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

Abstract

The present invention belongs to the technical field of polycyclic pyrimidine derivatives, and particularly relates to a polycyclic pyrimidine derivative as an SOS1 inhibitor, a preparation method thereof, and uses thereof. The structure of the polycyclic pyrimidine derivative is shown in formula (I). The present invention provides a novel polycyclic pyrimidine derivative, which has a significant effect of inhibiting the RAS signaling pathway and treating cancers, and the cancers include pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer, cholangiocarcinoma, etc.

Description

Technical Field

[0001] The present invention belongs to the technical field of polycyclic pyrimidine derivatives, and particularly relates to a polycyclic pyrimidine derivative as an SOS1 inhibitor, a preparation method thereof, and uses thereof. Background Art

[0002] RAS family proteins belong to a type of small GTPase and include three subfamilies: KRAS, NRAS, and HRAS. The mutated RAS gene is an important oncogene, and the discovery of RAS gene mutations exists in 20 - 30% of human tumors, especially pancreatic cancer, colorectal cancer, and lung cancer. There is a balance between the GTP-bound active state and the GDP-bound inactive state for various subtypes of RAS proteins. GTPase-activating proteins (GAPs) can promote the conversion of GTP to GDP, thus causing the RAS protein to shift to the inactive state, while guanine nucleotide exchange factors (GEFs) can promote the release of GDP and the binding of GTP, thus causing the RAS protein to shift to the active state. The activation of the RAS protein promotes cell proliferation, apoptosis evasion, and metabolic reorganization, etc. through the RAS-RAF-MEK-ERK and RAS-PI3K-PDK1-AKT signaling pathways, thereby promoting the occurrence and development of tumors.

[0003] SOS1 (son of sevenless 1) is a key guanine nucleotide exchange factor (GEF) that can bind to the RAS protein, promote the binding of the RAS protein to GTP, and cause the RAS protein to shift to the active state. Recent studies have found that inhibitors of SOS1 can not only inhibit the growth of RAS mutant cells but also produce a synergistic effect with MEK inhibitors, showing a significant inhibitory effect on KRAS-driven tumors. 1-2 The development of SOS1 inhibitors has become a research hotspot. Different structural types of SOS1 inhibitors have been reported in many patents, such as WO2018172250, WO2019201848, WO2018115380, WO2019122129, WO2020173935, WO2020180768, and WO2020180770, etc.

[0004] However, the compounds and experimental drugs disclosed in these prior arts still have uncertainties in terms of effectiveness, safety, or selectivity, etc. Therefore, it is necessary to research and develop new selective SOS1 inhibitors.

[0005] References:

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

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

[0008] In order to solve the above problems of the prior art, the object of the present invention is to provide a polycyclic pyrimidine derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, so as to screen out a compound used as an SOS1 inhibitor that has excellent properties in terms of effectiveness, safety, selectivity and other performances.

[0009] Another object of the present invention is to provide a preparation method of the derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof.

[0010] To achieve the object of the present invention, the following technical solutions are adopted:

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

[0012]

[0013] Wherein: R 1 is selected from hydrogen or C1 - C3 alkyl; preferably hydrogen or methyl;

[0014] A 1 is selected from N or C - R 11 ;

[0015] R 11 is selected from H, C1 - C3 alkyl or C1 - C3 haloalkyl;

[0016] A 2 is selected from N or C - R 2 :

[0017] R 2 Selected from -OR 21 or cyano;

[0018] R 21 Selected from H, C1-C3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclic group, wherein the C1-C3 alkyl, 3-7 membered cycloalkyl, 4-7 membered heterocyclic group is optionally substituted by 1-3 R 22 substituted;

[0019] R 22 Selected from halogen, C1-C4 alkyl, cyano, hydroxy;

[0020] L may be absent or selected from O, NH or NCH3;

[0021] R 3 Selected from 3-7 membered cycloalkyl, 4-7 membered heterocyclic group, 6-10 membered fused heterocyclic group, 6-10 membered bridged heterocyclic group, 6-10 membered spiro heterocyclic group, wherein the 3-7 membered cycloalkyl, 4-7 membered heterocyclic group, 6-10 membered fused heterocyclic group, 6-10 membered bridged heterocyclic group, 6-10 membered spiro heterocyclic group is optionally substituted by 1-3 R 31 substituted;

[0022] R 31 Selected from C1-C3 alkyl, C1-C3 haloalkyl, hydroxy, halogen, cyano, -NR a R b , C1-C3 alkoxy, =O, -NHCOR 32 or -COR 32 :

[0023] R 32 Selected from C1-C3 alkyl, C1-C3 haloalkyl, 3-7 membered cycloalkyl or 4-7 membered heterocyclic group;

[0024] R 4 is -CH3;

[0025] 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 5 substituted;

[0026] R 5 Selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl, hydroxy-C1-C4 alkyl, hydroxy-C1-C4 haloalkyl, 3-6 membered cycloalkyl, 4-7 membered heterocyclic group, -OR a , -NR a R b ;

[0027] R aSelected from H, C1-C4 alkyl, C1-C4 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclic group;

[0028] R b Selected from H, C1-C4 alkyl, C1-C4 haloalkyl, 3-6 membered cycloalkyl or 4-7 membered heterocyclic group;

[0029] In the above definitions, the following conditions cannot occur simultaneously:

[0030] 1) A 1 is C-R 11 ;

[0031] 2) A 2 is C-OR 21 ;

[0032] 3) L is O or absent;

[0033] The heteroatoms in the heterocyclic group and heteroaryl in formula (I) are 1-3 and are selected from one or more of oxygen, nitrogen or sulfur.

