CN108250121A

CN108250121A - Sulfonamide-arylamides and its medicinal usage for treating hepatitis B

Info

Publication number: CN108250121A
Application number: CN201611237327.6A
Authority: CN
Inventors: 王喆; 王晓光; 卢涔宾; 范国钦
Original assignee: Shanghai Longwood Pharmaceutical Co Ltd
Current assignee: Shanghai Longwood Pharmaceutical Co Ltd
Priority date: 2016-12-28
Filing date: 2016-12-28
Publication date: 2018-07-06
Also published as: CN108250122B; WO2018121689A1; CN108250122A

Abstract

The present invention relates to a kind of sulfonamide arylamides and its medicinal usages for the treatment of hepatitis B.Specifically, the invention discloses a kind of compounds with structure shown in chemical formula (A) as hbv replication inhibitor, or its stereoisomer, cis-trans-isomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, the definition of each group refer to specification.Purposes the invention further relates to the pharmaceutical composition comprising above compound and its in hepatitis B is treated.

Description

Sulfonamide-arylamides and its medicinal usage for treating hepatitis B

Technical field

The invention belongs to chemical medicine, in particular it relates to a kind of sulfonamide-arylamides and It treats the medicinal usage of hepatitis B.

Background technology

Hepatitis type B virus (HBV) is a kind of tunicary, dsdna segment (dsDNA), Hepadna Virus DNA families The virus of (hepatovirus section (Hepadnaviridae)).Its genome includes 4 overlapping open reading frames：Pronucleus/karyogene, polymerization Enzyme gene, UM and S genes (they encode three envelope proteins) and X gene.When before infection, the dsdna segment base Because of group in host cell nuclear (open-circle DNA；RcDNA) it is changed into covalently closed circular DNA (cccDNA) and virus mRNA It is transcribed.Once by encapsidate, which uses as template In reverse transcription, this reverse transcription regenerates part dsDNA genomes (rcDNA) in nucleocapsid.

HBV causes epidemic disease, and it is endemic in China in some areas in Asia and Africa.HBV is About 2,000,000,000 people are infected in the whole world, wherein about 3.5 hundred million people are developed into chronic infectious disease.The virus causes hepatitis B To increase risk associated for disease and chronic infectious disease and the development of hepatic sclerosis and liver cancer high.

The Spreading source of hepatitis type B virus has in serum efficient in being exposed to communicable blood or body fluid Viral DNA is detected in the saliva of the chronic carriers of valency DNA, tear and urine.

Although presently, there are vaccine a kind of effective and with good tolerability, the selection directly treated is current It is also limited to interferon and following antiviral agent；Tenofovir, tenofovir Chinese mugwort draw phenol amine (TAF), Lamivudine, A Defu Wei, Entecavir and Sebivo.

Press down in addition, heteroaryl dihydro-pyrimidin (HAPs) is authenticated in tissue cultures and animal model as a kind of HBV Preparation (weber (Weber) et al.,《Antiviral study》(Antiviral Res.)54:69-78).

WO 2013/006394 (being disclosed on January 10th, 2013) and WO 2013/096744 (are disclosed in June 27 in 2013 Day) also disclose the sulfamoyl-aryl amide for being related to Anti-HBV effect.

However, can be encountered in these direct HBV antiviral agents toxicity, mutagenicity, lack selectivity, weak curative effect, The problems such as poor bioavailability and difficult synthesis.

Therefore, in order to overcome with upper curve, need exploitation that there is the HBV inhibitor that potency is high, toxicity is more low.

Invention content

It can be used as the compound of the structure novel of HBV inhibitor the object of the present invention is to provide one kind.

First aspect present invention provide it is a kind of as compound shown in formula A or its stereoisomer, cis-trans-isomer or Tautomer or its pharmaceutically acceptable salt, hydrate or solvate,

Wherein,

R1, R2 are each independently hydrogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C3-C10 cycloalkanes Base, substituted or unsubstituted heteroatomic 3-10 membered heterocycloalkyls, the substitution or unsubstituted for being selected from the group N, S and O with 1-3 C6-C10 aryl, the substituted or unsubstituted heteroatomic 5-10 unit's heteroaryls that N, S and O are selected from the group with 1-3；Or R1, R2 with and their nitrogen-atoms for being connected collectively form and substituted or unsubstituted there are 1 N and 0-3 to be selected from the group N, S and O Heteroatomic 3-10 membered heterocycloalkyls；

R3 for hydrogen, halogen ,-CN, hydroxyl, amino, carboxyl ,-(C=O)-substituted or unsubstituted C1-C8 alkyl, substitution or Unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkylamino radicals, substituted or unsubstituted C1-C8 alkoxyl, substitution or Unsubstituted C3-C10 cycloalkyl, the substituted or unsubstituted heteroatomic 3-10 members that N, S and O are selected from the group with 1-3 are miscellaneous Cycloalkyl, substituted or unsubstituted C6-C10 aryl and the substituted or unsubstituted miscellaneous original for being selected from the group N, S and O with 1-3 The 5-10 unit's heteroaryls of son；

R4, R5 and R6 are the substituent group of any position on phenyl ring, are each independently hydrogen, halogen ,-CN, hydroxyl, ammonia Base, carboxyl ,-(C=O)-substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkyl, substitution or unsubstituted C2-C6 alkenyls, substituted or unsubstituted C2-C6 alkynyls, substituted or unsubstituted C1-C8 alkylamino radicals, substituted or unsubstituted C1-C8 alkoxyl, substituted or unsubstituted there is 1-3 to be selected from the group N, S and O at substituted or unsubstituted C3-C10 cycloalkyl Heteroatomic 3-10 membered heterocycloalkyls, substituted or unsubstituted C6-C10 aryl and it is substituted or unsubstituted have 1-3 select From the heteroatomic 5-10 unit's heteroaryls of the following group N, S and O；

In R1, R2, R3, R4, R5 and R6, " substitution " refer to by it is selected from the group below it is one or more (such as 2,3 It is a, 4 etc.) substituent group replaced：Halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxies, halogenated C1-C6 It is alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo ,-CN, hydroxyl, amino, carboxyl, C6-C10 aryl, halogenated C6-C10 aryl, be selected from the group with 1-3 N, S and O heteroatomic 5-10 unit's heteroaryls, halogenated there is 1-3 to be selected From the heteroatomic 5-10 unit's heteroaryls of the following group N, S and O；

M is O, S, CR7R7 ' or NR8；Wherein, R7, R7 ', R8 be each independently hydrogen, halogen ,-CN, hydroxyl, substitution or Unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkoxyl, substituted or unsubstituted C3-C8 cycloalkyl；Wherein, institute " substitution " is stated to refer to be replaced by one or more (such as 2,3,4 etc.) substituent groups selected from the group below：Halogen, C1-C6 Alkyl, halogenated C1-C6 alkyl ,-CN, hydroxyl, amino, carboxyl；

X is NR9, carbonyl (- (CO) -), halogenated Asia C1-C4 alkyl (such as CF₂) or hydroxyl oxime (=N-OH)；Wherein, R9 for hydrogen, Substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C3-C8 cycloalkyl；Wherein, described " substitution " refers to by under One or more (such as 2,3,4 etc.) substituent group of group is replaced：Halogen, C1-C6 alkyl, halogenated C1-C6 alkane Base ,-CN, hydroxyl, amino, carboxyl；

Y is carbonyl (- (CO) -) or sulfonyl (- SO₂-)；

And meet the following conditions：X, Y cannot be simultaneously carbonyl, and Y cannot be sulfonyl when X is carbonyl；

Z is N or CR10；Wherein, R10 is each independently hydrogen, halogen ,-CN, hydroxyl, substituted or unsubstituted C1-C8 alkane Base, substituted or unsubstituted C1-C8 alkoxyl, substituted or unsubstituted C3-C10 cycloalkyl, it is substituted or unsubstituted have 1-3 A heteroatomic 3-10 membered heterocycloalkyls for being selected from the group N, S and O, substituted or unsubstituted C6-C10 aryl and substitution or not Substitution has the 1-3 heteroatomic 5-10 unit's heteroaryls for being selected from the group N, S and O；Wherein, described " substitution " refer to by selected from One or more (such as 2,3,4 etc.) substituent group of the following group is replaced：Halogen, C1-C6 alkyl, halogenated C1-C6 alkane Base, C1-C6 alkoxies, halogenated C1-C6 alkoxies, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo ,-CN, hydroxyl, Amino, C6-C10 aryl, halogenated C6-C10 aryl, has the 1-3 heteroatomic 5-10 for being selected from the group N, S and O members at carboxyl Heteroaryl, the halogenated heteroatomic 5-10 unit's heteroaryls for being selected from the group N, S and O with 1-3.

