Dengue virus replication inhibitor, preparation method therefor and use thereof

Abstract

The present invention provides a novel small molecule compound having Dengue virus replication inhibitory activity, relates to a compound represented by formula (II), a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, a preparation method therefor, and the use of the compound, and belongs to the field of medicinal chemistry.

Description

Dengue virus replication inhibitors, methods of making and uses thereof TECHNICAL FIELD

[0001] The present application belongs to the field of medicinal chemistry, and particularly relates to a class of dengue virus replication inhibitors, methods of making and uses thereof. BACKGROUND

[0002] Dengue fever (also known as bone pain fever) is caused by dengue virus infection. The severe form of the disease, dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), has a fatal risk to children. Dengue virus (DENV) belongs to the Flavivirus genus of the Flaviviridae family, and other viruses of this family include ZIKA, Japanese encephalitis virus (JEV), West Nile virus (WNV) and yellow fever virus (YFV). The currently prevalent dengue virus is mainly divided into 4 serotypes, DENV-1, DENV-2, DENV-3, DENV-4, and each serotype of dengue virus repeatedly dominates the outbreak of dengue fever in different times and different regions. Infection with different serotypes of dengue virus can have a significant antibody-dependent enhancement effect (ADE effect), and the antibodies obtained after recovery from infection with one serotype cannot effectively neutralize another serotype of dengue virus in the second infection, but instead promote the entry of the second infection dengue virus into cells and accelerate the replication of the virus in the body, especially in children and immunocompromised patients, leading to more patients developing into severe cases, showing dengue hemorrhagic fever and dengue shock syndrome, and even death. The current multivalent dengue virus vaccine shows uneven resistance to multiple serotypes, so it is necessary to develop specific drugs against dengue virus to achieve broad-spectrum resistance to dengue virus and avoid the occurrence of ADE effect. SUMMARY

[0003] The first aspect of the present application relates to a compound represented by formula (II), a stereoisomer, a tautomer thereof or a pharmaceutically acceptable salt thereof,

[0004] wherein ring A is a tricyclic fused ring, the fused ring containing one or more heteroatoms;

[0005] X is H or halogen;

[0006] R 1 each independently is -OC 1-3 alkyl, the -OC 1-3 alkyl is optionally substituted with one or more halogens;

[0007] R 2 is H or -OC 1-3 alkyl;

[0008] R 3 H or -OC 1-3 alkyl;

[0009] R 4 H or -S(=O)2C 1-3 alkyl;

[0010] n is 0, 1, 2 or 3.

[0011] In some embodiments of the present application, the heteroatom contained in the fused ring is N.

[0012] In some embodiments of the present application, one heteroatom is contained in the fused ring.

[0013] In some embodiments of the present application, the ring A is a tricyclic fused aromatic heterocycle, and the heteroatom of the tricyclic fused aromatic heterocycle is N; preferably, the number of the heteroatom is 1.

[0014] In some embodiments of the present application, the ring A comprises a structure of benzo-penta-azacyclic ring, and the structure of benzo-penta-azacyclic ring is preferably

[0015] In some embodiments of the present application, the ring A is wherein the ring B is a 4-7-membered heterocycloalkane, a 4-7-membered cycloalkane, a 4-7-membered aromatic ring or a 4-7-membered aromatic heterocycle.

[0016] In some embodiments of the present application, the ring A is preferably

[0017] In some embodiments of the present application, is preferably

[0018] In some embodiments of the present application, the ring B is a 5-6-membered heterocycloalkane, a 5-6-membered cycloalkane, a 5-6-membered aromatic ring or a 5-6-membered aromatic heterocycle.

[0019] In some embodiments of the present application, the compound represented by the formula (II), a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, has any of the following structures:

[0020] wherein n is each independently 0, 1, 2 or 3. 1 the substitutable position of the tricyclic fused ring can be optionally substituted;

[0021] Preferably, the compound of formula (II), stereoisomer, tautomer or pharmaceutically acceptable salt thereof has a structure of formula (II-4).

[0022] In some embodiments of the present application, R 1 is -OC 1-3 alkyl, said -OC 1-3 alkyl being optionally substituted with one or more F; preferably, R 1 is -OCH3, -OCHF2, -OCH2F or -OCF3, more preferably -OCF3.

[0023] In some embodiments of the present application, n is 0, 1 or 2, preferably 0 or 1.

[0024] In some embodiments of the present application, X is halogen (e.g. F, CI or Br), preferably -CI.

[0025] In some embodiments of the present application, R 2 is -OC 1-3 alkyl, preferably -OCH3.

[0026] In some embodiments of the present application, R 3 is -OC 1-3 alkyl, preferably -OCH3.

[0027] In some embodiments of the present application, R 4 is -S(=O)2C 1-3 alkyl, preferably -S(=O)2CH3.

[0028] In some embodiments of the present application, the compound of formula (II), stereoisomer, tautomer or pharmaceutically acceptable salt thereof has a structure of any one of the following:

[0029] In some embodiments of the present application, the compound of formula (II), stereoisomer, tautomer or pharmaceutically acceptable salt thereof has a structure of any one of the following:

[0030] The second aspect of the present application provides a method for preparing a compound of formula (II), stereoisomer, tautomer or pharmaceutically acceptable salt thereof, comprising the following steps:

[0031] wherein X, R 1 , R 2 , R 3 , R 4 and n are each independently described in the first aspect of the present application;

[0032] Step 1: converting intermediate a1 into intermediate a2 in the presence of chlorinating reagent;

[0033] The chlorinating reagent includes but is not limited to SOCl2, POCl3, (COCl)2;

[0034] Step 2: converting intermediate a3 and intermediate a2 into intermediate a4 in the presence of Lewis acid in a suitable solvent;

[0035] The suitable solvent in Step 2 includes but is not limited to DCM, 1,2-dichloroethane;

[0036] The Lewis acid includes but is not limited to Et2AlCl, TiCl4;

[0037] Step 3: converting intermediate a4 into intermediate a5 in the presence of brominating reagent in a suitable solvent;

[0038] The suitable solvent in Step 3 includes but is not limited to THF, dioxane;

[0039] The brominating reagent includes but is not limited to CuBr2, phenyltrimethylammonium tribromide;

[0040] Step 4: converting intermediate a5 into the compound of formula (II) in the presence of base in a suitable solvent;

[0041] The suitable solvent in Step 4 includes but is not limited to acetonitrile;

[0042] The base includes but is not limited to DIEA, TEA;

[0043] The compound of formula (II) can be obtained by chiral resolution to obtain corresponding isomers IIa and IIb;

[0044] The chiral resolution includes resolution by SFC.

[0045] The third aspect of the present application provides a pharmaceutical composition comprising at least one compound of formula (II), stereoisomer, tautomer thereof or pharmaceutically acceptable salt thereof of the present application, and optionally one or more pharmaceutically acceptable carriers and / or additives.

[0046] The fourth aspect of the present application provides use of a compound of formula (II), stereoisomer, tautomer thereof or pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present application in the preparation of a medicament for preventing or treating flavivirus infection; preferably, the flavivirus infection is dengue virus or yellow fever virus infection.

[0047] The fifth aspect of the present invention provides the use of a compound of formula (II), its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition of the present invention, in the preparation of a medicament for inhibiting the replication of one or more flaviviruses in a biological sample or an individual in need.

[0048] The sixth aspect of the present invention provides a method for preventing or treating flavivirus infection in an individual in need, the method comprising administering to the individual a therapeutically effective amount of a compound of formula (II), its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition of the present invention.

