A pyrimidopyridone derivative as an SOS1 inhibitor, its preparation method and uses

Abstract

A pyrimidopyridone derivative as an SOS1 inhibitor, its preparation method, and its uses. The structure of the pyrimidopyridone derivative is shown in formula (I), which has a significant inhibitory effect on the RAS signaling pathway and is used for the treatment and / or prevention of cancer, including pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer, and cholangiocarcinoma.

Description

Technical Field

[0001] This invention belongs to the technical field of pyrimidopyridone derivatives, specifically relating to a pyrimidopyridone derivative as an SOS1 inhibitor, its preparation method, and its uses. Background Technology

[0002] The RAS family of proteins belongs to a class of small GTPases, comprising three subfamilies: KRAS, NRAS, and HRAS. Mutated RAS genes are important oncogenes, found in 20-30% of human tumors, particularly pancreatic, colorectal, and lung cancers. All RAS protein subtypes exist in a balance between a GTP-bound activated state and a GDP-bound inactive state. GTPase-activating proteins (GAPs) promote the conversion of GTP to GDP, thus inactivating RAS proteins. Conversely, guanine nucleotide exchange factors (GEFs) promote the release of GDP and the binding of GTP, thus activating RAS proteins. Activation of RAS proteins promotes cell proliferation, apoptosis evasion, and metabolic remodeling through the RAS-RAF-MEK-ERK and RAS-PI3K-PDK1-AKT signaling pathways, thereby contributing to tumorigenesis and development.

[0003] SOS1 (son of sevenless 1) is a key guanine nucleotide exchange factor (GEF) that binds to RAS proteins, promoting the binding of RAS proteins to GTP and activating the RAS protein. Recent studies have found that SOS1 inhibitors alone can inhibit the growth of various RAS mutant cells and can also produce a synergistic effect with MEK inhibitors, significantly inhibiting KRAS-driven tumors. 1-2 The development of SOS1 inhibitors has become a research hotspot, with many patents reporting SOS1 inhibitors of different structural types, such as WO2018172250, WO2019201848, WO2018115380, WO2019122129, WO2020173935, WO2020180768, and WO2020180770.

[0004] However, there are still uncertainties regarding the efficacy, safety, or selectivity of the compounds and experimental drugs disclosed in these existing technologies. Therefore, it is necessary to study and develop new selective SOS1 inhibitors.

[0005] References:

[0006] 1. Hillig et al. Discovery of poetent SOS1 inhibitors that block RASactivation via disruption of the RAS-SOS1 interaction. PNAS. 116, 2251-2560 (2019).

[0007] 2. Hofmann et al. BI-3406, a potent and selective SOS1:: KRAS interactioninhibitor, is effective in KRAS-driven cancers through combined MEKinhibition. Cancer Discov. CD-20-0142 (2020). Summary of the Invention

[0008] In order to solve the above-mentioned problems of the prior art, the present invention aims to provide a pyrimidine pyridinone derivative, its pharmaceutically acceptable salt, its tautomer or its stereoisomer, to screen for compounds that have excellent performance in terms of efficacy, safety and selectivity as SOS1 inhibitors.

[0009] Another object of the present invention is to provide a method for preparing the said derivative, its pharmaceutically acceptable salt, its tautomer or its stereoisomer.

[0010] To achieve this objective, the present invention employs the following technical solution:

[0011] In a first aspect, the present invention provides a pyrimidopyridone derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof, or a stereoisomer thereof, the structure of which is shown in formula (I):

[0012]

[0013] Where: R 1 Selected from hydrogen or C1-C3 alkyl; preferably hydrogen or methyl;

[0014] R 2 Selected from hydrogen or C1-C3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclic groups, wherein the C1-C3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclic groups are optionally surrounded by 1-3 R groups. 21 Replaced;

[0015] R 21Selected from C1-C3 alkyl, hydroxyl, halogen, cyano, amino, C1-C3 alkoxy, or =O;

[0016] L may be absent or selected from O, NH or N-(C1-C3 alkyl);

[0017] R 3 Selected from H, C1-C3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclic groups, wherein the C1-C3 alkyl, 3-7 membered cycloalkyl, and 4-7 membered heterocyclic groups are optionally surrounded by 1-3 R groups. 31 Replaced;

[0018] R 31 Selected from C1-C3 alkyl, C1-C3 haloalkyl, hydroxyl, halogen, cyano, -NR a R b C1-C3 alkoxy groups, =O, -NHCOR 32 or -COR 32 ;

[0019] R a Selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or 3-6 membered cycloalkyl;

[0020] R b Selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or 3-6 membered cycloalkyl;

[0021] R 32 Selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclic groups;

[0022] AR is selected from 6-10 aryl or 5-10 heteroaryl, wherein the aryl or heteroaryl is optionally surrounded by 1-4 R 4 Replaced;

[0023] R 4 Selected from H, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy-C1-C3 alkyl, hydroxy-C1-C3 haloalkyl, 3-6 membered cycloalkyl, 4-7 membered heterocyclic group, -OR a -NR a R b 6-10 yuan aryl or 5-10 yuan heteroaryl, wherein the 6-10 yuan aryl or 5-10 yuan heteroaryl is optionally surrounded by 1-4 R. c Replaced;

[0024] R c Selected from H, halogen, C1-C3 alkyl, C1-C3 haloalkyl, hydroxy-C1-C3 alkyl, hydroxy-C1-C3 haloalkyl, 3-6 membered cycloalkyl, 4-7 membered heterocyclic group, -OR a -NRa R b NR a R b -C1-C4 alkyl, NR a R b -C1-C4 haloalkyl; the heterocyclic or heteroaryl group in formula (I) has 1-3 heteroatoms selected from one or more of oxygen, nitrogen and sulfur.

[0025] Preferably, the structure of the pyrimidinepyridone derivative is shown in formula (II):

[0026]

[0027] Among them, R 1 R 2 R 3 and R 4 It has the same limited range as above; n = 1-4 (e.g., n = 1, n = 2, n = 3, n = 4);

[0028] Preferably, in the compound of formula (II), the phenyl group is optionally surrounded by 1-4 R groups. 4 Replaced when the R 4 When the number of R is 2-4, the R 4 They can be the same or different;

[0029] And / or, when the R 4 When R is a C1-C3 haloalkyl group, 4 The number is 1-3, when there are two or more R. 4 At that time, the R mentioned 4 They can be the same or different;

[0030] And / or, when the R 4 When it is a C1-C3 haloalkyl group, the halogen atom is fluorine;

[0031] And / or, when the R 4 When it is halogen, the R 4 The number is 1-3, when there are two or more R. 4 At that time, the R mentioned 4 They can be the same or different;

[0032] And / or, when the R 4 When it is a halogen, the halogen atom is fluorine;

[0033] And / or, when the R 4 For -NR a R b At that time, the R mentioned 4 The number is 1-3, when there are two or more R. 4 At that time, the R mentioned4 They can be the same or different;

[0034] And / or, when the R 4 For -NR a R b At that time, the R mentioned a and R b They can be the same or different;

[0035] and / or R 3 It is H, C1-C3 alkyl, 3-7 membered cycloalkyl, or 4-7 membered heterocyclic, wherein the 4-7 membered heterocyclic group is optionally surrounded by 1-3 R groups. 31 Replaced;

[0036] And / or, when R 3 To be arbitrarily assigned to 1-3 R 31 The 4-7 membered heterocyclic group replaced, the R 31 When there are 2-3, R 31 Same or different;

[0037] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the substituted 4-7 membered heterocyclic group is used, the heterocyclic group contains 1-2 heteroatoms;

[0038] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the substituted 4-7 membered heterocyclic group is used, the heteroatom of the heterocyclic group is nitrogen and / or oxygen;

[0039] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the substituted 4-7 membered heterocyclic group has two heteroatoms, the two heteroatoms may be the same or different.

[0040] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the 4-7 membered heterocyclic group is replaced, the R 31 Selected from C1-C3 alkyl, C1-C3 haloalkyl, hydroxyl, halogen, cyano, -NR a R b C1-C3 alkoxy groups, =O, -NHCOR 32 or -COR 32 ;

[0041] R a Selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or 3-6 membered cycloalkyl;

[0042] R bSelected from H, C1-C3 alkyl, C1-C3 haloalkyl, or 3-6 membered cycloalkyl;

[0043] R 32 It is selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclic groups.

[0044] Preferably, the structure of the pyrimidinepyridone derivative is shown in formula (III):

[0045]

[0046] Among them, R 1 R 2 R 3 and R 4 It has the same limited range as above; n = 1-4 (e.g., n = 1, n = 2, n = 3, n = 4).

[0047] More preferably, for compounds of formula (III), the phenyl group is optionally surrounded by 1-4 R groups. 4 Replaced when the R 4 When the number of R is 2-4, the R 4 They can be the same or different;

[0048] And / or, when the R 4 When R is a C1-C3 haloalkyl group, 4 The number is 1-3, when there are two or more R. 4 At that time, the R mentioned 4 They can be the same or different;

[0049] And / or, when the R 4 When it is a C1-C3 haloalkyl group, the halogen atom is fluorine;

[0050] And / or, when the R 4 When it is halogen, the R 4 There are 1-2 R's. 4 At that time, the R mentioned 4 They can be the same or different;

[0051] And / or, when the R 4 When it is a halogen, the halogen atom is fluorine;

[0052] And / or, when the R 4 For -NR a R b At that time, the R mentioned 4 The number is 1-3, when there are two or more R. 4 At that time, the R mentioned 4 They can be the same or different;

[0053] And / or, when the R 4 For -NR a R b At that time, the R mentioned a and R b They can be the same or different;

[0054] and / or R 3 It is H, C1-C3 alkyl, 3-7 membered cycloalkyl, or 4-7 membered heterocyclic, wherein the 4-7 membered heterocyclic group is optionally surrounded by 1-3 R groups. 31 Replaced;

[0055] And / or, when R 3 To be arbitrarily assigned to 1-3 R 31 The 4-7 membered heterocyclic group replaced, the R 31 When there are 2-3, R 31 Same or different;

[0056] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the substituted 4-7 membered heterocyclic group is used, the heterocyclic group contains 1-2 heteroatoms;

[0057] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the substituted 4-7 membered heterocyclic group is used, the heteroatom of the heterocyclic group is nitrogen and / or oxygen;

[0058] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the substituted 4-7 membered heterocyclic group has two heteroatoms, the two heteroatoms may be the same or different.

[0059] And / or, the R 3 To be arbitrarily assigned to 1-3 R 31 When the 4-7 membered heterocyclic group is replaced, the R 31 Selected from C1-C3 alkyl, C1-C3 haloalkyl, hydroxyl, halogen, cyano, -NR a R b C1-C3 alkoxy groups, =O, -NHCOR 32 or -COR 32 ;

[0060] R a Selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or 3-6 membered cycloalkyl;

[0061] R b Selected from H, C1-C3 alkyl, C1-C3 haloalkyl, or 3-6 membered cycloalkyl;

[0062] R 32 It is selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclic groups.

[0063] Preferably, the structure of the pyrimidinepyridone derivative is shown in formula (IV):

[0064]

[0065] Among them, R 1 R 2 R3, R c And L have the same limited range as above; m = 1-4 (e.g., m = 1, m = 2, m = 3, m = 4).

[0066] More preferably, in the compound of formula (IV), L is selected from O, NH or NCH3.

[0067] More preferably, the pyrimidinepyridone derivative is selected from any one of the following structures:

[0068]

[0069]

[0070]

[0071]

[0072]

[0073] In a second aspect, the present invention provides a method for preparing the pyrimidopyridone derivatives as described in the first aspect, pharmaceutically acceptable salts thereof, tautomers thereof, or stereoisomers thereof, selected from one of the following two schemes:

[0074] Option 1

[0075] The method for preparing the compound of general formula (I) of the present invention, or its stereoisomers, tautomers, or pharmaceutical salts thereof, includes the following steps:

[0076] Preparation of the key intermediate (IA):

[0077]

[0078] In the first step, aromatic compounds of general formula (I-1) are cross-coupled by metal catalysis to obtain compounds of general formula (I-2);

[0079] In the second step, the compound of general formula (I-2) reacts under catalytic conditions to give the chiral sulfonamide compound of general formula (I-3);

[0080] The third step involves reducing the compound of general formula (I-3) with a metal reducing agent to obtain the chiral compound of general formula (I-4);

[0081] In the fourth step, the compound of general formula (I-4) is cleaved under acidic conditions by sulfonamide to give the chiral benzylamine compound of general formula (IA).

[0082] Preparation of the key intermediate (IB), Method 1:

[0083]

[0084] In the first step, the compound of general formula (I-5) and tert-butyl bromoacetate were subjected to a substitution reaction under basic conditions to obtain the compound of general formula (I-6);

[0085] The second step involves detert-butylating the compound of general formula (I-6) under acidic conditions to obtain the compound of general formula (I-7);

[0086] The third step involves esterifying the compound of general formula (I-7) with an alcohol under acidic conditions to obtain the compound of general formula (I-8).

[0087] In the fourth step, the compound of general formula (I-9) and the amine of general formula (I-10) undergo a substitution reaction to give the compound of general formula (I-11);

[0088] Fifth step: The compound of general formula (I-11) is halogenated to give the compound of general formula (I-12);

[0089] Step 6: The compound of general formula (I-12) and boric acid or boron ester compound are reacted under alkaline conditions in the presence of a metal catalyst and ligands via the Suzuki reaction to obtain the compound of general formula (I-13).

[0090] Step 7: Oxidation of the double bond in the compound of general formula (I-13) yields the compound of general formula (I-14);

[0091] Step 8: The compound of general formula (I-14) and the compound of general formula (I-8) are subjected to alkaline conditions to give the compound of general formula (IB).

[0092] Preparation of the key intermediate (IB), Method 2:

[0093]

[0094] In the first step, a compound of general formula (I-15) and an amine of general formula (I-10) are subjected to a substitution reaction to give a compound of general formula (I-16);

[0095] In the second step, the compounds of general formula (I-16) and general formula (I-8) are subjected to alkaline conditions to obtain the compound of general formula (IB).

[0096] Preparation of general formula (I):

[0097]

[0098] In the first step, compounds of general formula (IB) and general formula (IA) undergo a substitution reaction to obtain compounds of general formula (I).

[0099] Among them, X, X 1 X 2 and X 3 X is a halogen; X is preferably bromine; X 1 X 2 Chlorine is preferred; X 3 Iodine is preferred; R 1 R 2 R 3 AR and L have the same limited range as described above.

[0100] Option 2

[0101] The method for preparing the compound of general formula (I) of the present invention, or its stereoisomers, tautomers, or pharmaceutical salts thereof, includes the following steps:

[0102]

[0103] In the first step, the compound of general formula (I-12) and methyl acrylate are reacted via the Heck reaction under alkaline conditions and with a metal catalyst to obtain the compound of general formula (I-17);

[0104] The second step involves the intramolecular formation of amides from the compound of general formula (I-17) under alkaline conditions to yield the compound of general formula (I-18).

[0105] The third step involves halogenating the compound of general formula (I-18) to obtain the compound of general formula (I-19).

[0106] Fourth step: The compound of general formula (I-19) is demethylated under acidic conditions to give the compound of general formula (I-20);

[0107] Fifth step: The compound of general formula (I-20) reacts with 2,4,6-triisopropylbenzenesulfonyl chloride or phosphorus oxychloride under alkaline conditions to give the compound of general formula (I-21);

[0108] Step 6: The compound of general formula (I-21) and the compound of general formula (IA) react under basic conditions to give the compound of general formula (I-22);

[0109] In the seventh step, the compounds of general formula (I-22) and general formula (I-23) are reacted under alkaline conditions in the presence of a metal catalyst and ligands via the Suzuki reaction to give the compound of general formula (I).

[0110] Among them, X 1 X 3 and X 4 Halogen; X 1 Chlorine is preferred; X 3 Iodine is preferred; X 4 Preferably bromine; X 5 Halogen or W is R 1 R 2 R 3 AR and L have the same limited range as described above.

[0111] In the above preparation method,

[0112] The reagents providing alkaline conditions are selected from organic or inorganic bases. The organic bases are one or more of triethylamine, N,N-diisopropylethylamine, n-butyllithium, diisopropylaminolithium, bis(trimethylsilylaminolithium), sodium tert-butoxide, sodium methoxide, and potassium tert-butoxide. The inorganic bases are one or more of sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium bicarbonate, and lithium hydroxide.

[0113] The reagents that provide acidic conditions are one or more of the following: hydrogen chloride, a 1,4-dioxane solution of hydrogen chloride, a methanol solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, and phosphoric acid.

[0114] The metal catalyst is one or more of palladium / carbon, Raney nickel, tetra-triphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride (Pd(dppf)Cl2), [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex, bis(triphenylphosphine)palladium dichloride (Pd(PPh3)Cl2), and tris(dibenzylideneacetone)palladium (Pd2(dba)3);

[0115] The ligand is one or more of 2-bicyclohexylphosphine-2,6′-dimethoxybiphenyl (SPhos), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (XantPhos), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (XPhos), 2-dicyclohexylphosphine-2′-(N,N-dimethylamine)-biphenyl (DavePhos), 1,1′-bis(diphenylphosphine)ferrocene (Dppf), and 1,1′-binaphine-2,2′-bisdiphenylphosphine (BINAP), preferably 1,1′-binaphine-2,2′-bisdiphenylphosphine (BINAP);

[0116] The reducing agent is one or more of sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, and lithium aluminum hydride.

[0117] The oxidizing agent is one or more of potassium permanganate, manganese dioxide, potassium dichromate, sodium dichromate, and potassium osmium tetroxide.

[0118] The above reaction is preferably carried out in a solvent, which is one or more selected from N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, 1,4-dioxane, water, tetrahydrofuran, dichloromethane, 1,2-dichloroethane, methanol, ethanol, toluene, petroleum ether, ethyl acetate, n-hexane, and acetone.

[0119] Thirdly, the present invention provides a pharmaceutical composition comprising, as described in the first aspect, pyrimidine-pyridone derivatives and / or stereoisomers, tautomers, pharmaceutically acceptable salts; and pharmaceutically acceptable carriers and / or excipients and / or sustained-release agents.

[0120] In this invention, the term "containing" indicates that various ingredients may be used together in the mixtures or compositions of this invention. Therefore, the terms "consistent with..." and "composed of..." are included in the term "containing".

[0121] In this invention, a "pharmaceuticalally acceptable" ingredient is a substance that is suitable for humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., has a reasonable benefit / risk ratio.

[0122] In this invention, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent, or excipient used to deliver the active substance of this invention or its physiologically acceptable salt to animals or humans. The carrier can be liquid or solid.

[0123] In this invention, the pharmaceutical composition contains a safe and effective amount (e.g., 0.001-99.9 parts by weight, more preferably, 0.01-99 parts by weight, even more preferably 0.1-90 parts by weight) of a compound of formula (I) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 parts by weight.

[0124] Alternatively, the pharmaceutical composition of the present invention contains 0.001-99.9 wt%, more preferably 0.01-99 wt%, more preferably 0.1-90 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 wt%.

[0125] In another preferred embodiment, the preferred ratio of the compound of formula (I) to a pharmaceutically acceptable carrier, excipient or sustained-release agent is that the active ingredient of formula (I) accounts for more than 65% of the total weight, and the remainder accounts for 0.5-40% of the total weight, or more preferably 1-20%, or most preferably 1-10%.

[0126] Various formulations of the pharmaceutical compositions of the present invention contain, per unit dose, 0.05 mg to 500 mg, preferably 0.5 mg to 200 mg, more preferably 0.1 mg to 100 mg of the compound of formula (I), an enantiomer, a racemic mixture, a pharmaceutically acceptable salt, or a mixture thereof.

[0127] When the pharmaceutical composition contains additional active pharmaceutical ingredient for treating or preventing cancer, the amount of such active ingredient is typically the conventional amount or lower in the prior art.

[0128] The pharmaceutical compositions of the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions, and aerosols, wherein the compound of formula (I) can be present in a suitable solid or liquid carrier or diluent. The pharmaceutical compositions of the present invention can also be stored in suitable sterile instruments for injection or infusion. The pharmaceutical compositions may also contain odorants, flavorings, etc.

[0129] The compounds of formula (I) or pharmaceutical compositions comprising formula (I) of the present invention may be administered clinically to mammals (including humans) via routes of administration such as oral, nasal, skin, lung, or gastrointestinal tract. Oral administration is preferred. The preferred daily dose is 0.5 mg to 200 mg / kg body weight, taken once or in divided doses. Regardless of the method of administration, the optimal dose for an individual should be determined based on the specific treatment. Generally, a low dose is started, and the dose is gradually increased until the most suitable dose is found.

[0130] The effective dose of the active ingredient used can vary depending on the compound used, the mode of administration, and the severity of the disease being treated. However, generally, satisfactory effects are obtained when the compounds of the present invention are administered daily at a dose of about 1-300 mg / kg of animal body weight, preferably in 1-3 separate doses daily, or in a sustained-release form. For most large mammals, the total daily dose is about 5-1000 mg, preferably about 10-500 mg. Suitable oral dosage forms comprise about 1-200 mg of the active compound, closely mixed with a pharmaceutically acceptable solid or liquid carrier. This dosage regimen can be adjusted to provide optimal therapeutic response. For example, due to the urgency of the treatment condition, several separate doses may be administered daily, or the dose may be reduced proportionally.

[0131] The compound or its pharmaceutically acceptable salts and combinations thereof may be administered orally, as well as intravenously, intramuscularly, or subcutaneously. From the standpoint of ease of preparation and administration, the preferred pharmaceutical compositions are solid compositions, particularly tablets and solid-filled or liquid-filled capsules. Oral administration of the pharmaceutical composition is preferred.

[0132] Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose, and kaolin, while liquid carriers include sterile water, polyethylene glycol, nonionic surfactants, and edible oils (such as corn oil, peanut oil, and sesame oil), provided they are suitable for the characteristics of the active ingredient and the desired specific route of administration. Adjuvants commonly used in the preparation of pharmaceutical compositions may also be advantageously included, such as flavoring agents, colorings, preservatives, and antioxidants such as vitamin E, vitamin C, BHT, and BHA.

[0133] The active compounds, or their pharmaceutically acceptable salts and combinations thereof, may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds (as free bases or pharmaceutically acceptable salts) can also be prepared in water with a suitable surfactant (such as hydroxypropyl cellulose). Dispersions can also be prepared in glycerol, liquids, polyethylene glycol, and mixtures thereof in oils. Under normal storage and use conditions, these formulations contain preservatives to prevent microbial growth.

[0134] Suitable injectable drug forms include: sterile aqueous solutions or dispersions and sterile powders (for the ad hoc preparation of sterile injectable solutions or dispersions). In all cases, these forms must be sterile and must be fluid to facilitate dispensing via syringe. They must be stable under manufacturing and storage conditions and must be protected against contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing substances such as water, alcohols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0135] The compound represented by formula (I), or its pharmaceutically acceptable salt, and combinations thereof, may also be administered in combination with other active ingredients or drugs for the treatment or prevention of chronic pain disorders. When two or more drugs are administered in combination, they generally have a better effect than when either drug is administered alone.

[0136] Fourthly, the present invention provides the use of pyrimidopyridone derivatives, stereoisomers, tautomers, pharmaceutically acceptable salts, or pharmaceutical compositions as described in the first aspect, alone or in combination with a KRAS inhibitor, in the preparation of a medicament for treating cancer or in the preparation of an SOS1 inhibitor; preferably, the cancers include, but are not limited to, astrocytic carcinoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, pancreatic cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer, thyroid cancer, sarcoma, renal cancer, and cholangiocarcinoma; more preferably, the cancers include, but are not limited to, pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer, and cholangiocarcinoma; and / or the KRAS inhibitor is a KRAS G12C, KRAS G12V, KRAS G12S, or KRAS G12D inhibitor; and / or a MEK signaling pathway inhibitor such as RAF, MEK, or ERK1 / 2 inhibitor.