In another preferred example, R1, R2 are each independently hydrogen, substituted or unsubstituted C2-C8 alkyl, substitution or do not take The C3-C4 cycloalkyl in generation；Wherein, described " substitution " refers to by one or more (such as 2,3,4 etc.) selected from the group below Substituent group is replaced：Fluorine, chlorine, bromine, iodine.

In another preferred example, R1, R2 are each independently hydrogen, substituted or unsubstituted C2-C4 alkyl, substitution or do not take The C3-C4 cycloalkyl in generation；Wherein, described " substitution " refers to by one or more (such as 2,3,4 etc.) selected from the group below Substituent group is replaced：Fluorine, chlorine, bromine, iodine.

In another preferred example, R3 for hydrogen, fluorine, chlorine, bromine, iodine ,-CN, substituted or unsubstituted C1-C4 alkyl, substitution or Unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to by one or more (such as 2,3,4 selected from the group below Deng) substituent group replaced：Halogen, C1-C4 alkyl, halogenated C1-C4 alkyl ,-CN, hydroxyl.

In another preferred example, R3 is hydrogen, fluorine, chlorine, bromine, iodine.

In another preferred example, R4, R5 and R6 are each independently hydrogen, fluorine, chlorine, bromine, iodine ,-CN, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to by one or more selected from the group below (such as 2,3,4 etc.) substituent group is replaced：Fluorine, chlorine, bromine, iodine, C1-C4 alkyl, halogenated C1-C4 alkyl.

In another preferred example, R4, R5 and R6 are each independently hydrogen, fluorine, chlorine, bromine, iodine ,-CN, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to by one or more selected from the group below (such as 2,3,4 etc.) substituent group is replaced：Fluorine, chlorine, bromine, iodine.

In another preferred example, X NH, CF₂Or hydroxyl oxime (=N-OH).

In another preferred example, M NR8；Wherein, R8 is hydrogen, halogen ,-CN, hydroxyl, substituted or unsubstituted C1-C4 alkane Base, substituted or unsubstituted C1-C4 alkoxies, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refer to by One or more (such as 2,3,4 etc.) substituent group selected from the group below is replaced：Halogen, C1-C4 alkyl, halogenated C1- C4 alkyl ,-CN, hydroxyl, amino, carboxyl.

In another preferred example, Z is N or CR10；Wherein, R10 is each independently hydrogen, fluorine, chlorine, bromine, iodine, substitution or not Substituted C1-C4 alkyl, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to by one selected from the group below Or multiple (such as 2,3,4 etc.) substituent groups are replaced：Fluorine, chlorine, bromine, iodine, C1-C4 alkyl, halogenated C1-C4 alkyl.

In another preferred example, the compound is selected from the group：

Second aspect of the present invention provides the compound or its stereoisomer, cis-trans isomerism described in a kind of first aspect The preparation method of body or tautomer or its pharmaceutically acceptable salt, hydrate or solvate,

Method (a)：Shown formula A compounds are -1 compound represented of Formula VIII, the method includes the steps：

(a1) in atent solvent, compound III-1 is reacted with compound IV-1, so as to form compound V-1；

(a2) in atent solvent, reaction is hydrolyzed in compound V-1, so as to form compound VI-1；

(a3) in atent solvent, compound VI-1 is reacted with compound VII-1, so as to form compound VIII-1；

In various, R₁、R₂、R₃、R₄、R₅、R₆It is as defined above with Z；

Method (b)：Shown formula A compounds are -2 compound represented of Formula VIII, the method includes the steps：

(b1) in atent solvent, compound VII-2 and azanol are reacted, so as to form compound VIII-2；Respectively In formula, R₁、R₂、R₃、R₄、R₅、R₆It is as defined above with Z；

Method (c)：Shown formula A compounds are -2 compound represented of Formula IX, the method includes the steps：

(c1) in atent solvent, compound VII-2 is reacted with fluorination reagent, so as to form compound IX-2； In various, R₁、R₂、R₃、R₄、R₅、R₆It is as defined above with Z；

Method (d)：Shown formula A compounds are -3 compound represented of Formula IX, the method includes the steps：

(d1) in atent solvent, compound V-3 is reacted with sulfonic acid chloride, so as to form compound VI-3；

(d2) in atent solvent, by compound VI-3 and NR₁R₂It is reacted, so as to form compound VII-3；

(d3) in atent solvent, reaction is hydrolyzed in compound VII-3, so as to form compound VIII-3；

(d4) in atent solvent, compound VIII-3 is reacted with compound VII-1, so as to form compound IX-3；

In various, R₁、R₂、R₃、R₄、R₅And R₆It is as defined above.

In another preferred example, -2 compound of Formula VII can be prepared by -2 compound of Formulas I for starting material.Tool Body preparation method may include following steps：

In various, R₁、R₂、R₃、R₄、R₅、R₆It is as defined above with Z.

In another preferred example, -3 compound of Formula V can be prepared by -3 compound of Formulas I for starting material.

Specifically preparation method may include following steps：

In various, R₃It is as defined above.

Third aspect present invention provides the intermediate or its stereoisomer, cis-trans isomerism as shown in Formula V -1 or VI-1 Body or tautomer,

In various, R1, R2, R3 and Z are as defined above.

Fourth aspect present invention provides a kind of pharmaceutical composition, and comprising the compound described in (1) first aspect or it is vertical Body isomers, cis-trans-isomer or tautomer or its pharmaceutically acceptable salt, hydrate or solvate；(2) pharmacy Upper acceptable carrier.

In another preferred example, described pharmaceutical composition also is used to prevent and/or treat hepatitis type B virus comprising other The drug of infection.

In another preferred example, it is described other to may be selected from for preventing and/or treat hepatitis b virus infected drug The following group：The stimulant of immunomodulator (such as interferon-' alpha ' (IFN-α), glycol interferon-α) or innate immune system (such as 7 and/or 8 agonist of Toll-like receptor).

In another preferred example, it is described other to may be selected from for preventing and/or treat hepatitis b virus infected drug The following group：Tenofovir, tenofovir Chinese mugwort draw phenol amine (TAF), Lamivudine, adefovirdipivoxil, Entecavir and Sebivo.

Fifth aspect present invention provide compound described in first aspect or its stereoisomer, cis-trans-isomer or Pharmaceutical composition described in tautomer or its pharmaceutically acceptable salt, hydrate or solvate or fourth aspect Purposes is used to prepare prevention and/or treats hepatitis b virus infected drug.

Sixth aspect present invention provides a kind of method for treating hepatitis B, and first aspect is applied to individual in need The compound or its stereoisomer, cis-trans-isomer or tautomer or its pharmaceutically acceptable salt, hydration Pharmaceutical composition described in object or solvate or fourth aspect.

It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the invention and have in below (eg embodiment) It can be combined with each other between each technical characteristic of body description, so as to form new or preferred technical solution.As space is limited, exist This no longer tires out one by one states.