[0049] The flaviviruses described in this invention include, but are not limited to, dengue virus, Zika virus, yellow fever virus, and Japanese encephalitis virus; preferably, the flavivirus is dengue virus. Attached Figure Description

[0050] Figure 1: Anti-dengue virus type 2 cell efficacy of compound 5 and JNJ-1802 (EC5) 50 ) curve, Vero cytotoxicity (CC) 50 The curve and the selection index (SI);

[0051] Figure 2: Efficacy evaluation of compound 5 and JNJ-1802 in DENV2-infected AG129 mouse model (A: Experimental procedure; B: Mouse survival rate; C: Plasma viral load on day 3; D: Plasma viral load on days 10 / 15 / 20). Detailed Implementation

[0052] Unless otherwise stated, conventional methods within the scope of the art, such as mass spectrometry, NMR, and pharmacological methods, are employed. Unless specifically defined, the terminology used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, reactions and purifications can be carried out using the manufacturer's instructions for use of reagent kits, or in accordance with methods known in the art or the descriptions of this invention. The techniques and methods described herein are generally carried out according to conventional methods well known in the art, based on the descriptions in the various summary and more specific literatures cited and discussed herein.

[0053] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0054] The term "optionally" includes both "optional" and "not optional". For example, "optionally substituted methyl" means "unsubstituted methyl" or "substituted methyl".

[0055] The term "substituted" means that any one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded. When a substituent is oxo or keto (i.e., =0), then 2 hydrogens on the atom are replaced.

[0056] "or" means that the listed alternatives are to be taken one at a time, and that the interposed term "or" is not intended to imply alternatives from among its own terms. "and" means that the listed alternatives are to be taken together. "or" means that the listed alternatives are to be taken one at a time, and that the interposed term "or" is not intended to imply alternatives from among its own terms.

[0057] The term "fused ring" means two or more carbocyclic or heterocyclic rings sharing a ring bond. The carbocyclic or heterocyclic rings can be optionally saturated or unsaturated rings, and can optionally form aromatic rings. There can be one or more bonds shared between the individual rings of the fused ring.

[0058] The term "tricyclic fused ring" means three carbocyclic or heterocyclic rings sharing a ring bond. For example, a fused ring of three carbocyclic rings, a fused ring of two carbocyclic rings and one heterocyclic ring, a fused ring of one carbocyclic ring and two heterocyclic rings, a fused ring of two or three heterocyclic rings. All or part of the fused ring can or can not have aromaticity. For example, a fused ring of three benzene rings can include the following structures:

[0059] The term "aromatic ring" means a carbocyclic ring having aromaticity. This includes, but is not limited to, benzene, naphthalene, anthracene, phenanthrene rings.

[0060] The term "heteroaromatic ring" means a heterocyclic ring having aromaticity.

[0061] The term "tricyclic fused aromatic heterocyclic ring" as used herein means a tricyclic fused heterocyclic ring having a fused aromatic group or a fused heteroaromatic group.

[0062] The term "alkyl", by itself or as part of another substituent, means, unless otherwise indicated, a saturated aliphatic hydrocarbon group having the number of carbon atoms designated (i.e., C 1-10 means a straight chain or branched chain (or combination thereof) of carbon atoms that is un-cyclized, saturated aliphatic hydrocarbon group of one to ten carbons. Examples of saturated hydrocarbon groups include, but are not limited to, the following groups, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, (cyclohexyl)methyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like homologs and isomers of the same. Preferred are alkyl groups containing 1 to 6 carbon atoms, further preferred are alkyl groups containing 1 to 3 carbon atoms. For example, C 1-3 alkyl means methyl, ethyl, n-propyl, and isopropyl.

[0063] Alkoxy is an alkyl group attached to the remainder of the molecule via an oxygen linker (-O-).

[0064] The term "benzo five-membered nitrogen heterocycle" refers to an organic compound structural unit that is fused from a benzene ring and a five-membered heterocycle containing a nitrogen atom. Examples include, but are not limited to, an indole ring, an isoindole ring, a benzopyrazole ring, a benzoxazole ring, a benzothiazole ring.

[0065] The term "cycloalkane" refers to a saturated or partially unsaturated monocyclic or polycyclic ring, and the term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, a cycloalkyl ring comprising 3 to 20 carbon atoms, preferably comprising 3 to 12 carbon atoms, more preferably comprising 3 to 8 (e.g., 3, 4, 5, 6, 7, and 8) carbon atoms, more preferably comprising 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spirocyclic, fused, and bridged cycloalkyl groups.

[0066] The term "heterocycloalkane" refers to a cycloalkane comprising at least one ring heteroatom (e.g., a nitrogen atom, an oxygen atom, or a sulfur atom). The term "heterocycloalkyl" refers to a cycloalkyl comprising at least one ring heteroatom (e.g., a nitrogen atom, an oxygen atom, or a sulfur atom). In some embodiments of the application, the heterocyclyl group is a 4-12 membered heterocyclyl group. In some embodiments of the application, the heterocyclyl group is a 4-6 membered heterocyclyl group, such as furanyl, imidazolidinyl, imidazolinyl, imidazolyl, isoquinolinyl, thiazolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, oxazolidinyl, oxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-oxadiazol-5(4H)-onyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, tetrahydrofuranyl, tetrazolyl, thienyl, tetrahydrothiophene, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, oxetanyl, oxetanyl, azetidinyl, azetidinyl, azetidinyl.

[0067] The term "halo" or "halogen", by itself or as part of another substituent, means, unless otherwise specified, a fluorine, chlorine, bromine, or iodine atom.

[0068] The compounds of the present application can exist in optically pure enantiomeric forms, pure diastereomeric forms, mixtures of enantiomers, mixtures of diastereomers, mixtures of enantiomeric racemates, racemates or mixtures of racemates. All possible isomers, stereoisomers, and mixtures thereof of the compounds of Formula (II) are within the scope of the present application. Metabolites of the compounds of Formula (II) and isotopologues of the compounds of Formula (II) are also within the scope of the present application.

[0069] The compounds of the present application can exist in unsolvated and solvated forms. The term'solvate' is used herein to describe a molecular complex comprising the compound of the present application and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol).

[0070] The compounds of the present application can exist in polymorphic forms, i.e., in more than one form or crystal structure.

[0071] The optically pure enantiomers, pure diastereomers of the present application can be obtained by chiral resolution or by chiral synthesis.

[0072] In some embodiments of the present application, the pure diastereomers of the compounds of the present application are obtained by chiral resolution, the resolution method is SFC resolution, the conditions are as follows:

[0073] For example, the compounds of Formula (II), stereoisomers, tautomers or pharmaceutically acceptable salts thereof, including compound 5, and the compounds 5-1 with retention time of 10.5 min and 5-2 with retention time of 17 min obtained after the resolution of compound 5 under the resolution conditions. In some embodiments of the present application, the compounds of the present application, stereoisomers, tautomers or pharmaceutically acceptable salts thereof can be isotopologues thereof (e.g., deuterated compounds).

[0074] In some embodiments of the present application, the compounds of the present application, stereoisomers, tautomers or pharmaceutically acceptable salts thereof can be metabolites thereof.

[0075] The compounds of the present application can be administered in crystalline or amorphous form. They can be given alone or in combination with one or more other compounds of the present application or in combination with one or more other drugs. Generally, they will be given as a formulation in association with one or more pharmaceutically acceptable excipients. The term 'excipient' is used herein to describe any ingredient other than the compound(s) of the application. The choice of excipient will to a large extent depend on the specific

[0076] The pharmaceutical composition according to the present application can be prepared in any form, for example, granules, powders, tablets, coated tablets, capsules, pills, syrups, drops, solutions, suspensions, and emulsions, or sustained-release preparations of the active ingredient.

