WO2018133846A1 - Cyclothiourea compound and use thereof - Google Patents

Abstract

The present invention relates to a cyclothiourea compound and the use thereof. In particular, disclosed in the present invention are a compound having the structure as shown in formula (A) which can be used as an HBV replication inhibitor or a stereoisomer or a tautomer thereof, or a pharmaceutically acceptable salt, a hydrate or a solvate thereof. The present invention also relates to a pharmaceutical composition comprising the compound and the use thereof in treating hepatitis B.

Description

Cyclic thiourea compounds and uses thereof Technical field

The present invention pertains to the field of medicine, and in particular, to the use of cyclic thiourea compounds for the treatment of hepatitis B and uses thereof.

Background technique

Hepatitis B virus (HBV) is an enveloped, partially double-stranded DNA (dsDNA), hepatovirus DNA family (Hepadnaviridae) virus. Its genome contains four overlapping reading frames: the pronuclear/nuclear gene, the polymerase gene, the UM and S genes (which encode three envelope proteins), and the X gene. At the time of infection, the partially double-stranded DNA genome is transformed into a covalently closed circular DNA (cccDNA) in the host cell nucleus (open loop DNA, rcDNA) and the viral mRNA is transcribed. Once encapsidated, the pre-genomic RNA (pgRNA), which is also encoded by the core protein and Pol, is used as a template for reverse transcription, which regenerates this portion of the dsDNA genome (rcDNA) in the nucleocapsid.

The spread of hepatitis B virus is derived from exposure to infectious blood or body fluids, while viral DNA is detected in the saliva, tears, and urine of chronic carriers with high-priced DNA in serum. The choice of direct treatment is currently limited to interferon as well as the following antiviral drugs; tenofovir, lamivudine, adefovir, entecavir and telbivudine.

In addition, heteroaryldihydropyrimidines (HAPs) have been identified as a class of HBV inhibitors in tissue culture and animal models (Weber et al., Antiviral Res. 54:69-78) . Also disclosed are WO sulfonyl-aryl amide compounds which are resistant to HBV activity, as disclosed in WO 2013/006394 and WO 2013/096744.

However, these direct HBV antiviral drugs may encounter various problems such as toxicity, mutagenicity, lack of selectivity, poor efficacy, poor bioavailability, and difficulty in synthesis.

Therefore, there is a need in the art to develop HBV inhibitors having advantages such as high potency and lower toxicity.

Summary of the invention

It is an object of the present invention to provide a novel class of cyclic thiourea compounds useful as HBV inhibitors.

According to a first aspect of the present invention, there is provided a compound of formula A, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof,

among them,

R _{1 and} R ₂ are each independently selected from the group consisting of hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted having 1- 3 to 3 membered heterocycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, halogen or substituted or unsubstituted having 3 hetero atoms selected from the group consisting of N, S and O have 1-3 options from the group consisting of N, 5-10 membered heteroaryl hetero atoms of O and S, said R _1, the R _2, the substitution means one or more (e.g., two are selected from the group of 3 , 4, etc. substituted by a substituent: -OH, halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, =0, -O-;

X is CR ₁₁ R ₁₂ or -CR ₁₁ =CR ₁₂ -; wherein R ₁₁ and R ₁₂ are each independently selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or not Substituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted, having 1-3 selected from a 3-10 membered heterocycloalkyl group of a hetero atom of N, S and O, a substituted or unsubstituted C ₆ -C ₁₀ aryl group, substituted or unsubstituted, having 1-3 selected from the group consisting of N, S and a 5-10 membered heteroaryl, substituted or unsubstituted C _1-3 alkyl-R ₇ , -C(=O)OC _1-4 alkyl group of a hetero atom of O; wherein said R _{7 is} selected from the group consisting of : halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted 5-10 membered heteroaryl having 1-2 heteroatoms selected from the group consisting of N, S and O, having from 1 to 3 selected from a 3-7 membered heterocycloalkyl group of the following heteroatoms of N, S and O, -NR ₉ R ₁₀ , wherein said R ₉ and R ₁₀ are each independently selected from the group consisting of: H, C ₁ -C ₃ alkyl Halogenated C ₁ -C ₃ alkyl;

or,

The R ₁₁ and R ₁₂ together with the adjacent C atoms constitute a substituted or unsubstituted 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, wherein The substitution of the 3-7 membered heterocycloalkyl refers to the substitution of one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of -OH, halogen, methoxy, -O. -, -C(=O)OC _1-4 alkyl, benzyl, C _1-4 alkyl, halogenated C _1-4 alkyl,

Further, in the above R ₁₁ and R ₁₂ , the substitution means is substituted by one or more (for example, 2, 3, 4, etc.) substituents selected from the group consisting of -OH, halogen, C ₁ - C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, -OH substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -C(=O)OC _1-4 alkyl;

Y is a substituted or unsubstituted C ₁ -C ₇ alkylene group or a C ₂ -C ₇ alkenylene group, and in the Y, the substitution means one or more selected from the group consisting of (for example, 2, 3) Substituted by four, four, etc. substituents: C ₁ -C ₄ alkyl, halogen, -OH, preferably C ₁ -C ₄ alkyl or -OH;

Z is selected from the group consisting of NH, O or a bond;

Ring C is a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O, wherein said ring C is selected from the group consisting of Substituted by one or more (eg, 2, 3, 4, etc.) substituents: C ₁ -C ₃ alkyl (preferably methyl), C ₃ -C ₄ cycloalkyl, -CN or halogen ;

Ring B is a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 hetero atoms selected from the group consisting of N, S and O; In B, the substitution refers to being substituted with one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of C ₁ -C ₃ alkyl, C ₃ -C ₄ cycloalkyl , -CN or halogen;

R _a , R _b , R _c , R _d are substituents at any position on Ring B, each independently selected from the group consisting of H, halogen, —CN, hydroxy, amino, carboxy, —(C=O)- Substituted or unsubstituted C ₁ -C ₈ alkyl, substituted or unsubstituted C ₁ -C ₈ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkyne a substituted, unsubstituted or unsubstituted C ₁ -C ₈ alkylamino group, a substituted or unsubstituted C ₁ -C ₈ alkoxy group, a substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl group, substituted or unsubstituted 3 to 3 membered heterocycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, or 1-3 substituted or unsubstituted, selected from the group consisting of the following heteroatoms of N, S and O a 5-10 membered heteroaryl group selected from the group consisting of a hetero atom of N, S and O; in said R _a , R _b , R _c , R _d , said "substituted" means a group selected from the group consisting of Or a plurality of (eg, 2, 3, 4, etc.) substituents substituted: halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, Halogenated C ₁ -C ₆ alkoxy, C ₃ -C ₈ cycloalkyl, halogenated C ₃ -C ₈ cycloalkyl, oxo, -CN, hydroxy, amino, carboxy, C ₆ -C ₁₀ Aryl, Substituted C ₆ -C ₁₀ aryl group, having 1-3 heteroatoms selected from the group N, at 5-10 membered heteroaryl, S and O atoms in the aryl group, lower haloalkyl group having 1-3 heteroatoms selected from N, A 5-10 membered heteroaryl of a hetero atom of S and O.

In another preferred embodiment, R ₁ and R ₂ are each independently selected from the group consisting of hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted Or unsubstituted 3-10 membered heterocycloalkyl having 1 to 30 hetero atoms selected from the group consisting of N, S and O, substituted or unsubstituted C ₆ -C ₁₀ aryl, or substituted or unsubstituted a 5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S and O, wherein in R _{1 ,} R ₂ the substitution is replaced by one or more selected from the group consisting of Substituted by: -OH, halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -O-;

X is CR ₁₁ R ₁₂ or -CR ₁₁ =CR ₁₂ -; wherein R ₁₁ and R ₁₂ are each independently selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or not Substituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted, having 1-3 selected from a 3-10 membered heterocycloalkyl group of a hetero atom of N, S and O, a substituted or unsubstituted C ₆ -C ₁₀ aryl group, substituted or unsubstituted, having 1-3 selected from the group consisting of N, S and a 5-10 membered heteroaryl group of a hetero atom of O, a C _1-3 alkyl-R ₇ , or a -C(=O)OC _1-4 alkyl group; wherein said R _{7 is} selected from the group consisting of halogen, C _1- C ₃ alkyl, substituted or unsubstituted 5-10 membered heteroaryl having 1-2 heteroatoms selected from the group consisting of N, S and O, having 1-3 selected from the group consisting of N, a 3-7 membered heterocycloalkyl of a hetero atom of S and O, -NR ₉ R ₁₀ , wherein said R ₉ and R ₁₀ are each independently selected from the group consisting of: H, C ₁ -C ₃ alkyl, halogenated C ₁ -C ₃ alkyl;

or,

The R ₁₁ and R ₁₂ together with the adjacent C atoms constitute a substituted or unsubstituted 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, wherein The substitution of a 3-7 membered heterocycloalkyl group is substituted by one or more substituents selected from the group consisting of -OH, halogen, methoxy, -O-, -C(=O)OC _1-4 Alkyl, benzyl, C _1-4 alkyl, halogenated C _1-4 alkyl,

And, in R ₁₁ and R ₁₂ , the substitution means being substituted with one or more substituents selected from the group consisting of -OH, halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, -OH-substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -C (= O) OC _1-4 alkyl;

R _a , R _b , R _c and R _d are substituents at any position on the benzene ring, as defined above.

In another preferred embodiment, R ₁ is H, unsubstituted C ₁ -C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₁ -C substituted by -OH, =O, -O- or halogen ₁₀ alkyl.

In another preferred embodiment, R _{11 is} selected from the group consisting of H, substituted or unsubstituted C ₁ -C ₁₀ alkyl, -C(=O)OC _1-4 alkyl, C _1-3 alkyl-R _a substituted or unsubstituted C _2-4 alkynyl group, a 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, or a substituted or unsubstituted one having 1 Three 5-10 membered heteroaryl groups selected from the group consisting of the following heteroatoms of N, S and O.

In another preferred embodiment, R _{12 is} selected from the group consisting of H, substituted or unsubstituted C ₁ -C ₁₀ alkyl, preferably H, substituted or unsubstituted C ₁ -C ₆ alkyl, more preferably The ground is H or methyl.

In another preferred embodiment, ring C is a substituted or unsubstituted 5 or 6 membered heteroaryl group, said substitution being selected from one or more selected from the group consisting of (eg, 2, 3, 4, etc.) Substituted by a substituent: methyl, -CN or halogen.

In another preferred embodiment, Ring B is phenyl or a substituted or unsubstituted 6-membered heteroaryl group, preferably a phenyl or pyridyl group.

In another preferred embodiment, the R _a , R _b , R _c , R _d are each independently selected from the group consisting of H, halogen, —CHF ₂ , —CF ₂ —methyl, —CH ₂ F, —CF. ₃ , -OCF ₃ , -CN, -C ₃ -C ₄ cycloalkyl, or -C ₁ -C ₄ alkyl.

In another preferred embodiment, the ring C is

among them,

R ₄ is H, -C ₁ -C ₃ alkyl (preferably methyl), -C ₃ -C ₄ cycloalkyl;

R ₅ is H or halogen (preferably F);

R _{6 is} selected from H, methyl, -CN or halogen.

In another preferred embodiment, the Z is O or a bond.

In another preferred embodiment, the R ₇ is a substituted or unsubstituted 5-10 membered heteroaryl group having 1-2 heteroatoms selected from the group consisting of N, S and O.

In another preferred embodiment, the compound of formula A is selected from the group consisting of:

According to a second aspect of the present invention, there is provided a process for the preparation of a compound according to the first aspect of the invention, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, The compound of formula A is a compound of formula VII-1, and the method comprises the step (I):

or,

The compound of formula A shown is a compound of formula VIII-1, said method comprising step (II):

In the step (I) or (II), R _{2 ,} R _a , R _b , R _c , R _d , ring C, and ring B are as defined above, and m and n are each a positive integer of 1-5. ;

Wherein R _{3 is} selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkyne a substituted, unsubstituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted 3-10 membered heterocycloalkyl having 1 to 3 heteroatoms selected from the group consisting of N, S and O, substituted or Unsubstituted C ₆ -C ₁₀ aryl, or substituted or unsubstituted 5-10 membered heteroaryl, _1-3 alkyl group having 1-3 heteroatoms selected from the group consisting of N, S and O - R ₇ , —C(=O)OC _1-4 alkyl; wherein said R _{7 is} selected from the group consisting of halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted, and 1-2 selected from the group consisting of a 5-10 membered heteroaryl group of a hetero atom of N, S and O, a 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, -NR ₉ R ₁₀ Wherein R ₉ and R ₁₀ are each independently selected from the group consisting of: H, C ₁ -C ₃ alkyl, halogenated C ₁ -C ₃ alkyl.

According to a third aspect of the present invention, a pharmaceutical composition comprising the compound of the first aspect, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt or hydrate thereof, Or a solvate; (2) a pharmaceutically acceptable carrier.

In another preferred embodiment, the pharmaceutical composition further comprises other drugs for preventing and/or treating hepatitis B virus infection.

In another preferred embodiment, the other drug for preventing and/or treating hepatitis B virus infection may be selected from the group consisting of an immunomodulator (eg, interferon-α (IFN-α), PEGylation interference) Au-α) or a stimulant of the innate immune system (such as Toll-like receptor 7 and/or 8 agonists).

In another preferred embodiment, the other drug for preventing and/or treating hepatitis B virus infection may be selected from the group consisting of tenofovir, lamivudine, adefovir, entecavir, and telbiv. Set, or a combination thereof.