[0034] Preferably, the structure of the polycyclic pyrimidine derivative is as shown in formula (II):

[0035]

[0036] wherein, R 1 , L, R 3 and R 5 have the same defined ranges as above; n = 1-4 (for example, n = 1, n = 2, n = 3, n = 4);

[0037] Preferably, for the compound of formula (II), the phenyl group is optionally substituted by 1-4 R 5 . When the number of R 5 is 2-4, the R 5 can be the same or different;

[0038] and / or, when the R 5 is C1-C4 haloalkyl, the R 5 is 1-3. When there are 2-3 R 5 , the R 5 can be the same or different;

[0039] and / or, when the R 5 is C1-C4 haloalkyl, the halogen atom therein is fluorine or chlorine;

[0040] and / or, when the R 5 is halogen, the R 5 is 1-3. When there are 2-3 R5 When, the R 5 may be the same or different;

[0041] And / or, when the R 5 is halogen, the halogen atom is fluorine or chlorine;

[0042] And / or, when the R 5 is -NR a R b When, the R 5 is 1 - 3. When there are 2 - 3 R 5 When, the R 5 may be the same or different;

[0043] And / or, when the R 5 is -NR a R b When, the R a and R b may be the same or different.

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

[0045]

[0046] Wherein, R 1 , R 21 , R 3 and R 5 have the same defined range as above; n = 1 - 4 (for example, n = 1, n = 2, n = 3, n = 4);

[0047] Preferably, for the compound of formula (III), when R 3 is a 4 - 7 - membered heterocyclic group optionally substituted by 1 - 3 R 31 , the 4 - 7 - membered heterocyclic group is a 5 - 6 - membered heterocyclic group;

[0048] And / or, when R 3 is a 4 - 7 - membered heterocyclic group optionally substituted by 1 - 3 R 31 , the heterocyclic group contains 1 - 2 heteroatoms;

[0049] 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;

[0050] And / or, when the R 3 is a 4 - 7 - membered heterocyclic group optionally substituted by 1 - 3 R 31 , when there are two heteroatoms in the heterocyclic group, the two heteroatoms are the same or different;

[0051] and / or, said R 21 is selected from H or optionally C1-C3 alkyl substituted by 1-3 R 22 , said R 22 is selected from halogen, C1-C4 alkyl, cyano, hydroxy;

[0052] wherein, R 31 and R 22 have the same defined scope as above.

[0053] Preferably, the structure of the polycyclic pyrimidine derivative is shown in formula (IV):

[0054]

[0055] wherein, R 1 , L, R 3 and R 5 have the same defined scope as above; n = 1-4 (for example, n = 1, n = 2, n = 3, n = 4);

[0056] Preferably, for the compound of formula (IV), when L is absent or is O, R 3 is 3-7 membered cycloalkyl or 4-7 membered heterocyclic group optionally substituted by 1-3 R 31 ;

[0057] and / or, when said R 3 is 4-7 membered heterocyclic group optionally substituted by 1-3 R 31 , the heteroatoms in the heterocyclic group are nitrogen and / or oxygen;

[0058] and / or, when said R 3 is 4-7 membered heterocyclic group optionally substituted by 1-3 R 31 , the number of heteroatoms is 1-2;

[0059] wherein, R 31 has the same defined scope as above.

[0060] Preferably, the structure of the polycyclic pyrimidine derivative is shown in formula (V):

[0061]

[0062] wherein, R 1 , A 2 , L, R 3 and R 5 have the same defined scope as above; n = 1-4 (for example, n = 1, n = 2, n = 3, n = 4).

[0063] Further preferably, the polycyclic pyrimidine derivatives are selected from any one of the following structures:

[0064]

[0065]

[0066]

[0067] Typical compounds of the present invention include, but are not limited to, the compounds in the following table:

[0068] In a second aspect, the present invention provides a method for preparing a polycyclic pyrimidine derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof as described in the first aspect, which is selected from one of the following four schemes:

[0069] Scheme 1

[0070] A method for preparing the compound of general formula (I) described in the present invention or its stereoisomer, tautomer or pharmaceutically acceptable salt thereof, includes the following steps:

[0071] Preparation of the key intermediate (I-A), Method 1:

[0072]

[0073] In the first step, the compound of general formula (I-1) undergoes metal-catalyzed cross-coupling to obtain the compound of general formula (I-2);

[0074] In the second step, the compound of general formula (I-2) reacts under catalyst conditions to obtain the chiral sulfonimide compound of general formula (I-3);

[0075] In the third step, the compound of general formula (I-3) is reduced by a metal reducing agent to obtain the chiral compound of general formula (I-4);

[0076] In the fourth step, the sulfonamide of the compound of general formula (I-4) is cleaved under acidic conditions to obtain the chiral benzylamine compound of general formula (I-A).

[0077] Preparation of the key intermediate (I-A), Method 2:

[0078]

[0079] In the first step, the alcoholic hydroxyl group of the compound of general formula (I-5) is oxidized to obtain the aldehyde compound of general formula (I-6);

[0080] In the second step, the compound of general formula (I-6) is subjected to a Grignard reaction to obtain the compound of general formula (I-7).

[0081] In the third step, the alcohol hydroxyl group of the compound of general formula (I-7) is oxidized to obtain the chiral compound of general formula (I-2).

[0082] The fourth to sixth steps are the same as above.

[0083] Among them, AR, R 4 has the same defined range as above.

[0084] Preparation of the key intermediate (I-B), Method 1:

[0085]

[0086] In the first step, the methyl group of the compound of general formula (I-8) is oxidized to obtain the acid compound of general formula (I-9).

