PRMT5 inhibitor and its use

Abstract

The present invention provides a compound having methyltransferase inhibitory activity. Specifically, the present invention provides a compound having PRMT5 inhibitory activity. The compound can be used for the preparation of a pharmaceutical composition for treating PRMT5 activity-related diseases. Formula I, Formula II.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical compounds. Specifically, the present invention provides compounds that inhibit PRMT5 and their use for pharmaceutical compositions.

Background Art

[0002] Epigenetic control of gene expression is an important biological determinant of protein production and cell differentiation and plays an important pathogenic role in many human diseases. Epigenetic control involves genetically modifying genetic material without altering the nucleotide sequence. Typically, epigenetic control is mediated by selective and reversible modifications (e.g., methylation) of DNA and proteins (e.g., histones), and these modifications control the structural transition between the transcriptionally active and inactive states of chromatin. These covalent modifications are controlled by enzymes such as methyltransferases (e.g., PRMT5), where many are associated with specific gene mutations that can cause human diseases. PRMT5 plays a role in diseases such as proliferative diseases, metabolic diseases, and blood diseases.

[0003] PRMT5 is a known essential cell gene, and conditional PRMT5 knockout and siRNA knockout studies have shown that PRMT5 inhibition in normal tissues is associated with a series of diseases (e.g., pancytopenia, infertility, sarcopenia, cardiac hypertrophy). Therefore, there is a need for new strategies to exploit such metabolic vulnerabilities and preferentially target PRMT5 in MTAP-null tumors (MTAPWT) while sparing PRMT5 in normal tissues. Targeting PRMT5 with MTA-competitive small molecule inhibitors preferentially targets the MTA-binding state of PRMT5, provides a better therapeutic index than normal cells rich in MTAP-null tumor cells and with low MTA levels without compromising MTAP.

[0004] Therefore, in the art, there is a need to provide a new small molecule compound targeting PRMT5 in MTAP-null tumors.

SUMMARY OF THE INVENTION

PROBLEMS TO BE SOLVED BY THE INVENTION

[0005] An object of the present invention is to provide a new small molecule compound targeting PRMT5 in MTAP-null tumors.

MEANS FOR SOLVING THE PROBLEM

[0006] A first aspect of the present invention provides a compound as shown in the following formula I, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof,

CHEMICAL FORMULA

CHEMICAL FORMULA

CHEMICAL FORMULA

Chemical formula

Chemical formula

[0007] In another preferred example, the Ra is selected from the group consisting of the following:

Chemical formula

[0008] In another preferred example, the L1 is -CHR- or -C(R)R-, the A ring is selected from the group consisting of a substituted or unsubstituted 8- to 12-membered fused bicyclic heterocyclic group and a substituted or unsubstituted 7- to 10-membered fused bicyclic heteroaryl group, the R8 is selected from the group consisting of H, halogen, cyano group, amino group, alkynyl group, SF5, hydroxy group, thiol group, aldehyde group, carboxy group, unsubstituted or halogenated C1-C6 alkyl group,

Chemical formula

[0009] In another preferred example, said R2 is selected from the group consisting of R7 and -L2R7, where said L2 is selected from the group consisting of -O-, -CHR-, a carbonyl group, S, and -NH-, and where R7 is a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C 6-10 is selected from the group consisting of an aromatic ring, a substituted or unsubstituted 5- to 12-membered heteroaromatic ring.

[0010] In another preferred example, said R2 is an ortho-substituted 5- or 6-membered heteroaromatic ring as follows: [Chemical formula] The ortho substituent R 10 is selected from the group consisting of hydrogen, deuterium, a halogen, a halogenated or non-halogenated C1-C3 alkyl group, and a halogenated or non-halogenated C1-C3 alkoxy group. The D ring is selected from the group consisting of a substituted or unsubstituted benzene ring and a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, and preferably, the D ring is selected from the following group. [Chemical formula]

[0011] In another preferred example, said L1 is -CH2- or -CH(CH3)-, and the A ring is selected from the following group: [Chemical formula] Here, the C ring is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring (including the case of being saturated or partially unsaturated), and a substituted or unsubstituted 3- to 6-membered heterocyclic ring (including the case of being saturated or partially unsaturated). Or the A ring is selected from the following group: [Chemical formula] R8 is [Chemical formula] and the B ring is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, a substituted or unsubstituted C3-C6 carbocyclic ring, and a substituted or unsubstituted 3- to 6-membered heterocyclic ring, and L3 is selected from the group consisting of a chemical bond, -O-, -CHR-, a carbonyl group, S, or -NH-.

[0012] In another preferred example, the L3 is a chemical bond.

[0013] In another preferred example, the B ring is a substituted or unsubstituted benzene ring or a substituted or unsubstituted 5- to 6-membered heteroaromatic ring.

[0014] In another preferred example, R2 is selected from the group consisting of R7 and -(CHR)R7, where R7 is hydrogen or absent, a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted 6-10 aromatic ring, a substituted or unsubstituted 5- to 12-membered heteroaromatic ring, a substituted or unsubstituted C3-C8 carbocyclic ring (including saturated or partially unsaturated cases, including monocyclic, fused-ring, spiro-ring or bridged-ring), and a substituted or unsubstituted 3- to 8-membered heterocyclic ring (including saturated or partially unsaturated cases, including monocyclic, fused-ring, spiro-ring or bridged-ring), which is selected from the group consisting of.

[0015] In another preferred example, the compound has a structure as shown in the following formula:

Chemical formula

Chemical formula

[0016] In another preferred example, said R3 is selected from the group consisting of H, deuterium, halogen, cyano group, substituted or unsubstituted C1-C6 alkyl group.

[0017] In another preferred example, said R2 is a substituted or unsubstituted 5- to 7-membered heteroaromatic ring, and said A ring is selected from the group consisting of a substituted or unsubstituted 5- to 6-membered aromatic ring or heteroaromatic ring, a substituted or unsubstituted 7- to 10-membered fused bicyclic heteroaryl group, and said R8 is CF3.

[0018] The second aspect of the present invention provides a pharmaceutical composition, said pharmaceutical composition comprising a therapeutically effective amount of one or more compounds according to any of the foregoing aspects, its pharmaceutically acceptable salts, racemic compounds, optical isomers, stereoisomers or tautomers, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary substances and / or diluents.

[0019] The third aspect of the present invention provides the use of a compound according to any of the foregoing aspects, its racemic compound, optical isomer or pharmaceutically acceptable salt in the preparation of a drug for treating or preventing a disease associated with abnormal gene level or abnormal expression of PRMT5 (for example, corresponding nucleic acid mutation, deletion, or generation of methyltransferase ectopia or fusion or overexpression).

[0020] In another preferred example, the disease is selected from the group consisting of the disease or symptomatic ovarian cancer, lung cancer, lymphoma, glioblastoma, colorectal cancer, melanoma, gastric cancer, pancreatic cancer or bladder cancer.

Advantages of the Invention

[0021] It should be understood that within the scope of the present invention, new or preferred technical solutions can be formed by combining each of the above technical features of the present invention with the technical features specifically described below (for example, in the examples) with each other. Due to space limitations, it will not be repeated here.

Modes for Carrying Out the Invention

[0022] Through extensive and meticulous research, the inventors of the present invention unexpectedly discovered for the first time a compound having a PRMT5 regulatory effect. Based on this, the present invention was completed.

[0023] Term

[0024] In the present invention, the halogen is F, Cl, Br or I.

[0025] In the present invention, unless otherwise specified, the terms used have the ordinary meanings known to those skilled in the art. In the present invention, unless otherwise specified, all chemical formulas include all possible optical isomers or geometric isomers (for example, R-type, S-type or racemic, or cis-trans isomers of alkenes, etc.).

[0026] In the present invention, the term "C1-C6 alkyl group" refers to a straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms, and non-limitingly includes methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, s-butyl group, t-butyl group, pentyl group and hexyl group, etc., preferably including ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, s-butyl group and t-butyl group.

[0027] In the present invention, the term "C1-C6 alkoxy group" refers to a linear or branched alkoxy group having 1 to 6 carbon atoms, and includes, without limitation, a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, and the like.

[0028] In the present invention, the term "C2-C6 alkenyl group" refers to a linear or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, and includes, without limitation, a vinyl group, a propenyl group, a butenyl group, an isobutenyl group, a pentenyl group, a hexenyl group, and the like.

[0029] In the present invention, the term "C2-C6 alkynyl group" refers to a linear or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, and includes, without limitation, an ethynyl group, a propynyl group, a butynyl group, an isobutynyl group, a pentynyl group, a hexynyl group, and the like.

[0030] In the present invention, "C3-C 10 cycloalkyl group" refers to a cyclic alkyl group having 3 to 10 carbon atoms in the ring, and includes, without limitation, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, a cyclodecyl group, and the like. The terms "C3-C8 cycloalkyl group", "C3-C7 cycloalkyl group", and "C3-C6 cycloalkyl group" have similar meanings.

[0031] In the present invention, "C3-C 10 cycloalkenyl group" refers to a cyclic alkenyl group having 3 to 10 carbon atoms in the ring, and includes, without limitation, a cyclopropenyl group, a cyclobutenyl group, a cyclopentenyl group, a cyclohexenyl group, a cycloheptenyl group, a cyclooctenyl group, a cyclodecenyl group, and the like. The term "C3-C7 cycloalkenyl group" has a similar meaning.

[0032] In the present invention, "C1-C 12The term "alkoxycarbonyl group" refers to an alkoxycarbonyl group having 1 to 12 carbon atoms on the alkyl chain, and includes, without limitation, methoxycarbonyl group, ethoxycarbonyl group, propoxycarbonyl group, isopropoxycarbonyl group, t-butoxycarbonyl group, benzyloxycarbonyl group, etc.

[0033] In the present invention, "C1-C" 12 The term "alkylaminocarbonyl group" refers to an alkylaminocarbonyl group having 1 to 12 carbon atoms on the alkyl chain, and includes, without limitation, methylaminocarbonyl group, ethylaminocarbonyl group, propylaminocarbonyl group, isopropylaminocarbonyl group, t-butylaminocarbonyl group, benzylaminocarbonyl group, dimethylaminocarbonyl group, etc.

[0034] In the present invention, the terms "aromatic ring" or "aryl group" have the same meaning. Preferably, the "aryl group" is a "C6-C" 12 "aryl group" or a "C6-C" 10 "aryl group". "C6-C" 12 The term "aryl group" refers to an aromatic ring group having 6 to 12 carbon atoms that does not contain a heteroatom in the ring such as a phenyl group or a naphthyl group. "C6-C" 10 The term "aryl group" has the same meaning.

[0035] In the present invention, the terms "aromatic heterocycle" or "heteroaryl group" have the same meaning and refer to a heteroaromatic group containing one or more heteroatoms. The heteroatoms referred to in this specification include oxygen, sulfur and nitrogen. For example, furyl group, thienyl group, pyridyl group, pyrazolyl group, pyrrolyl group, N-alkylpyrrolyl group, pyrimidinyl group, pyrazinyl group, imidazolyl group, tetrazolyl group, etc. The heteroaryl group ring can be condensed with an aryl group, a heterocyclic group or a cycloalkyl group ring, where the ring bonded to the parent structure is a heteroaryl group ring. The heteroaryl group may be optionally substituted or unsubstituted.

[0036] In the present invention, the term "3- to 12-membered heterocyclic group" refers to a saturated or unsaturated 3- to 12-membered ring group containing 1 to 3 heteroatoms selected from oxygen, sulfur, and nitrogen in the ring, such as a dioxolanyl group. The term "3- to 7-membered heterocyclic group" has a similar meaning.

[0037] In the present invention, the term "substituted" refers to the substitution of one or more hydrogen atoms of a specific group by a specific substituent. The specific substituent is the aforementioned corresponding substituent or the substituent appearing in each example. Unless otherwise specified, a certain substituent can have a substituent selected from a specific group at any substitutable position of the group, and the substituents may be the same or different at each position. A cyclic substituent such as a heterocycloalkyl group can be bonded to another ring such as a cycloalkyl group to form a spiro ring system, for example, two rings having a common carbon atom. Those skilled in the art will understand that the combinations of substituents contemplated by the present invention are stable or chemically feasible. The substituents are, for example, C 1-8 alkyl group, C 2-8 alkenyl group, C 2-8 alkynyl group, C 3-8 cycloalkyl group, 3- to 12-membered heterocyclic group, aryl group, heteroaryl group, halogen, hydroxyl group, carboxyl group (-COOH), C 1-8 aldehyde group, C 2-10 acyl group, C 2-10 ester group, C1-C 12 alkoxycarbonyl group, amino group, alkoxy group, C 1-10 sulfonyl group, etc. (not limited thereto).

[0038] For example, when the expression "C 1-8 " or a similar expression is used, it means that the group has 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms.

[0039] For example, when an expression such as "3 to 12 members" or a similar expression is used, it means that the group can have 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms or heteroatoms as ring atoms.

[0040] PRMT5 regulator compound

[0041] The present invention provides a compound having PRMT5 regulatory activity,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0042] Pharmaceutical composition and administration method

[0043] Since the compound of the present invention has excellent methyltransferase inhibitory activity, the compound of the present invention and its various crystalline forms, pharmaceutically acceptable inorganic salts or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compound of the present invention as a main active ingredient can be used for treating, preventing and alleviating related diseases caused by abnormal activity or expression level of methyltransferase (for example, PRMT5).

[0044] The pharmaceutical composition of the present invention contains a compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe and effective amount range. Here, "safe and effective amount" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1 to 2000 mg of the compound of the present invention / dosage form, more preferably 1 to 200 mg of the compound of the present invention / dosage form. Preferably, the said "1 dosage form" is one capsule or tablet.

[0045] "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for use in humans and must have sufficient purity and sufficiently low toxicity. "Compatible" means that each component of the composition can be blended with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween (registered trademark)), wetting agents (e.g., sodium dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0046] The administration route of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and typical administration routes include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0047] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with components such as (a) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (c) humectants, such as glycerin, (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, (e) retardants, such as paraffin, (f) absorption promoters, such as quaternary amine compounds, (g) wetting agents, such as cetyl alcohol and glycerol monostearate, (h) adsorbents, such as kaolin, and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form can also include buffering agents.

[0048] Solid dosage forms, such as tablets, sugar pills, capsules, pills, and granules, can be prepared using coating and shell materials, such as enteric coatings and other materials known in the art. They can include opacifying agents, and the release of the active compound or compounds of such compositions can be delayed in a specific part of the digestive tract. Examples of embeddable components that can be used are polymeric substances and waxes. Optionally, the active compound can form microcapsules with one or more of the above excipients.

[0049] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage forms may include inert diluents conventionally used in the art such as water or other solvents, and solubilizing and emulsifying agents such as, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 - butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0050] In addition to these inert diluents, the compositions may also include adjuvants such as, for example, wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes. In addition to the active compound, the suspension may include suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these substances.

[0051] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or non - aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non - aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0052] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required as necessary.

[0053] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds. In some preferred embodiments, the compounds of the present invention can also form PROTACs together with other small molecule compounds or form ADCs for administration together with other macromolecular compounds such as monoclonal antibodies.

[0054] When a pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is applied to a mammal (e.g., human) in need of treatment, where the dosage at the time of administration is the effective dosage to be considered. For a person weighing 60 kg, the daily dose is usually 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dosage needs to take into account factors such as the administration route and the health status of the patient, and all of these are within the scope of the skills of a skilled physician.

