigates.Therefore, compound of the present invention is with the obvious advantage on pharmacokinetics, can be further used for taking orally to The treatment of medicine progress hepatitis disease.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and Made any modifications, equivalent replacements, and improvements etc., is all included in the scope of protection of the present invention within principle.

Claims (14)

1. compound has structure or its tautomer, optical isomer, pharmaceutically acceptable salt shown in Formula II-A:
Wherein, X is hydroxyl or F, Cl, Br；
Linker isOr it is not present；
R1For H or C1-C5Alkyl；N is 1,2,3,4 or 5；R3、R4It is each independently H or C1-C5Alkyl.
2. compound according to claim 1, with structure shown in Formula II-Aa or Formula II-Ab or its mutually variation Structure body, optical isomer, pharmaceutically acceptable salt:
Wherein, X is hydroxyl or F, Cl, Br；
Linker isOr it is not present；
R1For H or C1-C5Alkyl；N is 1,2,3,4 or 5；R3、R4It is each independently H or C1-C5Alkyl.
3. compound according to claim 1 is structure shown in Formula II-A-A:
Wherein,
X be hydroxyl or F, Cl, Br,
Linker is
R1For H or C1-C5Alkyl, n are the integer of 1-19.
4. compound according to claim 3 is Formula II-A-Aa, Formula II-A-Ab, Formula II-A-Ac, Formula II-A-Ad difference Shown in structure:
Wherein, X is hydroxyl or F, Cl, Br；R1For H or C1-C5Alkyl, n 1,2,3,4 or 5.
5. compound according to claim 4 is Formula II-A-Aa1, Formula II-A-Ab1, Formula II-A-Ac1, Formula II-A- The structure that Ad1 is respectively shown in:
Wherein, R1For H or C1-C5Alkyl, n 1,2,3,4 or 5.
It is mutual with structure as shown below or structural compounds as shown below 6. compound according to claim 1 Tautomeric, pharmaceutically acceptable salt:
7. a kind of preparation method of Formula II-A compound represented, step are as follows:
1) formula III compound reacts to obtain Formula V-A compound with formula IV-A compound,
2) Formula V-A compound obtains Formula II-A compound by deprotection reaction；
Wherein, X is hydroxyl or F, Cl, Br；
Linker isOr it is not present；
N is 1,2,3,4 or 5；R1For H or C1-C5Alkyl；R3、R4It is each independently hydrogen atom or C1-C5Alkyl；LG is leaving group Group；Z is thiol protecting group.
8. a kind of preparation method of Formula II-A-Aa compound represented can be passed through by compound of formula III and compound of Formula IV It crosses alkylation reaction and obtains general formula V-A compound, then obtained by deprotection reaction.
Wherein, X F, Cl or Br, R1For H or C1-C5Alkyl；Y is leaving group, and Z is thiol protecting group.
9. a kind of preparation method of Formula II-A-Ab compound represented, can be by compound of formula III and general formula IV-B compound General formula V-B compound is obtained by alkylation reaction, is then obtained by deprotection reaction:
Wherein, X is hydroxyl or F, Cl, Br；R1For H or C1-C5Alkyl；N is the integer of 1-19；Y is leaving group；
Z is thiol protecting group.
10. a kind of preparation method of Formula II-A-B compound, step are as follows:
1) formula III compound is reacted through overprotection, and Formula VII compound is then obtained with aldehyde reaction；
2) Formula VII compound and VIII-A compound react to obtain Formula IX-A-B compound；
3) Formula IX-A-B compound sloughs protecting group and obtains Formula II-A-B compound；
Wherein, X is hydroxyl or F, Cl, Br；R1For H or C1-C5Alkyl；R3、R4It is each independently hydrogen atom or C1-C5Alkyl；P1 For hydroxy-protective group,；P2For amide NH blocking group；P3For thiol protecting group.
11. a kind of compound, with structure shown in Formula IX-A-B or salt:
Wherein X is hydroxyl or F, Cl, Br；R1For H or C1-C5Alkyl；P1For hydroxy-protective group；P2For NH blocking group；P3For Thiol protecting group.
12. a kind of include one of compound shown in claim any one of 1-6 or a variety of pharmaceutical compositions.
13. pharmaceutical composition according to claim 12, wherein Formula II-A compound represented or its tautomer, Dosage of the one or more pharmaceutical compositions of pharmaceutically acceptable salt in described pharmaceutical composition is 1~1000mg/ It.
14. composition described in compound or claim 12 described in any one of claim 1~6, in preparation as treatment Purposes in hepatitis C drug.
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104936970A (en) * 2012-10-08 2015-09-23 埃迪尼克斯医药公司 2'-chloro nucleoside analogs for HCV infection

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Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104936970A (en) * 2012-10-08 2015-09-23 埃迪尼克斯医药公司 2'-chloro nucleoside analogs for HCV infection

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
《A Versatile One-Pot Procedure to Phosphate Monoesters and Pyrophosphates Using Di(p-methoxybenzyl)-N,N-diisopropylphosphoramidite》;Gerbrand J;《organic letters》;20080919;第10卷(第20期);全文
《射用硫普罗宁治疗慢性肝炎的临床观察》;李竹轩;《工企医刊》;20050331;第18卷(第3期);全文

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