[0087] In the second step, the compound of general formula (I-9) is subjected to an esterification reaction to obtain the compound of general formula (I-10).

[0088] In the third step, the compound of general formula (I-10) and the compound of general formula (I-11) react under basic conditions to obtain the compound of general formula (I-12).

[0089] In the fourth step, the nitro group of the compound of general formula (I-12) is reduced to obtain the compound of general formula (I-13).

[0090] In the fifth step, the compound of general formula (I-13) reacts with nitrile to form an amidine under acidic conditions and then cyclizes to obtain the compound of general formula (I-B).

[0091] Among them, A 1 、A 2 、R 1 、R 3 and L have the same defined range as above; A 1 is preferably C; A 2 is preferably N; L is preferably absent; R 1 is preferably methyl.

[0092] Preparation of the key intermediate (I-B), Method 2:

[0093]

[0094] In the first step, the compound of general formula (I-9) is subjected to acyl chlorination and reacts with ammonia water to obtain the amide compound of general formula (I-14).

[0095] In the second step, the compound of general formula (I-14) and the compound of general formula (I-11) react under basic conditions to obtain the compound of general formula (I-15).

[0096] In the third step, the nitro group of the compound of general formula (I-15) is reduced to obtain the compound of general formula (I-16).

[0097] In the fourth step, the compound of general formula (I-16) reacts with triethyl orthoacetate to obtain the compound of general formula (I-B) (R 1 = methyl); the compound of general formula (I-16) reacts with triethyl orthoformate to obtain the compound of general formula (I-B) (R 1 = H).

[0098] Wherein, A 1 、A 2 、R 1 、R 3 and L have the same defined ranges as above; A 1 is preferably C; A 2 is preferably N; L is preferably absent or O; R 1 is preferably methyl.

[0099] Preparation of the key intermediate (I-C), Method 1:

[0100]

[0101] In the first step, the compound of general formula (I-B) undergoes a halogenation reaction to obtain the compound of general formula (I-C).

[0102] Wherein, A 1 is preferably C, A 2 is preferably N, L is preferably absent or O, R 1 is preferably methyl.

[0103] Preparation of the key intermediate (I-C), Method 2:

[0104]

[0105] In the first step, the compound of general formula (I-11) and tert-butyl bromoacetate undergo a substitution reaction under basic conditions to obtain the compound of general formula (I-17);

[0106] In the second step, the compound of general formula (I-17) undergoes de-tert-butylation under acidic conditions to obtain the compound of general formula (I-18);

[0107] In the third step, the compound of general formula (I-18) undergoes an esterification reaction with an alcohol under acidic conditions to obtain the compound of general formula (I-19);

[0108] In the fourth step, the compound of general formula (I-20) reacts with ammonia water to obtain the compound of general formula (I-21);

[0109] In the fifth step, the compound of general formula (I-21) is subjected to a halogenation reaction to obtain the compound of general formula (I-22);

[0110] In the sixth step, the compound of general formula (I-22) and potassium vinyltrifluoroborate react under basic conditions in the presence of a metal catalyst and a ligand to obtain the compound of general formula (I-23) through a Suzuki reaction;

[0111] In the seventh step, the double bond in the compound of general formula (I-23) is oxidized to obtain the compound of general formula (I-24);

[0112] In the eighth step, the compound of general formula (I-24) and the compound of general formula (I-19) react under basic conditions to obtain the compound of general formula (I-25);

[0113] In the ninth step, the hydroxyl group of the compound of general formula (I-25) is substituted to obtain the compound of general formula (I-C).

[0114] Among them, A 1 , A 2 , R 1 , R 3 , and L have the same defined ranges as above; A 1 is preferably N; A 2 is preferably CR 2 (R 2 selected from -OR 21 ); L is preferably NH, and R 1 is preferably methyl.

[0115] Preparation of general formula (I):

[0116] The preparation method of the compound of general formula (I) described in the present invention or its stereoisomer, tautomer or its medicinal salt includes the following steps:

[0117]

[0118] In the first step, the compound of general formula (I-B) and the compound of general formula (I-A) react under condensation conditions to obtain the compound of general formula (I).

[0119] Among them, A 1 , A 2 , AR, R 1 , R 3 , R 4 and L have the same defined ranges as above.

[0120] Scheme 2

[0121] The preparation method of the compound of general formula (I) described in the present invention or its stereoisomer, tautomer or its medicinal salt includes the following steps:

[0122]

[0123] In the first step, a compound of formula (I-C) and a compound of formula (I-A) react under basic conditions to obtain a compound of formula (I).

[0124] wherein A 1 、A 2 、AR, R 1 、R 3 、R 4 and L are as defined in formula (I).

[0125] Scheme III

[0126] A method for preparing a compound of formula (I) described in the present invention, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, comprises the following steps:

[0127]

[0128] In the first step, a compound of formula (I-26) is demethylated under basic conditions to obtain a compound of formula (I-27);

[0129] In the second step, a compound of formula (I-27) is subjected to an esterification reaction to obtain a compound of formula (I-28);

[0130] In the third step, a protecting group is introduced onto the hydroxyl group of a compound of formula (I-28) to obtain a compound of formula (I-29);

[0131] In the fourth step, the nitro group of a compound of formula (I-29) is reduced to obtain a compound of formula (I-30);

[0132] In the fifth step, a compound of formula (I-30) reacts with a nitrile under acidic conditions to form an amidine and then cyclizes to obtain a compound of formula (I-31);

[0133] In the sixth step, the hydroxyl group of a compound of formula (I-31) is halogenated to obtain a compound of formula (I-32);

[0134] In the seventh step, a compound of formula (I-32) and a compound of formula (I-A) react under basic conditions to obtain a compound of formula (I-33);

[0135] In the eighth step, the protecting group of a compound of formula (I-33) is removed under acidic conditions to obtain a compound of formula (I-34);

[0136] In the ninth step, a compound of formula (I-34) reacts with trifluoromethanesulfonic anhydride under basic conditions to obtain a compound of formula (I-35);

[0137] In the tenth step, the compound of general formula (I-35) and the compound of general formula (I-11) are reacted in the presence of a metal catalyst and a ligand under basic conditions to obtain the compound of general formula (I).