[0055] Hereinafter, the present invention will be further described in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and do not limit the scope of the present invention. In the following examples, experimental methods without indicating specific conditions usually follow conventional conditions or conditions proposed by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0056] The definitions of each abbreviation are as follows.

Table 1

[0057] Purification of the intermediates and compounds is carried out by conventional chemical experimental operations such as normal-phase or reverse-phase chromatography or recrystallization. Normal-phase chromatography is preparative silica gel chromatography column or preparative thin-layer chromatography. The silica gel chromatography column is mainly a glass column or high-speed preparative chromatography. The mobile phase of normal-phase chromatography is selected from petroleum ether / ethyl acetate, dichloromethane / methanol or other appropriate solvents and eluted in ratios. Reverse-phase preparative liquid chromatography employs a C18 column, uses preparative liquid chromatography or high-speed preparative chromatography, detects with 214 nM and 254 nM or preparative liquid chromatography-mass spectrometer, and gradient elutes using water / acetonitrile containing 0.1% hydrochloric acid, water / acetonitrile, water / acetonitrile containing 0.1% ammonium bicarbonate, water / acetonitrile containing 0.1% formic acid, water / acetonitrile containing 0.1% aqueous ammonia, water / acetonitrile containing 0.1% trifluoroacetic acid or other appropriate solvent systems as the mobile phase.

[0058] Characterization of the structures of the intermediates and compounds uses the methods of nuclear magnetic resonance (NMR) and mass spectrometry (LCMS). The NMR spectrometers used in nuclear magnetic resonance are Bruker Ascend 400 or Varian 400 or ZKNJ BIXI-1 300 MHz or Bruker Avance III 400 MHz or Bruker AVANCE Neo 400 MHz. The solvents used are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol or other marked deuterated solvents. The spectral data are reported in the mode of chemical shift δ (number of peak splits, coupling constant J (Hz), number of hydrogens). Tetramethylsilane is used as an internal standard substance for chemical shift, and its chemical shift is set to zero (δ, 0 ppm). The meanings of some abbreviations are s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad peak).

[0059] Typical methods of liquid chromatography-mass spectrometry (LCMS) in characterizing the structures of intermediates and compounds are as follows.

[0060] Method 1: It is carried out on an Agilent LC1260 system coupled to a 6120 single quadrupole mass spectrometer. Column: Waters CORTECS C-18, 2.7 μm, 4.6×30 mm. Solvent A: 0.05% aqueous formic acid solution, Solvent B: acetonitrile solution of 0.05% formic acid, from 5% acetonitrile to 95% acetonitrile in 1 minute, maintained for 1 minute, total 2.5 minutes, flow rate: 1.8 mL / min, column temperature 40 °C. Column: XSelect CSH C18, 3.5 μm, 4.6×50 mm. Solvent A: 0.05% aqueous ammonia solution, Solvent B: acetonitrile solution of 0.05% ammonia, from 5% acetonitrile to 95% acetonitrile within 1 minute, maintained for 1 minute, total 2.5 minutes, flow rate: 1.8 mL / min, column temperature 40 °C.

[0061] Method 2: It is carried out on an Agilent LC / MSD 1200 system coupled to a quadrupole mass spectrometer. Column: ODS 2000 (50×4.6 mm, 5 μm) (ES(+) or (-) ionization mode), temperature 30 °C, flow rate 1.5 mL / min.

[0062] General method for the synthesis of examples:

Chemical formula

[0063] General method: Synthesis of intermediate A2 Synthesis route:

Chemical formula

[0064] Step 1: Methyl 2,5-difluoro-4-nitrobenzoate (2) 2,5-Difluoro-4-nitrobenzoic acid (1) (50 g, 246.18 mmol, 1 equiv) is dissolved in methanol (500 mL), and thionyl chloride (43.93 g, 369.28 mmol, 26.79 mL, 1.5 equiv) is added at 0 °C. The reaction mixture is reacted at 40 °C for 16 h. Completion of the reaction is indicated by LCMS. The reaction mixture is concentrated to dryness under reduced pressure. After dilution by adding 300 mL of water, it is extracted three times with 1 L of ethyl acetate. The organic phase is washed with saturated brine (400 mL), dried over anhydrous magnesium sulfate, the filtrate is filtered, and concentrated to dryness under reduced pressure. The crude product is slurried with petroleum ether at 25 °C for 60 min to obtain methyl 2,5-difluoro-4-nitrobenzoate (2) as a white solid (103 g, 474.38 mmol, 96.35% yield). HNMR: ES23714-64-P1A, 1 H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.89 (td, J = 9.51, 5.63 Hz, 2H) 4.00 (s, 3H). 19FnmR (376 MHz, CHLOROFORM-d) δ ppm -110.27 (s, 1F) -121.56 (m, 1F).

[0065] Step 2: Methyl 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3) Methyl 2,5-difluoro-4-nitrobenzoate (2) (80 g, 368.45 mmol, 1 equiv), 2-methyl-1H-imidazole (36.30 g, 442.14 mmol, 1.2 equiv) are dissolved in dimethyl sulfoxide (1.2 L). The reaction solution is reacted at 50 °C for 16 h. Completion of the reaction is indicated by LCMS. After adding 4 L of water to the reaction solution for dilution, extraction is carried out with 4.5 L of ethyl acetate. The organic phase is washed with saturated brine (3 L), dried over anhydrous magnesium sulfate, the filtrate is filtered, and concentrated to dryness under reduced pressure. The crude product is slurried with methyl t-butyl ether at 25 °C for 60 min (25.5 g). (2) The mother liquor is purified by column chromatography (silica, 50% tetrahydrofuran in petroleum ether) to obtain a yellow liquid, which is slurried with methyl t-butyl ether at 25 °C for 60 min to obtain a yellow solid (8.23 g). Yellow solid methyl 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3) (25.5 g, 91.32 mmol, 24.79% yield), methyl 2,5-difluoro-4-nitrobenzoate (2) (recovery of raw material) (20.34 g, 93.68 mmol, 25.43% yield). Yellow solid methyl 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3) (8.2 g, 29.07 mmol, 7.89% yield, 99% purity). HNMR:ES23714-67-P1A1, 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.39 (d, J = 9.90 Hz, 1H) 8.08 - 8.20 (m, 1H) 7.23 (d, J = 1.32 Hz, 1H) 6.92 (s, 1H) 3.91 (s, 3H) 2.12 (s, 3H). 19FnmR (376 MHz, DMSO-d6) δ ppm -105.45 (br s, 1F)

[0066] Step 3: Methyl 4-amino-2-fluoro-5-(2-methyl-1H-imidazol-1-yl)benzoate (4) Methyl 2-fluoro-5-(2-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3) (13.9 g, 49.78 mmol, 1 eq) was dissolved in tetrahydrofuran (300 mL), and palladium on carbon hydroxide (2.8 g, 49.78 mmol, 20% purity, 1 eq) was added under a hydrogen gas atmosphere. The reaction system was replaced with hydrogen gas three times. The reaction solution was heated to 50 °C under a hydrogen gas (1 eq) (50 psi) atmosphere and reacted for 32 hours. Completion of the reaction was indicated by LCMS. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed four times with 300 mL of ethyl acetate. The filtrate was concentrated to dryness under reduced pressure to obtain a gray solid. The crude product was used directly in the next step. Methyl 4-amino-2-fluoro-5-(2-methyl-1H-imidazol-1-yl)benzoate (4) (12 g, 48.15 mmol, 96.72% yield). 1 H NMR (400 MHz, DMSO-d6) Shift 7.46 (d, J = 7.63 Hz, 1H), 7.07 (d, J = 1.38 Hz, 1H), 6.93 (d, J = 1.25 Hz, 1H), 6.59 (d, J = 13.51 Hz, 1H), 6.15 (br s, 2H), 3.68 - 3.77 (m, 3H), 2.01 - 2.12 (m, 3H). 19F nmR (376 MHz, DMSO-d6) Shift -109.31--108.47 (m, 1F).

[0067] Step 4: Methyl 7-fluoro-1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) At 25 °C, methyl 4-amino-2-fluoro-5-(2-methyl-1H-imidazol-1-yl)benzoate (4) (12 g, 48.15 mmol, 1 equivalent) is added to 1-methyl-2-pyrrolidone, and 1,1-carbonyldiimidazole (19.52 g, 120.37 mmol, 2.5 equivalents) is added. The reaction mixture is heated to 115 °C and reacted for 16 hours. Completion of the reaction is indicated by LCMS. The reaction mixtures of two batches are combined and processed. 600 mL of ethyl acetate and 600 mL of water are added to the reaction mixture, and it is slurried at 25 °C for 16 hours. The slurry is filtered under reduced pressure, and the filter cake is washed with 100 mL of ethyl acetate. The solid is concentrated under reduced pressure to obtain gray solid methyl 7-fluoro-1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (24.1 g, 87.56 mmol, 86.60% yield). The crude product is used directly in the next-step reaction. 1 H NMR (400 MHz, DMSO-d6) Shift 11.71 (br s, 1H), 8.40 (d, J = 6.38 Hz, 1H), 7.76 (s, 1H), 7.08 (d, J = 11.38 Hz, 1H), 3.88 (s, 3H), 2.89 (s, 3H). 19FnmR (376 MHz, DMSO-d6) Shift -111.43--110.58 (m, 1F)

[0068] Step 5: Methyl 4-((2,4-dimethoxybenzyl)amino)-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) Methyl 7-fluoro-1-methyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (12.0 g, 43.60 mmol, 1.0 equiv), 2,4-dimethoxybenzylamine (10.9 g, 65.19 mmol, 9.82 mL, 1.50 equiv), and 1,8-diazabicyclo[5.4.0]undec-7-ene (19.92 g, 130.80 mmol, 19.72 mL, 3.0 equiv) are added to acetonitrile (240 mL). Benzotriazol-1-oxo-tris(dimethylaminophosphine) hexafluorophosphate (25.07 g, 56.68 mmol, 1.3 equiv) is added in one batch at 15 - 20 °C. The reaction mixture exotherms slightly, the reaction mixture becomes homogeneous, and a solid precipitates. The reaction mixture is reacted at 15 - 20 °C for 16 h under nitrogen gas protection. LCMS indicates complete consumption of the starting material and detection of the target compound. The reaction suspension is filtered under reduced pressure, and the filter cake is washed with 100 mL of acetonitrile. The solid is collected and dried by suction under reduced pressure to obtain the off-white solid methyl 4-((2,4-dimethoxybenzyl)amino)-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (15.3 g, 36.05 mmol, 82.68% yield). LCMSES15882-1146-P1A: (ESI) m / z = 425.3 [M+1] + , RT = 1.721 min. 1 H NMR (400 MHz, DMSO-d6) Shift 8.49 (d, J = 7.00 Hz, 1H), 8.45 (t, J = 5.57 Hz, 1H), 7.95 (s, 1H), 7.23 (d, J = 12.51 Hz, 1H), 7.18 (d, J = 8.38 Hz, 1H), 6.58 (d, J = 2.38 Hz, 1H), 6.47 (dd, J = 2.38, 8.38 Hz, 1H), 4.66 (d, J = 5.25 Hz, 2H), 3.88 (s, 3H), 3.82 (s, 3H), 3.73 (s, 3H), 2.93 (s, 3H). 19FnmR (376.5 MHz, DMSO-d6) Shift -113.02.

[0069] Step 6: Methyl 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (7) Methyl 4-((2,4-dimethoxybenzyl)amino)-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (16.3 g, 38.40 mmol, 1.0 eq) is added to dichloromethane (50 mL), and trifluoroacetic acid (250 mL) is added. The reaction mixture is heated to 50 °C and reacted for 16 hours. LCMS shows complete consumption of the starting material and detection of the target compound. The reaction mixture is concentrated to dryness under reduced pressure to obtain a purple solid, methyl 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (7) (28.3 g, crude product), and the crude product is used directly in the next step reaction. LCMSES15882-1150-P1A: (ESI) m / z = 275.3 [M+1] + , RT = 0.607 min

[0070] Step 7: 4-Amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A2) Methyl 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (7) (crude product of the previous step) (28.3 g, 38.70 mmol, 1 equivalent) is added to tetrahydrofuran (80 mL) and methanol (80 mL). Sodium hydroxide (7.74 g, 193.48 mmol, 5 equivalents) is dissolved in water (80 mL) and then added to the reaction solution. The reaction solution is heated to 50 °C and reacted for 4 hours. The required compound is detected by LCMS. After the reaction solution is cooled to 20 °C, it is concentrated under reduced pressure to remove the organic solvent, and the residue is diluted with 10-to-1 water-to-methanol (300 mL), then filtered through diatomaceous earth, and the filter cake is washed repeatedly 3 times with 10-to-1 water-to-methanol (300 mL). All the filtrates are combined and concentrated under reduced pressure to remove methanol. The pH of the residue is adjusted to 5 - 6 with acetic acid. The resulting slurry is stirred at 15 - 20 °C for 12 hours and filtered under reduced pressure, and the obtained solid is washed with water. The solid is collected and freeze-dried to obtain a white solid, 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A2) (9.9 g, 37.55 mmol, 97.05% yield, 98.71% purity). LCMSES15882-1154-P1C: (ESI) m / z = 261.1 [M+1] + , RT = 0.422 min. 1 H NMR (400 MHz, DMSO-d6) Shift 13.15 (br s, 1H), 8.53 (d, J = 7.04 Hz, 1H), 7.85 (s, 1H), 7.68 (s, 2H), 7.16 (d, J = 12.10 Hz, 1H), 2.94 (s, 3H)

[0071] General method: Synthesis of Intermediate A3 Synthetic route:

Chemical formula

[0072] Step 1: 1-(5-Bromo-4-chloro-2-nitro-phenyl)-2-methyl-imidazole (2) 1-Bromo-2-chloro-5-fluoro-4-nitrobenzene (1) (3 g, 11 mmol, 1 equiv) was added to a solution of 2-methyl-1H-imidazole (1.2 g, 14 mmol, 1.2 equiv) in acetonitrile (50 mL), and then potassium carbonate (4 g, 29 mmol, 2.5 equiv) was added to the reaction mixture. The temperature was raised to 80 °C and the mixture was stirred for 16 h. The completion of the reaction of the starting materials was detected and the target product was formed. The mixture was concentrated under reduced pressure to remove acetonitrile, water (40 mL) was added, and the mixture was extracted with ethyl acetate (3 × 50 mL). The extract was dried over anhydrous magnesium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography (silica, 35% tetrahydrofuran in petroleum ether) to obtain 1-(5-bromo-4-chloro-2-nitro-phenyl)-2-methyl-imidazole (2) (4 g) as a white solid. HnmR: 1 H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.32 (s, 1H), 7.22 (d, J = 1.32 Hz, 1H), 6.91 (d, J = 1.32 Hz, 1H), 2.08 - 2.23 (m, 3H).