Specific embodiment

The present inventor is after extensive and in-depth study, it was found that a kind of to have excellent therapeutic effect to hepatitis B New compound.On this basis, inventor completes the present invention.

Definition

As used herein, the alkyl of term " alkyl " including linear chain or branch chain.Such as C1-C8 alkyl represents there is 1-8 The alkyl of the linear chain or branch chain of carbon atom, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl group, tertiary butyl etc..

As used herein, the alkenyl of term " alkenyl " including linear chain or branch chain.Such as C2-C6 alkenyls refer to 2-6 carbon The alkenyl of the linear chain or branch chain of atom, for example, vinyl, pi-allyl, 1- acrylic, isopropenyl, 1- cyclobutenyls, 2- cyclobutenyls, Or similar group.

As used herein, the alkynyl of term " alkynyl " including linear chain or branch chain.Such as C2-C6 alkynyls refer to 2-6 The alkynyl of the linear chain or branch chain of carbon atom, such as acetenyl, propinyl, butynyl or similar group.

As used herein, term " C3-C10 cycloalkyl " refers to the cycloalkyl with 3-10 carbon atom.It can be it is monocyclic, Such as cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl or similar group.Can also be bi-cyclic form, such as bridged ring or loop coil shape Formula.

As used herein, term " C1-C8 alkylamino radicals " refers to the amido replaced by C1-C8 alkyl, can be monosubstituted It is or disubstituted；For example, methylamino, ethylamino-, Propylamino, isopropylamine base, butylamine base, isobutyl amine, tert-butylamine base, dimethylamine Base, diethylin, dipropyl amido, diisopropylamino, dibutyl amino, di-iso-butylmanice base, two tert-butylamine bases etc..

As used herein, term " C1-C8 alkoxyl " refers to the alkoxy with the linear chain or branch chain of 1-8 carbon atom； For example, methoxyl group, ethyoxyl, propoxyl group, isopropoxy, butoxy, isobutoxy, tert-butoxy etc..

As used herein, " the heteroatomic 3-10 membered heterocycloalkyls that N, S and O are selected from the group with 1-3 " refer to 3-10 atom and cyclic group that wherein 1-3 atom is the heteroatomic saturation or fractional saturation that are selected from the group N, S and O Group.It can be monocyclic or bi-cyclic form, such as bridged ring or loop coil form.Specific example can be oxa- ring fourth Alkane, azetidine, tetrahydrochysene -2H- pyranoses, piperidyl, tetrahydrofuran base, morpholinyl and pyrrolidinyl etc..

As used herein, term " C6-C10 aryl " refers to there is the aryl of 6-10 carbon atom, for example, phenyl or naphthyl Deng similar group.

As used herein, term " the heteroatomic 5-10 unit's heteroaryls that N, S and O are selected from the group with 1-3 " refer to 5-10 atom and wherein 1-3 atom be the heteroatomic cyclic aromatic groups for being selected from the group N, S and O.It can be single Ring or condensed ring form.Specific example can be pyridyl group, pyridazinyl, pyrimidine radicals, pyrazinyl, triazine radical, pyrroles Base, pyrazolyl, imidazole radicals, (1,2,3)-triazolyl and (1,2,4)-triazolyl, tetrazole radical, furyl, thienyl, isoxazole Base, thiazolyl, oxazolyl etc..

Group of the present invention is " substituted or unsubstituted " unless stated otherwise, otherwise group of the invention Replaced by substituent group selected from the group below：Halogen, itrile group, nitro, hydroxyl, amino, C1-C6 alkyl-aminos, C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C1-C6 alkoxies, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyls, halogenated C2-C6 alkynyls, halogen For C1-C6 alkoxies, pi-allyl, benzyl, C6-C12 aryl, C1-C6 alkoxy -C 1-C6 alkyl, C1-C6 alkoxy-carbonyls, Carbobenzoxy, C2-C6 alkynyls-carbonyl, C2-C6 alkenyls-carbonyl, C3-C6 cycloalkyl-carbonyls, C1-C6 alkyl-sulfonyl bases etc..

As used herein, " halogen " or " halogen atom " refers to F, Cl, Br and I.More preferably, halogen or halogen atom are selected from F, Cl And Br." halogenated " refers to be replaced by the atom selected from F, Cl, Br and I.

Unless stated otherwise, structural formula described in the invention is intended to include all isomeric forms that (such as mapping is different Structure, diastereo-isomerism and geometric isomer (or rotamer))：Such as R, S configuration containing asymmetric center, double bond (Z), (E) isomers etc..Therefore, the single three-dimensional chemical isomer of the compounds of this invention or its enantiomter, diastereomeric are different The mixture of structure body or geometric isomer (or rotamer) belongs to the scope of the present invention.

As used herein, term " tautomer " represents that with different energy structural isomer can be more than low Energy barrier, so as to mutual inversion of phases.For example, proton tautomer (i.e. prototropic change) including by proton transfer carry out change, such as 1H- indazoles and 2H- indazoles.Valence tautomers include recombinating to carry out change by some bonding electrons.

As used herein, term " solvate " refers to that the compounds of this invention is coordinated to form special ratios with solvent molecule Complex.

As used herein, term " hydrate " refers to that the compounds of this invention carries out the complex of coordination formation with water.

Active constituent

As used herein, " the compounds of this invention " refers to formula (A) compound represented, and further includes and formula (A) compound Various crystalline forms, pharmaceutically acceptable salt, hydrate or solvate.

As used herein, " pharmaceutically acceptable salt " refer to the compounds of this invention with acid or alkali formed be suitable as medicine The salt of object.Pharmaceutically acceptable salt includes inorganic salts and organic salt.A kind of preferred salt is that the compounds of this invention is formed with acid Salt.The acid for suitably forming salt includes but is not limited to：The inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, Formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, hardship The organic acids such as sour, methanesulfonic acid, benzene methanesulfonic acid, benzene sulfonic acid；And the acidic amino acids such as aspartic acid, glutamic acid.

Preparation method

The compounds of this invention can be prepared according to method commonly used in the art.Also the scheme that can be given below according to the present invention Route is prepared using conventional synthesis condition (such as reaction temperature or reaction time etc.).Wherein, reaction temperature and reaction Time can be determined by those skilled in the art according to the popular response condition of the reaction.Reaction temperature can be -78 DEG C~reflux； Preferably, for -20 DEG C~reflux or 0~100 DEG C etc..Reaction time can be 0.1 hour~3 days or 0.1 hour~24 hours Or 0.1 hour~5 hours etc. the shorter time.

Scheme one：

(1) in atent solvent, at a certain temperature, compound I-1 is subjected to nitration reaction for a period of time, so as to be formed Compound II-1；

(2) in atent solvent, at a certain temperature, compound II-1 is subjected to reduction reaction for a period of time, so as to shape Into compound III-1；

(3) in atent solvent, at a certain temperature, when compound III-1 is carried out reacting one section with compound IV-1 Between, so as to form compound V-1；

(4) in atent solvent, at a certain temperature, reaction a period of time is hydrolyzed in compound V-1, so as to be formed Compound VI-1；With

(5) in atent solvent, at a certain temperature, when compound VI-1 is carried out reacting one section with compound VII-1 Between, so as to form compound VIII-1；

In various, R1, R2, R3, R4, R5, R6 and Z are as defined above.

Scheme two and scheme three：

(i) in atent solvent, at a certain temperature, reaction a period of time is hydrolyzed in compound I-2, so as to be formed Compound II-2；

(ii) in atent solvent, at a certain temperature, when compound II-2 is carried out reacting one section with aniline compound Between, so as to form compound III-2；

(iii) in atent solvent, at a certain temperature, compound III-2 and compound IV-2 are carried out reacting one section Time is so as to form compound V-2；

(iv) in atent solvent, at a certain temperature, reaction a period of time is hydrolyzed in compound V-2, so as to shape Into compound VI-2；

(v) in atent solvent, at a certain temperature, compound VI-2 and NR1R2 are carried out to react a period of time, so as to Form compound VII-2；With

(vi-1) in atent solvent, at a certain temperature, compound VII-2 and azanol are subjected to reaction a period of time, So as to form compound VIII-2；Or

(vi-2) in atent solvent, at a certain temperature, when compound VII-2 is carried out reacting one section with fluorination reagent Between, so as to form compound IX-2；

In various, R1, R2, R3, R4, R5, R6 and Z are as defined above.