[0138] Wherein, PG is a protecting group, preferably benzyl; A 1 、A 2 、AR、R 1 、R 3 、R 4 and L have the same defined ranges as above; L is preferably NH; A 1 is preferably C; A 2 is preferably CR 2 (R 2 is preferably -OR 21 ).

[0139] Scheme Four

[0140] The preparation method of the compound of general formula (I) described in the present invention, or its stereoisomer, tautomer or its medicinal salt, comprises the following steps:

[0141]

[0142] In the first step, the methyl group of the compound of general formula (I-36) is oxidized to obtain the acid compound of general formula (I-37);

[0143] In the second step, the compound of general formula (I-37) is subjected to a nitration reaction to obtain the compound of general formula (I-38);

[0144] In the third step, the compound of general formula (I-38) and the compound of general formula (I-11) are reacted under basic conditions to obtain the compound of general formula (I-39);

[0145] In the fourth step, the compound of general formula (I-39) is subjected to acyl chlorination and reaction with ammonia water to obtain the amide compound of general formula (I-40);

[0146] In the fifth step, the nitro group of the compound of general formula (I-40) is reduced to obtain the compound of general formula (I-41);

[0147] In the sixth step, the compound of general formula (I-41) is reacted with triethyl formate to obtain the compound of general formula (I-42) (R 1 =H);

[0148] In the seventh step, the compound of general formula (I-42) is subjected to a halogenation reaction to obtain the compound of general formula (I-43);

[0149] In the eighth step, the compound of general formula (I-43) and the compound of general formula (I-A) are reacted under basic conditions to obtain the compound of general formula (I-44);

[0150] In the ninth step, the compound of general formula (I-44) reacts with zinc cyanide in the presence of a metal catalyst to obtain the compound of general formula (I).

[0151] Among them, X 1 、X 2 and X 3 are halogens; X 1 is preferably fluorine; X 2 is preferably bromine; X 3 is preferably chlorine; A 1 、A 2 、AR、R 1 、R 3 、R 4 and L have the same defined ranges as above; A 1 is preferably C; A 2 is CR 2 (R 2 is preferably cyano); L is preferably O; R 1 is preferably methyl.

[0152] For the above preparation method,

[0153] The reagent providing the basic condition is selected from organic bases or inorganic bases. The organic bases are one or more of triethylamine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis(trimethylsilyl)amide, sodium tert-butoxide, sodium methoxide and potassium tert-butoxide. The inorganic bases are one or more of sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium bicarbonate and lithium hydroxide;

[0154] The reagent providing the acidic condition is one or more of hydrogen chloride, 1,4-dioxane solution of hydrogen chloride, methanol solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid and phosphoric acid;

[0155] The metal catalyst is one or more of palladium / carbon, Raney nickel, tetrakis(triphenylphosphine)palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (Pd(dppf)Cl2), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex, bis(triphenylphosphine)palladium dichloride (Pd(PPh3)Cl2) and tris(dibenzylideneacetone)dipalladium (Pd2(dba)3);

[0156] The ligand is one or more of 2-dicyclohexylphosphino-2,6'-dimethoxybiphenyl (SPhos), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (XantPhos), 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (XPhos), 2-dicyclohexylphosphino-2'-(N,N-dimethylamino)-biphenyl (DavePhos), 1,1'-bis(diphenylphosphino)ferrocene (Dppf), and 1,1'-binaphthalene-2,2'-bis(diphenylphosphine) (BINAP), preferably 1,1'-binaphthalene-2,2'-bis(diphenylphosphine) (BINAP);

[0157] The reducing agent is one or more of sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, and lithium aluminum hydride;

[0158] The oxidizing agent is one or more of potassium permanganate, manganese dioxide, potassium dichromate, sodium dichromate, and potassium osmate;

[0159] The above reaction is preferably carried out in a solvent, and the solvents used are one or more of N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, 1,4-dioxane, water, tetrahydrofuran, dichloromethane, 1,2-dichloroethane, methanol, ethanol, toluene, petroleum ether, ethyl acetate, n-hexane, and acetone.

[0160] In a third aspect, the present invention provides a pharmaceutical composition, which comprises the polycyclic pyrimidine derivatives, pharmaceutically acceptable salts thereof, tautomers thereof, and stereoisomers thereof as described in the first aspect; preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0161] In the present invention, the term "comprising" means that various components can be applied together in the mixtures or compositions of the present invention. Therefore, the terms "consisting essentially of..." and "consisting of..." are included in the term "comprising".

[0162] In the present invention, a "pharmaceutically acceptable" component is a substance that is suitable for humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, a substance with a reasonable benefit / risk ratio.

[0163] In the present invention, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent, or excipient used to deliver the active substance or its physiologically acceptable salt of the present invention to animals or humans. The carrier can be liquid or solid.

[0164] 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, even more preferably 0.1 - 90 parts by weight) of the compound of formula (I) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 parts by weight.