[0073] Step 2: 4-Bromo-5-chloro-2-(2-methylimidazol-1-yl)aniline (3) 1-(5-Bromo-4-chloro-2-nitro-phenyl)-2-methyl-imidazole (2) (3.5 g, 11 mmol, 1 equiv) was dissolved in a mixed solution of (8 mL), ethanol (16 mL) and tetrahydrofuran (16 mL), ammonium chloride (8.9 g, 166 mmol, 15 equiv) was added to the reaction mixture, the temperature was raised to 70 °C, and iron powder (2.5 g, 44 mmol, 4 equiv) was added to the reaction mixture. The reaction mixture was stirred at 90 °C for 2 h. The completion of the reaction of the starting materials was detected by LC-MS and the target product was formed. The reaction mixture was filtered through diatomaceous earth and washed with ethyl acetate (40 mL × 3). The solvent was removed under reduced pressure to obtain 4-bromo-5-chloro-2-(2-methylimidazol-1-yl)aniline (3) (3.1 g, 10.8 mmol, 98% yield) as a black solid. HnmR: 11H NMR (400 MHz, DMSO-d6) δ 7.41 (s, 1H), 7.12 (s, 1H), 7.07 (s, 1H), 6.99 (s, 1H), 5.45 (s, 2H), 2.13 (s, 3H).

[0074] Step 3: 8-Bromo-7-chloro-1-methyl-5H-imidazo[1,5-a]quinoxalin-4-one (4) 4-Bromo-5-chloro-2-(2-methylimidazol-1-yl)aniline (3) (3 g, 10.5 mmol, 1 equiv) and 1,1-carbonyldiimidazole (2.6 g, 15.7 mmol, 1.5 equiv) are successively dissolved in a solution of 1,2-dichlorobenzene (30 mL). The reaction solution is stirred at 130 °C for 16 h. The completion of the reaction of the raw materials is detected by LC-MS, and the target product is formed. The reaction solution is stirred with a solution of ethyl acetate and water (2 / 1 15 mL) for 30 min and then filtered to obtain a filter cake. The filter cake is concentrated under vacuum to obtain a black solid, 8-bromo-7-chloro-1-methyl-5H-imidazo[1,5-a]quinoxalin-4-one (4) (2.2 g, 7 mmol, 67% yield). HnmR: 1 1H NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.33 (d, J = 3.30 Hz, 2H), 2.83 (s, 3H)

[0075] Step 4: Methyl 7-chloro-1-methyl-4-oxo-5H-imidazo[1,5-a]quinoxaline-8-carboxylate (5) 8-Bromo-7-chloro-1-methyl-5H-imidazo[1,5-a]quinoxalin-4-one (4) (500 mg, 1.6 mmol, 1 equiv) was dissolved in an ethanol (5 mL) solution, 1,8-diazabicyclo[5.4.0]undec-7-ene (365 mg, 2.4 mmol, 362 μL, 1.5 equiv) was added, the nitrogen gas was replaced three times, and tributylphosphonium tetrafluoroborate (46 mg, 160 μmol, 0.1 equiv), molybdenum hexacarbonyl (232 mg, 880 μmol, 118 μL, 0.55 equiv) and palladium acetate (36 mg, 160 μmol, 0.1 equiv) were added. The reaction mixture was stirred at 90 °C for 2 h. The completion of the reaction of the raw materials was detected by LC-MS, and the target product was formed. Ethanol was removed under reduced pressure from the reaction solution, water (10 mL) was added, and extraction was performed with 30 mL (10 mL × 3) of ethyl acetate. It was dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by column chromatography (silica, 30% tetrahydrofuran in petroleum ether) gave 7-chloro-1-methyl-4-oxo-5H-imidazo[1,5-a]quinoxaline-8-carboxylic acid methyl ester (5) (500 mg) as a yellow solid.

[0076] Step 5: Methyl 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) Methyl 7-chloro-1-methyl-4-oxo-5H-imidazo[1,5-a]quinoxaline-8-carboxylate (5) (480 mg, 1.6 mmol, 1 equiv) was dissolved in acetonitrile (5 mL), and 2,4-dimethoxybenzylamine (341 mg, 2 mmol, 306 μL, 1.3 equiv), benzotriazol-1-oxo-tris(dimethylaminophosphine) hexafluorophosphate (1 g, 2.4 mmol, 1.5 equiv) and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.2 g, 7.9 mmol, 1.2 mL, 5 equiv) were added. The reaction mixture was stirred at room temperature for 16 h. Completion of the reaction of the starting material was detected by LC-MS, and the desired product was formed. Acetonitrile was removed under reduced pressure from the reaction mixture, water (8 mL) was added, and the mixture was extracted with ethyl acetate (30 mL, 10 mL×3). The extract was dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. Purification by column chromatography (silica, 30% tetrahydrofuran in petroleum ether) gave the yellow solid methyl 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (500 mg, 1.1 mmol, 70% yield).

[0077] Step 6: 7-Chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A3) Methyl 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (500 mg, 1.1 mmol, 1 equiv) is dissolved in water (5 mL) and ethanol (5 mL), and sodium hydroxide (132 mg, 3.3 mmol, 3 equiv) is added. The reaction mixture is stirred at 50 °C for 5 h. Completion of the reaction of the starting material is detected by LCMS, and the desired product is formed. 6 M hydrochloric acid (0.5 mL) is added to the reaction mixture, and the mixture is concentrated to dryness under reduced pressure. The crude product is used directly in the next step to obtain the white solid 7-chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A3) (350 mg, 820 μmol, 74.6% yield).

[0078] General method: Synthesis of Intermediate A4 Synthetic route:

Chemical formula

[0079] Step 1: Methyl 3-(2,4-dimethylimidazol-1-yl)-4-nitrobenzoate Methyl 3-fluoro-4-nitrobenzoate (1) (2.00 g, 10.04 mmol, 1 equiv) is added to acetonitrile (40 equiv), potassium carbonate (4.16 g, 30.13 mmol, 3 equiv) and 2,4-dimethyl-1H-imidazole (2) (965 mg, 10.04 mmol, 1 equiv) are added, and the reaction mixture is reacted at 85 °C for 16 h. Complete consumption of the starting material is indicated by LC-MS, and the desired product is formed. The reaction mixture is concentrated to dryness under reduced pressure, the residue is diluted with dichloromethane (80 equiv), filtered, the filtrate is dried over magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain the yellow solid methyl 3-(2,4-dimethylimidazol-1-yl)-4-nitrobenzoate (3) (2.6 g, 9.45 mmol, 94.05% yield), which is used directly in the next step without further purification as the crude product.

[0080] Step 2: Methyl 4-amino-3-(2,4-dimethylimidazol-1-yl)benzoate Methyl 3-(2,4-dimethylimidazol-1-yl)-4-nitrobenzoate (3) (2.5 g, 9.08 mmol, 1 equiv) is dissolved in ethanol (20 equiv), tetrahydrofuran (20 equiv) and water (10 equiv), and iron powder (5.07 g, 90.82 mmol, 10 equiv) and ammonium chloride (2.43 g, 45.41 mmol, 5 equiv) are added at room temperature. The reaction mixture is reacted at 90 °C for 16 h. Complete consumption of the starting material is indicated by LC-MS, and the target product is formed. The reaction mixture is filtered, concentrated to dryness under reduced pressure, the residue is diluted with water (50 equiv), extracted with ethyl acetate (40 equiv × 3), the organic phases are combined, dried over magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain the yellow solid methyl 4-amino-3-(2,4-dimethylimidazol-1-yl)benzoate (4) (1.9 g, 6.13 mmol, 67.49% yield, 79.13% purity), and the crude product is used directly in the next step without further purification. LCMS: ES19974-375-P1C2, (ESI) m / z = 246.1 [M+1] + , RT = 0.61 min, purity: 79.13%

[0081] Step 3: Methyl 1,3-dimethyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate Methyl 4-amino-3-(2,4-dimethylimidazol-1-yl)benzoate (4) (500 mg, 2.04 mmol, 1 equiv) and 1,1-carbonyldiimidazole (495 mg, 3.06 mmol, 1.5 equiv) were dissolved in 1,2-dichlorobenzene (10 equiv), and the reaction mixture was reacted at 120 °C for 16 h under nitrogen gas protection. Complete consumption of the starting material was indicated by LC-MS, and the target product was formed. The reaction mixture was suction filtered, the filter cake was slurried with water, and dried under reduced pressure to obtain a brown solid, methyl 1,3-dimethyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (460 mg, 1.62 mmol, 79.48% yield, 95.55% purity). Without further purification, the crude product was used directly in the next step. LCMS: ES19974-392-P1B1, (ESI) m / z = 272.0 [M+1] + , RT = 0.61 min, purity: 95.55%

[0082] Step 4: Methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylate Methyl dimethyl-4-oxo-4,5-dihydroimidazo[1,5-a]quinoxaline-8-carboxylate (5) (700 mg, 2.58 mmol, 1 equiv) was dissolved in acetonitrile (25 equiv), and benzotriazol-1-oxo-tris(dimethylaminophosphine) hexafluorophosphate (1.83 g, 4.13 mmol, 1.6 equiv) and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.96 g, 12.90 mmol, 1.94 equiv, 5 equiv) were added. The reaction mixture was reacted at room temperature for 30 minutes, and (2,4-dimethoxyphenyl)methylamine (647.2 mg, 3.87 mmol, 583.1 μL, 1.5 equiv) was added to the reaction mixture. The reaction mixture was reacted at 50 °C for 15.5 hours. Complete consumption of the starting material was indicated by LC-MS, and the target product was formed. The reaction mixture was filtered, and the filter cake was dried under reduced pressure to obtain a brown solid, methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (900 mg, 2.08 mmol, 80.57% yield, 97.13% purity). Without further purification, the crude product was used directly in the next step. LCMS: ES19974-397-P1B1, (ESI) m / z = 421.1 [M+1] + , RT = 0.78 min, purity: 97.13%

[0083] Step 5: 4-((2,4-Dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid Methyl 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylate (6) (850 mg, 2.02 mmol, 1 equiv) was dissolved in methanol (10 equiv), tetrahydrofuran (10 equiv) and water (5 equiv), and lithium hydroxide (424.2 mg, 10.11 mmol, 5 equiv) was added. The reaction mixture was reacted at 50 °C for 16 h. LC-MS indicated complete consumption of the starting material and formation of the desired product. The reaction mixture was concentrated to dryness under reduced pressure, acetic acid was added to the residue to adjust the pH to 6 - 7, filtered, and the filter cake was dried under reduced pressure to obtain 4-((2,4-dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A4) (800 mg, 1.97 mmol, 97.37% yield) as a brown solid. Without further purification, the crude product was used directly in the next step.

[0084] General method: Synthesis of Intermediate A6 Synthetic route:

Chemical formula

[0085] Step 1: N-(4-Bromo-2-fluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (3) A solution of 4-bromo-2-fluoroaniline (5.78 g, 30.4 mmol, 1 eq) and 4-methyl-1H-pyrazole-5-carboxylic acid (4.60 g, 36.5 mmol, 1.2 eq) in Py (120 mL) was added with POCl3 (4.66 g, 30.4 mmol, 2.82 mL, 1 eq), and stirred at 0 °C for 1 h. LC-MS (ET63399-4-R1A1) indicated complete consumption of the starting materials and detected the major peak with the required m / z. The reaction mixture was quenched with ice water (200 mL) and then extracted with ethyl acetate (60 mL×6). The combined organic layers were washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The crude product was triturated with ethyl acetate (50.0 mL) to afford N-(4-bromo-2-fluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (3) as a white solid (7.09 g, 23.8 mmol, 78.2% yield). 1 H NMR (400 MHz, DMSO-d6) δ 13.26 (br s, 1H) 9.52 (s, 1H) 7.92 (t, J = 8.52 Hz, 1H) 7.70 (s, 1H) 7.62 (dd, J = 10.31, 1.85 Hz, 1H) 7.41 (br d, J = 8.70 Hz, 1H) 2.25 (s, 3H). LC-MS; [MH] + 298.0

[0086] Step 2: 8-Bromo-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (4) NaH (1.43 g, 35.7 mmol, 60% purity, 1.5 eq) was added to a solution of N-(4-bromo-2-fluorophenyl)-4-methyl-1H-pyrazole-5-carboxamide (3) (7.09 g, 23.8 mmol, 1 eq) in DMA (70 mL). The mixture was stirred at 120 °C for 16 h. LC-MS indicated complete consumption of the starting materials and the major peak had the required m / z. The reaction mixture was quenched with saturated ammonium chloride (300 mL), the precipitate was collected, washed with water (50 mL), and then concentrated under reduced pressure to give 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (4) as a white solid (6.50 g, crude product).1 1H NMR (400 MHz, DMSO-d6) δ 8.11 (d, J = 2.00 Hz, 1H) 7.91 (s, 1H) 7.52 (dd, J = 8.63, 2.00 Hz, 1H) 7.28 (d, J = 8.63 Hz, 1H) 2.42 (s, 3H). LC-MS; [MH] + 278.0

[0087] Step 3: 8-Bromo-N-(4-methoxybenzyl)-3-methyl-4,5-dihydropyrazolo[1,5-a]quinoxalin-4-amine (5) To a solution of 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4(5H)-one (4) (1.00 g, 3.60 mmol, 1 equiv) in MeCN (10.0 mL) is added PMBNH2 (1.23 g, 8.99 mmol, 1.16 mL, 2.5 equiv), BOP (3.18 g, 7.19 mmol, 2 equiv)) and DBU (2.74 g, 18.0 mmol, 2.71 mL, 5 equiv). The mixture is stirred at 50 °C for 16 h. LC-MS shows complete consumption of the starting material and the required m / z for one major peak. The reaction mixture is diluted with saturated ammonium chloride (10.0 mL) and ethanol (5.00 mL). The precipitate is collected, washed with water (10.0 mL), and then concentrated under reduced pressure to give 8-bromo-N-(4-methoxybenzyl)-3-methyl-4,5-dihydropyrazolo[1,5-a]quinoxalin-4-amine (5) (1.09 g, crude product) as a yellow solid. 1 1H NMR (400 MHz, DMSO-d6) δ 8.20 (d, J = 1.88 Hz, 1H) 7.95 (s, 1H), 7.44 - 7.48 (m, 2H) 7.38 (d, J = 8.63 Hz, 2H) 7.28 (br s, 1H) 6.87 (d, J = 8.75 Hz, 2H) 4.70 (d, J = 5.88 Hz, 2H) 3.71 (s, 3H) 2.53 (s, 3H). LC-MS; [MH] + 399.0

[0088] Step 4: 8-Bromo-3-methylpyrazolo[1,5-a]quinoxalin-4-amine (6) A solution of 8-bromo-N-(4-methoxybenzyl)-3-methyl-4,5-dihydropyrazolo[1,5-a]quinoxalin-4-amine (5) (500 mg, 1.26 mmol, 1 equiv) in TFA (5 mL) is stirred at 60 °C for 12 h. Complete consumption of the starting material is indicated by LC-MS, and the required m / z is present in one major peak. The reaction mixture is concentrated under reduced pressure to afford the compound 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4-amine (6) (400 mg, crude product) as a yellow solid. LC-MS: [MH] + 277.0

[0089] Step 5: Ethyl 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylate (7) To a solution of 8-bromo-3-methylpyrazolo[1,5-a]quinoxalin-4-amine (6) (400 mg, 1.08 mmol, 1 equiv) in EtOH (4 mL) are added Mo(CO)6 (143 mg, 541 μmol, 72.9 μL, 0.5 equiv), DBU (659 mg, 4.33 mmol, 652 μL, 4 eq), (t-Bu)3PBF4 (94.2 mg, 325 μmol, 0.3 eq) and Pd(OAc)2 (36.5 mg, 162 μmol, 0.15 eq). The mixture is stirred at 90 °C for 12 h. Complete consumption of the starting material is indicated by LC-MS (ET63399-28-R1A1), and the required mass is detected. The reaction mixture is diluted with saturated ammonium chloride (10 mL) and ethanol (10 mL). The precipitate is collected and washed with water (10.0 mL) and then concentrated under reduced pressure to afford the yellow solid ethyl 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylate (7) (250 mg, 925 μmol, 85.4% yield). LC-MS; [MH] + 271.1