In another preferred example, the fluorination reagent is the nucleophilic fluorination examination that carbonyl can be converted into difluoro methylene Agent, such as the conventional reagents such as DAST, BAST, Ishikawa reagent 1- fluorine pyridinium tetrafluoroborate salt.

Scheme four：

(a) in atent solvent, at a certain temperature, compound I-3 with methyl hydrazine react for a period of time, thus Form compound II-3；

(b) in atent solvent, at a certain temperature, by compound II-3 with carrying out bromination reaction for a period of time, so as to Form compound III-3；

(c) in atent solvent, at a certain temperature, compound III-3 is subjected to reaction a period of time, so as to formed Close object IV-3；

(d) in atent solvent, at a certain temperature, compound IV-3 is subjected to aminating reaction for a period of time, so as to shape Into compound V-3；

(e) in atent solvent, at a certain temperature, compound V-3 with sulfonic acid chloride react for a period of time, thus Form compound VI-3；

(f) in atent solvent, at a certain temperature, compound VI-3 and NR1R2 are carried out to react a period of time, so as to Form compound VII-3；

(g) in atent solvent, at a certain temperature, reaction a period of time is hydrolyzed in compound VII-3, so as to shape Into compound VIII-3；

(h) in atent solvent, at a certain temperature, compound VIII-3 and aniline are carried out to react a period of time, from And form compound IX-3；

In various, R1, R2, R3, R4, R5 and R6 are as defined above.

Intermediate

Include following compound suitable for the intermediate for preparing the compounds of this invention：

In various, R1, R2, R3 and Z are as defined above.

- 1 compound of Formula V can be prepared by -1 compound of Formulas I for starting material.Specifically preparation method may include Following steps：

In various, R1, R2, R3 and Z are as defined above.

- 1 compound of Formula IV can be prepared by -1 compound of Formulas I for starting material.Specifically preparation method can wrap Include following steps：

In various, R1, R2, R3 and Z are as defined above.

- 3 compound of Formula VII can be prepared by -1 compound of Formulas I for starting material.Specifically preparation method can wrap Include following steps：

In various, R1, R2 and R3 are as defined above.

- 3 compound of Formula VIII can be prepared by -3 compound of Formulas I for starting material.Specifically preparation method can Include the following steps：

In various, R1, R2 and R3 are as defined above.

Pharmaceutical composition and method of administration

Since the compounds of this invention has the inhibitory activity of excellent hepatitis type B virus (HBV), chemical combination of the present invention Object and its various crystal forms, pharmaceutically acceptable inorganic or organic salt, hydrate or solvate and contain chemical combination of the present invention Object can be used for preventing and/or treating and (stablize, mitigate or cure) hepatitis type B virus for the pharmaceutical composition of main active Infection or for prevent and/or treat (stablize, mitigate or cure) hepatitis type B virus relevant disease (for example, hepatitis B, into Malleability liver fibrosis, the inflammation for leading to hepatic sclerosis and necrosis, end-stage liver disease, ethyl liver cancer).

The pharmaceutical composition of the present invention includes the compounds of this invention in the range of safe and effective amount and can pharmaceutically receive Excipient or carrier.Wherein " safe and effective amount " refers to：The amount of compound is enough to be obviously improved the state of an illness, and is unlikely to generate Serious side effect.In general, pharmaceutical composition contains 1-2000mg the compounds of this invention/agent, more preferably, contain 10-200mg sheets Invention compound/agent.Preferably, described is " one " for a capsule or tablet.

" pharmaceutically acceptable carrier " refers to：One or more biocompatible solids or liquid filler or gelatinous mass, They are suitable for people's use and it is necessary to have enough purity and sufficiently low toxicity." compatibility " referred to herein as composition Middle each component energy and the compounds of this invention and they between mutually admix, and significantly reduce the drug effect of compound.Pharmaceutically Acceptable carrier part example has cellulose and its derivates (such as sodium carboxymethylcellulose, ethyl cellulose sodium, cellulose Acetic acid esters etc.), gelatin, talcum, kollag (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as soya-bean oil, sesame Oil, peanut oil, olive oil etc.), polyalcohol (such as propylene glycol, glycerine, mannitol, sorbierite), emulsifier (such as), profit Humectant (such as lauryl sodium sulfate), colorant, flavoring agent, stabilizer, antioxidant, preservative, apirogen water.

The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, and representative method of application includes (but being not limited to)：Oral, parenteral (intravenous, intramuscular or subcutaneous).

Include capsule, tablet, pill, powder and granule for the solid dosage forms of oral medication.In these solid formulations In type, reactive compound is mixed at least one conventional inert excipients (or carrier), such as sodium citrate or Dicalcium Phosphate or with Following compositions mix：(a) filler or bulking agent, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；(b) it bonds Agent, for example, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Arabic gum；(c) moisturizer, example Such as, glycerine；(d) disintegrant, for example, agar, calcium carbonate, potato starch or tapioca, alginic acid, certain composition silicates, And sodium carbonate；(e) retarding solvent, such as paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) wetting agent, such as spermaceti Alcohol and glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate, tristearin Sour magnesium, solid polyethylene glycol, lauryl sodium sulfate or its mixture.In capsule, tablet and pill, dosage form also may include Buffer.

Solid dosage forms such as tablet, sugar-pill, capsule, pill and granule can be used coating and shell material and prepare, such as casing and Other materials well known in the art.They may include opacifying agent, also, reactive compound or compound in this composition Release can discharge in certain part in the digestive tract in a delayed fashion.The example of adoptable embedding component is polymeric material And wax material.When necessary, reactive compound also can be with one or more formation microencapsulation forms in above-mentioned excipient.

Liquid formulation for oral administration includes pharmaceutically acceptable lotion, solution, suspension, syrup or tincture. In addition to active ingredient beyond the region of objective existence, liquid dosage form may include the inert diluent routinely used in this field, such as water or other solvents, increase Solvent and emulsifier, example know, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-BDO, dimethyl formyl The mixture of amine and oil, particularly cottonseed oil, peanut oil, maize germ, olive oil, castor oil and sesame oil or these substances Deng.

Other than these inert diluents, composition also may include auxiliary agent, such as wetting agent, emulsifier and suspending agent, sweet taste Agent, corrigent and fragrance.

In addition to active ingredient beyond the region of objective existence, suspension may include suspending agent, for example, ethoxylation isooctadecane alcohol, polyoxyethylene Sorbierite and the mixture of Isosorbide Dinitrate, microcrystalline cellulose, aluminium methoxide and agar or these substances etc..

For parenteral injection composition may include physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, Suspension or lotion with for re-dissolving into the aseptic powdery of sterile Injectable solution or dispersion liquid.It is suitable aqueous and Nonaqueous carrier, diluent, solvent or excipient include water, ethyl alcohol, polyalcohol and its suitable mixture.

The compounds of this invention can be administered alone or with other pharmaceutically acceptable compounds (such as anti-HBV agent) Administering drug combinations.

During administering drug combinations, described pharmaceutical composition further include with it is one or more (2 kinds, 3 kinds, 4 kinds, or more kind) other Pharmaceutically acceptable compound (such as anti-HBV agent).In other pharmaceutically acceptable compounds (such as anti-HBV agent) It is one or more (2 kinds, 3 kinds, 4 kinds, or more kind) can with the compound of the present invention simultaneously, separate or be sequentially used for preventing And/or treat HBV infection or HBV relevant diseases.