[0165] Alternatively, the pharmaceutical composition of the present invention contains 0.001 - 99.9 wt%, more preferably 0.01 - 99 wt%, even more preferably 0.1 - 90 wt% of the total weight of the compound of formula (I) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 wt%.

[0166] In another preferred embodiment, the preferred ratio of the compound of formula (I) to a pharmaceutically acceptable carrier, excipient or sustained release agent is such that the compound of formula (I) as the active ingredient accounts for more than 65% of the total weight, and the remaining part accounts for 0.5 - 40% of the total weight, or more preferably 1 - 20%, or most preferably 1 - 10%.

[0167] For various dosage forms of the pharmaceutical composition of the present invention, each unit dose contains 0.05 mg - 500 mg, preferably 0.5 mg - 200 mg, more preferably 0.1 mg - 100 mg of the compound of formula (I), enantiomer, racemate, pharmaceutically acceptable salt or a mixture thereof.

[0168] When the pharmaceutical composition contains an additional pharmaceutically active ingredient for treating or preventing cancer, the dosage of this active ingredient can generally be the conventional dosage in the prior art or lower.

[0169] The pharmaceutical composition of the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, etc., wherein the compound of formula (I) can be present in a suitable solid or liquid carrier or diluent. The pharmaceutical composition of the present invention can also be stored in a suitable sterilized device for injection or infusion. The pharmaceutical composition may also contain odorants, flavorants, etc.

[0170] The compound of formula (I) of the present invention or a pharmaceutical composition containing the compound of formula (I) can be clinically used in mammals (including humans) by administration routes such as oral, nasal, dermal, pulmonary or gastrointestinal routes. The preferred administration route is oral. The preferred daily dose is 0.5 mg - 200 mg / kg body weight, taken once or in divided doses. Regardless of the administration method, the optimal dose for an individual should be determined according to the specific treatment. Usually, it starts with a small dose and gradually increases the dose until the most suitable dose is found.

[0171] The effective dose of the active ingredient used may vary with the compound employed, the mode of administration, and the severity of the disease to be treated. However, generally satisfactory results are obtained when the compounds of the present invention are administered at a dose of about 1 - 300 mg / kg of animal body weight per day, preferably in 1 - 3 divided doses per day, or in a sustained-release form. 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 intimately mixed with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen may be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily depending upon the exigencies of the therapeutic situation, or the dose may be proportionally reduced.

[0172] The compounds or their pharmaceutically acceptable salts and compositions thereof may be administered orally as well as by intravenous, intramuscular or subcutaneous routes. From the standpoint of ease of preparation and administration, the preferred pharmaceutical compositions are solid compositions, especially tablets and solid-filled or liquid-filled capsules. Oral administration of the pharmaceutical compositions is preferred.

[0173] Solid carriers include: starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include: sterile water, polyethylene glycol, non-ionic surfactants and edible oils (such as corn oil, peanut oil and sesame oil), provided that they are suitable for the properties of the active ingredient and the particular mode of administration required. Adjuvants commonly used in the preparation of pharmaceutical compositions may also be advantageously included, such as flavoring agents, coloring agents, preservatives and antioxidants such as vitamin E, vitamin C, BHT and BHA.

[0174] The active compounds or their pharmaceutically acceptable salts and compositions thereof may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds (as the free base or pharmaceutically acceptable salts) may also be prepared in water suitably mixed with a surfactant (such as hydroxypropylcellulose). Dispersions may also be prepared in glycerol, liquid, polyethylene glycol and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparations contain preservatives to prevent the growth of microorganisms.

[0175] Dosage 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 to facilitate expulsion by syringe. They must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or a dispersion medium containing, for example, water, alcohols (such as glycerol, propylene glycol and liquid polyethylene glycol), their suitable mixtures and vegetable oils.

[0176] 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 the treatment or prevention of chronic pain diseases. When two or more drugs are administered in combination, generally, the effect is superior to that of the two drugs administered separately.

[0177] In a fourth aspect, the present invention provides the use of a polycyclic pyrimidine derivative as described in the first aspect, its pharmaceutically acceptable salt, its tautomer, its stereoisomer or the pharmaceutical composition as described in the third aspect alone or in combination with a MEK signaling pathway inhibitor in the preparation of a drug for the treatment of cancer or in the preparation of a SOS1 inhibitor; preferably, the cancer includes but is not limited to astrocytoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, pancreatic cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer, thyroid cancer, sarcoma, renal cancer and cholangiocarcinoma; more preferably, the cancer includes but is not limited to: pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer and cholangiocarcinoma. And / or the MEK signaling pathway inhibitor is such as a RAF, KRAS, MEK or ERK1 / 2 inhibitor.

[0178] Preferably, the dosage range of the polycyclic pyrimidine derivative as described in the first aspect, its pharmaceutically acceptable salt, its tautomer, its stereoisomer or the pharmaceutical composition as described in the third aspect is 10 - 100 mg / kg; and / or the dosage range of the MEK inhibitor is selected from 0.1 - 0.2 mg / kg; preferably, the dosage range of the polycyclic pyrimidine derivative, its pharmaceutically acceptable salt, its tautomer, its stereoisomer or the pharmaceutical composition is 25 - 50 mg / kg; and / or the dosage range of the MEK inhibitor is selected from 0.125 mg / kg; and / or the MEK inhibitor is Trametinib.