[0090] Step 6: 4-Amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A6) To a solution of ethyl 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylate (7) (250 mg, 925 μmol, 1 equiv) in EtOH (3 mL) and H2O (1 mL) is added LiOH·H2O (116 mg, 2.77 mmol, 3 equiv). The mixture is stirred at 25 °C for 12 h. LC-MS (ET63399-32-R1A1) indicates complete consumption of the starting material and the required mass is detected. The reaction mixture is concentrated, diluted with water (10 mL), and washed with DCM (10.0 mL × 3) to remove impurities. The aqueous phase is adjusted to pH = 5 with 1M HCl, the precipitate is collected, and purified by preparative HPLC (column: Phenomenex Luna C18 75×30 mm×3 μm, mobile phase: [water (FA)-ACN], B%: 1% - 35%, 8 min, UV220 nm and 254 nm) to afford 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A6) as a yellow solid (60.0 mg, 248 μmol, 26.8% yield). 1 1H NMR (400 MHz, DMSO-d6) δ 12.94 (br s, 1H) 8.67 (d, J = 1.96 Hz, 1H) 7.96 (s, 1H) 7.89 (dd, J = 8.44, 1.96 Hz, 1H) 7.52 (d, J = 8.44 Hz, 1H) 7.19 (br s, 2H) 2.49 (br s, 3H). LC-MS, [M+H] + 243.1

[0091] General method: Synthesis of Intermediate A7 Synthetic route:

Chemical Structure

[0092] Step 1: Methyl 2-fluoro-5-(4-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (3) A mixture of methyl 2,5-difluoro-4-nitrobenzoate (20.0 g, 92.1 mmol, 1 equiv) and 5-methyl-1H-imidazole (7.56 g, 92.1 mmol, 1 equiv) in DMSO (200 mL) is stirred at 50 °C for 12 h. Complete consumption of the starting material is indicated by LCMS and the required mass is detected. The residue is diluted with H2O (400 mL) and extracted with EtOAc (200 mL × 3). The combined organic layers are washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue is purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 0 / 1) to give methyl 2-fluoro-5-(4-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (9.30 g, 33.3 mmol, 34.2% yield) as a yellow solid. LC-MS (ESI) m / z = 280.1 [M+H] +

[0093] Step 2: Methyl 4-amino-2-fluoro-5-(4-methyl-1H-imidazol-1-yl)benzoate (4) Iron powder (7.44 g, 133 mmol, 4 equiv) is added to a solution of methyl 2-fluoro-5-(4-methyl-1H-imidazol-1-yl)-4-nitrobenzoate (9.30 g, 33.3 mmol, 1 equiv) and NH4Cl (26.7 g, 499 mmol, 15 equiv) in EtOH (90.0 mL) / THF (90.0 mL) / H2O (45.0 mL). The mixture is stirred at 80 °C for 2 h. Complete consumption of the starting material is indicated by LCMS and the required mass is detected. The reaction mixture is filtered, the filtrate is diluted with water (300 mL), and extracted with EtOAc (200 mL × 3). The combined organic layers are washed with brine (100 mL), dried over Na2SO4, and concentrated under reduced pressure to give methyl 4-amino-2-fluoro-5-(4-methyl-1H-imidazol-1-yl)benzoate (8.00 g, 32.1 mmol, 95.2% yield) as a yellow solid. LC-MS (ESI) m / z = 250.0 [M+H] +

[0094] Step 3: Methyl 7-fluoro-4-hydroxy-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate (5) Methyl 4-amino-2-fluoro-5-(4-methyl-1H-imidazol-1-yl)benzoate (500 mg, 2.01 mmol, 1 eq) and CDI (487 mg, 2.41 mmol, 1.2 eq) are placed in 1,2-dichlorobenzene (20.0 mL) in a microwave tube. The sealed tube is heated at 150 °C for 3 h under microwave. 11 parallel reactions are carried out. Complete consumption of the starting material is indicated by LCMS and the required mass is detected. The reaction mixture is filtered and the filter cake is dried under reduced pressure to give a residue. The crude product and MTBE (20.0 mL) are triturated at 25 °C for 30 min to give methyl 7-fluoro-4-hydroxy-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate as a yellow solid (6.00 g, 21.80 mmol, 85.0% yield). LC-MS (ESI) m / z = 276.0 [M+H] +

[0095] Step 4: Methyl 7-fluoro-4-((4-methoxybenzyl)amino)-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) A solution of methyl 7-fluoro-4-hydroxy-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate 5 (6.00 g, 21.8 mmol, 1 equiv) in MeCN (120 mL) is added to BOP (19.2 g, 43.6 mmol, 2 equiv), DBU (16.5 g, 109 mmol, 16.4 mL, 5 equiv)) and PMBNH2 (7.48 g, 54.5 mmol, 7.05 mL, 2.5 equiv). The mixture is stirred at 70 °C for 16 h. Complete consumption of the starting material is indicated by LCMS and the required mass is detected. The reaction mixture is diluted with saturated ammonium chloride (20.0 mL). The filter cake is washed with water (10.0 mL) and concentrated under reduced pressure to afford methyl 7-fluoro-4-((4-methoxybenzyl)amino)-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate 6 as a yellow solid (5.00 g, 12.6 mmol, 50.0% yield). LC-MS (ESI) m / z = 395.1 [M+H] +

[0096] Step 5: Methyl 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate (6) A solution of methyl 7-fluoro-4-((4-methoxybenzyl)amino)-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate 6 (2.00 g, 5.07 mmol, 1 equiv) in TFA (20.0 mL) is stirred at 75 °C for 16 h. Complete consumption of the starting material is indicated by LCMS and the required mass is detected. The reaction mixture is concentrated under reduced pressure to afford methyl 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate 7 as a yellow solid (1.39 g, crude product). LC-MS (ESI) m / z = 275.0 [M+H] +

[0097] Step 6: 4-Amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A7) A solution of methyl 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylate 7 (1.39 g, 5.07 mmol, 1 equiv) in EtOH (15 mL) / H2O (5.00 mL) is added to LiOH (638 mg, 15.2 mmol, 3 equiv). The mixture is stirred at 25 °C for 12 h. LCMS indicates complete consumption of the starting material and one major peak with the required mass is detected. The reaction mixture is diluted with H2O (20.0 mL) and extracted with (DCM 10.0 mL × 2) to remove impurities. The aqueous layer is adjusted to pH = 6 with 2M HCl, the precipitate is collected and dried under reduced pressure to afford 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A7) as a white solid (1.30 g, 5.00 mmol, 98.5% yield). 1 H NMR (400 MHz, DMSO-d6) δ 13.15 (br s, 1H), 9.12 (s, 1H), 8.56 (br d, J = 6.4 Hz, 1H), 7.35 (br s, 2H), 7.13 (br d, J = 12.1 Hz, 1H), 2.62 (s, 3H). LC-MS (ESI) m / z = 261.0 [M+H] +

[0098] General method: Synthesis of Intermediate B1 Synthetic route:

Chemical Structure

[0099] Step 2: Pyrazolo[1,5-a]pyridine-2-carboxaldehyde (3) To a solution of N-methoxy-N-methylpyrazolo[1,5-a]pyridine-2-carboxamide (1.5 g, 7.31 mmol) in THF (20 mL) at -60 °C is added LiAlH4 (439 mg, 10.96 mmol). The mixture is then stirred at -60 °C for 2 h. The reaction is quenched with water (40 mL) and filtered. The filtrate is concentrated under reduced pressure. The residue is purified by silica gel chromatography, eluting with a DCM solution of MeOH from 0% to 10% over 20 min to give pyrazolo[1,5-a]pyridine-2-carboxaldehyde as a yellow oil (450 mg, 42% yield). LC-MS: Rt = 1.071 min, (ESI) m / z. [M+H] + 147.1; C8H6N2O.

[0100] Step 3: N-(Pyrazolo[1,5-a]pyridin-2-ylmethyl)-1-(pyrimidin-2-yl)ethan-1-amine (Intermediate B1) To a solution of pyrazolo[1,5-a]pyridine-2-carboxaldehyde (100 mg, 0.68 mmol) in MeOH (5 mL) are added 1-(pyrimidin-2-yl)ethylamine (126 mg, 1.03 mmol) and NaBH3CN (86 mg, 1.37 mmol), and the reaction is carried out at 25 °C. Next, the mixture is stirred at 25 °C for 2 hours. The reaction is quenched with water (1 mL), and the solvent is concentrated under reduced pressure. The residue is purified by silica gel chromatography, eluting over 20 minutes from 0% to 5% DCM solution of MeOH to give N-(pyrazolo[1,5-a]pyridin-2-ylmethyl)-1-(pyrimidin-2-yl)ethylamine (40 mg, 23% yield) as a yellow oil. LC-MS: Rt = 0.521 min, (ESI) m / z. [M + H] + 254.2; C14H15N5。

[0101] General method: Synthesis of Intermediate B2 Synthetic route:

Chemical formula

[0102] Step 1: 2-Dichloromethyl-6-trifluoromethylimidazo[1,2-a]pyridine (3) A mixture of 5-(trifluoromethyl)pyridin-2-amine (1) (30 g, 185 mmol, 1 equiv), chlorobenzene (450 mL), and 1,1,3-trichloro-2-propanone (45 g, 277 mmol, 1.5 equiv) is reacted at 135 °C for 4 hours. The formation of the target product is detected by LCMS. The pH of the reaction solution is adjusted to about 8 with sodium carbonate, extracted with ethyl acetate (500 mL × 3), the organic phases are combined, dried over magnesium sulfate, the filtrate is filtered, concentrated to dryness under reduced pressure, and purified by column chromatography (silica, 15% ethyl acetate in petroleum ether) to give the yellow solid 2-dichloromethyl-6-trifluoromethylimidazo[1,2-a]pyridine (3) (30 g, 111 mmol, 60% yield). HnmR: ES19506-784-P1A, 11H NMR (400 MHz, DMSO-d6) δ 9.24 (s, 1H), 8.27 (s, 1H), 7.79 (d, J = 9.68 Hz, 1H), 7.65 (s, 1H), 7.56 (dd, J = 1.65, 9.57 Hz, 1H).

[0103] Step 2: 6-(Trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (4) 2-Dichloromethyl-6-trifluoromethylimidazo[1,2-a]pyridine (3) (30 g, 111 mmol, 1 equiv), water (600 mL), calcium carbonate (33 g, 334 mmol, 3 equiv) are heated to 100 °C and reacted for 2 h. The formation of the target product is detected by LCMS. Celite and ethyl acetate (600 mL) are added to the reaction solution, stirred at room temperature for 30 min, filtered, and extracted with ethyl acetate (600 mL × 2). The organic phases are combined, dried over magnesium sulfate, filtered, and the filtrate is concentrated to dryness under reduced pressure to obtain 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (4) (35 g) as a brown solid, which is used in the next step without further purification. HnmR: ES19506-789-P1A1, 1 1H NMR (400 MHz, CHLOROFORM-d) δ 10.09 - 10.29 (m, 1H), 8.59 (s, 1H), 8.27 (s, 1H), 7.82 (br d, J = 9.46 Hz, 1H), 7.44 (br d, J = 9.02 Hz, 1H).

[0104] Step 3: 1-Methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B2) A solution of 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (4) (100 mg, 467 μmol, 1 equiv) in DCM (2.00 mL) was added with KOAc (91.7 mg, 934 μmol, 2 equiv) and 1-methyl-1H-pyrazol-4-amine (45.4 mg, 467 μmol, 1 equiv) at -5 °C, and the reaction mixture was stirred at -5 °C for 1 h. Next, NaBH(OAc)3 (198 mg, 934 μmol, 2 equiv) was added and stirred at -5 °C for 3 h. LCMS (ET63219-45-P1A1) indicated complete consumption of Cpd.4 and showed several new peaks by LCMS. The reaction mixture was diluted with saturated aqueous Na2CO3 solution (3.00 mL) and extracted with dichloromethane (2.00 mL × 4). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (ethyl acetate / methanol = 8 / 1) to give 1-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B2) as a yellow solid (80.0 mg, 271 μmol, 58.0% yield). 1 H NMR (400 MHz, CDCl3) δ 8.47 (s, 1H), 7.69 (d, J = 9.5 Hz, 1H), 7.64 (s, 1H), 7.35 (d, J = 1.5, 9.5 Hz, 1H), 7.30 (s, 2H), 6.97 (s, 1H), 4.39 (s, 2H), 3.81 (s, 3H). LC-MS, [M+H] + 217.0.

[0105] General method: Synthesis of Intermediate B3 Synthetic route:

Chemical formula

[0106] Step 1: 1,3-Dimethyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B3) 6-(Trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (4) (10 g, 46 mmol, 1 equiv) and 1,3-dimethylpyrazol-4-amine (6 g, 56 mmol, 1.2 equiv) were dissolved in dichloromethane (150 mL), acetic acid (3 g, 56 mmol, 3 mL, 1.2 equiv) was added, and the reaction mixture was reacted at 25 °C for 1 h. Sodium triacetoxyborohydride (25 g, 117 mmol, 2.5 equiv) was added, and the reaction mixture was reacted at 25 °C for 3 h. The formation of the target product was detected by LCMS. The reaction mixture was quenched with 200 mL of sodium bicarbonate and extracted with ethyl acetate (150 mL × 2). The combined organic phases were washed with 400 mL of saturated brine, dried over magnesium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. Purification by column chromatography (silica, 35% ethyl acetate in petroleum ether:ethanol (3:1)) gave the brown solid 1,3-dimethyl-N-(6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)-1H-pyrazol-4-amine (Intermediate B3) (14 g, 45 mmol, 97% yield). HnmR: ES19506-795-P1A, 1 H NMR (400 MHz, DMSO-d6) δ 9.19 (s, 1H), 7.92 (s, 1H), 7.67 (d, J = 9.46 Hz, 1H), 7.41 (dd, J = 1.76, 9.46 Hz, 1H), 6.93 (s, 1H), 4.56 (br s, 1H), 4.17 (br d, J = 3.96 Hz, 2H), 3.52 - 3.61 (m, 3H), 2.04 (s, 3H). LCMS: ES19506-795-P1B, (ESI) m / z = 310.3 (M+1)+, RT = 0.64 min.