It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment during using pharmaceutical composition (such as people), wherein dosage is the effective dosage pharmaceutically thought during application, for the people of 60kg weight, day is to medicament Amount is usually 1~2000mg, preferably 20~500mg.Certainly, specific dosage is also contemplated that administration route, patient health situation etc. Factor, within the scope of these are all skilled practitioners technical ability.

Main advantages of the present invention include：

The compound of the present invention structure novel and there is excellent anti-hepatitis virus infection.

The compound of the present invention can effectively inhibit the assembling of hepatitis B nucleocapsid, so as to effectively inhibit hepatitis B.

The compound of the present invention is very low to the toxicity of normal cell.

The compounds of this invention and containing the compounds of this invention for main active pharmaceutical composition can be used for prevent And/or treatment is hepatitis b virus infected.

The compounds of this invention and containing the compounds of this invention for main active pharmaceutical composition can be used for prevent And/or treatment hepatitis type B virus relevant disease (for example, hepatitis B, progressivity liver fibrosis, the inflammation for leading to hepatic sclerosis and Necrosis, end-stage liver disease, ethyl liver cancer).

With reference to specific implementation, the present invention is further explained.It should be understood that these embodiments be merely to illustrate the present invention and It is not used in and limits the scope of the invention.Test method without specific conditions in the following example, usually according to normal condition, Or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise percentage and number are calculated by weight.The present invention is implemented Raw materials used or instrument in example, if not it illustrates, it is commercially available.

Embodiment 1：The synthesis of compound 8

Step 1：The synthesis of compound 2

Substrate 1 (1.0g) is dissolved in acetic anhydride (2mL), system temperature is down to 0 degree, then by fuming nitric aicd (0.5mL) Reaction system is added in, iodomethane (668mg) is then added in reaction system, reacts 3h at room temperature by 0 degree of reaction 3h.TLC detections are former The reaction was complete for material, water (20mL) is added in reaction system, ethyl acetate (3*30mL) extraction, organic phase is dry, is spin-dried for, crude product column Chromatograph (normal heptane：Ethyl acetate=1：5) product 2 (400mg), is obtained.MS (M+1=219).

Step 2：The synthesis of compound 3

Substrate 2 (0.4g) is dissolved in acetic acid (5mL), iron powder (1.8g) is added in into reaction system at room temperature, reacts at room temperature 3h, The reaction was complete for TLC detections raw material, after acetic acid is spin-dried for, adjusts the pH value of residue to 9-10, and ethyl acetate (3*25mL) extracts, Organic phase is dry, is spin-dried for, crude product column chromatography (normal heptane：Ethyl acetate=1：5) product 3 (200mg), is obtained.MS (M+1=189).

Step 3：The synthesis of compound 5

Substrate 3 (0.2g) is dissolved in acetonitrile (8mL), triethylamine (0.6g) is added in into reaction system at room temperature, then will be changed It closes object 4 (0.35g) and adds in reaction system, be heated to reflux 8h, the reaction was complete for TLC detections raw material, has new point generation, to reaction system In plus water (20mL), ethyl acetate (3*25mL) extraction, organic phase is dry, is spin-dried for, crude product column chromatography (normal heptane：Ethyl acetate =1：5) product 5 (80mg), is obtained.

¹H-NMR(CDCl₃,400MHz)δ:1.33 (d, J=6.8Hz, 3H), 3.85 (s, 3H), 3.86 (s, 3H), 3.95- 4.02 (m, 1H), 4.80 (d, J=9.2Hz, 1H), 6.19 (s, 1H), 6.90 (s, 1H)

Step 4：The synthesis of compound 6

Substrate 5 (80mg) is dissolved in tetrahydrofuran (2mL), in methanol (0.5mL) and water (0.5mL), then by hydroxide Lithium (0.21g) adds in reaction system, reacts 8h at room temperature, and TLC detection raw materials have a small amount of residue, there is new point generation.By water (20mL) adds in reaction system, with 1N hydrochloric acid tune pH value to 3-4, dichloromethane (3*25mL) extraction, organic phase anhydrous sodium sulfate It is dry, it is spin-dried for, crude product is through column chromatography (normal heptane：Ethyl acetate=1：5) yellow solid compound 6 (50mg) is detached to obtain.MS(M+ 1=350).

Step 5：The synthesis of compound 8

Substrate 6 (50mg) is dissolved in DMF (36mL), then by compound 7 (20mg), DIPEA (74mg) adds in reaction System, HATU (78mg) is added in lower reaction 5h, TLC the detection raw materials of about 30 degree of reaction system under 0 degree, and the reaction was complete, has new point to give birth to Into.Saturated salt solution 20mL is added in into reaction system, ethyl acetate (3*20mL) extraction, the drying of organic phase anhydrous sodium sulfate, rotation It is dry, crude product efficient liquid phase (normal heptane：Ethyl acetate=1：5) pink solid compound 8 (5mg) is detached to obtain.

¹H-NMR(CDCl₃,400MHz)δ:1.16 (d, J=6.8Hz, 3H), 3.76 (s, 3H), 3.92-3.99 (m, 1H), 7.52-7.57(m,1H),7.67(s,1H),7.94-7.98(m,1H),8.16-8.18(m,1H),8.49(br,1H)10.68 (s, 1H) .MS (M+1=468).

The synthesis of 2 compound 8a of embodiment

According to step 5, compound 7 need to only be replaced with compound 7a, other conditions are constant, through high performance liquid chromatography Detach to obtain target product 8a.MS (M+1=461).

The synthesis of 3 compound 8b of embodiment

According to step 5, compound 7 need to only be replaced with compound 7b, other conditions are constant, through high performance liquid chromatography Detach to obtain target product 8b.MS (M+1=457).

The synthesis of 4 compound 8c of embodiment

According to step 5, compound 7 need to only be replaced with compound 7c, other conditions are constant, through high performance liquid chromatography Detach to obtain target product 8c.MS (M+1=477).

The synthesis of 5 compound 8d of embodiment

According to step 5, compound 7 need to only be replaced with compound 7d, other conditions are constant, through high performance liquid chromatography Detach to obtain target product 8d.MS (M+1=495).

The synthesis of 6 compound 18/19 of embodiment

Step 11：The synthesis of compound 12

Substrate 11 (0.2g) is dissolved in tetrahydrofuran (2mL), in methanol (0.5mL) and water (0.5mL), then by hydroxide Lithium (0.24g) adds in reaction system, reacts 8h at room temperature, and TLC detection raw materials have a small amount of residue, there is new point generation.By water (20mL) adds in reaction system, with 1N hydrochloric acid tune pH value to 3-4, dichloromethane (3*25mL) extraction, organic phase anhydrous sodium sulfate It is dry, it is spin-dried for, crude product is through column chromatography (normal heptane：Ethyl acetate=1：5) compound as white solid 12 (0.16g) is detached to obtain.MS (M+1=160).

Step 12：The synthesis of compound 13

Substrate 12 (120mg) is dissolved in DMF (3mL), then by compound 12a (112mg), DIPEA (292mg) adds Entering reaction system, HATU (372mg) is added in into reaction system under 0 degree, about 30 degree lower reaction 5h, TLC detects raw materials, and the reaction was complete, There is new point generation.Saturated salt solution (20mL) is added in into reaction system, ethyl acetate (3*20mL) extraction, organic phase anhydrous slufuric acid Sodium is dried, and is spin-dried for, crude product is through column chromatography (normal heptane：Ethyl acetate=1：5) yellow solid compound 13 (90mg) is detached to obtain.MS (M+1=278).