[0077] The pharmaceutical composition according to the present application can further include one or more pharmaceutically acceptable carriers, which will be appropriately formulated to facilitate administration. The pharmaceutical composition according to the present application can also include pharmaceutically acceptable additives, such as diluents, lubricants, binders, glidants, disintegrants, sweeteners, flavoring agents, humectants, dispersants, surfactants, solvents, coating agents, foaming agents, or fragrances.

[0078] The pharmaceutical composition according to the present application can be administered by various routes according to conventional methods.

[0079] The term "isotopic derivative" refers to a derivative in which one or more atoms in a compound are replaced by an isotope (an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number that predominates in nature) of that atom. Examples of isotopes include, but are not limited to, isotopes of hydrogen (for example 2 H, 3 H), isotopes of carbon (for example 11 C, 13 C, and 14 C), isotopes of fluorine (for example 18 F), isotopes of nitrogen (for example 13 N, and 15 N), isotopes of oxygen (for example 15 O, 17 O, and 18 O).

[0080] The terms "pharmaceutically acceptable salt", "pharmaceutically acceptable salts" have the same meaning and refer to salts of active compounds that are produced in accordance with the particular substituents found on the compounds described herein, with relatively nontoxic acids or bases. When the compounds of the application contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or like salts. When the compounds of the application contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts are described in, e.g., Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19. Certain specific compounds of the application contain both basic and acidic functionalities, allowing the compounds to be converted into either base or acid addition salts.

[0081] "Pharmaceutically acceptable carrier and / or additive" refers to a material that aids in the administration of a compound to an individual and is absorbed by the individual, and can be included in a composition of the application without causing a significant adverse toxic effect to the patient.

[0082] JNJ-1802 as described herein refers to a compound with CAS number 2043343-94-6.

[0083] The compounds of the application exhibit good selectivity index (SI) in cellular potency evaluation, for example as shown in Figure 1. In addition, the compounds of the application exhibit better animal efficacy in inhibiting viral reoccurrence, for example as shown in Figure 2.

[0084] The term "treatment" means the successful treatment or amelioration of an injury, disease, pathology or condition, including any marker thereof, including any subjective or objective parameter, such as alleviation of symptoms; remission; decreased or slowing of rate of progression; lessening of extent of deterioration; improvement in a patient's physical or mental well-being. Treatment or amelioration of symptoms can be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric examination and / or psychiatric evaluation. The term "treatment" and its variants, can include prevention. In embodiments, treatment is prophylaxis. In embodiments, treatment does not include prophylaxis.

[0085] As used herein, and as is well understood in the art, "treatment" (also "treat") broadly includes any administration of a therapeutic agent to an individual for the purpose of effecting an advantageous or desired result, including clinical results. Advantageous or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, preventing spread or worsening of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total, and whether detectable or undetectable. In other words, "treatment" as used herein includes any cure, amelioration, or prevention of a disease. Treatment can prevent the disease from occurring; inhibit the disease's spread; relieve symptoms of the disease; completely or partially remove the causation, duration or extent of the disease; or a combination of these effects.

[0086] As used herein, "treatment" includes prophylactic treatment. Methods of treatment include administering to an individual a therapeutically effective amount of a compound described herein. The administration step can include a single administration or can include a series of administrations. The length of the treatment period depends on a variety of factors, such as the severity of the condition, the patient's age, the concentration of the compound, the activity of the composition used for treatment, and combinations thereof. It will also be appreciated that the effective amount of the agent for treatment or prevention can increase or decrease over the course of the particular treatment or prevention protocol. Changes in dosage can be observed by standard diagnostic assays known in the art and become evident by the effect on the patient.

[0087] The term "prevention" refers to reducing the occurrence of symptoms of a disease in a patient. As described above, prevention can be complete (no detectable symptoms) or partial, such that fewer symptoms are observed than would occur without treatment. In embodiments, prevention refers to slowing the progression of a disease, disorder, or condition, or inhibiting its progression to a deleterious or other undesirable state.

[0088] A "patient" or "individual in need thereof refers to a living organism suffering from or susceptible to a disease or condition that can be treated by administration of a pharmaceutical composition provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammalian animals. In some embodiments, the patient is a human.

[0089] One of ordinary skill in the art of infectious disease treatment will be able to determine an effective amount from the test results presented below. In general, it is expected that the effective amount will be in the range of 0.01 mg / kg to 50 mg / kg body weight, more preferably 0.1 mg / kg to 10 mg / kg body weight, per day. The required dose can be given in appropriate division as two, three, four or more sub-doses at appropriate intervals throughout the day. The sub-doses can be formulated as unit dosage forms, for example, containing 1 to 1000 mg, and particularly 5 mg to 200 mg, of active ingredient in each unit dose.

[0090] The precise dosage and frequency of administration depends on the particular compound of formula (II) employed, the particular condition being treated, the severity of the condition being treated, the age, the body weight, and the general physical condition of the particular patient, as well as other medication that the individual can be taking, as is well known to those skilled in the art. Additionally, it is apparent to those skilled in the art that the

[0091] The present application is further exemplified by the following examples, which are illustrative, and in no way limit the scope or use of the present application. Any modification or change, which is readily apparent to one of ordinary skill in the art, made to the present application is intended to fall within the scope of this application.

[0092] The starting materials or reagents used in the examples of the present application are commercially available unless otherwise stated.

[0093] The abbreviations used in the present application have the conventional meaning in the art, for example the following abbreviations have the following meanings:

[0094] Process for the preparation of compounds

[0095] Example 1: Synthesis of compound 1

[0096] Synthesis of 4-chloro-2-methoxyphenylacetyl chloride 1b

[0097] 4-chloro-2-methoxyphenylacetic acid 1a (1.0 g, 5.0 mmol) was added to sulfurous chloride (10 mL) and stirred at 60 °C for 0.5 h. After monitoring the reaction completion by LC-MS, the compound 1b (1.1 g) was obtained by concentration.

[0098] Synthesis of 2-iodo-4-(trifluoromethoxy)aniline 1d

[0099] To a solution of 4-trifluoromethoxyaniline 1c (10 g, 56 mmol) in ethanol (100 mL) was added silver sulfide (18 g, 56 mmol), I2(14 g, 56 mmol) and stirred at room temperature under nitrogen overnight. After monitoring the reaction completion by LC-MS, the reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (100 mL*3). The mixture was concentrated under reduced pressure and purified by silica gel chromatography (PE:EA = 5:1 elution) to give compound 1d (8.6 g).

[0100] MS (ESI) m / z: 304.0 [M+H] + .

[0101] 2,2,2-trifluoro-N-(2-iodo-4-trifluoromethoxyphenyl)acetamide If

[0102] To a solution of 2-iodo-4-(trifluoromethoxy)aniline Id (8.6 g, 28 mmol) in dichloromethane (100 mL), triethylamine (7.2 g, 71 mmol) and trifluoroacetic anhydride Ie (8.9 g, 43 mmol) were added and stirred at room temperature for 0.5 h. The reaction was monitored by LC-MS and completed. The reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (100 mL*3). The mixture was concentrated under reduced pressure and purified by silica gel chromatography (PE:EA = 5:1 elution) to give compound If (10 g).

[0103] MS (ESI) m / z: 400.0 [M+H] + .