[0107] General method: Synthesis of Intermediate B5 Synthetic route:

Chemical formula

[0108] Step 1: Pyrazolo[1,5-a]pyridine-2-carboxylic acid (2) To a solution of pyrazolo[1,5-a]pyridine-2-carboxylic acid (10.0 g, 61.67 mmol, 1 equiv), HATU (28.14 g, 74.01 mmol, 1.2 equiv), and DIEA (31.88 g, 246.69 mmol, 4 equiv) in DCM (500 mL) is added N,O-dimethylhydroxylamine hydrochloride (12.03 g, 123.35 mmol, 2 equiv). The mixture is stirred at 25 °C for 16 h. LCMS (ET63565-18-P1A) indicates complete consumption of the starting material and detection of the product. The reaction mixture is concentrated under reduced pressure to give a residue. The residue is purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1 to 0 / 1) to afford the compound pyrazolo[1,5-a]pyridine-2-carboxylic acid (2) (12.1 g, 58.9 mmol, 95.6% yield) as a white solid. 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.49 (d, J = 7.1 Hz, 1H), 7.57 (d, J = 8.9 Hz, 1H), 7.15 (dd, J = 7.3, 8.4 Hz, 1H), 7.00 (s, 1H), 6.85 (t, J = 6.9 Hz, 1H), 3.79 (s, 3H), 3.49 (s, 3H), LC-MS, [MH] + 206.22.

[0109] Step 2: 1-(Pyrazolo[1,5-a]pyridin-2-yl)ethan-1-one (3) Under a nitrogen gas atmosphere, MeLi (11.70 mL, 1.2 equiv) is added to a solution of pyrazolo[1,5-a]pyridine-2-carboxylic acid (2) (2 g, 9.75 mmol, 1 equiv) in THF (20.0 mL) at -60 °C, and then the mixture is stirred at 25 °C for 16 h. LCMS (ET63565-13-P1A1) indicates complete consumption of the starting material and detection of the product. The residue is purified by column chromatography (petroleum ether / ethyl acetate = 50 / 1 to 3 / 1) to afford 1-(Pyrazolo[1,5-a]pyridin-2-yl)ethan-1-one (3) (277 mg, 1.73 mmol, 17.7% yield) as a white solid. LC-MS, [MH] + 161.1.

[0110] Step 3: N-Ethyl-1-(pyrazolo[1,5-a]pyridin-2-yl)ethan-1-amine (Intermediate B5) A mixture of 1-(pyrazolo[1,5-a]pyridin-2-yl)ethan-1-one (3) (0.1 g, 624.33 μmol, 1 eq) and ethylamine (112.58 mg, 2.50 mmol, 163.39 μL, 4 eq) in DCM (2 mL) was stirred at -60 °C for 30 minutes, then NaBH4 (264.64 mg, 1.25 mmol, 2 eq) was added to the mixture at 0 °C, and the mixture was stirred at 25 °C for 16 hours under a N2 atmosphere. Complete consumption of the starting material was indicated by LCMS (ET63565-9-P1A2), and the product was detected. The reaction mixture was quenched with H2O (5 mL) and diluted with DCM (5 mL × 3), and the combined organic layers were concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (DCM:MeOH = 10:1) to give N-Ethyl-1-(pyrazolo[1,5-a]pyridin-2-yl)ethan-1-amine (Intermediate B5) as a white solid (50 mg, 264 μmol, 42.3% yield). 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.39 (br d, J = 6.9 Hz, 1H), 7.46 (br d, J = 8.5 Hz, 1H), 7.12 - 7.03 (m, 1H), 6.70 (br t, J = 6.8 Hz, 1H), 6.44 (s, 1H), 4.13 (br d, J = 6.8 Hz, 1H), 3.32 - 3.21 (m, 1H), 2.77 - 2.56 (m, 2H), 1.53 (d, J = 6.6 Hz, 4H), 1.17 - 1.14 (m, 3H): LC-MS, [MH] + 190.1.

[0111] General method: Synthesis of Intermediate B6 Synthetic route:

Chemical formula

[0112] Step 1: 2-(Dichloromethyl)-6-(trifluoromethyl)-1H-benzo[d]imidazole (3) A mixture of 4-(trifluoromethyl)benzene-1,2-diamine (5 g, 28.3 mmol, 1 eq) and dichloroacetic acid (7.32 g, 56.7 mmol, 4.66 mL, 2 eq) in HCl (125 mL) (4 M) is stirred at 100 °C for 10 minutes. For 16 hours. Consumption of Cpd.1 is shown by LCMS (ET60224-68-P1A) and the required mass is detected. The reaction mixture is filtered and the filter cake is washed with water. The combined filtrate is extracted with DCM (20 ml×3). The combined organic layers are washed with brine (100 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to give a residue. The residue is purified by column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 5 / 1) to give the yellow oily compound 2-(dichloromethyl)-6-(trifluoromethyl)-1H-benzo[d]imidazole (3) (4.4 g, 16.3 mmol, 57.6% yield). 1 1H NMR (400 MHz, CHLOROFORM-d) δ 8.01 (s, 1H), 7.79 (d, J = 8.6 Hz, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.26 (s, 1H); LCMS: [M+H] + 268.9

[0113] Step 2: 6-(Trifluoromethyl)-1H-benzo[d]imidazole-2-carboxaldehyde (4) Calcium carbonate (1.12 g, 11.1 mmol, 3 eq) is added to a suspension of 2-(dichloromethyl)-6-(trifluoromethyl)-1H-benzo[d]imidazole (3) (1 g, 3.72 mmol, 1 eq) in H2O (20 mL). The mixture is stirred at 100 °C for 8 hours. Consumption of Cpd.3 is shown by LCMS (ET60224-74-P1B) and the required mass is detected. The reaction mixture is diluted with H2O (50 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layers are concentrated under reduced pressure to give 6-(trifluoromethyl)-1H-benzo[d]imidazole-2-carboxaldehyde (4) as a white solid (310 mg, 1.45 mmol, 38.9% yield). 11H NMR (400 MHz, DMSO-d6) δ 14.28 - 13.59 (m, 1H), 10.02 (s, 1H), 8.40 - 8.30 (m, 2H), 8.23 - 8.05 (m, 3H), 8.00 (dd, J = 5.4, 8.5 Hz, 1H), 7.89 (br s, 1H), 7.83 - 7.61 (m, 3H), 7.34 (br d, J = 7.9 Hz, 1H), 7.27 (d, J = 7.6 Hz, 1H); LCMS: [M+H] + 215.2。

[0114] Step 3: 2-Methyl-N-((6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)methyl)propan-1-amine (Intermediate B6) At 25 °C, KOAc (170 mg, 1.74 mmol, 1.2 eq) is added to a solution of 6-(trifluoromethyl)-1H-benzo[d]imidazole-2-carboxaldehyde (4) (0.31 g, 1.45 mmol, 1 eq) and 2-methylpropan-1-amine (105 mg, 1.45 mmol, 143 μL, 1 eq) in DCM (6.2 mL). The mixture is stirred at 25 °C for 30 minutes, then NaBH(OAc)3 (398 mg, 1.88 mmol, 1.3 eq) is added to the above mixture at 25 °C, and the mixture is stirred at 25 °C for 15.5 hours. Consumption of Cpd.4 is indicated by LCMS (ET60224 - 77 - P1A), and the required mass is detected. The reaction mixture is diluted with H2O (10 mL) and extracted with DCM (2 mL × 3). The combined organic layers are washed with brine (10 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue is purified by preparative TLC (petroleum ether / ethyl acetate = 0 / 1) to obtain 2-methyl-N-((6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)methyl)propan-1-amine (Intermediate B6) as a colorless oil (100 mg, 368 μmol, 25.4% yield, 100% purity). 11H NMR (400 MHz, CHLOROFORM-d) δ 7.80 (s, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 8.5 Hz, 1H), 4.07 (s, 2H), 2.45 (d, J = 6.8 Hz, 2H), 1.82 - 1.68 (m, 1H), 0.89 (d, J = 6.6 Hz, 6H); LCMS: [M+H] + 272.0

[0115] General method: Synthesis of Intermediate B7

Chemical Structure

[0116] Step: 2-Methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)propan-1-amine (Intermediate B7) To a solution of 2-methylpropan-1-amine (68.3 mg, 933 μmol, 92.8 μL, 1 equiv) and 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (0.2 g, 933 μmol, 1 equiv) in DCM (4 mL) was added KOAc (109 mg, 1.12 mmol, 1.2 equiv). The mixture was stirred at 25 °C for 30 min at 25 °C, then NaBH(OAc)3 (257 mg, 1.21 mmol, 1.3 equiv) was added to the above mixture at 25 °C, and the mixture was stirred at 25 °C for 15.5 h. Consumption of Cpd.4 was indicated by LCMS (ET60224-75-P1A), and the required mass was detected. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (2 mL × 3) to remove impurities. The aqueous layer was basified to pH = 8 with saturated Na2CO3 and then extracted with DCM (10 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to afford the intermediate 2-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)propan-1-amine (Intermediate B7) as a colorless oil (108 mg, 398 μmol, 42.6% yield, 100% purity). 11H NMR (400 MHz, CHLOROFORM-d) δ 8.49 (s, 1H), 7.70 - 7.62 (m, 2H), 7.33 (br d, J = 9.2 Hz, 1H), 4.01 (s, 2H), 2.54 (d, J = 6.7 Hz, 2H), 1.84 (quind, J = 6.6, 13.3 Hz, 1H), 0.97 (d, J = 6.6 Hz, 6H), LC-MS, [MH] + 272.0

[0117] Synthesis of Example 4

Chemical Structure

[0118] Step 7: (E)-4-(((Dimethylamino)methylene)amino)-3-methylpyrazolo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A6-1) HCl / dioxane (4 M) is added to a solution of 4-amino-3-methylpyrazolo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A6) (50.0 mg, 206 μmol, 1 equiv) in DCM (2 mL). The mixture is stirred at 25 °C for 30 minutes. The reaction mixture is then concentrated, evaporated with hexane (10 mL × 3), and cooled to 0 °C. Oxalyl chloride (157 mg, 1.24 mmol, 108 μL, 6 equiv) and DMF (3.02 mg, 41.3 μmol, 3.18 μL, 0.2 equiv) are added dropwise at 0 °C. The mixture is stirred at 0 °C for 2 hours. A sample is taken out from MeOH. LC-MS shows that the methyl ester of the required mass is detected. The mixture is concentrated under reduced pressure to obtain a residue. The residue is triturated with n-hexane (5 mL) and then concentrated under reduced pressure to obtain (E)-4-(((dimethylamino)methylene)amino)-3-methylpyrazolo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A6-1) (50.0 mg, 158 μmol, 76.7% yield) as a yellow solid. LC-MS, [MH] + 243.1

[0119] Step 8: (E)-4-(((Dimethylamino)methylene)amino)-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Intermediate A6-2) (E)-4-(((Dimethylamino)methylene)amino)-3-methylpyrazolo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A6-1) (50.0 mg, 158 μmol, 1 eq) and DIEA (81.9 mg, 633 μmol, 110 μL, 4 eq) in THF (2 mL) solution, Int.6 (44.7 mg, 158 μmol, 1 eq) is added at 0 °C. The mixture is stirred at 0 °C for 2 h. Complete consumption of the starting material is indicated by LC-MS and the required mass is detected. The mixture is concentrated under reduced pressure to give (E)-4-(((Dimethylamino)methylene)amino)-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Intermediate A6-2) (61.0 mg, 109 μmol, 68.6% yield) as a yellow solid. LC-MS, [MH] + 562.2。

[0120] Step 9: 4-Amino-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Example 4) A solution of (E)-4-(((dimethylamino)methylene)amino)-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Intermediate A6-2) (60.0 mg, 107 μmol, 1 eq) in NH3 / MeOH (2 mL) is stirred at 70 °C for 2 h. Complete consumption of the starting material is indicated by LC-MS and the required mass is detected. The reaction mixture is concentrated under reduced pressure to give a residue. The residue is purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150×40 mm×10 μm, mobile phase: [water (NH4HCO3)-ACN], B%: 25% - 65%, 8 min, UV220&254 nm) to afford 4-amino-3-methyl-N-(1-(pyrimidin-2-yl)ethyl)-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)pyrazolo[1,5-a]quinoxaline-8-carboxamide (Example 4) as a white solid (19.2 mg, 37.9 μmol, 35.48% yield, 100% purity). 1 H NMR (400 MHz, DMSO-d6) δ 8.84 (br s, 1H) 8.79 (br d, J = 4.63 Hz, 2H) 8.28 (br s, 1H) 8.13 (br d, J = 6.75 Hz, 1H) 7.80 - 8.01 (m, 1H) 7.49 - 7.72 (m, 3H) 7.29 - 7.47 (m, 1H) 6.99 (br s, 2H) 5.43 (br s, 1H) 4.92 (br d, J = 17.01 Hz, 1H) 4.52 (br d, J = 16.51 Hz, 1H) 2.50 (br s, 3H) 1.62 (br d, J = 7.00 Hz, 3H). LC-MS, [MH] + 507.2。

[0121] Synthesis of Example 15

Chemical Structure

[0122] Step 1: 4-(2,4-Dimethoxybenzyl)amino)-N-ethyl-1,3-dimethyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (3) 4-((2,4-Dimethoxybenzyl)amino)-1,3-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid (h) (100 mg, 246.0 μmol, 1 equiv) was dissolved in N,N-dimethylformamide (2.5 mL), and N-(5-(trifluoromethyl)pyridin-2-ylmethyl)ethylamine (60.3 mg, 295.2 μmol, 1.2 equiv), N,N,N,N-tetramethylchloroformamidinium hexafluorophosphate (138 mg, 492.1 μmol, 2 equiv), and N-methylimidazole (101 mg, 1.23 mmol, 98.1 μL, 5 equiv) were added. The reaction mixture was reacted at 50 °C for 16 h. LCMS indicated complete consumption of the starting material and formation of the desired product. The reaction mixture was concentrated to dryness under reduced pressure and purified by column chromatography (ISCOR, 4 g SepaFlashR silica gel column, 0 - 2.3% methanol / dichloromethane system, flow rate 20 mL / min) to obtain 4-(2,4-dimethoxybenzyl)amino)-N-ethyl-1,3-dimethyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (3) as a yellow oil (150 mg, 233.45 μmol, 94.88% yield, 92.23% purity). LCMS: ES19974-409-P1B1, (ESI) m / z = 593.2 [M+1] + , RT = 0.82 min, purity: 92.23%

[0123] Step 2: 4-Amino-N-ethyl-1,3-dimethyl-N-((5-trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 15) 4-(2,4-Dimethoxybenzyl)amino)-N-ethyl-1,3-dimethyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (3) (140 mg, 236.2 μmol, 1 equiv) was dissolved in dichloromethane (0.5 mL), trifluoroacetic acid (2.5 mL) was added, and the reaction mixture was reacted at 50 °C for 16 h. Complete consumption of the starting material was detected by LC-MS, and the target product was formed. The reaction mixture was concentrated to dryness under reduced pressure, 10% sodium carbonate (10 mL) was added, and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic phases were washed with saturated brine (10 mL), dried over magnesium sulfate, filtered, concentrated to dryness, and purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 150 × 30 mm × 5 μm, mobile phase: [water (ammonium hydroxide)-acetonitrile], B% gradient: 30% - 50%, 9 min) to obtain white solid 4-amino-N-ethyl-1,3-dimethyl-N-((5-trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 15) (30 mg, 67.81 μmol, 28.70% yield). LCMS: ES19974-414-P1B1, (ESI) m / z = 443.1 [M+1] + , RT = 2.623 min, purity: 100.0%. 1H NMR: ES19974-414-P1A, 1 1H NMR (400 MHz, DMSO-d6) Shift = 8.97 (br s, 1H), 8.20 (br d, J = 6.9 Hz, 1H), 7.97 (br s, 1H), 7.61 (br d, J = 8.1 Hz, 1H), 7.39 (br s, 2H), 6.72 (br s, 2H), 4.82 (br s, 2H), 3.45 (q, J = 6.9 Hz, 2H), 3.10 - 2.60 (m, 3H), 2.56 (br s, 3H), 1.14 (t, J = 7.0 Hz, 3H)

[0124] Synthesis of Example 86

Chemical Structure

[0125] Step 1: 7-Chloro-4-[(2,4-dimethoxyphenyl)methylamino]-1-methyl-N-(1-methylpyrazol-4-yl)-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (3) 7-Chloro-4-((2,4-dimethoxybenzyl)amino)-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A3) (145 mg, 339 μmol, 1 equiv) was added to a solution of 1-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-4-amine (Intermediate B2) (100 mg, 339 μmol, 1 equiv) in acetonitrile (2 mL), and after dissolution, N,N,N,N-tetramethylchloroformamidinium hexafluorophosphate (285 mg, 1 mmol, 3 equiv) and N-methylimidazole (139 mg, 1.7 mmol, 135 μL, 5 equiv) were sequentially added. The reaction mixture was stirred at 50 °C for 16 h. Completion of the reaction of the starting materials was detected by LC-MS, and the target product was formed. After filtering the reaction solution, it was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (silica, 10% methanol in dichloromethane) to obtain 7-chloro-4-[(2,4-dimethoxyphenyl)methylamino]-1-methyl-N-(1-methylpyrazol-4-yl)-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (3) as a yellow oil (130 mg, 185 μmol, 54.5% yield).