[0137] Preferably, the dosage range of the pyrimidine-pyridone derivatives, their stereoisomers, tautomers, pharmaceutically acceptable salts, or pharmaceutical compositions as described in the first aspect is selected from 10-100 mg / kg, preferably 25-50 mg / kg; the dosage range of the KRAS G12C inhibitor is selected from 3-100 mg / kg, preferably 5-30 mg / kg; and the dosage range of the MEK inhibitor is selected from 0.1-0.2 mg / kg, preferably 0.125 mg / kg.

[0138] Preferably, the KRAS G12C inhibitor is MRTX849, and the MEK inhibitor is Trametinib.

[0139] The use as described in the fourth aspect, wherein the cancer is a cancer associated with the RAS family; preferably, cancers associated with KRAS, HRAS, or NRAS G12C, KRAS, HRAS, or NRAS G12D, KRAS, HRAS, or NRAS G12V, KRAS, HRAS, or NRAS G12S, KRAS, HRAS, or NRAS G12A, KRAS, HRAS, or NRAS G13D, KRAS, HRAS, or NRAS G13C, KRAS, HRAS, or NRAS Q61L, KRAS, HRAS, or NRAS A146T, KRAS, HRAS, or NRAS A146V, or KRAS, HRAS, or NRAS A146P.

[0140] On the other hand, methods for treating cancer are provided, including providing a cancer patient with a therapeutically effective amount of formula (I), its pharmaceutically acceptable salt, or a pharmaceutical composition containing the compound or its pharmaceutically acceptable salt. In one embodiment, the cancer is a RAS family-related cancer. In one embodiment, the cancer is an SOS-L-related cancer. In one embodiment, the cancer is an NF-1 / NF-2-related cancer.

[0141] The compositions and methods provided by this invention can be used to treat a variety of cancers, including prostate cancer, breast cancer, brain cancer, skin cancer, cervical cancer, and testicular cancer. More specifically, the cancers that can be treated by the compositions and methods of this invention include, but are not limited to, tumor types such as astrocytic carcinoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, stomach cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer, thyroid cancer, and sarcoma.More specifically, these compounds can be used to treat: Heart: sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma; Lung: bronchial carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroma, mesothelioma; Gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagon, gastrinoma, carcinoid, VIPoma), small intestine (adenocarcinoma, lymphoma, carcinoid, Kaposi's sarcoma, leiomyosarcoma, hemangioma, lipoma, neurofibroma, fibroma). Colorectal cancer (adenocarcinoma, tubular adenoma, villonoma, hamartoma, leiomyoma); Genitourinary tract: Kidney (adenocarcinoma, nephroblastoma, lymphoma, leukemia), Bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), Prostate (adenocarcinoma, sarcoma), Testis (seminomatous seminoma, teratoma, embryonal carcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); Liver: Hepatocellular carcinoma, bile duct carcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bile duct: Gallbladder carcinoma, ampullary carcinoma, bile duct carcinoma; Bone: Osteosarcoma, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell carcinoma) Tumors: Multiple myeloma, malignant giant cell tumor, chordoma, chronic bone tumor (osteochondrosarcoma), benign tumors, chondroblastoma, chondromycinoma, osteoid osteoma, and giant cell tumor; Nervous system: Skull (osteoma, hemangioma, granuloma, xanthoma, osteitis), Meninges (meningioma, meningeal sarcoma, glioma), Brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal tumor), glioblastoma pleomorphic, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal fibroma, meningioma, glioma, sarcoma; Gynecology: Uterus (endometrial cancer), Cervix (cervical cancer, precancerous cervical dysplasia), Ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma)). Unclassified carcinoma, granulosa cell carcinoma, Sertoli cell carcinoma, germ cell carcinoma, malignant teratoma, vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, cystoid sarcoma (embryonic rhabdomyosarcoma), fallopian tube (carcinoma); hematology: blood (acute and chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus, lipoma, hemangioma, dermatofibroma, lupus erythematosus, psoriasis and adrenal glands: neuroblastoma.In some embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL).

[0142] In one embodiment, the cancer is a RAS family-related cancer, such as KRAS, NRAS, or HRAS-related cancer. In some embodiments, RAS family-related cancers are non-small cell lung cancer or pancreatic cancer. In one embodiment, the cancer is an SOS1-related cancer. In some embodiments, SOS1-related cancers are lung adenocarcinoma, embryonal rhabdomyosarcoma, Sertoli cell testis tumor, and granulosa cell tumor of the skin. In one embodiment, the cancer is an NF-1-related cancer.

[0143] Preferably, the cancer is pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, kidney cancer, gastric cancer, or bile duct cancer.

[0144] Terminology Explanation

[0145] Unless otherwise stated, some terms used in this specification and claims are defined as follows:

[0146] As used herein, "KRAS G12C" refers to a mutant form of the mammalian KRAS protein, which contains an amino acid at position 12 where glycine is replaced by cysteine.

[0147] The determination of the amino acid codons and residue positions of human KRAS is based on the amino acid sequence of UniProtKB / SwissProt P01116: variant p.Gly 12 Cys.

[0148] "KRAS G12D" refers to a mutant of the mammalian KRAS protein, containing an amino acid in which aspartic acid replaces glycine at position 12. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence determined by UniProtKB / Swi-protP01116: variant P.Gly 12 Asp.

[0149] "KRAS G12V" refers to a mutant of the mammalian KRAS protein, containing an amino acid in which valine replaces glycine at position 12. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified in UniProtKB / Swi-protP01116: variant P.Gly 12 Val.

[0150] "KRAS G12S" refers to a mutant of the mammalian KRAS protein, which contains an amino acid at position 12 where serine replaces glycine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01116: variant p.Gly 12Ser.

[0151] As used herein, "KRAS G12A" refers to a mutant form of the mammalian KRAS protein containing an amino acid at position 12 where alanine replaces glycine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01116: variant p.Gly 12Ala.

[0152] "KRAS G13D" refers to a mutant of the mammalian KRAS protein containing an amino acid at position 13 where aspartic acid replaces glycine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence determined by UniProtKB / SwissProt P01116: variant p.gly 13Asp.

[0153] "KRAS G13C" refers to a mutant of the mammalian KRAS protein, containing an amino acid at position 13 where glycine is replaced by cysteine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence determined by UniProtKB / SwissProt-Prot P01116: variant P.Gly 13Cys.

[0154] As used herein, "KRAS Q61L" refers to a mutant form of the mammalian KRAS protein containing a leucine residue at amino acid position 61, replacing the amino acid glutamine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence of UniProtKB / SwissProt P 01116: variant p.Gln61Leu.

[0155] As used herein, "KRAS A146T" refers to a mutant form of the mammalian KRAS protein containing an amino acid at position 146 where threonine replaces alanine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01116: variant p.al46Thr.

[0156] As used herein, "KRAS A146V" refers to a mutant form of the mammalian KRAS protein containing an amino acid at position 146 where valine replaces alanine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01116: variant p.al46Val.

[0157] As used herein, "KRAS A146P" refers to a mutant form of the mammalian KRAS protein containing a proline residue at amino acid position 146, replacing alanine. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01116: variant p.Al46Pro.

[0158] The term "HRAS G12C" as used herein refers to a mutant form of the mammalian HRAS protein containing an amino acid at position 12 where glycine is replaced by cysteine. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01112: variant p.Gly 12 Cys.

[0159] "HRAS G12D" refers to a mutant of a mammalian HRAS protein containing an amino acid substitution at position 12 where aspartic acid replaces glycine. The amino acid codons and residue positions of human HRAs are determined based on the amino acid sequence of UniProtKB / SwissProt P01112: variant P.Gly 12 Asp.

[0160] "HRAS G12S" refers to a mutant of a mammalian HRAS protein containing 12 amino acids in which glycine is replaced by serine. The amino acid codons and residue positions of human HRAS were determined based on the amino acid sequence determined by UniProtKB / SwissProtP01112: variant P.Gly 12Ser.

[0161] "HRAS G12A" refers to a mutant of a mammalian HRAS protein in which glycine is replaced by alanine at position 12. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProtP01112: variant P.Gly 12Ala.

[0162] "HRAS G13D" refers to a mutant of a mammalian HRAS protein containing an amino acid in which aspartic acid replaces glycine at position 13. The amino acid codons and residue positions of human HRAS were determined based on the amino acid sequence determined by UniProtKB / Swi-protP01112: variant p.gly 13Asp.

[0163] "HRAS G13C" is a mutant of the mammalian HRAS protein containing an amino acid in which cysteine ​​replaces glycine at position 13. The amino acid codons and residue positions of human HRAS were determined based on the amino acid sequence of UniProtKB / SwissProt-Prot P01112: variant P.Gly 13Cys.

[0164] The term "HRAS Q61L" as used herein refers to a mutant form of the mammalian HRAS protein containing a leucine residue at amino acid position 61, replacing the amino acid glutamine. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence identified from the UniProtKB / SwissProt P 01112 variant p.Gln61Leu.

[0165] As used herein, "HRAS A146T" refers to a mutant form of the mammalian HRAS protein containing a threonine residue at amino acid position 146 that replaces alanine. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01112: variant p.Al46Thr.

[0166] As used herein, "HRAS A146V" refers to a mutant form of the mammalian HRAS protein containing a valine amino acid at position 146, replacing alanine. The amino acid codons and residue positions of the human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01112: variant p.a146Val.

[0167] As used herein, “HRAS A146P” refers to a mutant form of the mammalian HRAS protein containing a proline residue at amino acid position 146 instead of alanine. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01112: variant p.A146Pro.

[0168] "NRAS G12C" refers to a mutant form of the mammalian NRAS protein, containing an amino acid where cysteine ​​replaces glycine at position 12. The amino acid codons and residue positions of human NRAS are determined according to UniProtKB / SwissProt-Prot P01111: variant P.Gly 12Cys.

[0169] "NRAS G12D" refers to a mutant of the mammalian NRAS protein, which contains an amino acid in which aspartic acid replaces glycine at position 12. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence determined by UniProtKB / Swi-protP01111: variant P.Gly 12 Asp.

[0170] "NRAS G12S" refers to a mutant of the mammalian NRAS protein, which contains an amino acid where serine replaces glycine at position 12. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProt P01111: variant p.Gly 12Ser.

[0171] “NRAS G12A” refers to a mutant of the mammalian NRAS protein, which contains an amino acid where alanine replaces glycine at position 12. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence of UniProtKB / SwissProt P01111: variant p.Gly 12Ala.

[0172] The term "NRAS G13D" used here refers to a mutant form of the mammalian NRAS protein, containing an amino acid substitute where aspartic acid replaces glycine at amino acid position 13. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence determined by UniProtKB / SwissProt P 01111: variant p.Gly 13Asp.

[0173] "HNRas G13C" refers to a mutant form of the mammalian NRAS protein, which contains an amino acid where cysteine ​​replaces glycine at position 13. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence of the variant P.Gly 13Cys (UniProtKB / SwissProt P01111).

[0174] The term "HRAS Q61L" as used herein refers to a mutant form of a mammalian HRAS protein containing leucine replacing glutamine at amino acid position 61. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence identified from the UniProtKB / SwissProt P 01112 variant p.Gln61Leu.

[0175] As used herein, “NRAS A146T” refers to a mutant form of the mammalian NRAS protein containing an amino acid where a threonine residue replaces alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01111: variant p.A146Thr.

[0176] As used herein, "NRAS A146V" refers to a mutant form of the mammalian NRAS protein containing a valine amino acid that replaces alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01111: variant p.a146Val.

[0177] As used in this article, "NRAS A146P" refers to a mutant form of the mammalian NRAS protein containing an amino acid where proline replaces alanine at position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01111: variant p.A146Pro.

[0178] As used here, “RAS family member” or “RAS family” refers to KRAS, HRAS, NRAS and their activating mutants, including those at G12, G13, Q61 and A146.

[0179] As used here, 'RAS family-related diseases or disorders' refers to diseases or disorders that are associated with, mediated by, or have activated RAS mutations, such as RAS at the G12, G13, Q61, or A146 positions. Non-limiting examples of RAS family-related diseases or illnesses include cancers associated with KRAS, HRAS, or NRAS G12C, KRAS, HRAS, or NRAS G12D, KRAS, HRAS, or NRAS G12V, KRAS, HRAS, or NRAS G12S, KRAS, HRAS, or NRAS G12A, KRAS, HRAS, or NRAS G13D, KRAS, HRAS, or NRAS G13C, KRAS, HRAS, or NRAS Q61L, KRAS, HRAS, or NRAS A146T, KRAS, HRAS, or NRAS A146V, or KRAS, HRAS, or NRAS 146P.

[0180] As used herein, “SOS1-related disease or disorder” refers to a disease or condition that is associated with, mediated by, or has an activated SOS1 mutation. Examples of activated SOS1 mutations include SOS1N233S and SOS1N233Y mutations.

[0181] Here, "SOS1N233S" refers to a mutant form of the mammalian SOS1 protein containing a serine substitute for glutamine at amino acid position 233. The amino acid codons and residue positions of human SOS1 were determined based on the amino acid sequence identified in UniProtKB / SwissProtQ 07889: variant p.Gln233Ser.

[0182] Here, "SOS1N233Y" refers to a mutant form of the mammalian SOS1 protein, in which a tyrosine substitute replaces glutamine at position 233. The amino acid codons and residue positions of human SOS1 were determined based on the amino acid sequence identified in UniProtKB / SwissProt Q 07889: variant p.Gln233Tyr.

[0183] As used herein, “SOS1 inhibitor” refers to the compounds of the present invention, as described herein, represented by formula (I). These compounds negatively inhibit all or part of the interaction between SOS1 and RAS family mutants or SOS1 activating mutations, thereby reducing and / or regulating the nucleotide exchange activity of the RAS family member-SOS1 complex.

[0184] As used herein, "KRAS inhibitor" refers to inhibitors known in the art that target various KRAS mutations. Inhibitors can be used for KRAS G12C, KRAS G12D, and KRAS G12V, among others.

[0185] As used herein, "MEK signaling pathway" inhibitors refer to inhibitors known in the art that target RAF, MEK, and ERK1 / 2.

[0186] "NF-1 / NF-2 related diseases or disorders" refers to diseases or disorders caused by loss-of-function mutations in the neurocellulase (NF-1) gene or the neurocellulase 2 (NF-2) gene.

[0187] As used herein, “loss-of-function mutation” refers to any point mutation, splice site mutation, fusion, nonsense mutation (amino acid mutation to a stop codon), intraframe mutation, or frameshift mutation, including insertions and deletions, as well as the loss of homozygosity of a gene encoding a protein in a target cell or cancer cell, resulting in the partial or complete loss of the presence, activity, and / or function of the encoded protein.

[0188] "alkyl" refers to a saturated aliphatic hydrocarbon group comprising 1-20 carbon atoms, or 1-10 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms, or 1-2 carbon atoms of a saturated straight-chain or branched monovalent hydrocarbon group, wherein the alkyl group may be independently and optionally substituted by one or more substituents described in this invention. Further examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group may be optionally substituted or unsubstituted.

[0189] "Alkenyl" refers to a monovalent hydrocarbon group with 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, in which at least one C or C is sp. 2 The double bond, wherein the alkenyl group may be independently and optionally substituted by one or more substituents described in this invention, specific examples of which include, but are not limited to, vinyl, allyl, and olefinic groups. The alkenyl group may be optionally substituted or unsubstituted.

[0190] "Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring comprises 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc.; polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups. The cycloalkyl group may be optionally substituted or unsubstituted.

[0191] "Spirocycloalkyl" refers to a polycyclic aromatic system with 5 to 18 quintiles, two or more cyclic structures, where the monocyclic rings share a carbon atom (called a spiro atom) with each other, and containing one or more double bonds within the rings, but none of the rings has fully conjugated π electrons. Preferably, it is 6 to 14 quintiles, more preferably 7 to 10 quintiles. Based on the number of shared spiro atoms between the rings, spirocycloalkyl is classified into monospiro, bispiro, or polyspirocycloalkyl, preferably monospiro and bispirocycloalkyl, and preferably 4 / 5, 4 / 6, 5 / 5, or 5 / 6 quintiles. Non-limiting examples of "spirocycloalkyl" include, but are not limited to:

[0192]

[0193] "Fused cycloalkyl" refers to a 5- to 18-membered polycyclic aromatic group containing two or more ring structures sharing a pair of carbon atoms. One or more rings may contain one or more double bonds, but none of the rings has fully conjugated π electrons. It is preferably a 6- to 12-membered aromatic system, more preferably a 7- to 10-membered system. Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to:

[0194]

[0195] "Bridged cycloalkyl" refers to an aromatic system consisting of 5 to 18 quintiles, containing two or more cyclic structures sharing two non-directly bonded carbon atoms, and wherein one or more rings may contain one or more double bonds, but none of the rings possesses fully conjugated π electrons. Preferably, it is a 6 to 12 quintile, more preferably a 7 to 10 quintile. Depending on the number of constituent rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to:

[0196]

[0197] The cycloalkyl ring can be fused to an aryl, heteroaryl, or heterocyclic ring, wherein the ring connected to the parent structure is a cycloalkyl ring, and non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc.

[0198] The terms “heterocyclic group,” “heterocyclic,” or “heterocyclic” are used interchangeably in this application. They all refer to a non-aromatic heterocyclic group comprising 3-12 saturated or partially unsaturated monocyclic, bicyclic, or tricyclic rings, wherein at least one ring atom is a heteroatom, such as oxygen, nitrogen, or sulfur. Preferably, it has a 5- to 7-membered monocyclic or a 7- to 10-membered bicyclic or tricyclic ring, which may contain 1, 2, or 3 atoms selected from nitrogen, oxygen, and / or sulfur. Examples of “heterocyclic group” include, but are not limited to, morpholino, oxobutyryl, thiomorpholino, tetrahydropyrano, 1,1-dioxo-thiomorpholino, piperidinyl, 2-oxo-piperidinyl, pyrrolyl, 2-oxo-pyrrolyl, piperazine-2-one, 8-oxa-3-aza-bicyclic [3.2.1]octyl, and piperazine. The heterocyclic ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group. The heterocyclic group may be optionally substituted or unsubstituted.

[0199] "Spirocycloheterocyclic group" refers to a polycyclic group with 5 to 18 members, two or more ring structures, in which the monocyclic rings share an atom with each other, and contains one or more double bonds within the rings, but none of the rings has fully conjugated π electrons. One or more ring atoms are selected from nitrogen, oxygen, sulfur, or S(O). m The heteroatom is a carbon atom, and the remaining ring atoms are carbon, m = 1 or 2. Preferably, it is 6 to 14 ternary, more preferably 7 to 10 ternary. Spirocyclic groups are classified into monospirocyclic, bispirocyclic, or multispirocyclic groups based on the number of shared spiro atoms between rings, preferably monospirocyclic and bispirocyclic groups. More preferably, it is a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospirocyclic group. Non-limiting embodiments of "spirocyclic group" include, but are not limited to:

[0200]

[0201] "Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more ring structures that share a pair of atoms with each other. One or more rings may contain one or more double bonds, but none of the rings have fully conjugated π electrons. One or more ring atoms are selected from nitrogen, oxygen, sulfur, or S(O). mThe heteroatom is composed of carbon atoms, with m = 1 or 2. Preferably, it is 6 to 14-membered, more preferably 7 to 10-membered. Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic group, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Non-limiting embodiments of the "fused heterocyclic group" include, but are not limited to:

[0202]

[0203] "Bridged heterocyclic group" refers to a polycyclic group with 5 to 18 members, containing two or more ring structures that share two atoms that are not directly connected. One or more rings may contain one or more double bonds, but none of the rings have fully conjugated π electrons. One or more ring atoms are selected from nitrogen, oxygen, sulfur, or S(O). m The heteroatom is carbon, and the remaining ring atoms are carbon, m = 1 or 2. Preferably, it is 6 to 14-membered, more preferably 7 to 10-membered. Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic group, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting embodiments of the "bridged heterocyclic group" include, but are not limited to:

[0204]

[0205] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be connected together in a fused manner. The term "aryl" includes aromatic groups such as phenyl, naphthyl, and tetrahydronaphthyl. Preferably, aryl groups are C6-C. 10 Aryl, more preferably phenyl and naphthyl, most preferably phenyl. The aryl group may be substituted or unsubstituted. The "aryl" group may be fused with a heteroaryl, heterocyclic, or cycloalkyl group, wherein the aryl ring is attached to the parent structure. Non-limiting embodiments include, but are not limited to:

[0206]

[0207] "Heteroaryl" refers to an aromatic 5- to 6-membered monocyclic or 9- to 10-membered bicyclic ring, which may contain 1 to 4 atoms selected from nitrogen, oxygen, and / or sulfur. Examples of "heteroaryl" include, but are not limited to, furanyl, pyridinyl, 2-oxo-1,2-dihydropyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiopheneyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrroleyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzo[a]dioxacyclopentenyl, benzimidazolyl, indoleyl, isoyindolyl, 1,3-dioxo-isoindolyl, quinolinyl, indazoleyl, benzisothiazolyl, benzo[a]oxazolyl, and benzisothiazolyl. Heteroaryl groups may be optionally substituted or unsubstituted. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. Non-limiting embodiments include, but are not limited to:

[0208]

[0209] "Alkoxy" refers to an (alkyl-O-) group. Alkyl groups are defined in the relevant section of this document. C1-C6 alkoxy groups are preferred. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, etc.

[0210] "Halogenated alkyl" refers to an alkyl group having one or more halogen substituents, wherein the alkyl group has the meaning as described in this invention. Examples of halogenated alkyl groups include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, perfluoroethyl, 1,1-dichloroethyl, 1,2-dichloropropyl, etc.

[0211] "Hydroxy" refers to the -OH group.

[0212] "Halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine, chlorine and bromine being preferred.

[0213] "Amino" refers to -NH2.

[0214] “Cyano” refers to -CN.

[0215] "Nitro" refers to -NO2.

[0216] "Benzyl" refers to -CH2-phenyl.

[0217] "Carboxyl group" refers to -C(O)OH.

[0218] "Acetyl" refers to -C(O)CH3 or Ac.

[0219] "Carboxylic acid ester group" refers to -C(O)O (alkyl) or (cycloalkyl), where alkyl and cycloalkyl are defined as described above.

[0220] "Optional" means that the event described may but does not have to happen. For example, "AR may be substituted by one or more Rc" implies that the AR group may be substituted by one or more Rc or may not be substituted by Rc.

[0221] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0222] Unless otherwise specified, the terms "substitution" or "substituted" in this specification refer to the substitution of a group by one or more groups selected from the following: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocyclic alkoxy, cycloalkylthio, heterocyclic alkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylic acid ester, =O, -C(O)R b -OC(O)R b -NR b R b -C(O)NR b R b -NR b C(O)R b -S(O)NR b R b or -S(O)2NR b R b , where R b The definition is as stated in general formula (I).

[0223] As used herein, the terms “subject,” “individual,” or “patient” are used interchangeably to refer to any animal, including mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, primates, and humans. In some embodiments, the patient is a human. In some embodiments, the subject has experienced and / or exhibited at least one symptom of a disease or illness to be treated and / or prevented. In some embodiments, the subject has been identified or diagnosed with cancer having a KRAS G12 or G13 mutation (e.g., determined by an FDA-approved regulatory agency, such as an FDA-approved test or kit). In some embodiments, the subject has a tumor that is positive for a KRAS G12C mutation, KRAS G12D mutation, KRAS G12S mutation, KRAS G12V mutation, KRAS G12A mutation, KRAS G13D mutation, or KRAS G13C mutation (e.g., determined by a regulatory agency-approved test or kit). The study subjects may be patients with tumors that are positive for KRAS G12C, KRAS G12D, KRAS G12V, KRAS G12S, KRAS G12A, KRAS G13D, or KRAS G13C mutations (e.g., by an approved regulatory agency, such as the FDA, an analysis, or a kit). The subject may be someone whose tumor has a KRAS G12C, KRAS G12D, KRAS G12V, KRAS G12S, KRAS G12A, KRAS G13D, or KRAS G13C mutation (e.g., the tumor was identified by an FDA-approved regulatory agency, kit, or assay). In some embodiments, the subject is suspected of having a cancer related to the KRAS G12 or G13 gene. In some embodiments, the subject has a clinical record indicating that they have a tumor with a KRAS G12C mutation (and optionally, a clinical record indicating that the subject should be treated with any of the compositions provided herein).

