Pyrrolo-ring compound as well as preparation method and application thereof

Abstract

The invention relates to a pyrrolo-ring compound as well as a preparation method and application thereof. The structure of the compound is shown as a formula (I). The compound is a brand-new anti-HBV (Hepatitis B Virus) compound. The invention also relates to a pharmaceutical composition containing the compound as an active component, and the pharmaceutical composition can be used for treating and / or preventing HBV infection and has important application value. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a compound for treating HBV infection, a preparation method and a use thereof. Background Art

[0002] Hepatitis B virus (HBV) causes hepatitis B and remains one of the most concerning viruses worldwide. Currently, more than 250 million people worldwide are carriers of the HBV, which increases the risk of major liver diseases including cirrhosis, permanent scarring of the liver, and liver failure and cancer. The World Health Organization estimates that more than 780,000 people die from HBV each year. According to the Hepatitis B Foundation, HBV accounts for 80% of the factors that induce liver cancer, and the five-year survival rate of liver cancer patients is typically only 15%.

[0003] Despite the availability of effective vaccines, vaccination coverage remains unsatisfactory in areas where HBV is endemic. [1] . Currently, there are two main types of drugs approved for the treatment of chronic hepatitis B. One is interferon (IFN-α) and its pegylated form (Peg-IFN-α). IFN-α is an immunomodulator that can induce the expression of interferon-stimulated genes (ISGs) in a non-specific manner. This gene encodes constitutive or secreted proteins with direct or indirect antiviral properties, thereby promoting the differentiation or activation of immune cells. [2] After 48 weeks of subcutaneous IFN-α, the efficacy rate in HBV patients was only 25%. [3]. In addition to the low response rate, IFN-α also has other shortcomings, such as the need for injection, severe flu-like symptoms and other adverse reactions, and contraindications for patients with compensated cirrhosis, severe hepatitis, and combined autoimmune and psychological diseases. Another type of approved drug is nucleoside analogs (NAs). Currently, there are 5 approved NAs, including Lamivudine (LMV), Telbivudine (Ldt), Adefovirdipivoxil (ADV), Tenofovir (TFV), and Entecavir (ETV). NAs directly inhibit the activity of HBV polymerase, resulting in a reduction in viral particles, thereby interrupting the cycle of nucleocapsids to the nucleus of infected cells, and theoretically reducing the expression of cccDNA. However, NAs treatment cannot inhibit the de novo synthesis of cccDNA in newly infected cells, which indicates that residual viral particles during antiviral treatment will lead to infection of new cells and reconstruction of the cccDNA library. Once patients stop taking the drug on their own, virological rebound may occur. Therefore, chronic hepatitis B virus infection rarely achieves functional cure, and most patients need to take drugs for life, but long-term use of NAs drugs can easily cause the virus to mutate and develop drug resistance.

[0004] Based on the inadequacies of current therapies, some drugs targeting the HBV replication cycle or enhancing the human immune response are being developed and have entered the clinical research stage. Among them, the core protein in the HBV replication process is crucial for the packaging and reverse transcription of HBV pgRNA. The molecules developed for this protein are called core protein allosteric modulators or capsid assembly modulators (CpAMs). According to the mechanism of action of CpAMs, they can be divided into two categories: Class I CpAMs represented by heteroaryldihydropyrimidines (HAPs), whose mechanism of action is to enhance the kinetics of nucleocapsid formation, leading to incorrect assembly of nucleocapsids; Class II CpAMs represented by the structural types of phenylacrylamides (PPAs) and sulfonylbenzamides (SBAs), whose mechanism of action is to accelerate nucleocapsid assembly and form nucleocapsids with normal morphology but lacking viral pgRNA and HBV polymerase. [4] .

[0005] The antiviral effects of nucleocapsid inhibitors BAY41-4109 (HAP) and JNJ-632 (SBA) with different mechanisms were studied in primary human hepatocytes. It was found that CpAMs could not only inhibit HBV replication, but also inhibit HBV RNA transcription and antigen production, suggesting that CpAMs have a dual mechanism of action to inhibit the early and late stages of the virus. [5-6]As an early-stage CpAMs molecule, NVR3-778 (SBA) showed strong anti-HBV activity, with an EC50 of 0.4 μM in HepG2.2.15 cells. Similarly, it also showed effective anti-HBV activity in a humanized liver mouse model of HBV infection. [7] With the advancement of science and technology, drug developers have designed and screened out molecular structures with in vitro anti-HBV activity up to nM level, such as GLP-26 and RG7907.

[0006] Currently, most anti-HBV drugs show good in vitro anti-HBV activity, but after entering the clinic, some drugs have failed in clinical trials due to low efficacy and some adverse reactions such as ALT elevation and recurrence after drug withdrawal. Therefore, although there are many drugs and methods for treating HBV, molecules with new structural types and new mechanisms of action for achieving hepatitis B cure are still urgently needed in the field of HBV treatment. Summary of the invention

[0007] The present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof:

[0008]

[0009] wherein X1 is selected from -NH- or -CH2-;

[0010] R1 is selected from -H, deuterium, C 1-8 Alkyl, C 3-8 Cycloalkyl, C 1-8 Haloalkyl, C 1-8 Deuterated alkyl, C 1-8 Alkoxy, C 1-8 Deuterated alkoxy, halogen, -CN, -OH, -NH2;

[0011] R2 is selected from unsubstituted or substituted by one or more R a Substituted C 6-10 Aryl, unsubstituted or substituted with one or more R b Substituted 5-10 membered heteroaryl; the 5-10 membered heteroaryl contains 1-3 heteroatoms selected from N, O and S;

[0012] Each R a , R b are independently selected from the same or different halogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 1-8 Haloalkoxy, -CN, -NH2;

[0013] The compound of formula (I) does not include the following compounds

[0014]

[0015] In some embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, R1 is selected from -H, deuterium, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Deuterated alkyl, C 3-6 Cycloalkyl, halogen, -CN, -OH, -NH2.

[0016] In some embodiments, R1 is selected from C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Deuterated alkyl, halogen, -CN, -NH2.

[0017] In some embodiments, R1 is selected from C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Deuterated alkyl, halogen, -CN, -NH2.

[0018] In some embodiments, R1 is selected from C 1-2 Alkyl, C 3-4 Alkyl, C 1-2 Fluorinated alkyl, halogen.

[0019] In some embodiments, R1 is selected from methyl, ethyl, trifluoromethyl, -Cl, and -Br.

[0020] In some embodiments, R1 is selected from methyl.

[0021] In some embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted 5-6 membered heteroaryl; the 5-6 membered heteroaryl contains 1-3 heteroatoms selected from N, O and S; each R a , R b are independently selected from the same or different halogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 1-8 Haloalkoxy, -CN, -NH2.

[0022] In some embodiments, R2 is selected from unsubstituted or substituted with one or more Ra Substituted phenyl, unsubstituted or replaced by one or more R b Substituted pyridyl or pyrimidinyl; each R a , R b are independently selected from the same or different halogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 1-8 Haloalkoxy, -CN, -NH2.

[0023] In some embodiments, R2 is selected from unsubstituted or substituted with one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted pyridyl or pyrimidinyl; each R a , R b are independently selected from the same or different halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, -CN, -NH2.

[0024] In some embodiments, R2 is selected from unsubstituted or substituted with one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted pyridyl or pyrimidinyl; each R a , R b are independently selected from the same or different halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, -CN, -NH2.

[0025] In some embodiments, R2 is selected from unsubstituted or substituted with one or more R a Substituted phenyl; each R a are independently selected from the same or different halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, -CN, -NH2.

[0026] In some embodiments, R2 is selected from unsubstituted or substituted with one or more R a Substituted phenyl; each R a Each is independently selected from the same or different F, Cl, Br, methyl, ethyl, isopropyl, fluoromethyl, fluoroethyl, fluoromethoxy, fluoroethoxy, -CN, -NH2.

[0027] In some embodiments, R2 is selected from unsubstituted or substituted with 1, 2 or 3 R aSubstituted phenyl; each R a Each is independently selected from the same or different F, Cl, Br, -CN, -NH2.

[0028] Most preferably, R2 is selected from

[0029] As a preferred technical solution of the present invention, the above-mentioned compound, or its pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer, the formula I has the structure shown in formula II:

[0030]

[0031] Wherein, the definitions of R1 and R2 are as shown above.

[0032] In some specific embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, the formula I has the structure shown in formula IIa:

[0033]

[0034] Among them, R1, R a The definition of is as shown above, and m is 1, 2 or 3.

[0035] In some specific embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, the formula I has the structure shown in formula IIa-1:

[0036]

[0037] Among them, R a The definition of is as shown above, and m is 1, 2 or 3.

[0038] As a preferred technical solution of the present invention, the above compound, or its pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer, the formula I has the structure shown in formula III:

[0039]

[0040] The compound of formula (III) does not include the following compounds

[0041]

[0042] Wherein, the definitions of R1 and R2 are as shown above.

[0043] In some specific embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, the formula I has the structure shown in formula IIIa:

[0044]

[0045] The compound of formula (IIIa) does not include the following compounds

[0046]

[0047] Among them, R a , R1 is as defined above, and n is 1, 2 or 3.

[0048] In some specific embodiments, the compound represented by the above formula IIIa, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, R a , R1 is defined as above, R a The number of is 1, 2 or 3, and one of them is R a It is amino.

[0049] In some specific embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, the formula I has the structure shown in formula IIIa-1:

[0050]

[0051] The compound of formula (IIIa-1) does not include the following compounds

[0052]

[0053] Among them, R a The definition of is as shown above, and n is 1, 2 or 3.

[0054] In some specific embodiments, the compound represented by the above formula IIIa-1, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, R a The definition of R is as shown above. a The number of is 1, 2 or 3, and one of them is R a It is amino.

[0055] In some specific embodiments, the above compound, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, the formula I has the structure shown in formula IIIa-2:

[0056]

[0057] Wherein, the definition of R1 is as shown above.

[0058] As a preferred technical solution of the present invention, the compound of the present invention, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, is selected from the following structures:

[0059]

[0060] Technical terms of the present invention:

[0061] In the present invention, the term "substituted" means that an atom or a group of atoms formally replaces hydrogen and is connected to another group as a "substituent". Unless otherwise noted, the term "substituted" refers to any degree of substitution in the case where such substitution is allowed, such as mono-, di-, tri-, tetra- or penta-substitutions. The substituents are selected independently, and the substitutions can be in any chemically accessible position. It should be understood that the substitution on the specified atom is limited by atomic valence. It should be understood that the substitution on the specified atom produces a chemically stable molecule.

[0062] In the present invention, the term "alkyl" used alone or in combination with other terms refers to a saturated hydrocarbon group which may be a straight chain or a branched chain. 1-8 "Alkyl" refers to an alkyl group having 1 to 8 carbon atoms. Alkyl formally corresponds to an alkane with one CH bond replaced as the point of attachment of the alkyl group to the rest of the compound. In some embodiments, the alkyl group contains 1-8 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl; higher homologues such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like.

[0063] In the present invention, the term "alkoxy" used alone or in combination with other terms refers to a group having the formula -O-alkyl, wherein the term "alkyl" is as defined above. 1-8 "Alkoxy" refers to an alkoxy group whose alkyl group has 1 to 8 carbons. Example alkoxy groups include methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), tert-butoxy, and the like. In some embodiments, the alkyl group has 1 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms.

[0064] In the present invention, the term "halo" or "halogen" used alone or in combination with other terms refers to F, Cl, Br and I. In some embodiments, the term "halo" refers to a halogen atom selected from F, Cl or Br.

[0065] In the present invention, the term "haloalkyl" or "haloalkoxy" used alone or in combination with other terms refers to an alkyl or alkoxy substituted by one or more halogens, wherein the terms "halogen", "alkyl" and "alkoxy" are as defined above.

[0066] In the present invention, the term "deuterated alkyl" used alone or in combination with other terms refers to an alkyl group substituted by one or more deuterium atoms, wherein the term "alkyl" is as defined above.

[0067] In the present invention, the term "aryl" used alone or in combination with other terms refers to an aromatic hydrocarbon group, which may be a monocyclic or polycyclic (eg, having two fused rings). 6-10 "Aryl" refers to an aromatic radical having 6 to 10 ring carbon atoms. Aryl radicals include, for example, phenyl, naphthyl, indanyl, indenyl, and the like. In some embodiments, an aryl radical has 6 carbon atoms. In some embodiments, an aryl radical has 10 carbon atoms. In some embodiments, an aryl radical is phenyl. In some embodiments, an aryl radical is naphthyl.

[0068] In the present invention, the term "heteroaryl", used alone or in combination with other terms, refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from B, P, S, O and N. In some embodiments, the heteroaryl ring has 1, 2, 3 or 4 heteroatom ring members independently selected from S, O and N. In some embodiments, any ring-forming N in the heteroaryl moiety can be an N-oxide. In some embodiments, the heteroaryl has 5 to 14 ring atoms, including carbon atoms and 1, 2, 3 or 4 heteroatom ring members independently selected from S, O and N. In some embodiments, the heteroaryl has 5 to 10 ring atoms, including carbon atoms and 1, 2, 3 or 4 heteroatom ring members independently selected from S, O and N. In some embodiments, the heteroaryl has 5 to 6 ring atoms and 1 or 2 heteroatom ring members independently selected from S, O and N. In some embodiments, the heteroaryl is a five-membered or six-membered heteroaryl ring. In other embodiments, the heteroaryl is an eight-, nine-, or ten-membered fused bicyclic heteroaryl ring. Example heteroaryls include, but are not limited to, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, oxazolyl, oxazolyl, thiazolyl, imidazolyl, furanyl, thienyl, quinolyl, isoquinolyl, naphthyridinyl (including 1,2-naphthyridine, 1,3-naphthyridine, 1,4-naphthyridine, 1,5-naphthyridine, 1,6-naphthyridine, 1,7-naphthyridine, 1,8-naphthyridine, 2,3-naphthyridine, and 2,6-naphthyridine), indolyl, benzothienyl, benzofuranyl, benzisoxazolyl, imidazo[1,2-b]thiazolyl, purinyl, and the like.

[0069] In the present invention, the term "cycloalkyl" used alone or in combination with other terms refers to a saturated ring system (monocyclic, bicyclic or polycyclic), including cyclized alkyl. 3-8 Cycloalkyl" or "C 3-14 "Cycloalkyl" refers to a cycloalkyl group having 3 to 8 or 3 to 14 ring member carbon atoms, respectively. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having 2, 3 or 4 fused rings) groups and spirocyclic and bridged rings. Cycloalkyl groups may have 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring member carbon atoms (C 3-14 ). In some embodiments, the cycloalkyl has 3 to 12 ring members, 3 to 10 ring members, 3 to 8 ring members, 3 to 6 ring members, 3 to 5 ring members, or 3 to 4 ring members. In some embodiments, the cycloalkyl is monocyclic. In some embodiments, the cycloalkyl is monocyclic or bicyclic. In some embodiments, the cycloalkyl is C 3-8Monocyclic cycloalkyl. Cycloalkyl also includes cycloalkylene. In some embodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, norpinenyl, norcarbyl, bicyclo[1.1.1]pentanyl, bicyclo[2.1.1]hexanyl, etc. In some embodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0070] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0071] The chemical reactions of the specific embodiments of the present application are carried out in a suitable solvent, which must be suitable for the chemical changes of the present application and the reagents and materials required. In order to obtain the compounds of the present application, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

[0072] An important consideration in the planning of synthetic routes in this area is to select a suitable protecting group for a reactive functional group (such as an amino group in this application). For example, reference may be made to Chem. Commun., 2019, 55, 7331--7334. All references cited in this application are incorporated herein as a whole. Alternatively, synthetic methods known in the art and the methods described in the present invention are used in combination. The product obtained by each step of the reaction is obtained using separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized according to the literature (such as provided by Scifinder) or purchased.

[0073] In some embodiments, the compounds described herein can be prepared by those skilled in the art of organic synthesis using the following routes and standard methods in the art:

[0074] Preparation of intermediate i

[0075]

[0076] Step 1: The compound represented by formula f and the compound represented by formula g undergo a Grignard reaction to obtain a compound represented by formula h.

[0077] Step 2: The compound represented by formula h undergoes oxidation reaction under the action of an oxidant (such as MnO2) to obtain a compound represented by formula i.

[0078] Synthesis route 1:

[0079]

[0080] Step 1: The compound represented by formula a undergoes a condensation reaction in the presence of a condensation agent (such as EDCI, etc.) to obtain a compound represented by formula b;

[0081] Step 2: Deprotecting the compound represented by formula b under acidic conditions to obtain a compound represented by formula c;

[0082] Step 3: The compound represented by formula c is reacted with ethyl oxalyl chloride under alkaline conditions to obtain the compound represented by formula d;

[0083] Step 4: Under the reducing conditions of the compound represented by formula d, the benzyl ester is hydrolyzed to obtain the compound represented by formula e;

[0084] Step 5: reacting the compound represented by formula e with the compound represented by formula i to obtain a compound represented by formula j;

[0085] Step 6: The compound represented by formula j is hydrolyzed under alkaline conditions at room temperature to obtain the compound represented by formula k;

[0086] Step 7: The compound represented by formula k undergoes a condensation reaction with the compound represented by formula q in the presence of a condensing agent (such as HATU, etc.) to obtain a compound represented by formula l;

[0087] Step 8: The compound represented by formula 1 is heated and hydrolyzed under alkaline conditions to obtain the compound represented by formula m;

[0088] Step 9: reacting the compound represented by formula m with the compound represented by formula p to obtain a compound represented by formula n;

[0089] Step 10: The compound represented by formula n undergoes a coupling reaction with BocNH2 to obtain a compound represented by formula o;

[0090] Step 11: The compound represented by formula o is deprotected under acidic conditions to obtain the compound represented by formula IIIa-2. Synthesis route 2:

[0091]

[0092] Step 1: reacting the compound represented by formula 2a with N-halosuccinimide to obtain the compound represented by formula 2b;

[0093] Step 2: The compound represented by Formula 2b reacts with the compound represented by Formula 2c under alkaline conditions (such as NaH) to obtain the compound represented by Formula 2d;

[0094] Step 3: The compound represented by Formula 2d undergoes an intramolecular coupling reaction to obtain a compound represented by Formula 2e;

[0095] Step 4: The compound represented by Formula 2e undergoes an ester exchange reaction with the compound represented by Formula 2f to obtain a compound represented by Formula 2g;

[0096] Step 5: Deprotecting the compound represented by Formula 2g under acidic conditions to obtain a compound represented by Formula 2h;

[0097] Step 6: Protect the compound represented by Formula 2h under alkaline conditions to obtain the compound represented by Formula 2i;

[0098] Step 7: The compound represented by Formula 2i undergoes Friedel-Crafts reaction with the compound represented by Formula 2j in the presence of a Lewis acid to obtain a compound represented by Formula 2k;

[0099] Step 8: The compound represented by Formula 2k undergoes a condensation reaction with the compound represented by Formula 2l to obtain a compound represented by Formula 2m;

[0100] Step 9: Deprotect the compound represented by Formula 2m under alkaline conditions to obtain the compound represented by Formula II.

[0101] The present invention also provides a pharmaceutical composition comprising a compound of the aforementioned formula (I), (II), (IIa), (IIa-1), (III), (IIIa), (IIIa-1) or (IIIa-2) or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer and pharmaceutically acceptable auxiliary ingredients.

[0102] The present invention also provides the use of the compounds represented by the above formula (I), (II), (IIa), (IIa-1), (III), (IIIa), (IIIa-1) or (IIIa-2) or their pharmaceutically acceptable salts, prodrugs, hydrates, solvates, crystal forms, stereoisomers or tautomers or the above pharmaceutical compositions in the preparation of drugs for preventing and / or improving and / or treating HBV infection.

[0103] Beneficial effects of the present invention:

[0104] The compounds provided by the present invention have good activity in preventing and / or improving and / or treating HBV infection, and can be used as drugs related to the prevention and / or improvement and / or treatment of diseases related to the activity. DETAILED DESCRIPTION

[0105] The present invention is described in detail below by examples, but it is not intended to limit the present invention in any way. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination of the specific embodiments with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art, and preferred embodiments include but are not limited to the embodiments of the present invention. It will be apparent to those skilled in the art that various changes and improvements are made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0106] Example 1 Preparation of Compound 1

[0107]

[0108] Step 1: Synthesis of compound 1-b

[0109] 1-a (7.2 g, 47.06 mmol) was added to the reaction flask, dissolved in carbon tetrachloride (200 mL), and N-bromosuccinimide (8.9 g, 49.88 mmol) was added. Under nitrogen protection, the mixture was stirred at 90°C overnight. Water (200 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (2 x 150 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (PE / EA = 0-15%) to obtain 2.9 g of compound 1-b. MS m / z (ESI): 229.99 [MH] - .

[0110] 1 H NMR (600MHz, DMSO) δ12.22 (s, 1H), 6.09 (d, J = 2.2Hz, 1H), 4.21 (q, J = 7.1Hz, 2H), 2.23 (s, 3H), 1.28 (t, J = 7.1Hz, 3H).

[0111] Step 2: Synthesis of compound 1-d

[0112] Add 1-b (2.9 g, 12.50 mmol) to a two-necked bottle, add N,N-dimethylformamide (80 mL) to dissolve, then cool to 0°C, add sodium hydride (1.3 g, 32.50 mmol) under nitrogen protection, stir in an ice-water bath for 40 minutes, add 1-c (11.2 ml, 50.00 mmol) at 0°C, and then stir at room temperature overnight. Slowly pour the reaction solution into ice water (100 mL), extract the aqueous phase with ethyl acetate (2 x 100 ml), combine the organic layers, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product, which is purified by silica gel column (PE / EA = 5% to 20%) to obtain 3.7 g of compound 1-d. MS m / z (ESI): 375.08 [M+H] + .

[0113] Step 3: Synthesis of compound 1-e

[0114] 1-d (3.7 g, 9.86 mmol) was added to the reaction flask, dissolved in dioxane (150 mL), and then cesium carbonate (9.6 g, 29.58 mmol), methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium (II) (444 mg, 0.49 mmol). Under nitrogen protection, stir at 80 ° C overnight. The reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (PE / EA = 0-20%) to obtain 2.1 g of compound 1-e. MS m / z (ESI): 295.16 [M+H] + .

[0115] Step 4: Synthesis of compound 1-g

[0116] 1-e (2.1 g, 7.11 mmol) was added to the reaction flask, dissolved in dry tetrahydrofuran (80 mL), and then 1-f (1.4 g, 10.84 mmol) was added. Under nitrogen protection, lithium bis(trimethylsilyl)amide (21.33 mL, 21.33 mmol, 1 M) was added dropwise in an ice-water bath. Stir at room temperature for 2 hours. Water (150 mL) was slowly added, and the aqueous phase was extracted with ethyl acetate (2 x 100 ml). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (PE / EA = 10-50%) to obtain 1.7 g of compound 1-g. MS m / z (ESI): 378.16 [M+H] + .

[0117] Step 5: Synthesis of compound 1-h

[0118] Add 1-g (1.7 g, 4.50 mmol) to a reaction flask, dissolve in dichloromethane (30 mL), and then add trifluoroacetic acid (15 mL). Stir at room temperature for 1 hour. Concentrate under reduced pressure to obtain a crude product, and then dissolve the crude product in dichloromethane (40 mL). Adjust the pH of this solution to about 9 with saturated sodium carbonate under an ice-water bath. Add water (50 mL), extract the aqueous phase with dichloromethane (2x 50 mL), combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain 1.1 g of compound 1-h. MS m / z (ESI): 278.10 [M+H] + .

[0119] Step 6: Synthesis of compound 1-i

[0120] 1-h (1.1 g, 3.96 mmol) was added to the reaction flask, dissolved in pyridine (15 mL), and then acetic anhydride (1.12 mL, 11.88 mmol) was added. Stir at room temperature for 1 hour. Water (20 mL) was added, and the aqueous phase was extracted with ethyl acetate (2 x 10 mL). The organic phases were combined, washed with saturated brine, and then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column (PE / EA = 10% to 60%) to obtain 1.1 g of compound 1-i. MS m / z (ESI): 320.11 [M+H] + .

[0121] Step 7: Synthesis of compound 1-k

[0122] 1-i (1.1 g, 3.44 mmol) was added to the reaction flask and dissolved in dry dichloromethane (120 mL). Under nitrogen protection, 1-j (5.5 ml, 51.6 mmol) was first added dropwise in an ice-water bath, and then aluminum chloride (6.9 g, 51.6 mmol) was added. After stirring in an ice-water bath for 1 hour, 1-j (5.5 ml, 51.6 mmol) and aluminum chloride (6.9 g, 51.6 mmol) were added. Stir at room temperature overnight, and water was slowly added dropwise in an ice-water bath to quench the reaction. The aqueous phase (3 x 50 ml) was extracted with a mixed solvent of (DCM / MeOH = 10 / 1), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column (DCM / MeOH = 5% to 60%) to obtain 505 mg of compound 1-k. MS m / z(ESI):392.10[M+H] + .

[0123] Step 8: Synthesis of compound 1-m

[0124] 1-k (39 mg, 0.10 mmol) was added to the reaction flask and dissolved in N, N-dimethylformamide (3 mL). After cooling to 0 ° C, 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (46 mg, 0.12 mmol) and N, N-diisopropylethylamine (70 uL, 0.40 mmol) were added. Stir for 10 minutes in an ice-water bath and 1-l (17 mg, 0.15 mmol) was added. Stir at room temperature for 1 hour. Water (5 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (2 x 10 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 38 mg of crude product compound 1-m. MSm / z (ESI): 487.13 [M+H] + .

[0125] Step 9: Synthesis of compound 1

[0126] 1-m (38 mg, 0.08 mmol) was added to a reaction flask and dissolved in a mixed solvent of methanol (2 mL) and tetrahydrofuran (1 mL). Potassium carbonate (55 mg, 0.40 mmol) was added. Stirred at room temperature for 1 hour. Filtered, the filtrate was purified by high pressure preparation, and freeze-dried to obtain 14 mg of compound 1. MS m / z (ESI): 445.12 [M+H] + .

[0127] 1 HNMR (400MHz, DMSO) δ9.83 (s, 1H), 9.24 (d, J = 9.0Hz, 1H), 7.88–7.66 (m, 1H), 7.48–7.29 (m, 2H), 6.94 (s, 1H), 4.75-4.57 (m, 1H), 4.12 (t, J = 9.0Hz, 2H), 3.90 (t, J = 9.0Hz, 2H), 2.37 (s, 3H), 1.33 (d, J = 7.0Hz, 3H).

[0128] Example 2 Preparation of Compound 2

[0129]

[0130] Step 1: Synthesis of compound 2-b

[0131] Add 1-e (3.0 g, 10.2 mmol) to the reaction flask, dissolve in dry tetrahydrofuran (100 mL), and then add 2-a (2.1 g, 15.3 mmol). Under nitrogen protection, add lithium bis(trimethylsilyl)amide (30.6 mL, 30.6 mmol, 1 M) dropwise in an ice-water bath. Stir at room temperature for 2 hours. Slowly add water (150 mL), extract the aqueous phase with ethyl acetate (2 x 100 mL), combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain a crude product. The crude product is purified by silica gel column (PE / EA = 10-50%) to obtain 2.1 g of compound 2-b. MS m / z (ESI): 385.16 [M+H] + .

[0132] Step 2: Synthesis of compound 2-c

[0133] Add 2-b (2.1 g, 5.45 mmol) to a reaction flask, dissolve in dichloromethane (30 mL), and then add trifluoroacetic acid (15 mL). Stir at room temperature for 1 hour. Concentrate under reduced pressure to obtain a crude product, and then dissolve the crude product in dichloromethane (40 mL). Adjust the pH of the solution to about 9 with saturated sodium carbonate in an ice-water bath. Add water (50 mL), extract the aqueous phase with dichloromethane (2 x 50 mL), combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain 1.4 g of the product, compound 2-c. MS m / z (ESI): 285.11 [M+H] + .

[0134] Step 3: Synthesis of compound 2-d

[0135] Add 2-c (1.4 g, 4.91 mmol) to a reaction flask, dissolve in pyridine (15 mL), and then add acetic anhydride (1.38 mL, 14.73 mmol). Stir at room temperature for 1 hour. Add water (20 mL), extract the aqueous phase with ethyl acetate (2x10 mL), combine the organic phases, add saturated brine to wash, and then dry with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain a crude product. The crude product is purified by silica gel column (PE / EA=10%~60%) to obtain 1.4 g of compound 2-d. MS m / z(ESI):327.12[M+H] + .

[0136] Step 4: Synthesis of compound 2-e

[0137] 2-d (1.4 g, 4.28 mmol) was added to the reaction flask and dissolved in dry dichloromethane (150 mL). Under nitrogen protection, 1-j (6.85 mL, 64.2 mmol) was first added dropwise in an ice-water bath, and then aluminum chloride (8.56 g, 64.2 mmol) was added. After stirring in an ice-water bath for 1 hour, 1-j (6.85 ml, 64.2 mmol) and aluminum chloride (8.56 g, 64.2 mmol) were added. Stir at room temperature overnight, and water was slowly added dropwise in an ice-water bath to quench the reaction. The aqueous phase (3x50 mL) was extracted with a mixed solvent of (DCM / MeOH=10 / 1), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by a silica gel column (DCM / MeOH=5% to 60%) to obtain 612 mg of compound 2-e. MS m / z(ESI):399.10[M+H] + .

[0138] Step 5: Synthesis of compound 2-g

[0139] Add 2-e (40 mg, 0.10 mmol) to the reaction flask and dissolve it in N, N-dimethylformamide (3 mL). After cooling to 0 ° C, add 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (46 mg, 0.12 mmol) and N, N-diisopropylethylamine (70uL, 0.40 mmol). Stir for 10 minutes in an ice-water bath and add 2-f (17 mg, 0.15 mmol). Stir at room temperature for 1 hour. Add water (5 mL) to quench the reaction, extract the aqueous phase with ethyl acetate (2x 10 mL), combine the organic phases, add saturated brine to wash, and then dry with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain 36 mg of crude product compound 2-g. MSm / z (ESI): 494.14 [M+H] + .

[0140] Step 6: Synthesis of Compound 2

[0141] Add 2-g (36 mg, 0.07 mmol) to a reaction flask and dissolve in a mixed solvent of methanol (2 mL) and tetrahydrofuran (1 mL). Add potassium carbonate (48 mg, 0.35 mmol). Stir at room temperature for 1 hour. Filter, purify the filtrate by high pressure preparation, and freeze-dry to obtain 9 mg of compound 2. MS m / z (ESI): 452.13 [M+H] + .

[0142] 1 HNMR (600 MHz, DMSO) δ 9.93 (s, 1H), 9.25 (d, J = 9.0 Hz, 1H), 8.17–8.04 (m, 1H), 8.00–7.89 (m, 1H), 7.51 (t, J = 9.1 Hz, 1H), 6.96 (s, 1H), 4.69–4.61 (m, 1H), 4.18–4.07 (m, 2H), 3.96–3.86 (m, 2H), 2.38 (s, 3H), 1.33 (d, J = 7.1 Hz, 3H). Example 3 Preparation of Compound 3

[0143]

[0144] Step 1: Synthesis of compound 3-a

[0145] The operation steps were the same as those for the synthesis of compound 2-g, except that 2-f was replaced by 1-1 (17 mg, 0.15 mmol), and finally 31 mg of a light yellow solid product, compound 3-a, was obtained. MS m / z (ESI): 494.14 [M+H]+.

[0146] Step 2: Synthesis of compound 3

[0147] The operation steps were the same as those for the synthesis of compound 2, except that 2-g was replaced with 3-a (31 mg, 0.06 mmol), and finally 12 mg of a light yellow solid product, compound 3, was obtained. MS m / z (ESI): 452.13 [M+H]+.

[0148] 1 HNMR(400MHz, DMSO)δ9.93(s,1H),9.24(d,J=8.9Hz,1H),8.12(d,J=3.2Hz,1H),7.97–7.87(m,1H),7.51(t,J=9.1Hz, 1H), 6.96 (s, 1H), 4.71-4.62 (m, 1H), 4.12 (t, J = 8.6Hz, 2H), 3.92 (d, J = 8.7Hz, 2H), 2.38 (s, 3H), 1.33 (d, J = 7.0Hz, 3H).

[0149] Examples 4 and 5 Preparation of Compounds 4 and 5

[0150]

[0151] Step 1: Synthesis of compound 4-b

[0152] 1-e (1.47 g, 4.99 mmol) and 4-a (1.14 g, 5.99 mmol) were added to the reaction flask, dissolved in dry tetrahydrofuran (15 mL), and lithium bis(trimethylsilyl)amide (14.98 mL, 14.98 mmol) was added dropwise under an ice-water bath. Stir at room temperature for 1 hour under nitrogen protection. Slowly add ice water (10 mL) to quench, extract the aqueous phase with ethyl acetate (25 mL), wash the organic phase with saturated brine (20 mL), dry it with anhydrous sodium sulfate, filter it, and concentrate it under reduced pressure to obtain a crude product. The crude product was slurried with petroleum ether (30 mL), filtered, and the filter cake was spin-dried to obtain a white solid compound 4-b (1.67 g, 3.81 mmol). MS m / z (ESI): 438.07 [M+H] + .

[0153] Step 2: Synthesis of compound 4-c

[0154] 4-b (1.67 g, 3.81 mmol) was added to the reaction flask, dissolved in dichloromethane (5 mL), and trifluoroacetic acid (10 mL) was added, and stirred at room temperature for 1 hour. After concentration under reduced pressure, the crude product was dissolved in ethyl acetate (20 mL), and saturated sodium carbonate aqueous solution (20 mL) was added under an ice-water bath, extracted, and the organic phase was washed with saturated brine (30 mL), and then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain an off-white solid compound 4-c (1.29 g, 3.81 mmol). MS m / z (ESI): 338.02 [M+H]+ .

[0155] Step 3: Synthesis of compound 4-d

[0156] Add 4-c (1.29 g, 3.81 mmol) to a reaction flask, add pyridine (10 mL), then add acetic anhydride (1.47 mL, 15.61 mmol), and stir at room temperature for 1 hour. Add water (40 mL), filter, wash the filter cake with water (20 mL), and dry to obtain an off-white solid compound 4-d (1.2 g, 3.16 mmol). MS m / z (ESI): 382.05 [M+H] + .

[0157] Step 4: Synthesis of compound 4-e

[0158] 4-d (1.2 g, 3.16 mmol) was added to the reaction flask, and dichloromethane (15 mL) was added. Ethyl oxalyl chloride (12.36 mL, 110.47 mmol) was added under an ice-water bath, and aluminum chloride (14.7 g, 110.47 mmol) was added. Stir at room temperature overnight under nitrogen protection. Ice water (50 mL) was slowly added dropwise under an ice-water bath to quench. The aqueous phase was extracted with ethyl acetate (50 mL), and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain an orange solid compound 4-e (1.0 g, 2.21 mmol). MS m / z (ESI): 452.06 [M+H] + .

[0159] Step 5: Synthesis of compound 4-f

[0160] 4-e (1.0 g, 2.21 mmol) and 2-f (397 mg, 2.65 mmol) were added to a reaction flask, dissolved in DMF (20 mL), and N, N-diisopropylethylamine (1.47 mL, 8.85 mmol) was added. The reaction flask was placed in an ice-water bath and stirred, and then HATU (1.26 g, 3.32 mmol) was added and stirred in an ice-water bath for 20 minutes. The reaction solution was directly prepared and purified (eluent ratio: acetonitrile / 0.05% formic acid aqueous solution = 65% / 35%) to obtain an orange solid compound 4-f (0.90 g, 1.62 mmol). MS m / z (ESI): 549.02 [M+H] + .

[0161] Step 6: Synthesis of compound 5

[0162] 4-f (679 mg, 1.24 mmol) was added to the reaction flask, dissolved in a mixed solvent of methanol (20 mL) and tetrahydrofuran (20 mL), potassium carbonate (857 mg, 6.20 mmol) was added, and stirred at room temperature for 1 hour. Filter, concentrate the filtrate, prepare and purify, and freeze-dry to obtain yellow solid compound 5 (600 mg, 1.19 mmol). MS m / z (ESI): 507.09 [M+H] + .

[0163] 1 H NMR (400MHz, DMSO-d6) δ9.78 (s, 1H), 9.25 (d, J = 9.0Hz, 1H), 8.09-8.03 (m, 1H), 7.67-7.61 (m, 1H), 7.36 (t, J = 8.8Hz, 1 H), 6.95 (s, 1H), 4.80–4.64 (m, 1H), 4.14 (t, J = 8.7Hz, 2H), 3.93 (t, J = 8.8Hz, 2H), 2.40 (s, 3H), 1.35 (d, J = 7.0Hz, 3H).

[0164] Step 7: Synthesis of compound 4-g

[0165] Compound 5 (180 mg, 0.36 mmol) was added to a reaction flask, dissolved in acetonitrile (10 mL), and di-tert-butyl dicarbonate (0.41 mL, 1.80 mmol), triethylamine (0.15 mL, 1.08 mmol) and DMAP (43 mg, 0.36 mmol) were added, and stirred at room temperature overnight. The reaction solution was concentrated and purified by silica gel column to obtain an orange solid compound 4-g (140 mg, 0.20 mmol). MS m / z (ESI): 705.2 [M+H] + .

[0166] Step 8: Synthesis of compound 4-h

[0167] 4-g (140 mg, 0.20 mmol) was added to a sealed tube, followed by tert-butyl carbamate (232 mg, 1.98 mmol), cesium carbonate (194 mg, 0.60 mmol) and BrettPhos Pd G3 (27 mg, 0.03 mmol), 1,4-dioxane (2 mL) was added, nitrogen was replaced, the tube was sealed, heated to 100°C, and stirred overnight. Concentrated, purified by silica gel column (PE / EA=3 / 1), and yellow solid compound 4-h (60 mg, 0.08 mmol) was obtained. MS m / z (ESI): 742.4 [M+H] + .

[0168] Step 9: Synthesis of compound 4

[0169] 4-h (60 mg, 0.08 mmol) was added to the reaction flask, dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added, and stirred at room temperature for 1 hour. The reaction solution was concentrated and purified by C18 column, eluent ratio: 55% (acetonitrile): 45% (0.05% formic acid aqueous solution), the purified product was collected and lyophilized to obtain yellow solid compound 4 (18 mg, 0.04 mmol). MS m / z (ESI): 442.17 [M+H] + .

[0170] 1 H NMR (400MHz, DMSO-d6) δ9.39 (s, 1H), 9.20 (d, J = 9.0Hz, 1H), 7.17 (dd, J = 8.4, 2.6Hz, 1H), 6.92-6.86 (m, 2H), 6.76-6.70 ( m,1H),5.11(s,2H),4.73–4.59(m,1H),4.10(t,J=8.6Hz,2H),3.90(t,J=8.6Hz,2H),2.35(s,3H),1.33(d,J=7.0Hz,3H).

[0171] Example 6 Preparation of Compound 6

[0172]

[0173] Step 1: Synthesis of compound 6-b

[0174] The operation steps were the same as those for the synthesis of compound 4-b, except that 4-a was replaced with 6-a (3.6 g 24.5 mmol), and 4 g of white solid product compound 6-b was finally obtained. MS m / z (ESI): 396.15 [M+H] + .

[0175] Step 2: Synthesis of compound 6-c

[0176] The operation steps were the same as those for the synthesis of compound 4-c, except that 4-b was replaced with 6-b (4 g, 10.1 mmol), and 2.8 g of product compound 6-c was finally obtained. MS m / z (ESI): 296.09 [M+H] + .

[0177] Step 3: Synthesis of compound 6-d

[0178] The operation steps were the same as those for the synthesis of compound 4-d, except that 4-c was replaced with 6-c (2.8 g, 9.5 mmol), and 2.4 g of the product compound 6-d was obtained as a white solid. MS m / z (ESI): 338.10 [M+H] + .

[0179] Step 4: Synthesis of compound 6-e

[0180] The operation steps were the same as those for the synthesis of compound 4-e, except that 4-d was replaced with 6-d (2.4 g, 7.1 mmol), and 1.3 g of light yellow solid product compound 6-e was finally obtained. MS m / z (ESI): 410.09 [M+H] + .

[0181] Step 5: Synthesis of compound 6-f

[0182] The operation steps were the same as those for the synthesis of compound 4-f, except that 4-e was replaced with 6-e (100 mg, 0.17 mmol), and 80 mg of a light yellow solid product compound 6-f was finally obtained. MS m / z (ESI): 505.11 [M+H] + .

[0183] Step 6: Synthesis of compound 6

[0184] The operation steps were the same as those for the synthesis of compound 5, except that 4-f was replaced with 6-f (80 mg, 0.16 mmol), and finally 45 mg of yellow solid product compound 6 was obtained. MS m / z (ESI): 463.04 [M+H] + .

[0185] 1 H NMR (400MHz, DMSO) δ9.94 (s, 1H), 9.26 (d, J = 9.0Hz, 1H), 7.68–7.51 (m, 2H), 6.98 (s, 1H), 4. 73-4.65(m,1H),4.17-4.11(m,2H),4.03-3.87(m,2H),2.39(s,3H),1.35(d,J=7.0Hz,3H).

[0186] Example 7 Preparation of Compound 7

[0187]

[0188] Step 1: Synthesis of compound 7-a

[0189] The operation steps were the same as those for the synthesis of compound 6-f, except that 2-f was replaced with 1-1 (44 mg, 0.29 mmol), and finally 80 mg of light yellow solid product compound 7-a was obtained. MS m / z (ESI): 505.11 [M+H] + .

[0190] Step 2: Synthesis of compound 7

[0191] The operation steps were the same as those for the synthesis of compound 6, except that 6-f was replaced with 7-a (80 mg, 0.16 mmol), and finally 45 mg of a light yellow solid product, compound 7, was obtained. MS m / z (ESI): 463.07 [M+H] + .

[0192] 1 H NMR (400MHz, DMSO) δ9.94 (s, 1H), 9.26 (d, J = 8.8Hz, 1H), 7.65–7.53 (m, 2H), 6.98 (s, 1H), 4.73-4. 65(m,1H),4.19–4.10(m,2H),3.98–3.88(m,2H),3.31(s,2H),2.39(s,3H),1.35(d,J=7.0Hz,3H).

[0193] Example 8 Preparation of Compound 8

[0194]

[0195] Step 1: Synthesis of compound 8-b

[0196] 8-a (20 g, 92.94 mmol) was added to a reaction flask, dissolved in dichloromethane (200 mL), and benzyl alcohol (14.42 mL, 139.41 mmol), EDCI (35.6 g, 185.87 mmol), N,N-diisopropylethylamine (61.58 mL, 371.75 mmol) and DMAP (2.3 g, 18.59 mmol) were added, and stirred at room temperature overnight. The reaction solution was washed three times with 5% citric acid aqueous solution (200 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an oily liquid compound 8-b (24.4 g, 79.9 mmol). MS m / z (ESI): 250.10 [M-56+H] + .

[0197] Step 2: Synthesis of compound 8-c

[0198] 8-b (24.4 g, 79.9 mmol) was added to the reaction flask, HCl / EA (200 mL, 400 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the crude product was slurried with PE (200 mL), filtered, and dried to obtain the hydrochloride salt of white solid compound 8-c (15.9 g, 65.77 mmol). MS m / z (ESI): 206.14 [M+H] + .

[0199] Step 3: Synthesis of compound 8-d

[0200] The hydrochloride of 8-c (15.9 g, 65.77 mmol) was added to the reaction flask, dissolved in dichloromethane (200 mL), and N, N-diisopropylethylamine (21.78 mL, 371.75 mmol) was added. In an ice-water bath, ethyl oxalyl chloride (11.04 mL, 98.64 mmol) was slowly added. Under nitrogen protection, the mixture was stirred at room temperature for 1 hour. The reaction solution was placed in an ice-water bath, quenched with saturated sodium bicarbonate aqueous solution (50 mL), extracted with dichloromethane (50 mL), and the organic phases were combined. The organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an orange solid compound 8-d (20 g, 65.51 mmol). MS m / z (ESI): 306.15 [M+H] + .

[0201] Step 4: Synthesis of compound 8-e

[0202] 8-d (10.6 g, 34.72 mmol) was added to the reaction flask, dissolved in ethanol (100 mL), and Pd / C (2.6 g, 10 wt%, containing 50% water) was added. The mixture was stirred at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered and the filtrate was concentrated to obtain an oily liquid compound 8-e (7.0 g, 32.53 mmol). MS m / z (ESI): 216.12 [M+H] + .

[0203] Step 5: Synthesis of compound 8-h

[0204] 8-f (1.67 g, 3.81 mmol) was added to the reaction flask, dissolved in toluene (23 mL), and nitrogen was protected. At -40°C, 8-g (100 mL, 50 mmol) was added dropwise, and stirred at -40°C for 1.5 hours. The temperature was slowly raised to 0°C, quenched by adding saturated aqueous ammonium chloride solution (20 mL), extracted twice with ethyl acetate (20 mL), and the organic phases were combined, washed with saturated brine (30 mL), and then dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified on a silica gel column to obtain an orange-yellow oily liquid compound 8-h (1.0 g, 7.03 mmol). 1 H NMR (400MHz, DMSO) δ6.07 (s, 1H), 4.78 (s, 1H), 4.19-4.13 (m, 2H), 1.84 (d, J = 2.4Hz, 3H), 1.23 (t, J = 7.0Hz, 3H). 1 H NMR (400MHz, CDCl3) δ4.36 (q, J = 7.2Hz, 2H), 2.15 (s, 3H), 1.39 (t, J = 7.1Hz, 3H).

[0205] Step 6: Synthesis of compound 8-i

[0206] 8-h (1.0 g, 7.03 mmol) was added to the reaction flask, dissolved in dichloromethane (20 mL), and manganese dioxide (2.44 g, 28.13 mmol) was added. Under nitrogen protection, the mixture was stirred at room temperature overnight. The reaction solution was filtered and the filtrate was concentrated to obtain a light yellow oily liquid compound 8-i (980 mg, 7.0 mmol). 1 H NMR (400MHz, CDCl3) δ4.36 (q, J = 7.2Hz, 2H), 2.15 (s, 3H), 1.39 (t, J = 7.1Hz, 3H).

[0207] Step 7: Synthesis of compound 8-j

[0208] 8-e (1.0 g, 4.65 mmol) and 8-i (788 mg, 5.11 mmol) were added to a microwave tube, dissolved in N-methylpyrrolidone (10 mL), and N, N'-diisopropylcarbodiimide (680 mg, 5.39 mmol) was added. The mixture was replaced with nitrogen, sealed, and heated to 140 ° C in a microwave oven for 40 minutes. Saturated aqueous ammonium chloride solution (2 mL) was added to quench, and ethyl acetate (5 mL) was used to extract twice. The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified on a silica gel column to obtain a light yellow oily liquid compound 8-j (100 mg, 0.34 mmol). MS m / z (ESI): 294.17 [M+H] + .

[0209] Step 8: Synthesis of compound 8-k

[0210] 8-j (100 mg, 0.34 mmol) was added to the reaction flask, dissolved in a mixed solvent of ethanol (2 mL) and water (0.2 mL), sodium hydroxide (27 mg, 0.68 mmol) was added under an ice-water bath, and stirred at room temperature for 1 hour. The pH was adjusted to 5, concentrated under reduced pressure, prepared and purified, and freeze-dried to obtain a white solid compound 8-k (82 mg, 0.31 mmol). MS m / z (ESI): 266.13 [M+H] + .

[0211] Step 9: Synthesis of compound 8-1

[0212] The operation steps were the same as those for the synthesis of compound 4-f, except that 4-e was replaced with 8-k (82 mg, 0.31 mmol), and finally a light yellow solid product, compound 8-1 (75 mg, 0.21 mmol) was obtained. MS m / z (ESI): 361.14 [M+H] + . 1H NMR(400MHz,DMSO)δ9.31(d,J=8.8Hz,1H),4.76-4.67(m,1H),4.36–4.21(m ,4H),2.93-2.89(m,2H),2.55(s,3H),2.49–2.40(m,2H),1.34-1.31(m,6H).

[0213] Step 10: Synthesis of compound 8-m

[0214] 8-1 (70 mg, 0.19 mmol) was added to the reaction flask, dissolved in a mixed solvent of ethanol (3 mL) and water (0.6 mL), sodium hydroxide (152 mg, 3.8 mmol) was added, heated to 60°C, and stirred for 2 hours. The pH was adjusted to 5, concentrated under reduced pressure, purified, and freeze-dried to obtain a white solid compound 8-m (63 mg, 0.19 mmol). MS m / z (ESI): 333.24 [M+H] + .

[0215] Step 11: Synthesis of compound 8-n

[0216] 8-m (63 mg, 0.19 mmol) was added to the reaction flask, dissolved in thionyl chloride (5 mL), heated to 50 ° C under nitrogen protection, and stirred for 20 minutes. The reaction solution was concentrated, dissolved in tetrahydrofuran (5 mL), 4-a (180 mg, 0.95 mmol) was added, and stirred for 2 seconds. The reaction solution was concentrated and purified to obtain an orange solid compound 8-n (90 mg, 0.18 mmol). MS m / z (ESI): 504.03 [M+H] + .

[0217] Step 12: Synthesis of compound 8-o

[0218] The operation steps were the same as those for the synthesis of compound 4-h, except that 4-g was replaced with 8-n (90 mg, 0.18 mmol), and finally a light yellow solid product compound 8-o (90 mg, 0.17 mmol) was obtained. MS m / z (ESI): 361.14 [M+H] + .

[0219] Step 13: Synthesis of compound 8

[0220] The operation steps were the same as those for the synthesis of compound 4, except that 4-h was replaced with 8-o (90 mg, 0.17 mmol), and a white solid product, compound 8 (40 mg, 0.09 mmol), was finally obtained. MS m / z (ESI): 441.25 [M+H] + .

[0221] 1H NMR (400MHz, DMSO) δ9.59(s,1H),9.26(d,J=8.8Hz,1H),7.19(dd,J=8.4,2.6Hz,1H),6.93-6.88(m,1H),6.77-6.73(m,1 H),5.15(s,2H),4.72-4.67(m,1H),4.13(t,J=7.2Hz,2H),2.93–2.84(m,2H),2.46-2.39(m,5H),1.32(d,J=7.0Hz,3H).

[0222] Example 9 Preparation of Compound 9

[0223]

[0224] Step 1: Synthesis of compound 9-a

[0225] The operation steps were the same as those for the synthesis of compound 1-m, except that 1-1 was replaced with 2-f (17 mg, 0.15 mmol), and finally 29 mg of light yellow solid product compound 9-a was obtained. MS m / z (ESI): 487.13 [M+H] + .

[0226] Step 2: Synthesis of compound 9

[0227] The operation steps were the same as those for the synthesis of compound 1, except that 1-m was replaced with 9-a (29 mg, 0.06 mmol), and finally 10 mg of light yellow solid product compound 9 was obtained. MS m / z (ESI): 445.12 [M+H] + .

[0228] 1 HNMR(600MHz,DMSO)δ9.82(s,1H),9.24(d,J=9.0Hz,1H),7.79(dd,J=12.8,8.0Hz,1H),7.43–7.35(m,2H),6 .94(s,1H),4.70–4.64(m,1H),4.12(t,J=9.3Hz,2H),3.96–3.80(m,2H),2.37(s,3H),1.33(d,J=7.1Hz,3H).

[0229] Example: In vitro anti-HBV activity experiment of HepG2.2.15 cells

[0230] 1. Compound dilution: dilute the 20 μM compound stock solution 10-fold with DMSO to obtain a 2 μM concentration of the compound, and then dilute the 20 μM and 2 μM compounds 200-fold with culture medium to obtain compound solutions with final concentrations of 100 nM and 10 nM, respectively.

[0231] 2. Experimental operation: On the first day, HepG2.2.15 cells (Wuhan Institute of Virology, Chinese Academy of Sciences) (1×10 5 Cells / well) were plated into a 96-well cell culture plate and cultured overnight at 37°C, 5% CO2. Wells without cells and containing only culture medium were set as culture medium controls. On the second day, culture medium with different final concentrations of compounds in step 1 was added to the cell culture plate, and 3 replicate wells were set for each concentration. The final concentration of DMSO in the cell culture medium was 0.5%, and the final volume in the culture wells was 200 μl. Wells without compounds with a final concentration of 0.5% DMSO were set as DMSO controls. Culture was continued until the fifth day, and fresh culture medium with the same treatment as above was replaced in the compound-treated wells and the control wells. On the eighth day, the cell supernatant in the culture wells was collected, and part of the sample was taken to detect the HBV DNA content. A plasmid containing the full-length sequence of type D HBV DNA was used as a qPCR standard, and the standard range was 10 7 -10 1 100 copies / μl, qPCR was used to determine the content of HBV DNA in all samples. After collecting the supernatant, Cell Titer-Glo reagent was added to the cell culture wells, incubated at room temperature for 10 minutes, and then the chemiluminescence values ​​in all wells were read by a microplate reader to calculate the cell proliferation activity.

[0232] 3. Data analysis: The inhibition percentage was calculated using the following formula:

[0233] HBV DNA inhibition rate %=(1-DNA copy number in compound sample / DNA copy number in DMSO control)×100%.

[0234] Cell Viability % = (luminescence value in compound sample - luminescence value of medium control) / (luminescence value of DMSO control - luminescence value of medium control) x 100%.

[0235] The compounds tested according to the above dilution method were used to obtain the anti-HBV replication activity EC value according to the HBV DNA inhibition rate (%). 50 scope.

[0236] Using the above experimental method, the anti-HBV activity and corresponding cell proliferation inhibition activity of some compounds of the present invention are shown in Table 1 below.

[0237] Table 1. HepG2.2.15 cell HBV DNA inhibition

[0238]

[0239] It can be seen from Table 1 that the compounds of the present invention have excellent anti-HBV activity.

[0240] Industrial Applicability

[0241] The compounds of the present invention have excellent anti-HBV activity and can be used as drugs for treating or preventing diseases related to the activity.

[0242] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

[0243] References:

[0244] [1]Villain P, Gonzalez P, Almonte M, et al. European Code against Cancer4th Edition: Infections and Cancer[J]. Cancer Epidemiology, 2015.

[0245] [2]Samuel,E C.Antiviral actions of interferons[Table of Contents][J].Clinical Microbiology Reviews, 2001,14(4):778-809.

[0246] [3]ARGG,BBG,CCLL,et al.Chronic hepatitis B: Virology,natural history,current management and a glimpse at future opportunities-ScienceDirect[J].Antiviral Research,2015,121:47-58.

[0247] [4] Elias S, Smith CI, Ghany M G. Hepatitis B: Current Status of Therapy and Future Therapies[J]. Gastroenterology clinics of North America, 2021.

[0248] [5]Berke,Martin J,Dehertogh,et al.Capsid assembly modulator JNJ-56136379prevents de novo infection of primary human hepatocytes withhepatitis B virus.

[0249] [6]Berke J M,Dehertogh P,Vergauwen K,et al.Capsid Assembly ModulatorsHave a Dual Mechanism of Action in Primary Human Hepatocytes Infected withHepatitis B Virus[J].Antimicrob Agents Chemother,2017:AAC.00560-17.

[0250] [7]Angela,M,Lam,et al.Preclinical Characterization of NVR 3-778,aFirst-in-Class Capsid Assembly Modulator Against the Hepatitis B Virus.[J].Antimicrobial Agents&Chemotherapy,2018。

Claims

1. A compound represented by formula (I), or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof: in, X1 is selected from -NH- or -CH2-; R1 is selected from -H, deuterium, C 1-8 Alkyl, C 3-8 Cycloalkyl, C 1-8 Haloalkyl, C 1-8 Deuterated alkyl, C 1-8 Alkoxy, C 1-8 Deuterated alkoxy, halogen, -CN, -OH, -NH2; R2 is selected from unsubstituted or substituted by one or more R a Substituted C 6-10 Aryl, unsubstituted or substituted with one or more R b Substituted 5-10 membered heteroaryl; the 5-10 membered heteroaryl contains 1-3 heteroatoms selected from N, O and S; Each R a , R b are independently selected from the same or different halogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 1-8 Haloalkoxy, -CN, -NH2; The compound of formula (I) does not include the following compounds 2. The compound according to claim 1, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: R1 is selected from -H, deuterium, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Deuterated alkyl, C 3-6 Cycloalkyl, halogen, -CN, -OH, -NH2; Preferably, R1 is selected from C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Deuterated alkyl, halogen, -CN, -NH2; Preferably, R1 is selected from C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Deuterated alkyl, halogen, -CN, -NH2; Preferably, R1 is selected from C 1-2 Alkyl, C 3-4 Alkyl, C 1-2 Fluorinated alkyl, halogen; Preferably, R1 is selected from methyl, ethyl, trifluoromethyl, -Cl, -Br; Most preferably, R1 is selected from methyl.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted 5-6 membered heteroaryl; the 5-6 membered heteroaryl contains 1-3 heteroatoms selected from N, O and S; each R a , R b are independently selected from the same or different halogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 1-8 Haloalkoxy, -CN, -NH2; Preferably, R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted pyridyl or pyrimidinyl; each R a , R b are independently selected from the same or different halogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 1-8 Haloalkoxy, -CN, -NH2; Preferably, R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted pyridyl or pyrimidinyl; each R a , R b are independently selected from the same or different halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, -CN, -NH2; Preferably, R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl, unsubstituted or replaced by one or more R b Substituted pyridyl or pyrimidinyl; each R a , R b are independently selected from the same or different halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, -CN, -NH2; Preferably, R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl; each R a are independently selected from the same or different halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, -CN, -NH2; Preferably, R2 is selected from unsubstituted or substituted by one or more R a Substituted phenyl; each R a Each is independently selected from the same or different F, Cl, Br, methyl, ethyl, isopropyl, fluoromethyl, fluoroethyl, fluoromethoxy, fluoroethoxy, -CN, -NH2; Preferably, R2 is selected from unsubstituted or substituted with 1, 2 or 3 R a Substituted phenyl; each R a Each independently selected from the same or different F, Cl, Br, -CN, -NH2; Most preferably, R2 is selected from 4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: Formula I is selected from the structure shown in Formula II: Wherein, the definitions of R1 and R2 are as shown in any one of claims 1-3; Further, Formula II is selected from the structure shown in Formula IIa: Among them, R1, R a As defined in any one of claims 1 to 3, m is 1, 2 or 3; Furthermore, Formula IIa is selected from the structure shown in Formula IIa-1: Among them, R a As defined in any one of claims 1 to 3, m is 1, 2 or 3.

5. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: Formula I is selected from the structure shown in Formula III: The compound of formula (III) does not include the following compounds Wherein, the definitions of R1 and R2 are as shown in any one of claims 1-3; Further, Formula III is selected from the structure shown in Formula IIIa: The compound of formula (IIIa) does not include the following compounds Among them, R a , R1 is defined as shown in any one of claims 1-3, and n is 1, 2 or 3; Further, Formula IIIa is selected from the structure shown in Formula IIIa-1 or IIIa-2: The compound of formula (IIIa-1) does not include the following compounds Among them, R a As defined in any one of claims 1 to 3, n is 1, 2 or 3; Wherein, the definition of R1 is as shown in any one of claims 1-3.

6. The compound according to claim 5, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: In Formula IIIa, R a The number of is 1, 2 or 3, and one of them is R a It is amino.

7. The compound according to claim 5, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: In Formula IIIa-1, R a The number of is 1, 2 or 3, and one of them is R a It is amino.

8. The compound according to claim 1, or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof, characterized in that: The compound is selected from the following structures:

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer thereof and a pharmaceutically acceptable auxiliary ingredient.

10. Use of the compound according to any one of claims 1 to 8 or its pharmaceutically acceptable salt, prodrug, hydrate, solvate, crystal form, stereoisomer or tautomer or the pharmaceutical composition according to claim 9 in the preparation of a medicament for preventing and / or improving and / or treating HBV infection.

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