[0126] Step 2: 4-Amino-7-chloro-1-methyl-N-(1-methyl-1H-pyrazol-4-yl)-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl])methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 86) 7-Chloro-4-[(2,4-dimethoxyphenyl)methylamino]-1-methyl-N-(1-methyl-1H-pyrazol-4-yl)-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (3) (120 mg, 170 μmol, 1 equiv) was added to a mixed solution of trifluoroacetic acid (0.4 mL) and dichloromethane (1 mL) and dissolved. The reaction mixture was stirred at 50 °C for 16 h. Complete reaction of the starting material was indicated by LC-MS, and the desired product was formed. The reaction mixture was concentrated to dryness under reduced pressure. It was purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water and acetonitrile with decreasing polarity as the eluent) to give white 4-amino-7-chloro-1-methyl-N-(1-methyl-1H-pyrazol-4-yl)-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl])methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 86) (30 mg, 53 μmol, 31% yield, 98.% purity). 1H NMR: 1 1H NMR (400 MHz, DMSO-d6) δ 9.15 - 9.31 (m, 1H), 8.33 (s, 1H), 8.07 - 8.16 (m, 1H), 7.92 - 8.03 (m, 1H), 7.76 - 7.86 (m, 1H), 7.58 - 7.75 (m, 2H), 7.41 - 7.55 (m, 3H), 7.18 - 7.33 (m, 1H), 4.71 - 5.17 (m, 2H), 3.53 - 3.83 (m, 3H), 2.80 - 2.97 (m, 3H). LCMS: (ESI) m / z = 554.2 (M+1)+, RT = 1.544 min, purity of 98.7%.

[0127] Synthesis of Example 143 Synthesis method:

Chemical Structure

[0128] Step 1: 2-Methyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) 5-(Trifluoromethyl)pyridinecarboxaldehyde (2) (300 mg, 1.71 mmol, 1.0 equiv), 2-methylpyridin-3-amine (1) (216 mg, 2.00 mmol, 1.17 equiv), and acetic acid (123 mg, 2.05 mmol, 117.25 μL, 1.2 equiv) were dissolved in dichloromethane (10 mL), stirred at room temperature for 4 h, sodium triacetoxyborohydride (1.09 g, 5.15 mmol, 3.01 equiv) was added, and the mixture was stirred at room temperature for 12 h. Complete consumption of the starting materials was detected by LCMS, and the product was formed. Water and dichloromethane were added to the reaction mixture for dilution, the pH was adjusted to approximately 12 with 2 mol aqueous potassium carbonate, and the layers were separated. The aqueous phase was extracted with dichloromethane (10 mL × 3), the organic phases were combined, dried over sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography (ISCOR, 12 g SepaFlashR silica gel column, 0 - 25% ethyl acetate / petroleum ether system, flow rate, 30 mL / min) to obtain 2-methyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) as a yellow syrup (205 mg, 44.7% yield). LCMSES15882-1182-P1A: (ESI) m / z = 268.1 [M+1] + , RT = 0.65 min

[0129] Step 2: 4-Amino-7-fluoro-1-methyl-N-(2-methylpyridin-3-yl)-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 143) 2-Methyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) (60 mg, 224.51 μmol, 1.0 equiv), 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (60 mg, 230.57 μmol, 1.03 equiv), and N-ethyl-N-isopropyl-2-propanamine (119 mg, 925.11 μmol, 161.13 μL, 4.12 equiv) were dissolved in 1-methylpyrrolidin-2-one (1.0 mL), 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (DMC) (57.82 mg, 342.02 μmol, 1.52 equiv) was added, and the reaction mixture was reacted at 80 °C for 16 h. The formation of the target product was detected by LCMS. The reaction mixture was purified by reverse-phase preparative liquid chromatography (formic acid condition, Boston Prime C18 column, 150×30 mm×5 μm, mobile phase: [water (formic acid)-acetonitrile], gradient: 15% - 35% B, 10 min) to obtain a yellow solid, 4-amino-7-fluoro-1-methyl-N-(2-methylpyridin-3-yl)-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 143) (8 mg, 6.29% yield). LCMSES15882-1205-P1B1: (ESI) m / z = 510.1 [M+1] + , RT = 0.996 min. 1 H NMR (400 MHz, DMSO-d6) Shift 8.84 - 8.98 (m, 1H), 8.17 - 8.27 (m, 2H), 7.72 - 7.87 (m, 4H), 7.50 (s, 2H), 7.13 (dd, J = 4.64, 7.91 Hz, 1H), 7.01 (d, J = 11.29 Hz, 1H), 5.44 (d, J = 15.81 Hz, 1H), 5.01 (d, J = 15.81 Hz, 1H), 2.82 (s, 3H), 2.37 (s, 3H) 19FnmR (376.5 MHz, DMSO-d6) Shift -60.71, -60.85, -116.51, -119.18

[0130] Synthesis of Example 145 Synthesis method: [Chemical]

[0131] Step 1: 2-Methoxy-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) 5-(Trifluoromethyl)pyridinecarboxaldehyde (2) (300 mg, 1.71 mmol, 1.0 equiv), 2-methoxypyridin-3-amine (1) (248 mg, 2.00 mmol, 1.17 equiv), and acetic acid (123 mg, 2.05 mmol, 1.2 equiv) are dissolved in dichloromethane (10 mL) and stirred at room temperature for 4 h. Sodium triacetoxyborohydride (1.09 g, 5.15 mmol, 3.01 equiv) is added and stirred at room temperature for 12 h. Complete consumption of the starting materials is detected by LCMS and the product is formed. Water and dichloromethane are added to the reaction solution for dilution, and the pH is adjusted to about 12 with 2 mol aqueous potassium carbonate solution. The layers are separated, and the aqueous phase is extracted with dichloromethane (10 mL × 3). The organic phases are combined, dried over sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography (ISCOR, 12 g SepaFlashR silica gel column, 0 - 15% ethyl acetate / petroleum ether system, flow rate, 30 mL / min) to obtain 2-methoxy-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) as a yellow syrup (430 mg, 1.52 mmol, 88.61% yield). LCMSES15882-1189-P1A: (ESI) m / z = 284.0 [M+1] + , RT = 0.82 min

[0132] Step 2: 4-Amino-7-fluoro-N-(2-methoxypyridin-3-yl)-1-methyl-N-(5-(trifluoromethyl)pyridin-2-ylmethyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 145) 2-Methoxy-N-(5-trifluoromethyl)pyridin-2-ylmethyl)pyridin-3-amine (3) (64 mg, 226 μmol, 1.0 equiv), 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (61 mg, 234 μmol, 1.04 equiv), and N-ethyl-N-isopropyl-2-propylamine (119 mg, 919 μmol, 4.07 equiv) were dissolved in 1-methylpyrrolidin-2-one (1.0 mL), 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (DMC) (58 mg, 344 μmol, 1.52 equiv) was added, and the reaction mixture was reacted at 80 °C for 16 h. The formation of the target product was detected by LCMS. The reaction mixture was purified by reverse-phase preparative liquid chromatography (formic acid condition, Boston Prime C18 column, 150×30 mm×5 μm, mobile phase: [water (formic acid)-acetonitrile], gradient: 23% - 43% B, 10 min) to obtain the yellow solid 4-amino-7-fluoro-N-(2-methoxypyridin-3-yl)-1-methyl-N-(5-trifluoromethyl)pyridin-2-methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 145) (12 mg, 8.79% yield, 87% purity). LCMSES15882-1206-P1B1: (ESI) m / z = 526.1 [M+1] + , RT = 1.140 min 1 H NMR (400 MHz, DMSO-d6) Shift 8.91 (s, 1H), 8.23 - 8.30 (m, 1H), 8.20 (s, 0.2H, HCOOH), 7.91 - 7.95 (m, 1H), 7.86 (d, J = 6.53 Hz, 1H), 7.77 - 7.83 (m, 2H), 7.72 - 7.76 (m, 1H), 7.50 (br s, 2H), 7.03 (d, J = 11.29 Hz, 1H), 6.89 (dd, J = 5.02, 7.53 Hz, 1H), 5.34 (d, J = 16.06 Hz, 1H), 5.04 (d, J = 15.81 Hz, 1H), 3.74 (s, 3H), 2.80 (s, 3H) 19FnmR (376.5 MHz, DMSO-d6) Shift -60.72, -117.24

[0133] Synthesis of Example 188 Synthesis method:

Chemical formula

[0134] Step 1: 1,3-Dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-pyrazole-4-amine (2) Dissolve 5-(trifluoromethyl)pyridinecarboxaldehyde (1) (300 mg, 1.71 mmol, 1 equivalent) in dichloromethane (8 mL), add 1,3-dimethyl-1H-pyrazole-4-amine (247 mg, 2.22 mmol, 1.3 equivalents) and glacial acetic acid (134 mg, 2.23 mmol, 127.74 μL, 1.3 equivalents), and stir the reaction mixture at 15 - 20 °C for 4 hours. Add sodium borohydride acetate (1.09 g, 5.15 mmol, 3.01 equivalents), and stir at 15 - 20 °C for 12 hours. The completion of the reaction of the raw materials is detected by LC-MS, and the target product is formed. Add 10 mL of water and 10 mL of dichloromethane to the reaction solution, basify the pH to 12 with 2 mol / L aqueous potassium carbonate solution, separate the layers, extract with dichloromethane (10 mL × 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate to dryness under reduced pressure, and purify by column chromatography (12 g silica gel column, eluent 0 - 40% (ethyl acetate:ethanol = 3:1) / petroleum ether, flow rate @ 30 mL / min), and concentrate under reduced pressure to obtain a yellow solid 1,3-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-pyrazole-4-amine (3) (311 mg, 1.15 mmol, 67.17% yield). LCMSES15882-1166-P1A: (ESI) m / z = 228.2 [M + 1] + , RT = 0.601 min

[0135] Step 2: 4-Amino-N-(1,3-dimethyl-1H-pyrazol-4-yl)-1,7-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 188) The reaction solution: 4-amino-1,7-dimethylimidazo[1,5-a]quinoxaline-8-carboxylic acid (c) (42 mg, 163.90 μmol, 1 equivalent), 1,3-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-pyrazol-4-amine (b) (45.18 mg, 167.17 μmol, 1.02 equivalents), and N,N-diisopropylethylamine (84.73 mg, 655.59 μmol, 114.19 μL, 4 equivalents) were dissolved in 1-methyl-2-pyrrolidone (1 mL), and 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (41.56 mg, 245.84 μmol, 1.5 equivalents) was added. The reaction mixture was stirred at 50 °C for 3 hours. The complete reaction of the starting materials was detected by LC-MS, and the target product was formed. Purification was carried out by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water (containing 0.05% formic acid) and acetonitrile (20%-40%) with decreasing polarity as the eluent) to obtain a yellow solid. The crude product was purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water (containing 0.05% formic acid) and acetonitrile (18%-38%) with decreasing polarity as the eluent) to obtain the white solid 4-amino-N-(1,3-dimethyl-1H-pyrazol-4-yl)-1,7-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 188) (10 mg, 19.67 μmol, 12.00% yield). LCMS: ES13685-1063-P1D, (ESI) m / z = 509.1 [M+1] + , RT = 1.066 min NMR: ES13685-1063-P1A, 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.25 (br d, J = 8.60 Hz, 1H), 8.15 (s, 1H), 7.75 (s, 2H), 7.56 - 7.72 (m, 2H), 7.23 (br d, J = 4.84 Hz, 3H), 5.14 (s, 2H), 3.51 - 3.70 (m, 3H), 2.79 (s, 3H), 2.38 - 2.44 (m, 3H), 1.83 (s, 3H), 19F nmR (376 MHz, DMSO-d6) δ -60.68 (br s, 3F)

[0136] Synthesis of Example 210 Synthesis method:

Chemical formula

[0137] Step 1: 1,4-Dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-imidazole-2-amine (2) 2-Bromo-1,4-dimethyl-1H-imidazole (210 mg, 1.20 mmol, 1 equiv) was dissolved in tetrahydrofuran (12 mL), and (5-(trifluoromethyl)pyridin-2-yl)methylamine (1) (211 mg, 1.20 mmol, 1 equiv), sodium trimethylsilanide (267 mg, 1.44 mmol, 1.2 equiv) and bromo[bis(cyclohexyl)[3-(1,1-dimethylethoxy)-6-methoxy-2′,6′-bis(1-methylethyl)[1,1′-biphenyl]-2-yl-κC1′]phosphine-κP][4-[[2-(trimethylsilyl)ethoxy]carbonyl]phenyl]-,(SP-4-2)-palladium (GPhos-Pd-G6) (50 mg, 59.99 μmol, 0.05 equiv) were added under nitrogen gas protection, and the reaction mixture was stirred at 80 °C for 16 h. The completion of the reaction of the raw materials was detected by LC-MS, and the target product was formed. After cooling to room temperature, 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous magnesium sulfate, the filtrate was filtered, concentrated to dryness under reduced pressure, and purified by column chromatography (4 g + 4 g silica gel column, eluent 0 - 40% ethyl acetate / petroleum ether, flow rate 20 mL / min) to obtain 1,4-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-imidazole-2-amine (2) as a yellow liquid (50 mg, 185.01 μmol, 15.42% yield). LCMS:ES13685-1155-P1C, (ESI) m / z = 271.1 [M+1] + , RT = 0.926 min

[0138] Step 2: 4-Amino-N-(1,4-dimethyl-1H-imidazol-2-yl)-7-fluoro-1-methyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 210) The reaction solution of 4-amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (40 mg, 153.71 μmol, 1 equivalent), 1,4-dimethyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)-1H-imidazole-2-amine (2) (41.54 mg, 153.71 μmol, 1 equivalent) and N,N-diisopropylethylamine (79.46 mg, 614.86 μmol, 107.09 μL, 4 equivalents) was dissolved in 1-methyl-2-pyrrolidone (1 mL), and 2-chloro-1,3-dimethyl-4,5-dihydroimidazole-1-chloride (38.98 mg, 230.5 μmol, 1.5 equivalents) was added. The reaction mixture was stirred at 60 °C for 16 h. A small amount of the remaining starting material was detected by LC-MS, and the target product was formed. Purification was carried out by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water (containing 0.05% formic acid) and acetonitrile (10% - 30%) with decreasing polarity as the eluent) to obtain a yellow solid. The crude product was purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water (containing 0.05% ammonium hydroxide) and acetonitrile (28% - 48%) with decreasing polarity as the eluent) to obtain the white solid 4-amino-N-(1,4-dimethyl-1H-imidazol-2-yl)-7-fluoro-1-methyl-N-((5-(trifluoromethyl)pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 210) (3 mg, 5.56 μmol, 3.62% yield). LCMS: ES13685-1158-P1C1, (ESI) m / z = 513.3 [M+1] + , RT = 0.759 min NMR: ES13685-1158-P1A, 11H NMR (400 MHz, DMSO-d6) δ 8.91 (br s, 1H), 8.23 (br d, J = 8.13 Hz, 1H), 7.97 (br d, J = 6.75 Hz, 1H), 7.89 (br s, 1H), 7.81 (s, 1H), 7.34 (br s, 2H), 7.13 (br d, J = 11.26 Hz, 1H), 6.50 (s, 1H), 5.17 (br s, 2H), 3.23 - 3.28 (m, 3H), 2.87 (s, 3H), 1.96 (br s, 3H). NMR: ES13685-1158-P1A, 19F nmR (376 MHz, DMSO-d6) δ -60.85 (s, 3F), -118.24 (br s, 1F)

[0139] Synthesis of Example 213 Synthesis method:

Chem.