[0179] For the use as described in the fourth aspect, wherein the cancer is a RAS family-related cancer; preferably a KRAS, HRAS or NRAS G12C-related cancer, a KRAS, HRAS or NRAS G12D-related cancer, a KRAS, HRAS or NRAS G12S-related cancer, a KRAS, HRAS or NRAS G12A-related cancer, a KRAS, HRAS or NRAS G12V-related cancer, a KRAS, HRAS or NRAS G13D-related cancer, a KRAS, HRAS or NRAS G13C-related cancer, a KRAS, HRAS or NRAS Q61X-related cancer, a KRAS, HRAS or NRAS A146T-related cancer, a KRAS, HRAS or NRAS A146V-related cancer or a KRAS, HRAS or NRAS A146P-related cancer, etc.

[0180] On the other hand, there is provided a method for treating cancer, which comprises administering to a cancer patient a therapeutically effective amount of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the compound or its pharmaceutically acceptable salt. In one embodiment, the cancer is a RAS family-related cancer. In one embodiment, the cancer is a SOS-L-related cancer.

[0181] The compositions and methods provided by the present invention can be used for treating various cancers, including prostate, breast, brain, skin, cervical, testicular cancers, etc. More specifically, the cancers that can be treated by the compositions and methods of the present invention include, but are not limited to, tumor types such as astrocytoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, stomach, head and neck, hepatocellular carcinoma, laryngeal cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer, and thyroid cancer, and sarcomas.More specifically, these compounds can be used for the treatment of: Heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma; Lung: bronchial carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroma, mesothelioma; Gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid, VIPoma), small intestine (adenocarcinoma, lymphoma, carcinoid, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma, nephroblastoma (Wilms' tumor), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma); Liver: liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone: osteosarcoma (osteogenic sarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, juxtacortical osteosarcoma (parosteal osteosarcoma), benign tumors, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis), meninges (meningioma, meningeal sarcoma, glioma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pinealoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord fibroma, meningioma, glioma, sarcoma; Gynecology: uterus (endometrial carcinoma), cervix (cervical carcinoma, cervical dysplasia), ovary (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa theca cell tumor, Sertoli cell tumor, germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, embryonal rhabdomyosarcoma), fallopian tube (carcinoma); Hematology: blood (acute and chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndromes), Hodgkin's disease, non-Hodgkin lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus, lipoma, hemangioma, dermatofibroma, lupus erythematosus, psoriasis, and Adrenal gland: neuroblastoma.In certain embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL).

[0182] In one embodiment, the cancer is an 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 SOS1-related cancer. In certain embodiments, the SOS1-related cancer is lung adenocarcinoma, embryonal rhabdomyosarcoma, Sertoli cell testicular tumor, and cutaneous granular cell tumor.

[0183] Preferably, the cancer is pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer, or cholangiocarcinoma.

[0184] Term Explanation

[0185] Unless otherwise stated, some of the terms used in the present invention in the specification and claims are defined as follows:

[0186] As used herein, "KRAS G12C" refers to a mutant form of the mammalian KRAS protein that contains an amino acid in which glycine is replaced by cysteine at amino acid position 12.

[0187] The determination of the amino acid codons and residue positions of human KRAS is based on the amino acid sequence of UniProtKB / SwissProt P01116: variant p.Gly 12 Cys.

[0188] "KRAS G12D" refers to a mutant form of the mammalian KRAS protein that contains an amino acid in which aspartic acid replaces glycine at position 12. The amino acid codons and residue positions of human KRAS are determined according to the amino acid sequence determined by UniProtKB / Swi-protP01116: variant P.Gly 12 Asp.

[0189] "KRAS G12V" refers to a mutant form of the mammalian KRAS protein that contains an amino acid in which valine replaces glycine at position 12. The amino acid codons and residue positions of human KRAS are determined according to the amino acid sequence determined by UniProtKB / Swi-protP01116: variant P.Gly 12 Val.

[0190] "KRAS G12S" refers to a mutant form of a mammalian KRAS protein that contains an amino acid in which serine replaces glycine at amino acid position 12. The amino acid codons and residue positions of human KRAS are determined according to the amino acid sequence identified by UniProtKB / SwissProt P01116: variant p.Gly 12Ser.

[0191] As used herein, "KRAS G12A" refers to a mutant form of the mammalian KRAS protein that contains an amino acid substitution of alanine for glycine at amino acid position 12. The amino acid codons and residue positions of human KRAS are determined based on the amino acid sequence identified in UniProtKB / SwissProt P01116: Variant p.Gly 12Ala.

[0192] "KRAS G13D" refers to a mutant form of the mammalian KRAS protein that contains an amino acid substitution of aspartic acid for glycine at amino acid position 13. The amino acid codons and residue positions of human KRAS are determined based on the amino acid sequence determined by UniProtKB / SwissProt P01116: Variant p.gly 13Asp.

[0193] "KRAS G13C" refers to a mutant form of the mammalian KRAS protein that contains an amino acid substitution of cysteine for glycine at amino acid 13. The amino acid codons and residue positions of human KRAS are determined based on the amino acid sequence determined by UniProtKB / SwissProt-Prot P01116: Variant P.Gly 13Cys.

[0194] As used herein, "KRAS Q61L" refers to a mutant form of the mammalian KRAS protein that contains an amino acid substitution of leucine for glutamine at amino acid 61. The amino acid codons and residue positions of human KRAS are determined based on the amino acid sequence of UniProtKB / SwissProt P01116: Variant p.Gln61Leu.

[0195] As used herein, "KRAS A146T" refers to a mutant form of the mammalian KRAS protein that contains an amino acid substitution of threonine for alanine at amino acid position 146. The amino acid codons and residue positions of human KRAS are determined based on the amino acid sequence identified in UniProtKB / SwissProt P 01116: Variant p.al46Thr.