[0224] The term “pediatric patient” as used here refers to a patient under the age of 16 at the time of diagnosis or treatment. The term “child” can also be subdivided into the following subcategories: newborn (from birth to the first month); infant (1 month to 2 years); child (2 to 12 years); adolescent (12 to 21 years (until, but not including, their 22nd birthday)). Berhman RE, Kliegman R, Arvin AM, Nelson WE. Nelson's Pediatrics Textbook, 15th ed. Philadelphia: WBSaunders, 1996; Rudolph AM, et al. Rudolph's Pediatrics, 21st ed. New York: McGrow-Hill, 2002; and Avery MD, 1st LR. Pediatric Medicine, 2nd ed. Baltimore: Williams & Wilkins; 1994.

[0225] As used herein, the “effective amount” of a compound refers to an amount sufficient to negatively regulate or inhibit SOS1 enzyme activity.

[0226] As used herein, a "therapeuticly effective dose" of a compound refers to an amount sufficient to improve or reduce symptoms in some way, stop or reverse disease progression, or negatively regulate or inhibit SOS1 activity. This dose can be used as a single dose or taken in a regimen to be effective.

[0227] As used here, “treatment” means any way of improving or otherwise altering a patient’s condition, disorder, or the symptoms or pathology of a disease.

[0228] As stated herein, “to improve the symptoms of a particular disease by using a particular compound or pharmaceutical composition” means any reduction, whether permanent or temporary, lasting or transient, that is attributable to or related to the use of the composition.

[0229] The definition and conventional use of stereochemistry in this invention are generally referenced in the following literature:

[0230] SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric or chiral centers, and therefore exist as different stereoisomers. All stereoisomers of the compounds of this invention, including, but not limited to, diastereomers, enantiomers, transisomers, and mixtures thereof, such as racemic mixtures, constitute a part of this invention. Diastereomers can be separated into individual diastereomers based on their physicochemical differences by methods such as chromatography, crystallization, distillation, or sublimation. Enantiomers can be separated to convert a mixture of chiral isomers into a mixture of diastereomers by reacting with a suitable optically active compound (e.g., a chiral auxiliary agent, such as a chiral alcohol or Mosher's acyl chloride), separating the diastereomers and converting individual diastereomers into their corresponding pure enantiomers. The intermediates and compounds of this invention can also exist in different tautomeric forms, and all such forms are included within the scope of this invention. Many organic compounds exist in optically active forms, meaning they are capable of rotating the plane of polarized light. In describing optically active compounds, the prefixes D, L, or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes d, l, or (+), (-) are used to name the symbol for the plane polarization rotation of the compound; (-) or l indicates that the compound is levorotatory, and the prefix (+) or d indicates that the compound is dextrorotatory. These stereoisomers have the same order of atomic or atomic groups connected to each other, but their stereostructures are different. Specific stereoisomers can be enantiomers, and mixtures of isomers are generally called enantiomer mixtures. A 50:50 enantiomer mixture is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereodirection in chemical reactions. The terms "racemic mixture" and "racemate" refer to a mixture of two equimolar enantiomers that lack optical activity.

[0231] "Tautomer" or "tautomer form" refers to isomers of structures with different energies that can interconvert through a low energy barrier. For example, proton tautomers (i.e., proton-transfer tautomers) include interconversions via proton transfer, such as isomerizations of keto-enol and imine-enamine forms. Valence tautomers include interconversions involving the recombination of bonding electrons. Unless otherwise indicated, the structural formulas described in this invention include all isomer forms (e.g., enantiomers, diastereomers, and geometric isomers): for example, R, S configurations containing an asymmetric center, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers. Therefore, individual stereochemical isomers of the compounds of this invention, or mixtures of their enantiomers, diastereomers, or geometric isomers, are within the scope of this invention.

[0232] "Pharmaceutical salts" refer to salts of the compounds of this invention that are safe and effective when used in humans or animals. Salts of the compounds can be obtained by adding sufficient amounts of base or acid in a pure solution or suitable inert dissolution. Pharmaceutically available base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, etc., and pharmaceutically available acid addition salts include inorganic acid salts and organic acid salts, including hydrochloric acid, hydrobromic acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, monohydrogen sulfate, acetic acid, maleic acid, malonic acid, succinic acid, benzoic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, and methanesulfonic acid, etc. (See Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977)).

[0233] This invention provides a novel SOS1 inhibitor. Experimental results show that this pyrimidopyridone derivative exhibits excellent SOS1 inhibitory activity, along with excellent safety and selectivity. It can be used to prepare drugs for treating cancer, especially pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer, and bile duct cancer. Attached Figure Description

[0234] Figure 1 This is a graph showing the effects of the compounds involved in this invention on the KRAS / ERK1 / 2 signal transduction pathway in K-562 cells.

[0235] Figure 2 This is a diagram illustrating the in vivo antitumor effects of the SOS1 inhibitor and MRTX849, as described in this invention, used alone or in combination, on a nude mouse model of KRASG12C mutant MIAPaCa-2 pancreatic xenograft tumor.

[0236] Figure 3This is a graph showing the effect of the SOS1 inhibitor and MRTX849, as described in this invention, alone or in combination, on the body weight of a nude mouse model of a KRASG12C mutant MIAPaCa-2 pancreatic xenograft tumor.

[0237] Figure 4 This is a diagram illustrating the in vivo antitumor effects of the SOS1 inhibitor and trametinib, as described in this invention, used alone or in combination, on a nude mouse model of KRASG12D mutant HPAF-II pancreatic xenograft tumors.

[0238] Figure 5 This is a graph showing the effect of the SOS1 inhibitor and trametinib, as described in this invention, alone or in combination, on the body weight of a nude mouse model of a KRASG12D mutant HPAF-II pancreatic xenograft tumor. Detailed Implementation

[0239] The method of the present invention will be described below through specific embodiments to make the technical solution of the present invention easier to understand and master, but the present invention is not limited thereto. In the following embodiments... 1 The 1H NMR spectra were obtained using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. 1 HNMR representation: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If the coupling constant is provided, the unit is Hz.

[0240] The mass spectrometry results were obtained using an LC / MS instrument, with ESI as the ionization method.

[0241] High-performance liquid chromatograph (HPLC) models: Agilent 1260, Thermo Fisher U3000; Column model: Waters xbrige C18 (4.6*150mm, 3.5μm); Mobile phase: A: ACN, B: Water (0.1% H3PO4); Flow rate: 1.0 mL / min; Gradient: 5% A for 1 min, increase to 20% A within 4 min, increase to 80% A within 8 min, 80% A for 2 min, back to 5% A within 0.1 min; Wavelength: 220 nm; Column oven: 35℃.

[0242] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The silica gel plates used in TLC are 0.2mm-0.3mm in diameter, and the plates used for TLC separation and purification are 0.4mm-0.5mm in diameter.

[0243] Column chromatography typically uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0244] In the following examples, unless otherwise specified, all temperatures are in Celsius; unless otherwise specified, all starting materials and reagents are commercially available or synthesized according to known methods; commercially available materials and reagents are used directly without further purification; unless otherwise specified, commercially available manufacturers include, but are not limited to, Sinopharm Group, Bailingwei Technology Co., Ltd., TCI (Shanghai) Chemical Industry Development Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., and Shanghai Mairui Chemical Technology Co., Ltd.

[0245] CD3OD: Deuterated methanol

[0246] CDCl3: Deuterated chloroform

[0247] DMSO-d6: Deuterated dimethyl sulfoxide

[0248] Pd2(dba)3: Tris(dibenzylacetone)dipalladium

[0249] Pd(dppf)Cl2: [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride

[0250] XantPhos: 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene

[0251] XPhos: 2-Dicyclohexylphosphine-2,4,6-triisopropylbiphenyl

[0252] HATU: 2-(7-benzotriazole oxide)-N,N,N′,N′-tetramethylurea hexafluorophosphate

[0253] TLC: Thin-layer chromatography

[0254] HPLC: High Performance Liquid Chromatography

[0255] purity:

[0256] &:and

[0257] A hydrogen atmosphere refers to a reaction vessel connected to a hydrogen balloon with a volume of approximately 1L.

[0258] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0259] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃-30℃.

[0260] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system used for column chromatography to purify the compounds, or the developing solvent system for TLC included: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane: ethyl acetate. The volume ratio of the solvent varied depending on the polarity of the compound and could also be adjusted by adding a small amount of acidic or basic reagents, such as acetic acid or triethylamine.

[0261] Preparation of intermediates

[0262] Intermediate 1

[0263] (S)-2-((tetrahydrofuran-3-yl)oxy)ethyl acetate IN-1

[0264]

[0265] Step 1 (S)-Tetrahydrofuran-3-yl-4-nitrobenzene ester IN-1b

[0266] Under nitrogen protection, (R)-3-hydroxytetrahydrofuran IN-1a (10.0 g, 0.113 mol) was dissolved in tetrahydrofuran (130 mL). Nitrobenzoic acid (18.9 g, 0.113 mol) and triphenylphosphine (35.6 g, 0.136 mol) were added at room temperature. The mixture was cooled to 0 °C, and a tetrahydrofuran solution of diisopropyl azodicarbonate (27.8 g, 0.137 mol) in 20 mL was added dropwise. After the addition was complete, the reaction was carried out at room temperature for 2 hours. TLC showed that the starting material had completely reacted. The reaction solution was extracted with water and ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the white solid title compound IN-1b (29.8 g, crude product, containing a small amount of triphenylphosphine).

[0267] Step 2 (S)-3-hydroxytetrahydrofuran IN-1c

[0268] Compound IN-1b (29.8 g, crude) was dissolved in ethanol (300 mL), and sodium carbonate (25.0 g, 0.236 mol) was added. The mixture was heated to 85 °C and reacted for 2 hours. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography to obtain the oily compound IN-1c (8.1 g, two-step yield 81%).

[0269] Step 3: (S)-2-((tetrahydrofuran-3-yl)oxy)tert-butyl acetate IN-1d

[0270] Compound IN-1c (3.0 g, 34.05 mmol) was dissolved in toluene (40 mL), and tetrabutylammonium bromide (3.0 g, 9.30 mmol) and sodium hydroxide aqueous solution (30 mL, 360 mmol, 12 mol / L) were added. Tert-butyl bromoacetate (9.9 g, 50.75 mmol) was added with stirring at room temperature. After the addition was complete, the mixture was stirred overnight at 20 °C. TLC showed that the reaction was complete. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-1d (5.5 g, 80% yield) in oil.

[0271] 1 H NMR (400MHz, CDCl3) δ4.22-4.25 (m, 1H), 3.97 (s, 2H), 3.80-3.95 (m, 4H), 1.96-2.09 (m, 2H), 1.48 (s, 9H).

[0272] Step 4 (S)-2-((tetrahydrofuran-3-yl)oxy)acetic acid IN-1e

[0273] Compound IN-1d (5.5 g, 27.19 mmol) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (30 mL) was added. The mixture was heated to 45 °C and reacted for 2 hours. TLC showed that the reaction was essentially complete. The reaction solution was concentrated (dichloromethane was added three times) to give a pale yellow oil, title compound IN-1e (13.5 g, crude product), which was used directly in the next step.

[0274] Step 5 (S)-2-((tetrahydrofuran-3-yl)oxy)ethyl acetate IN-1

[0275] Compound IN-1e (13.5 g, crude) was dissolved in ethanol (100 mL), and concentrated sulfuric acid (30 mL) was added at room temperature. The mixture was heated to 85 °C and reacted for 5 hours. The reaction solution was cooled to room temperature, poured into ice-salt water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-1 (2.1 g, 44% yield in two steps).

[0276] 1 HNMR (400MHz, CDCl3) δ4.29-4.18 (m, 3H), 4.08 (s, 2H), 3.96-3.79 (m, 4H), 2.09-1.96 (m, 2H), 1.29 (t, J=7.2Hz, 3H).

[0277] Intermediate 2

[0278] (R)-(2-(5-(1-aminoethyl)thiophen-3-yl)benzyl)(methyl)carbamate IN-2

[0279]

[0280] Step 1 1-(4-bromo-2-thiophene) ethyl ketone IN-2b

[0281] 2-Acetthiophene IN-2a (10.0 g, 79.26 mmol) and aluminum trichloride (23.14 g, 173.54 mmol) were dispersed in chloroform (60 mL), and bromine (13.48 g, 84.36 mmol) dissolved in carbon tetrachloride (90 mL) was slowly added dropwise. After the addition was complete, the reaction was carried out at room temperature for 4 hours, and the reaction of the starting material 2-acetthiophene was monitored by TLC until complete. The reaction solution was poured into ice water and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2b (12.0 g, 74% yield) as a yellow oil.

[0282] 1 H NMR (400MHz, CDCl3) δ7.58 (d, J=1.2Hz, 1H), 7.58 (d, J=1.2Hz, 1H), 2.54 (s, 3H).

[0283] Step 2: (R)-N-(1-(4-bromothiophene-2-yl)ethylidene)-2-methylpropane-2-sulfinamide IN-2c

[0284] Tetraethyl titanate (60 mL) was added to a round-bottom flask, followed by the sequential addition of compound IN-2b (6.0 g, 29.26 mmol) and (R)-(+)-tert-butylsulfinamide (9.58 g, 79.04 mmol) at room temperature. The mixture was slowly heated to 100 °C and reacted for 1 hour, with TLC monitoring to ensure complete reaction of the starting material. The reaction mixture was cooled to room temperature, poured into ice water, and extracted with ethyl acetate (50 mL × 3 times). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2c (8.6 g, 95% yield).

[0285] Step 3: (R)-N-((R)-1-(4-bromothiophene-2-yl)ethyl)-2-methylpropane-2-sulfinamide IN-2d

[0286] Compound IN-2c (6.4 g, 20.76 mmol) was dissolved in a mixed solvent of tetrahydrofuran (60 mL) and water (1.2 mL). The solution was cooled to -50 °C, and sodium borohydride (2.23 g, 58.95 mmol) was added in portions. After the addition was complete, the solution was slowly heated to room temperature and reacted for 2 hours. TLC analysis showed that the reaction proceeded completely. The reaction solution was diluted with ethyl acetate, washed twice with saturated brine, washed twice with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2d (5.0 g, 78% yield).

[0287] Step 4 (R)-1-(4-bromothiophene-2-yl)-1-ethylamine IN-2e

[0288] Compound IN-2d (5.0 g, 16.11 mmol) and concentrated hydrochloric acid (12 N, 0.5 mL) were added to tetrahydrofuran (50 mL). The mixture was heated to 80 °C and stirred for 1 hour. The reaction mixture was monitored by TLC until the starting material was completely reacted. The reaction solution was cooled to room temperature, and the pH was adjusted to 8 by adding saturated sodium bicarbonate aqueous solution. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound IN-2e (3.34 g, crude product), which was used directly in the next step.

[0289] Step 5 (R)-(1-(4-bromothiophen-2-yl)ethyl)tert-butyl carbamate IN-2f

[0290] Compound IN-2e (12.4 g, crude) and triethylamine (7.3 g, 72.14 mmol) were dissolved in dichloromethane (150 mL), and di-tert-butyl dicarbonate (15.7 g, 71.94 mmol) was added. The mixture was stirred overnight at room temperature, and the reaction proceeds were monitored by TLC to ensure complete reaction. The reaction solution was washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2f (15.0 g, 82% yield in two steps).

[0291] Step 6 (R)-(1-(4-(2-formylphenyl)thiophen-2-yl)ethyl)tert-butyl carbamate IN-2g

[0292] Under nitrogen protection, compound IN-2f (15.0 g, 48.98 mmol) and 2-formylphenylboronic acid (8.8 g, 58.78 mmol) were dissolved in 1,4-dioxane (100 mL) and water (10 mL). At room temperature, [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (600 mg, 0.735 mmol) and sodium carbonate (10.4 g, 98.12 mmol) were added. The mixture was heated to 90 °C and reacted for 3 hours. TLC analysis showed that the reactants were essentially complete. The reaction solution was cooled to room temperature, diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2g (7.9 g, 49% yield).

[0293] Step 7 (R)-(1-(4-(2-((methylamino)methyl)phenyl)thiophene-2-yl)ethyl)tert-butyl carbamate IN-2h

[0294] Compound IN-2 g (7.9 g, 23.84 mmol) was dissolved in methylamine solution (25 mL, 30% ethanol solution) and stirred overnight at room temperature. Sodium cyanoborohydride (3.0 g, 47.74 mmol) was added, and the mixture was stirred at room temperature for 2 hours. TLC analysis showed that the reaction proceeds were essentially complete. The reaction solution was quenched with water, extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound IN-2 h (7.5 g, crude product), which was used directly in the next step.

[0295] LC-MS: m / z = 347.2 [M+H] +

[0296] Step 8 (R)-(2-(5-(1-((tert-butoxycarbonyl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carbamate IN-2i

[0297] Compound IN-2h (7.5 g, crude) was dissolved in dichloromethane (100 mL), and N,N-diisopropylethylamine (4.2 g, 32.50 mmol) and benzyl chloroformate (4.4 g, 25.79 mmol) were added sequentially. The mixture was stirred at room temperature for 2 hours, and the reaction was confirmed to be complete by TLC. The reaction solution was washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2i (10.6 g, 92% yield in two steps).

[0298] Step 9 (R)-(2-(5-(1-aminoethyl)thiophen-3-yl)benzyl)(methyl)carbamate IN-2

[0299] Compound IN-2i (7.0 g, 14.56 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (3 mL) was added. The mixture was stirred at room temperature for 1 hour, and the reaction was confirmed by TLC to be complete. The reaction solution was diluted with dichloromethane, washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-2 (3.2 g, 58% yield).

[0300] 1 H NMR (400MHz, DMSO-d6) δ7.64-6.96 (m, 11H), 6.18 (br, 2H), 5.16-4.98 (m, 2H), 4.60-4.42 (m, 3H), 2.78 (s, 3H), 1.50 (s, 3H).

[0301] Intermediate 3

[0302] (R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethylamine IN-3

[0303]

[0304] Step 1 1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl-1-one IN-3b

[0305] 3-Bromo-5-nitrotrifluorotoluene IN-3a (2.0 g, 7.41 mmol), tributyl(1-ethoxyethylene)tin (3.5 g, 9.69 mmol), and Pd(PPh3)2Cl2 (520 mg, 0.74 mmol) were added sequentially to toluene (25 mL). The mixture was heated to 100 °C overnight under nitrogen protection, and TLC showed complete reaction. The reaction solution was cooled to room temperature, and hydrochloric acid (15 mL, 3N) was added. The mixture was stirred for 30 minutes, filtered through a diatomaceous earth filter, and the filtrate was separated. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-3b (1.25 g, 72% yield) as a yellow oil.

[0306] 1 H NMR (400MHz, CDCl3) δ8.94 (s, 1H), 8.68 (s, 1H), 8.53 (s, 1H), 2.75 (s, 3H).

[0307] Step 2: (R,Z)-2-methyl-N-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethylidene)propane-2-sulfinamide IN-3c

[0308] A mixture of compound IN-3b (1.25 g, 5.36 mmol), (R)-(+)-tert-butylsulfinamide (974 mg, 8.04 mmol), and tetraethyl titanate (10 mL, 47.70 mmol) was heated to 80 °C and reacted for 3 hours. TLC showed a small amount of starting material remaining. The reaction mixture was cooled to room temperature, poured into ice water (60 mL), extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-3c (1.01 g, 56% yield) as a yellow oil.

[0309] Step 3: (R)-2-methyl-N-((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)propane-2-sulfinamide IN-3d

[0310] Compound IN-3c (260 mg, 0.77 mmol) was dissolved in tetrahydrofuran (2.5 mL) and water (0.05 mL). The solution was cooled to -60 °C, and sodium borohydride (74 mg, 1.95 mmol) was added in portions. After the addition was complete, the solution was stirred at -60 °C for 1 hour. TLC showed that the reaction was complete. The reaction solution was quenched dropwise with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the white solid title compound IN-3d (150 mg, 58% yield).

[0311] 1 H NMR (400MHz, CDCl3) δ 8.43-8.42 (m, 2H), 7.95 (s, 1H), 4.75-4.69 (m, 1H), 3.55 (d, J = 4.4Hz, 1H), 1.61 (d, J = 6.8Hz, 3H), 1.25 (s, 9H).

[0312] Step 4 (R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethylamine IN-3

[0313] Compound IN-3d (164 mg, 0.48 mmol) was dissolved in tetrahydrofuran (3 mL), and concentrated hydrochloric acid (0.5 mL) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 1 hour, and TLC showed that the reaction was complete. The pH of the reaction solution was adjusted to 8 by adding saturated sodium carbonate aqueous solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the yellow oily compound IN-3 (105 mg, 92% yield).

[0314] LC-MS: m / z = 235.1 [M+H] +

[0315] 1H NMR (400MHz, DMSO-d6) δ 8.56 (s, 1H), 8.32 (s, 1H), 8.24 (s, 1H), 4.24 (q, J=6.8Hz, 1H), 2.22 (br, 2H), 1.30 (d, J=2.8Hz, 3H).

[0316] Intermediate 4

[0317] (R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl-1-amine IN-4

[0318]

[0319] Step 1 1-Bromo-3-difluoromethyl-2-fluorobenzene IN-4b

[0320] 2-Fluoro-3-bromobenzaldehyde IN-4a (10.0 g, 49.26 mmol) was dissolved in dichloromethane (200 mL), cooled to 0 °C, and diethylaminotrifluoride (15.9 g, 98.64 mmol) was slowly added dropwise. After the addition was complete, the mixture was slowly brought to room temperature and stirred for 1 hour. TLC showed that the starting material reacted completely. The reaction solution was quenched in a saturated aqueous sodium bicarbonate solution, extracted with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the yellow liquid title compound IN-4b (8.1 g, 73% yield).

[0321] 1 H NMR (400MHz, CDCl3) δ7.68 (t, J=7.2Hz, 1H), 7.55 (t, J=7.2Hz, 1H), 7.14 (t, J=8.0Hz, 1H), 6.89 (t, J=54.8Hz, 1H).

[0322] Step 2: 1-(3-(difluoromethyl)-2-fluorophenyl)ethyl-1-one IN-4c

[0323] Compound IN-4b (8.1 g, 36.0 mmol) was dissolved in 1,4-dioxane (80 mL). Triethylamine (9.1 g, 89.9 mmol) and tributyl(1-ethoxyethylene)tin (15.6 g, 43.2 mmol) were added sequentially at room temperature. The mixture was bubbled under nitrogen for 15 minutes, followed by the addition of bis(triphenylphosphine)palladium dichloride (250 mg, 0.36 mmol). The mixture was then purged with nitrogen several times, heated to 100 °C, and stirred for 1 hour. TLC showed complete reaction. The reaction solution was cooled to room temperature, and dilute hydrochloric acid (14.4 mL, 72.0 mmol, 5 M) was added. The mixture was stirred at room temperature for 1 hour, and TLC showed complete reaction of the starting material. The reaction solution was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product purified by silica gel column chromatography to obtain the yellow liquid title compound IN-4c (5.6 g, 83% yield).

[0324] 1 H NMR (400MHz, CDCl3) δ8.00 (t, J=7.2Hz, 1H), 7.79 (t, J=6.8Hz, 1H), 7.34 (t, J=7.6Hz, 1H), 6.94 (t, J=54.8Hz, 1H), 2.67 (d, J=5.2Hz, 3H).

