CN118666868A - Intracyclic thioguanamide-arylamide compounds and use thereof in treating hepatitis B - Google Patents

Abstract

An application of an inner ring thioether amide-aryl amide compound, a pharmaceutical composition containing the compound and a compound or pharmaceutical composition of the compound in treating hepatitis B. Specifically, it is a compound having a structure represented by the formula (L) or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, which is useful as an HBV replication inhibitor.

Description

Intracyclic thioguanamide-arylamide compounds and use thereof in treating hepatitis B

Technical Field

The present invention belongs to the field of medicine, and in particular, relates to an endocyclic thioguanamide-arylamide compound and its use as a medicine for treating hepatitis B.

Background Art

Hepatitis B virus (HBV) is an enveloped, partially double-stranded DNA (dsDNA) virus of the hepadnaviridae family. Its genome contains 4 overlapping reading frames: prenuclear/nuclear genes, polymerase genes, UM and S genes (which encode three envelope proteins), and X genes. Before infection, the partial double-stranded DNA genome is converted into covalently closed circular DNA (cccDNA) in the host cell nucleus (opened DNA, rcDNA) and the viral mRNA is transcribed. Once encapsidated, the pregenomic RNA (pgRNA) (which also encodes core protein and Pol) is used as a template for reverse transcription, which regenerates the partial dsDNA genome (rcDNA) in the nucleocapsid.

HBV has caused epidemics in parts of Asia and Africa, and it is endemic in China. HBV has infected approximately 2 billion people worldwide, of whom approximately 350 million have developed chronic infection. The virus causes hepatitis B disease and chronic infection is associated with a high increased risk of developing cirrhosis and liver cancer.

Hepatitis B virus is transmitted through exposure to infectious blood or body fluids, and viral DNA has been detected in the saliva, tears, and urine of chronic carriers with high titers of DNA in the serum.

Although an effective and well-tolerated vaccine exists, immediate treatment options are currently limited to interferon and the following antiviral drugs: tenofovir, lamivudine, adefovir, entecavir, and telbivudine.

In addition, heteroaryldihydropyrimidines (HAPs) were identified as a class of HBV inhibitors in tissue culture and animal models (Weber et al., Antiviral Res. 54:69-78).

WO 2013/006394 (published on January 10, 2013) and WO 2013/096744 (published on June 27, 2013) also disclose sulfamoyl-aryl amides involved in anti-HBV activity.

However, problems encountered among these direct HBV antiviral drugs include toxicity, mutagenicity, lack of selectivity, poor efficacy, poor bioavailability, and difficulty in synthesis.

Therefore, in order to overcome the above defects, it is necessary to develop HBV inhibitors with advantages such as high potency and lower toxicity.

Summary of the invention

The object of the present invention is to provide a class of compounds with novel structures which can be used as HBV inhibitors.

In a first aspect, the present invention provides a compound as shown in formula L, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof,

Wherein, n is 1, 2, 3, 4 or 5;

is a substituted or unsubstituted five-membered or six-membered ring, wherein the five-membered or six-membered ring optionally contains one or more heteroatoms selected from the group consisting of O, S, N or P; the substitution refers to the replacement of hydrogen atoms on the group by one or more substituents selected from the group consisting of C1-C3 alkyl (especially methyl), C3-C4 cycloalkyl, cyano, or halogen;

is a substituted or unsubstituted five-membered or six-membered aromatic ring, or a substituted or unsubstituted five-membered or six-membered aromatic heterocycle;

T is NR ₆ or O;

X is selected from the group consisting ^{of -CRaRb- , -CRaRb} ( ^{CRcRd )} -, or ^-CRaRb ( ^NRc ) ^- ;

Where ^Ra is R ^b is selected from the group consisting of H, halogen, substituted or unsubstituted C ₁ -C ₈ alkyl;