Step 13：The synthesis of compound 15

Substrate 13 (90mg) is dissolved in DCM (3mL), system temperature is down to 0 degree, then by compound 14 (112mg) Dichloromethane solution add in reaction system, 30min is stirred under 0 degree, reaction then is added portionwise in alchlor (144mg) System, the reaction was complete for 0 degree of reaction 3h, TLC detection raw material, there is new point generation.Reaction solution is poured into ice water (10mL), dichloromethane Alkane (3*25mL) extracts, and organic phase wash with saturated salt solution 20mL, and anhydrous sodium sulfate is dried, and is spin-dried for, crude product through column chromatography (just Heptane：Ethyl acetate=1：8) yellow solid compound 15 (55mg) is detached to obtain.MS (M+1=378).

¹H-NMR(CDCl₃,400MHz)δ:1.38-1.42 (m, 3H), 4.02 (s, 3H), 4.35-4.41 (m, 1H), 7.19- 7.21(m,1H),7.67-7.72(m,1H),7.92(s,1H),8.06-8.08(m,1H),8.66(br,1H).

Step 14：The synthesis of compound 16

Substrate 15 (50mg) is dissolved in tetrahydrofuran (1mL) and ethyl alcohol (3mL), then will add in sodium hydroxide in system (16mg) aqueous solution, the reaction was complete for 30 degree of reaction 0.5h, TLC detection raw materials, there is new point generation.Water (20mL) is added in into reactant System, with 1N hydrochloric acid tune pH value to 3-4, dichloromethane (3*25mL) extraction, the drying of organic phase anhydrous sodium sulfate is spin-dried for, obtains yellow Solid chemical compound 16 (0.16g).

Step 15：The synthesis of compound 17

Substrate 16 (25mg) is dissolved in DMF (2mL), then by tert-butylamine (6.8mg), DIPEA (74mg) adds in reaction HATU (37mg) is added in reaction system under 0 degree by system, and about 30 degree lower reaction 5h, TLC detects raw materials, and the reaction was complete, there is new point Generation.Saturated salt solution (20mL) is added in into reaction system, ethyl acetate (3*20mL) extraction, organic phase anhydrous sodium sulfate is done It is dry, it is spin-dried for, crude product is through efficient liquid phase column (normal heptane：Ethyl acetate=1：5) light red solid compound 17 (5mg) is detached to obtain.

¹H-NMR(CDCl₃,400MHz)δ:1.41 (s, 9H), 4.00 (s, 3H), 7.19-7.23 (m, 1H), 7.69-7.77 (m, 1H), 8.04-8.06 (m, 1H), 8.54 (br, 1H) 8.68 (s, 1H) .Ms (ESI) m/z=403 (M-1).

Step 16：The synthesis of compound 18

Substrate 17 (15mg) is dissolved in ethyl alcohol (2mL), it, will be anti-then by the addition reaction system of hydroxylamine hydrochloride (13mg) Temperature is answered to rise to 60 degree of reaction 10h.Reaction solution is spin-dried for, after residual solids are dissolved with acetonitrile, efficient liquid phase reverse phase separation obtains Compound 18 (1.8mg).

¹H-NMR(CDCl₃,400MHz)δ:1.42 (s, 9H), 3.96 (s, 3H), 6.54-6.57 (m, 1H), 7.16-7.20 (m, 1H), 7.69-7.72 (m, 1H), 8.02 (s, 1H) 8.53 (d, J=8.0Hz, 1H) .Ms (ESI) m/z=418 (M-1)

Step 17：The synthesis of compound 19

Compound 17 (20mg) is dissolved in DCM (2mL), system temperature is then down to -78 degree, then by DAST (1mL) instills reaction system, the lower reaction 2h of -78 degree, and crude product is detached through performance liquid chromatographic column, obtains target product compound 19.MS (M+1=427).

The synthesis of 7 compound 18a of embodiment

According to step 11-15 prepare compound 17a, difference is to replace tert-butylamine with isopropylamine.

According to step 16, compound 17 need to only be replaced with compound 17a, other conditions are constant, through efficient liquid phase Chromatographic isolation obtains target product 18a.MS (M+1=406).

The synthesis of 8 compound 18b of embodiment

According to step 11-15 prepare compound 17b, difference is to replace tert-butylamine with ethamine.

According to step 16, compound 17 need to only be replaced with compound 17b, other conditions are constant, through efficient liquid phase Chromatographic isolation obtains target product 18b.MS (M+1=392).

The synthesis of 9 compound 18c of embodiment

According to step 11-15 prepare compound 17c, difference is with trifluoro isopropylamine (CH₃(CF₃)CHNH₂) instead of uncle Butylamine.

According to step 16, compound 17 need to only be replaced with compound 17c, other conditions are constant, through efficient liquid phase Chromatographic isolation obtains target product 18c.MS (M+1=460).

The synthesis of 10 compound 18d of embodiment

According to step 11-15 prepare compound 17d, difference is to replace tert-butylamine with cyclopropylamine.

According to step 16, compound 17 need to only be replaced with compound 17d, other conditions are constant, through efficient liquid phase Chromatographic isolation obtains target product 18d.MS (M+1=404).

The synthesis of 11 compound 19a of embodiment

According to the synthesis of step 17, compound 17 need to only be replaced with compound 17a, other conditions are constant, through height Effect liquid phase chromatogram detaches to obtain target product 19a.MS (M+1=413).

The synthesis of 13 compound 19b of embodiment

According to the synthesis of step 17, compound 17 need to only be replaced with compound 17b, other conditions are constant, through height Effect liquid phase chromatogram detaches to obtain target product 19b.MS (M+1=399).

The synthesis of 14 compound 19c of embodiment

According to the synthesis of step 17, compound 17 need to only be replaced with compound 17c, other conditions are constant, through height Effect liquid phase chromatogram detaches to obtain target product 19c.MS (M+1=467).

The synthesis of 15 compound 19d of embodiment

According to the synthesis of step 17, compound 17 need to only be replaced with compound 17d, other conditions are constant, through height Effect liquid phase chromatogram detaches to obtain target product 19d.MS (M+1=411).

The synthesis of 16 compound 20 of embodiment

Step 18：The synthesis of compound 22

Compounds methyl hydrazine (10g) is dissolved in methanol (20mL) and water (10mL), is cooled to 0 degree, is stirred after 15min so Compound 21 (12g) is added dropwise to reaction system afterwards, 70 degree of reaction 10h is heated to, system temperature is then returned back into room temperature, instead 12h is answered, is filtered, filter cake is washed with water (5mL), obtains 22 faint yellow solid 5g, MS (M+1=157).

Step 19：The synthesis of compound 23

Substrate 22 (5g) is dissolved in DMF (10mL), phosphorus oxychloride (10mL) is added in into reaction system at room temperature, is warming up to 100 degree, 5h is reacted, reaction system is poured into ice water, ethyl acetate (3*30mL) extraction, organic phase is dry, is spin-dried for, crude product column Chromatograph (normal heptane：Ethyl acetate=1：5) product 23 (2g), is obtained.MS (M+1=219).

Step 20：The synthesis of compound 24

Substrate 23 (2g) is dissolved in acetonitrile (10mL), NBS (4g) is added in into reaction system at room temperature, then by system temperature 60 degree are risen to, into reaction system plus water (20mL), ethyl acetate (3*25mL) extraction, organic phase drying are spin-dried for, crude product column layer Analyse (normal heptane：Ethyl acetate=1：5) product 5 (1g), is obtained.MS (M+1=299).

Step 21：The synthesis of compound 25

Substrate 24 (1g) is dissolved in dichloromethane (10mL), ammonium hydroxide (5mL) is then added in into reaction system, is reacted at room temperature Water (20mL) is added in reaction system by 2h, and with 1N hydrochloric acid tune pH value to 3-4, dichloromethane (3*25mL) extraction, organic phase is anhydrous Sodium sulphate is dried, and is spin-dried for, crude product is through column chromatography (normal heptane：Ethyl acetate=1：5) yellow solid compound 25 is detached to obtain (300mg).MS (M+1=234).