[0325] Step 3: 1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl) ethyl-1-one IN-4d

[0326] Potassium nitrate (45.6 g, 0.45 mmol) was dissolved in concentrated sulfuric acid (100 mL), stirred at room temperature for 30 minutes, cooled to approximately 0 °C, and compound IN-4c (8.5 g, 45.18 mmol) was slowly added dropwise. The mixture was stirred at 0 °C for 5 minutes, and TLC showed that the reaction was complete. The reaction solution was slowly added to ice water, extracted with ethyl acetate, and the organic phases were combined. The mixture was washed with saturated sodium bicarbonate solution and saturated brine, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-4d (9.0 g, 86% yield) as a pale yellow liquid.

[0327] 1 H NMR (400MHz, CDCl3) δ 8.88-8.86 (m, 1H), 8.66-8.64 (m, 1H), 7.12-6.85 (m, 1H), 2.73 (d, J=4.8Hz, 3H).

[0328] Step 4 (R,Z)-N-(1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethylene)-2-methylpropane-2-sulfinamide IN-4e

[0329] Compound IN-4d (9.0 g, 38.60 mmol) was dispersed in tetraethyl titanate (90 mL), and (R)-(+)-tert-butylsulfinamide (7.0 g, 57.76 mmol) was added at room temperature. The mixture was slowly heated to 100 °C and stirred for 1 hour. TLC showed that the starting material reacted completely. The reaction mixture was cooled to room temperature and poured into ice water with vigorous stirring. The mixture was stirred for 10 minutes, filtered, and the filter cake was repeatedly washed with ethyl acetate. The filtrate was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow oily compound, title compound IN-4e (9.0 g, crude), which was used directly in the next step.

[0330] LC-MS: m / z = 337.1 [M+H] +

[0331] Step 5 (R)-N-((R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl)-2-methylpropane-2-sulfinamide IN-4f

[0332] Compound IN-4e (9.0 g, crude) was dissolved in tetrahydrofuran (150 mL) and water (2 mL). The solution was cooled to approximately -60 °C, and sodium borohydride (3.1 g, 81.95 mmol) was added in portions. After the addition was complete, the solution was slowly heated to room temperature and the reaction continued for 2 hours. TLC showed that the starting material was exhausted. The reaction solution was quenched in ice water, extracted with ethyl acetate, and the organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the brown oily title compound IN-4f (2.8 g, 21% yield in two steps).

[0333] LC-MS: m / z = 339.1 [M+H] +

[0334] Step 6 (R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl-1-amine IN-4

[0335] Compound IN-4f (2.8 g, 8.28 mmol) was dissolved in tetrahydrofuran (30 mL), and concentrated hydrochloric acid (1.4 mL, 16.52 mmol) was added. The mixture was stirred at room temperature for 1 hour, and TLC showed that the starting material was exhausted. The reaction solution was cooled to room temperature, and the pH was adjusted to alkaline by adding saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a brown liquid. The liquid was then allowed to solidify into a brown solid, namely, title compound IN-4 (1.8 g, 95% yield).

[0336] LC-MS: m / z = 235.1 [M+H] +

[0337] 1H NMR (400MHz, DMSO-d6) δ 8.76-8.74 (m, 1H), 8.41-8.39 (m, 1H), 7.52-7.25 (m, 1H), 4.42 (q, J=6.4Hz, 1H), 2.48-2.24 (m, 2H), 1.35 (d, J=6.8Hz, 3H).

[0338] Intermediate 5

[0339] (R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethylamine IN-5

[0340]

[0341] Step 1 (R,Z)-N-(1-(3-(difluoromethyl)-2-fluorophenyl)ethylene)-2-methylpropane-2-sulfinamide IN-5a

[0342] Compound IN-4c (5.6 g, 29.76 mmol) was dissolved in tetraethyl titanate (50 mL), and (R)-(+)-tert-butylsulfinamide (10.8 g, 89.11 mmol) was added at room temperature. The mixture was slowly heated to 100 °C and stirred for 1 hour. TLC showed that the starting material reacted completely. The reaction solution was cooled to room temperature and poured into ice water with vigorous stirring. The mixture was stirred for 10 minutes, filtered, and the filter cake was repeatedly washed with ethyl acetate. The filtrate was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow oily compound, title compound IN-5a (9.5 g, crude), which was used directly in the next step.

[0343] LC-MS: m / z = 292.1 [M+H] +

[0344] Step 2: (R)-N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide IN-5b

[0345] Compound IN-5a (9.5 g, crude) was dissolved in tetrahydrofuran (100 mL) and water (2 mL). The solution was cooled to approximately -60 °C, and sodium borohydride (3.7 mg, 97.80 mmol) was added in portions. After the addition was complete, the solution was slowly heated to room temperature and the reaction continued for 2 hours. TLC showed that the starting material was exhausted. The reaction solution was quenched in ice water, extracted with ethyl acetate, and the organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-5b (2.9 g, 33% yield in two steps).

[0346] LC-MS: m / z = 294.1 [M+H] +

[0347] Step 3 (R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethylamine IN-5

[0348] Compound IN-5b (2.9 g, 9.88 mmol) was dissolved in tetrahydrofuran (50 mL), and concentrated hydrochloric acid (720 mg, 19.8 mmol) was added at room temperature. The mixture was heated to 80 °C and stirred for 1 hour. TLC showed that the starting material was exhausted. The reaction solution was cooled to room temperature, and the pH was adjusted to alkaline by adding saturated sodium bicarbonate solution. The solution was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a brown liquid, namely the title compound IN-5 (2.0 g, 107% yield).

[0349] LC-MS: m / z = 190.1 [M+H] +

[0350] 1 H NMR (400MHz, CDCl3) δ7.58 (t, J=7.4Hz, 1H), 7.47 (t, J=6.8Hz, 1H), 7.23 (t, J=7.6Hz, 1 H), 6.90 (t, J=55.2Hz, 1H), 4.45 (q, J=6.8Hz, 1H), 1.80 (s, 2H), 1.43 (d, J=6.4Hz, 3H).

[0351] Intermediate 6

[0352] Benzyl(S)-3-(2-ethoxy-2-oxoethoxy)pyrrolidine-1-carboxylic acid tert-butyl ester IN-6

[0353]

[0354] Step 1 (S)-3-hydroxypyrrolidine-1-carboxylic acid benzyl ester IN-6b

[0355] (S)-1-N-tert-Butoxycarbonyl-3-hydroxypyrrolidine IN-6a (5.0 g, 26.70 mmol) was dissolved in hydrochloric acid-methanol solution (40 mL, 4N) and reacted overnight at room temperature. The reaction solution was concentrated, and tetrahydrofuran (50 mL) and water (50 mL) were added. Sodium carbonate (14.1 g, 0.13 mmol) and benzyl chloroformate (6.8 g, 40.02 mmol) were added, and the mixture was stirred overnight at room temperature. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-6b (5.9 g, 100% yield) as a pale yellow oil.

[0356] Step 2: (S)-3-(2-(tert-butoxy)-2-oxoethoxy)pyrrolidine-1-carboxylic acid benzyl ester IN-6c

[0357] Compound IN-6b (5.9 g, 26.67 mmol) was dissolved in toluene (40 mL), and tetrabutylammonium bromide (2.7 g, 8.41 mmol) and sodium hydroxide aqueous solution (24 mL, 0.28 mol, 12 M) were added. Tert-butyl bromoacetate (8.2 g, 42.08 mmol) was added with stirring at room temperature. After the addition was complete, the mixture was stirred overnight at room temperature, and TLC showed that the reaction was complete. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the pale yellow oily title compound IN-6c (10.7 g, crude product), which was used directly in the next step.

[0358] Step 3: (S)-2-((1-((benzyloxy)carbonyl)pyrrolidine-3-yl)oxy)acetic acid IN-6d

[0359] Compound IN-6c (10.7 g, crude) was dissolved in dichloromethane (100 mL), and trifluoroacetic acid (10 mL) was added. The mixture was reacted overnight at room temperature, and TLC showed that the reaction was essentially complete. The reaction solution was concentrated (dichloromethane was added three times) to give a pale yellow oily compound IN-6d (10.1 g, crude), which was used directly in the next step.

[0360] Step 4: (S)-3-(2-ethoxy-2-oxoethoxy)pyrrolidine-1-carboxylic acid benzyl ester IN-6

[0361] Compound IN-6d (10.1 g, crude) was dissolved in N,N-dimethylformamide (50 mL), and potassium carbonate (23.2 g, 0.17 mol) and iodoethane (11.9 g, 84.06 mmol) were added. The reaction mixture was reacted at room temperature for 5 hours. The reaction solution was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the yellow oily title compound IN-6 (2.0 g, 23% yield in three steps).

[0362] Intermediate 7

[0363] (R)-3-(2-ethoxy-2-oxoethoxy)pyrrolidine-1-carboxylic acid benzyl ester IN-7

[0364]

[0365] Step 1 (R)-3-hydroxypyrrolidine-1-carboxylic acid benzyl ester IN-7b

[0366] (R)-1-N-tert-Butoxycarbonyl-3-hydroxypyrrolidine IN-7a (10.0 g, 53.40 mmol) was dissolved in hydrochloric acid-methanol solution (80 mL, 4 N) and reacted overnight at room temperature. The reaction solution was concentrated, and tetrahydrofuran (100 mL) and water (100 mL) were added. Sodium carbonate (28.2 g, 0.26 mmol) and benzyl chloroformate (13.6 g, 80.04 mmol) were added, and the mixture was stirred overnight at room temperature. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-7b (9.2 g, 78% yield) as a pale yellow oil.

[0367] Step 2: (R)-3-(2-(tert-butoxy)-2-oxoethoxy)pyrrolidine-1-carboxylic acid benzyl ester IN-7c

[0368] Compound IN-7b (9.2 g, 41.58 mmol) was dissolved in toluene (60 mL), and tetrabutylammonium bromide (4.0 g, 12.47 mmol) and sodium hydroxide aqueous solution (35 mL, 0.41 mol, 12 M) were added. Tert-butyl bromoacetate (12.2 g, 62.37 mmol) was added with stirring at room temperature. After the addition was complete, the mixture was stirred overnight at room temperature, and TLC showed that the reaction was complete. The reaction solution was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-7c (12.1 g, 86% yield) as a pale yellow oil.

[0369] Step 3 (R)-2-((1-((benzyloxy)carbonyl)pyrrolidine-3-yl)oxy)acetic acid IN-7d

[0370] Compound IN-7c (6.5 g, 19.38 mmol) was dissolved in dichloromethane (70 mL), and trifluoroacetic acid (7 mL) was added. The mixture was reacted overnight at room temperature, and TLC showed that the reaction was essentially complete. The reaction solution was concentrated (dichloromethane was added three times) to give a pale yellow oily compound IN-7d (7.2 g, crude), which was used directly in the next step.

[0371] Step 4 (R)-3-(2-ethoxy-2-oxoethoxy)pyrrolidine-1-carboxylic acid benzyl ester IN-7

[0372] Compound IN-7d (7.2 g, crude) was dissolved in N,N-dimethylformamide (50 mL), and potassium carbonate (16.6 g, 0.12 mol) and iodoethane (6.0 g, 38.80 mmol) were added at room temperature. The reaction mixture was reacted for 5 hours at room temperature. The reaction solution was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the yellow oily title compound IN-7 (4.2 g, 71% yield in two steps).

[0373] Intermediate 8

[0374] (R)-1-(3-(trifluoromethyl)phenyl)ethyl-1-amine IN-8

[0375]

[0376] Step 1 (R,Z)-2-methyl-N-(1-(3-(trifluoromethyl)phenyl)ethylidene)propane-2-sulfinamide IN-8b

[0377] m-Trifluoromethylacetophenone IN-8a (8.0 g, 42.52 mmol) and (R)-(+)-tert-butylsulfinamide (9.3 g, 76.53 mmol) were dissolved in isopropyl titanate (60 mL). The mixture was heated to 80 °C and stirred for 3 hours. The reaction was monitored by TLC until complete. The reaction solution was cooled to room temperature and poured into ice water (100 mL). Ethyl acetate was added and stirred. The mixture was filtered to remove insoluble matter. The aqueous phase was extracted with ethyl acetate, and the organic phase was washed with saturated brine. The solution was dried over anhydrous sodium sulfate and concentrated to give the title compound IN-8b (14.0 g, crude product), which was used directly in the next step.

[0378] Step 2: (R)-2-methyl-N-((R)-1-(3-(trifluoromethyl)phenyl)ethyl)propane-2-sulfinamide IN-8c

[0379] Compound IN-8b (14.0 g, crude) was dissolved in tetrahydrofuran (100 mL) and water (5 mL), cooled to -60 °C, and sodium borohydride (2.9 g, 76.54 mmol) was added in portions. After the addition was complete, the mixture was allowed to rise naturally to room temperature and stirred for 2 hours. The reaction was confirmed to be complete by TLC. The reaction solution was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-8c (7.9 g, 63% yield in two steps).

[0380] Step 3 (R)-1-(3-(trifluoromethyl)phenyl)ethyl-1-amine IN-8

[0381] Compound IN-8c (1.1 g, 3.75 mmol) was dissolved in tetrahydrofuran (20 mL), and concentrated hydrochloric acid (0.6 mL, 12 N) was added dropwise. The mixture was stirred at room temperature for 2 hours, and the reaction was confirmed to be complete by TLC. The reaction solution was neutralized with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, washed with saturated brine of the organic phase, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound IN-8 (700 mg, 99% yield).

[0382] Example 1

[0383] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0384]

[0385] Step 1: 6-Chloro-N,2-Dimethyl-4-pyrimidinamine 1b

[0386] 4,6-Dichloro-2-methylpyrimidine 1a (20.0 g, 0.123 mol) was added in portions to a methylamine solution (100 mL, 30% ethanol solution), and the reaction was carried out at room temperature for 1 hour. TLC showed that the starting material had completely reacted. The reaction solution was concentrated to give a white solid, title compound 1b (28.0 g, crude product), which was used directly in the next step.

[0387] LC-MS: m / z = 158.1 [M+H] +

[0388] Step 2: 6-Chloro-5-iodo-N,2-methyl-4-pyrimidinamine 1c

[0389] Compound 1b (28.0 g, crude) was dissolved in acetic acid (150 mL), and NIS (33.0 g, 0.147 mol) was added in portions at room temperature. The mixture was heated to 70 °C and reacted for 6 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, adjusted to neutral with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phases were combined. The mixture was washed with saturated sodium bicarbonate aqueous solution, saturated sodium sulfite aqueous solution, and saturated brine. The solution was dried over anhydrous sodium sulfate and concentrated to give a white solid, title compound 1c (37.2 g, crude containing succinimide), which was used directly in the next step.

[0390] LC-MS: m / z = 284.0 [M+H] +

[0391] Step 3: 6-Chloro-N,2-Dimethyl-5-vinyl-4-pyrimidinamine 1d

[0392] Compound 1c (15.0 g, crude) was dissolved in ethylene glycol dimethyl ether (150 mL), and potassium vinyltrifluoroborate (10.6 g, 79.13 mmol), potassium phosphate (28.1 g, 0.132 mol), and Pd(dppf)Cl2 dichloromethane complex (400 mg, 0.49 mmol) were added. The mixture was purged with nitrogen three times and heated to 80 °C overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 1d (3.87 g, 40% yield in three steps).

[0393] LC-MS: m / z = 184.1 [M+H] +

[0394] Step 4: 4-Chloro-2-methyl-6-(methylamino)pyrimidine-5-carboxaldehyde 1e

[0395] Compound 1d (3.87 g, 21.07 mmol) was dissolved in acetone (40 mL) and water (10 mL). N-methylmorpholine oxide (4.94 g, 42.17 mmol) was added, and the mixture was stirred until dissolved at room temperature. Potassium osmium tetroxide aqueous solution (catalytic amount) was added dropwise, and the reaction was allowed to proceed overnight at room temperature. TLC showed that the starting material had completely reacted. Sodium periodate (10.2 g, 47.69 mmol) was then added, and the reaction was allowed to proceed for 1 hour at room temperature. TLC showed that the intermediate phase had completely reacted. The reaction solution was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 1e (3.7 g, 94% yield).

[0396] LC-MS: m / z = 186.1 [M+H] +

[0397] Step 5 (S)-4-chloro-2,8-dimethyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 1f

[0398] Under nitrogen protection, compound 1e (900 mg, 4.85 mmol) was dissolved in tetrahydrofuran (40 mL), and intermediate IN-1 (1.1 g, 6.31 mmol) was added. The mixture was cooled to -60 °C, and lithium diisopropylamino (6.0 mL, 12.0 mmol, 2.0 M) was added dropwise. After the addition was complete, the mixture was reacted at -60 °C for 1 hour. TLC showed that the starting material had completely reacted. The reaction solution was quenched with citric acid, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was obtained by silica gel column chromatography. The crude product was then slurried with a mixture (petroleum ether / ethyl acetate = 3 / 1), filtered, and the white solid title compound 1f (560 mg, yield 39%) was obtained.

[0399] LC-MS: m / z = 296.1 [M+H] +

[0400] Step 6: 1 g of 2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0401] Compound 1f (95 mg, 0.32 mmol) was dissolved in dimethyl sulfoxide (3 mL). Intermediate IN-3 (72 mg, 0.31 mmol) and N,N-diisopropylethylamine (125 mg, 0.97 mmol) were added at room temperature. The mixture was heated to 80 °C and reacted for 16 hours. TLC showed complete reaction of the starting material. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give 1 g (82 mg, 54% yield) of the title compound as a yellow solid.

[0402] LC-MS: m / z = 494.2 [M+H] +

[0403] Step 7: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0404] 1 g (82 mg, 0.17 mmol) of compound was dissolved in methanol (5 mL), and palladium / carbon (60 mg, 10%) and concentrated hydrochloric acid (3 drops) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth, the filtrate was concentrated, and the crude product was purified by prep-TLC to give a white solid, title compound 1 (45 mg, yield 58%).

[0405] LC-MS: m / z = 464.2 [M+H] +

[0406] 1H NMR (400MHz, CD3OD) δ7.54 (s, 1H), 6.97-6.93 (m, 2H), 6.79 (s, 1H), 5.55 (q, J=7.2Hz, 1H), 5.12-5.08 (m, 1H), 4.05- 3.96 (m, 3H), 3.92-3.87 (m, 1H), 3.71 (s, 3H), 2.43 (s, 3H), 2.34-2.16 (m, 2H), 1.60 (d, J=7.2Hz, 3H). (99.45% purity by HPLC)

[0407] Example 2

[0408] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 2

[0409]

[0410] Step 1: 6-Chloro-2-methyl-4-pyrimidinamine 2a

[0411] 4,6-Dichloro-2-methylpyrimidine 1a (15.2 g, 93.25 mmol) was added to an aqueous ammonia solution (150 mL, 25%), and the mixture was heated to 65 °C and reacted for 3 hours. TLC showed that the starting material had completely reacted. The reaction solution was concentrated to give a white solid, title compound 2a (11.9 g, crude product), which was used directly in the next step.

[0412] LC-MS: m / z = 144.1 [M+H] +

[0413] Step 2: 6-Chloro-5-iodo-2-methyl-4-pyrimidinamine 2b

[0414] Compound 2a (11.9 g, crude) was added to acetic acid (120 mL), and N-iodosuccinimide (18.64 g, 82.85 mmol) was added in portions at room temperature. The mixture was heated to 70 °C and reacted for 4 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, adjusted to neutral with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phases were combined. The mixture was washed with saturated sodium bicarbonate aqueous solution, sodium sulfite aqueous solution, and saturated brine. The solution was dried over anhydrous sodium sulfate and concentrated to give a white solid, title compound 2b (11.82 g, crude), which was used directly in the next step.

[0415] LC-MS: m / z = 270.0 [M+H] +

[0416] Step 3: 6-Chloro-2-methyl-5-vinyl-4-pyrimidinamine 2c

[0417] Compound 2b (18.0 g, crude) was dissolved in ethylene glycol dimethyl ether (180 mL). At room temperature, potassium vinyltrifluoroborate (17.89 g, 0.13 mol), potassium phosphate (35.9 g, 0.17 mol), and Pd(dppf)Cl₂ dichloromethane complex (1.64 g, 2.01 mmol) were added sequentially. The mixture was purged with nitrogen three times, and the temperature was raised to 80 °C for 3 hours. TLC showed that the starting material reacted completely. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 2c (6.1 g, 25% yield in three steps).

[0418] Step 4: 4-Amino-6-chloro-2-methylpyrimidine-5-carboxaldehyde 2d

[0419] Compound 2c (5.00 g, 29.48 mmol) was dissolved in acetone and water (120 mL / 30 mL), and sodium periodate (14.32 g, 66.95 mmol) and potassium osmium tetroxide aqueous solution (catalytic amount) were added. The reaction was carried out at room temperature for 2 hours, and TLC showed that the starting material reacted completely. The reaction solution was extracted with water and ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 2d (2.5 g, yield 49%).

[0420] LC-MS: m / z = 172.1 [M+H] +

[0421] Step 5: (S)-4-chloro-2-methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 2e

[0422] Under nitrogen protection, compound 2d (2.50 g, 14.57 mmol) was dissolved in tetrahydrofuran (500 mL). Intermediate IN-1 (3.30 g, 18.94 mmol) was added at room temperature. The mixture was cooled to -60 °C, and lithium diisopropylamino (22 mL, 43.86 mmol, 2 M) was added dropwise. After the addition was complete, the reaction was carried out at -60 °C for 1 hour, and then slowly raised to room temperature and reacted overnight. The reaction solution was quenched with citric acid, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 2e (1.0 g, 24% yield) as a pale yellow solid.

[0423] LC-MS: m / z = 282.1 [M+H] +

[0424] Step 6: 2-Methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 2f

[0425] Compound 2e (100 mg, 0.35 mmol) was dissolved in dimethyl sulfoxide (3 mL). Intermediate IN-3 (83 mg, 0.35 mmol) and N,N-diisopropylethylamine (160 mg, 1.24 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted overnight. TLC showed the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 2f (80 mg, 48% yield).

[0426] LC-MS: m / z = 480.1 [M+H] +

[0427] Step 7: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 2

[0428] Compound 2f (80 mg, 0.17 mmol) was dissolved in methanol (5 mL), and palladium / carbon (30 mg, 10%) and concentrated hydrochloric acid (1 drop) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 2 (51 mg, yield 67%).

[0429] LC-MS: m / z = 450.2 [M+H] +

[0430] 1 H NMR (400MHz, CD3OD) δ7.57 (s, 1H), 6.96 (s, 2H), 6.80 (s, 1H), 5.55 (q, J=7.2Hz, 1H), 5.12-5.06 (m, 1H), 4. 05-3.95 (m, 3H), 3.93-3.85 (m, 1H), 2.39 (s, 3H), 2.34-2.15 (m, 2H), 1.60 (d, J=7.2Hz, 3H). (98.80% purity by HPLC)

[0431] Example 3

[0432] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 3

[0433]

[0434] Step 1: 4-Chloro-6-((4-methoxybenzyl)amino)pyrimidine-5-carboxaldehyde 3b

[0435] 4,6-Dichloro-5-pyrimidinecarboxaldehyde 3a (15.00 g, 84.76 mmol) was dissolved in dichloromethane (200 mL), cooled to 0 °C, and N,N-diisopropylethylamine (10.90 g, 84.34 mmol) was added dropwise. After the addition was complete, p-methoxybenzylamine (7.54 g, 54.96 mmol) was added dropwise. The reaction was carried out at 0 °C for 2 hours. TLC showed that the starting material reacted completely. The reaction solution was concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow solid crude product. The crude product was slurried with methyl tert-butyl ether, filtered, and the filter cake was washed with methyl tert-butyl ether to obtain a pale yellow solid title compound 3b (12.1 g, yield 51%).