R ^c and R ^d are each independently H, halogen, -CN, hydroxyl, amino, carboxyl, -(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 alkynyl, substituted or unsubstituted C ₁ -C ₈ alkylamino, substituted or unsubstituted C ₁ -C ₈ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl;

or ^Ra and ^Rb and the carbon atoms to which they are connected together form a substituted C3-C8 cycloalkyl, or a substituted 4-8 membered heterocycle (the heterocycle includes 1-3 heteroatoms selected from N, S or O);

or ^Ra and ^Rc and the carbon atoms to which they are connected together form a substituted C3-C8 cycloalkyl, or a substituted 4-8 membered heterocycle (the heterocycle includes 1-3 heteroatoms selected from N, S or O);

or ^Ra and ^R5 and the carbon atom and nitrogen atom to which they are connected together form a substituted 4-8 membered heterocyclic ring (the heterocyclic ring includes 1-3 heteroatoms selected from N, S or O);

Wherein, in addition to conventional substituents, the formed cycloalkyl or heterocyclic group also includes at least one of the following substituents:

W is selected from the group consisting of NH, O, S, CH ₂ ;

Y is a substituted or unsubstituted C1-C7 alkylene group, or a substituted or unsubstituted C2-C7 alkenylene group, wherein the substituent is selected from the following group: a C1-C4 alkyl group, a hydroxyl group;

Z is selected from the following group: O, S, N or P, or Z is a C-C single bond (i.e., Z is nothing); in another preferred embodiment, when T is O, Z is a C-C single bond;

R ¹ , R ² , R ³ and R ⁴ are each independently selected from the following group: H, halogen, cyano, substituted or unsubstituted C3-C4 cycloalkyl, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy; wherein the substitution refers to the substitution of hydrogen atoms on the group by one or more substituents selected from the following group: halogen, C1-C4 alkyl (such as difluoromethyl, difluoroethyl, monofluoromethyl, trifluoromethyl, trifluoromethoxy);

R ⁵ and R ⁶ are each independently selected from the following group: H, halogen, -CN, hydroxyl, amino, carboxyl, -(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 alkynyl, substituted or unsubstituted C ₁ -C ₈ alkylamino, substituted or unsubstituted C ₁ -C ₈ alkoxy, substituted or unsubstituted C ₃ -C ₈ cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl having 1-3 heteroatoms selected from the following group of N, S and O;

R ₁₀ is selected from the group consisting of: -NHCHR ₇ R ₈ ;

R ⁷ and R ⁸ are each independently selected from the following group: H, halogen, -CN, hydroxyl, amino, carboxyl, -(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 alkynyl, substituted or unsubstituted C ₁ -C ₈ alkylamino, substituted or unsubstituted C ₁ -C ₈ alkoxy, substituted or unsubstituted C ₃ -C 8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclic group having 1-3 heteroatoms selected from the following group: N, S and O; or R ⁷ and R ⁸ and the carbon atoms to which they are connected together form a substituted C 3 -C ₈ cycloalkyl, or a substituted 4-8 membered heterocyclic ring (the heterocyclic ring includes 1-3 heteroatoms selected from N, S or O);

V is selected from the following group: a chemical bond, NH, -( ^{CReRf )} _m- , -NH ^{( CReRf} ) _m- ; wherein each ^Re and ^Rf is independently selected from the following group: H, halogen, -CN, hydroxyl, amino, carboxyl, substituted or unsubstituted _C1 - _C8 alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C1 - _C8 alkylamino, substituted or unsubstituted _C1 - _C8 alkoxy, substituted or unsubstituted _C3 - _C8 cycloalkyl; or ^Re and ^Rf and the carbon atoms to which they are connected together form a substituted or unsubstituted C3-C8 cycloalkyl, or a substituted or unsubstituted 4-8 membered heterocycle (the heterocycle includes 1-3 heteroatoms selected from N, S or O);

Unless otherwise specified, the "substituted" refers to replacement by one or more (e.g., 2, 3, 4, etc.) substituents selected from the following group: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxyl, amino, carboxyl, a group selected from the following group which is unsubstituted or substituted by one or more substituents selected from the following group: C6-C10 aryl, halogenated C6-C10 aryl, a 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, and a halogenated 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O; the substituents are selected from the following group: halogen, C1-C6 alkoxy.

In another preferred embodiment, the X is a group selected from the following group:

wherein p and p' are each independently selected from the following groups: 0, 1, 2, 3 or 4; and the is a 3-8 membered ring; W is selected from N or CH;

R ₉ is In another preferred embodiment, the compound of formula I has a structure as shown in formula II-A or formula II-B:

wherein X ₁ is -CR ₁₁ = or -N =, X ₂ is -NR ₁₁ -; and R ₁₁ is H or a C ₁ -C ₄ alkyl group.