[0196] As used herein, "KRAS A146V" refers to a mutant form of the mammalian KRAS protein that contains an amino acid substitution of valine for alanine at amino acid position 146. The amino acid codons and residue positions of human KRAS are determined based on the amino acid sequence identified in UniProtKB / SwissProt P01116: Variant p.al46Val.

[0197] As used herein, "KRAS A146P" refers to a mutant form of the mammalian KRAS protein that contains a proline substitution for alanine at amino acid position 146. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProt P01116: variant p.A146Pro.

[0198] As used herein, "HRAS G12C" refers to a mutant form of the mammalian HRAS protein that contains a cysteine substitution for glycine at amino acid position 12. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence identified by UniProtKB / SwissProt P 01112: variant p.Gly 12 Cys.

[0199] "HRAS G12D" refers to a mutant of the mammalian HRAS protein that contains an aspartic acid substitution for glycine at amino acid position 12. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence of UniProtKB / SwissProt P01112: variant P.Gly 12Asp.

[0200] "HRAS G12S" refers to a mutant form of the mammalian HRAS protein that contains a serine substitution for glycine at position 12. The amino acid codons and residue positions of human HRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProtP01112: variant P.Gly 12Ser.

[0201] "HRAS G12A" refers to a mutant form of the mammalian HRAS protein that contains an alanine substitution for glycine at position 12. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProtP01112: variant P.Gly 12Ala.

[0202] "HRAS G13D" refers to a mutant form of the mammalian HRAS protein that contains an aspartic acid substitution for glycine at position 13. The amino acid codons and residue positions of human HRAS were determined based on the amino acid sequence identified by UniProtKB / Swi-protP01112: variant p.gly 13Asp.

[0203] "HRAS G13C" is a mutant of the mammalian HRAS protein, which contains an amino acid substitution of cysteine for glycine at the 13th amino acid. The amino acid codons and residue positions of human HRAS were determined based on the amino acid sequence of UniProtKB / SwissProt - Prot P01112: Variant P.Gly 13Cys.

[0204] As used herein, "HRAS Q61L" refers to a mutant form of the mammalian HRAS protein that contains an amino acid substitution of leucine for glutamine at amino acid position 61. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence identified by UniProtKB / SwissProt P01112 Variant p.Gln61Leu.

[0205] As used herein, "HRAS A146T" refers to a mutant form of the mammalian HRAS protein that contains an amino acid substitution of threonine for alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProt P01112: Variant p.A146Thr.

[0206] As used herein, "HRAS A146V" refers to a mutant form of the mammalian HRAS protein that contains an amino acid substitution of valine for alanine at amino acid position 146. One amino acid substitutes alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProt P 01112: Variant p.al46Val.

[0207] As used herein, "HRAS A146P" refers to a mutant form of the mammalian HRAS protein that contains an amino acid substitution of proline for alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified by UniProtKB / SwissProt P01112: Variant p.A146Pro.

[0208] "NRAS G12C" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of cysteine for glycine at amino acid 12. The amino acid codons and residue positions of human NRAS were determined based on UniProtKB / SwissProt - Prot P01111: Variant P.Gly 12Cys.

[0209] "NRAS G12D" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of aspartic acid for glycine at position 12. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / Swi-prot P01111: Variant P.Gly 12 Asp.

[0210] "NRAS G12S" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of serine for glycine at position 12. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P01111: Variant p.Gly 12Ser.

[0211] "NRAS G12A" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of alanine for glycine at position 12. The amino acid codons and residue positions of human KRAS were determined based on the amino acid sequence of UniProtKB / SwissProt P01111: Variant p.Gly 12Ala.

[0212] "NRAS G13D" as used herein refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of aspartic acid for glycine at amino acid position 13. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence determined by UniProtKB / SwissProt P 01111: Variant p.Gly 13Asp.

[0213] "HNRas G13C" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of cysteine for glycine at position 13. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence of UniProtKB / SwissProt P01111: Variant P.Gly 13Cys.

[0214] "HRAS Q61L" as used herein refers to a mutant form of a mammalian HRAS protein that contains an amino acid substitution of leucine for glutamine at amino acid position 61. The amino acid codons and residue positions of human HRAs were determined based on the amino acid sequence identified by UniProtKB / SwissProt P01112 Variant p.Gln61Leu.

[0215] As used herein, "NRAS A146T" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of threonine for alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P01111: Variant p.A146Thr.

[0216] As used herein, "NRAS A146V" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of valine for alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P01111: Variant p.A146Val.

[0217] As used herein, "NRAS A146P" refers to a mutant form of the mammalian NRAS protein that contains an amino acid substitution of proline for alanine at amino acid position 146. The amino acid codons and residue positions of human NRAS were determined based on the amino acid sequence identified in UniProtKB / SwissProt P01111: Variant p.A146Pro.

[0218] As used herein, "RAS family member" or "RAS family" refers to KRAS, HRAS, NRAS, and their activating mutants, including at positions G12, G13, Q61, and A146.

[0219] As used herein, 'RAS family-related 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 disorders are cancers associated with KRAS, HRAS, or NRAS G12C, cancers associated with KRAS, HRAS, or NRAS G12D, cancers associated with KRAS, HRAS, or NRAS G12V, cancers associated with KRAS, HRAS, or NRAS G12S, cancers associated with KRAS, HRAS, or NRAS G12A, cancers associated with KRAS, HRAS, or NRAS G13D, cancers associated with KRAS, HRAS, or NRAS G13C, cancers associated with KRAS, HRAS, or NRAS Q61L, cancers associated with KRAS, HRAS, or NRAS A146T, cancers associated with KRAS, HRAS, or NRAS A146V, or cancers associated with KRAS, HRAS, or NRAS 146P.