[0436] LC-MS: m / z = 278.1 [M+H] +

[0437] Step 2: (S)-4-chloro-8-(4-methoxybenzyl)-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 3c

[0438] Under nitrogen protection, compound 3b (2.00 g, 7.20 mmol) was dissolved in tetrahydrofuran (300 mL), and intermediate IN-1 (1.63 g, 9.36 mmol) was added at room temperature. The mixture was cooled to -10 °C, and lithium diisopropylamino (10.8 mL, 21.60 mmol, 2 M) was added dropwise. After the addition was complete, the mixture was reacted at -10 °C for 3 hours, then slowly restored to room temperature and reacted overnight. The reaction solution was quenched with citric acid, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 3c (900 mg, 32% yield) as a brownish-yellow solid.

[0439] LC-MS: m / z = 388.1 [M+H] +

[0440] Step 3: 8-(4-methoxybenzyl)-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 3d

[0441] Compound 3c (500 mg, 1.29 mmol) was dissolved in dimethyl sulfoxide (6 mL). Intermediate IN-3 (300 mg, 1.28 mmol) and N,N-diisopropylethylamine (579 mg, 4.48 mmol) were added sequentially at room temperature. The mixture was heated to 90 °C and reacted overnight. TLC showed the reaction was complete. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the yellow solid title compound 3d (580 mg, 77% yield).

[0442] LC-MS: m / z = 586.2 [M+H] +

[0443] Step 4: 4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 3e

[0444] Compound 3d (580 mg, 0.99 mmol) was dissolved in trifluoroacetic acid (10 mL) and heated to 75 °C for 2 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, and the pH was adjusted to alkaline by adding saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a gray-brown solid, title compound 3e (430 mg, crude product), which was used directly in the next step.

[0445] LCMS: m / z = 466.2 [M+H] +

[0446] Step 5: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0447] Compound 3e (80 mg, crude) was dissolved in ethanol (10 mL) and water (3 mL). Reduced iron powder (48 mg, 0.86 mmol) and ammonium chloride (45 mg, 0.84 mmol) were added at room temperature, and the mixture was heated to 90 °C for 3 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 3 (42 mg, 52% yield in two steps).

[0448] LC-MS: m / z = 436.2 [M+H] +

[0449] 1H NMR (400MHz, CD3OD) δ8.20 (s, 1H), 7.61 (s, 1H), 7.05-6.77 (m, 3H), 5.55-5.43 (m, 1H), 5. 17-5.09 (m, 1H), 4.07-3.85 (m, 4H), 2.37-2.16 (m, 2H), 1.68-1.55 (m, 3H). (98.04% purity by HPLC)

[0450] Example 4

[0451] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidine

[0452]

[0453] Step 1: 8-Methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 4a

[0454] Compound 3e(10) (0 mg, 0.21 mmol) was dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (89 mg, 0.64 mmol) and methyl iodide (61 mg, 0.43 mmol) were added. The mixture was reacted overnight at room temperature, and TLC showed that the starting material had completely reacted. The reaction solution was added with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 4a (91 mg, crude product), which was used directly in the next step.

[0455] LC-MS: m / z = 480.2 [M+H] +

[0456] Step 2: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0457] Compound 4a (91 mg, crude) was dissolved in ethanol (10 mL) and water (3 mL). Reduced iron powder (53 mg, 0.95 mmol) and ammonium chloride (51 mg, 0.95 mmol) were added at room temperature, and the mixture was heated to 90 °C for 3 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give the title compound 4 (42 mg, 45% yield in two steps) as a pale yellow solid.

[0458] LC-MS: m / z = 450.2 [M+H] +

[0459] 1 H NMR (400MHz, CD3OD) δ8.31-8.25 (m, 1H), 7.58 (s, 1H), 6.93 (s, 2H), 6.79 (s, 1H), 5.49 (q, J=6.8Hz, 1H), 5.17- 5.10 (m, 1H), 4.06-3.86 (m, 4H), 3.77-3.66 (m, 3H), 2.36-2.17 (m, 2H), 1.61 (d, J=6.8Hz, 3H). (99.14% purity by HPLC)

[0460] Example 5

[0461] 4-(((R)-1-(5-amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 5

[0462]

[0463] Step 1 4-(((R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl)amino)-8-(4-methoxybenzyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 5a

[0464] Compound 3c (120 mg, 0.31 mmol) was dissolved in dimethyl sulfoxide (3 mL). Intermediate IN-4 (73 mg, 0.31 mmol) and N,N-diisopropylethylamine (140 mg, 1.08 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted for 8 hours. TLC showed complete reaction of the starting material. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 5a (66 mg, 36% yield).

[0465] LC-MS: m / z = 586.3 [M+H] +

[0466] Step 2: 4-(((R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 5b

[0467] Compound 5a (66 mg, 0.11 mmol) was dissolved in trifluoroacetic acid (2 mL) and heated to 75 °C for 2 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, and the pH was adjusted to alkaline by adding saturated sodium bicarbonate aqueous solution. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give the title compound 5b (27 mg, 52% yield) as a gray-brown solid.

[0468] LC-MS: m / z = 466.2 [M+H] +

[0469] Step 3: 4-(((R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl)amino)-8-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyridino[2,3-d]pyrimidin-7(8H)-one 5c

[0470] Compound 5b (27 mg, 0.058 mmol) was dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (16 mg, 0.12 mmol) and methyl iodide (25 mg, 0.18 mmol) were added at room temperature. The mixture was heated to 50 °C and reacted for 1 hour. TLC showed that the starting material had completely reacted. The reaction solution was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 5c (30 mg, crude product), which was used directly in the next step.

[0471] LC-MS: m / z = 480.2 [M+H] +

[0472] Step 4: 4-(((R)-1-(5-amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0473] Compound 5c (30 mg, crude) was dissolved in ethanol (10 mL) and water (3 mL). Reduced iron powder (18 mg, 0.32 mmol) and ammonium chloride (17 mg, 0.32 mmol) were added at room temperature, and the mixture was heated to 90 °C for 3 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give the title compound 5 (12 mg, 46% yield in two steps) as a pale yellow solid.

[0474] LC-MS: m / z = 450.3 [M+H] +

[0475] 1 H NMR (400MHz, CD3OD) δ8.26 (s, 1H), 7.64 (s, 1H), 7.01-6.83 (m, 2H), 6.76-6.73 (m, 1H), 5.64 (q, J=7.2Hz, 1H), 5.19-5.1 3 (m, 1H), 4.07-3.98 (m, 3H), 3.94-3.88 (m, 1H), 3.71 (s, 3H), 2.38-2.18 (m, 2H), 1.63 (d, J=6.8Hz, 3H). (96.10% purity by HPLC)

[0476] Example 6

[0477] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one

[0478]

[0479] Step 1 (E)-3-(4-chloro-2-methyl-6-(methylamino)pyrimidin-5-yl)methyl acrylate 6a

[0480] Compound 1c (10.5 g, 37.04 mmol) was dissolved in N,N-dimethylformamide (100 mL). Triethylamine (8.9 g, 87.95 mmol) and palladium acetate (200 mg, 0.89 mmol) were added at room temperature. The mixture was purged with nitrogen three times. Methyl acrylate (3.95 g, 45.88 mmol) was added using a syringe. The mixture was heated to 100 °C and reacted overnight. TLC showed that the starting material reacted completely. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was slurried in a mixture (petroleum ether / ethyl acetate = 5 / 1), filtered, and the filter cake was washed and dried to give a light yellow solid, title compound 6a (6.0 g, crude product), which was used directly in the next step.

[0481] LC-MS: m / z = 242.1 [M+H] +

[0482] Step 2: 4-Methoxy-2,8-dimethylpyrido[2,3-d]pyrimidin-7(8H)-one 6b

[0483] Compound 6a (2.17 g, crude) was added to methanol (30 mL), cooled to 0 °C, and sodium hydride (754 mg, 18.85 mmol, 60%) was added in portions. After the addition was complete, the mixture was heated to 60 °C and reacted for 6 hours. LC-MS showed that the starting material reacted completely. The reaction solution was cooled to 0 °C, quenched dropwise with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a pale yellow solid, title compound 6b (1.80 g, crude), which was used directly in the next step.

[0484] LC-MS: m / z = 206.1 [M+H] +

[0485] Step 3: 6-Bromo-4-methoxy-2,8-dimethylpyridino[2,3-d]pyrimidin-7(8H)-one 6c

[0486] Compound 6b (1.8 g, crude) was dissolved in N,N-dimethylformamide (20 mL), and N-bromosuccinimide (1.72 g, 9.66 mmol) was added in portions at room temperature. After the addition was complete, the mixture was heated to 50 °C and reacted for 2 hours. TLC showed that the starting material reacted completely. The reaction solution was cooled to room temperature, water was added, and the mixture was stirred for 20 minutes. The mixture was filtered, and the filter cake was dissolved in dichloromethane, dried over anhydrous sodium sulfate, and concentrated to give a pale yellow solid, title compound 6c (2.2 g, crude), which was used directly in the next step.

[0487] Step 4: 6-Bromo-4-hydroxy-2,8-dimethylpyrido[2,3-d]pyrimidin-7(8H)-one 6d

[0488] Compound 6c (2.2 g, crude) was dissolved in hydrochloric acid / 1,4-dioxane solution (20 mL, 4 M), and the mixture was heated to 65 °C and reacted for 2 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, diluted with dichloromethane, stirred for 30 minutes, filtered, and the filter cake was washed with dichloromethane and dried to give a pale yellow solid, title compound 6d (1.78 g, crude), which was used directly in the next step.

[0489] LC-MS: m / z = 272.0 [M+H] +

[0490] Step 5: 6-Bromo-2,8-dimethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl 2,4,6-triisopropylbenzenesulfonate 6e

[0491] Compound 6d (1.78 g, crude) was suspended in dichloromethane (20 mL). Triethylamine (2.74 g, 27.08 mmol) and 4-dimethylaminopyridine (83 mg, 0.68 mmol) were added at room temperature. The mixture was cooled to 0 °C, and 2,4,6-triisopropylbenzenesulfonyl chloride (2.46 g, 8.12 mmol) was added in portions. After the addition was complete, the mixture was allowed to react at room temperature for 0.5 h. TLC showed that the starting material had completely reacted. The reaction solution was added to water, extracted with dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 6e (2.45 g, 59% yield in five steps).

[0492] LC-MS: m / z = 536.0 [M+H] +

[0493] Step 6 (R)-6-bromo-2,8-dimethyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 6f

[0494] Compound 6e (1.45 g, 2.70 mmol) was dissolved in dimethyl sulfoxide (20 mL). Intermediate IN-3 (600 mg, 2.56 mmol) and N,N-diisopropylethylamine (1.15 g, 8.90 mmol) were added at room temperature. The mixture was heated to 80 °C and reacted for 3 hours. TLC showed complete reaction of the starting material. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a pale yellow solid, title compound 6f (1.08 g, 86% yield).

[0495] LC-MS: m / z = 486.0 [M+H] +

[0496] Step 7: (R)-5-(2,8-dimethyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-tert-butyl carboxylate 6h

[0497] Under nitrogen protection, compound 6f (200 mg, 0.41 mmol) and 1-tert-butoxycarbonyl-3,6-dihydro-2H-pyridine-5-boronic acid pinacol ester 6 g (190 mg, 0.61 mmol) were dissolved in 1,4-dioxane (15 mL) and water (3 mL). Pd(dppf)Cl2 dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (130 mg, 1.23 mmol) were added at room temperature. The mixture was heated to 100 °C and stirred for 1 hour. The reaction was monitored by TLC until complete. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to obtain a yellow solid, title compound 6h (251 mg, crude), which was used directly in the next step.

[0498] LC-MS: m / z = 589.3 [M+H] +

[0499] Step 8: 3-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 6i

[0500] Compound 6h (251 mg, crude) was dissolved in ethyl acetate (10 mL), and palladium / carbon (100 mg, 10%) was added at room temperature. The mixture was heated to 50 °C overnight under a hydrogen atmosphere, and the reaction was confirmed to be complete by LCMS. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth filter, the filter cake was washed with ethyl acetate, and the filtrate was concentrated to give a pale yellow oily substance, title compound 6i (260 mg, crude), which was used directly in the next step.

[0501] LC-MS: m / z = 561.3 [M+H] +

[0502] Step 9: 3-(4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 6j

[0503] Compound 6i (260 mg, 0.43 mmol) was dissolved in tetrahydrofuran (5 mL), and acetic anhydride (66 mg, 0.65 mmol) and triethylamine (113 mg, 0.88 mmol) were added at room temperature. The mixture was heated to 30 °C and reacted overnight. The reaction was confirmed to be complete by TLC. The reaction solution was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a pale yellow solid, title compound 6j (270 mg, crude product), which was used directly in the next step.

[0504] LC-MS: m / z = 603.3 [M+H] +

[0505] Step 10: N-(3-((1R)-1-((2,8-dimethyl-7-oxo-6-(piperidin-3-yl)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 6k

[0506] Compound 6j (270 mg, crude) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 2 hours, and TLC showed that the starting material had completely reacted. The reaction solution was adjusted to alkalinity with saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 6k (200 mg, crude), which was used directly in the next step.

[0507] Step 11 N-(3-((1R)-1-((2,8-dimethyl-6-(1-methylpiperidin-3-yl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 6I

[0508] Compound 6k (200 mg, crude) was dissolved in methanol (5 mL), and an aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium / carbon (70 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was filtered, and the filtrate was concentrated to give a yellow oily compound, title compound 6I (190 mg, crude), which was used directly in the next step.

[0509] LC-MS: m / z = 517.3 [M+H] +

[0510] Step 12 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 6

[0511] Compound 6I (190 mg, crude) was dissolved in methanol (6 mL), and sodium hydroxide aqueous solution (2 mL, 8.0 mmol, 4 N) was added at room temperature. The mixture was heated to 80 °C and reacted overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 6 (85 mg, 44% yield in six steps).

[0512] LC-MS: m / z = 475.3 [M+H] +

[0513] 1 HNMR (400MHz, CD3OD) δ8.10 (s, 1H), 7.01-6.94 (m, 2H), 6.80 (s, 1H), 5.55 (q, J=7.2Hz, 1H), 3.68 (s, 3H), 3.29-3.15 (m, 2H), 3. 12-3.03 (m, 1H), 2.43 (s, 6H), 2.29-2.18 (m, 2H), 2.00-1.75 (m, 3H), 1.61 (d, J=7.2Hz, 3H), 1.58-1.51 (m, 1H). (99.91% purity byHPLC)

[0514] Example 7

[0515] 4-(((R)-1-(5-amino-2-fluoro-3-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one

[0516]

[0517] Step 1 2-Fluoro-5-nitro-3-(trifluoromethyl)benzaldehyde 7b

[0518] At 0 °C, 3 g (15.62 mmol) of 2-fluoro-3-trifluoromethylbenzaldehyde 7a was dissolved in 6 mL of concentrated sulfuric acid, and 2.3 mL (68%) of nitric acid was added dropwise. After the addition was complete, the mixture was slowly brought to room temperature and reacted for 5 hours. TLC monitoring showed that a small amount of starting material remained. Ice water was slowly added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 7b (2.2 g, 59% yield).

[0519] 1H NMR (400MHz, CDCl3) δ10.42 (s, 1H), 8.96-8.94 (m, 1H), 8.77-8.75 (m, 1H).

[0520] Step 2 (2-fluoro-5-nitro-3-(trifluoromethyl)phenyl)methanol 7c

[0521] Compound 7b (940 mg, 3.96 mmol) was dissolved in anhydrous ethanol (15 mL), cooled to 0 °C, and sodium borohydride (226 mg, 5.97 mmol) was added in portions. After the addition was complete, the reaction was continued at 0 °C for 35 minutes. TLC showed that the starting material was completely converted. The reaction solution was quenched dropwise with saturated ammonium chloride aqueous solution, concentrated to remove ethanol, extracted with ethyl acetate in the aqueous phase, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the yellow liquid title compound 7c (792 mg, yield 84%).

[0522] Step 3 (5-amino-2-fluoro-3-(trifluoromethyl)phenyl)methanol 7d

[0523] Compound 7c (792 mg, 3.31 mmol) was dissolved in anhydrous ethanol (18 mL), and palladium / carbon (400 mg, 10%) was added. The mixture was incubated overnight at room temperature under a hydrogen atmosphere. TLC showed complete conversion of the starting material. The reaction solution was filtered with diatomaceous earth as an aid, the filter cake was washed with ethanol, and the filtrate was concentrated to give the title compound 7d (656 mg, crude product), which was used directly in the next step.

[0524] LC-MS: m / z = 210.1 [M+H] +

[0525] Step 4 (4-fluoro-3-(hydroxymethyl)-5-(trifluoromethyl)phenyl)carbamate 7e

[0526] Compound 7d (731 mg, crude), potassium carbonate (724 mg, 5.24 mmol), and benzyl chloroformate (716 mg, 4.20 mmol) were added sequentially to tetrahydrofuran (20 mL). The reaction was carried out at room temperature for 3 hours, and TLC showed that the reaction was complete. The reaction solution was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 7e (1.06 g, 84% yield in two steps).

[0527] 1 H NMR (400MHz, CDCl3) δ7.64 (t, J=5.6Hz, 2H), 7.40-7.33 (m, 5H), 6.90 (s, 1H), 5.20 (s, 2H), 4.77 (s, 2H).

[0528] Step 5 (4-fluoro-3-formyl-5-(trifluoromethyl)phenyl)carbamate 7f

[0529] Compound 7e (1.06 g, 3.09 mmol) was dissolved in dichloromethane (20 mL), and manganese dioxide (2.7 g, 31.03 mmol) was added. The reaction was carried out overnight at room temperature, and TLC showed that the reaction was complete. The reaction solution was filtered with diatomaceous earth as an aid, the filter cake was washed with dichloromethane, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 7f (934 mg, yield 89%).

[0530] Step 6: 7g of (4-fluoro-3-(1-hydroxyethyl)-5-(trifluoromethyl)phenyl)carbamate

[0531] Compound 7f (825 mg, 2.42 mmol) was dissolved in anhydrous tetrahydrofuran (18 mL), cooled to 0 °C, and methyl magnesium bromide (2.5 mL, 7.5 mmol, 3 M tetrahydrofuran solution) was added dropwise. After the addition was complete, the mixture was allowed to rise to room temperature and reacted overnight. TLC showed that the starting material was completely converted. The reaction solution was quenched dropwise with saturated ammonium chloride aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a white solid, title compound 7 g (858 mg, 99% yield).

[0532] 1 H NMR (400MHz, CDCl3) δ7.67-7.65 (m, 2H), 7.41-7.33 (m, 5H), 6.74 (s, 1H), 5.21 (s, 3H), 1.96-1.95 (m, 1H), 1.51 (d, J=6.4Hz, 3H).

[0533] Step 7 (3-acetyl-4-fluoro-5-(trifluoromethyl)phenyl)carbamate benzyl ester 7h

[0534] 7 g (858 mg, 2.40 mmol) of compound was dissolved in chloroform (20 mL), and manganese dioxide (3.2 g, 36.78 mmol) was added. The reaction was carried out overnight at room temperature. TLC showed that the starting material was not completely converted. The mixture was heated to reflux for 6 hours, and TLC showed that the starting material was completely converted. The reaction solution was cooled to room temperature, filtered with diatomaceous earth as an aid, the filter cake was washed with dichloromethane, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 7h (680 mg, 80% yield).

[0535] 1H NMR (400MHz, CDCl3) δ8.17 (s, 1H), 7.83-7.81 (m, 1H), 7.40-7.36 (m, 5H), 6.86 (s, 1H), 5.22 (s, 2H), 2.67 (d, J=5.2Hz, 3H).

[0536] Step 8 (R, E)-(3-(1-((tert-butylsulfinyl)imino)ethyl)-4-fluoro-5-(trifluoromethyl)phenyl)carbamate 7i

[0537] Compound 7h (620 mg, 1.75 mmol), (R)-(+)-tert-butylsulfinamide (317 mg, 2.62 mmol), and tetraethyl titanate (995 mg, 4.36 mmol) were sequentially added to tetrahydrofuran (15 mL). The mixture was heated to 60 °C and reacted overnight. TLC showed complete conversion of the starting material. The reaction mixture was cooled to room temperature, quenched dropwise with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 7i (343 mg, 43% yield).

[0538] Step 9 (3-((R)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-4-fluoro-5-(trifluoromethyl)phenyl)carbamate 7j

[0539] Compound 7i (343 mg, 0.75 mmol) was dissolved in tetrahydrofuran (3 mL) and water (2 drops). The mixture was cooled to -70 °C, and sodium borohydride (114 mg, 3.01 mmol) was added in portions. After the addition was complete, the reaction was continued for 1 hour. TLC showed that the starting material was not completely converted, so sodium borohydride (114 mg, 3.01 mmol) was added again, and the temperature was gradually increased to -45 °C for 1 hour. TLC showed that the starting material was completely converted, and two new spots were formed (LCMS confirmed that these were two isomers in a ratio of approximately 3:1, with the desired configuration being the predominant one). The reaction mixture was quenched dropwise with saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 7j (122 mg, 35% yield).

[0540] LC-MS: m / z = 461.1 [M+H] +

[0541] Step 10 (R)-(3-(1-aminoethyl)-4-fluoro-5-(trifluoromethyl)phenyl)carbamate 7k

[0542] Compound 7j (122 mg, 0.26 mmol) was dissolved in tetrahydrofuran (1 mL), and concentrated hydrochloric acid (0.3 mL) was added dropwise at room temperature. After the addition was complete, the reaction was continued at room temperature for 40 minutes. TLC showed that the starting material was completely converted. The reaction solution was quenched dropwise with saturated sodium carbonate solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 7k (92 mg, 97% yield).

[0543] Step 11 (3-((R)-1-((2,8-dimethyl-7-oxo-6-(((S)-tetrahydrofuran-3-yl)oxy)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-4-fluoro-5-(trifluoromethyl)phenyl)carbamate 7I

[0544] Compound 1f (88 mg, 0.30 mmol) was dissolved in N-methylpyrrolidone (5 mL). Compound 7k (97 mg, 0.27 mmol) and sodium carbonate (86 mg, 0.81 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted for 5 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 7I (48 mg, 28% yield).

[0545] Step 12 4-(((R)-1-(5-amino-2-fluoro-3-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 7

[0546] Compound 7I (48 mg, 0.078 mmol) was dissolved in methanol (5 mL), and palladium / carbon (60 mg, 10%) was added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 7 (14 mg, yield 37%).

[0547] LC-MS: m / z = 482.2 [M+H] +

[0548] 1H NMR (400MHz, CD3OD) δ7.60 (s, 1H), 6.92-6.90 (m, 1H), 6.79-6.77 (m, 1H), 5.66 (q, J=7.2Hz, 1H), 5.15-5.11 (m, 1H), 4.0 7-3.98 (m, 3H), 3.93-3.88 (m, 1H), 3.70 (s, 3H), 2.38 (s, 3H), 2.36-2.19 (m, 2H), 1.61 (d, J=7.2Hz, 3H). (94.15% purity by HPLC)

[0549] Example 8

[0550] N-((R)-1-(4-((((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxo7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)acetamide8

[0551]

[0552] Step 1 ((R)-1-(2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)tert-butyl carbamate 8b

[0553] Compound 6f (300 mg, 0.62 mmol) was dissolved in toluene (30 mL). (R)-3-tert-butoxycarbonylaminopyrrolidine 8a (172 mg, 0.92 mmol), Pd2(dba)3 (56 mg, 0.061 mmol), 1,1′-binaphthyl-2,2′-bis(diphenylphosphine) (57 mg, 0.092 mmol), and sodium tert-butoxide (178 mg, 1.85 mmol) were added at room temperature. The mixture was purged with nitrogen three times and heated to 100 °C overnight. TLC showed complete reaction of the starting material. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 8b (185 mg, 51% yield) as a brown solid.