In another preferred embodiment, the X ₂ is -NCH ₃ -.

In another preferred embodiment, the compound of formula I has a structure as shown in formula III-A or III-B:

In another preferred embodiment, the compound of formula I has a structure shown in the following formula IV:

In another preferred embodiment, the compound of formula I has a structure shown in the following formula IV:

In another preferred embodiment, p and p' are each independently selected from the following group: 1 or 2; W is each independently selected from the following group: N or CH;

R ₉ is Wherein, V is selected from the following group: a chemical bond, NH, -( ^{CReRf )} _m- , -NH ^{( CReRf} ) _m- ; wherein, each ^Re and ^Rf is independently selected from the following group: H, halogen, substituted or unsubstituted _C1 - _C8 alkyl, substituted or unsubstituted _C3 - _C8 cycloalkyl; or ^{Re and Rf and} the carbon atoms to which they are connected together form a substituted or unsubstituted C3 _{- C8} cycloalkyl, or a substituted or unsubstituted 4-8 membered heterocycle (the heterocycle includes 1-3 heteroatoms selected from N, S or O).

In another preferred embodiment, the compound is the compound described in Table 1.

The second aspect of the present invention provides a method for preparing the compound according to the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, the method comprising the steps of:

In an inert solvent, a compound of formula L1 is reacted with L2 to obtain a compound of formula I; wherein R is a leaving group, and the definitions of the remaining groups are as described in the first aspect of the present invention.

In another preferred embodiment, the compound of formula I is a compound of formula VII-1, and the method comprises the steps of:

Wherein, Rg is selected from the following group: H, halogen, -CN, hydroxyl, amino, carboxyl, -CC=O)-substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C8 alkylamino, substituted or unsubstituted C1-C8 alkoxy, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted 3-10 membered heterocycloalkyl having 1-3 heteroatoms selected from the following group N, S and O, substituted or unsubstituted C6-C10 aryl, or substituted or unsubstituted 5-10 membered heteroaryl having 1-3 heteroatoms selected from the following group N, S and O;

The definitions of the various groups are as described above.

In another preferred embodiment, the compound of formula I is a compound of formula II-2, and the method comprises the steps of:

In another preferred embodiment, the compound of formula I is a compound of formula III-7, and the method comprises the steps of:

wherein Rg is selected from the group consisting of H, halogen, -CN, hydroxyl, amino, carboxyl, -(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 alkynyl, substituted or unsubstituted C _{1 -C 8 alkylamino, substituted or unsubstituted C 1} -C ₈ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted 3-10 membered heterocycloalkyl having 1 to 3 heteroatoms selected from N, S and O, substituted or unsubstituted C ₆ -C ₁₀ aryl, or substituted or unsubstituted 5-10 membered heteroaryl having 1 to 3 heteroatoms selected from the group consisting of N, S and O;

The definitions of the various groups are as described above.

In another preferred embodiment, the compound of formula I is a compound of formula IV-5 and IV-6, and the method comprises the steps of:

In another preferred embodiment, the compound of formula I is a compound of formula V-4, and the method comprises the steps of:

In another preferred embodiment, the compound of formula I is a compound of formula VI-5 and VI-6, and the method comprises the steps of:

In another preferred embodiment, the compound of formula I is a compound of formula VII-4, and the method comprises the steps of:

In another preferred embodiment, the compound of formula I is a compound of formula VIII-4 and VIII-5, and the method comprises the steps of:

The third aspect of the present invention provides a pharmaceutical composition comprising (1) a compound as described in the first aspect of the present invention or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof; and (2) a pharmaceutically acceptable carrier.

The fourth aspect of the present invention provides a use of the compound as described in the first aspect of the present invention or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition as described in the third aspect of the present invention, for preparing a medicament for preventing and/or treating hepatitis B virus infection.

The fifth aspect of the present invention provides a hepatitis B virus inhibitor, characterized in that the inhibitor comprises the compound described in the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate.

The sixth aspect of the present invention provides a method for preventing and/or treating hepatitis B, comprising the steps of administering to a patient in need thereof the compound described in the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition described in the fourth aspect of the present invention.