[0140] Step 1: 1-Methyl-N-(1-(trifluoromethyl)pyrazol-4-ylmethyl)-1H-pyrazol-5-amine (3) 1-Methylpyrazol-5-amine (133 mg, 1.37 mmol, 1.5 eq) and 1-(trifluoromethyl)pyrazole-4-carboxaldehyde (1) (150 mg, 914.16 μmol, 1 eq) were dissolved in methanol (5 mL), acetic acid (164 mg, 2.74 mmol, 156.99 μL, 3 eq) was added, and the reaction mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (172.34 mg, 2.74 mmol, 3 eq) was added and the mixture was stirred at room temperature for 15.5 hours. Completion of the reaction was detected by LCMS and the desired product was formed. The mixture was concentrated to dryness under reduced pressure, sodium bicarbonate (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and purified by column chromatography (ISCOR, 12 g SepaFlashR silica gel column, 0 - 60% ethyl acetate:ethanol (3:1) / petroleum ether system, flow rate, 35 mL / min) to obtain the colorless oil 1-methyl-N-(1-(trifluoromethyl)pyrazol-4-ylmethyl)-1H-pyrazol-5-amine (3) (220 mg, 897.21 μmol, 86.86% yield). LCMS: ES13683-1961-P1C, (ESI) m / z = 246.1 (M+1)+, RT = 0.786 min

[0141] Step 2: 4-Amino-7-fluoro-1-methyl-1-methylpyrazol-5-yl)-N-(1-(trifluoromethyl)-1H-pyrazol-4-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 213) 4-Amino-7-fluoro-1-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (63.6 mg, 244.69 μmol, 1 eq) and 1-methyl-N-(1-(trifluoromethyl)pyrazol-4-ylmethyl)-1H-pyrazol-5-amine (3) (60 mg, 244.69 μmol, 1 eq) were dissolved in 1-methylpyrrolidin-2-one (1.5 mL), and N-ethyl-N-isopropyl-2-propylamine (158 mg, 1.22 mmol, 213.11 μL, 5 eq) and 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (DMC) (62.0 mg, 367.04 μmol, 1.5 eq) were added. The reaction mixture was reacted at 50 °C for 16 h. Complete consumption of the starting materials was detected by LCMS, and the target product was formed. The reaction mixture was diluted with acetonitrile (1 mL) and water (1 mL), filtered, and the filtrate was purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column 150×30 mm×5 μm, mobile phase: [water (formic acid)-acetonitrile], gradient: 18% - 38%, 10 min) to obtain the white solid 4-amino-7-fluoro-1-methyl-1-(methylpyrazol-5-yl)-N-(1-(trifluoromethyl)-1H-pyrazol-4-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 213) (50 mg, 100.53 μmol, 41.08% yield, 98% purity). LCMS: ES13683-1966-P1C, (ESI) m / z = 488.3 [M+1] + , RT = 0.767 min HNMR: ES13683-1966-P1A, 1 1H NMR (400 MHz, DMSO-d6) Shift 8.39 (s, 1H), 8.17 (s, 0.2H), 7.84 - 8.07 (m, 2H), 7.80 (s, 1H), 7.52 (s, 2H), 7.23 (br s, 1H), 7.06 (br d, J = 11.01 Hz, 1H), 6.06 (br s, 1H), 4.49 - 5.28 (m, 2H), 3.48 - 3.74 (m, 3H), 2.87 (s, 3H). 19F NMR: 19FnmR (376 MHz, DMSO-d6) Shift -59.21 (s, 3F), -118.10 (br s, 1F)

[0142] Synthesis of Example 220 Synthesis method: [Chemical formula]

[0143] Step 1: 4-Fluoro-2-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-3-amine (3) Dissolve 6-(trifluoromethyl)imidazo[1,2-a]pyridine-2-carboxaldehyde (1) (700 mg, 3.27 mmol, 1 equivalent) in methanol (14 mL), add 4-fluoro-1-methyl-1H-pyrazol-5-amine (2) (402.61 mg, 3.50 mmol, 1.07 equivalents) and glacial acetic acid (255.18 mg, 4.25 mmol, 243.26 μL, 1.3 equivalents), and stir the reaction mixture at 25 °C for 1 hour. Add sodium cyanoborohydride (616.24 mg, 9.81 mmol, 3 equivalents), and stir at 25 °C for 15 hours. The completion of the reaction of the raw materials is detected by LC-MS, and the target product is formed. Concentrate the reaction solution, spin-dry it, add 14 mL of 2 mol / L aqueous sodium carbonate solution and 14 mL of ethyl acetate, separate the layers, extract with ethyl acetate (14 mL × 3), dry the organic phase over anhydrous magnesium sulfate, filter, concentrate the filtrate under reduced pressure to dryness, and purify by column chromatography (12 g + 4 g silica gel column, eluent 0 - 30% ethyl acetate:ethanol 3:1 / petroleum ether, flow rate 30 mL / min), and concentrate under reduced pressure to obtain a pale yellow solid, 4-fluoro-2-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-3-amine (3) (970 mg, 3.10 mmol, 94.73% yield). LCMS: ES13685-1213-P1A, (ESI) m / z = 314.0 [M+1] + , RT = 1.537 min. NMR: ES13685-1213-R2A, 11H NMR (400 MHz, CHLOROFORM-d) δ 7.11 (d, J = 4.38 Hz, 1H), 3.57 (s, 3H), 3.24 (br s, 2H). 1H NMR: ES13685-1213-R2A, 19F nmR (376 MHz, CHLOROFORM-d) δ -185.48 (s, 1F)

[0144] Step 2: 4-Amino-7-fluoro-N-(4-fluoro-1-methyl-1H-pyrazol-5-yl)-1-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin)-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 220) Reaction solution: 4-amino-7-fluoro-methyl-imidazo[1,5-a]quinoxaline-8-carboxylic acid (500 mg, 1.92 mmol, 1 equivalent), 4-fluoro-2-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]pyrazol-3-amine (3) (601.89 mg, 1.92 mmol, 1 equivalent), and N,N-diisopropylethylamine (993.30 mg, 7.69 mmol, 1.34 mL, 4 equivalents) are dissolved in 1-methyl-2-pyrrolidone (10 mL), and 2-chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (487.23 mg, 2.88 mmol, 1.5 equivalents) is added. The reaction mixture is stirred at 50 °C for 16 hours. 47% residual raw material is detected by LC-MS, and 36% of the target product is formed. 2-Chloro-1,3-dimethyl-4,5-dihydroimidazol-1-chloride (74.71 mg, 441.93 μmol, 0.23 equivalents) is added. The reaction mixture is stirred at 50 °C for 16 hours. 49% residual raw material is detected by LC-MS, and 38% of the target product is formed. Purification is carried out by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water (containing 0.05% formic acid) and acetonitrile (20%-40%) with decreasing polarity as the eluent) to obtain a pale yellow solid. It is detected by NMR that some formate salts are formed, and the crude product is purified by reverse-phase preparative liquid chromatography (Boston Prime C18 column, 5 μm silica, 30 mm diameter, 150 mm length, using a mixture of water (containing 0.05% ammonium hydroxide) and acetonitrile (32% - 52%) with decreasing polarity as the eluent) to obtain the white solid 4-amino-7-fluoro-N-(4-fluoro-2-methyl-pyrazol-3-yl)-1-methyl-N-[[6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl]methyl]imidazo[1,5-a]quinoxaline-8-carboxamide (Example 220) (155 mg, 276.26 μmol, 14.38% yield). LCMS: ES13685-1217-P1D1, (ESI) m / z = 556.1 [M+1] +, RT = 1.929 min NMR: ES13685 - 1217 - P1B, 1 1H NMR (400 MHz, DMSO - d6) δ 9.28 (br s, 1H), 8.10 (br s, 1H), 7.92 (br s, 1H), 7.82 (s, 1H), 7.73 (d, J = 9.76 Hz, 1H), 7.55 (s, 2H), 7.47 (br d, J = 8.63 Hz, 1H), 7.29 (br s, 1H), 7.11 (br d, J = 10.38 Hz, 1H), 5.19 (br s, 1H), 5.04 - 5.14 (m, 1H), 3.59 (s, 3H), 2.86 (br s, 3H). NMR: ES13685 - 1217 - P1B, 19F nmR (376 MHz, DMSO - d6) δ - 60.47 (s, 3F), - 118.15 (s, 1F), - 173.54 (s, 1F)

[0145] Synthesis of Example 287

Chemical Structure

[0146] Step 1: (E) - 4 - ((((dimethylamino)methylene)amino) - 7 - fluoro - 3 - methylimidazo[1,5 - a]quinoxaline - 8 - carbonyl chloride (Intermediate A7 - 1) To a stirred solution of 4-amino-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carboxylic acid (Intermediate A7) (270 mg, 1.04 mmol, 1 equiv) in DCM (3.00 mL) is added HCl / dioxane (4 M, 778 uL, 3 equiv). The mixture is stirred at 25 °C for 30 minutes. The reaction mixture is then concentrated and evaporated to dryness with toluene (10 mL × 3), the crude material is dissolved in DCM (3.00 mL) and cooled to 0 °C, and oxalyl chloride (790 mg, 6.23 mmol, 544 uL, 6 equiv is added dropwise at 0 °C) and DMF (75.8 mg, 1.04 mmol, 79.8 uL, 1 equiv) are added. The mixture is stirred at 25 °C for 12 hours. Complete consumption of the starting material is shown by LCMS and one major peak with the required mass is detected. The reaction mixture is concentrated, boiled with n-hexane (10 mL × 3) and dried under reduced pressure to give (E)-4-(((dimethylamino)methylene)amino)-7-fluoro-3-methylimidazo[1,5-a]quinoxaline-8-carbonyl chloride (Intermediate A7-1) (340 mg, crude product) as a yellow solid. LC-MS (ESI) m / z = 330.0 [M+H] +

[0147] Step 2: 4-Amino-7-fluoro-N-(1-methoxypropan-2-yl)-3-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 287) A solution of 1-methoxy-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)propan-2-amine (50.0 mg, 174 μmol, 1 eq) and DIEA (89.9 mg, 696 μmol, 121 μL, 4 eq) in THF (1.00 mL) is added with Int.24_COCl (63.9 mg, 191 μmol, 1.1 eq) at 0 °C. The mixture is stirred at 25 °C for 3 h. MeOH (1.00 mL) is added at 25 °C to quench the reaction mixture and concentrated under reduced pressure to give a residue. The residue is dissolved in MeOH (1 mL) and NH3 / MeOH (7M, 1 mL). The mixture is stirred at 70 °C for 2 h. LCMS shows complete consumption of the starting material and one major peak with the required mass is detected. The reaction mixture is concentrated under reduced pressure. The residue is purified by preparative HPLC (column: Waters Xbridge BEH C18 100×30 mm×10 μm, mobile phase: [water (NH4HCO3)-ACN], B%: 30% - 60%, 8 min, UV220 nm & 254 nm) to give 4-amino-7-fluoro-N-(1-methoxypropan-2-yl)-3-methyl-N-((6-(trifluoromethyl)imidazo[1,2-a]pyridin-2-yl)methyl)imidazo[1,5-a]quinoxaline-8-carboxamide (Example 287) as a yellow solid (58.1 mg, 109 μmol, 64.2% yield). 1 H NMR (400 MHz, DMSO-d6) δ 9.29 - 8.54 (m, 2H), 8.18 - 7.78 (m, 2H), 7.66 (br s, 1H), 7.38 (br d, J = 7.9 Hz, 1H), 7.13 (br s, 1H), 6.72 (br s, 2H), 4.95 - 4.39 (m, 2H), 4.22 - 3.82 (m, 1H), 3.52 (br s, 2H), 3.31 - 3.10 (m, 3H), 2.63 (s, 3H), 1.21 (br s, 3H) LC-MS (ESI) m / z = 530.2 [M + H] +

[0148] The following compounds are synthesized based on a general method, the structures of the products are shown in Table 1 and Table 2, and the characterization of the products is as follows.

[0149]

Table 2-1

[0150]

Table 2-2

[0151]

Table 2-3

[0152]

Table 2-4

[0153]

Table 2-5

[0154]

Table 2-6

[0155]

Table 2-7

[0156]

Table 2-8

[0157]

Table 2-9

[0158]

Table 2-10

[0159]

Table 2-11

[0160]

Table 2-12

[0161]

Table 2-13

[0162]

Table 2-14

[0163]

Table 2-15

[0164]

Table 2-16

[0165]

Table 2-17

[0166]

Table 2-18

[0167]

Table 2-19

[0168]

Table 2-20

[0169]

Table 2-21

[0170]

Table 2-22

[0171]

Table 2-23

[0172]

Table 2-24

[0173]

Table 2-25

[0174]

Table 2-26

[0175]

Table 2-27

[0176]

Table 2-28

[0177]

Table 2-29

[0178]

Table 2-30

[0179] Biological Test Example 2. In Vitro Inhibitory Proliferation Experiment of HCT116 and HCT116-MTAP-KO Cells

[0180] Experimental materials The HCT116 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences, and the MTAP gene was knocked out using CRISPR / Cas9 technology to obtain the HCT116-MTAP-KO cell line. McCoy's 5A medium (Gibco, catalog number 16600082), fetal bovine serum (Gibco, catalog number 10099141C), penicillin-streptomycin double antibody (Gibco, catalog number 15140122), trypsin (Gibco, catalog number 25200056), CellTiter-Glo detection kit (Promega, catalog number G7572), 384-well clear flat bottom black wall cell culture plate (Corning, catalog number 3764), ultra-micropipette device (Tecan, catalog number D300e), multifunctional microplate reader (Biotek, catalog number SynergyHTX)

[0181] Experimental methods 1. Cell culture: The culture conditions for HCT116 cells and HCT116-MTAP-KO cells are McCoy's 5A medium + 10% fetal bovine serum + 1% penicillin-streptomycin double antibody, always in the logarithmic growth phase, and it is confirmed that the cell viability exceeds 95%. 2. Preparation of compound concentration gradient: The test compound is added to a 384-well plate using an ultra-micropipette device, starting from 30 μM (HCT116 cells) or 3 μM (HCT116-MTAP-KO cells), diluted 3-fold with DMSO, for a total of nine concentrations, and three duplicate wells are set. 3. Compound-treated cells: The trypsin-digested HCT116 or HCT116-MTAP-KO cell suspension is added to the 384-well plate spotted with the test compound at 40 μL per well, that is, each well contains 100 cells, and the final DMSO concentration is 0.4%. The cell culture plate is placed in a 37°C, 5% carbon dioxide incubator and cultured for 6 days. 4. Detection: Add 20 μL of CellTiter-Glo reagent per well to the cell culture plate and incubate for 30 minutes while shaking at room temperature. Use a multifunctional microplate reader to detect the luminescence signal at 578 nm. 5. Data analysis: Use GraphPad Prism 8.0 software to fit the data using a four-parameter inhibitor-response model to obtain the IC 50 value (50% inhibitory concentration) of the test compound.