[0554] LC-MS: m / z = 592.3 [M+H] +

[0555] Step 2: 6-((R)-3-aminopyrrolidone-1-yl)-2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 8c

[0556] Compound 8b (172 mg, 0.29 mmol) was dissolved in a 4 M, 4 mL, hydrochloric acid-methanol solution and reacted at room temperature for 1 hour. TLC showed that the starting material had completely reacted. The reaction mixture was adjusted to alkalinity by adding saturated sodium bicarbonate solution dropwise, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a brown solid, title compound 8c (160 mg, crude product), which was used directly in the next step.

[0557] LC-MS: m / z = 492.2 [M+H] +

[0558] Step 3: N-((R)-1-(2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)acetamide 8d

[0559] Compound 8c (160 mg, crude) was dissolved in dichloromethane (5 mL), and triethylamine (98 mg, 0.97 mmol) and acetic anhydride (50 mg, 0.49 mmol) were added. The mixture was reacted at room temperature for 1 hour, and TLC showed that the starting material had completely reacted. The reaction solution was diluted with dichloromethane, washed with hydrochloric acid (1 N), washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was slurried with a mixture (dichloromethane / petroleum ether = 1 / 10), filtered, and the filter cake was washed and dried to give a yellow solid, title compound 8d (120 mg, crude), which was used directly in the next step.

[0560] LC-MS: m / z = 534.2 [M+H] +

[0561] Step 4: N-((R)-1-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxo7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)acetamide 8

[0562] Compound 8d (120 mg, crude) was dissolved in ethyl acetate (3 mL) and methanol (1 mL), and concentrated hydrochloric acid (3 drops) and palladium / carbon (80 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was filtered through diatomaceous earth, and the filtrate was adjusted to alkalinity by adding ammonia-methanol solution (1 mL), concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 8 (50 mg, 34% yield in three steps).

[0563] LC-MS: m / z = 504.2 [M+H] +

[0564] 1 H NMR (400MHz, CD3OD) δ6.99 (s, 1H), 6.97 (s, 1H), 6.95 (s, 1H), 6.79 (s, 1H), 5.53 (q, J=6.8Hz, 1H), 4.44-4.38 (m, 1H), 3.75-3.61 (m , 5H), 3.50-3.37 (m, 2H), 2.40 (s, 3H), 2.28-2.19 (m, 1H), 1.95 (s, 3H), 1.93-1.88 (m, 1H), 1.60 (d, J=7.2Hz, 3H). (98.27% purity by HPLC)

[0565] Example 9

[0566] N-((S)-1-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)acetamide 9

[0567]

[0568] Step 1 ((S)-1-(2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)tert-butyl carbamate 9b

[0569] Compound 6f (300 mg, 0.62 mmol) was dissolved in toluene (30 mL), and (S)-3-tert-butoxycarbonylaminopyrrolidine 9a (172 mg, 0.92 mmol), Pd2(dba)3 (56 mg, 0.061 mmol), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (57 mg, 0.092 mmol) and sodium tert-butoxide (178 mg, 1.85 mmol) were added. The mixture was purged with nitrogen three times and heated to 100 °C overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a brown solid title compound 9b (105 mg, 29% yield).

[0570] LC-MS: m / z = 592.3 [M+H] +

[0571] Step 2: 6-((S)-3-aminopyrrolidone-1-yl)-2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 9c

[0572] Compound 9b (105 mg, 0.18 mmol) was dissolved in a hydrochloric acid-methanol solution (3 mL, 4 M) and reacted at room temperature for 1 hour. TLC showed that the starting material had completely reacted. The reaction solution was adjusted to alkalinity by dropwise addition of saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a brown solid, title compound 9c (90 mg, crude product), which was used directly in the next step.

[0573] Step 3: N-((S)-1-(2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)acetamide 9d

[0574] Compound 9c (90 mg, crude) was dissolved in dichloromethane (3 mL), and triethylamine (98 mg, 0.97 mmol) and acetic anhydride (50 mg, 0.49 mmol) were added. The mixture was reacted at room temperature for 1 hour, and TLC showed that the starting material had completely reacted. The reaction solution was diluted with dichloromethane, washed with 1N hydrochloric acid, washed with saturated brine of the organic phase, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give the yellow solid title compound 9d (71 mg, 75% yield in two steps).

[0575] LC-MS: m / z = 534.2 [M+H] +

[0576] Step 4: N-((S)-1-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidine-3-yl)acetamide 9

[0577] Compound 9d (71 mg, 0.13 mmol) was dissolved in ethyl acetate (3 mL) and methanol (1 mL), and concentrated hydrochloric acid (3 drops) and palladium / carbon (80 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was filtered through diatomaceous earth, and the filtrate was adjusted to alkalinity by adding ammonia-methanol solution (1 mL), concentrated, and the crude product was purified by Prep-TLC to give a white solid title compound 9 (30 mg, yield 45%).

[0578] LC-MS: m / z = 504.2 [M+H] +

[0579] 1 H NMR (400MHz, CD3OD) δ6.99 (s, 1H), 6.97 (s, 1H), 6.95 (s, 1H), 6.79 (s, 1H), 5.53 (q, J=7.2Hz, 1H), 4.44-4.38 (m, 1H), 3.75-3.61 (m , 5H), 3.50-3.37 (m, 2H), 2.40 (s, 3H), 2.28-2.19 (m, 1H), 1.95 (s, 3H), 1.93-1.88 (m, 1H), 1.60 (d, J=6.8Hz, 3H). (98.94% purity by HPLC)

[0580] Example 10

[0581] (R)-4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidin-4-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 10

[0582]

[0583] Step 1 (E)-3-(4-chloro-6-((4-methoxybenzyl)amino)pyrimidin-5-yl)methyl acrylate 10a

[0584] Compound 3b (7.41 g, 26.68 mmol) was dissolved in tetrahydrofuran (100 mL), and methoxyformylmethylenetriphenylphosphine (11.60 g, 34.69 mol) was added at room temperature. The mixture was heated to 65 °C and reacted for 2 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, concentrated, and the crude product was purified by silica gel column chromatography to give a pale yellow oil, title compound 10a (8.61 g, 97% yield).

[0585] LC-MS: m / z = 334.1 [M+H] +

[0586] Step 2: 4-Methoxy-8-(4-Methoxybenzyl)pyrido[2,3-d]pyrimidin-7(8H)-one 10b

[0587] Compound 10a (8.61 g, 25.80 mmol) was dissolved in methanol (100 mL), cooled to 0 °C, and sodium hydride (2.27 g, 56.75 mmol, 60%) was added. The mixture was heated to 65 °C and reacted overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, quenched with water, stirred for 0.5 h, filtered, the filter cake was washed with water, dissolved in dichloromethane / methanol, dried over anhydrous sodium sulfate, and concentrated to give a white solid, title compound 10b (7.67 g, crude product), which was used directly in the next step.

[0588] Step 3: 4-Methoxypyrido[2,3-d]pyrimidin-7(8H)-one 10c

[0589] Compound 10b (7.67 g, crude) was dissolved in trifluoroacetic acid (60 mL), and the mixture was heated to 75 °C and reacted for 3 hours. TLC showed that the starting material had completely reacted. The reaction solution was concentrated, neutralized with saturated sodium bicarbonate aqueous solution, and then dichloromethane (the product is insoluble in dichloromethane) was added. The mixture was stirred for 30 minutes, filtered, washed with water and dichloromethane, and dried to obtain a white solid, title compound 10c (3.90 g, crude), which was used directly in the next step.

[0590] LC-MS: m / z = 178.1 [M+H] +

[0591] Step 4: 4-Methoxy-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one 10d

[0592] Compound 10c (3.90 g, crude) was dissolved in N,N-dimethylformamide (40 mL), and potassium carbonate (4.56 g, 32.99 mmol) and methyl iodoforme (6.25 g, 44.03 mmol) were added at room temperature. The mixture was heated to 50 °C and reacted for 1 hour. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and a solid precipitated. The solid was filtered, washed with water, dissolved in dichloromethane, dried over anhydrous sodium sulfate, and concentrated to give a white solid, title compound 10d (3.04 g, crude), which was used directly in the next step.

[0593] Step 5: 6-Bromo-4-hydroxy-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one 10e

[0594] Compound 10d (2.45 g, crude) was dissolved in N,N-dimethylformamide (25 mL), and N-bromosuccinimide (2.74 g, 15.39 mmol) was added at room temperature. The mixture was heated to 100 °C and reacted overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and a solid precipitated. The mixture was stirred at room temperature for 0.5 hours, filtered, and the filter cake was washed with water and dried to give a white solid mixture (the product and the undemethylated mixture). This mixture was dissolved in hydrochloric acid / 1,4-dioxane (25 mL, 4N), heated to 70 °C, and reacted for 2 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, diluted with methanol, stirred for 0.5 hours, filtered, and the filter cake was washed with methanol and dried to give a pale yellow solid, title compound 10e (2.07 g, crude), which was used directly in the next step.

[0595] Step 6: 10f of 6-bromo-4-chloro-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one

[0596] Compound 10e (2.07 g, crude) was suspended in 1,4-dioxane (60 mL). N,N-diisopropylethylamine (4.17 g, 32.27 mmol) and phosphorus oxychloride (4.95 g, 32.28 mmol) were added at room temperature. The mixture was heated to 80 °C and reacted for 4 hours. TLC showed complete reaction of the starting material. The reaction solution was cooled, and the pH was adjusted to alkaline by dropwise addition of saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to give a pale yellow solid, title compound 10f (1.5 g, 26% yield in five steps).

[0597] LC-MS: m / z = 274.0 [M+H] +

[0598] 1 H NMR (400MHz, CDCl3) δ8.87 (s, 1H), 8.46 (s, 1H), 3.87 (s, 3H).

[0599] Step 7: (R)-6-bromo-8-methyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 10g

[0600] Compound 10f (880 mg, 3.21 mmol) was dissolved in dimethyl sulfoxide (6 mL). N,N-diisopropylethylamine (1.24 g, 9.59 mmol) and intermediate IN-3 (600 mg, 2.56 mmol) were added sequentially at room temperature. The mixture was heated to 80 °C and reacted for 4 hours. TLC showed complete reaction of the starting material. The reaction solution was cooled to room temperature, water was added, and the mixture was stirred for 0.5 hours. The mixture was filtered, the filter cake was washed with water, dissolved in ethyl acetate, washed with hydrochloric acid (1 N), washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 10 g (1.17 g, 97% yield).

[0601] LCMS: m / z = 472.1 [M+H] +

[0602] Step 8 (R)-8-methyl-6-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 10i

[0603] 10 g (150 mg, 0.32 mmol) of compound 10 h (106 mg, 0.48 mmol) and pinacol 4-methylcyclohexyl-1-enylboronic acid 10 h (106 mg, 0.48 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2 mL). Pd(dppf)Cl2 dichloromethane complex (18 mg, 0.022 mmol) and sodium carbonate (148 mg, 1.40 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted for 1 h. The reaction was confirmed to be complete by TLC. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the brown oily compound 10i (120 mg, 77% yield).

[0604] LC-MS: m / z = 489.2 [M+H] +

[0605] Step 9 (R)-4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidin-4-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 14

[0606] Compound 10i (120 mg, 0.25 mmol) was dissolved in methanol (5 mL), and palladium / carbon (50 mg, 10%) was added. The mixture was heated to 30 °C for 2 hours under a hydrogen atmosphere, and the reaction was confirmed to be complete by LC-MS. The reaction solution was filtered through diatomaceous earth, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain a yellow solid. The crude product was dissolved in dilute hydrochloric acid (1 N), extracted with ethyl acetate, and the organic phase was discarded. The aqueous phase was adjusted to alkalinity with saturated sodium bicarbonate solution, extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by Prep-TLC to obtain a pale yellow solid, title compound 14 (31 mg, yield 27%).

[0607] LC-MS: m / z = 461.3 [M+H] +

[0608] 1 H NMR (400MHz, CD3OD) δ8.33 (s, 1H), 8.22 (s, 1H), 7.00-6.96 (m, 1H), 6.94 (s, 1H), 6.79 (s, 1H), 5.52 (q, J=7.2Hz, 1H), 3.72-3.68 (m, 3H), 3.65- 3.56 (m, 2H), 3.52-3.43 (m, 1H), 3.24-3.08 (m, 2H), 2.98-2.93 (m, 1H), 2.91 (s, 3H), 2.20-2.07 (m, 3H), 1.63 (d, J=6.8Hz, 3H). (97.21% purity by HPLC)

[0609] Example 11

[0610] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpyrrolidone-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one

[0611]

[0612] Step 1 (R)-3-(8-methyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyridinyl[2,3-d]pyrimidin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester 11b

[0613] 10 g (200 mg, 0.42 mmol) of compound 1 and 1-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-boronic acid pinacol ester 11a (187 mg, 0.63 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2 mL). At room temperature, a Pd(dppf)Cl2 dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (130 mg, 1.23 mmol) were added. The mixture was purged with nitrogen three times, heated to 100 °C, and reacted for 1 hour. The reaction was confirmed to be complete by TLC. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to give a yellow solid, title compound 11b (200 mg, 84% yield).

[0614] LC-MS: m / z = 561.3 [M+H] +

[0615] Step 2: (R)-3-(4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester 11c

[0616] Compound 11b (200 mg, 0.36 mmol) was dissolved in ethyl acetate (10 mL), and palladium / carbon (50 mg, 10%) was added at room temperature. The mixture was heated to 50 °C overnight under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth filter, the filter cake was washed with ethyl acetate, and the filtrate was concentrated to give a yellow solid, title compound 11c (170 mg, crude product), which was used directly in the next step.

[0617] LC-MS: m / z = 529.2 [M+H]-

[0618] Step 3: (R)-3-(4-((1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester 11d

[0619] Compound 11c (170 mg, crude) was dissolved in tetrahydrofuran (5 mL), and acetic anhydride (124 mg, 1.21 mmol) and N,N-diisopropylethylamine (235 mg, 1.82 mmol) were added at room temperature. The mixture was heated to 40 °C and reacted overnight. The reaction was confirmed to be complete by TLC. The reaction solution was cooled to room temperature, diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a brown solid, title compound 11d (207 mg, crude), which was used directly in the next step.

[0620] LC-MS: m / z = 573.3 [M+H] +

[0621] Step 4: 3-(4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidine-1-carboxylic acid tert-butyl ester 11e

[0622] Compound 11d (207 mg, crude) was dissolved in methanol (10 mL), and palladium / carbon (50 mg, 10%) was added. The mixture was heated to 30 °C and reacted for 2 hours under a hydrogen atmosphere. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth sieve, the filter cake was washed with methanol, and the filtrate was concentrated to give a yellow solid, title compound 11e (200 mg, crude), which was used directly in the next step.

[0623] Step 5: N-(3-((1R)-1-((8-methyl-7-oxo-6-(pyrrolidine-3-yl)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 11f

[0624] Compound 11e (200 mg, crude) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 2 hours, and TLC showed that the starting material had completely reacted. The reaction solution was adjusted to alkalinity with saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 11f (187 mg, crude), which was used directly in the next step.

[0625] Step 6: 11g of N-(3-((1R)-1-((8-methyl-6-(1-methylpyrrolidone-3-yl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide

[0626] Compound 11f (187 mg, crude) was dissolved in methanol (5 mL), and formaldehyde aqueous solution (100 mg, 1.23 mmol, 37%) and palladium / carbon (70 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was filtered through diatomaceous earth, the filter cake was washed with methanol, and the filtrate was concentrated to give a yellow oily substance, title compound 11 g (190 mg, crude), which was used directly in the next step.

[0627] Step 7: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpyrrolidone-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 11

[0628] Compound 11 g (190 mg, crude product) was dissolved in methanol (6 mL), and sodium hydroxide aqueous solution (3 mL, 4N) was added at room temperature. The mixture was heated to 70 °C and reacted overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to Prep-TLC to give a white solid, title compound 11 (60 mg, 40% yield in six steps).

[0629] LC-MS: m / z = 447.3 [M+H] +

[0630] 1 H NMR (400MHz, CD3OD) δ8.34 (s, 1H), 8.28 (s, 1H), 6.96 (s, 1H), 6.93 (s, 1H), 6.79 (s, 1H), 5.50 (q, J=6.8Hz, 1H), 3.79-3.71 (m, 1H), 3.70 (s, 3H), 3. 45-3.36(m, 2H), 3.29-3.24(m, 1H), 3.23-3.15(m, 1H), 2.80(s, 3H), 2.5 2-2.40 (m, 1H), 2.26-2.15 (m, 1H), 1.62 (d, J=7.2Hz, 3H). (94.63% purity by HPLC)

[0631] Example 12

[0632] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-((1-methylpyrrolidone-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 12

[0633]

[0634] The compound of Example 12 was synthesized according to the synthesis method of Example 13.

[0635] Example 13

[0636] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(((S)-1-methylpyrrolidine-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 13

[0637]

[0638] Step 1: 4-Chloro-6-(methylamino)pyrimidine-5-carboxaldehyde 13a

[0639] 4,6-Dichloro-5-pyrimidinecarboxaldehyde 3a (52.0 g, 0.29 mol) was dissolved in dichloromethane (500 mL), cooled to 0 °C, and triethylamine (59.5 g, 0.59 mol) was added dropwise. Methylamine hydrochloride (16.9 g, 0.25 mol) was added in portions. After the addition was complete, the reaction was carried out at 0 °C for 2 hours, and TLC showed that the reaction was complete. The reaction solution was filtered through diatomaceous earth, the filtrate was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography to obtain a crude product. The crude product was slurried with methyl tert-butyl ether, filtered, and the filter cake was washed with methyl tert-butyl ether to give a pale yellow solid, title compound 13a (23.5 g, yield 46%).

[0640] LCMS: m / z = 172.1 [M+H] +

[0641] Step 2: (S)-3-((4-chloro-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 13b

[0642] Under nitrogen protection, compound 13a (900 mg, 5.24 mmol) was dissolved in tetrahydrofuran (15 mL), and intermediate IN-6 (2.1 g, 6.81 mmol) was added. The mixture was cooled to -15 °C, and lithium diisopropylamino (6.6 mL, 13.20 mmol, 2.0 M) was added dropwise. After the addition was complete, the mixture was reacted at -10 °C for 5 hours, and then at room temperature overnight. TLC showed that the reaction was complete. The reaction solution was quenched dropwise with saturated citric acid aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the yellow oil title compound 13b (144 mg, yield 7%).

[0643] LCMS: m / z = 415.1 [M+H] +

[0644] Step 3: (S)-3-((8-methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 13c

[0645] Compound 13b (144 mg, 0.35 mmol) was dissolved in dimethyl sulfoxide (4 mL), and intermediate IN-3 (82 mg, 0.35 mmol) and N,N-diisopropylethylamine (135 mg, 1.04 mmol) were added. The reaction mixture was heated to 90 °C and reacted overnight. TLC showed that the starting material had completely reacted. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with 1N hydrochloric acid, then with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to Prep-TLC to give the title compound 13c (194 mg, 92% yield) as a yellow solid.

[0646] Step 4: (S)-3-((4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 13d

[0647] Compound 13c (194 mg, 0.32 mmol) was dissolved in ethanol (10 mL) and water (3 mL). Reduced iron powder (89 mg, 1.58 mmol) and ammonium chloride (86 mg, 1.58 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 5 hours. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth filter, and the filtrate was extracted with water and ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a brownish-yellow solid, title compound 13d (140 mg, 76% yield).

[0648] Step 5: (S)-3-((4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 13e

[0649] Compound 13d (140 mg, 0.24 mmol) was dissolved in tetrahydrofuran (5 mL), and acetic anhydride (47 mg, 0.47 mmol) and N,N-diisopropylethylamine (91 mg, 0.70 mmol) were added. The mixture was reacted overnight at room temperature, and the reaction was confirmed to be complete by TLC. The reaction solution was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 13e (104 mg, 69% yield) as a pale yellow solid.

[0650] Step 6: N-(3-((R)-1-((8-methyl-7-oxo-6-(((S)-pyrrolidine-3-yl)oxy)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 13f

[0651] Compound 13e (104 mg, 0.16 mmol) was dissolved in ethanol (5 mL), and palladium / carbon (60 mg, 10%) was added. The reaction was carried out at room temperature for 3 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through diatomaceous earth, the filter cake was washed with methanol, and the filtrate was concentrated to give a pale yellow solid, title compound 13f (90 mg, crude product), which was used directly in the next step.

[0652] Step 7: N-(3-((R)-1-((8-methyl-6-((((S)-1-methylpyrrolidine-3-yl)oxy)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 13g

[0653] Compound 13f (90 mg, crude) was dissolved in methanol (5 mL), and an aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium / carbon (70 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, and the filtrate was concentrated to give a pale yellow solid, title compound 13 g (95 mg, crude), which was used directly in the next step.

[0654] Step 8: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(((S)-1-methylpyrrolidine-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 13

[0655] Compound 13 g (95 mg, crude) was dissolved in methanol (4 mL), and sodium hydroxide aqueous solution (4 mL, 16.00 mmol, 4 N) was added at room temperature. The mixture was heated to 80 °C and reacted overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 13 (40 mg, 52% yield in three steps).

[0656] LCMS: m / z = 463.2 [M+H] +

[0657] 1H NMR (400MHz, CD3OD) δ8.29 (s, 1H), 7.53 (s, 1H), 6.93 (s, 2H), 6.79 (s, 1H), 5.55 (t, J=7.2Hz, 1H), 5.10-5.03 (m, 1H), 3.74 (s, 3H), 3 .20-3.13 (m, 1H), 3.01-2.89 (m, 2H), 2.79-2.71 (m, 1H), 2.51 (s, 3H), 2.48-2.38 (m, 1H), 2.17-2.08 (m, 1H), 1.62 (d, J=7.2Hz, 3H).

[0658] Example 14

[0659] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(((R)-1-methylpyrrolidine-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 14

[0660]

[0661] Step 1 (R)-3-((4-chloro-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 14a

[0662] Under nitrogen protection, compound 13a (700 mg, 4.08 mmol) was dissolved in tetrahydrofuran (40 mL), and intermediate IN-7 (1.6 g, 5.30 mmol) was added. The mixture was cooled to -15 °C, and lithium diisopropylamino (5.1 mL, 10.20 mmol, 2.0 M) was added dropwise. After the addition was complete, the mixture was reacted at -10 °C for 5 hours, and then at room temperature overnight. TLC showed that the reaction was complete. The reaction solution was quenched dropwise with saturated citric acid aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a yellow oily title compound 14a (176 mg, 10% yield).

[0663] LCMS: m / z = 415.2 [M+H] +

[0664] Step 2: (R)-3-((8-methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 14b

[0665] Compound 14a (176 mg, 0.42 mmol) was dissolved in dimethyl sulfoxide (4 mL). Intermediate IN-3 (99 mg, 0.42 mmol) and N,N-diisopropylethylamine (164 mg, 1.27 mmol) were added at room temperature. The mixture was heated to 90 °C and reacted overnight. TLC showed the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with 1N hydrochloric acid, saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a brown solid, title compound 14b (252 mg, 97% yield).

[0666] LCMS: m / z = 613.2[M+H] +

[0667] Step 3: (R)-3-((4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 14c

[0668] Compound 14b (252 mg, 0.41 mmol) was dissolved in ethanol (10 mL) and water (3 mL). Reduced iron powder (115 mg, 2.06 mmol) and ammonium chloride (111 mg, 2.06 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 5 hours. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth filter, and the filtrate was extracted with water and ethyl acetate. The combined phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a brownish-yellow solid, title compound 14c (125 mg, yield 52%).

[0669] LCMS: m / z = 583.3 [M+H] +

[0670] Step 4: (R)-3-((4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylic acid benzyl ester 14d

[0671] Compound 14c (110 mg, 0.19 mmol) was dissolved in tetrahydrofuran (5 mL), and acetic anhydride (38 mg, 0.38 mmol) and N,N-diisopropylethylamine (73 mg, 0.57 mmol) were added. The mixture was reacted overnight at room temperature, and the reaction was confirmed to be complete by TLC. The reaction solution was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 14d (80 mg, 67% yield) as a pale yellow solid.

[0672] Step 5: N-(3-((R)-1-((8-methyl-7-oxo-6-(((R)-pyrrolidine-3-yl)oxy)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 14e

[0673] Compound 14d (80 mg, 0.13 mmol) was dissolved in ethanol (5 mL), and palladium / carbon (40 mg, 10%) was added. The reaction was carried out at room temperature for 3 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, and the filtrate was concentrated to give a pale yellow solid, title compound 14e (85 mg, crude product), which was used directly in the next step.

[0674] Step 6: N-(3-((R)-1-((8-methyl-6-((((R)-1-methylpyrrolidine-3-yl)oxy)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 14f

[0675] Compound 14e (85 mg, crude) was dissolved in methanol (5 mL), and an aqueous formaldehyde solution (85 mg, 1.05 mmol, 37%) and palladium / carbon (30 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, and the filtrate was concentrated to give a pale yellow solid, title compound 14f (90 mg, crude), which was used directly in the next step.

[0676] Step 7: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(((R)-1-methylpyrrolidine-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 14

[0677] Compound 14f (90 mg, crude) was dissolved in methanol (4 mL), and an aqueous sodium hydroxide solution (4 mL, 16.00 mmol, 4 N) was added. The mixture was heated to 80 °C and reacted overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 14 (35 mg, 59% yield in three steps).

[0678] LCMS: m / z = 463.2 [M+H] +

[0679] 1H NMR (400MHz, CD3OD) δ8.29 (s, 1H), 7.55 (s, 1H), 6.93 (s, 2H), 6.79 (s, 1H), 5.50 (q, J=6.8Hz, 1H), 5.11-5.04 (m, 1H), 3.74 (s, 3H), 3 .23-3.16 (m, 1H), 3.06-2.94 (m, 2H), 2.83-2.74 (m, 1H), 2.54 (s, 3H), 2.50-2.38 (m, 1H), 2.17-2.07 (m, 1H), 1.62 (d, J=6.8Hz, 3H).

[0680] Example 15

[0681] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 15

[0682]

[0683] Step 1 (R)-5-(8-methyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester 15a

[0684] 10 g (240 mg, 0.51 mmol) of compound and 6 g (236 mg, 0.76 mmol) of 1-tert-butoxycarbonyl-3,6-dihydro-2H-pyridine-5-boronic acid pinacol ester were dissolved in 1,4-dioxane (10 mL) and water (2 mL). At room temperature, a Pd(dppf)Cl2 dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (135 mg, 1.27 mmol) were added. The mixture was purged with nitrogen three times, heated to 100 °C, and reacted for 1 hour. The reaction was confirmed to be complete by TLC. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to give a yellow solid, title compound 15a (260 mg, 89% yield).

[0685] LC-MS: m / z = 575.3 [M+H] +

[0686] Step 2: (R)-5-(4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-tert-butyl carboxylate 15b

[0687] Compound 15a (260 mg, 0.45 mmol) was dissolved in ethanol (10 mL) and water (3 mL). Reduced iron powder (126 mg, 2.26 mmol) and ammonium chloride (121 mg, 2.26 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 4 hours. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth filter, and the filtrate was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 15b (253 mg, crude product), which was used directly in the next step.

[0688] LC-MS: m / z = 545.3 [M+H] +

[0689] Step 3: (R)-5-(4-((1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-tert-butyl carboxylate 15c

[0690] Compound 15b (253 mg, crude) was dissolved in tetrahydrofuran (5 mL), and acetic anhydride (190 mg, 1.86 mmol) and N,N-diisopropylethylamine (360 mg, 2.79 mmol) were added at room temperature. The mixture was heated to 40 °C and reacted overnight. The reaction was confirmed to be complete by TLC. The reaction solution was cooled to room temperature, diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a brown solid, title compound 15c (250 mg, crude), which was used directly in the next step.

[0691] Step 4: 3-(4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 15d

[0692] Compound 15c (250 mg, crude) was dissolved in methanol (10 mL), and palladium / carbon (80 mg, 10%) was added at room temperature. The mixture was heated to 30 °C for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth liner, the filter cake was washed with methanol, and the filtrate was concentrated to give a yellow solid, title compound 15d (240 mg, crude), which was used directly in the next step.

[0693] Step 5: N-(3-((1R)-1-((8-methyl-7-oxo-6-(piperidin-3-yl)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 15e

[0694] Compound 15d (240 mg, crude) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 2 hours, and TLC showed that the starting material had completely reacted. The reaction solution was adjusted to alkalinity with saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 15e (180 mg, crude), which was used directly in the next step.

[0695] LC-MS: m / z = 489.3 [M+H] +

[0696] Step 6: N-(3-((1R)-1-((8-methyl-6-(1-methylpiperidin-3-yl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 15f

[0697] Compound 15e (180 mg, crude) was dissolved in methanol (5 mL), and an aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium / carbon (70 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 15f (142 mg, 63% yield in five steps).

[0698] Step 7: 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 15

[0699] Compound 15f (142 mg, 0.28 mmol) was dissolved in methanol (6 mL), and sodium hydroxide aqueous solution (3 mL, 12 mmol, 4 N) was added at room temperature. The mixture was heated to 70 °C and reacted overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 15 (80 mg, 62% yield).

[0700] LC-MS: m / z = 461.3 [M+H] +

[0701] 1H NMR (400MHz, CD3OD) δ8.33 (s, 1H), 8.22 (s, 1H), 6.96 (s, 1H), 6.93 (s, 1H), 6.80 (s, 1H), 5.50 (q, J=7.2Hz, 1H), 3.69 (s, 3H), 3.44-3.37 (m, 1H), 3.30-3.24(m, 2H), 2.70-2.57(m, 5H), 2.06-1.97(m, 2H), 1.91-1.81(m, 1H), 1.78-1.67(m, 1H), 1.62(d, J=7.2Hz, 3H). (99.76% purity by HPLC)

[0702] Example 16

[0703] 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 16

[0704]

[0705] Step 1 (R)-6-bromo-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one 16a

[0706] Compound 10f (150 mg, 0.55 mmol) was dissolved in dimethyl sulfoxide (3 mL). N,N-diisopropylethylamine (212 mg, 1.64 mmol) and intermediate IN-5 (80 mg, 0.42 mmol) were added at room temperature. The mixture was heated to 80 °C and reacted for 1 hour. TLC showed complete reaction of the starting material. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 16a (140 mg, 75% yield).

[0707] Step 2: (R)-5-(4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-tert-butyl carboxylate 16b

[0708] Compound 16a (140 mg, 0.33 mmol) and 6 g (141 mg, 0.46 mmol) of 1-tert-butoxycarbonyl-3,6-dihydro-2H-pyridine-5-boronic acid pinacol ester were dissolved in 1,4-dioxane (10 mL) and water (3 mL). Pd(dppf)Cl2 dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (97 mg, 0.92 mmol) were added at room temperature. The mixture was heated to 100 °C and stirred for 1 hour under nitrogen protection. The reaction was monitored by TLC until complete. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the brown oil title compound 16b (200 mg, crude product), which was used directly in the next step.

[0709] LC-MS: m / z = 530.3 [M+H] +

[0710] Step 3: tert-butyl 3-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylate 16c

[0711] Compound 16b (200 mg, crude) was dissolved in methanol (10 mL), and palladium / carbon (50 mg, 10%) was added at room temperature. The mixture was heated to 30 °C for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was cooled to room temperature, filtered through a diatomaceous earth liner, the filter cake was washed with methanol, and the filtrate was concentrated to give a yellow solid, title compound 16c (220 mg, crude), which was used directly in the next step.

[0712] LC-MS: m / z = 532.3 [M+H] +

[0713] Step 4: 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-(piperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 16d

[0714] Compound 16c (220 mg, crude) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 2 hours, and TLC showed that the starting material had completely reacted. The reaction solution was adjusted to alkalinity with saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 16d (210 mg, crude), which was used directly in the next step.

[0715] Step 5: 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 16

[0716] Compound 16d (210 mg, crude) was dissolved in methanol (5 mL), and an aqueous formaldehyde solution (125 mg, 1.54 mmol, 37%) and palladium / carbon (70 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the starting material had completely reacted. The reaction solution was filtered, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 16 (70 mg, 48% yield in four steps).

[0717] LC-MS: m / z = 446.3 [M+H] +

[0718] 1 H NMR (400MHz, CD3OD) δ8.28 (s, 1H), 8.21 (s, 1H), 7.57 (t, J=7.2Hz, 1H), 7.47 (t, J=6.8Hz, 1H), 7.23 (t, J=7.6Hz, 1H), 7.14-6.86 (m, 1H), 5.78 (t, J=7.2Hz, 1H), 3 .68(s, 3H), 3.30-3.19(m, 2H), 3.14-3.08(m, 1H), 2.48(s, 3H), 2.37-2.25(m, 2 H), 2.04-1.76 (m, 3H), 1.66 (d, J=6.8Hz, 3H), 1.64-1.56 (m, 1H). (97.17% purity by HPLC)

[0719] Example 17

[0720] 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((S)-1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17-1

[0721] 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((R)-1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17-2

[0722]

[0723] Step 1: 3-(2-ethoxy-2-oxoethyl)piperidine-1-carboxylic acid tert-butyl ester 17b

[0724] N-Boc-3-piperidineacetic acid 17a (5.0 g, 20.55 mmol) was dissolved in N,N-dimethylformamide (50 mL), and potassium carbonate (8.5 g, 61.65 mmol) and ethyl iodine (4.8 g, 30.82 mmol) were added. The reaction mixture was reacted at room temperature for 5 hours. The reaction solution was extracted with water and ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a yellow oily title compound 17b (5.2 g, 93% yield).

[0725] Step 2: 3-(4-chloro-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidin-1-carboxylic acid tert-butyl ester 17c

[0726] Compound 13a (1.13 g, 6.59 mmol) was dissolved in tetrahydrofuran (70 mL), and compound 17b (2.32 g, 8.56 mmol) was added. The mixture was cooled to -15 °C, and lithium diisopropylamino (6.6 mL, 13.17 mmol, 2.0 M) was added dropwise. After the addition was complete, the mixture was reacted at -10 °C for 5 hours, then allowed to rise to room temperature and reacted overnight. TLC showed that the starting material had completely reacted. The reaction solution was cooled to 0 °C, quenched with saturated citric acid aqueous solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to silica gel column chromatography to obtain a crude product. The crude product was slurried with petroleum ether, filtered, and the yellow solid title compound 17c (840 mg, yield 34%) was obtained.

[0727] LCMS: m / z = 323.1 [M+H-56] +

[0728] Step 3: (S)-3-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 17d-1 & (R)-3-(4-((((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 17d-2

[0729] Compound 17c (421 mg, 1.11 mmol) was dissolved in dimethyl sulfoxide (4 mL). Intermediate IN-5 (200 mg, 0.10 mmol) and N,N-diisopropylethylamine (430 mg, 3.33 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted for 3 hours. TLC showed the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with 1N hydrochloric acid, then with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 17d (310 mg, 52% yield). Compound 17d was chirally resolved (Daicel AD-H, 30*250mm, 5μm, 30mL / min, IPA:Hexane = 5:95) to yield a pale yellow solid, 17d-1 (peak 1, RT 7.95min) (100mg, yield 17%) and a pale yellow solid, 17d-2 (peak 2, RT 11.33min) (114mg, yield 20%). Further analysis is needed to determine the configuration and properties of the compounds; 17d-1 and 17d-2 are tentatively identified as having the above configurations.

[0730] LCMS: m / z = 532.2 [M+H]

[0731] Step 4: 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((R)-piperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17e-1 & 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((R)-piperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17e-2

[0732] Compound 17d-1 (100 mg, 0.19 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction was carried out at room temperature for 2 hours, and TLC showed that the reaction was complete. The reaction solution was adjusted to alkalinity by adding saturated sodium bicarbonate aqueous solution dropwise, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 17e-1 (100 mg, crude product), which was used directly in the next step.

[0733] Compound 17d-2 (114 mg, 0.21 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction was carried out at room temperature for 2 hours, and TLC showed that the reaction was complete. The reaction solution was adjusted to alkalinity by adding saturated sodium bicarbonate aqueous solution dropwise, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 17e-2 (85 mg, crude product), which was used directly in the next step.

[0734] The configuration and properties of the compounds need further investigation; 17e-1 and 17e-2 are tentatively identified as the above configurations.

[0735] Step 5: 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((S)-1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17-1 & 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((R)-1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17-2

[0736] Compound 17e-1 (100 mg, crude) was dissolved in ethanol (5 mL), and an aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium / carbon (40 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 17-1 (55 mg, 65% yield in two steps).

[0737] Compound 17e-2 (85 mg, crude) was dissolved in ethanol (5 mL), and an aqueous formaldehyde solution (85 mg, 1.05 mmol, 37%) and palladium / carbon (40 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 17-2 (47 mg, 50% yield in two steps).

[0738] The configuration and properties of the compounds need further investigation; 17-1 and 17-2 are tentatively identified as the above configurations.

[0739] 17-1:

[0740] LCMS: m / z = 446.2[M+H]+

[0741] 1H NMR (400MHz, CD3OD) δ8.29 (s, 1H), 8.20 (s, 1H), 7.57 (t, J=7.2Hz, 1H), 7.47 ( t, J=7.2Hz, 1H), 7.23 (t, J=7.6Hz, 1H), 6.99 (t, J=54.8Hz, 1H), 5.78 (q, J=6. 8Hz, 1H), 3.68 (s, 3H), 3.30-3.19 (m, 2H), 3.14-3.07 (m, 1H), 2.47 (s, 3H), 2. 36-2.25(m, 2H), 2.03-1.77(m, 3H), 1.66(d, J=7.2Hz, 3H), 1.65-1.56(m, 1H).

[0742] 17-2:

[0743] LCMS: m / z = 446.2 [M+H] +

[0744] 1 H NMR (400MHz, CD3OD) δ8.29 (s, 1H), 8.19 (s, 1H), 7.57 (t, J=7.6Hz, 1H), 7.47 (t, J= 7.2Hz, 1H), 7.23 (t, J=7.6Hz, 1H), 6.99 (t, J=54.8Hz, 1H), 5.78 (q, J=6.8Hz, 1H), 3.69(s, 3H), 3.30-3.16(m, 2H), 3.11-3.04(m, 1H), 2.45(s, 3H), 2.33-2.20(m, 2H ), 2.03-1.96(m, 1H), 1.95-1.76(m, 2H), 1.66(d, J=7.2Hz, 3H), 1.63-1.55(m, 1H).

[0745] Example 18

[0746] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-((R)(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 18-1

[0747] 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-((S)(1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 18-2

[0748]

[0749] The compounds in Example 6 were chirally resolved (Daicel AD-H, 30*250mm, 5μm, 30mL / min, IPA:Hexane = 5:95) to yield solid title compounds 18-1 and 18-2. The configurations and properties of these compounds require further investigation; 18-1 and 18-2 are tentatively identified as having the above configurations.

[0750] 18-1:

[0751] LC-MS: m / z = 475.3 [M+H] +

[0752] 1 H NMR (400MHz, Methanol-d4) δ8.13 (s, 1H), 6.96 (s, 2H), 6.80 (s, 1H), 5.55 (q, J=7.0Hz, 1H), 3.68 (s, 3H), 3.27 (d, J=21.5Hz, 2H ), 3.20 (d, J=11.5Hz, 1H), 2.57 (s, 3H), 2.45 (d, J=13.9Hz, 5H), 2.03-1.94 (m, 2H), 1.85 (d, J=13.1Hz, 1H), 1.72-1.56 (m, 4H).

[0753] 18-2:

[0754] LC-MS: m / z = 475.3 [M+H] +

[0755] 1 H NMR (400MHz, Methanol-d4) δ8.10 (s, 1H), 6.95 (d, J=5.5Hz, 2H), 6.80 (s, 1H), 5.55 (d, J=7.1Hz, 1H), 3.69 (s, 3H), 3.29-3.19 (m, 2H), 3.11 (d, J =11.9Hz, 1H), 2.48 (s, 3H), 2.44 (s, 3H), 2.32 (q, J = 11.6Hz, 2H), 1.94 (dd, J = 19.9, 13.4Hz, 2H), 1.83 (t, J = 12.8Hz, 1H), 1.60 (d, J = 7.1Hz, 4H).

[0756] Example 19

[0757] 8-Methyl-6-(1-methylpiperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 19

[0758]

[0759] The compound of Example 19 was synthesized according to the synthesis method of Example 20.

[0760] Example 20

[0761] 8-Methyl-6-((S)-1-methylpiperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 20-1

[0762] 8-Methyl-6-((R)-1-methylpiperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 20-2

[0763]

[0764] Step 1 (S)-3-(8-methyl-7-oxo-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 20a-1 & (R)-3-(8-methyl-7-oxo-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl ester 20a-2

[0765] Compound 17c (341 mg, 0.90 mmol) was dissolved in dimethyl sulfoxide (4 mL). Intermediate IN-8 (153 mg, 0.81 mmol) and N,N-diisopropylethylamine (348 mg, 2.70 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted for 3 hours. TLC showed the reaction was complete. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, washed with dilute hydrochloric acid (1 N) and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 20a (198 mg, 46% yield). Compound 20a was chirally resolved (Daicel AD-H, 30*250mm, 5μm, 30mL / min, IPA:Hexane = 5:95) to give pale yellow solid compound 20a-1 (peak 1, RT 8.75min) (85mg, yield 20%) and pale yellow solid compound 20a-2 (peak 2, RT 8.53min) (94mg, yield 22%). The configuration and properties of the compounds require further investigation; 20a-1 and 20a-2 are tentatively identified as having the above configurations.

[0766] LCMS: m / z = 532.2 [M+H] +

[0767] Step 2: 8-Methyl-6-((S)-piperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 20b-1 & 8-Methyl-6-((R)-piperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 20b-2

[0768] Compound 20a-1 (85 mg, 0.16 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction mixture was reacted at room temperature for 2 hours, and the reaction was observed until the reaction was complete by TLC. The reaction mixture was adjusted to alkalinity by adding saturated sodium bicarbonate solution dropwise, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 20b-1 (100 mg, crude product), which was used directly in the next step.

[0769] Compound 20a-2 (94 mg, 0.17 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (2 mL) was added. The reaction was carried out at room temperature for 2 hours, and TLC showed that the reaction was complete. The reaction solution was adjusted to alkalinity by adding saturated sodium bicarbonate aqueous solution dropwise, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow solid, title compound 21b-2 (105 mg, crude product), which was used directly in the next step.

[0770] The configuration and properties of the compounds need further investigation; 20b-1 and 20b-2 are tentatively identified as having the above configurations.

[0771] Step 3: 8-Methyl-6-((S)-1-methylpiperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 20-1 & 8-Methyl-6-((R)-1-methylpiperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 20-2

[0772] Compound 20b-1 (100 mg, crude) was dissolved in ethanol (5 mL), and an aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium / carbon (40 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, and the filtrate was concentrated to give pale yellow solids, title compounds 20-1 and 20-2 (55 mg, 65% yield in two steps).

[0773] Compound 21b-2 (105 mg, crude) was dissolved in ethanol (5 mL), and an aqueous formaldehyde solution (105 mg, 1.29 mmol, 37%) and palladium / carbon (45 mg, 10%) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere, and TLC showed that the reaction was complete. The reaction solution was filtered through a diatomaceous earth filter, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a pale yellow solid, title compound 20-2 (50 mg, two-step yield 63%).

[0774] The configuration and properties of the compounds need further investigation; 20-1 and 20-2 are tentatively identified as the above configurations.

[0775] 20-1:

[0776] LCMS: m / z = 446.2[M+H]+

[0777] 1 H NMR (400MHz, CD3OD) δ8.32 (s, 1H), 8.19 (s, 1H), 7.73-7.66 (m, 2H), 7.55-7.47 (m, 2H), 5.60 (q, J=6.8Hz, 1H), 3.70 (s, 1H), 3.37-3.26 (m, 1H), 3.26-3.19(m, 1H), 2.59(s, 3H), 2.56-2.44(m, 2H), 2.05-1.94(m, 2H), 1.93-1.79(m, 1H), 1.76-1.67(m, 1H), 1.67(d, J=6.8Hz, 3H).

[0778] 20-2:

[0779] LCMS: m / z = 446.2 [M+H] +

[0780] 1 H NMR (400MHz, CD3OD) δ8.32 (s, 1H), 8.19 (s, 1H), 7.76-7.67 (m, 2H), 7.56-7.45 (m, 2H), 5.60 (q, J=6.8Hz, 1H), 3.69 (s, 3H), 3.38-3.26 (m, 1H), 3.26-3.19(m, 1H), 2.59(s, 3H), 2.58-2.43(m, 2H), 2.06-1.94(m, 2H), 1.92-1.80(m, 1H), 1.75-1.68(m, 1H), 1.67(d, J=7.2Hz, 3H).

[0781] Example 21

[0782] 4-(((R)-1-(5-amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((1-methylpyrrolidone-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 21

[0783]

[0784] Referring to Example 14, Example 21 was synthesized.

[0785] Example 22

[0786] 2,8-Dimethyl-4-(((R)-1-(4-(2-(((methylamino)methyl)phenyl)thiophene-2-yl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 22

[0787]

[0788] Step 1 (2-(5-((R)-1-((2,8-dimethyl-7-oxo-6-(((S)-tetrahydrofuran-3-yl)oxy)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)thiophene-3-yl)benzyl)(methyl)benzyl carbamate 22a

[0789] Compound 1f (129 mg, 0.44 mmol) was dissolved in N-methylpyrrolidone (3 mL). Intermediate IN-2 (119 mg, 0.31 mmol) and triethylamine (126 mg, 1.24 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted for 4 hours. TLC showed that the starting material had completely reacted. The reaction solution was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 22a (30 mg, 15% yield).

[0790] Step 2: 2,8-Dimethyl-4-(((R)-1-(4-(2-(((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 22

[0791] Compound 22a (30 mg, 0.047 mmol) was dissolved in methanol (5 mL), and palladium / carbon (30 mg, 10%) was added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a white solid title compound 22 (14 mg, 60% yield).

[0792] LC-MS: m / z = 506.2 [M+H]+

[0793] 1 H NMR (400MHz, CD3OD) δ7.51 (s, 1H), 7.48-7.42 (m, 1H), 7.39-7.28 (m, 3H), 7.21 (d, J=1.2Hz, 1H), 7.12 (s, 1H), 5.99 (q, J=6.8Hz, 1H), 5.09-5.06 (m, 1H), 4.03-3.94 (m, 3H), 3.92-3.84 (m, 3H), 3.73 (s, 3H), 2.50 (s, 3H), 2.35 (s, 3H), 2.32-2.14 (m, 2H), 1.77 (d, J=7.2Hz, 3H). (89.04% purity by HPLC)

[0794] Example 23

[0795] 2-Methyl-4-(((R)-1-(4-(2-(((methylamino)methyl)phenyl)thiophene-2-yl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 23

[0796]

[0797] Step 1: methyl(2-(5-((R)-1-((2-methyl-7-oxo-6-(((S)-tetrahydrofuran-3-yl)oxy)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)thiophene-3-yl)phenyl)benzyl carbamate 23a

[0798] Compound 2e (221 mg, 0.78 mmol) was dissolved in dimethyl sulfoxide (3 mL). Intermediate IN-2 (300 mg, 0.79 mmol) and N,N-diisopropylethylamine (356 mg, 2.75 mmol) were added at room temperature. The mixture was heated to 100 °C and reacted overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give a yellow solid, title compound 23a (140 mg, 29% yield).