The seventh aspect of the present invention provides a method for inhibiting the replication of hepatitis B virus, characterized in that it comprises the steps of: contacting the compound described in the first aspect of the present invention, or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate with hepatitis B virus, thereby inhibiting the replication of hepatitis B virus.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

DETAILED DESCRIPTION

After extensive and in-depth research, the inventors have discovered a new type of compound with excellent therapeutic effect on hepatitis B. On this basis, the inventors have completed the present invention.

definition

As used herein, the term "alkyl" includes linear or branched alkyl groups. For example, _C1 - _C8 alkyl groups represent linear or branched alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.

As used herein, the term "alkenyl" includes straight or branched alkenyl groups. For example, _C2 - _C6 alkenyl refers to straight or branched alkenyl groups having 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or similar groups.

As used herein, the term "alkynyl" includes straight or branched chain alkynyl groups. For example, _C2 - _C6 alkynyl refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.

As used herein, the term "C ₃ -C ₁₀ cycloalkyl" refers to a cycloalkyl group having 3 to 10 carbon atoms. It may be a monocyclic ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may also be a bicyclic ring, such as a bridged ring or a 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, propylamino, isopropylamino, butylamino, isobutylamino, tert-butylamino, dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, di-tert-butylamino and the like.

As used herein, the term "C ₁ -C ₈ alkoxy" refers to a straight or branched alkoxy group having 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, isopropoxy, 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" refers to a saturated or partially saturated cyclic group having 3-10 atoms, wherein 1-3 atoms are heteroatoms selected from the group consisting of N, S and O. It may be a monocyclic or bicyclic form, such as a bridged ring or a spirocyclic form. Specific examples may include oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl and pyrrolidinyl, etc.

As used herein, the term "C ₆ -C ₁₀ aryl group" refers to an aryl group having 6 to 10 carbon atoms, for example, a phenyl group, a 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 a cyclic aromatic group having 5-10 atoms, of which 1-3 atoms are heteroatoms selected from the group consisting of N, S and O. It may be a monocyclic ring or a condensed ring. Specific examples may include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2,4)-triazolyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl and the like.

Unless otherwise specified as "substituted or unsubstituted", the groups described in the present invention may be substituted by substituents selected from the following groups: halogen, nitrile, nitro, hydroxyl, amino, 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, halogenated C ₂ -C ₆ alkenyl, halogenated C ₂ -C ₆ alkynyl, halogenated 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-carbonyl, C ₃ -C ₆ cycloalkyl-carbonyl, C ₁ -C 6 _6- alkyl-sulfonyl, etc.

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

Unless otherwise specified, the structural formulas described in the present invention are intended to include all isomeric forms (such as enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers or geometric isomers (or conformational isomers) are all within the scope of the present invention.

As used herein, the term "tautomer" means that structural isomers with different energies can interconvert across a low energy barrier. For example, proton tautomers (i.e., prototropic) include interconversion via proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion via reorganization of some bonding electrons.

As used herein, the term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

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

Active ingredients

As used herein, "compounds of the present invention" refers to compounds represented by formula L, and also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula L:

As used herein, "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, 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, picric acid, methanesulfonic acid, benzenesulfonic acid, benzenesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid.

In another preferred embodiment, the A ring, B ring, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , and R ₆ are each independently a group corresponding to each compound in Table 1.

Preferred compounds of the present invention are shown in Table 1.

Pharmaceutical compositions and methods of administration

Since the compounds of the present invention have excellent inhibitory activity against hepatitis B virus (HBV), the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to prevent and/or treat (stabilize, alleviate or cure) hepatitis B virus infection or to prevent and/or treat (stabilize, alleviate or cure) hepatitis B virus-related diseases (e.g., hepatitis B, progressive liver fibrosis, inflammation and necrosis leading to cirrhosis, end-stage liver disease, and ethyl liver cancer).

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration or subcutaneous administration).