[0104] Synthesis of 3-(5-(trifluoromethoxy)-lH-indol-2-yl)-l-propanol Ih

[0105] To a solution of 2,2,2-trifluoro-N-(2-iodo-4-trifluoromethoxyphenyl)acetamide If (1.0 g, 2.5 mmol) in triethylamine (20 mL), n-pent-4-yn-l-ol Ig (250 mg, 3.0 mmol), [l,l'-bis(diphenylphosphino)ferrocene]palladium dichloride (180 mg, 0.25 mmol), and cuprous iodide (950 mg, 5.0 mmol) were added and stirred at 80 °C under nitrogen overnight. The reaction was monitored by LC-MS and completed. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL*3). The mixture was concentrated under reduced pressure and purified by reverse phase column chromatography to give compound Ih (270 mg).

[0106] MS (ESI) m / z: 400.0 [M+H] + .

[0107] Synthesis of 3-(5-(trifluoromethoxy)-lH-indol-2-yl)propyl methanesulfonate Ii

[0108] To a solution of 3-(5-(trifluoromethoxy)-lH-indol-2-yl)-l-propanol Ih (270 mg, 1.0 mmol) in tetrahydrofuran (6.0 mL), triethylamine (160 mg, 1.6 mmol), and methanesulfonyl chloride (140 mg, 1.2 mmol) were added and stirred at 0 °C for 0.5 h. The reaction was monitored by LC-MS and completed. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL*3). Concentration and drying gave compound Ii (350 mg).

[0109] MS (ESI) m / z: 338.0 [M+H] + .

[0110] Synthesis of 7-(trifluoromethoxy)-2,3-dihydro-1H-pyrrolo[1,2-a]indol-1-ium 1j

[0111] To a solution of 3-(5-(trifluoromethoxy)-1H-indol-2-yl)propyl methanesulfonate 1i (350 mg, 1.0 mmol) in tetrahydrofuran (10 mL) was added potassium tert-butoxide (230 mg, 2.1 mmol) and stirred at room temperature for 0.5 h. After the reaction was completed by LC-MS, the reaction mixture was diluted with water (60 mL) and extracted with ethyl acetate (30 mL*3). The mixture was concentrated under reduced pressure and purified by preparative thin layer chromatography to give compound 1j (240 mg).

[0112] MS (ESI) m / z: 338.0 [M+H] + .

[0113] 1 H NMR (400 MHz, CDCl3) δ 7.38 (s, 1H), 7.16 (d, J = 8.7 Hz, 1H), 6.97 (dd, J = 8.7, 1.2 Hz, 1H), 6.16 (d, J = 0.7 Hz, 1H), 4.09-3.99 (m, 2H), 3.01 (dd, J = 11.1, 4.1 Hz, 2H), 2.60 (ddd, J = 11.4, 8.9, 5.4 Hz, 2H).

[0114] Synthesis of 2-(4-chloro-2-methoxyphenyl)-1-(7-(trifluoromethoxy)-2,3-dihydro-1H- pyrrolo[1,2-a]indol-9-yl)ethanone 1k

[0115] To a solution of 7-(trifluoromethoxy)-2,3-dihydro-1H-pyrrolo[1,2-a]indol-1-ium 1j (240 mg, 1.0 mmol) in dichloromethane (10 mL) was added diethylaluminum chloride (1.5 mL, 1.5 mmol, 1.0 M) at 0 °C and stirred at 0 °C for 0.5 h under nitrogen protection. Then 4-chloro-2-methoxyphenylacetyl chloride 1b (330 mg, 1.5 mmol) was added and stirred at room temperature for 1 h. After the reaction was completed by LC-MS, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL*3). The mixture was concentrated under reduced pressure and purified by reverse phase column chromatography to give compound 1k (190 mg).

[0116] MS (ESI) m / z: 338.0 [M+H] + .

[0117] Synthesis of 2-bromo-2-(4-chloro-2-methoxyphenyl)-1-(7-(trifluoromethoxy)- 2,3-dihydro-1H-pyrrolo[1,2-a]indol-9-yl)ethanone 1l

[0118] In a solution of 2-(4-chloro-2-methoxyphenyl)-1-(7-(trifluoromethoxy)-2,3- dihydro-1H-pyrrolo[1,2-a]indol-9-yl)ethanone 1k (190 mg, 0.45 mmol) in 1,4- dioxane (20 mL), copper bromide (400 mg, 1.8 mmol) was added, and stirred at 80 °C for 4 h under nitrogen protection. After the reaction was completed by LC-MS, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL*3). The mixture was concentrated under reduced pressure and purified by reverse phase column chromatography to give compound 1l (180 mg).

[0119] MS (ESI) m / z: 502.0 [M+H] + .

[0120] Synthesis of compound 1

[0121] In a solution of 2-bromo-2-(4-chloro-2-methoxyphenyl)-1-(7-(trifluoromethoxy)- 2,3-dihydro-1H-pyrrolo[1,2-a]indol-9-yl)ethanone 1l (180 mg, 0.37 mmol) in acetonitrile (20 mL), 3-methoxy-5-(methylsulfonyl)aniline 1m (150 mg, 0.73 mmol), N,N- diisopropylethylamine (140 mg, 1.1 mmol) were added, and stirred at 90 °C overnight under nitrogen protection. After the reaction was completed by LC-MS, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL*3). The mixture was concentrated under reduced pressure and purified by reverse phase column chromatography to give compound 1 (12 mg).

[0122] MS (ESI) m / z: 623.4 [M+H] + .

[0123] HPLC: 254 nm: 97.805%, 214 nm: 97.444%.

[0124] 1H NMR (400 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.55 (d, J = 8.8 Hz, 1H), 7.27 (d, J = 8.2 Hz, 1H), 7.21 (dd, J = 8.7, 1.8 Hz, 1H), 7.13 (dd, J = 5.1, 3.1 Hz, 2H), 7.00 (dd, J = 8.2, 2.0 Hz, 1H), 6.86 (s, 1H), 6.65 - 6.59 (m, 1H), 6.38 (s, 1H), 6.04 (d, J = 8.3 Hz, 1H), 4.20 (t, J = 7.3 Hz, 2H), 3.86 (s, 3H), 3.71 (s, 3H), 3.30 - 3.23 (m, 2H), 3.10 (s, 3H), 2.63 (dt, J = 14.1, 7.0 Hz, 2H).

[0125] Example 2: Synthesis of compound 2

[0126] Synthesis of 2-(2-nitronaphthalen-l-yl)acetonitrile 2b

[0127] A solution of potassium tert-butoxide (2.6 g, 23 mmol) in dry N,N- dimethylformamide (30 mL) was cooled to -20 °C, then a solution of 2-nitronaphthalene 2a (2.0 g, 12 mmol) and chloroacetonitrile (1.1 g, 14 mmol) in dry N,N-dimethylformamide (10 mL) was added dropwise over 2-3 min and stirred at -20 °C to -10 °C for 5-6 min. After the reaction was completed by LC-MS, the mixture was poured into 1.0 N hydrochloric acid (300 mL) and extracted with ethyl acetate (300 mL). The organic layer was washed with brine (300 mL*3), dried over sodium sulfate and concentrated under reduced pressure, then purified by flash column chromatography (PE:DCM = 100:0 to 30:70) to give compound 2b (1.2 g).

[0128] 1 H NMR (400 MHz, CDCl3) δ 8.23 (d, J = 8.5 Hz, 1H), 8.06-7.94 (m, 3H), 7.84-7.74 (m, 2H), 4.41 (s, 2H).