Step 22：The synthesis of compound 26

Substrate 25 (300mg) is dissolved in dichloromethane (3mL), then by sulfonic acid chloride (1mL), adds in reaction system, 40 Saturated salt solution 20mL is added in reaction system, ethyl acetate (3*20mL) extraction, organic phase anhydrous sodium sulfate by the lower reaction 5h of degree It is dry, it is spin-dried for, column chromatography (normal heptane：Ethyl acetate=1：3) yellow solid compound 26 (150mg) is detached to obtain.MS (M+1= 332)。

Step 23：The synthesis of compound 27

Substrate 26 (150mg) is dissolved in acetonitrile (5mL), triethylamine (0.1mL) is then added in into reaction system, then will Trifluoromethyl ethylamine hydrochloride (0.5g) adds in reaction system, and saturated salt solution 20mL is added in reactant by 80 degree of lower reaction 5h System, ethyl acetate (3*20mL) extraction, the drying of organic phase anhydrous sodium sulfate are spin-dried for, column chromatography (normal heptane：Ethyl acetate=1： 3) yellow solid compound 27 (120mg) is detached to obtain.MS (M+1=409).

Step 24：The synthesis of compound 28

Substrate 27 (120mg) is dissolved in tetrahydrofuran (2mL), in water (0.5mL) and methanol (0.5mL), then by a water It closes lithium hydroxide (100mg) and adds in reaction system, then 40 degree of lower reaction 8h, with 1N hydrochloric acid tune pH to 3-4, ethyl acetate (3* It 20mL) extracts, the drying of organic phase anhydrous sodium sulfate is spin-dried for, column chromatography (normal heptane：Ethyl acetate=1：3) detach yellow is consolidated Body compound 27 (80mg).MS (M+1=395).

Step 25：The synthesis of compound 20

By substrate 28 (80mg), compound 29 (50mg) and DIPEA (70mg) are dissolved in DMF (5mL), then by HATU (130mg) adds in reaction system, and then 40 degree lower reaction 8h, add water (20mL), and ethyl acetate (3*20mL) extracts, organic phase without Aqueous sodium persulfate is dried, and is spin-dried for, column chromatography (normal heptane：Ethyl acetate=1：3) yellow solid compound 20 (10mg) is detached to obtain.MS (M+1=506).

The synthesis of 17 compound 20b of embodiment

According to step 25, compound 29 is replaced with 29b, other are constant, column chromatography (normal heptane：Ethyl acetate=1：3) Detach to obtain yellow solid compound 20b (8mg).MS (M+1=522).

The synthesis of 18 compound 20c of embodiment

According to step 25, compound 29 is replaced with 29c, other are constant, column chromatography (normal heptane：Ethyl acetate=1：3) Detach to obtain yellow solid compound 20c (11mg).MS (M+1=502).

Biological examples -- anti-HBV activity is tested

Experiment one：Effect on hepatitics B virus in vitro nucleocapsid assembling activity test method

Main agents and raw material：

C150 albumen is expressed and is purified for Yao Ming Kants company；

FL is purchased from Thermo Fischer Scient Inc..

Protein fluorescence marks：

150 μ L 2%w/v skim milks are added in per hole to 96 orifice plates, are incubated at room temperature 2 hours.Skim milk is sopped up, is spent It is dried after ionized water cleaning, room temperature preservation.By C150 albumen (often 3 milligrams of pipe) 5ml Hitrap desalting column desalinations.To every pipe C150 albumen after desalination adds in 50mM20 μ l of FL fluorescent dyes are uniformly mixed, and 4 DEG C are protected from light overnight incubation.With The fluorescent dye that Sephadex G-25 gel filtrations removal is not combined with C150.The fluorescent label efficiency of C150 is calculated, formula is such as Under：

[FL]=A504/78,000M^-1；

[C150Bo]=(A280- [ FL]x 1300M^-1)/60,900M^-1；

Fluorescent label efficiency=[FL]/[C150Bo]；

Wherein,

[FL] represent fluorescent marker concentration；

[C150Bo] represents the concentration of fluorescent marker protein；

A504 represents the light absorption value of wavelength 504nM；

A280 represents the light absorption value of wavelength 280nM；

M^-1Represent the inverse of molar concentration.

Diluted chemical compound：

Compound stock solutions are diluted to 6mM with DMSO, then be diluted to 600 μM with 50mM HEPES, then use 10%DMSO/ Further 3 times of 50mM HEPES are serially diluted 8 concentration.

C150Bo is diluted to 2 μM with 50mM HEPES.Take the compound of 37.5 μ L C150Bo and the 2.5 each concentration of μ L Mixing in 96 hole reaction plates is added to, is incubated at room temperature 15 minutes.The 750mM NaCl/50mM HEPES of 10 μ l is taken to be added to reaction The final concentration of 150mM of Kong Zhong, NaCl.

0% albumen assembles control wells, adds in the final concentration of 0mM of the 50mM HEPES, NaCl of 10 μ L.

100% albumen assembles control wells, adds in the final concentration of 1M of 5M the NaCl/50mM HEPES, NaCl of 10 μ L.

DMSO final concentration of 0.5%, final concentration of 30 μM of compound highest, final concentration of 1.5 μM of C150Bo.Incubation at room temperature 1 hour.Measure fluorescence signal (exciting light 485nm；Emit light 535nm).

Data analysis

The assembling of % albumen=【1- (sample fluorescence value -1M NaCl fluorescent values)/(0M NaCl fluorescent value -1M NaCl fluorescence Value)】×100.

IC₅₀Value is calculated by prism softwares, and equation is as follows：

Y=Bottom+ (Top-Bottom)/(1+10^{((LogIC50-X)*HillSlope)})；

Wherein,

X represents the logarithm of concentration, and Y represents effect value, and Y is originated from bottom and is fitted to top with S types；

Bottom represents the bottom of curve；

Top represents that Top represents the top of curve；

HillSlope is represented：The absolute value of the greatest gradient of curve.

Experiment two：It is measured in the anti-hepatitis B activity of HepG2.2.15 cells

Main agents：

96 DNA Blood Kits (12) (Qiagen, article No. 51162) of QIAamp；

FastStart Universal Probe Master (Roche, article No. 04914058001)；

Cell-titer Glo detection reagents (Promega, article No. G7573).

Diluted chemical compound：External Anti-HBV effect experiment and equal 3 times of all compounds of cytotoxicity experiment are serially diluted, 8 Concentration.The final initial concentration of test-compound is 30 μM, and the final initial concentrations of reference compound GLS4 are 1 μM, DMSO final concentrations It is 0.5%.

Kind HepG2.2.15 cells (4 × 10⁴Cells/well) to 96 orifice plates, at 37 DEG C, 5%CO₂Overnight incubation.Second day, It adds in the fresh medium to culture hole of the compound containing various concentration.5th day, culture solution old in culture hole is absorbed, is added in The fresh medium of the compound containing various concentration.

8th day, the supernatant in culture hole is collected, for extracting HBV DNA, the qPCR detections HepG2.2.15 in supernatant HBV DNA contents in supernatant.After collecting supernatant, then supplemented medium and Cell-titer Glo reagents into culture hole, enzyme Mark the values of chemiluminescence that instrument detects each hole.

Active calculation formula is as follows:

Y=Bottom+ (Top-Bottom)/(1+10^{((LogIC50-X)*HillSlope)})；

Wherein,

Bottom represents the bottom of curve；

Top represents the top of curve；

Experiment three：Cytotoxicity assay

The cytotoxicity of untested compound is tested using HepG2 cells, by these cells in untested compound In the presence of be incubated 4 days.Cell viability is assessed using resazurin measure.

The result of above-mentioned experiment is shown in table 1.

1 compound activity data of table

All references mentioned in the present invention is incorporated herein by reference, independent just as each document It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can To be made various changes or modifications to the present invention, such equivalent forms equally fall within the model that the application the appended claims are limited It encloses.