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 with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into 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 tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

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

When administered in combination, the pharmaceutical composition also includes 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 compounds (e.g., anti-HBV agents) can be used simultaneously, separately, or sequentially with the compounds of the present invention to prevent and/or treat HBV infection or HBV-related diseases.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

The main advantages of the present invention include:

(1) The compounds of the present invention have novel structures and excellent anti-HBV infection effects. In the present application, the existing endocyclic sulfoxide amide-aryl amide compounds are transformed into endocyclic sulfoguanamide-aryl amide compounds, so that they can better interfere with the capsid protein assembly process, thereby inhibiting the activity or expression of HBV.

(2) The compounds of the present invention have very low toxicity to normal cells and can therefore be used in therapeutic subjects within a wide dosage range.

(3) The compounds of the present invention have good drugability. Compared with existing compounds, the compounds of the present invention have better solubility and show good bioavailability in in vivo experiments. The bioavailability of some compounds reaches or exceeds 70%. In addition, compared with existing compounds, the compounds of the present invention are very easy to prepare pharmaceutically acceptable salts, which is helpful for further forming preparations.

(4) The compounds of the present invention and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to prevent and/or treat hepatitis B virus-related diseases (e.g., hepatitis B, progressive liver fibrosis, inflammation and necrosis leading to cirrhosis, end-stage liver disease, and hepatocellular carcinoma).

The present invention will be further described below in conjunction with specific implementation. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually carried out under conventional conditions or under conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight. The raw materials or instruments used in the embodiments of the present invention are commercially available unless otherwise specified.

Biological Example--Anti-HBV Activity Experiment

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

Main reagents and raw materials:

The C150 protein was expressed and purified by WuXi AppTec;

FL was purchased from Thermo Fisher Scientific.

Protein fluorescent labeling:

Add 150 μL of 2% w/v skim milk to each well of a 96-well plate and incubate at room temperature for 2 hours. Remove the skim milk, rinse with deionized water, dry, and store at room temperature. Desalt the C150 protein (3 mg per tube) using a 5 ml Hitrap desalting column. Add 50 mM 20 μl of FL fluorescent dye was mixed evenly and incubated at 4°C in the dark overnight. The fluorescent dye that was not bound to C150 was removed by Sephadex G-25 gel filtration. The fluorescent labeling efficiency of C150 was calculated as follows:

in,

[ FL] indicates the concentration of fluorescent marker;

[C150Bo] indicates the concentration of fluorescent labeled protein;

A504 represents the absorbance value at a wavelength of 504 nM;

A280 represents the absorbance value at a wavelength of 280 nM;

M ^-1 represents the reciprocal of molar concentration.

Compound Dilution:

Compound stocks were diluted to 6 mM with DMSO, then to 600 [mu]M with 50 mM HEPES, and then further serially diluted 3-fold to 8 concentrations with 10% DMSO/50 mM HEPES.

Dilute C150Bo to 2μM with 50mM HEPES. Add 37.5μL C150Bo and 2.5μL of each concentration of compound to a 96-well reaction plate, mix well, and incubate at room temperature for 15 minutes. Add 10μl of 750mM NaCl/50mM HEPES to the reaction wells, and the final concentration of NaCl is 150mM.

For the 0% protein assembly control well, 10 μL of 50 mM HEPES was added, and the final concentration of NaCl was 0 mM.

For the 100% protein assembly control well, add 10 μL of 5M NaCl/50 mM HEPES, and the final concentration of NaCl is 1M.

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

Data analysis

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

The _IC50 value was calculated by Prism software using the following equation:

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

in,

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

Bottom indicates the bottom of the curve;

Top means Top means the top of the curve;

HillSlope means: 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, Cat. No. 04914058001);

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

Compound dilution: All compounds in the in vitro anti-HBV activity test and cytotoxicity test were serially diluted 3-fold to 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 (4×10 ⁴ cells/well) were seeded into 96-well plates and cultured overnight at 37°C, 5% CO _2. On the second day, fresh culture medium containing different concentrations of compounds was added to the culture wells. On the fifth day, the old culture medium in the culture wells was removed and fresh culture medium containing different concentrations of compounds was added.

On the eighth day, the supernatant in the culture wells was collected for extraction of HBV DNA in the supernatant, and the HBV DNA content in the supernatant of HepG2.2.15 was detected by qPCR. After the supernatant was collected, the culture medium and Cell-titer Glo reagent were added to the culture wells, and the chemiluminescence value of each well was detected by an enzyme-labeled instrument.