[0129] Synthesis of 3H-benzo[E]indole 2c

[0130] To a mixture of 2-(2-nitronaphthalen-l-yl)acetonitrile 2b (1.2 g, 5.8 mmol), ethanol (30 mL) and water (3.0 mL) was added Pd / C (1.0 g) and acetic acid (0.50 mL) and purged with H2for 3 min. The mixture was stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was diluted with 0.10 M potassium hydroxide solution (40 mL) and extracted with dichloromethane (100 mL). The organic layer was concentrated under reduced pressure. The residue was purified by flash column chromatography (PE:EA = 100:0 to 60:40) to give compound 2c (630 mg).

[0131] MS (ESI) m / z: 168.0 [M+H] + .

[0132] 1 H NMR (400 MHz, CDC13) δ 8.47 (s, 1H), 8.24 (d, J = 8.2 Hz, 1H), 7.90 (d, J = 8.1 Hz, 1H), 7.63-7.48 (m, 3H), 7.43-7.39 (m, 1H), 7.27 (t, J = 2.8 Hz, 1H), 7.10-7.09 (m, 1H).

[0133] Synthesis of l-(3H-benzo[E]indol-l-yl)-2-(4-chloro-2-methoxyphenyl)ethanone 2e

[0134] To a solution of 3H-benzo[E]indole 2c (450 mg, 2.4 mmol) in dry dichloromethane (15 mL) was added diethylaluminum chloride (3.6 mL, 3.6 mmol, 1.0 M) dropwise at 0 °C and the mixture was stirred at 0 °C for 30 min. A solution of 4-chloro-2-methoxyphenylacetyl chloride 2d (790 mg, 3.6 mmol) in dry dichloromethane (4.0 mL) was added dropwise at 0 °C and stirred at 0 °C for 1 h. After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL*2). The organic layer was concentrated under reduced pressure. The residue was purified by flash column chromatography (PE:EA = 100:0 to 30:70) to give compound 2e (540 mg).

[0135] MS (ESI) m / z: 350.0 [M+H] + .

[0136] 1H NMR (400 MHz, DMSO-d6) δ 12.42 (s, 1H), 9.63 (d, J = 8.0 Hz, 1H), 8.60 (d, J = 3.2 Hz, 1H), 7.93 (dd, J = 8.1, 1.1 Hz, 1H), 7.69 (dd, J = 23.1, 8.8 Hz, 2H), 7.51 - 7.38 (m, 2H), 7.27 (d, J = 8.0 Hz, 1H), 7.08 (d, J = 2.0 Hz, 1H), 7.00 (dd, J = 8.0, 2.0 Hz, 1H), 4.34 (s, 2H), 3.77 (s, 3H).

[0137] Synthesis of 1 -(3H-benzo[E]indol-1 -yl)-2-bromo-2-(4-chloro-2- methoxyphenyl)ethanone 2f

[0138] To a solution of 1 -(3H-benzo[E]indol-1 -yl)-2-(4-chloro-2- methoxyphenyl)ethanone 2e (100 mg, 0.28 mmol) in 1,4-dioxane (20 mL) was added CuBr2(130 mg, 0.57 mmol) and the mixture was stirred at 80 °C for 3 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (PE:DCM = 100:0 to 28:72) to afford compound 2f (40 mg).

[0139] MS (ESI) m / z: 428.0; 430.0 [M+H; M+H+1] + .

[0140] Synthesis of compound 2

[0141] To a solution of 1 -(3H-benzo[E]indol-1 -yl)-2-bromo-2-(4-chloro-2- methoxyphenyl)ethanone 2f (40 mg, 0.093 mmol) in acetonitrile (10 mL) was added compound 1 m (38 mg, 0.18 mmol) and diisopropylethylamine (36 mg, 0.28 mmol). The mixture was stirred at 90 °C for 22 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and the residue was purified by preparative thin layer chromatography (DCM) and reverse phase column to afford compound 2 (10 mg).

[0142] MS (ESI) m / z: 549.3 [M+H] + .

[0143] HPLC: 214 nm: 99.391 %, 254 nm: 99.351 %.

[0144] 1H NMR (400 MHz, DMSO-d6) δ 12.50 (s, 1H), 9.55 (d, J = 8.4 Hz, 1H), 8.64 (s, 1H), 7.93 (d, J = 8.1 Hz, 1H), 7.72 (d, J = 8.8 Hz, 1H), 7.63 (d, J = 8.7 Hz, 1H), 7.53 (dd, J = 11.2, 4.2 Hz, 1H), 7.49 - 7.39 (m, 2H), 7.11 (dd, J = 12.1, 4.9 Hz, 2H), 7.01 - 6.92 (m, 2H), 6.66 (s, 1H), 6.60 (d, J = 1.6 Hz, 1H), 6.44 (d, J = 7.8 Hz, 1H), 3.98 (s, 3H), 3.74 (s, 3H), 3.10 (s, 3H).

[0145] Example 3: Synthesis of compound 3

[0146] Synthesis of 4-(5-(trifluoromethoxy)-1H-indol-2-yl)-1-butanol 3b

[0147] To a solution of 2,2,2-trifluoro-N-(2-iodo-4-trifluoromethoxyphenyl)acetamide 1f (3.0 g, 7.5 mmol) in triethylamine (40 mL) was added n-hex-5-yn-1-ol 3a (890 mg, 9.0 mmol), [1,1'- bis(diphenylphosphino)ferrocene]palladium dichloride (550 mg, 0.75 mmol) and cuprous iodide (2.8 g, 15 mmol), stirred at 80 °C overnight under nitrogen protection. After the reaction was detected to be completed by LC-MS, the reaction mixture was diluted with water (200 mL) and extracted with EA (50 mL*3). The mixture was concentrated under reduced pressure and purified by a reverse phase column to give compound 3b (1.4 g).

[0148] MS (ESI) m / z: 274.0 [M+H] + .

[0149] Synthesis of 4-(5-(trifluoromethoxy)-1H-indol-2-yl)butyl methanesulfonate 3c

[0150] To a solution of 4-(5-(trifluoromethoxy)-1H-indol-2-yl)-1-butanol 3b (1.4 g, 5.1 mmol) in dichloromethane (15 mL) was added diisopropylethylamine (1.3 g, 10 mmol) and methanesulfonyl chloride (880 mg, 7.7 mmol) at 0 °C, stirred at 0 °C for 0.5 h. After the reaction was detected to be completed by thin layer chromatography (PE:EA = 5:1), the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (50 mL*3). Concentration and drying gave compound 3c (1.8 g).

[0151] 2-(trifluoromethoxy)-6,7,8,9-tetrahydropyrido[l,2-a]indole 3d

[0152] To a solution of 4-(5-(trifluoromethoxy)-lH-indol-2-yl)butyl methanesulfonate 3c (1.8 g, 5.1 mmol) in tetrahydrofuran (20 mL) was added potassium tert-butoxide (1.1 g, 10 mmol) and stirred at room temperature for 0.5 h. After the reaction was detected by LC-MS, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (50 mL*3). The mixture was concentrated under reduced pressure and purified by silica gel column (PE:EA = 4: 1) to give compound 3d (940 mg).

[0153] MS (ESI) m / z: 256.0 [M+H] + .

[0154] Synthesis of 2-(4-chloro-2-methoxyphenyl)-l-(2-(trifluoromethoxy)-6,7,8,9- tetrahydropyrido[l,2-a]indol-10-yl)ethanone 3e

[0155] To a solution of 2-(trifluoromethoxy)-6,7,8,9-tetrahydropyrido[l,2-a]indole 3d (400 mg, 1.6 mmol) in dichloromethane (8.0 mL) was added diethylaluminum chloride (2.3 mL, 2.3 mmol, 1.0 M) under nitrogen protection and stirred at 0 °C for 0.5 h. Then 4-chloro-2-methoxyphenylacetyl chloride lb (510 mg, 2.3 mmol) was added and stirred at room temperature for 2 h. After the reaction was detected by LC-MS, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (50 mL*3). The mixture was concentrated under reduced pressure and purified by reverse phase column to give compound 3e (560 mg).