In a fourth aspect of the invention, there is provided a compound of the formula:

The fifth aspect of the invention provides a compound according to the first aspect of the invention, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a third aspect of the invention The use of the pharmaceutical composition for the preparation of a medicament for the prevention and/or treatment of hepatitis B virus infection.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

detailed description

The inventors have conducted extensive and intensive research and found a novel class of cyclic thiourea compounds which have an excellent therapeutic effect on hepatitis B. The compounds of the present invention have novel cores in structure, especially those having a cyclic thiourea, and therefore have not only excellent anti-HBV activity but also lower cytotoxicity (especially for liver cells). On this basis, the inventors completed the present invention.

definition

The term "alkyl" as used herein includes a straight or branched alkyl group. For example, C ₁ -C ₈ alkyl represents a straight or branched alkyl group having 1-8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl. Wait.

The term "alkenyl" as used herein, includes a straight or branched alkenyl group. For example, C ₂ -C ₆ alkenyl refers to a straight or branched alkenyl group having 2 to 6 carbon atoms, such as ethenyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2 a butenyl group, or a similar group.

The term "alkynyl", as used herein, includes a straight or branched alkynyl group. For example, C ₂ -C ₆ alkynyl refers to a straight or branched alkynyl group having 2 to 6 carbon atoms, such as an ethynyl group, a propynyl group, a butynyl group, or the like.

As used herein, the term "C ₃ -C ₁₀ cycloalkyl" means a cycloalkyl group having 3-10 carbon atoms. It may be a monocyclic ring such as a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, or the like. It may also be in the form of a double loop, such as a bridged or spiro ring.

As used herein, the term "C ₁ -C ₈ alkylamino" refers to an amine group substituted by a C ₁ -C ₈ alkyl group, which may be monosubstituted or disubstituted; for example, methylamino, ethylamino, Alanine, isopropylamino, butylamino, isobutylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, two Tert-butylamine and the like.

As used herein, the term "C ₁ -C ₈ alkoxy" refers to a straight or branched alkoxy group having from 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, iso Propyloxy, butoxy, isobutoxy, tert-butoxy and the like.

As used herein, the term "3-10 membered heterocycloalkyl having 1-3 heteroatoms selected from the group consisting of N, S and O" means having from 3 to 10 atoms and wherein 1-3 of the atoms are A saturated or partially saturated cyclic group selected from the group consisting of heteroatoms of N, S and O. It may be a single ring or a double ring form, such as a bridge ring or a spiro ring. Specific examples may be oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, pyrrolidinyl and the like.

As used herein, the term "C ₆ -C ₁₀ aryl" refers to an aryl group having 6 to 10 carbon atoms, for example, a phenyl or naphthyl group or the like.

As used herein, the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S and O" refers to having 5-10 atoms and wherein 1-3 atoms are selected from A cyclic aromatic group of the following group of heteroatoms of N, S and O. It may be a single ring or a fused ring. Specific examples may be pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2, 4)-Triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl and the like.

The groups of the present invention may be substituted with a substituent selected from the group consisting of halogen, nitrile group, nitro group, hydroxyl group, amino group, unless otherwise specified as "substituted or unsubstituted". , C ₁ -C ₆ alkyl-amino, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxy, halogenated C ₁ - C ₆ alkyl, halo C ₂ -C ₆ alkenyl, halo C ₂ -C ₆ alkynyl, halo C ₁ -C ₆ alkoxy, allyl, benzyl, C ₆ -C ₁₂ aryl , C ₁ -C ₆ alkoxy-C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy-carbonyl, phenoxycarbonyl, C ₂ -C ₆ alkynyl-carbonyl, C ₂ -C ₆ alkenyl a carbonyl group, a C ₃ -C ₆ cycloalkyl-carbonyl group, a C ₁ -C ₆ alkyl-sulfonyl group or the like.

As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from the group consisting of F, Cl and Br. "Halo" means substituted with an atom selected from the group consisting of F, Cl, Br, and I.

Unless otherwise stated, the structural formulae described herein are intended to include all isomeric forms (such as enantiomeric, diastereomeric, and geometric isomers (or conformational isomers): for example, containing asymmetry The central R, S configuration, the (Z) and (E) isomers of the double bond. Thus, a single stereochemical isomer of a compound of the invention, or a mixture of enantiomers, diastereomers or geometric isomers (or conformational isomers) thereof, is within the scope of the invention.

As used herein, the term "tautomer" means that structural isomers having different energies can exceed the low energy barrier and thereby transform each other. For example, proton tautomers (ie, proton shifts) include interconversions by proton transfer, such as 1H-carbazole and 2H-carbazole. Valence tautomers include interconversion through some bonding electron recombination.

As used herein, the term "solvate" refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a particular ratio.

As used herein, the term "hydrate" refers to a complex formed by the coordination of a compound of the invention with water.

Active ingredient

As used herein, "a compound of the invention" refers to a compound of formula (A), and also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula (A).

As used herein, "pharmaceutically acceptable salt" refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament. Pharmaceutically acceptable salts include inorganic and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Suitable acids for forming salts include, but are not limited to, mineral 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, Organic acids such as maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzoic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

Pharmaceutical composition and method of administration

Since the compound of the present invention has excellent inhibitory activity against hepatitis B virus (HBV), the compound of the present invention and various crystal forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and the present invention The pharmaceutical composition wherein the compound is the main active ingredient can be used for preventing and/or treating (stabilizing, alleviating or curing) hepatitis B virus infection or for preventing and/or treating (stabilizing, alleviating or curing) a hepatitis B virus-related disease ( For example, hepatitis B, progressive liver fibrosis, inflammation and necrosis leading to cirrhosis, terminal liver disease, ethyl liver cancer).

The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the invention and a pharmaceutically acceptable excipient or carrier. By "safe and effective amount" it is meant that the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of the compound of the invention per agent, more preferably from 10 to 200 mg of the compound of the invention per agent. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermixing with the compounds of the invention and between them without significantly reducing the potency of the compound. Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid). , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as

), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, parenteral (intravenous, intramuscular or subcutaneous).

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances.

In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds (e.g., anti-HBV agents).

When administered in combination, the pharmaceutical composition further comprises one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds (e.g., anti-HBV agents). One or more (2, 3, 4, or more) of the other pharmaceutically acceptable compound (eg, an anti-HBV agent) may be used simultaneously, separately or sequentially with the compound of the present invention. For the prevention and / or treatment of HBV infection or HBV related diseases.

When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually from 1 to 2000 mg, preferably from 20 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.

The compound of the invention:

The main advantages of the invention include:

1. The compound of the present invention is novel in structure and has an excellent anti-HBV infection effect.

2. The compounds of the invention are very toxic to normal cells.

3. The compound of the present invention and a pharmaceutical composition containing the compound of the present invention as a main active ingredient can be used for the prevention and/or treatment of hepatitis B virus infection.

4. The compound of the present invention and a pharmaceutical composition containing the compound of the present invention as a main active ingredient can be used for the prevention and/or treatment of hepatitis B virus-related diseases (for example, hepatitis B, progressive liver fibrosis, inflammation leading to cirrhosis, and Necrosis, terminal liver disease, ethyl liver cancer).

Terminology

All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise defined.

As used herein, when used in reference to a particular recited value, the term "about" means that the value can vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 and (eg, 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the terms "containing" or "including" may be open, semi-closed, and closed. In other words, the terms also include "consisting essentially of," or "consisting of."

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight and parts by weight.

The experimental materials and reagents used in the following examples are available from commercially available sources unless otherwise specified.

The following is the synthesis of 10 compounds:

Example 1: Synthesis of Compound 10a

step 1:

Compound 1 (1.5 g) was dissolved in acetic acid (20 mL), then iron powder (1.9 g) was added to the reaction system, the temperature was raised to 50 degrees for 2 h, and the reaction system was added with ethyl acetate (30 mL), and water (30 mL) was added. Ethyl acetate (3*20 mL) was extracted and dried over anhydrous sodium sulfate. ESI-MS, (M+H=189.04)

Step 2:

The chlorosulfonic acid isocyanate (200 mg) was dissolved in dichloromethane (5 mL), the temperature of the system was lowered to 0 °, then tert-butanol (105 mg) was added to the reaction system, stirred for 30 min, triethylamine (214 mg) and compound 2 (266 mg) was added to the reaction system at 0 °C. After the addition was completed, the mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was poured with water (20 mL), dichloromethane (3*20mL), and dried over anhydrous sodium sulfate Dry, crude column chromatography gave compound 3 (150 mg), ESI-MS, (M+H=368.06)

Step 3:

Compound 3 (100 mg) and 1,4-dibromobutane (58.8 mg) were dissolved in acetone (10 mL), and cesium carbonate (266 mg) was added to the reaction system, and reacted at 60 degrees for 24 hours, and water (15 mL) was added to the reaction system. Ethyl acetate (3*20 mL) was extracted, dried over anhydrous sodium sulfate, EtOAcjjjjjjjjjjjjjjj

Step 4:

Compound 4 (80 mg) was dissolved in tetrahydrofuran (2 mL), water (0.5 mL), methanol (0.5 mL), then lithium hydroxide monohydrate (80 mg) was added to the reaction system at room temperature, reacted at 40 °C for 5 h, then the system was The pH was adjusted to 3-4 with 1N EtOAc (EtOAc) (EtOAc (EtOAc)

Step 5:

Compound 5 (50 mg), triethylamine (70 mg), 4-fluoro-3-acetonitrile-1-aniline (22 mg) was dissolved in dichloromethane (3 mL), then the temperature was reduced to about 5 degrees, then TBTU (70 mg) was added to the reaction system, and the mixture was stirred at room temperature for 12 h, EtOAc (3 mL) -MS, (M+H=526.13)

Step 6:

Compound 6 (40 mg) was dissolved in dichloromethane (2 mL), then 4N HCl (1 mL) was added to the reaction system, and the reaction was carried out at 30 °C for 2 h, the reaction solution was spun dry, and the saturated sodium hydrogen carbonate solution was adjusted to pH 7-8, acetic acid. Ethyl acetate (3*10 mL) was extracted, dried over anhydrous sodium sulfate, and then evaporated to dryness.

Example 2: Synthesis of Compound 10b

The preparation of the compound 6b was carried out in the same manner as in the step 1-5 of Example 1, except that in step 3, 1,3-dibromopropane was used instead of 1,4-dibromobutane.

According to the step 6 of Example 1, only the compound 6b was used instead of the compound 6, and the other conditions were unchanged, and the objective product 10b (11 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 1:1).

Example 3: Synthesis of Compound 10c

The preparation of Compound 6c was carried out in the same manner as in Step 1-5 of Example 1, except that 1,5-dibromopentane was used in place of 1,4-dibromobutane in Step 3.

According to the step 6 of Example 1, only the compound 6c was used instead of the compound 6, and the other conditions were unchanged, and the objective product 10c (6 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 4: Synthesis of Compound 10d

The preparation of the compound 6d was carried out in the same manner as in the step 1-5 of Example 1, except that in step 3, 2,5-dibromopentane was used instead of 1,4-dibromobutane.

According to the step 6 of the first embodiment, the compound 6d was used instead of the compound 6, and the other conditions were unchanged, and the objective product was obtained by column chromatography (n-heptane: ethyl acetate = 2:1) 10d (7mg).

Example 5: Synthesis of Compound 10e

The preparation of the compound 6e was carried out in the same manner as in the step 1-5 of Example 1, except that in Step 3, 2-difluoromethyl-5-bromopropane was used instead of 1,4-dibromobutane.

According to the step 6 of Example 1, it was only necessary to replace the compound 6 with the compound 6e, and the other conditions were unchanged, and the objective product 10e (11 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 6: Synthesis of Compound 10f

The preparation of the compound 6f was carried out in the same manner as in the step 1-5 of Example 1, except that in Step 3, 2-methylisoxazole benzyl-2,5-dibromohexane was used instead of 1,4-dibromobutane.

According to the step 6 of Example 1, it was only necessary to replace the compound 6 with the compound 6f, and the other conditions were unchanged, and the objective product 10f (8 mg) was obtained by column chromatography (n-heptane: ethyl acetate = 1:1).

Example 7: Synthesis of Compound 10g

The preparation of the compound 6g is carried out in the same manner as in the step 1-5 of the embodiment 1, except that the 1-isopropyl-1,4-dibromohexane is substituted for the 1,4-dibromobutane in the step 3, and the step 5 is used. 3,4-Difluoro-aniline instead of 4-fluoro-3-cyanoaniline.

According to the step 6 of Example 1, only the compound 6g was used instead of the compound 6, and the other conditions were unchanged, and the objective product (10 g (4 mg)) was obtained by column chromatography (n-heptane: ethyl acetate = 1:1).

Example 8: Synthesis of Compound 10h

The preparation of the compound 6h is carried out by referring to the steps 1-5 of the first embodiment, except that the 1-isopropyl-1,4-dibromohexane is substituted for the 1,4-dibromobutane in the step 3, and the step 5 is used. 4-Fluoro-3-difluoromethylaniline instead of 4-fluoro-3-cyanoaniline.

According to the step 6 of Example 1, the compound 6h was used instead of the compound 6, and the other conditions were unchanged, and the objective product was subjected to column chromatography (n-heptane: ethyl acetate = 1:1) for 10 h (9 mg).

Example 9: Synthesis of Compound 20a

Step 11:

Allyl bromide (1.4g) was added dropwise to a solution of compound 2 (2g) and potassium carbonate (2.3g) in acetonitrile (30mL), the temperature of the system was raised to 85 degrees, reacted for 1h, water was added to the reaction system, acetic acid Ethyl acetate (3*20 mL) was extracted and dried over anhydrous sodium sulfate. The organic phase was evaporated to dryness.

Step 12:

The chlorosulfonic acid isocyanate (2 g) was dissolved in dichloromethane (40 mL), the temperature of the system was lowered to 0 °, then tert-butanol (1.3 g) was added to the reaction system, stirred for 30 min, triethylamine (3 g) and compound 12(2.0g) was added to the reaction system at 0 degree. After the addition was completed, the temperature was raised to room temperature and the reaction was stirred for 2 h. The reaction system was added with water (50 mL), dichloromethane (3*50 mL), and dried over anhydrous sodium sulfate The phase was dried and the crude column was purified to afford compound 13 (1.4 g), ESI-MS, (M+H= 408.10)

Step 13:

Compound 13 (1.2 g) and allyl bromide (1.1 g) were dissolved in acetonitrile, then cesium carbonate was added to the reaction system, the temperature was raised to 80 degrees, and the reaction was carried out for 2 hours, and water (20 mL) was added to the reaction system, ethyl acetate ( 3*20 mL) was extracted, dried over anhydrous sodium sulfate and the organic phase was dried to give compound 14 (1 g) ESI-MS (M+H=448.12)

Step 14:

The compound 14 (400 mg) was dissolved in dichloromethane (400 mL), then the catalyst CatB (42 mg) was added to the reaction system, and the reaction was carried out at 30 degrees for 12 hours. The reaction system was added to silica gel to dry to a white powder, and the compound was obtained by column chromatography. 15 (130 mg) ESI-MS, (M+H=420.09)

Step 15:

The compound 15 (80 mg) was dissolved in tetrahydrofuran (2 mL), water (0.5 mL), methanol (0.5 mL), then lithium hydroxide monohydrate (80 mg) was added to the reaction system at room temperature, reacted at 40 ° C for 5 h, then the system was The pH was adjusted to 3-4 with 1N EtOAc (EtOAc) (EtOAc (EtOAc)

Step 16:

Compound 16 (40 mg), triethylamine (60 mg), 4-fluoro-3-acetonitrile-1-aniline (22 mg) was dissolved in dichloromethane (3 mL), then the temperature was reduced to about 5 degrees, then TBTU (70 mg) was added to the reaction system, and the mixture was stirred at room temperature for 12 h, EtOAc (3 mL) -MS, (M+H=524.11)

Step 17:

The compound 17 (40 mg) was dissolved in dichloromethane (2 mL) and then 4N HCl (1 mL) was added to the reaction system, and the reaction was carried out at 30 °C for 2 h, the reaction solution was spun dry, and the saturated sodium hydrogen carbonate solution was adjusted to pH 7-8, acetic acid. Ethyl ester (3*10 mL) was extracted, dried over anhydrous sodium sulfate, and then evaporated to dryness.

Example 10: Synthesis of Compound 20b

The procedure for the preparation of compound 17b is as described in Steps 11-16 of Example 9, except that in step 13, 4-bromo-1-butene was used in place of propylene bromide.

According to the step 17 of Example 9, the compound 17b was replaced with the compound 17b, and the other conditions were unchanged, and the objective product 20b (6 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 11: Synthesis of Compound 20c

The procedure for the preparation of compound 17c is as described in Steps 11-16 of Example 9, except that in step 13, 3-bromo-1-butene was used in place of propylene bromide.

According to the step 17 of Example 9, only the compound 17c was used instead of the compound 17 and the other conditions were unchanged, and the objective product 20c (13 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 12: Synthesis of Compound 20d

The preparation of the compound 17d is carried out by referring to the step 11-16 of Example 9, except that in Step 13, 4-methylisoxazole benzyl-1-pentene was used instead of propylene bromide.

According to the step 17 of Example 9, the compound 17d was replaced with the compound 17d, and the other conditions were unchanged, and the objective product was 20d (22 mg) by column chromatography (n-heptane: ethyl acetate = 2:1).

Example 13: Synthesis of Compound 20e

The preparation of the compound 17e is carried out by referring to the steps 11-16 of Example 9, except that in step 13, 1-isopropyl-1-propene is used instead of propylene bromide, and in step 16, 3,4-difluoro-aniline is used instead of 4- Fluoro-3-cyanoaniline.

According to the step 17 of Example 9, the compound 17e was replaced with the compound 17e, and the other conditions were unchanged, and the objective product 20e (14 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 1:1).

Example 14: Synthesis of Compound 20f

The preparation of compound 17f is carried out in the same manner as in the step 11-16 of Example 9, except that in step 13, 1-isopropyl-1-propene is used in place of propylene bromide, and in step 16, 4-fluoro-3-difluoromethyl group is used. Aniline replaces 4-fluoro-3-cyanoaniline.

According to the step 17 of Example 9, the compound 17f was used instead of the compound 17 and the other conditions were unchanged, and the objective product 20f (8 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 1:1).

Example 15: Synthesis of Compound 30a

Step 21:

Compound 21 (1.5 g) was dissolved in acetic acid (20 mL), then iron powder (1.9 g) was added to the reaction system, the temperature was raised to 50 degrees for 2 h, and the reaction system was added with ethyl acetate (30 mL), and water (30 mL) was added. Ethyl acetate (3*20 mL) was extracted and dried over anhydrous sodium sulfate. ESI-MS, (M+H=190.03)

Step 22:

The chlorosulfonic acid isocyanate (200 mg) was dissolved in dichloromethane (5 mL), the temperature of the system was lowered to 0 °, then tert-butanol (105 mg) was added to the reaction system, stirred for 30 min, triethylamine (214 mg) and compound 22 (266 mg) was added to the reaction system at 0 °C. After the addition was completed, the mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was poured with water (20 mL), dichloromethane (3*20mL), and dried over anhydrous sodium sulfate Dry, crude column chromatography gave compound 23 (150 mg), ESI-MS, (M+H=369.06)

Step 23:

Compound 23 (100 mg) and 1,4-dibromobutane (58.8 mg) were dissolved in acetone (10 mL), and cesium carbonate (266 mg) was added to the reaction system, and reacted at 60 degrees for 24 hours, and water (15 mL) was added to the reaction system. Ethyl acetate (3*20 mL) was evaporated. EtOAcjjjjjjjjjj

Step 24:

The compound 24 (80 mg) was dissolved in tetrahydrofuran (2 mL), water (0.5 mL), methanol (0.5 mL), then lithium hydroxide monohydrate (80 mg) was added to the reaction system at room temperature, reacted at 40 °C for 5 h, then the system was The pH was adjusted to 3-4 with 1N EtOAc (EtOAc) (EtOAc (EtOAc)

Step 25:

Compound 25 (50 mg), triethylamine (70 mg), 4-fluoro-3-acetonitrile-1-aniline (22 mg) was dissolved in dichloromethane (3 mL), then the temperature was reduced to about 5 degrees, then TBTU (70 mg) was added to the reaction system, and the mixture was stirred at room temperature for 12 h, EtOAc (3 mL) (EtOAc) -MS, (M+H=526.13)

Step 26:

The compound 26 (40 mg) was dissolved in dichloromethane (2 mL) and then 4N HCl (1 mL) was added to the reaction system, and the reaction was carried out at 30 °C for 2 h, the reaction solution was spun dry, and the saturated sodium hydrogen carbonate solution was adjusted to pH 7-8, acetic acid. Ethyl acetate (3*10 mL) was extracted, dried over anhydrous sodium sulfate, and then evaporated to dryness.

Example 16: Synthesis of Compound 30b

The procedure for the preparation of compound 26b is as described in Steps 21-25 of Example 15, except that in step 23, 2,4-dibromopentane is used in place of 1,4-dibromobutane.

According to the step 26 of Example 15, the compound 26b was used instead of the compound 26, and the other conditions were unchanged, and the objective product 30b (11 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 1:1).

Example 17: Synthesis of Compound 30c

The procedure for the preparation of compound 26c is as described in Steps 21-25 of Example 15, except that in Step 23, 2-methylisoxazole benzyl-2,5-dibromohexane was used in place of 1,4-dibromobutane.

According to the step 26 of Example 15, the compound 26c was used instead of the compound 26, and the other conditions were unchanged, and the objective product 30c (8 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 18: Synthesis of 30d

The preparation of the compound 26d was carried out by referring to the step 21-25 of Example 15, except that in Step 23, 1-isopropyl-1,4-dibromohexane was used instead of 1,4-dibromobutane. In step 25, 3,4-difluoro-aniline is used in place of 4-fluoro-3-cyanoaniline. According to the step 26 of Example 15, the compound 26d was used instead of the compound 26, and the other conditions were unchanged, and the objective product was 30d (9 mg) by column chromatography (n-heptane: ethyl acetate = 2:1).

Example 19: Synthesis of Compound 30e

The preparation of compound 26e is carried out by referring to Steps 21-25 of Example 15 except that in Step 23, 1-isopropyl-1,4-dibromohexane was used instead of 1,4-dibromobutane. In step 25, 4-fluoro-3-cyanoaniline is replaced with 3-difluoromethyl-4-fluoro-aniline. According to the step 26 of Example 15, the compound 26e was used instead of the compound 26, and the other conditions were unchanged, and the objective product 30e (11 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 20: Synthesis of Compound 40a

Step 31:

Allyl bromide (1.4g) was added dropwise to a solution of compound 31 (2g) and potassium carbonate (2.3g) in acetonitrile (30mL), the temperature of the system was raised to 85 degrees, the reaction was carried out for 1h, water was added to the reaction system, acetic acid Ethyl acetate (3*20 mL) was extracted, dried over anhydrous sodium sulfate, and then evaporated.

Step 32:

The chlorosulfonic acid isocyanate (2 g) was dissolved in dichloromethane (40 mL), the temperature of the system was lowered to 0 °, then tert-butanol (1.3 g) was added to the reaction system, stirred for 30 min, triethylamine (3 g) and compound 32 (2.0 g) was added to the reaction system at 0 °C. After the addition was completed, the mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was poured with water (50mL), dichloromethane (3*50mL), and dried over anhydrous sodium sulfate The phase was dried and the crude column was purified to afford compound 33 (1.4 g), ESI-MS, (M+H=409.11)

Step 33:

Compound 33 (1.2 g) and allyl bromide (1.1 g) were dissolved in acetonitrile, then cesium carbonate was added to the reaction system, the temperature was raised to 80 degrees, and the reaction was carried out for 2 hours, and water (20 mL) was added to the reaction system, ethyl acetate ( 3*20 mL) was extracted, dried over anhydrous sodium sulfate, and the organic phase was dried to give compound 34 (1 g) ESI-MS (M+H=449.13)

Step 34:

The compound 34 (400 mg) was dissolved in dichloromethane (400 mL), then the catalyst CatB (42 mg) was added to the reaction system, and the reaction was carried out at 30 degrees for 12 hours. The reaction system was added to silica gel to spin to a white powder, and the compound was obtained by column chromatography. 35 (130 mg) ESI-MS, (M+H=421.10)

Step 35:

The compound 35 (80 mg) was dissolved in tetrahydrofuran (2 mL), water (0.5 mL), methanol (0.5 mL), and then lithium hydroxide monohydrate (80 mg) was added to the reaction system at room temperature, and reacted at 40 ° C for 5 h, then the system was The pH was adjusted to 3-4 with 1N EtOAc (EtOAc) (EtOAc (EtOAc)

Step 36:

Compound 36 (40 mg), triethylamine (60 mg), 4-fluoro-3-acetonitrile-1-aniline (22 mg) was dissolved in dichloromethane (3 mL), then the temperature was reduced to about 5 degrees, then TBTU (70 mg) was added to the reaction system, and the mixture was stirred at room temperature for 12 h, EtOAc (3 mL) -MS, (M+H=525.11)

Step 37:

The compound 37 (40 mg) was dissolved in dichloromethane (2 mL) and then 4N HCl (1 mL) was added to the reaction system, and the reaction was carried out at 30 °C for 2 h, the reaction solution was spun dry, and the saturated sodium hydrogen carbonate solution was adjusted to pH 7-8. Ethyl ester (3*10 mL) was extracted, dried over anhydrous sodium sulfate, and then evaporated to dryness.

Example 21: Synthesis of Compound 40b

The procedure for the preparation of compound 37b is as described in steps 31-36 of Example 20, except that in step 32, 3-bromo-1-butene is used in place of propylene bromide.

According to the step 37 of Example 20, the compound 37b was used instead of the compound 37, and the other conditions were unchanged, and the objective product 40b (6 mg) was subjected to column chromatography (n-heptane: ethyl acetate = 2:1).

Example 22: Synthesis of Compound 40c

The preparation of compound 37c is carried out in accordance with steps 31-36 of Example 20, except that 4-methyl isoxazole benzyl-1-pentene is substituted for propylene bromide in step 32 according to step 37 of Example 20, using only The title compound 40c (13 mg) was obtained by column chromatography (n-heptane: ethyl acetate = 2:1).

Example 23: Synthesis of Compound 40d

The preparation of compound 37d is carried out by referring to steps 31-36 of Example 20, except that in step 32, 1-isopropyl-1-propene is used in place of propylene bromide, and in step 35, 3,4-difluoro-aniline is substituted for 4- Fluoro-3-cyanoaniline.

According to the step 37 of Example 20, the compound 37d was used instead of the compound 37, and the other conditions were unchanged, and the objective product was 40d (12 mg) by column chromatography (n-heptane: ethyl acetate = 2:1).

Example 24: Synthesis of Compound 40e

The preparation of compound 37e is carried out by referring to steps 31-36 of Example 20, except that in step 32, 1-isopropyl-1-propene is used in place of propylene bromide, and in step 35, 3-difluoromethyl-4-fluoro- is used. Aniline replaces 4-fluoro-3-cyanoaniline.

According to the step 37 of Example 20, the compound 37e was used instead of the compound 37, and the other conditions were unchanged, and the objective product 40e (13 mg) was obtained by column chromatography (n-heptane: ethyl acetate = 2:1).

Biological Example--Anti-HBV Activity Experiment

Experiment 1: In vitro anti-HBV nucleocapsid assembly activity test method

Main reagents and raw materials:

C150 protein is expressed and purified by WuXi PharmaTech;

Purchased from Thermo Fisher Scientific.