The activity calculation formula is as follows:

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

in,

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

Bottom represents the bottom of the curve;

Top indicates the top of the curve;

HillSlope means: the absolute value of the maximum slope of the curve.

Experiment 3: Cytotoxicity assay

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

The results show that the compounds of the present invention have good in vitro anti-HBV nucleocapsid assembly activity and anti-HBV activity, and low cytotoxicity.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (13)

1. A compound represented by formula L, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof,

Wherein n is 1,2, 3,4 or 5;

Is a substituted or unsubstituted five or six membered ring, wherein the five or six membered ring optionally contains one or more heteroatoms selected from the group consisting of: o, S, N or P; the substitution means that the hydrogen atom on the group is substituted with one or more substituents selected from the group consisting of: C1-C3 alkyl (especially methyl), C3-C4 cycloalkyl, cyano, or halogen;

is a substituted or unsubstituted five-membered or six-membered aromatic ring, or a substituted or unsubstituted five-membered or six-membered aromatic heterocyclic ring;

T is NR ₆ or O;

X is selected from the group consisting of: -CR ^aR^b-、-CR^aR^b(CR^cR^d) -, or-CR ^aR^b(NR^c) -;

wherein R ^a is R ^b is selected from the group consisting of: H. halogen, substituted or unsubstituted C ₁-C₈ alkyl;

R ^c and R ^d are each independently H, halogen, -CN, hydroxy, amino, carboxy, - (c=o) -substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₂-C₆ alkenyl, substituted or unsubstituted C ₂-C₆ alkynyl, substituted or unsubstituted C ₁-C₈ alkylamino, substituted or unsubstituted C ₁-C₈ alkoxy, substituted or unsubstituted C ₃-C₁₀ cycloalkyl;

Or R ^a and R ^b together with the carbon atom to which they are attached form a substituted C3-C8 cycloalkyl group, or a substituted 4-8 membered heterocyclic ring (said heterocyclic ring comprising 1-3 heteroatoms selected from N, S or O);

Or R ^a and R ^c together with the carbon atom to which they are attached form a substituted C3-C8 cycloalkyl group, or a substituted 4-8 membered heterocyclic ring (said heterocyclic ring comprising 1-3 heteroatoms selected from N, S or O);

Or R ^a and R ⁵ together with the carbon and nitrogen atoms to which they are attached form a substituted 4-8 membered heterocyclic ring (said heterocyclic ring comprising 1-3 heteroatoms selected from N, S or O);

wherein, in addition to conventional substituents, the cycloalkyl or heterocyclyl groups formed include at least one of the following substituents:

W is selected from the group consisting of: NH, O, S, CH ₂;

y is a substituted or unsubstituted C1-C7 alkylene group, or a substituted or unsubstituted C2-C7 alkenylene group, wherein the substituents are selected from the group consisting of: C1-C4 alkyl, hydroxy;

Z is selected from the group consisting of: o, S, N or P, or Z is a C-C single bond (i.e., Z is absent);

R ¹,R²,R³ and R ⁴ are each independently selected from the group consisting of: H. halogen, cyano, substituted or unsubstituted C3-C4 cycloalkyl, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C1-C4 alkoxy; wherein the substitution means that a hydrogen atom on the group is substituted with one or more substituents selected from the group consisting of: halogen, C1-C4 alkyl (e.g., difluoromethyl, difluoroethyl, monofluoromethyl, trifluoromethyl, trifluoromethoxy);

R ⁵、R⁶ are each independently selected from the group consisting of: H. halogen, -CN, hydroxy, amino, carboxy, - (c=o) -substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₂-C₆ alkenyl, substituted or unsubstituted C ₂-C₆ alkynyl, substituted or unsubstituted C ₁-C₈ alkylamino, substituted or unsubstituted C ₁-C₈ alkoxy, substituted or unsubstituted C ₃-C₈ cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S and O;

R ₁₀ is selected from the group consisting of: -NHCHR ₇R₈;