[0182] Measure the biological activities of several compounds by the experimental method. "A" represents IC 50 (nm) < 100, "B" represents 100 < IC 50 (nm) < 1000, "C" represents 1000 < IC 50 (nm) < 10000, as shown in Table 4, where the first column is the cell growth inhibition rate HCT116MTAP WT IC 50 (nm), and the second column is the cell growth inhibition rate HCT116-MTAP null IC 50 (nm).

[0183] The corresponding structures of the products to be tested are shown in Tables 1 and 2, and the results of the activity tests are as follows.

Table 3-1

[0184]

Table 3-2

[0185]

Table 3-3

[0186]

Table 3-4

[0187] All documents referred to in this invention are hereby incorporated by reference in this application as if each document was individually cited as a reference. Further, after reading the above teachings of this invention, those skilled in the art can make various changes or modifications to this invention, and these equivalent forms are also included within the scope defined by the appended claims of this application.

Claims

1. A compound as shown in the following formula I or II, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, wherein 【Chemical Formula 1】 [Chemical Formula 2] wherein Ra is 【Chemical Formula 3】 and W is O or S, X １ and X ２ are each independently selected from the group consisting of CR and N, and X ３ is N, L １ is selected from the group consisting of a chemical bond, -CHR-, -C(R)R- Ring A is selected from the group consisting of a substituted or unsubstituted 7- to 12-membered bridged ring (including a carbocyclic or heterocyclic ring), a substituted or unsubstituted 7- to 12-membered spiro ring (including a carbocyclic or heterocyclic ring), a substituted or unsubstituted 8- to 12-membered fused bicyclic heterocyclic group (including a carbocyclic or heterocyclic ring, preferably a 5-membered fused 6-membered ring), a substituted or unsubstituted 7- to 10-membered fused bicyclic heteroaryl group (preferably a 5-membered fused 6-membered ring), or Ring A is a substituted or unsubstituted 3- to 7-membered carbocyclic or heterocyclic ring, a substituted or unsubstituted 5- to 6-membered aromatic ring or heteroaromatic ring, Ring E is selected from the group consisting of a substituted or unsubstituted 3- to 7-membered monocyclic heterocyclic ring, a substituted or unsubstituted 7- to 12-membered bridged heterocyclic ring, a substituted or unsubstituted 7- to 12-membered spiro heterocyclic ring, a substituted or unsubstituted 8- to 12-membered fused polycyclic heterocyclic group (e.g., a fused bicyclic ring), R ８ is H, deuterium, a halogen, a cyano group, C ２ -C ６ -alkynyl group, -SF ５ , an amino group, a nitro group, a hydroxy group, a thiol group, an aldehyde group, a carboxy group, a substituted or unsubstituted or halogenated C １ -C ６ -alkyl group, a substituted or unsubstituted or halogenated C １ -C ６ -alkoxy group, or R ８ is 【Chemical Formula 4】 and R ８ ' is H, deuterium, halogen, cyano group, amino group, nitro group, hydroxy group, thiol group, aldehyde group, carboxy group, unsubstituted or halogenated C １ -C ６ alkyl group, substituted or unsubstituted benzene ring, substituted or unsubstituted 5- to 12-membered heteroaromatic ring, substituted or unsubstituted C ３ -C １０ carbocyclic ring (including the case of being saturated or partially unsaturated), substituted or unsubstituted 3- to 12-membered heterocyclic ring (including the case of being saturated or partially unsaturated), or R ８ ' is 【Chemical Formula 5】 and L ３ is selected from the group consisting of a chemical bond, -O-, -CHR-, -C(R)R-, a carbonyl group, S, and -NH- Ring B is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, a substituted or unsubstituted C ３ -C ６ carbocyclic ring (including the case of being saturated or partially unsaturated), a substituted or unsubstituted 3- to 7-membered heterocyclic ring (including the case of being saturated or partially unsaturated). R ２ and R ２ ’ are each independently selected from the group consisting of R ７ , -L ２ R ７ wherein L ２ is selected from the group consisting of -O-, -CHR-, -C(R)R-, wherein R ７ is hydrogen or none, substituted or unsubstituted C １ -C ６ alkyl group, substituted or unsubstituted C ６－１０ aromatic ring, substituted or unsubstituted 5- to 12-membered (preferably 5- to 6-membered or 8- to 10-membered) heteroaromatic ring, substituted or unsubstituted C ３ -C １０ carbocyclic ring (including saturated or partially unsaturated cases, including monocyclic, fused rings, spiro rings or bridged rings), substituted or unsubstituted 3- to 10-membered heterocyclic ring (including saturated or partially unsaturated cases, including monocyclic, fused rings, spiro rings or bridged rings), n is 0, 1, 2 or 3, R ３ is selected from the group consisting of H, deuterium, halogen, cyano group, substituted or unsubstituted C １ -C ６ alkyl group, R ４ and R ５ together with the ring atoms to which they are attached form a 5- to 12-membered saturated or unsaturated ring, and said ring may or may not be substituted, R is H, deuterium, a halogen, a substituted or unsubstituted C １ -C ４ alkyl group, a substituted or unsubstituted C １ -C ４ alkoxy group, a substituted or unsubstituted C ３ -C ６ cycloalkyl group, and Unless otherwise specified, in the above formulas, the substitution means that the hydrogen atom on the corresponding group is deuterium, tritium, halogen, hydroxy group, carboxy group, thiol group, benzyl group, C １ -C １２ alkoxycarbonyl group, C １ -C ６ aldehyde group, amino group, C １ -C ６ amide group, nitro group, cyano group, unsubstituted or halogenated C １ -C ６ alkyl group, unsubstituted or halogenated C １ -C ６ alkyl-O-C １ -C ６ alkyl-, unsubstituted or halogenated C １ -C ６ alkyl-O-C １ -C ６ alkyl-O-, unsubstituted or halogenated C １ -C ６ alkylene-OH, unsubstituted or halogenated C ３ -C ８ cycloalkyl group, C ２ -C １０ alkenyl group, unsubstituted or halogenated C １ -C ６ alkoxy group, C １ -C ６ alkyl-amine group, C ６ -C １０ aryl group, five- or six-membered heteroaryl group, five- or six-membered non-aromatic heterocyclic group, -O-(C ６ -C １０ aryl), -O-(five- or six-membered heteroaryl), C １ -C １２ alkylaminocarbonyl group, unsubstituted or halogenated C ２ -C １０ acyl group, sulfonyl (-SO ２ -OH), phosphoryl (-PO ３ -OH), unsubstituted or halogenated C １ -C ４ alkyl-S(O) ２ -, unsubstituted or halogenated C １ -C ４ alkyl-SO—, —SF ５ A compound as shown in formula I or II, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof, characterized by being substituted by one or more substituents selected from the group consisting of

2. Ra is selected from the group consisting of 【Chemical Formula 6】 Here, R ９ is selected from the group consisting of deuterium, tritium, halogen, hydroxy group, carboxy group, unsubstituted or halogenated C １ -C ６ alkyl group, unsubstituted or halogenated C １ -C ６ alkoxy group, unsubstituted or substituted C １ -C ６ alkyl-OH, -NH(unsubstituted or halogenated C １ -C ６ alkyl group), -N(unsubstituted or halogenated C １ -C ６ alkyl group), ２ m is selected from 0, 1, 2 or 3, and preferably, R ９ is selected from the group consisting of deuterium, tritium, halogen, unsubstituted or halogenated C １ -C ６ alkyl group, characterized in that The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

3. L １ is -CHR-, -C(R)R-, and ring A is selected from the group consisting of a substituted or unsubstituted 8- to 12-membered fused bicyclic heterocyclic group and a substituted or unsubstituted 7- to 10-membered fused bicyclic heteroaryl group, R ８ is H, halogen, cyano group, amino group, C ２ -C ６ alkynyl group, SF ５ -, hydroxy group, thiol group, aldehyde group, carboxy group, unsubstituted or halogenated C １ -C ６ alkyl group, 【Chemical Formula 7】 selected from the group consisting of, and the B ring is a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, a substituted or unsubstituted C ３ -C ６ carbocyclic ring, selected from the group consisting of a substituted or unsubstituted 3- to 6-membered heterocyclic ring, L ３ is selected from the group consisting of a chemical bond, -O-, -CHR-, a carbonyl group, S, or -NH- The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

4. R ２ is selected from the group consisting of R ７ , -L ２ R ７ wherein L ２ is selected from the group consisting of -O-, -CHR-, a carbonyl group, S, -NH-, wherein R ７ is a substituted or unsubstituted C １ -C ６ alkyl group, a substituted or unsubstituted C ６－１０ aromatic ring, a substituted or unsubstituted 5- to 12-membered heteroaromatic ring, characterized in that it is selected from the group consisting of The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

5. R ２ is an ortho-substituted 5- or 6-membered heteroaromatic ring as shown by the following formula, 【Chemical 8】 Here, R １０ is a substituent located at the binding site, which is selected from the group consisting of hydrogen, deuterium, halogen, halogenated or non-halogenated C １ -C ３ alkyl group, halogenated or non-halogenated C １ -C ３ alkoxy group Ring D is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, preferably Ring D is selected from the group consisting of the following, characterized in that 【Chemical Formula 9】 The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

6. L １ is -CH ２ -, -CH(CH ３ )-, and ring A is selected from the group consisting of the following: 【Chemical 10】 Here, ring C is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, a substituted or unsubstituted C ３ -C ６ carbocyclic ring (including the case of being saturated or partially unsaturated), a substituted or unsubstituted 3- to 6-membered heterocyclic ring (including the case of being saturated or partially unsaturated). Or Ring A is selected from the group consisting of the following, 【Chemical Formula 11】 R ８ is a halogenated or non-halogenated C １ -C ６ alkyl group, or 【Chemical Formula 12】 and ring B is selected from the group consisting of a substituted or unsubstituted benzene ring, a substituted or unsubstituted 5- to 6-membered heteroaromatic ring, a substituted or unsubstituted C ３ -C ６ carbocyclic ring, and a substituted or unsubstituted 3- to 6-membered heterocyclic ring, and L ３ is selected from the group consisting of a chemical bond, -O-, -CHR-, a carbonyl group, S, or -NH- The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

7. R ２ is selected from the group consisting of R ７ , -(CHR)R ７ , where R ７ is hydrogen or none, substituted or unsubstituted C １ -C ６ alkyl group, substituted or unsubstituted C ６－１０ aromatic ring, substituted or unsubstituted 5- to 12-membered heteroaromatic ring, substituted or unsubstituted C ３ -C ８ carbocyclic ring (including the case of saturated or partially unsaturated, including monocyclic, fused ring, spiro ring or bridged ring), substituted or unsubstituted 3- to 8-membered heterocyclic ring (including the case of saturated or partially unsaturated, including monocyclic, fused ring, spiro ring or bridged ring), characterized in that it is selected from the group consisting of The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

8. The compound has a structure as shown in the following formula, 【Chemical 13】 Here, Q is O, NH, CH ２ , or a chemical bond (i.e., 【Chemical 14】 is a five-membered ring), and R ８ is defined as described above, R ８ａ and R ８ｂ are each independently selected from H, or R ８ａ and R ８ｂ together with the carbon atom to which they are attached form a 4- to 7-membered carbocyclic or heterocyclic ring, and R ８ａ and R ８ｂ When each of them is independently H, R ８ａ or R ８ｂ is R ８ can be arbitrarily substituted by R ８ａ and R ８ｂ When they combine with the carbon atom to which they are attached to form a 4- to 7-membered carbocyclic or heterocyclic ring, R ８ is characterized in that it can be located on the carbocyclic or heterocyclic ring The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

9. R ３ is selected from the group consisting of H, deuterium, halogen, cyano group, substituted or unsubstituted C １ -C ６ alkyl group, characterized by The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

10. R ２ is a substituted or unsubstituted 5- to 7-membered heteroaromatic ring, and ring A is selected from the group consisting of a substituted or unsubstituted 5- to 6-membered aromatic or heteroaromatic ring, a substituted or unsubstituted 7- to 10-membered fused bicyclic heteroaryl group, R ８ is CF ３ characterized in that The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

11. The compound is characterized in that it is selected from the group consisting of 【Chemical Formula 15-1】 【Chemical Formula 15-2】 【Chemical Formula 15-3】 【Chemical 15-4】 【Chemical Formula 15-5】 【Chemical Formula 15-6】 【Chemical 15-7】 【Chemical Formula 15-8】 【Chemical Formula 15-9】 【Chemical 15-10】 [[Chemical Formula 15-11]] 【Chemical Formula 15-12】 【Chemical 15-13】 【Chemical Formula 15-14】 【Chemical 15-15】 【Chemical Formula 15-16】 【Chemical Formula 15-17】 【Chemical Formula 15-18】 [Chemical Formula 15-19] 【Chemical 15-20】 【Chemical 15-21】 【Chemical Formula 15-22】 【Chemical 15-23】 The compound according to claim 1, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

12. Selected from the group consisting of 【Chemical Formula 16-1】 【Chemical 16-2】 A compound, or a pharmaceutically acceptable stereoisomer, salt or deuterated product thereof.

13. A pharmaceutical composition, The pharmaceutical composition comprises a therapeutically effective amount of one or more compounds according to any one of claims 1 to 12, a pharmaceutically acceptable salt, racemate, optical isomer, stereoisomer or tautomer thereof, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary substances and / or diluents. The pharmaceutical composition is characterized by the above.

14. Use of a compound according to any one of claims 1 to 12, a racemate, stereoisomer or pharmaceutically acceptable salt thereof, in the preparation of a drug for treating or preventing a disease associated with an abnormality or abnormal expression at the gene level of PRMT5 (for example, a mutation, deletion of the corresponding nucleic acid, or an abnormality at the MTAP gene level, or ectopic production, fusion or overexpression of methyltransferase).

15. The disease is characterized in that it is selected from the group consisting of a disease or pathological ovarian cancer, esophageal cancer, lung cancer, lymphoma, glioblastoma, colorectal cancer, melanoma, gastric cancer, pancreatic cancer or bladder cancer The use according to claim 14.

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