[0156] MS (ESI) m / z: 438.2 [M+H] + .

[0157] Synthesis of 2-bromo-2-(4-chloro-2-methoxyphenyl)-l-(2-(trifluoromethoxy)-6,7,8,9- tetrahydropyrido[l,2-a]indol-10-yl)ethanone 3f

[0158] In a solution of 2-(4-chloro-2-methoxyphenyl)-l-(2-(trifluoromethoxy)-6,7,8,9- tetrahydropyrido[l,2-a]indol-l 0-yl)ethanone 3e (560 mg, 1.3 mmol) in 1,4-dioxane (20 mL), copper bromide (850 mg, 3.8 mmol) was added, and the mixture was stirred at 80 °C under nitrogen protection for 2 h. After the reaction was completed by LC-MS, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL*3). The mixture was concentrated under reduced pressure and purified by a reverse phase column to give compound 3f (350 mg).

[0159] MS (ESI) m / z: 516.0 [M+H] + .

[0160] Synthesis of compound 3

[0161] In a solution of 2-bromo-2-(4-chloro-2-methoxyphenyl)-l-(2-(trifluoromethoxy)-6,7,8,9- tetrahydropyrido[l,2-a]indol-l 0-yl)ethanone 3f (350 mg, 0.69 mmol) in acetonitrile (40 mL), 3-methoxy-5-(methylsulfonyl)aniline Im (280 mg, 1.4 mmol) and diisopropylethylamine (270 mg, 2.1 mmol) were added, and the mixture was stirred at 80 °C under nitrogen protection overnight. After the reaction was completed by LC-MS, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL*3). The mixture was concentrated under reduced pressure and purified by a reverse phase column to give compound 3 (15 mg).

[0162] MS (ESI) m / z: 637.0 [M+H] + .

[0163] HPLC: 254 nm: 96.094%, 214 nm: 96.338%.

[0164] 1H NMR (400 MHz, DMSO) δ 7.83 (s, 1H), 7.60 (d, J = 8.9 Hz, 1H), 7.23 - 7.17 (m, 2H), 7.13 (d, J = 2.0 Hz, 1H), 7.09 (d, J = 8.4 Hz, 1H), 6.98 (dd, J = 8.2, 2.0 Hz, 1H), 6.84 (s, 1H), 6.65 - 6.60 (m, 1H), 6.39 (s, 1H), 6.10 (d, J = 8.3 Hz, 1H), 4.25 - 4.06 (m, 2H), 3.85 (s, 3H), 3.70 (s, 3H), 3.25 (t, J = 6.2 Hz, 2H), 3.09 (s, 3H), 2.03 (d, J = 5.7 Hz, 2H), 1.93 - 1.80 (m, 2H).

[0165] Example 4: Synthesis of compound 4

[0166] Synthesis of compound 4 was according to reference example 1.

[0167] MS (ESI) m / z: 623.1 [M+H] + .

[0168] Example 5: Synthesis of compound 5

[0169] Synthesis of N,N-dimethyl-2-(1-nitronaphthalen-2-yl)vinyl-1-amine 5b

[0170] To a mixture of 2-methyl-1-nitronaphthalene 5a (8.0 g, 43 mmol) in N,N- dimethylformamide (30 mL) was added DMF-DMA (16 mL). The mixture was stirred at 140 °C for 41 h. After TLC detection of the reaction completion, the reaction mixture was diluted with water (300 mL) and extracted with ethyl acetate (300 mL). The organic layer was washed with brine (300 mL*3). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give compound 5b (9.5 g).

[0171] Synthesis of 1H-benzo[G]indole 5c

[0172] To a mixture of N,N-dimethyl-2-(l-nitronaphthalen-2-yl)vinyl-l-amine 5b (9.5 g, 39 mmol) in ethanol (150 mL) and water (30 mL) was added zinc powder (13 g, 200 mmol) and ammonium chloride (21 g, 390 mmol). The mixture was stirred at 80 °C for 2 h. After the reaction was completed, the mixture was filtered. The filtrate was diluted with water (300 mL) and extracted with ethyl acetate (300 mL). The organic layer was concentrated under reduced pressure, and the residue was purified by silica gel column (PE:EA = 50:1 ~ 20:1) to give compound 5c (2.0 g).

[0173] MS (ESI) m / z: 168.2 [M+H] + .

[0174] 1 H NMR (400 MHz, DMSO-d6) δ 12.01 (s, 1H), 8.36 (d, J = 8.2 Hz, 1H), 7.91 (d, J = 8.1 Hz, 1H), 7.67 (d, J = 8.6 Hz, 1H), 7.52 (ddd, J = 8.2, 7.0, 1.2 Hz, 1H), 7.47 - 7.36 (m, 3H), 6.59 (dd, J = 2.9, 2.0 Hz, 1H).

[0175] Synthesis of l-(lH-benzo[G]indol-3-yl)-2-(4-chloro-2-methoxyphenyl)ethanone 5d

[0176] To a solution of lH-benzo[G]indole 5c (1.0 g, 6.0 mmol) in dry dichloromethane (40 mL) was added diethylaluminum chloride (9.0 mL, 9.0 mmol, 1.0 M) dropwise at 0 °C and stirred for 30 min at 0 °C. Then a solution of 4-chloro-2-methoxyphenylacetyl chloride lb (2.0 g, 9.0 mmol) in dry dichloromethane (10 mL) was added dropwise at 0 °C and stirred for 1 h. After the reaction was completed, the reaction mixture was diluted with water (300 mL) and extracted with ethyl acetate (300 mL*2). The organic phase was concentrated under reduced pressure, and the residue was purified by reverse phase column to give compound 5d (850 mg).

[0177] MS (ESI) m / z: 350.0 [M+H] + .

[0178] 1H NMR (400 MHz, DMSO-d6) δ 12.81 (s, 1H), 8.50 (d, J = 2.9 Hz, 1H), 8.42 (d, J = 8.2 Hz, 1H), 8.25 (d, J = 8.7 Hz, 1H), 7.97 (d, J = 8.1 Hz, 1H), 7.62 (dd, J = 15.1, 8.0 Hz, 2H), 7.49 (t, J = 7.5 Hz, 1H), 7.24 (d, J = 8.0 Hz, 1H), 7.07 (d, J = 1.9 Hz, 1H), 6.98 (dd, J = 8.0, 1.9 Hz, 1H), 4.25 (s, 2H), 3.77 (s, 3H).

[0179] Synthesis of 1-(1H-benzo[G]indol-3-yl)-2-bromo-2-(4-chloro-2- methoxyphenyl)ethanone 5e

[0180] To a solution of 1-(1H-benzo[G]indol-3-yl)-2-(4-chloro-2- methoxyphenyl)ethanone 5d (850 mg, 2.4 mmol) in 1,4-dioxane (50 mL) was added copper bromide (1.0 g, 49 mmol). The mixture was stirred at 80 °C for 2 h. After the reaction was completed, the reaction mixture was diluted with water (150 mL) and extracted with ethyl acetate (150 mL*2). The organic phase was concentrated under reduced pressure, and the residue was purified by reverse phase column (FA) to give compound 5e (320 mg).

[0181] MS (ESI) m / z: 428.0; 430.0 [M+H; M+2+H] + .