Protein fluorescent label:

150 μL of 2% w/v skim milk was added to each well of a 96-well plate and incubated for 2 hours at room temperature. Aspirate skim milk, wash it with deionized water, dry it, and store at room temperature. C150 protein (3 mg per tube) was desalted using a 5 ml Hitrap desalting column. Add 50 mM to the C150 protein after desalting each tube

20 μl of the fluorescent dye was mixed well and incubated overnight at 4 ° C in the dark. The fluorescent dye not bound to C150 was removed by filtration with Sephadex G-25 gel. Calculate the fluorescence labeling efficiency of C150, the formula is as follows:

among them,

Indicates the concentration of the fluorescent label;

[C150Bo] represents the concentration of a fluorescently labeled protein;

A504 represents an absorbance value of a wavelength of 504 nM;

A280 represents an absorption value of a wavelength of 280 nM;

M ^-1 represents the reciprocal of the molar concentration.

Compound dilution:

The mother liquor of the compound was diluted to 6 mM with DMSO, diluted to 600 μM with 50 mM HEPES, and then further diluted 8 times with 10% DMSO/50 mM HEPES.

C150Bo was diluted to 2 μM with 50 mM HEPES. 37.5 μL of C150Bo and 2.5 μL of each concentration of the compound were added to a 96-well reaction plate and mixed, and incubated at room temperature for 15 minutes. 10 μl of 750 mM NaCl/50 mM HEPES was added to the reaction well, and the final concentration of NaCl was 150 mM.

Control wells were assembled with 0% protein, 10 μL of 50 mM HEPES was added, and the final concentration of NaCl was 0 mM.

Control wells were assembled with 100% protein and 10 μL of 5 M NaCl/50 mM HEPES was added with a final concentration of 1 M NaCl.

The final concentration of DMSO was 0.5%, the maximum final concentration of the compound was 30 μM, and the final concentration of C150Bo was 1.5 μM. Incubate for 1 hour at room temperature. The fluorescence signal (excitation light 485 nm; emission light 535 nm) was measured.

data analysis

% protein assembly = [1 - (sample fluorescence value - 1 M NaCl fluorescence value) / (0 M NaCl fluorescence value - 1 M NaCl fluorescence value)] × 100.

The IC ₅₀ value is calculated by the prism software and the equation is as follows:

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

Where X represents the logarithm of the concentration, Y represents the effect value, and Y is fitted from the bottom to the top with an S-shape;

Bottom represents the bottom of the curve; Top represents the top of the curve;

HillSlope represents the absolute value of the maximum slope of the curve.

Experiment 2: Determination of anti-HBV activity in HepG2.2.15 cells

Main reagents:

QIAamp 96 DNA Blood Kit (12) (Qiagen, Cat. No. 51162);

FastStart Universal Probe Master (Roche, article number 04914058001);

Cell–titer Glo detection reagent (Promega, Cat. No. G7573).

Compound dilution: in vitro anti-HBV activity assay and cytotoxicity assay All compounds were serially diluted 3 times, 8 concentrations. The final starting concentration of the test compound was 30 μM, the final starting concentration of the reference compound GLS4 was 1 μM, and the final concentration of DMSO was 0.5%.

HepG2.2.15 cells (4 x 10 ⁴ cells/well) were inoculated into 96-well plates and incubated overnight at 37 ° C, 5% CO ₂ . The next day, fresh culture medium containing different concentrations of compounds was added to the culture wells. On the fifth day, the old culture solution in the culture well was aspirated and fresh culture medium containing different concentrations of the compound was added.

On the eighth day, the supernatant in the culture well was collected for extracting HBV DNA from the supernatant, and qPCR was used to detect the HBV DNA content in the supernatant of HepG2.2.15. After collecting the supernatant, the culture medium and the Cell-titer Glo reagent were added to the culture well, and the chemiluminescence value of each well was detected by a microplate reader.

The activity calculation formula is as follows:

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

Where X represents the logarithm of the concentration, Y represents the effect value, and Y is fitted from the bottom to the top with an S-shape;

Bottom represents the bottom of the curve; Top represents the top of the curve;

HillSlope represents the absolute value of the maximum slope of the curve.

Experiment 3: Determination of cytotoxicity

The cytotoxicity of the test compounds was tested using HepG2 cells, and these cells were incubated for 4 days in the presence of the test compound. Cell rejuvenation was assessed using the resazurin assay.

In the table:

A1 indicates IC50 (μM) at <1;

A2 indicates that the IC50 (μM) is between 1 and 100;

A3 indicates that the IC50 (μM) is >100;

B1 indicates that EC50 (μM) is between <1;

B2 indicates that the EC50 (μM) is between 1 and 100;

B3 indicates that the EC50 (μM) is >100;

Among them, the control compound is:

(See WO2014184350A1)

The results showed that the compound of the present invention was excellent in anti-HBV nucleocapsid assembly activity and anti-HBV activity in vitro, and had lower cytotoxicity.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims (10)

1. A compound of the formula A, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof,

   

   among them,

   R ₁ and R ₂ are each independently selected from the group consisting of hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted having 1- 3 to 3 membered heterocycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, halogen or substituted or unsubstituted having 3 hetero atoms selected from the group consisting of N, S and O have 1-3 options a 5-10 membered heteroaryl group of the following heteroatoms of N, S and O, wherein in R _{1 ,} R ₂ , the substitution is replaced by one or more substituents selected from the group consisting of: -OH , halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, =0, -O-;

   X is CR ₁₁ R ₁₂ or -CR ₁₁ =CR ₁₂ -; wherein R ₁₁ and R ₁₂ are each independently selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or not Substituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted, having 1-3 selected from a 3-10 membered heterocycloalkyl group of a hetero atom of N, S and O, a substituted or unsubstituted C ₆ -C ₁₀ aryl group, substituted or unsubstituted, having 1-3 selected from the group consisting of N, S and a 5-10 membered heteroaryl group of a hetero atom of O, a substituted or unsubstituted C _1-3 alkyl-R ₇ , or a -C(=O)OC _1-4 alkyl group; wherein said R _{7 is} selected from the group consisting of Group: halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted 5-10 membered heteroaryl having 1-2 heteroatoms selected from the group consisting of N, S and O, having 1-3 options a 3-7 membered heterocycloalkyl group of the following heteroatoms of N, S and O, -NR ₉ R ₁₀ , wherein said R ₉ and R ₁₀ are each independently selected from the group consisting of: H, C ₁ -C ₃ alkane Base, halogenated C ₁ -C ₃ alkyl;

   or,

   The R ₁₁ and R ₁₂ together with the adjacent C atoms constitute a substituted or unsubstituted 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, wherein The substitution of a 3-7 membered heterocycloalkyl group is substituted by one or more substituents selected from the group consisting of -OH, halogen, methoxy, -O-, -C(=O)OC _1-4 Alkyl, benzyl, C _1-4 alkyl, halogenated C _1-4 alkyl,

   And, in R ₁₁ and R ₁₂ , the substitution means being substituted with one or more substituents selected from the group consisting of -OH, halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, -OH substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -C(=O)OC _1-4 alkyl;

   Y is a substituted or unsubstituted C ₁ -C ₇ alkylene group or a C ₂ -C ₇ alkenylene group, and in the Y, the substitution means being substituted with one or more substituents selected from the group consisting of C: ₁ -C ₄ alkyl, halogen, -OH, preferably a C ₁ -C ₄ alkyl or -OH;