R ⁷、R⁸ are each independently selected from the group consisting of: H. halogen, -CN, hydroxy, amino, carboxy, - (c=o) -substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₂-C₆ alkenyl, substituted or unsubstituted C ₂-C₆ alkynyl, substituted or unsubstituted C ₁-C₈ alkylamino, substituted or unsubstituted C ₁-C₈ alkoxy, substituted or unsubstituted C ₃-C₈ cycloalkyl, substituted or unsubstituted 3-8 membered heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S and O; or R ⁷ and R ⁸ together with the carbon atom to which they are attached form a substituted C3-C8 cycloalkyl group, or a substituted 4-8 membered heterocyclic ring (said heterocyclic ring comprising 1-3 heteroatoms selected from N, S or O);

V is selected from the group consisting of: a bond, NH, - (CR ^eR^f)_m-、-NH(CR^eR^f)_m -; wherein each R ^e and R ^f is independently selected from the group consisting of H, halogen, -CN, hydroxy, amino, carboxy, substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₂-C₆ alkenyl, substituted or unsubstituted C ₂-C₆ alkynyl, substituted or unsubstituted C ₁-C₈ alkylamino, substituted or unsubstituted C ₁-C₈ alkoxy, substituted or unsubstituted C ₃-C₈ cycloalkyl, or R ^e and R ^f together with the carbon atoms to which they are attached form a substituted or unsubstituted C3-C8 cycloalkyl, or a substituted or unsubstituted 4-8 membered heterocycle (said heterocycle comprising 1 to 3 heteroatoms selected from N, S or O), m is selected from the group consisting of 1, 2, 3, 4 or 5;

R ₇,R₈ is not defined

Unless otherwise specified, "substituted" means substituted with one or more (e.g., 2, 3, 4, etc.) substituents selected from the group consisting of: halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C8 cycloalkyl, halogenated C3-C8 cycloalkyl, oxo, -CN, hydroxy, amino, carboxy, a group selected from the group consisting of: C6-C10 aryl, halogenated C6-C10 aryl, 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O, halogenated 5-10 membered heteroaryl having 1-3 heteroatoms selected from N, S and O; the substituents are selected from the group consisting of: halogen, C1-C6 alkoxy.

2. The compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein X is a group selected from the group consisting of:

Wherein p and p' are each independently selected from the group consisting of: 0.1, 2,3 or 4; and is described as Is a 3-8 membered ring; w is selected from N or CH;

R ₉ is

3. 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 I has a structure according to formula II-a or formula II-B:

Wherein X ₁ is-CR ₁₁ =or-n=, X ₂ is-NR ₁₁ -; and R ₁₁ is H or C ₁-C₄ alkyl.

4. The compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein X ₂ is-NCH ₃ -.

5. 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 I has the structure of formula III-a or formula III-B as follows:

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 I has the structure of formula IV:

7. 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 I has the structure of formula V:

8. The compound of any one of claims 1-7, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein p and p' are each independently selected from the group consisting of: 1 or 2; each W is independently selected from the group consisting of: n or CH;

R ₉ is Wherein V is selected from the group consisting of: a bond, NH, - (CR ^eR^f)_m-、-NH(CR^eR^f)_m -; wherein each R ^e and R ^f is independently selected from the group consisting of H, halogen, substituted or unsubstituted C ₁-C₈ alkyl, substituted or unsubstituted C ₃-C₈ cycloalkyl, or R ^e and R ^f together with the carbon atoms to which they are attached form a substituted or unsubstituted C ₃-C₈ cycloalkyl, or a substituted or unsubstituted 4-8 membered heterocyclic ring (said heterocyclic ring comprising 1-3 heteroatoms selected from N, S or O).

9. The compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is a compound as set forth in table 1:

10. A process for the preparation of a compound of formula L, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, according to claim 1, wherein the compound of formula L is a compound of formula VII-1, the process comprising the steps of:

In an inert solvent, using a compound of formula L1 with Reacting to obtain a compound of formula L;

Wherein R is a leaving group and the remaining groups are as defined in claim 1.

11. A pharmaceutical composition comprising (1) a compound of claim 1, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate, or solvate thereof; (2) a pharmaceutically acceptable carrier.

12. Use of a compound according to any one of claims 1 to 8, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 10, for the preparation of a medicament for the prophylaxis and/or treatment of a hepatitis b virus infection.

13. A hepatitis b virus inhibitor, comprising a compound according to any one of claims 1 to 8, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof.