WO2022170802A1

WO2022170802A1 - Pyrimido-pyridone derivative as sos1 inhibitor, preparation method therefor and use thereof

Info

Publication number: WO2022170802A1
Application number: PCT/CN2021/131012
Authority: WO
Inventors: 谢成英; 郑苏欣; 陆晓杰; 郑明月; 乔刚; 叶阳亮
Original assignee: 苏州阿尔脉生物科技有限公司
Priority date: 2021-02-09
Filing date: 2021-11-16
Publication date: 2022-08-18
Also published as: CN116669738A

Abstract

A pyrimido-pyridone derivative as an SOS1 inhibitor, a preparation method therefor and a use thereof. The pyrimido-pyridone derivative has a structure as shown in formula (I), and has significant effects for inhibiting an RAS signal pathway and treating and/or preventing cancers. The cancers comprise pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal carcinoma, gastric cancer, cholangiocarcinoma and the like.

Description

A kind of pyrimidopyridone derivative as SOS1 inhibitor, its preparation method and use

technical field

The invention belongs to the technical field of pyrimidopyridone derivatives, in particular to a pyrimidopyridone derivative as an SOS1 inhibitor, a preparation method and uses thereof.

Background technique

The RAS family proteins belong to a small GTPase that includes three subfamilies, KRAS, NRAS, and HRAS. The mutated RAS gene is an important oncogene, and RAS gene mutations are found in 20-30% of human tumors, especially pancreatic, colorectal, and lung cancers. Various isoforms of RAS proteins have a balance between the activated state of GTP binding and the inactive state of GDP binding. GTPase-activating proteins (GAPs) can promote the conversion of GTP to GDP, thereby making RAS proteins inactive. The active state transition, and guanine nucleotide exchange factors (guanine nucleotide exchange factors, GEFs) can promote the release of GDP and the binding of GTP, thereby turning the RAS protein into the active state. Activation of RAS protein can promote cell proliferation, apoptosis evasion and metabolic reorganization through the RAS-RAF-MEK-ERK and RAS-PI3K-PDK1-AKT signaling pathways, thereby promoting the occurrence and development of tumors.

SOS1 (son of sevenless 1) is a key guanine nucleotide exchange factor (GEF) that can bind to RAS protein, promote the binding of RAS protein to GTP, and turn RAS protein into an active state. Recent studies have found that SOS1 inhibitors alone can inhibit the growth of various RAS mutant cells, and can also synergize with MEK inhibitors, resulting in significant inhibition of KRAS-driven tumors ^1-2 . The development of SOS1 inhibitors has become a research hotspot. Several patents have reported SOS1 inhibitors of different structural types, such as WO2018172250, WO2019201848, WO2018115380, WO2019122129, WO2020173935, WO2020180768 and WO2020180770, etc.

However, there are still uncertainties in the efficacy, safety or selectivity of the compounds disclosed in the prior art and the experimental drugs, so it is necessary to research and develop new selective SOS1 inhibitors.

references:

1. Hillig et al. Discovery of poetent SOS1inhibitors that block RAS activation via disruption of the RAS-SOS1interaction.PNAS.116,2251-2560(2019).

2. Hofmann et al.BI-3406, a potent and selective SOS1::KRAS interaction inhibitor, is effective in KRAS-driven cancers through combined MEK inhibition. Cancer Discov. CD-20-0142 (2020).

SUMMARY OF THE INVENTION

In order to solve the above problems of the prior art, the object of the present invention is to provide a pyrimidopyridone derivative, its pharmaceutically acceptable salt, its tautomer or its stereoisomer, so as to screen out the Compounds with excellent properties in terms of efficacy, safety and selectivity as SOS1 inhibitors.

Another object of the present invention is to provide a method for the preparation of the derivatives, their pharmaceutically acceptable salts, their tautomers or their stereoisomers.

In order to achieve this object of the invention, the present invention adopts the following technical solutions:

In a first aspect, the present invention provides a pyrimidopyridone derivative, a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, wherein the structure of the pyrimidopyridone derivative is as follows Formula (I) shows:

Wherein: R ¹ is selected from hydrogen or C ₁ -C ₃ alkyl; preferably hydrogen or methyl;

R ² is selected from hydrogen or C ₁ -C ₃ alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein C ₁ -C ₃ alkyl, 3-7 membered cycloalkyl, 4-7 membered cycloalkyl A membered heterocyclyl group is optionally substituted with 1-3 R ²¹ ;

R ²¹ is selected from C ₁ -C ₃ alkyl, hydroxy, halogen, cyano, amino, C ₁ -C ₃ alkoxy or =O;

L may be absent or selected from O, NH or N-(C ₁ -C ₃ alkyl);

R ³ is selected from H, C ₁ -C ₃ alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein C ₁ -C ₃ alkyl, 3-7 membered cycloalkyl, 4-7 membered cycloalkyl A membered heterocyclyl group is optionally substituted with 1-3 R ³¹ ;

R ³¹ is selected from C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, hydroxy, halogen, cyano, -NR ^a R ^b , C ₁ -C ₃ alkoxy, =O, -NHCOR ³² or - COR ³² ;

R ^a is selected from H, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl or 3-6 membered cycloalkyl;

R ^b is selected from H, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl or 3-6 membered cycloalkyl;

R ³² is selected from C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclyl;

AR is selected from 6-10-membered aryl or 5-10-membered heteroaryl, wherein the aryl or heteroaryl is optionally substituted by 1-4 R ⁴ ;

R ⁴ is selected from H, halogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, hydroxy-C ₁ -C ₃ alkyl, hydroxy-C ₁ -C ₃ haloalkyl, 3-6 membered cycloalkane base, 4-7 membered heterocyclic group, -OR ^a , -NR ^a R ^b , 6-10 membered aryl group or 5-10 membered heteroaryl group, of which 6-10 membered aryl group or 5-10 membered aryl group The heteroaryl group is optionally substituted by 1-4 R ^c ;

R ^c is selected from H, halogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, hydroxy-C ₁ -C ₃ alkyl, hydroxy-C ₁ -C ₃ haloalkyl, 3-6 membered cycloalkane base, 4-7 membered heterocyclic group, -OR ^a , -NR ^a R ^b , NR ^a R ^b -C ₁ -C ₄ alkyl, NR ^a R ^b -C ₁ -C ₄ haloalkyl; the formula ( The heteroatoms in the heterocyclic group or heteroaryl group in I) are 1-3 and are selected from one or more of oxygen, nitrogen and sulfur.

Preferably, the structure of the pyrimidopyridone derivatives is shown in formula (II):

wherein, R ¹ , R ² , R ³ and R ⁴ have the same defined ranges as above; n=1-4 (eg n=1, n=2, n=3, n=4);

Preferably, in the compound of formula (II), the phenyl group is optionally substituted by 1-4 R ⁴ , when the number of the R ⁴ is 2-4, the R ⁴ can be the same or different;

And/or, when the R ⁴ is a C ₁ -C ₃ haloalkyl group, the R ⁴ is 1-3, and when there are two or more R ⁴ , the R ⁴ can be the same or different;

And/or, when the R ⁴ is a C ₁ -C ₃ haloalkyl group, the halogen atom therein is fluorine;

And/or, when the R ⁴ is halogen, the R ⁴ is 1-3, and when there are two or more R ⁴ , the R ⁴ can be the same or different;

And/or, when the R ⁴ is halogen, the halogen atom is fluorine;

And/or, when the R ⁴ is -NR ^a R ^b , the R ⁴ is 1-3, and when there are two or more R ⁴ , the R ⁴ can be the same or different;

And/or, when the R ⁴ is -NR ^a R ^b , the R ^a and R ^b can be the same or different;

and/or R ³ is H, C ₁ -C ₃ alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein the 4-7 membered heterocyclyl is optionally surrounded by 1-3 R ³¹ replaced;

And/or, when R ³ is a 4-7-membered heterocyclic group optionally substituted by 1-3 R ³¹ , and when said R ³¹ is 2-3, R ³¹ is the same or different;

And/or, when the R ³ is a 4-7-membered heterocyclic group optionally substituted by 1-3 R ³¹ , the heterocyclic group contains 1-2 heteroatoms;

And/or, when the R ³ is a 4-7 membered heterocyclic group optionally substituted by 1-3 R ³¹ , the heteroatom of the heterocyclic group is nitrogen and/or oxygen;

And/or, when the R ³ is a 4-7-membered heterocyclic group optionally substituted by 1-3 R ³¹ , when the heteroatoms of the heterocyclic group are two, the two hetero atoms are the same or different;

And/or, when the R ³ is a 4-7-membered heterocyclic group optionally substituted by 1-3 R ³¹ , the R ³¹ is selected from C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, hydroxy, halogen, cyano, -NR ^a R ^b , C ₁ -C ₃ alkoxy, =O, -NHCOR ³² or -COR ³² ;

R ³² is selected from C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclyl.

Preferably, the structure of the pyrimidopyridone derivatives is shown in formula (III):

Wherein, R ¹ , R ² , R ³ and R ⁴ have the same defined ranges as above; n=1-4 (eg n=1, n=2, n=3, n=4).

Further preferably, for the compound of formula (III), the phenyl group is optionally substituted by 1-4 R ⁴ , when the number of the R ⁴ is 2-4, the R ⁴ can be the same or different;

And/or, when the R ⁴ is halogen, the R ⁴ is 1-2, and when there are more than two R ⁴ , the R ⁴ can be the same or different;

And/or, when the R ⁴ is halogen, the halogen atom is fluorine;

And/or, when the R ⁴ is -NR ^a R ^b , the R ⁴ is 1-3, and when there are more than two R ⁴ , the R ⁴ can be the same or different;

Preferably, the structure of the pyrimidopyridone derivatives is shown in formula (IV):

Wherein, R ¹ , R ² , R ³ , R ^c and L have the same defined ranges as above; m=1-4 (eg m=1, m=2, m=3, m=4). Further preferably, in the compound of formula (IV), L is selected from O, NH or NCH ₃ .

Further preferably, the pyrimidopyridone derivatives are selected from any one of the following structures:

In the second aspect, the present invention provides a preparation method of the pyrimidopyridone derivative, its pharmaceutically acceptable salt, its tautomer or its stereoisomer as described in the first aspect, which is selected from from one of the following two options:

Option One

The preparation method of the compound described in the general formula (I) of the present invention or its stereoisomer, tautomer or its pharmaceutically acceptable salt, comprises the following steps:

Preparation of key intermediate (I-A):

In the first step, the aromatic compound of general formula (I-1) is subjected to metal-catalyzed cross-coupling to obtain the compound of general formula (I-2);

In the second step, the compound of the general formula (I-2) is reacted under catalyst conditions to obtain a chiral sulfonimide compound of the general formula (I-3);

In the third step, the compound of general formula (I-3) is reduced by a metal reducing agent to obtain a chiral compound of general formula (I-4);

In the fourth step, the compound of general formula (I-4) is cleaved by sulfonamide under acidic conditions to obtain a chiral benzylamine compound of general formula (I-A).

The preparation of key intermediate (I-B), method one:

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

In the second step, the compound of general formula (I-6) is removed tert-butyl under acidic conditions to obtain the compound of general formula (I-7);

The 3rd step, the compound of general formula (I-7) obtains the compound of general formula (I-8) through esterification reaction with alcohol under acidic conditions;

In the fourth step, the compound of the general formula (I-9) and the amine of the general formula (I-10) are subjected to substitution reaction to obtain the compound of the general formula (I-11);

The 5th step, the compound of general formula (I-11) obtains the compound of general formula (I-12) through halogenation reaction;

The 6th step, the compound of general formula (I-12) and boronic acid or boron ester compound obtain the compound of general formula (I-13) through Suzuki reaction under basic conditions, in the presence of metal catalyst and ligand;

The seventh step, double bond oxidation in the compound of general formula (I-13) obtains the compound of general formula (I-14);

In the eighth step, the compound of general formula (I-14) and the compound of general formula (I-8) are obtained under basic conditions to obtain the compound of general formula (I-B).

The preparation of key intermediate (I-B), method two:

In the first step, the compound of the general formula (I-15) and the amine of the general formula (I-10) are subjected to substitution reaction to obtain the compound of the general formula (I-16);

In the second step, the compound of the general formula (I-16) and the compound of the general formula (I-8) are obtained under basic conditions to obtain the compound of the general formula (I-B).

Preparation of general formula (I):

In the first step, the compound of general formula (I-B) and the compound of general formula (I-A) are subjected to substitution reaction to obtain the compound of general formula (I).

Wherein, X, X ¹ , X ² and X ³ are halogen; X is preferably bromine; X ¹ and X ² are preferably chlorine; X ³ is preferably iodine; R ¹ , R ² , R ³ , AR and L have the same the same limits.

Option II

In the first step, the compound of general formula (I-12) and methyl acrylate are subjected to Heck reaction to obtain the compound of general formula (I-17) under the adjustment and reduction of basicity and metal catalyst;

In the second step, the compound of the general formula (I-17) is formed into an amide in the molecule under basic conditions to obtain the compound of the general formula (I-18);

In the third step, the compound of the general formula (I-18) is subjected to halogenation to obtain the compound of the general formula (I-19);

The 4th step, the compound of general formula (I-19) is demethylated under acidic conditions to obtain the compound of general formula (I-20);

The 5th step, the compound of general formula (I-20) and 2,4,6-triisopropylbenzenesulfonyl chloride or phosphorus oxychloride react under basic conditions to obtain the compound of general formula (I-21);

In the sixth step, the compound of general formula (I-21) and the compound of general formula (I-A) are reacted under basic conditions to obtain the compound of general formula (I-22);

The seventh step, the compound of general formula (I-22) and the compound of general formula (I-23) are obtained by Suzuki reaction under basic conditions, in the presence of a metal catalyst and a ligand, to obtain the compound of general formula (I) .

Wherein, X ¹ , X ³ and X ⁴ are halogen; X ¹ is preferably chlorine; X ³ is preferably iodine; X ⁴ is preferably bromine; X ⁵ is halogen or

W is

R ¹ , R ² , R ³ , AR and L have the same defined ranges as above.

For the above preparation method,

The reagents that provide alkaline conditions are selected from organic bases or inorganic bases, and the organic bases are triethylamine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, bismuth One or more of lithium trimethylsilylamide, sodium tert-butoxide, sodium methoxide and potassium tert-butoxide, and the inorganic bases are sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate , one or more of cesium carbonate, sodium hydroxide, potassium hydroxide, sodium bicarbonate and lithium hydroxide;

The reagent for providing acidic conditions is one or more of hydrogen chloride, hydrogen chloride solution in 1,4-dioxane, hydrogen chloride solution in methanol, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid and phosphoric acid kind;

The metal catalyst is palladium/carbon, Raney nickel, tetrakis-triphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride ( Pd(dppf)Cl ₂ ), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, bistriphenylphosphonium palladium dichloride (Pd(PPh) ₃ ) one or more of Cl ₂ ) and tris(dibenzylideneacetone)dipalladium (Pd ₂ (dba) ₃ );

The ligands are 2-dicyclohexylphosphine-2,6'-dimethoxybiphenyl (SPhos), 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (XantPhos), 2- Dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (XPhos), 2-dicyclohexylphosphino-2'-(N,N-dimethylamine)-biphenyl (DavePhos), 1, One or more of 1'-bis(diphenylphosphino)ferrocene (Dppf) and 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (BINAP), preferably 1,1 '-Binaphthalene-2,2'-bisdiphenylphosphine (BINAP);

The reducing agent is one or more of sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, lithium aluminum tetrahydrogen;

The oxidant is one or more of potassium permanganate, manganese dioxide, potassium dichromate, sodium dichromate and potassium osmate;

The above reaction is preferably carried out in a solvent, and the solvent used is N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, 1,4-dioxane, water, tetrahydrofuran, dichloromethane, One or more of 1,2-dichloroethane, methanol, ethanol, toluene, petroleum ether, ethyl acetate, n-hexane and acetone.

In a third aspect, the present invention provides a pharmaceutical composition comprising the pyrimidopyridone derivatives and/or stereoisomers, tautomers, pharmaceutically acceptable pyrimidopyridone derivatives as described in the first aspect salts; and pharmaceutically acceptable carriers and/or excipients and/or sustained release agents.

In the present invention, the term "comprising" means that the various ingredients can be used together in the mixture or composition of the present invention. Thus, the terms "consisting essentially of" and "consisting of" are encompassed by the term "comprising".

In the present invention, a "pharmaceutically acceptable" ingredient is one that is suitable for use in humans and/or animals without undue adverse side effects (eg, toxicity, irritation, and allergy), ie, with a reasonable benefit/risk ratio.

In the present invention, "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or excipient for delivering the active substance of the present invention or a physiologically acceptable salt thereof to animals or humans. The carrier can be liquid or solid.

In the present invention, the pharmaceutical composition contains a safe and effective amount (such as 0.001-99.9 parts by weight, more preferably 0.01-99 parts by weight, more preferably 0.1-90 parts by weight) of the compound represented by 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.

Or, 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 total weight of the compound represented by formula (I) or its pharmacy an acceptable salt; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100% by weight.

In another preferred example, the preferred ratio of the compound of formula (I) to a pharmaceutically acceptable carrier, excipient or sustained-release agent is that formula (I) as an active ingredient accounts for more than 65% of the total weight, and the rest accounts for The total weight ratio is 0.5-40%, or more preferably 1-20%, or more preferably 1-10%.

The various formulation forms of the pharmaceutical composition of the present invention, the unit dose of each dose comprises 0.05mg-500mg, preferably 0.5mg-200mg, more preferably 0.1mg-100mg of the compound of formula (I), enantiomer, external Racemates, pharmaceutically acceptable salts or mixtures thereof.

When the pharmaceutical composition contains additional active pharmaceutical ingredients for treating or preventing cancer, the amount of the active ingredients can generally be the conventional amount or lower in the prior art.

The pharmaceutical compositions of the present invention may be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, etc., wherein the compound of formula (I) may be present in a suitable solid or liquid carrier or diluent middle. The pharmaceutical compositions of the present invention may also be stored in a suitable sterile device for injection or instillation. The pharmaceutical composition may also contain odorants, flavoring agents, and the like.

The compound of formula (I) or the pharmaceutical composition comprising the compound of formula (I) of the present invention can be clinically used in mammals (including humans) by oral, nasal, dermal, pulmonary or gastrointestinal routes of administration. The preferred route of administration is oral. The preferred daily dose is 0.5 mg to 200 mg/kg body weight, taken in one or divided doses. Regardless of the method of administration, the optimal dose for an individual should depend on the specific treatment. It is common to start with a small dose and gradually increase the dose until the most suitable dose is found.

The effective dose of active ingredient employed may vary with the compound employed, the mode of administration, and the severity of the disease to be treated. Generally, however, satisfactory results are obtained when the compounds of the present invention are administered in doses of about 1-300 mg/kg of animal body weight per day, preferably in 1-3 divided doses per day, or in sustained release form Dosing. For most large mammals, the total daily dose is about 5-1000 mg, preferably about 10-500 mg. Dosage forms suitable for oral administration contain about 1-200 mg of the active compound in intimate admixture with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen can be adjusted to provide optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced, as dictated by the exigencies of the therapeutic situation.

The compounds or pharmaceutically acceptable salts and compositions thereof can 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, especially tablets and solid- or liquid-filled capsules. Oral administration of the pharmaceutical composition is preferred.

Solid carriers include: starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include: sterile water, polyethylene glycols, nonionic surfactants and edible oils (eg corn oil, peanut oil and sesame oil) as appropriate to the characteristics of the active ingredient and the particular mode of administration desired. Adjuvants commonly used in the preparation of pharmaceutical compositions may also advantageously be included, such as flavors, colors, preservatives and antioxidants such as vitamin E, vitamin C, BHT and BHA.

The active compounds or pharmaceutically acceptable salts thereof and compositions thereof may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds (as the free base or as a pharmaceutically acceptable salt) can also be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquids, polyethylene glycols, and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

The pharmaceutical forms suitable for injection include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, these forms must be sterile and must be fluid for easy syringe expelling. It must be stable under the conditions of manufacture and storage and must be resistant to the contaminating influence of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, alcohol (for example, glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

The compound represented by formula (I) or its pharmaceutically acceptable salt and its composition can also be administered in combination with other active ingredients or drugs for treating or preventing chronic pain diseases. When two or more drugs are administered in combination, they generally have better effects than when the two drugs are administered separately.

In the fourth aspect, the present invention provides a pyrimidopyridone derivative as described in the first aspect, its stereoisomer, tautomer, pharmaceutically acceptable salt or the pharmaceutical combination as described in the third aspect Use of the drug alone or in combination with a KRAS inhibitor in the preparation of a medicament for the treatment of cancer or in the preparation of an SOS1 inhibitor; preferably, the cancer includes but is 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, kidney cancer and bile duct cancer; further preferred Preferably, the cancer includes, but is not limited to: pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer and bile duct cancer; and/or the KRAS inhibitor is KRAS G12C, KRAS G12V, KRAS G12S or KRAS G12D inhibitor; and/or said MEK signaling pathway inhibitor such as RAF, MEK or ERK1/2 inhibitor.

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

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

The use according to the fourth aspect, wherein the cancer is a RAS family related cancer; preferably a KRAS, HRAS or NRAS G12C related cancer, KRAS, HRAS or NRAS G12D related cancer, KRAS, HRAS or NRAS G12V related cancer, KRAS, HRAS or NRAS G12S-related cancer, KRAS, HRAS or NRAS G12A-related cancer, KRAS, HRAS or NRAS G13D-related cancer, KRAS, HRAS or NRAS G13C-related cancer, KRAS, HRAS or NRAS Q61L-related cancer, KRAS, HRAS or NRAS A146T-related cancer, KRAS, HRAS or NRAS A146V-associated cancer or KRAS, HRAS or NRAS A146P-associated cancer.

In another aspect, there is provided a method of treating cancer comprising providing to a cancer patient a therapeutically effective amount of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof. In one embodiment, the cancer is a RAS family associated cancer. In one embodiment, the cancer is a SOS-L associated cancer. In one embodiment, the cancer is an NF-1/NF-2 related cancer.

The compositions and methods provided by the present invention can be used to treat a variety of cancers, including prostate, breast, brain, skin, cervical cancer, testicular cancer, and the like. More specifically, cancers treatable by the compositions and methods of the present invention include, but are not limited to, tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, cephalic Neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid cancers and sarcomas. More specifically, these compounds are useful in the treatment of: Heart: sarcomas (angiosarcomas, fibrosarcomas, rhabdomyosarcomas, liposarcoma), myxomas, rhabdomyomas, fibroids, lipomas and teratomas; lung: bronchial carcinoma (squamous 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 (cancer, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagon, gastrinoma, carcinoid, VIPoma), small intestine (glandular Carcinoma, lymphoma, carcinoid, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colorectal (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, smooth muscle) Genitourinary tract: kidney (adenocarcinoma, Wilms tumor (Wilms tumor), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma) , sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); liver : Liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, hemangiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone: osteosarcoma (osteosarcoma), fibrous Sarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, chronic bone tumor (chondrosarcoma), benign tumor , chondroblastoma, chondromyxoma, osteoid osteoma, and giant cell tumor; nervous system: skull (osteomas, hemangiomas, granulomas, xanthoma, osteitis), meninges (meningiomas, meningiosarcoma, glue plasmoma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal tumor), glioblastoma pleomorphism, oligodendroglioma , schwannoma, retinoblastoma, congenital tumors), spinal fibroma, meningioma, glioma, sarcoma; Gynecology: uterus (endometrial cancer), cervix (cervical cancer, precancerous cervical dysplasia) , Ovarian (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa sheath cell tumor, Sertolioma, germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, epithelial Internal carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, cystosarcoid (embryonic rhabdomyosarcoma), fallopian tube (carcinoma); hematology: blood (acute and chronic myeloid leukemia) , acute lymphocytic 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: neuroblastoma. In certain embodiments, cancer is Diffuse large B-cell lymphoma (DLBCL).

In one embodiment, the cancer is a RAS family related cancer, such as a KRAS, NRAS or HRAS related cancer. In certain embodiments, the RAS family-related cancer is non-small cell lung cancer or pancreatic cancer. In one embodiment, the cancer is a cancer associated with SOS 1. In certain embodiments, the SOS 1-related cancer is lung adenocarcinoma, embryonal rhabdomyosarcoma, Sertoli cell testicular tumor, and granulosa cell tumor of the skin. In one embodiment, the cancer is an NF-1 related cancer.

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

Terminology Explanation

Unless stated to the contrary, some terms used in the specification and claims of the present invention are defined as follows:

As used herein, "KRAS G12C" refers to a mutated form of the mammalian KRAS protein containing the amino acid at amino acid position 12 replaced by cysteine for glycine.

The determination of amino acid codons and residue positions of human KRAS was based on the amino acid sequence of UniProtKB/SwissProt P 01116: variant p.Gly 12 Cys.

"KRAS G12D" refers to a mutant form of the mammalian KRAS protein containing an amino acid with aspartic acid substituted for glycine at position 12. The amino acid codons and residue positions of human KRAS were determined from the amino acid sequence determined by UniProtKB/Swi-prot P01116: Variant P.Gly 12 Asp.

"KRAS G12V" refers to a mutant form of mammalian KRAS protein containing the amino acid valine in place of glycine at position 12. The amino acid codons and residue positions of human KRAS were determined according to the amino acid sequence determined by UniProtKB/Swi-prot P01116: Variant P.Gly 12 Val.

"KRAS G12S" refers to a mutant form of a mammalian KRAS protein containing the amino acid at amino acid position 12 with serine substituted for glycine. The amino acid codons and residue positions of human KRAS were determined from the amino acid sequence identified by UniProtKB/SwissProt P 01116: variant p.Gly 12Ser.

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

"KRAS G13D" refers to a mutant form of a mammalian KRAS protein containing the amino acid at amino acid position 13 aspartic acid in place of glycine. The determination of amino acid codons and residue positions of human KRAS is based on the amino acid sequence determined by UniProtKB/SwissProt P01116: variant p.gly 13Asp.

"KRAS G13C" refers to a mutant form of the mammalian KRAS protein containing the amino acid at amino acid 13 replaced by cysteine for glycine. The amino acid codons and residue positions of human KRAS were determined according to the amino acid sequence determined by UniProtKB/SwissProt-Prot P01116: Variant P.Gly 13Cys.

As used herein, "KRAS Q61L" refers to a mutant form of the mammalian KRAS protein containing the amino acid leucine in place of glutamine at amino acid 61. The determination of amino acid codons and residue positions of human KRAS is based on the amino acid sequence of UniProtKB/SwissProt P 01116: variant p.Gln61Leu.

As used herein, "KRAS A146T" refers to a mutant form of the mammalian KRAS protein containing the amino acid at amino acid position 146 with threonine in place of alanine. The amino acid codons and residue positions of human KRAS were determined from the amino acid sequence identified by UniProtKB/SwissProt P 01116: variant p.al46Thr.

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

As used herein, "KRAS A146P" refers to a mutated form of the mammalian KRAS protein containing the amino acid at amino acid position 146 in which a proline is substituted for alanine. The amino acid codons and residue positions of human KRAS were determined from the amino acid sequence identified in UniProtKB/SwissProt P 01116: variant p.Al46Pro.

As used herein, "HRAS G12C" refers to a mutated form of a mammalian HRAS protein containing the amino acid at amino acid position 12 replaced by cysteine for glycine. Based on the amino acid sequence identified in UniProtKB/SwissProt P 01112: variant p.Gly 12 Cys, the amino acid codons and residue positions of human HRAs were determined.

"HRAS G12D" refers to a mutant of a mammalian HRAS protein containing aspartic acid in place of glycine at amino acid position 12. The amino acid codons and residue positions of human HRAs were determined based on UniProtKB/SwissProt P01112: Amino acid sequence of variant P.Gly 12Asp.

"HRAS G12S" refers to a mutant form of a mammalian HRAS protein containing 12 amino acids with serines substituted for glycines. The amino acid codons and residue positions of human HRAS were determined according to the amino acid sequence determined by UniProtKB/SwissProt P01112: Variant P.Gly 12Ser.

"HRAS G12A" refers to a mutant form of a mammalian HRAS protein that replaces the amino acid at 12 with alanine for glycine. The amino acid codons and residue positions of human KRAS were determined from the amino acid sequence identified in UniProtKB/SwissProt P01112: variant P.Gly 12Ala.

"HRAS G13D" refers to a mutant form of a mammalian HRAS protein containing an amino acid with aspartic acid substituted for glycine at position 13. The amino acid codons and residue positions of human HRAS were determined according to the amino acid sequence determined by UniProtKB/Swi-prot P01112: variant p.gly 13Asp.

"HRAS G13C" is a mutant of mammalian HRAS protein, which contains cysteine in place of the amino acid of glycine at the 13th amino acid. The determination of amino acid codons and residue positions of human HRAS was based on the amino acid sequence of UniProtKB/SwissProt-Prot P01112: variant P.Gly 13Cys.

As used herein, "HRAS Q61L" refers to a mutated form of a mammalian HRAS protein containing the amino acid at amino acid position 61 in place of leucine in place of glutamine. Based on the amino acid sequence identified by UniProtKB/SwissProt P 01112 variant p.Gln61Leu, the amino acid codons and residue positions of human HRAs were determined.

As used herein, "HRAS A146T" refers to a mutated form of a mammalian HRAS protein containing the amino acid at amino acid position 146 with a threonine substituted for alanine. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified in UniProtKB/SwissProt P 01112: variant p.Al46Thr.

As used herein, "HRAS A146V" refers to a mutant form of a mammalian HRAS protein containing the amino acid valine at amino acid position 146 in place of alanine. One amino acid replaces the alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified in UniProtKB/SwissProt P 01112: variant p.al46Val.

As used herein, "HRAS A146P" refers to a mutated form of the mammalian HRAS protein containing the amino acid at amino acid position 146 where a proline is substituted for alanine. Amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified in UniProtKB/SwissProt P 01112: variant p.Al46Pro.

"NRAS G12C" refers to a mutant form of the mammalian NRAS protein containing the amino acid glycine at amino acid 12 replaced by cysteine. The amino acid codons and residue positions of human NRAS were determined according to UniProtKB/SwissProt-Prot P01111: variant P.Gly 12Cys.

"NRAS G12D" refers to a mutant form of the mammalian NRAS protein that contains an amino acid with aspartic acid substituted for glycine at position 12. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence determined by UniProtKB/Swi-prot P01111: Variant P.Gly 12 Asp.

"NRAS G12S" refers to a mutant form of the mammalian NRAS protein containing the amino acid serine substituted for glycine at amino acid 12. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified by UniProtKB/SwissProt P01111: variant p.Gly 12Ser.

"NRAS G12A" refers to a mutant form of the mammalian NRAS protein containing the amino acid at amino acid 12 where alanine is substituted for glycine. The determination of amino acid codons and residue positions of human KRAS was based on the amino acid sequence of UniProtKB/SwissProt P01111: variant p.Gly 12Ala.

As used herein, "NRAS G13D" refers to a mutated form of the mammalian NRAS protein that contains an amino acid substitution of aspartic acid at amino acid 13 for glycine at the amino acid position. The determination of amino acid codons and residue positions of human NRAS is based on the amino acid sequence determined by UniProtKB/SwissProt P 01111: variant p.Gly 13 Asp.

"HNRas G13C" refers to a mutant form of the mammalian NRAS protein containing the amino acid cysteine in place of glycine at amino acid 13. The determination of amino acid codons and residue positions of human NRAS was based on the amino acid sequence of UniProtKB/SwissProt P01111: variant P.Gly 13Cys.

As used herein, "HRAS Q61L" refers to a mutated form of a mammalian HRAS protein containing a leucine at amino acid position 61 in place of glutamine at amino acid position 61. Based on the amino acid sequence identified by UniProtKB/SwissProt P 01112 variant p.Gln61Leu, the amino acid codons and residue positions of human HRAs were determined.

As used herein, "NRAS A146T" refers to a mutant form of the mammalian NRAS protein containing the amino acid threonine at amino acid 146 in place of alanine. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified in UniProtKB/SwissProt P 01111: variant p.Al46Thr.

As used herein, "NRAS A146V" refers to a mutant form of a mammalian NRAS protein containing the amino acid valine at amino acid 146 in place of alanine. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified by UniProtKB/SwissProt P 01111: variant p.al46Val.

As used herein, "NRAS A146P" refers to a mutated form of the mammalian NRAS protein containing an amino acid with a proline in place of alanine at amino acid 146. The amino acid codons and residue positions of human NRAS were determined from the amino acid sequence identified by UniProtKB/SwissProt P 01111: variant p.Al46Pro.

As used herein, "RAS family member" or "RAS family" refers to KRAS, HRAS, NRAS and activating mutants thereof, including positions G12, G13, Q61 and A146.

As used herein, 'RAS family associated disease or disorder' refers to a disease or disorder associated with, mediated by, or having an activating RAS mutation, such as RAS at positions G12, G13, Q61 or A146. Non-limiting examples of RAS family related diseases or diseases are KRAS, HRAS or NRAS G12C related cancer, KRAS, HRAS or NRAS G12D related cancer, KRAS, HRAS or NRAS G12V related cancer, KRAS, HRAS or NRAS G12S related cancer, KRAS, HRAS or NRAS G12A-related cancer, KRAS, HRAS or NRAS G13D-related cancer, KRAS, HRAS or NRAS G13C-related cancer, KRAS, HRAS or NRAS Q61L-related cancer, KRAS, HRAS or NRAS A146T-related cancer, KRAS, HRAS or NRAS A146V-related cancer Cancer or KRAS, HRAS or NRAS 146P-related cancer.

As used herein, "SOS1-related disease or disorder" refers to a disease or disorder associated with, mediated by, or having an activating SOS1 mutation. Examples of activating SOS1 mutations include the SOS1N233S and SOS1N233Y mutations.

As used herein, "SOS1N233S" refers to a mutant form of the mammalian SOS1 protein containing an amino acid substitution of serine for glutamine at amino acid position 233. The determination of amino acid codons and residue positions for human SOS 1 was based on the amino acid sequence identified in UniProtKB/SwissProt Q 07889: variant p.Gln233Ser.

As used herein, "SOS1N233Y" refers to a mutant form of the mammalian SOS1 protein in which amino acid 233 is replaced by an amino acid containing tyrosine in place of glutamine. The determination of amino acid codons and residue positions of human SOS 1 was based on the amino acid sequence identified in UniProtKB/SwissProt Q 07889: variant p.Gln233Tyr.

As used herein, "SOS 1 inhibitor" refers to a compound of the present invention, as described herein, represented by formula (I). These compounds negatively inhibit all or part of the interaction of SOS 1 with RAS family mutants or SOS 1 activating mutations, thereby reducing and/or modulating the nucleotide exchange activity of the RAS family member-SOS 1 complex.

As used herein, "KRAS inhibitor" refers to inhibitors known in the art against various KRAS mutations. Inhibitors such as KRAS G12C, KRAS G12D, KRAS G12V can be targeted.

As used herein, "MEK signaling pathway" inhibitors refer to inhibitors of RAF, MEK and ERK1/2 known in the art.

"NF-1/NF-2-related disease or disorder" refers to a disease or disorder caused by a loss-of-function mutation in the neurocellulase (NF-1) gene or the neurocellulase 2 (NF-2) gene.

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), in-frame mutation or in-frame transfer mutation, including insertions and deletions, and the target cell Or homozygous deletion of a protein-encoding gene in a cancer cell, resulting in partial or complete loss of presence, activity, and/or function of the encoded protein.

"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 a saturated linear or branched monovalent hydrocarbon group of 1-2 carbon atoms, wherein the alkyl group may be independently optionally substituted by one or more substituents described herein. 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-di Methylbutyl etc. Alkyl groups can be optionally substituted or unsubstituted.

"Alkenyl" refers to a linear or branched monovalent hydrocarbon group of 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one CC is sp ² double bond, wherein the alkenyl group can be independently optionally substituted by one or more substituents described in the present invention, specific examples of which include, but are not limited to, vinyl, allyl and alkene Butyl and so on. Alkenyl groups can be optionally substituted or unsubstituted.

"Cycloalkyl" means a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring comprising 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl , cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups. Cycloalkyl groups can be optionally substituted or unsubstituted.

"Spirocycloalkyl" refers to a polycyclic group with 5 to 18 members, two or more cyclic structures, and the single rings share one carbon atom (called spiro atom) with each other, and the ring contains one or more aromatic systems with double bonds but none of the rings have fully conjugated pi electrons. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the rings, spirocycloalkyl groups are divided into mono-spiro, double-spiro or poly-spirocycloalkyl groups, preferably mono-spiro and double-spirocycloalkyl groups, preferably 4-membered/5-membered, 4-membered Yuan/6 Yuan, 5 Yuan/5 Yuan or 5 Yuan/6 Yuan. Non-limiting examples of "spirocycloalkyl" include, but are not limited to:

"Fused cycloalkyl" refers to a 5- to 18-membered all-carbon polycyclic group containing two or more cyclic structures that share a pair of carbon atoms with each other, and one or more rings may contain one or more double bonds, But none of the rings have an aromatic system with fully conjugated pi electrons, preferably 6 to 12 membered, more preferably 7 to 10 membered. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicycloalkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to:

"Bridged cycloalkyl" refers to an all-carbon polycyclic group of 5 to 18 members, containing two or more cyclic structures, sharing two carbon atoms that are not directly connected to each other, and one or more rings may contain one or more Aromatic systems in which multiple double bonds, but none of the rings have fully conjugated pi electrons, are preferably 6 to 12 membered, more preferably 7 to 10 membered. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to:

The cycloalkyl ring can be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring connected to the parent structure is a cycloalkyl, non-limiting examples include indanyl, tetrahydronaphthalene base, benzocycloheptyl, etc.

"Heterocyclyl", "heterocycle" or "heterocyclic" are used interchangeably in this application, and are used interchangeably in this application, and all refer to a saturated or partially unsaturated monocyclic ring containing 3-12 ring atoms , bicyclic or tricyclic non-aromatic heterocyclic groups, wherein at least one ring atom atom is a heteroatom, such as oxygen, nitrogen, sulfur atom and the like. It preferably has a 5- to 7-membered monocyclic ring or a 7- to 10-membered bi- or tricyclic ring, which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidine base, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and Piperazinyl. The heterocyclyl ring can be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl. Heterocyclyl groups can be optionally substituted or unsubstituted.

"Spiroheterocyclyl" refers to a polycyclic group with 5 to 18 members, two or more cyclic structures, and single rings share one atom with each other, and the ring contains one or more double bonds, but no An aromatic system with fully conjugated pi electrons in one ring, wherein one or more ring atoms are selected from nitrogen, oxygen, sulfur or S(O) _m heteroatoms, the remaining ring atoms are carbon, and m=1 or 2. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the rings, spiroheterocyclyls are classified into mono-spiroheterocyclyl, bis-spiroheterocyclyl or poly-spiroheterocyclyl, preferably mono-spiroheterocyclyl and bis-spiroheterocyclyl. 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 monospiroheterocyclyl group. Non-limiting examples of "spiroheterocyclyl" include, but are not limited to:

"Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more ring structures sharing a pair of atoms with each other, one or more rings may contain one or more double bonds, but no ring has complete Conjugated pi-electron aromatic systems in which one or more ring atoms are selected from nitrogen, oxygen, sulfur or S(O) _m heteroatoms, the remaining ring atoms are carbon, and m=1 or 2. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic groups. Non-limiting examples of "fused heterocyclyl" include, but are not limited to:

"Bridged heterocyclyl" refers to a 5- to 18-membered polycyclic group containing two or more cyclic structures that share two atoms that are not directly connected to each other, and one or more rings may contain one or more Double bonds, but none of the rings have a fully conjugated pi-electron aromatic system, where one or more ring atoms are selected from nitrogen, oxygen, sulfur, or heteroatoms of S(O) _m , the remaining ring atoms are carbon, and m = 1 or 2. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclyl" include, but are not limited to:

"Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be joined together in a fused fashion. The term "aryl" includes aromatic groups such as phenyl, naphthyl, tetrahydronaphthyl. Preferred aryl groups are _C6 - _C10 aryl groups, more preferred aryl groups are phenyl and naphthyl, and most preferred are phenyl groups. Aryl groups can be substituted or unsubstituted. The "aryl" can be fused with a heteroaryl, a heterocyclyl or a cycloalkyl, wherein the parent structure is linked together by an aryl ring, non-limiting examples include but are not limited to:

"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, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl , oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzo-diazolyl Oxolyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolinyl, indazolyl, benzisothiazolyl, benzo oxazolyl and benzisoxazolyl. Heteroaryl groups can be optionally substituted or unsubstituted. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples include but are not limited to:

"Alkoxy" refers to a group (alkyl-O-). Wherein, alkyl is as defined herein. Ci _- _C6 alkoxy groups are preferred. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

"Haloalkyl" refers to an alkyl group having one or more halogen substituents, wherein the alkyl group has the meaning as described herein. Examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, perfluoroethyl, 1,1-dichloroethyl, 1,2-dichloropropyl, and the like.

"Hydroxy" refers to the -OH group.

"Halogen" means fluorine, chlorine, bromine and iodine, preferably fluorine, chlorine and bromine.

"Amino" refers to _-NH2 .

"Cyano" refers to -CN.

"Nitro" refers to _-NO2 .

"Benzyl" refers to _-CH2 -phenyl.

"Carboxyl" refers to -C(O)OH.

"Acetyl" refers to -C(O) _CH3 or Ac.

"Carboxylate" means -C(O)O(alkyl) or (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

"Optional" means that the event it describes can, but need not, occur. For example, the description "AR is optionally substituted with 1 to more ^Rc " includes instances where the AR group may or may not be substituted with 1 to more ^Rc ^.

"Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of each other, are substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the person skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.

"Substituted" or "substituted" mentioned in this specification, unless otherwise specified, means that the group may be substituted by one or more groups selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy , alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Thio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =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) ₂ NR ^b R ^b , wherein R ^b is defined as in the general formula as described in (I).

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, cows, sheep, horses, primates animals and humans. In some embodiments, the patient is a human. In some embodiments, the subject has experienced and/or displayed at least one symptom of the disease or disease to be treated and/or prevented. In some embodiments, the subject has been identified or diagnosed with cancer with a KRAS G12 or G13 mutation (eg, as determined by an FDA-approved regulatory agency, such as an FDA-approved assay or kit). In some embodiments, the subject has a tumor that is positive for a KRAS G12C mutation, a KRAS G12D mutation, a KRAS G12S mutation, a KRAS G12V mutation, a KRAS G12A mutation, a KRAS G13D mutation, or a KRAS G13C mutation (eg, as approved by a regulatory agency-approved test or kit determination). The subject can be a person with a KRAS G12C mutation, a KRAS G12D mutation, a KRAS G12V mutation, a KRAS G12S mutation, a KRAS G12A mutation, a KRAS G13D mutation, or a KRAS G13C mutation (e.g., an approved regulatory agency-such as an FDA-approved assay or reagent) Box) positive tumor patients. The subject can be a subject whose tumor has a KRAS G12C mutation, a KRAS G12D mutation, a KRAS G12V mutation, a KRAS G12S mutation, a KRAS G12A mutation, a KRAS G13D mutation, or a KRAS G13C mutation (eg, the tumor was approved by an FDA-approved regulatory agency, kit or assay). In some embodiments, the subject is suspected of having a KRAS G12 or G13 gene-related cancer. In some embodiments, the subject has a clinical record indicating that the subject has a tumor with a KRAS G12C mutation (and optionally the clinical record indicating that the subject should be treated with any of the compositions provided herein).

The term "pediatric patient" as used herein refers to a patient under the age of 16 at the time of diagnosis or treatment. The term "child" can also be divided into the following subcategories: neonatal (from birth to first month of life); infant (1 month to two years); child (2 to 12 years); adolescent (12 years) Age to 21 (until but not including 22nd birthday). Berhman RE, Kliegman R, Arvin AM, Nelson WE. Nelson Textbook of Pediatrics, 15th ed. Philadelphia: W.B. Saunders Company, 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.

As used herein, an "effective amount" of a compound refers to an amount sufficient to negatively modulate or inhibit SOS 1 enzymatic activity.

As used herein, a "therapeutically effective dose" of a compound refers to an amount sufficient to ameliorate or somehow reduce symptoms, halt or reverse disease progression, or negatively modulate or inhibit SOS 1 activity. This dose can be given as a single dose or in a regimen to be effective.

As used herein, "treating" means in any way ameliorating or otherwise altering the symptoms or pathology of a patient's condition, disorder, or disease.

As used herein, "improving the symptoms of a particular disease by use of a particular compound or pharmaceutical composition" means any reduction, whether permanent or temporary, attributable to or associated with use of the composition Sexual, permanent or temporary.

The definitions and conventions of stereochemistry used in the present invention are generally referred to the following documents:

S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-HillBook Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York , 1994. The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomers. All stereoisomeric forms of the compounds of the present invention, including, but not limited to, diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, constitute the part. Diastereomers can be separated into individual diastereomers by methods such as chromatography, crystallization, distillation or sublimation on the basis of their physicochemical differences. Enantiomers can be separated by converting chiral mixtures into diastereomeric mixtures by reaction with an appropriate optically active compound such as a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride , the diastereomers are separated, and the individual diastereomers are converted to the corresponding pure enantiomers. The intermediates and compounds of the present invention may also exist in different tautomeric forms, and all such forms are included within the scope of the present invention. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D, L or R, S are used to denote the absolute configuration of the chiral center of the molecule. The prefixes d, l or (+), (-) are used to designate the sign of the plane-polarized light rotation of the compound, (-) or l means the compound is levorotatory, and the prefix (+) or d means the compound is dextrorotatory. The atoms or groups of atoms of these stereoisomers are connected to each other in the same order, but their steric structures are different. A specific stereoisomer may be an enantiomer, and a mixture of isomers is often referred to as an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, devoid of optical activity.

"Tautomer" or "tautomeric form" means that isomers of structures of different energies can be interconverted through a low energy barrier. For example, proton tautomers (ie, prototropic tautomers) include interconversions by migration of protons, such as keto-enol and imine-enamine isomerizations. Valence (valence) tautomers include interconversions that recombine bond electrons. Unless otherwise indicated, the structural formulas described herein include all isomeric forms (such as enantiomers, diastereomers, and geometric isomers): such as R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers. Accordingly, individual stereochemical isomers of the compounds of the present invention or mixtures of enantiomers, diastereomers, or geometric isomers thereof are within the scope of the present invention.

"Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention which are safe and effective in humans or animals. The salts of the compounds can be obtained by using a sufficient amount of base or acid in neat solution or in a suitable inert solution to obtain the corresponding addition salts. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts, etc. Pharmaceutically acceptable acid addition salts include inorganic acid salts and organic acid salts, and the inorganic and organic acids include 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, rice butenedioic acid, Phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, and methanesulfonic acid, among others (see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66:1-19 (1977)).

The present invention provides an SOS1 inhibitor with a new structure. The test results show that the pyrimidopyridone derivative exhibits excellent SOS1 inhibitory activity, and at the same time exhibits excellent safety and selectivity, and can be used to prepare and treat cancer. Especially drugs for diseases such as pancreatic cancer, colorectal cancer, lung cancer, hepatocellular cancer, kidney cancer, stomach cancer and bile duct cancer.

Description of drawings

Fig. 1 is a graph showing the effect of the compounds involved in the present invention on the KRAS/ERK1/2 signal transduction pathway of K-562 cells.

Figure 2 is a graph showing the in vivo antitumor effect of the SOS1 inhibitor of the present invention and MRTX849 alone or in combination on the KRASG12C mutant MIAPaCa-2 pancreatic xenograft nude mouse model.

Figure 3 is a graph showing the effect of the SOS1 inhibitor of the present invention and MRTX849 alone or in combination on the body weight of the KRASG12C mutant MIAPaCa-2 pancreatic xenograft nude mouse model.

Figure 4 is a graph showing the in vivo antitumor effect of the SOS1 inhibitor of the present invention and Trametinib alone or in combination on a KRASG12D mutant HPAF-II pancreatic xenograft tumor nude mouse model.

Figure 5 is a graph showing the effect of the SOS1 inhibitor of the present invention and Trametinib alone or in combination on the body weight of KRASG12D mutant HPAF-II pancreatic xenograft nude mice model.

Detailed ways

The method of the present invention will be described below through specific examples, so as to make the technical solution of the present invention easier to understand and grasp, but the present invention is not limited thereto. In the following examples, the ¹ H NMR spectrum was measured with a Bruker instrument (400 MHz), and the chemical shifts were expressed in ppm. A tetramethylsilane internal standard (0.00 ppm) was used. ¹ H NMR representation: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, br=broadened, dd=doublet of doublet, dt=triplet doublet of peaks. When coupling constants are provided, they are in Hz.

The mass spectrum was measured by LC/MS, and the ionization mode was ESI.

HPLC model: Agilent 1260, Thermo Fisher U3000; Column model: Waters xbrige C18 (4.6*150mm, 3.5μm); Mobile phase: A: ACN, B: Water (0.1% H ₃ PO ₄ ); Flow rate: 1.0mL/min; Gradient: 5%A for 1min, increase to 20%A within 4min, increase to 80%A within 8min, 80%A for 2min, back to 5%A within 0.1min; Wavelength: 220nm; Column Incubator: 35°C.

The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.2mm-0.3mm, and the specification used for TLC separation and purification products is 0.4mm -0.5mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

In the following examples, all temperatures are in degrees Celsius unless otherwise indicated, and various starting materials and reagents were obtained from commercially available or synthesized according to known methods without further purification. For direct use, unless otherwise specified, commercially available manufacturers include but are not limited to Sinopharm Group, Bailingwei Technology Co., Ltd., Tixiai (Shanghai) Chemical Industry Development Co., Ltd., Shanghai Bide Pharmaceutical Technology Co., Ltd. and Shanghai Merrill Chemical Technology Co., Ltd. Wait.

CD ₃ OD: Deuterated methanol

CDCl ₃ : deuterated chloroform

DMSO-d ₆ : deuterated dimethyl sulfoxide

Pd ₂ (dba) ₃ : tris(dibenzylideneacetone)dipalladium

Pd(dppf)Cl ₂ : [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

XantPhos: 4,5-Bisdiphenylphosphine-9,9-dimethylxanthene

XPhos: 2-dicyclohexylphosphorus-2,4,6-triisopropylbiphenyl

HATU: 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate

TLC: Thin Layer Chromatography

HPLC: High Performance Liquid Chromatography

purity: purity

&:and

Hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.

There is no special description in the examples, and the solution in the reaction refers to an aqueous solution.

There is no special description in the examples, and the reaction temperature is room temperature, which is 20°C-30°C.

The monitoring of the reaction progress in the embodiment adopts thin layer chromatography (TLC), the developing solvent used in the reaction, the eluent system of the column chromatography used for purifying the compound or the developing solvent system of the thin layer chromatography method includes: A: Petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane: ethyl acetate; the volume ratio of the solvent varies according to the polarity of the compound, and a small amount of acidic or basic reagent can also be added Adjustments such as acetic acid or triethylamine, etc.

Preparation of intermediates

Intermediate 1

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

The first step (S)-tetrahydrofuran-3-yl 4-nitrobenzoate IN-1b

Under nitrogen protection, (R)-3-hydroxytetrahydrofuran IN-1a (10.0 g, 0.113 mol) was dissolved in tetrahydrofuran (130 mL), and nitrobenzoic acid (18.9 g, 0.113 mol) and triphenylphosphine ( 35.6 g, 0.136 mol), cooled to 0 °C, and a solution of diisopropyl azodicarboxylate (27.8 g, 0.137 mol) in tetrahydrofuran (20 mL) was added dropwise, and the reaction was performed at room temperature for 2 hours. TLC showed that the reaction of the raw materials was completed. The reaction solution was added with water, 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 obtain the title compound IN-1b (29.8 g, crude product, containing a small amount of triphenylene) as a white solid. oxyphosphorus).

The second step (S)-3-hydroxytetrahydrofuran IN-1c

Compound IN-1b (29.8 g, crude product) was dissolved in ethanol (300 mL), then sodium carbonate (25.0 g, 0.236 mol) was added, heated to 85° C. for 2 hours, and the reaction was completed by TLC. The reaction solution was cooled to room temperature, filtered, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound IN-1c (8.1 g, two-step yield 81%) as an oil.

The third step (S)-tert-butyl 2-((tetrahydrofuran-3-yl)oxy)acetate IN-1d

Compound IN-1c (3.0 g, 34.05 mmol) was dissolved in toluene (40 mL), tetrabutylammonium bromide (3.0 g, 9.30 mmol) and aqueous sodium hydroxide solution (30 mL, 360 mmol, 12 mol/L) were added, and the mixture was stirred at room temperature Then, tert-butyl bromoacetate (9.9 g, 50.75 mmol) was added, and the mixture was stirred at 20° C. overnight. 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 subjected to silica gel column chromatography to obtain the oily title compound IN-1d (5.5 g, yield 80%).

¹ H NMR (400MHz, CDCl ₃ ) δ 4.22-4.25(m, 1H), 3.97(s, 2H), 3.80-3.95(m, 4H), 1.96-2.09(m, 2H), 1.48(s, 9H) ).

The fourth step (S)-2-((tetrahydrofuran-3-yl)oxy)acetic acid IN-1e

Compound IN-1d (5.5 g, 27.19 mmol) was dissolved in dichloromethane (15 mL), trifluoroacetic acid (30 mL) was added, heated to 45° C. to react for 2 hours, TLC showed that the reaction was basically complete. The reaction solution was concentrated (with dichloromethane 3 times) to give the title compound IN-1e (13.5 g, crude) as a pale yellow oil, which was used directly in the next step.

The fifth step (S)-2-((tetrahydrofuran-3-yl)oxy)ethyl acetate IN-1

Compound IN-1e (13.5 g, crude product) was dissolved in ethanol (100 mL), concentrated sulfuric acid (30 mL) was added at room temperature, and the reaction was heated to 85° C. for 5 hours. The reaction solution was cooled to room temperature, poured into ice brine, 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 silica gel column chromatography to obtain the title compound IN-1( 2.1 g, two-step yield 44%).

¹ H NMR (400MHz, CDCl ₃ ) δ 4.29-4.18(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).

Intermediate 2

(R)-(2-(5-(1-Aminoethyl)thiophen-3-yl)benzyl)(methyl)carbamate IN-2

The first step 1-(4-bromo-2-thiophene)ethanone IN-2b

2-Acetylthiophene 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 (dissolved in carbon tetrachloride (90 mL) was slowly added dropwise. 13.48 g, 84.36 mmol). After the addition was completed, the reaction was carried out at room temperature for 4 hours, and the reaction of the raw material 2-acetylthiophene was monitored by TLC. The reaction solution was poured into ice water, extracted with ethyl acetate (30 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to silica gel column chromatography to obtain the title compound IN-2b (12.0 g, yield) as a yellow oil. 74%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.58 (d, J=1.2 Hz, 1H), 7.58 (d, J=1.2 Hz, 1H), 2.54 (s, 3H).

The second step (R)-N-(1-(4-bromothiophen-2-yl)ethylene)-2-methylpropane-2-sulfinamide IN-2c

Tetraethyl titanate (60 mL) was added to the round-bottomed flask, and compound IN-2b (6.0 g, 29.26 mmol) and (R)-(+)-tert-butylsulfinamide (9.58 g, 79.04 mmol) were sequentially added at room temperature ), the temperature was slowly raised to 100 °C for 1 hour, and the reaction of the raw materials was monitored by TLC for completeness. The reaction solution was cooled to room temperature, poured into ice water, extracted with ethyl acetate (50 ml × 3 times), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to silica gel column chromatography to obtain the title compound IN-2c (8.6 g, yield 95%).

The third step (R)-N-((R)-1-(4-bromothiophen-2-yl)ethyl)-2-methylpropane-2-sulfinamide IN-2d

Compound IN-2c (6.4g, 20.76mmol) was dissolved in a mixed solvent of tetrahydrofuran (60mL) and water (1.2mL), cooled to -50°C, sodium borohydride (2.23g, 58.95mmol) was added in batches, and the addition was complete. , the reaction was slowly raised to room temperature for 2 hours, and the reaction of the raw materials was detected by TLC. The reaction solution was diluted with ethyl acetate, washed twice with saturated brine, twice with water, dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to silica gel column chromatography to obtain the title compound IN-2d (5.0 g, yield 78%).

The fourth step (R)-1-(4-bromothiophen-2-yl)-1-ethylamine IN-2e

To tetrahydrofuran (50 mL), compound IN-2d (5.0 g, 16.11 mmol) and concentrated hydrochloric acid (12 N, 0.5 mL) were added, and the temperature was raised to 80° C. and stirred for 1 hour. TLC detected that the reaction of the raw materials was complete. The reaction solution was cooled to room temperature, added with saturated aqueous sodium bicarbonate solution to adjust pH=8, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound IN-2e (3.34 g, crude product), used directly in the next step.

The fifth step (R)-(1-(4-bromothiophen-2-yl)ethyl) tert-butyl carbamate IN-2f

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

The sixth step (R)-(1-(4-(2-formylphenyl)thiophen-2-yl)ethyl)carbamic acid tert-butyl ester IN-2g

Under nitrogen protection, compound IN-2f (15.0 g, 48.98 mmol) and 2-formylbenzeneboronic acid (8.8 g, 58.78 mmol) were dissolved in 1,4-dioxane (100 mL) and water (10 mL), [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (600 mg, 0.735 mmol) and sodium carbonate (10.4 g, 98.12 mmol) were added at room temperature, and the temperature was raised to 90° C. to react for 3 hours. The reaction of the raw materials was basically completed by TLC. The reaction solution was cooled to room temperature, diluted with water, 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 obtain the title compound IN-2g (7.9 g, yield 49%).

The seventh step (R)-(1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)carbamic acid tert-butyl ester IN-2h

Compound IN-2g (7.9g, 23.84mmol) was dissolved in methylamine solution (25mL, 30% ethanol solution), stirred at room temperature overnight, added sodium cyanoborohydride (3.0g, 47.74mmol), stirred at room temperature for 2 hours, TLC The detection of the raw materials basically reacted completely. The reaction solution was quenched by adding water, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound IN-2h (7.5 g, crude product), which was directly used in the next step.

LC-MS: m/z=347.2[M+H] ⁺

The eighth step (R)-(2-(5-(1-((tert-butoxycarbonyl)amino)ethyl)thiophen-3-yl)benzyl)(methyl)carbamate IN-2i

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

The ninth step (R)-(2-(5-(1-aminoethyl)thiophen-3-yl)benzyl)(methyl)carbamate IN-2

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

¹ H NMR (400MHz, DMSO-d ₆ )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).

Intermediate 3

(R)-1-(3-Nitro-5-(trifluoromethyl)phenyl)ethanamine IN-3

The first step 1-(3-nitro-5-(trifluoromethyl)phenyl)ethan-1-one IN-3b

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

¹ H NMR (400MHz, CDCl ₃ ) δ 8.94(s,1H), 8.68(s,1H), 8.53(s,1H), 2.75(s,3H).

The second step (R,Z)-2-methyl-N-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethylene)propane-2-sulfinamide IN-3c

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 After 3 hours of reaction, TLC showed a small amount of starting material remaining. The reaction solution was cooled to room temperature, poured into ice water (60 mL), 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 silica gel column chromatography to obtain the title compound IN as a yellow oil. -3c (1.01 g, 56% yield).

The third step (R)-2-methyl-N-((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)propane-2-sulfinamide IN- 3d

Compound IN-3c (260 mg, 0.77 mmol) was dissolved in tetrahydrofuran (2.5 mL) and water (0.05 mL), cooled to -60°C, and sodium borohydride (74 mg, 1.95 mmol) was added in batches, and the addition was completed, keeping -60 Stirring was continued for 1 hour at °C, TLC indicated the reaction was complete. The reaction solution was quenched by dropwise addition of water, 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 silica gel column chromatography to obtain the title compound IN-3d (150 mg, yield 58) as a white solid %).

¹ H NMR (400MHz, CDCl ₃ ) 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).

The fourth step (R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethanamine IN-3

Compound IN-3d (164 mg, 0.48 mmol) was dissolved in tetrahydrofuran (3 mL), concentrated hydrochloric acid (0.5 mL) was added dropwise, and the reaction was completed at room temperature for 1 hour. TLC showed that the reaction was complete. The reaction was added dropwise with saturated aqueous sodium carbonate solution to adjust pH=8, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound IN-3 (105 mg, yield 92%) as a yellow oil. ).

LC-MS: m/z=235.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) 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).

Intermediate 4

(R)-1-(3-(Difluoromethyl)-2-fluoro-5-nitrophenyl)ethan-1-amine IN-4

The first step 1-bromo-3-difluoromethyl-2-fluorobenzene IN-4b

2-Fluoro-3-bromobenzaldehyde IN-4a (10.0 g, 49.26 mmol) was dissolved in dichloromethane (200 mL), cooled to 0 °C, and diethylaminosulfur trifluoride (15.9 g, 98.64 mmol), after dripping was completed, it was slowly raised to room temperature and stirred for 1 hour. TLC showed that the reaction of the starting materials was complete. The reaction solution was poured into saturated aqueous sodium bicarbonate solution and quenched, extracted with dichloromethane, 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 title compound IN-4b as a yellow liquid ( 8.1 g, yield 73%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.68 (t, J=7.2 Hz, 1H), 7.55 (t, J=7.2 Hz, 1H), 7.14 (t, J=8.0 Hz, 1H), 6.89 (t ,J=54.8Hz,1H).

The second step 1-(3-(difluoromethyl)-2-fluorophenyl)ethan-1-one IN-4c

Compound IN-4b (8.1 g, 36.0 mmol) was dissolved in 1,4-dioxane (80 mL), and triethylamine (9.1 g, 89.9 mmol) and tributyl (1-ethoxyl) were added in sequence at room temperature Ethylene) tin (15.6g, 43.2mmol), bubbled with nitrogen for 15 minutes, added bistriphenylphosphine palladium dichloride (250mg, 0.36mmol), replaced with nitrogen several times, heated to 100 ° C and stirred for 1 hour, TLC showed the reaction completely. The reaction solution was cooled to room temperature, diluted hydrochloric acid (14.4 mL, 72.0 mmol, 5 M) was added, and the mixture was stirred at room temperature for 1 hour. TLC showed that the reaction of the starting materials was complete. The reaction solution was diluted with water, 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 obtain the title compound IN-4c (5.6 g, yield 83%) as a yellow liquid. .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.00 (t, J=7.2 Hz, 1H), 7.79 (t, J=6.8 Hz, 1H), 7.34 (t, J=7.6 Hz, 1H), 6.94 (t , J=54.8Hz, 1H), 2.67(d, J=5.2Hz, 3H).

The third step 1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethan-1-one IN-4d

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

¹ H NMR (400 MHz, CDCl ₃ ) 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).

The fourth step (R,Z)-N-(1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethylene)-2-methylpropane-2-sulfinyl Amide IN-4e

Compound IN-4d (9.0 g, 38.60 mmol) was dispersed in tetraethyl titanate (90 mL), (R)-(+)-tert-butylsulfinamide (7.0 g, 57.76 mmol) was added at room temperature, and the temperature was slowly increased. After stirring to 100°C for 1 hour, TLC showed complete reaction of starting material. The reaction solution was cooled to room temperature, poured into vigorously stirred ice water, stirred continuously for 10 minutes, filtered, the filter cake was repeatedly washed with ethyl acetate, the filtrate was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Concentration gave the title compound IN-4e (9.0 g, crude) as a yellow oil, which was used directly in the next step.

LC-MS: m/z=337.1 [M+H] ⁺

The fifth step (R)-N-((R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl)-2-methylpropane-2-idene Sulfonamide IN-4f

Compound IN-4e (9.0 g, crude product) was dissolved in tetrahydrofuran (150 mL) and water (2 mL), cooled to about -60° C., and sodium borohydride (3.1 g, 81.95 mmol) was added in batches, the addition was completed, and the temperature was slowly raised to The reaction was continued for 2 hours at room temperature, TLC showed that the starting material was consumed. The reaction solution was poured into ice water and quenched, extracted with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column to obtain the title compound IN-4f (2.8 g, two step yield 21%).

LC-MS: m/z=339.1 [M+H] ⁺

The sixth step (R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethan-1-amine IN-4

Compound IN-4f (2.8 g, 8.28 mmol) was dissolved in tetrahydrofuran (30 mL), concentrated hydrochloric acid (1.4 mL, 16.52 mmol) was added, and the mixture was stirred at room temperature for 1 hour. TLC showed that the starting material was consumed. The reaction solution was cooled to room temperature, poured into saturated aqueous sodium bicarbonate solution to adjust the pH to basic, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a brown liquid, which solidified on standing to form the title compound as a brown solid IN-4 (1.8 g, 95% yield).

LC-MS: m/z=235.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) 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).

Intermediate 5

(R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethanamine IN-5

The first step (R,Z)-N-(1-(3-(difluoromethyl)-2-fluorophenyl)ethylene)-2-methylpropane-2-sulfinamide IN-5a

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

LC-MS: m/z=292.1[M+H] ⁺

The second step (R)-N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide IN-5b

Compound IN-5a (9.5g, crude product) was dissolved in tetrahydrofuran (100mL) and water (2mL), cooled to about -60°C, sodium borohydride (3.7mg, 97.80mmol) was added in batches, the addition was completed, and the temperature was slowly raised to The reaction was continued for 2 hours at room temperature, TLC showed that the starting material was consumed. The reaction solution was poured into ice water to quench, extracted with ethyl acetate, the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound IN-5b (2.9 g, two-step yield 33%).

LC-MS: m/z=294.1 [M+H] ⁺

The third step (R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethanamine IN-5

Compound IN-5b (2.9 g, 9.88 mmol) was dissolved in tetrahydrofuran (50 mL), concentrated hydrochloric acid (720 mg, 19.8 mmol) was added at room temperature, the temperature was raised to 80 °C and stirred for 1 hour, TLC showed that the starting material was consumed. The reaction solution was cooled to room temperature, poured into saturated aqueous sodium bicarbonate solution to adjust the pH to be basic, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound IN-5 (2.0 g) as a brown liquid. , the yield is 107%).

LC-MS: m/z=190.1 [M+H] ⁺

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.58 (t, J=7.4 Hz, 1H), 7.47 (t, J=6.8 Hz, 1H), 7.23 (t, J=7.6 Hz, 1H), 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).

Intermediate 6

Benzyl (S)-3-(2-ethoxy-2-oxoethoxy)pyrrolidine-1-carboxylate tert-butyl ester IN-6

The first step (S)-3-hydroxypyrrolidine-1-carboxylate benzyl ester IN-6b

(S)-1-N-tert-butoxycarbonyl-3-hydroxypyrrolidine IN-6a (5.0 g, 26.70 mmol) was dissolved in a methanolic hydrochloric acid solution (40 mL, 4 N), and the reaction was carried out at room temperature overnight. The reaction solution was concentrated, 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 at room temperature overnight. 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 obtain the title compound IN-6b (5.9 g, yield 100%) as a pale yellow oil.

The second step (S)-benzyl 3-(2-(tert-butoxy)-2-oxoethoxy)pyrrolidine-1-carboxylate IN-6c

Compound IN-6b (5.9g, 26.67mmol) was dissolved in toluene (40mL), tetrabutylammonium bromide (2.7g, 8.41mmol) and sodium hydroxide aqueous solution (24mL, 0.28mol, 12M) were added, and stirred at room temperature Tert-butyl bromoacetate (8.2 g, 42.08 mmol) was added, the addition was complete, and the mixture was stirred at room temperature overnight. 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, and concentrated. The crude product was purified by silica gel column chromatography to obtain the title compound IN-6c (10.7 g, crude product) as a pale yellow oil, which was directly used in the next step. step.

The third step (S)-2-((1-((benzyloxy)carbonyl)pyrrolidin-3-yl)oxy)acetic acid IN-6d

Compound IN-6c (10.7 g, crude product) was dissolved in dichloromethane (100 mL), trifluoroacetic acid (10 mL) was added, and the reaction was carried out at room temperature overnight. TLC showed that the reaction was basically complete. The reaction solution was concentrated (with dichloromethane 3 times) to give the title compound IN-6d (10.1 g, crude) as a pale yellow oil, which was used directly in the next step.

The fourth step (S)-3-(2-ethoxy-2-oxoethoxy) pyrrolidine-1-carboxylic acid benzyl ester IN-6

Compound IN-6d (10.1 g, crude product) was dissolved in N,N-dimethylformamide (50 mL), potassium carbonate (23.2 g, 0.17 mol) and iodoethane (11.9 g, 84.06 mmol) were added, and the reaction was carried out at room temperature. 5 hours. The reaction solution was added with water, 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 obtain the title compound IN-6 as a yellow oil (2.0 g, yield 23% for three steps) ).

Intermediate 7

(R)-Benzyl 3-(2-ethoxy-2-oxoethoxy)pyrrolidine-1-carboxylate IN-7

The first step (R)-3-hydroxypyrrolidine-1-carboxylic acid benzyl ester IN-7b

(R)-1-N-tert-butoxycarbonyl-3-hydroxypyrrolidine IN-7a (10.0 g, 53.40 mmol) was dissolved in a methanolic hydrochloric acid solution (80 mL, 4N) and reacted at room temperature overnight. The reaction solution was concentrated, 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 at room temperature overnight. 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 obtain the title compound IN-7b (9.2 g, yield 78%) as pale yellow oil.

The second step (R)-Benzyl 3-(2-(tert-butoxy)-2-oxoethoxy)pyrrolidine-1-carboxylate IN-7c

Compound IN-7b (9.2g, 41.58mmol) was dissolved in toluene (60mL), tetrabutylammonium bromide (4.0g, 12.47mmol) and sodium hydroxide aqueous solution (35mL, 0.41mol, 12M) were added, and stirred at room temperature Tert-butyl bromoacetate (12.2 g, 62.37 mmol) was added, the addition was complete, and the mixture was stirred at room temperature overnight. 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 obtain the title compound IN-7c (12.1 g, yield 86%) as pale yellow oil.

The third step (R)-2-((1-((benzyloxy)carbonyl)pyrrolidin-3-yl)oxy)acetic acid IN-7d

Compound IN-7c (6.5 g, 19.38 mmol) was dissolved in dichloromethane (70 mL), trifluoroacetic acid (7 mL) was added, and the reaction was carried out at room temperature overnight. TLC showed that the reaction was substantially complete. The reaction solution was concentrated (with dichloromethane 3 times) to give the title compound IN-7d (7.2 g, crude) as a pale yellow oil, which was used directly in the next step.

The fourth step (R)-3-(2-ethoxy-2-oxoethoxy) pyrrolidine-1-carboxylic acid benzyl ester IN-7

Compound IN-7d (7.2g, crude product) was dissolved in N,N-dimethylformamide (50mL), potassium carbonate (16.6g, 0.12mol) and iodoethane (6.0g, 38.80mmol) were added at room temperature, The reaction was carried out at room temperature for 5 hours. The reaction solution was added with water, 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 obtain the title compound IN-7 (4.2 g, 71% yield in two steps) as a yellow oil. ).

Intermediate 8

(R)-1-(3-(trifluoromethyl)phenyl)ethan-1-amine IN-8

The first step (R,Z)-2-methyl-N-(1-(3-(trifluoromethyl)phenyl)ethylene)propane-2-sulfinamide IN-8b

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 temperature was raised to 80°C and stirred for 3 hours, and the reaction was complete as detected by TLC. The reaction solution was cooled to room temperature, poured into ice water (100 mL), stirred with ethyl acetate, filtered to remove insoluble matter, the aqueous phase was extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound IN-8b (14.0 g, crude) was used directly in the next step.

The second step (R)-2-methyl-N-((R)-1-(3-(trifluoromethyl)phenyl)ethyl)propane-2-sulfinamide IN-8c

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

The third step (R)-1-(3-(trifluoromethyl)phenyl)ethan-1-amine IN-8

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

Example 1

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 1

The first step 6-chloro-N,2-dimethyl-4-pyrimidinamine 1b

4,6-Dichloro-2-methylpyrimidine 1a (20.0 g, 0.123 mol) was added to methylamine solution (100 mL, 30% ethanol solution) in batches, and reacted at room temperature for 1 hour. TLC showed that the reaction of the starting materials was completed. The reaction solution was concentrated to obtain the title compound 1b (28.0 g, crude product) as a white solid, which was directly used in the next step.

LC-MS: m/z=158.1[M+H] ⁺

The second step 6-chloro-5-iodo-N,2-methyl-4-pyrimidinamine 1c

Compound 1b (28.0 g, crude product) was dissolved in acetic acid (150 mL), NIS (33.0 g, 0.147 mol) was added in batches at room temperature, heated to 70° C. for 6 hours, and TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, adjusted to neutrality by adding saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated aqueous sodium bicarbonate solution, washed with saturated aqueous sodium sulfite solution, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain The title compound 1c (37.2 g, crude containing succinimide) as a white solid was used directly in the next step.

LC-MS: m/z=284.0[M+H] ⁺

The third step 6-chloro-N,2-dimethyl-5-vinyl-4-pyrimidinamine 1d

Compound 1c (15.0 g, crude product) was dissolved in ethylene glycol dimethyl ether (150 mL), potassium vinyl trifluoroborate (10.6 g, 79.13 mmol), potassium phosphate (28.1 g, 0.132 mol) and Pd (dppf) were added Cl ₂ dichloromethane complex (400 mg, 0.49 mmol), replaced with nitrogen for 3 times, heated to 80° C. to react overnight, TLC showed that the reaction of the starting material was completed. The reaction solution was cooled to room temperature, diluted with water, 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 silica gel column chromatography to obtain the title compound 1d as a white solid (3.87 g, yield in three steps). 40%).

LC-MS: m/z=184.1[M+H] ⁺

The fourth step 4-chloro-2-methyl-6-(methylamino)pyrimidine-5-carbaldehyde 1e

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, the solution was stirred at room temperature, and potassium osmate aqueous solution was added dropwise (catalytic amount), the reaction was performed at room temperature overnight, TLC showed that the reaction of the raw materials was completed, then sodium periodate (10.2 g, 47.69 mmol) was added, and the reaction was performed at room temperature for 1 hour, and TLC showed that the intermediate state reaction was completed. The reaction solution was added with water, 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 silica gel column chromatography to obtain the title compound 1e (3.7 g, yield 94%) as a white solid.

LC-MS: m/z=186.1[M+H] ⁺

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

Under nitrogen protection, compound 1e (900 mg, 4.85 mmol) was dissolved in tetrahydrofuran (40 mL), intermediate IN-1 (1.1 g, 6.31 mmol) was added, cooled to -60 °C, and lithium diisopropylamide (6.0 mL, 12.0 mmol, 2.0 M), after dropping, reacted at -60°C for 1 hour, TLC showed that the reaction of the starting material was completed. The reaction solution was quenched by adding citric acid, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. /1) Slurried and filtered to obtain the title compound 1f (560 mg, yield 39%) as a white solid.

LC-MS: m/z=296.1[M+H] ⁺

The sixth step 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 1g

Compound 1f (95 mg, 0.32 mmol) was dissolved in dimethyl sulfoxide (3 mL), and intermediate IN-3 (72 mg, 0.31 mmol) and N,N-diisopropylethylamine (125 mg, 0.97 mmol) were added at room temperature ), heated to 80°C and reacted for 16 hours, TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain 1 g of the title compound as a yellow solid (82 mg, yield 54%).

LC-MS: m/z=494.2[M+H] ⁺

The seventh step 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 1

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

LC-MS: m/z=464.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 7.54(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)

Example 2

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

-7(8H)-keto 2

The first step 6-chloro-2-methyl-4-pyrimidinamine 2a

4,6-Dichloro-2-methylpyrimidine 1a (15.2 g, 93.25 mmol) was added to aqueous ammonia solution (150 mL, 25%), the temperature was raised to 65° C. for 3 hours, and TLC showed that the reaction of the starting materials was completed. The reaction solution was concentrated to obtain the title compound 2a (11.9 g, crude product) as a white solid, which was directly used in the next step.

LC-MS: m/z=144.1 [M+H] ⁺

The second step 6-chloro-5-iodo-2-methyl-4-pyrimidinamine 2b

Compound 2a (11.9 g, crude product) was added to acetic acid (120 mL), N-iodosuccinimide (18.64 g, 82.85 mmol) was added in batches at room temperature, heated to 70° C. to react for 4 hours, and TLC showed that the raw materials were reacted . The reaction solution was cooled to room temperature, adjusted to neutrality by adding saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined, washed with saturated aqueous sodium bicarbonate solution, washed with aqueous sodium sulfite solution, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a white The solid title compound 2b (11.82 g, crude) was used directly in the next step.

LC-MS: m/z=270.0[M+H] ⁺

The third step 6-chloro-2-methyl-5-vinyl-4-pyrimidinamine 2c

Compound 2b (18.0 g, crude product) was dissolved in ethylene glycol dimethyl ether (180 mL), and potassium vinyl trifluoroborate (17.89 g, 0.13 mol), potassium phosphate (35.9 g, 0.17 mol) and Pd were sequentially added at room temperature (dppf) Cl ₂ dichloromethane complex (1.64 g, 2.01 mmol), replaced with nitrogen three times, heated to 80° C. for 3 hours, and TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 2c (6.1 g, yield in three steps) as a white solid. 25%).

The fourth step 4-amino-6-chloro-2-methylpyrimidine-5-carbaldehyde 2d

Compound 2c (5.00g, 29.48mmol) was dissolved in acetone and water (120mL/30mL), sodium periodate (14.32g, 66.95mmol) and potassium osmate aqueous solution (catalytic amount) were added, and the reaction was carried out at room temperature for 2 hours. TLC showed The raw materials reacted completely. The reaction solution was added with water, 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 silica gel column chromatography to obtain the title compound 2d (2.5 g, yield 49%) as an off-white solid.

LC-MS: m/z=172.1[M+H] ⁺

The fifth step (S)-4-chloro-2-methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 2e

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, cooled to -60 °C, and diisopropylamino was added dropwise. Lithium (22 mL, 43.86 mmol, 2 M) was dropped at -60°C for 1 hour, and then slowly warmed to room temperature for overnight reaction. The reaction solution was quenched by adding citric acid, 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 obtain the title compound 2e as a pale yellow solid (1.0 g, yield twenty four%).

LC-MS: m/z=282.1[M+H] ⁺

The sixth step 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

Compound 2e (100 mg, 0.35 mmol) was dissolved in dimethyl sulfoxide (3 mL), and intermediate IN-3 (83 mg, 0.35 mmol) and N,N-diisopropylethylamine (160 mg, 1.24 mmol) were added at room temperature ), 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, 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 obtain the title compound 2f (80 mg, yield 48%) as a yellow solid.

LC-MS: m/z=480.1[M+H] ⁺

The seventh step 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

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

LC-MS: m/z=450.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 7.57(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)

Example 3

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

The first step 4-chloro-6-((4-methoxybenzyl)amino)pyrimidine-5-carbaldehyde 3b

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

LC-MS: m/z=278.1[M+H] ⁺

Second Step (S)-4-Chloro-8-(4-methoxybenzyl)-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidine-7(8H )-keto 3c

Under nitrogen protection, compound 3b (2.00 g, 7.20 mmol) was dissolved in tetrahydrofuran (300 mL), intermediate IN-1 (1.63 g, 9.36 mmol) was added at room temperature, cooled to -10 °C, and diisopropylamino was added dropwise. Lithium (10.8 mL, 21.60 mmol, 2M) was dropped, reacted at -10 °C for 3 hours, and then slowly returned to room temperature to react overnight. The reaction solution was quenched by adding citric acid, 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 obtain the title compound 3c (900 mg, yield 32%) as a brown-yellow solid. ).

LC-MS: m/z=388.1[M+H] ⁺

The third step 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

Compound 3c (500 mg, 1.29 mmol) was dissolved in dimethyl sulfoxide (6 mL), and intermediate IN-3 (300 mg, 1.28 mmol) and N,N-diisopropylethylamine (579 mg, 4.48 mmol) were added in sequence at room temperature mmol), 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, 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 obtain the title compound 3d (580 mg, yield 77%) as a yellow solid. .

LC-MS: m/z=586.2[M+H] ⁺

The fourth step 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

Compound 3d (580 mg, 0.99 mmol) was dissolved in trifluoroacetic acid (10 mL), heated to 75° C. and reacted for 2 hours. TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, and saturated aqueous sodium bicarbonate solution was added dropwise to adjust the pH to basic, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound 3e (430 mg, crude product) as a gray-brown solid. ), used directly in the next step.

LCMS: m/z=466.2[M+H] ⁺

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

Compound 3e (80 mg, crude product) 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 reaction was heated to 90 °C for 3 hours , TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, water was added, 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 3 (42 mg, two-step yield 52) %).

LC-MS: m/z=436.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.20 (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)

Example 4

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 4

The first step 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

Compound 3e (100 mg, 0.21 mmol) was dissolved in N,N-dimethylformamide (3 mL), potassium carbonate (89 mg, 0.64 mmol) and methyl iodide (61 mg, 0.43 mmol) were added, and the reaction was carried out at room temperature overnight. TLC showed the starting material. The reaction is complete. The reaction solution was added with water, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 4a (91 mg, crude product) as a yellow solid, which was used directly in the next step.

LC-MS: m/z=480.2[M+H] ⁺

The second step 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 4

Compound 4a (91 mg, crude product) 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 reaction was heated to 90 °C for 3 hours. , TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 4 (42 mg, two-step yield 45) %).

LC-MS: m/z=450.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.31-8.25(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)

Example 5

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

The first step 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

Compound 3c (120 mg, 0.31 mmol) was dissolved in dimethyl sulfoxide (3 mL), and intermediate IN-4 (73 mg, 0.31 mmol) and N,N-diisopropylethylamine (140 mg, 1.08 mmol) were added at room temperature ), heated to 100°C and reacted for 8 hours, TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 5a (66 mg, yield 36%) as a yellow solid.

LC-MS: m/z=586.3[M+H] ⁺

The second step 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

Compound 5a (66 mg, 0.11 mmol) was dissolved in trifluoroacetic acid (2 mL), heated to 75° C. and reacted for 2 hours. TLC showed that the reaction of the starting material was complete. The reaction solution was cooled to room temperature, and saturated aqueous sodium bicarbonate solution was added dropwise to adjust the pH to basic, 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 obtain a gray-brown solid The title compound 5b (27 mg, 52% yield).

LC-MS: m/z=466.2[M+H] ⁺

The third step 4-(((R)-1-(3-(difluoromethyl)-2-fluoro-5-nitrophenyl)ethyl)amino)-8-methyl-6-(((( S)-Tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 5c

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

LC-MS: m/z=480.2[M+H] ⁺

The fourth step 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

Compound 5c (30 mg, crude product) 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 reaction was heated to 90 °C for 3 hours. , TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 5 (12 mg, two-step yield: 46 %).

LC-MS: m/z=450.3 [M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.26(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), 5.19-5.13(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 = 6.8 Hz, 3H). (96.10% purity by HPLC)

Example 6

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

The first step (E)-methyl 3-(4-chloro-2-methyl-6-(methylamino)pyrimidin-5-yl)acrylate 6a

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, and nitrogen was replaced. Three times, methyl acrylate (3.95 g, 45.88 mmol) was added with a syringe, heated to 100° C. to react overnight, and TLC showed that the reaction of the raw materials was complete. The reaction solution was cooled to room temperature, water was added, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, the crude product was slurried with a mixed solution (petroleum ether/ethyl acetate=5/1), filtered, filtered The cake was washed and dried to give the title compound 6a (6.0 g, crude) as a khaki solid, which was used directly in the next step.

LC-MS: m/z=242.1[M+H] ⁺

The second step 4-methoxy-2,8-dimethylpyrido[2,3-d]pyrimidin-7(8H)-one 6b

Compound 6a (2.17 g, crude product) was added to methanol (30 mL), cooled to 0 °C, sodium hydride (754 mg, 18.85 mmol, 60%) was added in batches, the addition was completed, heated to 60 °C and reacted for 6 hours, LCMS showed that the raw materials reacted completely. The reaction solution was cooled to 0°C, quenched by dropwise addition of water, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 6b (1.80 g, crude product) as a pale yellow solid, which was directly used in the next step. step.

LC-MS: m/z=206.1[M+H] ⁺

The third step 6-bromo-4-methoxy-2,8-dimethylpyrido[2,3-d]pyrimidin-7(8H)-one 6c

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

The fourth step 6-bromo-4-hydroxy-2,8-dimethylpyrido[2,3-d]pyrimidin-7(8H)-one 6d

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

LC-MS: m/z=272.0[M+H] ⁺

The fifth step 6-bromo-2,8-dimethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-4-yl 2,4,6-triisopropyl besylate 6e

Compound 6d (1.78 g, crude product) 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, cooled to 0 °C, 2,4,6-Triisopropylbenzenesulfonyl chloride (2.46 g, 8.12 mmol) was added in batches, the addition was completed, the reaction was raised to room temperature for 0.5 hours, and TLC showed that the reaction of the raw materials was completed. The reaction solution was added with water, extracted with dichloromethane, 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 title compound 6e (2.45 g, five-step yield 59%) as a white solid.

LC-MS: m/z=536.0[M+H] ⁺

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

Compound 6e (1.45g, 2.70mmol) was dissolved in dimethyl sulfoxide (20mL), and intermediate IN-3 (600mg, 2.56mmol) and N,N-diisopropylethylamine (1.15g, 2.56mmol) were added at room temperature 8.90 mmol), heated to 80°C and reacted for 3 hours, TLC showed that the reaction of the starting material was completed. The reaction solution was cooled to room temperature, water was added, 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 silica gel column chromatography to obtain the title compound 6f (1.08 g, yield 86%) as a pale yellow solid ).

LC-MS: m/z=486.0[M+H] ⁺

The seventh step (R)-5-(2,8-dimethyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7-oxygen Substituted-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate tert-butyl ester 6h

Under nitrogen protection, compound 6f (200mg, 0.41mmol) and 1-tert-butoxycarbonyl-3,6-dihydro-2H-pyridine-5-boronic acid pinacol ester 6g (190mg, 0.61mmol) were dissolved in 1, In 4-dioxane (15 mL) and water (3 mL), Pd(dppf)Cl ₂ dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (130 mg, 1.23 mmol) were added at room temperature, and the temperature was raised to 100 After stirring at °C for 1 hour, the reaction was complete as detected by TLC. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 6h (251 mg, crude product) as a yellow solid, which was used directly with in the next step.

LC-MS: m/z=589.3[M+H] ⁺

The eighth step 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

Compound 6h (251 mg, crude product) was dissolved in ethyl acetate (10 mL), palladium/carbon (100 mg, 10%) was added at room temperature, the temperature was raised to 50 °C under a hydrogen atmosphere, and the reaction was performed overnight. LCMS detected that the reaction was complete. The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with ethyl acetate, and the filtrate was concentrated to obtain the title compound 6i (260 mg, crude product) as a pale yellow oil, which was directly used in the next step.

LC-MS: m/z=561.3[M+H] ⁺

The ninth step 3-(4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-7-oxygen Substituted-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylate tert-butyl ester 6j

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

LC-MS: m/z=603.3[M+H] ⁺

The tenth step 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

Compound 6j (270 mg, crude product) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for 2 hours. TLC showed that the reaction of the starting material was completed. The reaction solution was adjusted to basicity by adding saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 6k (200 mg, crude product) as a yellow solid, which was directly used in the next step. .

The eleventh step 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 6l

Compound 6k (200 mg, crude product) was dissolved in methanol (5 mL), aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium/carbon (70 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC showed the reaction of the starting materials. Finish. The reaction solution was filtered, and the filtrate was concentrated to obtain the title compound 61 (190 mg, crude product) as a yellow oil, which was directly used in the next step.

LC-MS: m/z=517.3 [M+H] ⁺

The twelfth step 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-dimethyl-6-(1-methyl) ylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 6

Compound 61 (190 mg, crude product) was dissolved in methanol (6 mL), sodium hydroxide aqueous solution (2 mL, 8.0 mmol, 4 N) was added at room temperature, heated to 80 °C and reacted overnight, TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 6 (85 mg, yield 44% for six steps) ).

LC-MS: m/z=475.3 [M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.10(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 by HPLC)

Example 7

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

The first step 2-fluoro-5-nitro-3-(trifluoromethyl)benzaldehyde 7b

At 0°C, 2-fluoro-3-trifluoromethylbenzaldehyde 7a (3 g, 15.62 mmol) was dissolved in concentrated sulfuric acid (6 mL), and nitric acid (2.3 mL, 68%) was added dropwise. The reaction was carried out at room temperature for 5 hours, and TLC monitoring showed a small amount of starting material remaining. The reaction solution was slowly added with ice water, 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 obtain the title compound 7b (2.2 g, yield 59%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 10.42 (s, 1H), 8.96-8.94 (m, 1H), 8.77-8.75 (m, 1H).

The second step (2-fluoro-5-nitro-3-(trifluoromethyl)phenyl)methanol 7c

Compound 7b (940 mg, 3.96 mmol) was dissolved in absolute ethanol (15 mL), cooled to 0 °C, sodium borohydride (226 mg, 5.97 mmol) was added in batches, the addition was completed, and the reaction was continued at 0 °C for 35 minutes, TLC showed the raw material Conversion is complete. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride solution, concentrated to remove ethanol, the aqueous phase 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 obtain the title of yellow liquid Compound 7c (792 mg, 84% yield).

The third step (5-amino-2-fluoro-3-(trifluoromethyl)phenyl)methanol 7d

Compound 7c (792 mg, 3.31 mmol) was dissolved in absolute ethanol (18 mL), palladium/carbon (400 mg, 10%) was added, and the reaction was carried out overnight at room temperature under a hydrogen atmosphere. TLC showed complete conversion of the starting material. The reaction solution was filtered with celite, the filter cake was washed with ethanol, and the filtrate was concentrated to obtain the title compound 7d (656 mg, crude product), which was directly used in the next step.

LC-MS: m/z=210.1[M+H] ⁺

The fourth step (4-fluoro-3-(hydroxymethyl)-5-(trifluoromethyl)phenyl) benzyl carbamate 7e

Compound 7d (731 mg, crude product), potassium carbonate (724 mg, 5.24 mmol) and benzyl chloroformate (716 mg, 4.20 mmol) were sequentially added to tetrahydrofuran (20 mL) and reacted at room temperature for 3 hours. TLC showed that the reaction was complete. The reaction solution was added with water, 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 obtain the title compound 7e (1.06 g, two-step yield 84%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ 7.64(t, J=5.6Hz, 2H), 7.40-7.33(m, 5H), 6.90(s, 1H), 5.20(s, 2H), 4.77(s, 2H).

The fifth step (4-fluoro-3-formyl-5-(trifluoromethyl) phenyl) benzyl carbamate 7f

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

The sixth step (4-fluoro-3-(1-hydroxyethyl)-5-(trifluoromethyl)phenyl) benzyl carbamate 7g

Compound 7f (825 mg, 2.42 mmol) was dissolved in anhydrous tetrahydrofuran (18 mL), cooled to 0 °C, methylmagnesium bromide (2.5 mL, 7.5 mmol, 3M tetrahydrofuran solution) was added dropwise, the drop was completed, and the reaction was raised to room temperature overnight. , TLC showed that the conversion of starting material was complete. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride solution, 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 obtain the title compound 7 g (858 mg, yield) as a white solid rate 99%).

¹ H NMR (400MHz, CDCl ₃ ) δ 7.67-7.65 (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).

The seventh step (3-acetyl-4-fluoro-5-(trifluoromethyl)phenyl) benzyl carbamate 7h

Compound 7g (858 mg, 2.40 mmol) was dissolved in chloroform (20 mL), manganese dioxide (3.2 g, 36.78 mmol) was added, and the reaction was carried out at room temperature overnight. TLC showed that the conversion of the raw material was not complete. After heating under reflux for 6 hours, TLC showed that the conversion of the raw material was complete. The reaction solution was cooled to room temperature, and a pad of celite was used for filtration, the filter cake was washed with dichloromethane, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 7h (680 mg, yield 80%).

¹ H NMR (400MHz, CDCl ₃ ) δ 8.17(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).

The eighth step (R,E)-(3-(1-((tert-butylsulfinyl)imino)ethyl)-4-fluoro-5-(trifluoromethyl)phenyl)carbamic acid benzyl ester 7i

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 reaction was heated to 60°C overnight, TLC showed complete conversion of starting material. The reaction solution was cooled to room temperature, quenched by dropwise addition of water, 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 obtain the title compound 7i (343 mg, yield 43%). ).

The ninth step (3-((R)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-4-fluoro-5-(trifluoromethyl)phenyl)benzylcarbamate Ester 7j

Compound 7i (343 mg, 0.75 mmol) was dissolved in tetrahydrofuran (3 mL) and water (2 drops), cooled to -70°C, sodium borohydride (114 mg, 3.01 mmol) was added in portions, and the reaction was continued for 1 hour. TLC showed that the conversion of the starting material was not completed, so sodium borohydride (114 mg, 3.01 mmol) was added, and the temperature was gradually raised to -45 °C for 1 hour. TLC showed that the starting material had been converted, and two new spots were formed (LCMS confirmed that they were two isomers). , the ratio is about 3:1, and the desired configuration accounts for the main ratio). The reaction was quenched by dropwise addition of saturated ammonium chloride solution, 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 obtain the title compound 7j (122 mg, yield 35 %).

LC-MS: m/z=461.1[M+H] ⁺

The tenth step (R)-(3-(1-aminoethyl)-4-fluoro-5-(trifluoromethyl)phenyl)carbamic acid benzyl ester 7k

Compound 7j (122 mg, 0.26 mmol) was dissolved in tetrahydrofuran (1 mL), concentrated hydrochloric acid (0.3 mL) was added dropwise at room temperature, and the reaction was continued at room temperature for 40 minutes. TLC showed that the conversion of the starting materials was completed. The reaction was quenched by dropwise addition of saturated sodium carbonate solution, 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 obtain the title compound 7k (92 mg, yield 97%). ).

The eleventh step (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 7l

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, and the temperature was raised to 100 ° C for 5 hours. TLC It shows that the reaction of the starting materials is complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 71 (48 mg, yield 28%) as a white solid.

The twelfth step 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

Compound 71 (48 mg, 0.078 mmol) was dissolved in methanol (5 mL), palladium/carbon (60 mg, 10%) was added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction solution was filtered through a pad of celite, the filtrate was concentrated, and the crude product was purified by Prep-TLC to give the title compound 7 (14 mg, yield 37%) as a white solid.

LC-MS: m/z=482.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 7.60 (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), 4.07-3.98(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=7.2 Hz, 3H). (94.15% purity by HPLC)

Example 8

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)pyrrolidin-3-yl)acetamide 8

The first step ((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)pyrrolidin-3-yl)carbamate tert-butyl ester 8b

Compound 6f (300 mg, 0.62 mmol) was dissolved in toluene (30 mL), (R)-3-tert-butoxycarbonylaminopyrrolidine 8a (172 mg, 0.92 mmol) was added at room temperature, Pd ₂ (dba) ₃ (56 mg, 0.061 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (57 mg, 0.092 mmol) and sodium tert-butoxide (178 mg, 1.85 mmol), replaced with nitrogen three times, heated to 100 ° C for reaction overnight, TLC showed complete reaction of starting material. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 8b (185 mg, yield 51%) as a brown solid. .

LC-MS: m/z=592.3[M+H] ⁺

The second step 6-((R)-3-aminopyrrolidin-1-yl)-2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoro) Methyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 8c

Compound 8b (172 mg, 0.29 mmol) was dissolved in a methanol solution of hydrochloric acid (3 mL, 4 M) and reacted at room temperature for 1 hour. TLC showed that the reaction of the starting material was complete. The reaction was adjusted to basicity by adding saturated aqueous sodium bicarbonate solution dropwise, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound 8c (160 mg, crude product) as a brown solid, which was used directly in the next step. step.

LC-MS: m/z=492.2[M+H] ⁺

The third step N-((R)-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)pyrrolidin-3-yl)acetamide 8d

Compound 8c (160 mg, crude product) was dissolved in dichloromethane (5 mL), triethylamine (98 mg, 0.97 mmol) and acetic anhydride (50 mg, 0.49 mmol) were added, and the reaction was carried out at room temperature for 1 hour. TLC showed that the reaction of the starting materials was completed. The reaction solution was diluted with dichloromethane, washed with hydrochloric acid (1N), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, the crude product was slurried with a mixed solution (dichloromethane/petroleum ether=1/10), filtered, and the filter cake Washing and drying gave the title compound 8d (120 mg, crude) as a yellow solid, which was used directly in the next step.

LC-MS: m/z=534.2[M+H] ⁺

The fourth step N-((R)-1-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-di Methyl-7-oxo7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidin-3-yl)acetamide 8

Compound 8d (120 mg, crude product) was dissolved in ethyl acetate (3 mL) and methanol (1 mL), concentrated hydrochloric acid (3 drops) and palladium/carbon (80 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC showed The reaction of the raw materials is completed. The reaction solution was filtered through a pad of celite, and the filtrate was adjusted to alkaline by adding ammonia methanol solution (1 mL) dropwise, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 8 (50 mg, three-step yield 34%) as a white solid.

LC-MS: m/z=504.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 6.99(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)

Example 9

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)pyrrolidin-3-yl)acetamide 9

The first step ((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)pyrrolidin-3-yl)carbamate tert-butyl ester 9b

Compound 6f (300 mg, 0.62 mmol) was dissolved in toluene (30 mL), (S)-3-tert-butoxycarbonylaminopyrrolidine 9a (172 mg, 0.92 mmol), Pd ₂ (dba) ₃ (56 mg, 0.061 mmol) were added , 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (57 mg, 0.092 mmol) and sodium tert-butoxide (178 mg, 1.85 mmol), replaced with nitrogen three times, heated to 100 °C for overnight reaction, TLC showed The reaction of the raw materials is completed. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 9b (105 mg, yield 29%) as a brown solid. .

LC-MS: m/z=592.3[M+H] ⁺

The second step 6-((S)-3-aminopyrrolidin-1-yl)-2,8-dimethyl-4-(((R)-1-(3-nitro-5-(trifluoro) Methyl)phenyl)ethyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one 9c

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

The third step N-((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)pyrrolidin-3-yl)acetamide 9d

Compound 9c (90 mg, crude product) was dissolved in dichloromethane (3 mL), triethylamine (98 mg, 0.97 mmol) and acetic anhydride (50 mg, 0.49 mmol) were added, and the reaction was carried out at room temperature for 1 hour. TLC showed that the reaction of the starting materials was completed. The reaction solution was diluted with dichloromethane, washed with 1N hydrochloric acid, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated.

LC-MS: m/z=534.2[M+H] ⁺

The fourth step N-((S)-1-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2,8-di Methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyrrolidin-3-yl)acetamide 9

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

LC-MS: m/z=504.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 6.99(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)

Example 10

(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

The first step (E)-methyl 3-(4-chloro-6-((4-methoxybenzyl)amino)pyrimidin-5-yl)acrylate 10a

Compound 3b (7.41 g, 26.68 mmol) was dissolved in tetrahydrofuran (100 mL), methoxyformylmethylene triphenylphosphine (11.60 g, 34.69 mol) was added at room temperature, heated to 65° C. to react for 2 hours, TLC showed the starting material The reaction is complete. The reaction solution was cooled to room temperature, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 10a (8.61 g, yield 97%) as a pale yellow oil.

LC-MS: m/z=334.1[M+H] ⁺

The second step 4-methoxy-8-(4-methoxybenzyl)pyrido[2,3-d]pyrimidin-7(8H)-one 10b

Compound 10a (8.61 g, 25.80 mmol) was dissolved in methanol (100 mL), cooled to 0 °C, sodium hydride (2.27 g, 56.75 mmol, 60%) was added, the temperature was raised to 65 °C and the reaction was performed overnight, TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, quenched by adding water, stirred for 0.5 hours, filtered, and the filter cake was washed with water, dissolved in dichloromethane/methanol, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 10b (7.67 g, crude product) as a white solid, which was directly for the next step.

The third step 4-methoxypyrido[2,3-d]pyrimidin-7(8H)-one 10c

Compound 10b (7.67 g, crude product) was dissolved in trifluoroacetic acid (60 mL), and the temperature was raised to 75° C. and reacted for 3 hours. TLC showed that the reaction of the starting materials was completed. The reaction solution was concentrated, neutralized by adding saturated aqueous sodium bicarbonate solution, adding dichloromethane (the product was insoluble in dichloromethane), stirring for 30 minutes, filtering, washing the filter cake with water, washing with dichloromethane, and drying to obtain the title compound 10c as a white solid ( 3.90g, crude product), used directly in the next step.

LC-MS: m/z=178.1[M+H] ⁺

The fourth step 4-methoxy-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one 10d

Compound 10c (3.90 g, crude product) was dissolved in N,N-dimethylformamide (40 mL), potassium carbonate (4.56 g, 32.99 mmol) and methyl iodide (6.25 g, 44.03 mmol) were added at room temperature, and heated to 50 The reaction was carried out at °C for 1 hour, and TLC showed that the reaction of the starting material was complete. The reaction solution was cooled to room temperature, water was added, a solid was precipitated, filtered, and the filter cake was washed with water, then dissolved in dichloromethane, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 10d (3.04 g, crude product) as a white solid, which was directly used in the next step. step.

The fifth step 6-bromo-4-hydroxy-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one 10e

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

The sixth step 6-bromo-4-chloro-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one 10f

Compound 10e (2.07 g, crude product) was suspended in 1,4-dioxane (60 mL), and N,N-diisopropylethylamine (4.17 g, 32.27 mmol) and phosphorus oxychloride (4.95 mmol) were added at room temperature g, 32.28 mmol), heated to 80° C. to react for 4 hours, TLC showed that the reaction of the starting material was completed. The reaction solution was cooled, and saturated aqueous sodium bicarbonate solution was added dropwise at low temperature to make it basic, 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 obtain the title as a pale yellow solid. Compound 10f (1.5 g, 26% yield over five steps).

LC-MS: m/z=274.0[M+H] ⁺

¹ H NMR (400MHz, CDCl ₃ ) δ 8.87(s,1H), 8.46(s,1H), 3.87(s,3H).

The seventh step (R)-6-bromo-8-methyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[2,3 -d]Pyrimidine-7(8H)-one 10g

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

LCMS: m/z=472.1[M+H] ⁺

The eighth step (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

Compound 10g (150mg, 0.32mmol) and 4-methylcyclohex-1-enylboronic acid pinacol ester 10h (106mg, 0.48mmol) were dissolved in 1,4-dioxane (10mL) and water (2mL) , Pd(dppf)Cl ₂ dichloromethane complex (18 mg, 0.022 mmol) and sodium carbonate (148 mg, 1.40 mmol) were added at room temperature, and the temperature was raised to 100° C. to react for 1 hour. TLC detected that the reaction was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 10i (120 mg, yield 77%) as a brown oil. ).

LC-MS: m/z=489.2[M+H] ⁺

The ninth step (R)-4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpiperidine- 4-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 14

Compound 10i (120 mg, 0.25 mmol) was dissolved in methanol (5 mL), palladium/carbon (50 mg, 10%) was added, and the temperature was raised to 30° C. for 2 hours under a hydrogen atmosphere. The reaction was completed by LCMS. The reaction solution was filtered through celite, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain a crude yellow solid, which was dissolved with dilute hydrochloric acid (1N), extracted with ethyl acetate, discarded the organic phase, and the aqueous phase was saturated with sodium bicarbonate The aqueous solution was adjusted to basic, 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 14 (31 mg, yield 27%) as a pale yellow solid.

LC-MS: m/z=461.3[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.33(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)

Example 11

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

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

Compound 10g (200mg, 0.42mmol) and 1-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-boronic acid pinacol ester 11a (187mg, 0.63mmol) were dissolved in 1,4-dioxane To the ring (10 mL) and water (2 mL), add Pd(dppf)Cl ₂ dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (130 mg, 1.23 mmol) at room temperature, replace with nitrogen three times, and heat up to 100 The reaction was carried out at °C for 1 hour, and the reaction was complete as detected by TLC. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 11b (200 mg, yield 84%) as a yellow solid. .

LC-MS: m/z=561.3[M+H] ⁺

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

Compound 11b (200 mg, 0.36 mmol) was dissolved in ethyl acetate (10 mL), palladium/carbon (50 mg, 10%) was added at room temperature, and the temperature was raised to 50 °C under a hydrogen atmosphere to react overnight. TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with ethyl acetate, and the filtrate was concentrated to obtain the title compound 11c (170 mg, crude product) as a yellow solid, which was directly used in the next step.

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

The third step (R)-3-(4-((1-(3-acetylamino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7 ,8-Dihydropyrido[2,3-d]pyrimidin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester 11d

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

LC-MS: m/z=573.3 [M+H] ⁺

The fourth step 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-carboxylate tert-butyl ester 11e

Compound 11d (207 mg, crude product) was dissolved in methanol (10 mL), palladium/carbon (50 mg, 10%) was added, the temperature was raised to 30 °C under a hydrogen atmosphere, and the reaction was performed for 2 hours. TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain the title compound 11e (200 mg, crude product) as a yellow solid, which was directly used in the next step.

The fifth step N-(3-((1R)-1-((8-methyl-7-oxo-6-(pyrrolidin-3-yl)-7,8-dihydropyrido[2,3 -d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 11f

Compound 11e (200 mg, crude product) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for 2 hours. TLC showed that the reaction of the starting materials was completed. The reaction solution was adjusted to basicity by adding saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 11f (187 mg, crude product) as a yellow solid, which was directly used in the next step .

The sixth step N-(3-((1R)-1-((8-methyl-6-(1-methylpyrrolidin-3-yl)-7-oxo-7,8-dihydropyrido [2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 11g

Compound 11f (187 mg, crude product) was dissolved in methanol (5 mL), aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium/carbon (70 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC showed the reaction of the starting materials. Finish. The reaction solution was filtered through a pad of celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain 11 g (190 mg, crude product) of the title compound as a yellow oil, which was directly used in the next step.

The seventh step 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-6-(1-methylpyrrolidine- 3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 11

Compound 11g (190mg, crude product) was dissolved in methanol (6mL), sodium hydroxide aqueous solution (3mL, 4N) was added at room temperature, heated to 70°C and reacted overnight, TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 11 (60 mg, yield 40% for six steps) .

LC-MS: m/z=447.3[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.34(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), 2.52-2.40(m, 1H), 2.26-2.15(m, 1H), 1.62(d, J=7.2Hz, 3H).(94.63%purity by HPLC)

Example 12

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

The compound of Example 12 was synthesized with reference to the synthesis method of Example 13.

Example 13

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

The first step 4-chloro-6-(methylamino)pyrimidine-5-carbaldehyde 13a

4,6-Dichloro-5-pyrimidinecarboxaldehyde 3a (52.0g, 0.29mol) was dissolved in dichloromethane (500mL), cooled to 0°C, triethylamine (59.5g, 0.59mol) was added dropwise, and added in batches Methylamine hydrochloride (16.9 g, 0.25 mol) was added and reacted at 0°C for 2 hours. TLC showed that the reaction was complete. The reaction solution was filtered through a pad of celite, the filtrate was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography to obtain a crude product. Washing with butyl ether gave the title compound 13a (23.5 g, 46% yield) as a pale yellow solid.

LCMS: m/z=172.1[M+H] ⁺

Second Step (S)-3-((4-Chloro-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrole Benzyl alkane-1-carboxylate 13b

Under nitrogen protection, compound 13a (900 mg, 5.24 mmol) was dissolved in tetrahydrofuran (15 mL), intermediate IN-6 (2.1 g, 6.81 mmol) was added, cooled to -15 °C, and lithium diisopropylamide (6.6 mmol) was added dropwise. mL, 13.20 mmol, 2.0 M), after dropping, reacted at -10 °C for 5 hours, and reacted at room temperature overnight, TLC showed that the reaction was complete. The reaction was quenched by dropwise addition of saturated aqueous citric acid solution, 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 obtain the title compound 13b (144 mg, yield) as a yellow oil. 7%).

LCMS: m/z=415.1[M+H] ⁺

The third step (S)-3-((8-methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7 -oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylate benzyl ester 13c

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

The fourth step (S)-3-((4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7- Benzyl oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylate 13d

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 amine chloride (86 mg, 1.58 mmol) were added at room temperature, and the temperature was raised to 90 ° C for reaction 5 After hours, TLC showed the reaction was complete. The reaction solution was cooled to room temperature, filtered through a pad of celite, water was added to the filtrate, 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 13d as a brown solid. (140 mg, 76% yield).

The fifth step (S)-3-((4-(((R)-1-(3-acetylamino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7 -oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylate benzyl ester 13e

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

The sixth step N-(3-((R)-1-((8-methyl-7-oxo-6-(((S)-pyrrolidin-3-yl)oxy)-7,8- Dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 13f

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

The seventh step N-(3-((R)-1-((8-methyl-6-(((S)-1-methylpyrrolidin-3-yl)oxy)-7-oxo- 7,8-Dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 13g

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

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

Compound 13g (95mg, crude product) was dissolved in methanol (4mL), sodium hydroxide aqueous solution (4ml, 16.00mmol, 4N) was added at room temperature, the temperature was raised to 80°C and the reaction was performed overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 13 (40 mg, three-step yield 52) %).

LCMS: m/z=463.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.29(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).

Example 14

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

The first step (R)-3-((4-chloro-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrole Benzyl alkane-1-carboxylate 14a

Under nitrogen protection, compound 13a (700 mg, 4.08 mmol) was dissolved in tetrahydrofuran (40 mL), intermediate IN-7 (1.6 g, 5.30 mmol) was added, cooled to -15 °C, and lithium diisopropylamide (5.1 mmol) was added dropwise. mL, 10.20 mmol, 2.0 M), after dropping, reacted at -10 °C for 5 hours, and reacted at room temperature overnight, TLC showed that the reaction was complete. The reaction was quenched by dropwise addition of saturated aqueous citric acid solution, 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 obtain the title compound 14a (176 mg, yield) as a yellow oil. 10%).

LCMS: m/z=415.2[M+H] ⁺

The second step (R)-3-((8-methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-7 -oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylate benzyl ester 14b

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

LCMS: m/z=613.2[M+H] ⁺

The third step (R)-3-((4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7- Benzyl oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylate 14c

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 amine chloride (111 mg, 2.06 mmol) were added at room temperature, and the temperature was raised to 90 ° C for reaction 5 After hours, TLC showed the reaction was complete. The reaction solution was cooled to room temperature, filtered through a pad of celite, water was added to the filtrate, 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 obtain the title compound 14c as a brown solid. (125 mg, 52% yield).

LCMS: m/z=583.3[M+H] ⁺

The fourth step (R)-3-((4-(((R)-1-(3-acetylamino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7 -oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)oxy)pyrrolidine-1-carboxylate benzyl ester 14d

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

The fifth step N-(3-((R)-1-((8-methyl-7-oxo-6-(((R)-pyrrolidin-3-yl)oxy)-7,8- Dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 14e

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

The sixth step N-(3-((R)-1-((8-methyl-6-(((R)-1-methylpyrrolidin-3-yl)oxy)-7-oxo- 7,8-Dihydropyrido[2,3-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 14f

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

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

Compound 14f (90 mg, crude product) was dissolved in methanol (4 mL), sodium hydroxide aqueous solution (4 mL, 16.00 mmol, 4 N) was added, the temperature was raised to 80° C. and reacted overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 14 (35 mg, three-step yield 59 %).

LCMS: m/z=463.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.29(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).

Example 15

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

The first step (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)-carboxylate tert-butyl ester 15a

Compound 10g (240mg, 0.51mmol) and 1-tert-butoxycarbonyl-3,6-dihydro-2H-pyridine-5-boronic acid pinacol ester 6g (236mg, 0.76mmol) dissolved in 1,4-dioxane To the ring (10 mL) and water (2 mL), add Pd(dppf)Cl ₂ dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (135 mg, 1.27 mmol) at room temperature, replace with nitrogen three times, and heat up to 100 The reaction was carried out at °C for 1 hour, and the reaction was complete as detected by TLC. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 15a (260 mg, yield 89%) as a yellow solid. .

LC-MS: m/z=575.3 [M+H] ⁺

The second step (R)-5-(4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7, 8-Dihydropyrido[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate tert-butyl ester 15b

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 amine chloride (121 mg, 2.26 mmol) were added at room temperature, and the temperature was raised to 90 ° C for reaction 4 After hours, TLC showed that the starting material had reacted. The reaction solution was cooled to room temperature, filtered through a pad of celite, water was added to the filtrate, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound 15b (253 mg, crude product) as a yellow solid, which was directly used for Next step.

LC-MS: m/z=545.3 [M+H] ⁺

The third step (R)-5-(4-((1-(3-acetylamino-5-(trifluoromethyl)phenyl)ethyl)amino)-8-methyl-7-oxo-7 ,8-Dihydropyrido[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate tert-butyl ester 15c

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

The fourth step 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-carboxylate tert-butyl ester 15d

Compound 15c (250 mg, crude product) was dissolved in methanol (10 mL), palladium/carbon (80 mg, 10%) was added at room temperature, and the temperature was raised to 30 °C under a hydrogen atmosphere for 2 hours. TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain the title compound 15d (240 mg, crude product) as a yellow solid, which was directly used in the next step.

The fifth step 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

Compound 15d (240 mg, crude product) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for 2 hours. TLC showed that the reaction of the starting materials was completed. The reaction solution was adjusted to basicity by adding saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 15e (180 mg, crude product) as a yellow solid, which was directly used in the next step.

LC-MS: m/z=489.3[M+H] ⁺

The sixth step 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

Compound 15e (180 mg, crude product) was dissolved in methanol (5 mL), aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium/carbon (70 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC showed the reaction of the starting materials. Finish. The reaction solution was filtered through a pad of celite, the filter cake was washed with methanol, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 15f (142 mg, five-step yield 63%) as a pale yellow solid.

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

Compound 15f (142 mg, 0.28 mmol) was dissolved in methanol (6 mL), sodium hydroxide aqueous solution (3 mL, 12 mmol, 4 N) was added at room temperature, heated to 70 °C and reacted overnight, TLC showed that the reaction of the starting materials was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 15 (80 mg, yield 62%) as a white solid.

LC-MS: m/z=461.3[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.33(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)

Example 16

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

The first step (R)-6-bromo-4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methylpyrido[2,3- d]Pyrimidine-7(8H)-one 16a

Compound 10f (150 mg, 0.55 mmol) was dissolved in dimethyl sulfoxide (3 mL), and N,N-diisopropylethylamine (212 mg, 1.64 mmol) and intermediate IN-5 (80 mg, 0.42 mmol) were added at room temperature ), heated to 80°C and reacted for 1 hour, TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 16a (140 mg, yield 75%) as a yellow solid.

The second step (R)-5-(4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-7-oxo-7, 8-Dihydropyrido[2,3-d]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate tert-butyl ester 16b

Compound 16a (140mg, 0.33mmol) and 1-tert-butoxycarbonyl-3,6-dihydro-2H-pyridine-5-boronic acid pinacol ester 6g (141mg, 0.46mmol) were dissolved in 1,4-dioxane In the ring (10 mL) and water (3 mL), Pd(dppf)Cl ₂ dichloromethane complex (35 mg, 0.043 mmol) and sodium carbonate (97 mg, 0.92 mmol) were added at room temperature, and the temperature was raised to 100 ° C under nitrogen protection and stirred After 1 hour, the reaction was completed by TLC. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 16b (200 mg, crude product) as a brown oil, which was directly used in the next step.

LC-MS: m/z=530.3[M+H] ⁺

The third step 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-carboxylate tert-butyl ester 16c

Compound 16b (200 mg, crude product) was dissolved in methanol (10 mL), palladium/carbon (50 mg, 10%) was added at room temperature, and the temperature was raised to 30° C. for 2 hours under a hydrogen atmosphere. TLC showed that the reaction of the starting materials was completed. The reaction solution was cooled to room temperature, filtered through a pad of celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain the title compound 16c (220 mg, crude product) as a yellow solid, which was directly used in the next step.

LC-MS: m/z=532.3[M+H] ⁺

The fourth step 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-(piperidin-3-yl) Pyrido[2,3-d]pyrimidin-7(8H)-one 16d

Compound 16c (220 mg, crude product) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for 2 hours. TLC showed that the reaction of the starting material was completed. The reaction solution was adjusted to basic by adding saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 16d (210 mg, crude product) as a yellow solid, which was directly used in the next step.

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

Compound 16d (210 mg, crude product) was dissolved in methanol (5 mL), aqueous formaldehyde solution (125 mg, 1.54 mmol, 37%) and palladium/carbon (70 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC showed the reaction of the starting materials. Finish. The reaction solution was filtered, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 16 (70 mg, four-step yield 48%) as a pale yellow solid.

LC-MS: m/z=446.3 [M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.28(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, 2H), 2.04-1.76(m, 3H), 1.66(d, J=6.8Hz, 3H), 1.64-1.56(m ,1H).(97.17%purity by HPLC)

Example 17

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

The first step 3-(2-ethoxy-2-oxyethyl) piperidine-1-carboxylate tert-butyl ester 17b

N-Boc-3-piperidineacetic acid 17a (5.0 g, 20.55 mmol) was dissolved in N,N-dimethylformamide (50 mL), potassium carbonate (8.5 g, 61.65 mmol) and ethyl iodide (4.8 g) were added , 30.82 mmol), and reacted at room temperature for 5 hours. The reaction solution was added with water, 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 obtain the title compound 17b (5.2 g, yield 93%) as a yellow oil.

The second step 3-(4-chloro-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylic acid tert-butyl Ester 17c

Compound 13a (1.13 g, 6.59 mmol) was dissolved in tetrahydrofuran (70 mL), compound 17b (2.32 g, 8.56 mmol) was added, cooled to -15°C, and lithium diisopropylamide (6.6 mL, 13.17 mmol, 2.0 mmol) was added dropwise. M), after dropping, reacted at -10°C for 5 hours, raised to room temperature and reacted overnight, TLC showed that the reaction of the raw materials was completed. The reaction solution was cooled to 0°C, quenched with saturated aqueous citric acid, extracted with ethyl acetate, 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 the crude product, which was slurried with petroleum ether. , filtered to give the title compound 17c (840 mg, 34% yield) as a yellow solid.

LCMS: m/z=323.1[M+H-56] ⁺

The third step (S)-3-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-7-oxo Substituted-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)piperidine-1-carboxylate 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] Pyrimidine-6-yl)piperidine-1-carboxylate tert-butyl ester 17d-2

Compound 17c (421 mg, 1.11 mmol) was dissolved in dimethyl sulfoxide (4 mL), and intermediate IN-5 (200 mg, 0.10 mmol) and N,N-diisopropylethylamine (430 mg, 3.33 mmol) were added at room temperature ), the temperature was raised to 100°C for 3 hours, and TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, extracted with ethyl acetate, the organic phase was washed with 1N hydrochloric acid, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 17d (310 mg, yield 52) as a yellow solid. %). Compound 17d was chiral resolved (Daicel AD-H, 30*250mm, 5μm, 30mL/min, IPA:Hexane=5:95) to obtain compound 17d-1 (peak No. 1, RT 7.95min) (100mg , yield 17%) and pale yellow solid compound 17d-2 (peak No. 2, RT 11.33 min) (114 mg, yield 20%). The configuration and properties of the compounds need to be further tested, and tentatively 17d-1 and 17d-2 are the above configurations.

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

The fourth step 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((S)-piperidine- 3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17e-1&4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl yl)amino)-8-methyl-6-((R)-piperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17e-2

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

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

The configuration and properties of the compound need to be further tested, and tentatively 17e-1 and 17e-2 are the above configurations.

The fifth step 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8-methyl-6-((S)-1-methyl ylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17-1&4-(((R)-1-(3-(difluoromethyl)-2-fluoro Phenyl)ethyl)amino)-8-methyl-6-((R)-1-methylpiperidin-3-yl)pyrido[2,3-d]pyrimidin-7(8H)-one 17 -2

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

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

The configuration and properties of the compound need to be further tested, and tentatively 17-1 and 17-2 are the above configurations.

17-1:

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

¹ H NMR (400MHz, CD ₃ OD) δ 8.29(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).

17-2:

LCMS: m/z=446.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.29(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).

Example 18

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

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

The compound of Example 6 was resolved by chirality (Daicel AD-H, 30*250mm, 5μm, 30mL/min, IPA:Hexane=5:95) to obtain solid title compounds 18-1 and 18-2. The configuration and properties of the compounds need to be further tested, and tentatively 18-1 and 18-2 are the above configurations.

18-1:

LC-MS: m/z=475.3 [M+H] ⁺

¹ H NMR(400MHz,Methanol-d ₄ )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).

18-2:

LC-MS: m/z=475.3 [M+H] ⁺

¹ H NMR(400MHz,Methanol-d ₄ )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=11.6 Hz, 2H), 1.94 (dd, J=19.9, 13.4Hz, 2H), 1.83 (t, J=12.8Hz, 1H), 1.60 (d, J=7.1Hz, 4H).

Example 19

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

The compound of Example 19 was synthesized with reference to the synthesis method of Example 20.

Example 20

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

The first step (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-carboxylate 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

Compound 17c (341 mg, 0.90 mmol) was dissolved in dimethyl sulfoxide (4 mL), and intermediate IN-8 (153 mg, 0.81 mmol) and N,N-diisopropylethylamine (348 mg, 2.70 mmol) were added at room temperature ), the temperature was raised to 100°C for 3 hours, and TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, extracted with ethyl acetate, the organic phase was washed with dilute hydrochloric acid (1N), washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 20a (198 mg, yield) as a yellow solid. rate 46%). Compound 20a was chiral resolved (Daicel AD-H, 30*250mm, 5μm, 30mL/min, IPA:Hexane=5:95) to obtain compound 20a-1 (peak No. 1, RT 8.75min) (85mg , yield 20%%) and pale yellow solid compound 20a-2 (peak No. 2, RT 8.53 min) (94 mg, yield 22%). The configuration and properties of the compounds need to be further tested, and tentatively 20a-1 and 20a-2 are the above configurations.

LCMS: m/z=532.2[M+H] ⁺

The second step 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

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

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

The configuration and properties of the compounds need to be further tested, and 20b-1 and 20b-2 are tentatively set as the above configurations.

The third step 8-methyl-6-((S)-1-methylpiperidin-3-yl)-4-(((R)-1-(3-(trifluoromethyl)phenyl)ethyl yl)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

Compound 20b-1 (100 mg, crude product) was dissolved in ethanol (5 mL), aqueous formaldehyde solution (100 mg, 1.23 mmol, 37%) and palladium/carbon (40 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC showed The reaction is complete. The reaction solution was filtered through a pad of celite, the filter cake was washed with methanol, and the filtrate was concentrated to obtain the title compound 20-1, 20-2 (55 mg, two-step yield 65%) as pale yellow solids.

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

The configuration and properties of the compound need to be further tested, and tentatively 20-1 and 20-2 are the above configurations.

20-1:

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

¹ H NMR (400MHz, CD ₃ OD) δ 8.32 (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).

20-2:

LCMS: m/z=446.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 8.32(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).

Example 21

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

Referring to Example 14, Example 21 was synthesized.

Example 22

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

The first step (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)thien-3-yl)benzyl)(methyl)carbamate 22a

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, and the temperature was raised to 100 ° C to react After 4 hours, TLC showed that the starting material had 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 obtain the title compound 22a (30 mg, yield 15%) as a white solid.

The second step 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

Compound 22a (30 mg, 0.047 mmol) was dissolved in methanol (5 mL), palladium/carbon (30 mg, 10%) was added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. TLC purification afforded the title compound 22 as a white solid (14 mg, 60% yield).

LC-MS: m/z=506.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 7.51 (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)

Example 23

2-Methyl-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 23

The first step 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)thiophen-3-yl)phenyl)carbamate 23a

Compound 2e (221 mg, 0.78 mmol) was dissolved in dimethyl sulfoxide (3 mL), and intermediate IN-2 (300 mg, 0.79 mmol) and N,N-diisopropylethylamine (356 mg, 2.75 mmol) were added at room temperature ), 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, 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 obtain the title compound 23a (140 mg, yield 29%) as a yellow solid. .

LC-MS: m/z=626.3[M+H] ⁺

The second step 2-methyl-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 23

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

LC-MS: m/z=492.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ 7.53(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,1H),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)

Example 24

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

The first step (2-((methylamino)methyl)phenyl)boronic acid 24a

o-formylboronic acid IN-2j (5.0g, 33.35mmol) was dissolved in methylamine ethanol solution (30mL, 7M), palladium carbon (200mg, 10%) was added, and the reaction was carried out at room temperature for 5 hours under a hydrogen atmosphere. TLC detected that the reaction of the raw materials was complete. . The reaction solution was filtered through a pad of celite, the filter cake was washed, and the filtrate was concentrated to obtain the title compound 24a (5.3 g, crude product) as a pale yellow foamy solid, which was directly used in the next step.

The second step (2-(((tert-butoxycarbonyl)(methyl)-14-azayl)methyl)phenyl)boronic acid 24b

Compound 24a (5.3 g, crude product) was dissolved in tetrahydrofuran (100 mL) and water (20 mL), sodium carbonate (10.3 g, 97.18 mmol) was added at room temperature, cooled to 0 °C, and di-tert-butyl dicarbonate (8.5 g) was added dropwise. , 38.95mmol), dripped, returned to room temperature and reacted overnight, and TLC detected that the reaction was complete. The reaction solution was added with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, slurried with crude petroleum ether and ethyl acetate (5/1), filtered, and the filter cake was washed and dried to obtain the title compound 24b (7.4 g, collected in two steps) as a white solid. rate 84%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.14 (s, 2H), 7.50 (dd, J=7.2, 0.8Hz, 1H), 7.38-7.30 (m, 1H), 7.21 (t, J=7.2 Hz, 1H), 7.08(d, J=7.6Hz, 1H), 4.56(s, 2H), 2.74(s, 3H), 1.40(s, 9H).

The third step (R)-(2-(5-(1-aminoethyl)thiophen-3-yl)benzyl)(methyl)carbamate tert-butyl ester 24c

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), and sodium carbonate (1.5 g, 14.15 mmol) and Pd(dppf)Cl ₂ dichloromethane complex (200 mg, 0.24 mmol), replaced with nitrogen for 3 times, heated to 100° C. for 4 hours, and the reaction was completed by TLC. The reaction solution was cooled to room temperature, water was added, 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 column chromatography to obtain the title compound 24c (800 mg, yield 48%) as a red oil. .

LC-MS: m/z=347.2[M+H] ⁺

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

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, cooled to -60 °C, and bistrimethylsilylamino was added dropwise Lithium (17.5mL, 0.175mol, 1.0M tetrahydrofuran solution) was dropped, and reacted at -10°C for 5 hours. TLC showed that the reaction of the intermediate was completed, and a small amount of raw materials were not fully reacted. The reaction was quenched by dropwise addition of saturated aqueous citric acid solution, adjusted to acidity, 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 obtain the title compound 24d as a brown-yellow solid (7.2 g, 36% yield).

LCMS: m/z=282.1[M+H] ⁺

The fifth step 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)thiophen-3-yl)benzyl)carbamate 24e

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, and the temperature was increased. The reaction was carried out at 90°C overnight, and TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, water was added, 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 obtain the title compound 24e (80 mg, yield 47%) as a yellow solid.

LCMS: m/z=592.3[M+H] ⁺

The sixth step 8-methyl-4-(((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-6- (((S)-Tetrahydrofuran-3yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 24

Compound 24e (80 mg, 0.14 mmol) was dissolved in hydrochloric acid methanol (3 mL, 4 N), and reacted at room temperature for 2 hours. TLC showed that the reaction was complete. The reaction was added dropwise with saturated aqueous sodium bicarbonate solution to adjust the pH to basic, 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 obtain the title compound 24 (45 mg) as a yellow solid. , the yield is 67%).

LCMS: m/z=492.2[M+H] ⁺

¹ H NMR (400MHz, CD ₃ OD) δ8.37(s, 1H), 7.53(s, 1H), 7.40-7.47(m, 1H), 7.39-7.26(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)

Inhibitory activity of the compound of test example 1 on SOS1

Experimental steps:

1. Compound treatment: prepare a compound with a final concentration of 400 times. For example, if the final concentration of the detection is 5uM, prepare it to a concentration of 400 times, that is, 2mM. The compound was serially diluted to the set number of concentration points using an automatic micropipette.

2. Transfer the compound to the 384-well reaction plate: Use the ultrasonic nanoliter liquid processing system to transfer 50nL of the above diluted compounds from the Echo 384-well plate to the 384-well reaction plate, and transfer 100% of the 50nL for both the negative and positive controls. DMSO.

3. Prepare and transfer 4x Tag1-SOS1 solution: Prepare 4x Tag1-SOS1 solution with Diluent provided in the kit (KRAS-G12C/SOS1 BINGDING ASSAY KIT (Cisbio, Cat. No. 63ADK000CB16PEG)), transfer 5ul to 384 In the well reaction plate, for the negative control wells, transfer 5ul of Diluent to replace the enzyme solution and centrifuge at 1000rpm for 1 minute.

4. Prepare 4x Tag2-KRAS G12C solution: Prepare 4x Tag2-KRAS G12C solution with Diluent provided in the kit, transfer 5ul to a 384-well reaction plate, and centrifuge at 1000rpm for 1 minute.

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

6. Reading: Read the data fluorescence signal value (Ex665/Em615) with a microplate reader Envision.

7. Inhibition rate calculation and IC50 fitting

Replicate the values from the plate reader, where the maximum value refers to the positive control reading and the minimum value refers to the negative control reading. Inhibition rate (%)=(maximum value-sample value)/(maximum value-minimum value)×100%.

Data were imported into MS Excel and _IC50 values were fitted with XLFit excel add-in version 5.4.0.8; results are shown in Table 1.

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

Inhibitory activity of the compounds provided in the examples of Table 1 on SOS1

化合物编号Compound number	IC ₅₀(nM) _IC50 (nM)	化合物编号Compound number	IC ₅₀(nM) _IC50 (nM)	化合物编号Compound number	IC ₅₀(nM) _IC50 (nM)
11	8.58.5	22	9.79.7	33	8.08.0
44	6.56.5	55	6.16.1	66	5.95.9
77	5.35.3	88	12.012.0	99	13.013.0
1010	5.05.0	1313	4.64.6	1414	5.75.7
17-117-1	5.15.1	17-217-2	9.39.3	18-118-1	4.94.9
18-218-2	1111	20-120-1	8.38.3	20-220-2	1515
22twenty two	8.08.0	23twenty three	5.05.0	24twenty four	5.85.8

IC ₅₀ Determination of the Compounds of Test Example 2 for Inhibition of K-562 Cell Proliferation

The human chronic myelogenous leukemia cell K-562 (CCL-243) used in the present invention was purchased from American Type Culture Collection (ATCC). Cells were grown in RPMI 1640 medium with 10% fetal bovine serum (FBS) and 1% dual antibodies at 37°C, 5% CO ₂ .

The inhibitory effect of compounds on the proliferation of K-562 cells cultured in vitro was determined by the following methods:

1) Cell seeding: Take K-562 cells with good logarithmic growth phase and seed them into a 96-well plate at 20,000 cells/well, 90 μL, and culture at 37° C. and 5% CO ₂ for 24 hours.

2) Dosing: Dilute the compound to be tested in a gradient in complete medium, and add 10 μL of the diluted compound to 90 μL of cells, so that the final concentration of the compound is 10000, 3000, 1000, 300, 100, 30, 10, 3 , 1nM, and set the corresponding solvent control at the same time. Place in a 37°C, 5% _CO2 cell incubator for 96 hours.

3) Detection: add 10 μL of 5 mg/mL MTT working solution (ABCONE, M9609) to each well, and after 4 hours at 37 °C, add triple solution (10% SDS, 0.5% isopropanol, 0.1 mol/L HCL) to the cells. The lysate was completely dissolved, and the OD570 and OD690 values were read using a TECAN SPARK microplate reader.

4) Calculation: Calculate the cell growth inhibition rate with the following formula:

Inhibition rate = (control well _{OD570nm-OD690nm} - administration well _{OD570nm-OD690nm} )/control well _{OD570nm-OD690nm} ×100%

_IC50 values were calculated from compound concentrations and corresponding inhibition ratios using Graphpad prism 5.0 software. The test results are shown in Table 2.

Table 2 IC ₅₀ (nM) of the compounds of the present invention for inhibition of proliferation of K-562 cells

化合物编号Compound number	K-562K-562	化合物编号Compound number	K-562K-562
BI-3406BI-3406	35.235.2	1313	99.999.9
11	461.7461.7	1515	17.017.0
22	246.9246.9	1616	38.338.3
33	118.3118.3	17-117-1	2828
44	99.999.9	17-217-2	148.8148.8
55	75.675.6	18-118-1	50.750.7
66	94.994.9	18-218-2	206.8206.8
88	288.8288.8	20-120-1	29.529.5
99	326.5326.5	20-220-2	198.3198.3
1010	34.034.0	22twenty two	333333
1111	37.137.1	23twenty three	274.3274.3

Conclusion: The compounds in the examples of the present invention can inhibit the proliferation of K-562 cells, and the activities of several compounds are comparable to BI-3402.

The effect of the compound of test example 3 on the phosphorylation level of KRAS downstream signaling molecule ERK1/2 in K-562 cells

The effect of the compounds of the present invention on the phosphorylation level of ERK1/2 in K-562 cells was determined by the following methods:

1) Cell seeding: K-562 cells with good logarithmic growth phase were seeded into a six-well plate at 1*10 ⁶ cells/well, and cultured overnight at 37° C. and 5% CO 2 .

2) Dosing: After the compound to be tested is serially diluted in complete medium, it is added to the cells so that the final concentration of the compound is 1000, 100, 10, and 1 nM. Place in a 37°C, 5% _CO2 cell incubator for 24 hours.

3) Protein sample preparation: collect the cell suspension, centrifuge at 500g for 5 minutes, discard the supernatant, wash 3 times with PBS, and use 1×SDS gel loading buffer (50mM Tris-HCl (pH 6.8), 100mM DTT, 2% SDS, 10% glycerol, 0.1% bromophenol blue) 100 μL of lysed cells. Cell lysates were denatured by heating at 100°C for 10 minutes.

4) Western blot: perform SDS-PAGE electrophoresis on the protein samples, after electrophoresis, transfer the protein to PVDF membrane with a wet transfer system, and place the PVDF membrane in blocking solution (5% nonfat milk powder diluted in TBS/T) to block at room temperature 1 hour, then I, II anti-reaction; after washing the membrane, use Immobilon Western HRP Substrate luminal reagent reagent to develop color, and take pictures with Western Blot imager (Tanon, 4600). The following is the antibody information used: p-ERK1/2 (CST:4370); ERK1/2 (CST:9102); β-tubulin (CST:2146).

The results of compound effects on ERK1/2 phosphorylation levels in K-562 cells are shown in Figure 1 .

Conclusion: Compound 5 in the examples of the present invention has a significant inhibitory effect on the phosphorylation of ERK1/2 in K-562 cells, and the inhibitory activity is concentration-dependent, and the activity is roughly equivalent to that of BI-3402.

Test Example 4 Compound Stability Experiment on Liver Microsomes in Mice and Humans

Experimental steps:

(1). Take out the liver microsomes (20 mg protein/mL) from the -80°C refrigerator, pre-incubate on a 37°C water bath thermostatic shaker for 3 min, and thaw for use.

(2). Prepare a mixed solution of the incubation system (without β-NADPH) according to the proportion of the above "Constitution of the experimental incubation system".

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

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

(5). Sample group: add 25 μL of β-NADPH solution (4 mM) to 75 μL of the reaction system described in (2), vortex for 30 s, mix well, and repeat the sample in a total reaction volume of 100 μL. Put it into a 37°C water bath constant temperature shaker for incubation, and start timing. The sampling time points are 0min, 5min, 15min, 30min, and 60min.

(6). Take out the sample tube at each time point, add 300 μL of cold stop agent (including internal standard), and stop the reaction.

(7). Vortex and centrifuge.

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

Data Analysis: Half-life (t _1/2 ) and clearance (CL) were calculated using the following first-order kinetic equations

C _t =C ₀ *e ^-kt

C _t =(1/2)*C ₀

t _1/2 =ln2/k=0.693/k

CL= _Vd *k

Vd=1/protein content in liver microsomes

CL _int(liver) = CL _int(mic) × liver weight to body weight ratio × liver microsomal protein concentration per gram of liver

The parameters in the formula are shown in Table 3:

Table 3 Common parameters of mouse, rat, and human liver and blood

The experimental results are shown in Table 4:

Table 4 Hepatic microsomal stability of compounds in different species

In the stability test of human liver microsomes, the stability of

compounds

2, 3, 4, and 13 was significantly better than that of BI-3406, and the stability of compound 5 was comparable to that of BI-3406; in the stability test of rat liver microsomes, The stability of

compounds

3, 5, 6, and 13 was significantly better than that of BI-3406, and the stability of compound 4 was comparable to that of BI-3406; in the stability test of mouse liver microsomes, compounds 2, 3, 4, 5, 6 ,13,18 The stability was significantly better than that of BI-3406. In conclusion, the stability of liver microsomes in different species of the compounds in the examples is significantly better than that of BI-3406.

CYP Enzyme Inhibition Study of the Compound of Test Example 5

Experimental steps:

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

(2) 100× compound: take the compound stock solution and dilute it with pure acetonitrile to 2000 μM working solution;

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

(4) use PB solution as solvent to prepare 4mM NADPH solution;

(5) Mix4in1 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 microsome (20mg/mL) solution, substrate working solution (Phe: 1A2 , 18 mM; Bup: 2B6, 12 mM; Amo: 2C8, 200 μM; Tes: 3A4, 4 mM), vortex and mix, and aliquot at 148 μL/tube.

(6) Mix3in1 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 microsome (20mg/mL) solution, substrate working solution (Dic: 2C9 , 1.6mM; DM: 2D6, 800 μM, Mid: 3A, 400 μM), vortex to mix, and aliquot at 148 μL/tube.

(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, and then add human liver microsome (20mg/mL) solution, the substrate works solution (Mep: 2C19, 4mM), mixed by vortex, and aliquoted at 148 μL/tube.

(8) Add 1 μL of analyte working solution/50% acetonitrile-water to each tube of the analyte, then add 1 μL of the analyte working solution/50% acetonitrile-water, and add 2 μL of the inhibitor working solution/50% acetonitrile-water to each tube of the inhibitor group. Pre-incubate in a 37°C water bath for 5 minutes, and at the same time pre-incubate NADPH in a 37°C water bath for 5 minutes;

(9) Add 50 μL/well of NADPH working solution, incubate for 30 min (S-mephenytoin), 10 min (other substrates);

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

(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 and mix, and inject by LC-MS/MS analyze.

data analysis

Use Excel to calculate the inhibition rate of the analyte and control on the metabolic rate of each specific probe substrate at different concentration levels, and then calculate the IC ₅₀ according to the following formula. The results are shown in Table 5:

* _IC50 =x*(100-enzyme inhibition % at x)/enzyme inhibition %, assuming Hillslope=1.

x is the inhibitor concentration

Table 5 Inhibitory activity of example compounds on CYP enzymes

Generally speaking, for each subtype of CYP450, IC ₅₀ >10μmol/L can be considered as weak inhibition of CYP; 3μmol/L<IC ₅₀ <10μmol/L can be considered as moderate inhibition of CYP; IC ₅₀ <3μmol/L can be considered as CYP Strong inhibition. It can be seen that BI-3406 exhibits moderate inhibitory activity in 2C9, 2C19, 2D6, 3A and other CYP450 subtypes. However, compounds 2, 3, 13, 18, 23, 24 showed weak inhibition of the 2C9 subtype; compounds 2, 3, 5, 6, 13, 18, 23, 24 showed weak inhibition of the 2C19 subtype;

compound

2 , 3, 4, 5, 6, 8, 13, 18 showed weak inhibition against 2D6 subtype; compounds 3, 6, 13, 18, 23, 24 showed weak inhibition against 3A subtype; tested compounds against 1A2, Both 2B6 and 2C8 showed weak inhibitory activity. In summary, it can be seen that the inhibition of CYP450 by multiple compounds is weaker than that of BI-3406, suggesting that it may have a lower risk of drug interactions and better drug safety.

In vivo antitumor activity of the compound of Test Example 6

Female BALB/c mice (6-8 weeks) were injected subcutaneously with MIAPaCa-2 or HPAF-II cells (1.0×10 ⁷ cells/mouse) on the right side. Mice were intragastrically administered with Compound 5 (50 mg/kg, bid), MRTX849 (5 mg/kg, qd), Trametinib (0.125 mg/kg, bid), and the combination of Compound 5 and Trametinib or MRTX849. Mice were monitored daily and caliper measurements were initiated when tumors became visible. The volume of the tumor was calculated by measuring two perpendicular diameters using the following formula: (L*W2)/2, where L and W refer to the length and width of the tumor diameter. When the mean tumor volume reached 100 ^mm3 , mice were grouped (D0, n=5/group) and administered compound. During the dosing period, tumor volume and mouse body weight were measured every 3 days. The results are shown in Figure 2-5.

Conclusion: The combination of compound 5 of the present invention and KRAS G12C inhibitor MRTX849 has a significant synergistic effect on MIAPaCa-2 (KRAS G12C) nude mice xenografts. kg, qd), the tumor inhibition rates of the single-drug group and the two-drug combination group were 71.2%, 60.8% and 98.1% (P<0.001). G12D) nude mice xenograft tumor has a significant synergy, the tumor inhibition rate of Example 5 (50mg/kg, bid) single-drug group, Trametinib (0.125mg/kg, bid) single-drug group and two-drug combination group was 61.3% , 76.2% and 105.3% (P<0.001). In conclusion, in the mouse model, the activity of the compound 5 of the example of the present invention alone is roughly equivalent to that of BI-3402, and the combination with MRTX849 or Trametinib has better antitumor activity against KRAS mutant tumors, which is significantly better than the respective single drug groups. .

The applicant declares that the present invention is to illustrate a pyrimidopyridone derivative of the present invention as an SOS1 inhibitor and its preparation method and application through the above-mentioned examples, but the present invention is not limited to the above-mentioned examples, that is, it does not mean that Therefore, the present invention can only be implemented by relying on the above-mentioned embodiments. Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

The preferred embodiments of the present invention are described in detail above, but the present invention is not limited to the specific details of the above-mentioned embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention. These simple modifications All belong to the protection scope of the present invention.

In addition, it should be noted that the specific technical features described in the above-mentioned specific embodiments can be combined in any suitable manner under the condition of no contradiction. In order to avoid unnecessary repetition, the present invention has The combination method will not be specified otherwise.

Claims

A pyrimidopyridone derivative, a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, wherein the structure of the pyrimidopyridone derivative is as shown in formula (I ) as shown:

wherein: R 1 is selected from hydrogen or C 1 -C 3 alkyl;

R 2 is selected from hydrogen or C 1 -C 3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein C 1 -C 3 alkyl, 3-7 membered cycloalkyl, 4-7 membered cycloalkyl A membered heterocyclyl group is optionally substituted with 1-3 R 21 ;

R 21 is selected from C 1 -C 3 alkyl, hydroxy, halogen, cyano, amino, C 1 -C 3 alkoxy or =O;

L may be absent or selected from O, NH or N-(C 1 -C 3 alkyl);

R 3 is selected from H, C 1 -C 3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein C 1 -C 3 alkyl, 3-7 membered cycloalkyl, 4-7 membered cycloalkyl A membered heterocyclyl group is optionally substituted with 1-3 R 31 ;

R 31 is selected from C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, hydroxy, halogen, cyano, -NR a R b , C 1 -C 3 alkoxy, =O, -NHCOR 32 or - COR 32 ;

R a is selected from H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl or 3-6 membered cycloalkyl;

R b is selected from H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl or 3-6 membered cycloalkyl;

R 32 is selected from C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclyl;

AR is selected from 6-10-membered aryl or 5-10-membered heteroaryl, wherein the aryl or heteroaryl is optionally substituted by 1-4 R 4 ;

R 4 is selected from H, halogen, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, hydroxy-C 1 -C 3 alkyl, hydroxy-C 1 -C 3 haloalkyl, 3-6 membered cycloalkane base, 4-7 membered heterocyclic group, -OR a , -NR a R b , 6-10 membered aryl group or 5-10 membered heteroaryl group, of which 6-10 membered aryl group or 5-10 membered aryl group The heteroaryl group is optionally substituted by 1-4 R c ;

R c is selected from H, halogen, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, hydroxy-C 1 -C 3 alkyl, hydroxy-C 1 -C 3 haloalkyl, 3-6 membered cycloalkane base, 4-7 membered heterocyclic group, -OR a , -NR a R b , NR a R b -C 1 -C 4 alkyl, NR a R b -C 1 -C 4 haloalkyl; the formula ( The heteroatoms in the heterocyclic group or heteroaryl group in I) are 1-3 and are selected from one or more of oxygen, nitrogen and sulfur.
The pyrimidopyridone derivatives, pharmaceutically acceptable salts thereof, tautomers or stereoisomers thereof according to claim 1, wherein the pyrimidopyridone derivatives have The structure is shown in formula (II):

Wherein, R 1 , R 2 , R 3 and R 4 have the same limited range as claim 1; n=1-4;

Preferably, in the compound of formula (II), the phenyl group is optionally substituted by 1-4 R 4 , when the number of the R 4 is 2-4, the R 4 can be the same or different;

And/or, when the R 4 is a C 1 -C 3 haloalkyl group, the R 4 is 1-3, and when there are two or more R 4 , the R 4 can be the same or different;

And/or, when the R 4 is a C 1 -C 3 haloalkyl group, the halogen atom therein is fluorine;

And/or, when the R 4 is halogen, the R 4 is 1-3, and when there are two or more R 4 , the R 4 can be the same or different;

And/or, when the R 4 is halogen, the halogen atom is fluorine;

And/or, when the R 4 is -NR a R b , the R 4 is 1-3, and when there are two or more R 4 , the R 4 can be the same or different;

And/or, when the R 4 is -NR a R b , the R a and R b can be the same or different;

and/or R 3 is H, C 1 -C 3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein the 4-7 membered heterocyclyl is optionally surrounded by 1-3 R 31 replaced;

And/or, when R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , and when said R 31 is 2-3, R 31 is the same or different;

And/or, when the R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , the heterocyclic group contains 1-2 heteroatoms;

And/or, when the R 3 is a 4-7 membered heterocyclic group optionally substituted by 1-3 R 31 , the heteroatom of the heterocyclic group is nitrogen and/or oxygen;

And/or, when the R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , when the heteroatoms of the heterocyclic group are two, the two hetero atoms are the same or different;

And/or, when the R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , the R 31 is selected from C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, hydroxy, halogen, cyano, -NR a R b , C 1 -C 3 alkoxy, =O, -NHCOR 32 or -COR 32 ;

R a is selected from H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl or 3-6 membered cycloalkyl;

R b is selected from H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl or 3-6 membered cycloalkyl;

R 32 is selected from C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclyl.
The pyrimidopyridone derivatives, pharmaceutically acceptable salts thereof, tautomers or stereoisomers thereof according to claim 1, wherein the pyrimidopyridone derivatives have The structure is shown in formula (III):

Wherein, R 1 , R 2 , R 3 and R 4 have the same limited range as claim 1; n=1-4;

Preferably, for the compound of formula (III), the phenyl group is optionally substituted by 1-4 R 4 , when the number of the R 4 is 2-4, the R 4 can be the same or different;

And/or, when the R 4 is a C 1 -C 3 haloalkyl group, the R 4 is 1-3, and when there are two or more R 4 , the R 4 can be the same or different;

And/or, when the R 4 is a C 1 -C 3 haloalkyl group, the halogen atom therein is fluorine;

And/or, when the R 4 is halogen, the R 4 is 1-3, and when there are more than two R 4 , the R 4 can be the same or different;

And/or, when the R 4 is halogen, the halogen atom is fluorine;

And/or, when the R 4 is -NR a R b , the R 4 is 1-3, and when there are more than two R 4 , the R 4 can be the same or different;

And/or, when the R 4 is -NR a R b , the R a and R b can be the same or different;

and/or R 3 is H, C 1 -C 3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclyl, wherein the 4-7 membered heterocyclyl is optionally surrounded by 1-3 R 31 replaced;

And/or, when R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , and when said R 31 is 2-3, R 31 is the same or different;

And/or, when the R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , the heterocyclic group contains 1-2 heteroatoms;

And/or, when the R 3 is a 4-7 membered heterocyclic group optionally substituted by 1-3 R 31 , the heteroatom of the heterocyclic group is nitrogen and/or oxygen;

And/or, when the R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , when the heteroatoms of the heterocyclic group are two, the two hetero atoms are the same or different;

And/or, when the R 3 is a 4-7-membered heterocyclic group optionally substituted by 1-3 R 31 , the R 31 is selected from C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, hydroxy, halogen, cyano, -NR a R b , C 1 -C 3 alkoxy, =O, -NHCOR 32 or -COR 32 ;

R a is selected from H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl or 3-6 membered cycloalkyl;

R b is selected from H, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl or 3-6 membered cycloalkyl;

R 32 is selected from C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclyl.
The pyrimidopyridone derivatives, pharmaceutically acceptable salts thereof, tautomers or stereoisomers thereof according to claim 1, wherein the pyrimidopyridone derivatives have The structure is shown in formula (IV):

wherein, R 1 , R 2 , R 3 , R c and L have the same limited range as claim 1; m=1-4;

Preferably, in the compound of formula (IV), L is selected from O, NH or NCH 3 .
The pyrimidopyridone derivatives, pharmaceutically acceptable salts thereof, tautomers or stereoisomers thereof according to claim 1, wherein the pyrimidopyridone derivatives are selected from from any of the following structures:
A preparation method of a pyrimidopyridone derivative as claimed in any one of claims 1-5, its stereoisomer, its tautomer or its pharmaceutically acceptable salt, which is selected from the following two One of the methods:

Method 1. The compound of general formula (I-B) and the compound of general formula (I-A) are subjected to substitution reaction to obtain the compound of general formula (I):

Wherein, X 1 is halogen, preferably chlorine; R 1 , R 2 , R 3 , AR and L have the same limited scope as in claim 1 .

Method Two,

by compound
and compound R 3 W under basic conditions, in the presence of metal catalyst and ligand, through Suzuki reaction to obtain the compound of general formula (I);

Wherein, X 4 is halogen, preferably bromine; W is
R 1 , R 2 , R 3 , AR and L have the same limited scope as claim 1 .
A pharmaceutical composition, characterized in that the pharmaceutical composition comprises the pyrimidopyridone derivatives according to any one of claims 1-5, their stereoisomers, their tautomers or pharmaceutically acceptable salts thereof;

Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and/or excipient.
The pyrimidopyridone derivatives according to any one of claims 1 to 5, their stereoisomers, their tautomers, their pharmaceutically acceptable salts, or the pharmaceutical composition according to claim 7 Use alone or in combination with a KRAS inhibitor or MEK signaling pathway inhibitor in the preparation of a medicament for the treatment of cancer or in the preparation of an SOS1 inhibitor; preferably, the cancer includes but is not limited to astrocytic carcinoma, breast cancer, cervical cancer cancer, colorectal cancer, endometrial cancer, esophagus, stomach, head and neck, hepatocellular, laryngeal, pancreatic, lung, oral, ovarian, prostate, thyroid, sarcoma, kidney and Cholangiocarcinoma; further preferably, the cancer includes but is not limited to: pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, kidney cancer, gastric cancer and cholangiocarcinoma; and/or the KRAS inhibitor is KRAS G12C, KRAS G12V , KRAS G12S or KRAS G12D inhibitor; and/or said MEK signaling pathway inhibitor such as RAF, MEK or ERK1/2 inhibitor.
The use according to claim 8, the dosage range of the pyrimidopyridone derivatives, their stereoisomers, their tautomers, their pharmaceutically acceptable salts or pharmaceutical compositions is 10-100 mg/kg and/or the dose range of KRAS G12C inhibitor is selected from 3-100mg/kg; and/or the dose range of MEK inhibitor is selected from 0.1-0.2mg/kg; preferably, the pyrimidopyridone derivatives, its stereoisomer, its tautomer, its pharmaceutically acceptable salt or pharmaceutical composition in a dosage range of 25-50 mg/kg; and/or a KRAS G12C inhibitor in a dosage range of 5-30 mg/kg; and and/or the KRAS G12C inhibitor is MRTX849, and/or the MEK inhibitor is Trametinib.
Use according to claim 8, wherein the cancer is a RAS family related cancer; preferably a KRAS, HRAS or NRAS G12C related cancer, KRAS, HRAS or NRAS G12D related cancer, KRAS, HRAS or NRAS G12V related cancer, KRAS, HRAS or NRAS G12S-related cancer, KRAS, HRAS or NRAS G12A-related cancer, KRAS, HRAS or NRAS G13D-related cancer, KRAS, HRAS or NRAS G13C-related cancer, KRAS, HRAS or NRAS Q61L-related cancer, KRAS, HRAS or NRAS A146T-related cancer, KRAS, HRAS or NRAS A146V-associated cancer or KRAS, HRAS or NRAS A146P-associated cancer.
A method for the prevention and/or treatment of cancer, comprising administering to a human a therapeutically effective amount of a compound according to any one of claims 1-5, or a pharmaceutically acceptable compound thereof, a stereoisomer thereof, or a compound thereof. A variant, a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 7.