   Z is selected from the group consisting of NH, O or a bond;

   Ring C is a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 heteroatoms selected from the group consisting of N, S and O, wherein said ring C is selected from the group consisting of Substituted by one or more substituents: C ₁ -C ₃ alkyl, C ₃ -C ₄ cycloalkyl, -CN or halogen;

   Ring B is a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-10 membered heteroaryl group having 1-3 hetero atoms selected from the group consisting of N, S and O; In B, the substitution refers to being substituted with one or more substituents selected from the group consisting of C ₁ -C ₃ alkyl, C ₃ -C ₄ cycloalkyl, -CN or halogen;

   R _a , R _b , R _c , R _d are substituents at any position on Ring B, each independently selected from the group consisting of H, halogen, —CN, hydroxy, amino, carboxy, —(C=O)- Substituted or unsubstituted C ₁ -C ₈ alkyl, substituted or unsubstituted C ₁ -C ₈ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkyne a substituted, unsubstituted or unsubstituted C ₁ -C ₈ alkylamino group, a substituted or unsubstituted C ₁ -C ₈ alkoxy group, a substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl group, substituted or unsubstituted 3 to 3 membered heterocycloalkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, or 1-3 substituted or unsubstituted, selected from the group consisting of the following heteroatoms of N, S and O a 5-10 membered heteroaryl group selected from the group consisting of a hetero atom of N, S and O; in said R _a , R _b , R _c , R _d , said "substituted" means a group selected from the group consisting of Or substituted with a plurality of substituents: halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogenated C ₁ -C ₆ alkoxy, C ₃ -C ₈ cycloalkyl group, a halo-C ₃ -C ₈ cycloalkyl, oxo, -CN, hydroxy, amino, carboxy, C ₆ -C ₁₀ aryl group, a halogenated C ₆ -C ₁₀ aryl Base 5-10 membered heteroaryl groups having 1-3 hetero atoms selected from the group consisting of N, S and O, halogenated 5-10 having 1-3 hetero atoms selected from the group consisting of N, S and O Meta-heteroaryl.
2. The compound according to claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein R ₁ and R ₂ are each independently selected. From the following group: hydrogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted, having 1-3 selected from the group consisting of N, S And a 3-10 membered heterocycloalkyl group of a hetero atom of O, a substituted or unsubstituted C ₆ -C ₁₀ aryl group, or a substituted or unsubstituted 1-3 group selected from the group consisting of N, S and O a 5-10 membered heteroaryl group of an atom, wherein in R _{1 ,} R ₂ , the substitution is substituted with one or more substituents selected from the group consisting of -OH, halogen, C ₁ -C ₆ alkyl , halogenated C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -O-;

   X is CR ₁₁ R ₁₂ or -CR ₁₁ =CR ₁₂ -; wherein R ₁₁ and R ₁₂ are each independently selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or not Substituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted, having 1-3 selected from a 3-10 membered heterocycloalkyl group of a hetero atom of N, S and O, a substituted or unsubstituted C ₆ -C ₁₀ aryl group, substituted or unsubstituted, having 1-3 selected from the group consisting of N, S and a 5-10 membered heteroaryl group of a hetero atom of O, a C _1-3 alkyl-R ₇ , or a -C(=O)OC _1-4 alkyl group; wherein said R _{7 is} selected from the group consisting of halogen, C _1- C ₃ alkyl, substituted or unsubstituted 5-10 membered heteroaryl having 1-2 heteroatoms selected from the group consisting of N, S and O, having 1-3 selected from the group consisting of N, a 3-7 membered heterocycloalkyl of a hetero atom of S and O, -NR ₉ R ₁₀ , wherein said R ₉ and R ₁₀ are each independently selected from the group consisting of: H, C ₁ -C ₃ alkyl, halogenated C ₁ -C ₃ alkyl;

   or,

   The R ₁₁ and R ₁₂ together with the adjacent C atoms constitute a substituted or unsubstituted 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, wherein The substitution of a 3-7 membered heterocycloalkyl group is substituted by one or more substituents selected from the group consisting of -OH, halogen, methoxy, -O-, -C(=O)OC _1-4 Alkyl, benzyl, C _1-4 alkyl, halogenated C _1-4 alkyl,

   And, in R ₁₁ and R ₁₂ , the substitution means being substituted with one or more substituents selected from the group consisting of -OH, halogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, -OH substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, -C(=O)OC _1-4 alkyl;

   R _a , R _b , R _c and R _d are substituents at any position on the benzene ring, as defined in claim 1.
3. The compound according to claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein R ₁ is H, unsubstituted C ₁ - C ₁₀ alkyl, C ₃ -C ₁₀ cycloalkyl, or C ₁ -C ₁₀ alkyl substituted by -OH, =O, -O- or halogen.
4. The compound according to claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein ring C is substituted or unsubstituted 5 A meta or 6-membered heteroaryl group, said substitution being substituted by one or more substituents selected from the group consisting of methyl, -CN or halogen.
5. The compound according to claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein said R _a , R _b , R _c and R _d are each independently selected from the group consisting of H, halogen, -CHF ₂ , -CF ₂ -methyl, -CH ₂ F, -CF ₃ , -OCF ₃ , -CN, -C ₃ -C ₄ ring Alkyl, or -C ₁ -C ₄ alkyl.
6. The compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound of formula A is selected from the group consisting of :

   

   
7. A process for the preparation of a compound according to claim 1 or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound of formula A is of formula VII-1 The compound shown, the method comprising the step (I):

   

   or,

   The compound of formula A shown is a compound of formula VIII-1, said method comprising step (II):

   

   In the step (I) or (II), R _{2 ,} R _a , R _b , R _c , R _d , ring C, and ring B are as defined in claim 1, and m and n are each 1-5. Positive integer

   Wherein R _{3 is} selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₁₀ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkyne a substituted, unsubstituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted 3-10 membered heterocycloalkyl having 1 to 3 heteroatoms selected from the group consisting of N, S and O, substituted or Unsubstituted C ₆ -C ₁₀ aryl, or substituted or unsubstituted 5-10 membered heteroaryl, _1-3 alkyl group having 1-3 heteroatoms selected from the group consisting of N, S and O - R ₇ , —C(=O)OC _1-4 alkyl; wherein said R _{7 is} selected from the group consisting of halogen, C ₁ -C ₃ alkyl, substituted or unsubstituted, and 1-2 selected from the group consisting of a 5-10 membered heteroaryl group of a hetero atom of N, S and O, a 3-7 membered heterocycloalkyl group having 1 to 3 hetero atoms selected from the group consisting of N, S and O, -NR ₉ R ₁₀ Wherein R ₉ and R ₁₀ are each independently selected from the group consisting of: H, C ₁ -C ₃ alkyl, halogenated C ₁ -C ₃ alkyl.
8. A pharmaceutical composition comprising (1) the compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof; (2) A pharmaceutically acceptable carrier.
9. The compound shown below:

   
10. The use of a compound according to claim 1 or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 8 For the preparation of a medicament for preventing and/or treating hepatitis B virus infection.