[0182] 1 H NMR (400 MHz, DMSO-d6) δ 12.98 (s, 1H), 8.40 (dd, J = 13.6, 5.7 Hz, 2H), 8.26 (d, J = 8.7 Hz, 1H), 7.99 (d, J = 7.9 Hz, 1H), 7.69 (d, J = 8.7 Hz, 1H), 7.61 (t, J = 7.5 Hz, 2H), 7.50 (t, J = 7.5 Hz, 1H), 7.15 (d, J = 2.0 Hz, 1H), 7.08 (dd, J = 8.4, 2.0 Hz, 1H), 6.96 (s, 1H), 3.91 (s, 3H).

[0183] Synthesis of compound 5

[0184] To a solution of 1-(1H-benzo[G]indol-3-yl)-2-bromo-2-(4-chloro-2- methoxyphenyl)ethanone 5e (270 mg, 0.63 mmol) in acetonitrile (80 mL) was added 3-methoxy-5-(methylsulfonyl)aniline 1m (250 mg, 1.3 mmol) and diisopropylethylamine (160 mg, 1.3 mmol). The mixture was stirred at 90 °C overnight. After completion of the reaction, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL). The organic phase was concentrated under reduced pressure and the residue was purified by reverse phase column chromatography and preparative thin layer chromatography (dichloromethane) to obtain compound 5 (55 mg).

[0185] MS (ESI) m / z: 549.0 [M+H] + .

[0186] HPLC: 214 nm: 99.109%, 254 nm: 99.356%.

[0187] 1 H NMR (400 MHz, DMSO-d6) δ 12.96 (s, 1H), 8.50 (s, 1H), 8.39 (d, J = 8.2 Hz, 1H), 8.28 (d, J = 8.7 Hz, 1H), 7.97 (d, J = 8.1 Hz, 1H), 7.67 (d, J = 8.8 Hz, 1H), 7.60 (t, J = 7.5 Hz, 1H), 7.49 (t, J = 7.5 Hz, 1H), 7.39 (d, J = 8.3 Hz, 1H), 7.13 (dd, J = 13.3, 4.6 Hz, 2H), 6.97 (dd, J = 8.9, 7.3 Hz, 2H), 6.62 (d, J = 15.8 Hz, 2H), 6.34 (d, J = 7.7 Hz, 1H), 4.04 (s, 3H), 3.73 (s, 3H), 3.10 (s, 3H).

[0188] Synthesis of compounds 5-1 and 5-2

[0189] Compound 5 (55 mg) was resolved by SFC to give compound 5-1 (18.9 mg) with retention time 10.5 min and compound 5-2 (18.9 mg) with retention time 17 min. The resolution method is as follows:

[0190] Compound 5-1:

[0191] LCMS (ESI+): m / z = 549.2 [M+H] +

[0192] 1H NMR (400 MHz, DMSO-d6) δ 12.95 (s, 1H), 8.49 (d, J = 3.2 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 8.28 (d, J = 8.8 Hz, 1H), 7.97 (d, J = 8.4 Hz, 1H), 7.67 (d, J = 8.8 Hz, 1H), 7.60 (t, J = 6.8 Hz, 1H), 7.49 (t, J = 6.8 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.14 (d, J = 2.0 Hz, 1H), 7.11 (d, J = 8.0 Hz, 1H), 6.98 (dd, J = 8.4, 2.0 Hz, 1H), 6.95 (s, 1H), 6.64 (s, 1H), 6.60 (s, 1H), 6.34 (d, J = 8.0 Hz, 1H), 4.04 (s, 3H), 3.73 (s, 3H), 3.10 (s, 3H).

[0193] Compound 5-2:

[0194] LCMS (ESI+): m / z = 549.2 [M+H] +

[0195] 1 H NMR (400 MHz, DMSO-d6) δ 12.95 (s, 1H), 8.49 (d, J = 3.2 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 8.28 (d, J = 8.8 Hz, 1H), 7.97 (d, J = 8.4 Hz, 1H), 7.67 (d, J = 8.8 Hz, 1H), 7.60 (t, J = 6.8 Hz, 1H), 7.49 (t, J = 6.8 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.14 (d, J = 2.0 Hz, 1H), 7.11 (d, J = 8.0 Hz, 1H), 6.98 (dd, J = 8.4, 2.0 Hz, 1H), 6.95 (s, 1H), 6.64 (s, 1H), 6.60 (s, 1H), 6.34 (d, J = 8.0 Hz, 1H), 4.04 (s, 3H), 3.73 (s, 3H), 3.10 (s, 3H).

[0196] Example 6: Synthesis of compound 6

[0197] Synthesis of compound 6 is described in reference example 5.

[0198] MS (ESI) m / z: 553.1 [M+H] + .

[0199] Example 7: Synthesis of compound 7

[0200] Synthesis of compound 7 Reference Example 5.

[0201] MS (ESI) m / z: 551.1 [M+H] + .

[0202] Example 8: Synthesis of compound 8

[0203] Synthesis of compound 8 Reference Example 5.

[0204] MS (ESI) m / z: 549.1 [M+H] + .

[0205] Biological Example

[0206] Experiment 1: Dengue virus amplification and titer determination

[0207] Dengue viruses used in the experiment were DENV-2 New Guinea, DENV-3 H87 strain, DENV-1 Hawaii strain and DENV4 H241 strain. Dengue viruses were amplified in C6 / 36 cells and the virus titer was determined by immunoplaque assay on Vero cells. The steps of immunoplaque assay virus titer determination were as follows: Vero cells were plated in 48-well plates one day in advance, 5x10 4 cells / well, 200 μL / well, in 10% FBS DMEM medium, 37°C, 5% CO2 culture, and the cell confluence was 80% when the experiment was performed. The virus was gradiently diluted with 2% FBS DMEM medium, 100 μL / well of the virus dilution was added, and incubation was performed for 2-4 h. The supernatant was aspirated, 500 μL / well of 2% carboxymethyl cellulose was added, 37°C, 5% CO2 culture was performed for 5-6 days. The carboxymethyl cellulose was aspirated, PBS was used for washing, and 4% paraformaldehyde was used for fixation for 30 min. The supernatant was aspirated, PBS was used for washing for 3 times, 0.3% Triton-X100 and 5% BSA were added for permeation blocking, and incubation was performed at room temperature for 1 h. The supernatant was aspirated, the secondary antibody dilution (AS003) was added, incubation was performed at room temperature on a shaker for 2 h, and light shielding was performed. The supernatant was aspirated, PBS was used for washing for 3 times, 80-100 μL / well of KPL True Blue was added, color development was performed at room temperature on a shaker for 15 min. The supernatant was aspirated, tap water was used for washing for 1 time, color development was terminated, the number of plaques was counted, and the virus titer was calculated.

[0208] Experiment 2: Anti-dengue virus cell efficacy evaluation

[0209] Vero cells were cultured in DMEM medium with 5% CO2 at 37℃ and 5% FBS. Cell counting was performed using Bio-Rad C100-SE / C100, and 200 μL of cell suspension was inoculated into each well of a 48-well plate at a cell density of 2 x 10 5 cells / mL. Subsequent experiments were performed when the cells reached 80% confluence. The compounds were gradiently diluted with DMEM medium containing 2% FBS, and 6-8 concentration gradients were set. The medium was aspirated, 120 μL / well of the compound was added for incubation for 1 h, and blank control wells and virus control wells (DMEM medium containing 2% FBS was added) were additionally set. The dengue virus was diluted with DMEM medium containing 2% FBS to 5 x 10 5 PFU / mL according to the virus titer determined in Experiment 1, and 40 μL / well of the diluted virus was inoculated, i.e., MOI = 0.3, for infection at 37℃ for 2 h. Two duplicate wells were set for each drug concentration, and the culture was continued for 48 h. The supernatant was collected at 48 hpi and added to the lysis well of a Vazyme Virus DNA / RNA Extraction Kit 2.0 (Prepackaged) RM501 reagent box, and the RM501 program was run. After the program ended, the RNA was aspirated from the elution well and directly subjected to QRT-PCR detection or stored in a refrigerator at -20℃. According to the instructions, QRT-PCR detection was performed using a Vazyme Hiscript IIOne Step qRT-PCR SYBR Green Kit reagent box.

[0210] The QRT-PCR results were processed using Design and Analysis software, a standard curve was drawn, and the virus copy number of each well was exported to an Excel table. In Excel, the inhibition rate of the compound on the virus in each well was calculated using the formula: inhibition rate = 1 - virus copy number of the compound well / virus copy number of the virus well. The "dose-inhibition rate" curve was constructed using GraphPad Prism software: the concentration of the compound was taken as the logarithm of the X axis, and the corresponding virus inhibition rate was taken as the Y axis. The S curve was fitted, and the half maximal effective concentration EC 50 value of the compound was calculated, as shown in Table 1. In combination with the half maximal cytotoxicity concentration CC 50 value obtained by the cell viability test, the compound selection index SI was calculated, SI = CC 50 / EC 50 , and a typical curve graph of the compound is shown in FIG. 1.

[0211] Table 1: Cell efficacy analysis of the compounds of the present application and JNJ-1802 against different serotypes of dengue virus

[0212] Experiment Three: Compound Cytotoxicity Test

[0213] Cells were plated one day in advance in 96-well plates at 2 x 10 5 cells / mL. Cells were incubated at 37°C, 5% CO2for 16 hours, and the confluency was 70% before the experiment. The compounds were gradiently diluted with 2% FBS DMEM medium, 2-fold dilution, and 6-8 concentration gradients were set. The medium was aspirated, and the gradiently diluted compounds were added to the experimental groups, and 2% FBS DMEM medium was added to the cell control group, 100 μL / well, 37°C, 5% CO2incubation for 48 hours. The CCK-8 solution was 10-fold diluted with 2% FBS DMEM medium. The supernatant in the 96-well plate was aspirated, and 100 μL / well of the diluted CCK-8 working solution was added, 37°C, 5% CO2incubation for 1.5 hours. The absorbance at 450 nm was detected using a microplate reader, and the data was exported. According to the readings of the microplate reader, the cell activity was calculated: cell activity = (experimental group - blank group) / (control group - blank group). The "dose-cell activity" curve was constructed using GraphPad Prism software: the concentration of the compound was taken as the X-axis, and the corresponding cell activity was taken as the Y-axis, and the S curve was fitted to calculate the half-maximal cytotoxicity concentration CC 50 .

[0214] Table 2: Toxicity test of the compounds of the present application and JNJ-1802 in multiple cells

[0215] Experiment Four: In vivo antiviral efficacy evaluation of compounds in DENV-2 infected AG129 mouse model

[0216] The mouse model of dengue virus infection used AG129 mice, male, 8-10 weeks old. The animal experiment was carried out in an ABSL-2 level animal experiment facility, and the mice were adapted for three days before the experiment, and the state was observed every day, and the weight was measured. The compound was dissolved in DMSO to prepare a stock solution of 10 mg / mL, and stored at -20°C for standby. The solvent was prepared with 80% PEG400 and 20% sterilized water, and before each administration, the stock solution was diluted with the solvent to 0.2 mg / mL, and orally administered by gavage, twice a day, and the daily administration dose was 4 mg / kg. One hour after administration on day 0, each mouse was injected intraperitoneally with DENV-2 at a viral dose of 2 x 10 7PFU, twice daily thereafter until the end of dosing on day 5. The body weight of mice was weighed daily, and the health signs of mice were evaluated. After the end of dosing on day 3, 200 μL of blood was taken from the orbit of each mouse, centrifuged at 3000 rpm for 10 min to obtain plasma, and the viral nucleic acid in the plasma was extracted and the viral load was detected by QRT-PCR to evaluate the antiviral efficacy of the compound. The experiment ended on day 25, and mice with a body weight reduction of more than 20% were judged to be dead and euthanized. The experimental procedure is shown in FIG. 2A. The death of mice in each group was recorded, and the survival curve of mice was plotted, as shown in FIG. 2B. The viral load in the plasma of mice in each group is shown in FIGS. 2C and 2D.

Claims

1. A compound represented by the formula (II), a stereoisomer, a tautomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that the fused ring contains one or more heteroatoms; X is H or halogen; R 1 each independently -OC 1-3 alkyl, said -OC 1-3 alkyl optionally substituted with one or more halogen; R 2 H or -OC 1-3 alkyl; R 3 is H or -OC 1-3 alkyl; R 4 is H or -S(=0)2C 1-3 alkyl; n is 0, 1, 2 or 3.

2. The compound, stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to claim 1, wherein, the heteroatom contained in the fused ring is N; and / or the fused ring contains one heteroatom; and / or the ring A is a tricyclic fused aromatic heterocycle, the heteroatom of the tricyclic fused aromatic heterocycle is N; preferably, the number of the heteroatom is 1.

3. The compound, stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to claim 2, wherein, Said ring A comprises the structure of a benzo five-membered nitrogen heterocycle, which is preferably Preferably, ring A is wherein, the ring B is a 4-7 membered heterocycloalkane, a 4-7 membered cycloalkane, a 4-7 membered aromatic ring or a 4-7 membered aromatic heterocycle; More preferably, ring A is Still more preferably 4. The compound, stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to claim 3, wherein, For Preferably 5. The compound, stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to claim 3 or 4, wherein the ring B is a 5-6 membered heterocycloalkane, a 5-6 membered cycloalkane, a 5-6 membered aromatic ring or a 5-6 membered aromatic heterocycle.

6. The compound, stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to claim 1, wherein has any one of the structures shown below: wherein n number of each independently R 1 may optionally substitute the substitutable position of the tricyclic fused ring; Preferably, the compound, stereoisomer, tautomer thereof or pharmaceutically acceptable salt thereof has a structure shown in formula (II-4).

7. The compound, stereoisomer, tautomer or pharmaceutically acceptable salt thereof according to any one of claims 1-6, wherein, R 1 is -OC 1-3 alkyl, said -OC 1-3 alkyl is optionally substituted with one or more F; preferably R 1 is -OCH3, -OCHF2, -OCH2F or -OCF3, more preferably -OCF3; and / or n is 0, 1 or 2, preferably 0 or 1; and / or X is halogen, preferably -Cl; and / or R 2 is -OC 1-3 alkyl, preferably -OCH3; and / or R 3 is -OC 1-3 alkyl, preferably -OCH3; and / or R 4 -S(=O)2C 1-3 alkyl, preferably -S(=O)2CH3.

8. The compound of claim 1, stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein, as represented by any of the following structural formulas: and / or The compound represented by formula (II), a stereoisomer, a tautomer thereof, or a pharmaceutically acceptable salt thereof is represented by any one of the following structural formulas:

9. A pharmaceutical composition comprising at least one compound, stereoisomer, tautomer thereof or pharmaceutically acceptable salt thereof according to any one of claims 1-8, and optionally one or more pharmaceutically acceptable carriers and / or additives.

10. Use of a compound, stereoisomer, tautomer thereof or pharmaceutically acceptable salt thereof according to any one of claims 1-8 or a pharmaceutical composition according to claim 9 in the preparation of a medicament for the prevention or treatment of a flavivirus infection; preferably, the flavivirus infection is a dengue virus or yellow fever virus infection.

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