Claims

1. it is a kind of as compound shown in formula A or its stereoisomer, cis-trans-isomer or tautomer or its pharmaceutically Acceptable salt, hydrate or solvate,

Wherein,

R1, R2 are each independently hydrogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, take In generation, unsubstituted there is 1-3 to be selected from the group the heteroatomic 3-10 membered heterocycloalkyls of N, S and O, substituted or unsubstituted C6- C10 aryl, the substituted or unsubstituted heteroatomic 5-10 unit's heteroaryls for being selected from the group N, S and O with 1-3；Or R1, R2 with The nitrogen-atoms being connected with them collectively forms the substituted or unsubstituted hetero atoms that there is 1 N and 0-3 to be selected from the group N, S and O 3-10 membered heterocycloalkyls；

R3 is hydrogen, halogen ,-CN, hydroxyl, amino, carboxyl ,-(C=O)-substituted or unsubstituted C1-C8 alkyl, replaces or does not take The C1-C8 alkyl in generation, substituted or unsubstituted C1-C8 alkylamino radicals, substituted or unsubstituted C1-C8 alkoxyl, substitution do not take The C3-C10 cycloalkyl in generation, the substituted or unsubstituted heteroatomic 3-10 circle heterocyclic rings alkane for being selected from the group N, S and O with 1-3 Base, substituted or unsubstituted C6-C10 aryl and substituted or unsubstituted there is 1-3 to be selected from the group the heteroatomic of N, S and O 5-10 unit's heteroaryls；

R4, R5 and R6 are the substituent group of any position on phenyl ring, are each independently hydrogen, halogen ,-CN, hydroxyl, amino, carboxylic Base ,-(C=O)-substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2- C6 alkenyls, substituted or unsubstituted C2-C6 alkynyls, substituted or unsubstituted C1-C8 alkylamino radicals, substituted or unsubstituted C1-C8 Alkoxy, substituted or unsubstituted C3-C10 cycloalkyl, the substituted or unsubstituted miscellaneous original for being selected from the group N, S and O with 1-3 The 3-10 membered heterocycloalkyls of son, substituted or unsubstituted C6-C10 aryl and substituted or unsubstituted there is 1-3 to be selected from the group N, the heteroatomic 5-10 unit's heteroaryls of S and O；

In R1, R2, R3, R4, R5 and R6, " substitution " refers to by one or more (such as 2,3,4 selected from the group below Deng) substituent group replaced：Halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxies, halogenated C1-C6 alkoxies, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo ,-CN, hydroxyl, amino, carboxyl, C6-C10 aryl, halogenated C6-C10 Aryl, halogenated has 1-3 is a to be selected from the group N, S at the heteroatomic 5-10 unit's heteroaryls for being selected from the group N, S and O with 1-3 With the heteroatomic 5-10 unit's heteroaryls of O；

M is O, S, CR7R7 ' or NR8；Wherein, R7, R7 ', R8 is each independently hydrogen, halogen ,-CN, hydroxyl, substitution or do not take The C1-C8 alkyl in generation, substituted or unsubstituted C1-C8 alkoxyl, substituted or unsubstituted C3-C8 cycloalkyl；Wherein, it is described " substitution " refers to be replaced by one or more (such as 2,3,4 etc.) substituent groups selected from the group below：Halogen, C1-C6 alkane Base, halogenated C1-C6 alkyl ,-CN, hydroxyl, amino, carboxyl；

X is NR9, carbonyl (- (CO) -), halogenated Asia C1-C4 alkyl (such as CF₂) or hydroxyl oxime (=N-OH)；Wherein, R9 is hydrogen, replaces Or unsubstituted C1-C8 alkyl, substituted or unsubstituted C3-C8 cycloalkyl；Wherein, described " substitution " refers to selected from the group below One or more (such as 2,3,4 etc.) substituent groups are replaced：Halogen, C1-C6 alkyl, halogenated C1-C6 alkyl ,-CN, Hydroxyl, amino, carboxyl；

Y is carbonyl (- (CO) -) or sulfonyl (- SO₂-)；

Z is N or CR10；Wherein, R10 be each independently hydrogen, halogen ,-CN, hydroxyl, substituted or unsubstituted C1-C8 alkyl, Substituted or unsubstituted C1-C8 alkoxyl, substituted or unsubstituted has 1-3 at substituted or unsubstituted C3-C10 cycloalkyl It is selected from the group heteroatomic 3-10 membered heterocycloalkyls, substituted or unsubstituted C6-C10 aryl and the substitution of N, S and O or does not take In generation, has the 1-3 heteroatomic 5-10 unit's heteroaryls for being selected from the group N, S and O；Wherein, described " substitution " refers to by under One or more (such as 2,3,4 etc.) substituent group of group is replaced：Halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxies, halogenated C1-C6 alkoxies, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo ,-CN, hydroxyl, ammonia Base, carboxyl, C6-C10 aryl, halogenated C6-C10 aryl, there is 1-3 to be selected from the group N, S and O heteroatomic 5-10 members it is miscellaneous Aryl, the halogenated heteroatomic 5-10 unit's heteroaryls for being selected from the group N, S and O with 1-3.

2. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy Upper acceptable salt, hydrate or solvate, which is characterized in that R1, R2 are each independently hydrogen, substituted or unsubstituted C2-C8 alkyl, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to by one or more selected from the group below (such as 2,3,4 etc.) substituent group is replaced：Fluorine, chlorine, bromine, iodine.

3. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy Upper acceptable salt, hydrate or solvate, which is characterized in that R3 is hydrogen, fluorine, chlorine, bromine, iodine ,-CN, substitution or unsubstituted C1-C4 alkyl, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to by one selected from the group below or more A (such as 2,3,4 etc.) substituent group is replaced：Halogen, C1-C4 alkyl, halogenated C1-C4 alkyl ,-CN, hydroxyl.

4. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy Upper acceptable salt, hydrate or solvate, which is characterized in that R4, R5 and R6 be each independently hydrogen, fluorine, chlorine, bromine, Iodine ,-CN, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C3-C4 cycloalkyl；Wherein, described " substitution " refers to Replaced by one or more (such as 2,3,4 etc.) substituent groups selected from the group below：Fluorine, chlorine, bromine, iodine, C1-C4 alkyl, Halogenated C1-C4 alkyl.

5. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy Upper acceptable salt, hydrate or solvate, which is characterized in that X NH, CF₂Or hydroxyl oxime (=N-OH).

6. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy Upper acceptable salt, hydrate or solvate, which is characterized in that the compound is selected from the group：

7. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy The preparation method of upper acceptable salt, hydrate or solvate, which is characterized in that

(b1) in atent solvent, compound VII-2 and azanol are reacted, so as to form compound VIII-2；In various, R₁、R₂、R₃、R₄、R₅、R₆It is as defined above with Z；

(c1) in atent solvent, compound VII-2 is reacted with fluorination reagent, so as to form compound IX-2；It is various In, R₁、R₂、R₃、R₄、R₅、R₆It is as defined above with Z；

In various, R₁、R₂、R₃、R₄、R₅And R₆It is as defined above.

8. intermediate or its stereoisomer, cis-trans-isomer or tautomer as shown in Formula V -1 or VI-1,

In various, R1, R2, R3 and Z are as defined above.

9. a kind of pharmaceutical composition, which is characterized in that include (1) compound as described in claim 1 or its alloisomerism Body, cis-trans-isomer or tautomer or its pharmaceutically acceptable salt, hydrate or solvate；(2) it can pharmaceutically connect The carrier received.

10. compound as described in claim 1 or its stereoisomer, cis-trans-isomer or tautomer or its pharmacy Upper acceptable salt, hydrate or the purposes of solvate or pharmaceutical composition as claimed in claim 9, which is characterized in that It is used to prepare prevention and/or treats hepatitis b virus infected drug.