[0799] LC-MS: m / z = 626.3 [M+H] +

[0800] Step 2: 2-Methyl-4-(((R)-1-(4-(2-(((methylamino)methyl)phenyl)thiophene-2-yl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 23

[0801] Compound 23a (140 mg, 0.22 mmol) was dissolved in methanol (5 mL), and palladium / carbon (30 mg, 10%) and concentrated hydrochloric acid (2 drops) were added. The reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give a white solid, title compound 23 (53 mg, yield 49%).

[0802] LC-MS: m / z = 492.2 [M+H] +

[0803] 1 H NMR (400MHz, CD3OD) δ7.53 (s, 1H), 7.49-7.44 (m, 1H), 7.39-7.29 (m, 3H), 7.21 (s, 1H), 7.13 (s, 1H), 5.98 (q, J=6.8Hz, 1 H), 5.07 (s, 1H), 4.05-3.83 (m, 6H), 2.46 (s, 3H), 2.37 (s, 3H), 2.32-2.13 (m, 2H), 1.77 (d, J=6.8Hz, 3H). (91.42% purity by HPLC)

[0804] Example 24

[0805] 8-Methyl-4-(((R)-1-(4-(2-(((methylamino)methyl)phenyl)thiophene-2-yl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 24

[0806]

[0807] Step 1 (2-((methylamino)methyl)phenyl)boronic acid 24a

[0808] o-Formylboronic acid IN-2j (5.0 g, 33.35 mmol) was dissolved in methylamine ethanol solution (30 mL, 7 M), and palladium on carbon (200 mg, 10%) was added. The reaction was carried out at room temperature for 5 hours under a hydrogen atmosphere, and the reaction was confirmed to be complete by TLC. The reaction solution was filtered through diatomaceous earth, the filter cake was washed, and the filtrate was concentrated to obtain a pale yellow bubbly solid, title compound 24a (5.3 g, crude product), which was used directly in the next step.

[0809] Step 2 (2-(((tert-butoxycarbonyl)(methyl)-14-azayl)methyl)phenyl)boronic acid 24b)

[0810] Compound 24a (5.3 g, crude) was dissolved in tetrahydrofuran (100 mL) and water (20 mL). Sodium carbonate (10.3 g, 97.18 mmol) was added at room temperature, and the mixture was cooled to 0 °C. Di-tert-butyl dicarbonate (8.5 g, 38.95 mmol) was added dropwise. After the addition was complete, the mixture was allowed to return to room temperature and reacted overnight. The reaction was confirmed to be complete by TLC. The reaction solution was extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and then slurried with crude petroleum ether and ethyl acetate (5 / 1). The mixture was filtered, and the filter cake was washed and dried to give a white solid, title compound 24b (7.4 g, 84% yield in two steps).

[0811] 1 H NMR (400MHz, DMSO-d6) δ8.14 (s, 2H), 7.50 (dd, J=7.2, 0.8Hz, 1H), 7.38-7.30 (m, 1H), 7.21 (t, J=7.2Hz, 1H), 7.08 (d, J=7.6Hz, 1H), 4.56 (s, 2H), 2.74 (s, 3H), 1.40 (s, 9H).

[0812] Step 3: (R)-(2-(5-(1-aminoethyl)thiophen-3-yl)benzyl)(methyl)carbamate tert-butyl ester 24c

[0813] Compound IN-2e (1.0 g, 4.85 mmol) and compound 24b (1.9 g, 7.17 mmol) were dissolved in 1,4-dioxane (20 mL) and water (5 mL). Sodium carbonate (1.5 g, 14.15 mmol) and a Pd(dppf)Cl2 dichloromethane complex (200 mg, 0.24 mmol) were added at room temperature. The mixture was purged with nitrogen three times and heated to 100 °C for 4 hours. The reaction was confirmed to be complete by TLC. The reaction solution was cooled to room temperature, extracted with water and ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography to give the red oily compound 24c (800 mg, 48% yield).

[0814] LC-MS: m / z = 347.2 [M+H] +

[0815] Step 4: (S)-4-chloro-8-methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 24d

[0816] Compound 13a (12.0 g, 6.99 mmol) was dissolved in tetrahydrofuran (180 mL). Under nitrogen protection, intermediate IN-1 (18.3 g, 0.11 mol) was added, and the mixture was cooled to -60 °C. Lithium bis(trimethylsilylamino)amine (17.5 mL, 0.175 mol, 1.0 M tetrahydrofuran solution) was added dropwise. After the addition was complete, the mixture was reacted at -10 °C for 5 hours. TLC showed that the intermediate had completely reacted, with a small amount of the starting material remaining unreacted. The reaction mixture was quenched dropwise with saturated citric acid solution, adjusted to acidity, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 24d (7.2 g, 36% yield) as a brownish-yellow solid.

[0817] LCMS: m / z = 282.1 [M+H] +

[0818] Step 5: tert-butylmethyl(2-(5((R)-1-((8-methyl-7-oxo-6-(((S)-tetrahydrofuran-3-yl)oxy)-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)thiophene-3-yl)benzyl)carbamate 24e

[0819] Compound 24d (81 mg, 0.29 mmol) was dissolved in dimethyl sulfoxide (3 mL). Compound 24c (100 mg, 0.29 mmol) and N,N-diisopropylethylamine (111 mg, 0.86 mmol) were added at room temperature. The mixture was heated to 90 °C and reacted overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 24e (80 mg, 47% yield).

[0820] LCMS: m / z = 592.3 [M+H] +

[0821] Step 6: 8-Methyl-4-(((R)-1-(4-(2-(((methylamino)methyl)phenyl)thiophene-2-yl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 24

[0822] Compound 24e (80 mg, 0.14 mmol) was dissolved in methanol hydrochloric acid (3 mL, 4 N) and reacted at room temperature for 2 hours. TLC showed that the reaction was complete. The pH of the reaction mixture was adjusted to alkaline by adding saturated sodium bicarbonate solution dropwise, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to give a yellow solid, title compound 24 (45 mg, 67% yield).

[0823] LCMS: m / z = 492.2 [M+H] +

[0824] 1 H NMR (400MHz, CD3OD) δ8.37 (s, 1H), 7.53 (s, 1H), 7.40-7.47 (m, 1H), 7.39-7.2 6(m, 3H), 7.21(s, 1H), 7.13(s, 1H), 5.94(t, J=6.4Hz, 1H), 5.15-5.06(m, 1H), 4.07-3.94(m, 3H), 3.93-3.85(m, 1H), 3.83(s, 2H), 3.74(s, 3H), 2.32(s, 3H) , 2.30-2.24(m, 1H), 2.24-2.15(m, 1H), 1.78(d, J=6.8Hz, 3H). (97.81% purity (by HPLC)

[0825] Test Example 1: Inhibitory activity of compound against SOS1

[0826] Experimental steps:

[0827] 1. Compound preparation: Prepare a compound to a final concentration of 400 times. For example, if the final concentration to be measured is 5 μM, prepare a compound to a final concentration of 400 times, i.e., 2 mM. Use an automated micropipette to serially dilute the compound to the set number of concentration points.

[0828] 2. Transferring the compound to a 384-well plate: Using an ultrasonic nanoliter liquid handling system, transfer 50 nL of the diluted compound from the Echo 384-well plate to a 384-well plate. For both the negative and positive controls, transfer 50 nL of 100% DMSO.

[0829] 3. Prepare and transfer 4-fold Tag1-SOS1 solution: Prepare 4-fold Tag1-SOS1 solution using Diluent provided in the kit (KRAS-G12C / SOS1 BINGDINGASSAYKIT (Cisbio, Cat.No.63ADK000CB16PEG)), transfer 5 μL to a 384-well plate, and transfer 5 μL of Diluent replacement enzyme solution to the negative control wells. Centrifuge at 1000 rpm for 1 minute.

[0830] 4. Prepare 4-fold Tag2-KRAS G12C solution: Prepare 4-fold Tag2-KRAS G12C solution using the Diluent provided in the kit, transfer 5 μL to a 384-well reaction plate, and centrifuge at 1000 rpm for 1 minute.

[0831] 5. Transfer 2x detection solution: Prepare 2x Anti-Tag1-Tb3+ and Anti-Tag2-XL665 solutions using the Detection Buffer provided in the kit, transfer 10 μL to a 384-well reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 60 minutes.

[0832] 6. Reading: Use an Envision microplate reader to read the fluorescence signal values ​​(Ex665 / Em615).

[0833] 7. Inhibition rate calculation and IC50 fitting

[0834] The values ​​are copied from the plate reader, where the maximum value refers to the reading of the positive control and the minimum value refers to the reading of the negative control. Inhibition rate (%) = (maximum value - sample value) / (maximum value - minimum value) × 100%.

[0835] Import the data into MS Excel and fit the IC using XLFit Excel add-in version 5.4.0.8. 50 Values; results are shown in Table 1.

[0836] Fitting formula: Y = Bottom + (Top - Bottom) / (1 + (IC50 / X)^HillSlope)

[0837] Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> 1 8.5 2 9.7 3 8.0 4 6.5 5 6.1 6 5.9 7 5.3 8 12.0 9 13.0 10 5.0 13 4.6 14 5.7 17-1 5.1 17-2 9.3 18-1 4.9 18-2 11 20-1 8.3 20-2 15 22 8.0 23 5.0 24 5.8

[0838] IC50 of compound 2 on the inhibition of K-562 cell proliferation 50 Measurement

[0839] The human chronic myeloid leukemia cells K-562 (CCL-243) used in this invention were purchased from the American Type Culture Collection (ATCC). The cells were grown in RPMI 1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin antibiotics at 37°C and 5% CO2.

[0840] The inhibitory effect of the compound on the proliferation of K-562 cells in vitro was determined by the following methods:

[0841] 1) Cell seeding: 20,000 K-562 cells in good logarithmic growth phase were seeded into 96-well plates at 90 μL per well and cultured at 37°C and 5% CO2 for 24 hours.

[0842] 2) Drug addition: The compound to be tested was serially diluted with complete culture medium. 10 μL of the diluted compound was added to 90 μL of cells to achieve final concentrations of 10000, 3000, 1000, 300, 100, 30, 10, 3, and 1 nM. Corresponding solvent controls were also provided. Cells were incubated at 37°C in a 5% CO2 incubator for 96 hours.

[0843] 3) Detection: Add 10 μL of 5 mg / mL MTT working solution (ABCONE, M9609) to each well, incubate at 37°C for 4 hours, then add triple solution (10% SDS, 0.5% isopropanol, 0.1 mol / L HCl) until the cell lysate is completely dissolved, and read the OD570 and OD690 values ​​using a TECAN SPARK microplate reader.

[0844] 4) Calculation: Calculate the cell growth inhibition rate using the following formula:

[0845] Inhibition rate = (control wells) OD570nm-OD690nm - Drug delivery port OD570nm-OD690nm ) / Reference Hole OD570nm-OD690nm ×100%

[0846] IC50 was calculated using Graphpad Prism 5.0 software based on compound concentration and corresponding inhibition rate. 50 Values. The experimental results are shown in Table 2.

[0847] Compound numbering K-562 Compound numbering K-562 BI-3406 35.2 13 99.9 1 461.7 15 17.0 2 246.9 16 38.3 3 118.3 17-1 28 4 99.9 17-2 148.8 5 75.6 18-1 50.7 6 94.9 18-2 206.8 8 288.8 20-1 29.5 9 326.5 20-2 198.3 10 34.0 22 333 11 37.1 23 274.3

[0848] Conclusion: The compounds in the embodiments of the present invention have an inhibitory effect on the proliferation of K-562 cells, and the activities of several compounds are comparable to those of BI-3402.

[0849] Effect of Compound 3 on Phosphorylation Level of ERK1 / 2, a Downstream Signaling Molecule of KRAS, in K-562 Cells

[0850] The effect of the compounds of this invention on ERK1 / 2 phosphorylation levels in K-562 cells was determined by the following methods:

[0851] 1) Cell seeding: Take K-562 cells in good logarithmic growth phase and seed them at a rate of 1*10-1. 6 Inoculate one well per cell into a six-well plate and incubate overnight at 37°C and 5% CO2.

[0852] 2) Drug addition: The compound to be tested was serially diluted with complete culture medium and added to the cells to achieve final concentrations of 1000, 100, 10, and 1 nM. The cells were then incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours.

[0853] 3) Protein sample preparation: Collect cell suspension, centrifuge at 500g for 5 minutes, discard supernatant, wash 3 times with PBS, and lyse cells with 100 μL of 1×SDS gel loading buffer (50 mM Tris-HCl (pH 6.8), 100 mM DTT, 2% SDS, 10% glycerol, 0.1% bromophenol blue). Denature the cell lysate by heating at 100℃ for 10 minutes.

[0854] 4) Western blot: Protein samples were subjected to SDS-PAGE electrophoresis. After electrophoresis, the proteins were transferred to a PVDF membrane using a wet transfer system. The PVDF membrane was then blocked in blocking buffer (5% skim milk powder diluted in TBS / T) at room temperature for 1 hour, followed by reaction with antibodies I and II. After washing the membrane, staining was performed using Immobilon Western HRP Substrate Luminal Reagent, and images were taken using a Western Blot imaging system (Tanon, 4600). The following is the antibody information used: p-ERK1 / 2 (CST: 4370); ERK1 / 2 (CST: 9102); β-tubulin (CST: 2146).

[0855] The results of the effect of the compound on ERK1 / 2 phosphorylation levels in K-562 cells are shown in the figure. Figure 1 .

[0856] Conclusion: In the embodiments of the present invention, compound 5 has a significant inhibitory effect on the phosphorylation of ERK1 / 2 in K-562 cells. The inhibitory activity is concentration gradient dependent and is roughly equivalent to that of BI-3402.

[0857] Test Example 4: Stability of Compounds to Liver Microsomes in Mice and Humans

[0858] Experimental steps:

[0859] (1). Take out liver microsomes (20mg protein / mL) from the -80℃ freezer, place them on a 37℃ water bath constant temperature shaker for 3 minutes to thaw and set aside for use.

[0860] (2) Prepare a mixed solution of the incubation system (excluding β-NADPH) according to the proportions in the "Composition of Experimental Incubation System" above.

[0861] (3) Prepare a 100 μM working solution of the test compound for later use.

[0862] (4). Control group (without β-NADPH): Take 25 μL of PB solution into 75 μL of the incubation system mixture described in (2), vortex for 30 s, mix well, and the total reaction volume is 100 μL. Repeat the sample. Place it in a 37℃ water bath constant temperature shaker for incubation and start timing. The sampling time points are 0 min and 60 min.

[0863] (5). Sample group: Take 25 μL of β-NADPH solution (4 mM) and add it to 75 μL of the reaction system described in (2). Vortex for 30 s to mix well. The total reaction volume is 100 μL. Repeat the sample. Place it in a 37℃ water bath constant temperature shaker for incubation and start timing. The sampling time points are 0 min, 5 min, 15 min, 30 min, and 60 min.

[0864] (6) At each time point, remove the sample tube and add 300 μL of cold stop agent (including internal standard) to terminate the reaction.

[0865] (7) Vortex and centrifugal.

[0866] (8) Take 150 μL of the supernatant and add 150 μL of water, vortex to mix, and then analyze by LC-MS / MS.

[0867] Data Analysis: Calculate the half-life (t) using the following first-order kinetic formula. 1 / 2 ) and clearance rate (CL)

[0868] C t =C0*e -kt

[0869] C t = (1 / 2)*C0

[0870] t 1 / 2 =ln2 / k=0.693 / k

[0871] CL=V d *k

[0872] Vd = 1 / Protein content in liver microsomes

[0873] CL int(liver) =CL int(mic) × Liver weight to body mass ratio × Liver microsomal protein concentration per gram of liver

[0874] The parameters in the formula are shown in Table 3:

[0875] Table 3 Common parameters of liver and blood in mice, rats, and humans.

[0876]

[0877] The experimental results are shown in Table 4:

[0878] Table 4. Liver microsomal stability of compounds in different species.

[0879]

[0880] In human liver microsomal stability experiments, compounds 2, 3, 4, and 13 showed significantly better stability than BI-3406, while compound 5 showed stability comparable to BI-3406. In rat liver microsomal stability experiments, compounds 3, 5, 6, and 13 showed significantly better stability than BI-3406, while compound 4 showed stability comparable to BI-3406. In mouse liver microsomal stability experiments, compounds 2, 3, 4, 5, 6, 13, and 18 showed significantly better stability than BI-3406. In summary, the liver microsomal stability of several compounds in the examples was significantly better than that of BI-3406 in different species.

[0881] Study on CYP enzyme inhibition of compound in Test Example 5

[0882] Experimental steps:

[0883] (1) 100× specific inhibitor: take the corresponding stock solution and dilute it with 50% acetonitrile-water to prepare the corresponding concentration of inhibitor working solution;

[0884] (2) 100× compound: Dilute the compound stock solution with pure acetonitrile to prepare a 2000 μM working solution;

[0885] (3) 200× substrate: Take the corresponding stock solution and dilute it with 50% acetonitrile-water to prepare a substrate working solution of the corresponding concentration;

[0886] (4) Use PB solution as a solvent to prepare a 4 mM NADPH solution;

[0887] (5) Mix 4 in l mixed solution: Take a certain amount of PB into a centrifuge tube, add a certain amount of MgCl2-PB (6mM) solution, then add human liver microsomes (20mg / mL) solution, substrate working solution (Phe: 1A2, 18mM; Bup: 2B6, 12mM; Amo: 2C8, 200μM; Tes: 3A4, 4mM), vortex to mix, and dispense into 148μL / tube.

[0888] (6) Mix 3 in 1 solution: Take a certain amount of PB into a centrifuge tube, add a certain amount of MgCl2-PB (6mM) solution, then add human liver microsomes (20mg / mL) solution, substrate working solution (Dic: 2C9, 1.6mM; DM: 2D6, 800μM; Mid: 3A, 400μM), vortex to mix, and dispense into 148μL / tube.

[0889] (7) Liver microsome substrate mixture 2C19: Take a certain amount of PB into a centrifuge tube, add a certain amount of MgCl2-PB (6mM) solution, then add human liver microsome (20mg / mL) solution and substrate working solution (Mep: 2C19, 4mM), vortex to mix, and dispense into 148μL / tube.

[0890] (8) Add 1 μL of analyte working solution / 50% acetonitrile-water to each tube of the test sample, and then add 1 μL of test sample working solution / 50% acetonitrile-water to each tube of the inhibitor group. Then place them in a 37°C water bath for 5 min for pre-incubation. At the same time, pre-incubate NADPH in a 37°C water bath for 5 min.

[0891] (9) Add 50 μL / well of NADPH working solution and incubate for 30 min (S-Mefphenytoin) and 10 min (other substrates);

[0892] (10) Add 600 μL / well of ice internal standard working solution, vortex for 5 min to terminate the reaction, and centrifuge at 5500g for 10 min;

[0893] (11) Take 100 μL of the supernatant and add 300 μL of water (phenacetin and amodiaquine), take 150 μL of the supernatant and add 150 μL of water (other substrate groups), vortex to mix, and analyze by LC-MS / MS.

[0894] Data Analysis

[0895] The inhibition rates of the analyte and control at different concentration levels on the metabolic rates of each specific probe substrate were calculated using Excel. Then, the IC50 was calculated using the following formula. 50 The calculation results are shown in Table 5:

[0896] *IC 50 = x*(100-enzyme inhibition rate%at x) / enzyme inhibition rate%, assuming Hillslope=1.

[0897] x represents the inhibitor concentration.

[0898] Table 5. Inhibitory activity of the compounds in the examples against CYP enzymes.

[0899]

[0900] Generally speaking, for each subtype of CYP450, IC 50 >10 μmol / L can be considered as weak CYP inhibition; 3 μmol / L <IC50 50 <10 μmol / L can be considered as moderate inhibition of CYP; IC50 50A concentration <3 μmol / L is considered a strong inhibitor of CYP. BI-3406 exhibited moderate inhibitory activity against multiple CYP450 isoforms, including 2C9, 2C19, 2D6, and 3A. However, compounds 2, 3, 13, 18, 23, and 24 showed weak inhibition against the 2C9 isoform; compounds 2, 3, 5, 6, 13, 18, 23, and 24 showed weak inhibition against the 2C19 isoform; compounds 2, 3, 4, 5, 6, 8, 13, and 18 showed weak inhibition against the 2D6 isoform; and compounds 3, 6, 13, 18, 23, and 24 showed weak inhibition against the 3A isoform. The tested compounds all showed weak inhibitory activity against 1A2, 2B6, and 2C8. In summary, several compounds showed weaker inhibition of CYP450 than BI-3406, suggesting a potentially lower risk of drug interactions and better drug safety.

[0901] Test Example 6: In vivo antitumor activity of compound

[0902] Female BALB / c mice (6-8 weeks old) were subcutaneously injected with MIAPaCa-2 or HPAF-II cells (1.0 × 10⁻⁶ cells) on the right side. 7 (Cells / Mouse). Mice were administered compound 5 (50 mg / kg, bid), MRTX849 (5 mg / kg, qd), Trametinib (0.125 mg / kg, bid), and combinations of compound 5 with Trametinib or MRTX849 via gavage. Mice were monitored daily and caliper measurements were initiated when tumors became visible. Tumor volume was calculated by measuring two vertical diameters using the following formula: (L*W²) / 2, where L and W refer to the length and width of the tumor diameter. When the average tumor volume reached 100 mm², the tumor was considered complete. 3 Mice were divided into groups (D0, n = 5 / group) and administered the compound. Tumor volume and mouse body weight were measured every 3 days during the administration period. Results are shown in […]. Figure 2-5 .

[0903] Conclusions: Compound 5 of the present invention, when combined with the KRAS G12C inhibitor MRTX849, significantly enhanced the tumor inhibition rate against MIAPaCa-2 (KRASG12C) xenografts in nude mice. The tumor inhibition rates of Compound 5 (50 mg / kg, bid) monotherapy, MRTX849 (5 mg / kg, qd) monotherapy, and the combination therapy were 71.2%, 60.8%, and 98.1%, respectively (P < 0.001). Compound 5 of the present invention, when combined with the MEK inhibitor Trametinib, significantly enhanced the tumor inhibition rate against HPAF-II (KRAS G12D) xenografts in nude mice. The tumor inhibition rates of Compound 5 (50 mg / kg, bid) monotherapy, Trametinib (0.125 mg / kg, bid) monotherapy, and the combination therapy were 61.3%, 76.2%, and 105.3%, respectively (P < 0.001). In summary, in mouse models, the activity of compound 5 of the present invention as a single drug was roughly equivalent to that of BI-3402, and its combination with MRTX849 or Trametinib showed better anti-tumor activity against KRAS-mutant tumors, which was significantly better than the individual single-drug groups.

[0904] The applicant declares that this invention illustrates a pyrimidine-pyridone derivative as an SOS1 inhibitor, its preparation method, and its application through the above embodiments. However, this invention is not limited to the above embodiments, meaning that this invention does not necessarily rely on the above embodiments for implementation. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials, additions of auxiliary components, and selection of specific methods, etc., all fall within the protection and disclosure scope of this invention.

[0905] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0906] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. Pyrimidine-pyridinone derivatives having the following structures: ； 。 2. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the pyrimidine-pyridone derivative of claim 1.

3. Use of the pyrimidopyridone derivative of claim 1 or the pharmaceutical composition of claim 2, alone or in combination with a KRAS inhibitor or a MEK signaling pathway inhibitor, in the preparation of a medicament for the treatment of cancer or in the preparation of an SOS1 inhibitor.

4. The use according to claim 3, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and lung cancer, and the KRAS inhibitor is a KRASG12C, KRASG12V, KRASG12S, or KRASG12D inhibitor; The MEK signaling pathway inhibitor is a RAF, MEK, or ERK1 / 2 inhibitor.

Images