[0220] 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 SOS1 N233S and SOS1 N233Y mutations.

[0221] As used herein, "SOS1 N233S" refers to a mutant form of the mammalian SOS1 protein that contains an amino acid substitution in which serine replaces glutamine at amino acid position 233. The amino acid codons and residue positions of human SOS1 were determined based on the amino acid sequence identified by UniProtKB / SwissProt Q 07889: Variant p.Gln233Ser.

[0222] As used herein, "SOS1 N233Y" refers to a mutant form of the mammalian SOS1 protein that contains an amino acid substitution in which tyrosine replaces glutamine at amino acid position 233. The amino acid codons and residue positions of human SOS1 were determined based on the amino acid sequence identified by UniProtKB / SwissProt Q 07889: Variant p.Gln233Tyr.

[0223] As used herein, "SOS1 inhibitor" refers to the compounds of the present invention, as described herein, represented by formula (I). These compounds can negatively inhibit all or part of the interaction of SOS1 with RAS family mutants or SOS1 activating mutations, thereby reducing and / or regulating the nucleotide exchange activity of the RAS family member - SOS1 complex.

[0224] As used herein, "KRAS inhibitor" refers to inhibitors known in the art that target various KRAS mutations. Inhibitors can be targeted to, for example, KRAS G12C, KRAS G12D, KRAS G12V.

[0225] As used herein, "MEK signaling pathway" inhibitor refers to inhibitors known in the art that target RAF, MEK, and ERK1 / 2.

[0226] "NF-1 / NF-2 related disease or disorder" refers to a disease or disorder caused by a loss-of-function mutation in the neurofibromin (NF-1) gene or the neurofibromin 2 (NF-2) gene.

[0227] 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 frameshift mutation, including insertions and deletions, as well as homozygous deletion of the gene encoding the protein in the target cell or cancer cell, resulting in partial or complete loss of the presence, activity, and / or function of the encoded protein.

[0228] "Alkyl" refers to a saturated aliphatic hydrocarbon group, including a monovalent hydrocarbon group with 1 - 20 carbon atoms, or 1 - 10 carbon atoms, or 1 - 6 carbon atoms, or 1 - 4 carbon atoms, or 1 - 3 carbon atoms, or 1 - 2 carbon atoms, which can be a saturated straight-chain or branched-chain group. The alkyl group can be independently and optionally substituted by one or more substituents described in the present invention. Further examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group can be optionally substituted or unsubstituted.

[0229] "Alkenyl" refers to a monovalent hydrocarbon group with 2 - 12 carbon atoms, or 2 - 8 carbon atoms, or 2 - 6 carbon atoms, or 2 - 4 carbon atoms, which is a straight-chain or branched-chain group and has at least one C-C as an sp 2 double bond. The alkenyl group can be independently and optionally substituted by one or more substituents described in the present invention. Specific examples include, but are not limited to, vinyl, allyl, and butenyl, etc. The alkenyl group can be optionally substituted or unsubstituted.

[0230] "Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyl groups include spiro, fused, and bridged cycloalkyl groups. The cycloalkyl group can be optionally substituted or unsubstituted.

[0231] "Spiroalkyl" refers to a polycyclic group with 5 to 18 members, consisting of two or more cyclic structures, and sharing a single carbon atom (referred to as a spiro atom) between the monocyclic rings. The ring contains one or more double bonds, but no ring has a completely conjugated π-electron aromatic system. It is preferably 6 to 14 members, and more preferably 7 to 10 members. According to the number of spiro atoms shared between the rings, spiroalkyl groups are classified into monospiro, dispiro, or polyspiroalkyl groups, preferably monospiro and dispiroalkyl groups, preferably 4-member / 5-member, 4-member / 6-member, 5-member / 5-member, or 5-member / 6-member. Non-limiting examples of "spiroalkyl" include, but are not limited to:

[0232]

[0233] "Fused cycloalkyl" refers to a fully carbon polycyclic group having 5 to 18 ring atoms and containing two or more cyclic structures sharing a pair of carbon atoms with each other. One or more rings may contain one or more double bonds, but none of the rings has a fully conjugated π aromatic system of π electrons, preferably having 6 to 12 ring atoms, more preferably 7 to 10 ring atoms. 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 bicyclic alkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to:

[0234]

[0235] "Bridged cycloalkyl" refers to a fully carbon polycyclic group having 5 to 18 ring atoms and containing two or more cyclic structures sharing two non-directly connected carbon atoms with each other. One or more rings may contain one or more double bonds, but none of the rings has a fully conjugated aromatic system of π electrons, preferably having 6 to 12 ring atoms, more preferably 7 to 10 ring atoms. 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:

[0236]

[0237] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclic ring, wherein the ring connected to the parent structure is a cycloalkyl. Non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc.

[0238] "Heterocyclic group", "heterocycle" or "heterocyclic" are used interchangeably in this application and all refer to a saturated or partially unsaturated monocyclic, bicyclic or tricyclic non-aromatic heterocyclic group containing 3 - 12 ring atoms, wherein at least one ring atom is a heteroatom such as oxygen, nitrogen, sulfur atoms, etc. Preferably having a 5 - 7 membered monocyclic or 7 - 10 membered bi- or tricyclic ring, which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and / or sulfur. Examples of "heterocyclic group" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and piperazinyl. The heterocyclic ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group. The heterocyclic group may be optionally substituted or unsubstituted.

[0239] "Spiroheterocyclic group" refers to a polycyclic group with 5 to 18 members, two or more ring structures, and where the single rings share one atom with each other. The rings contain one or more double bonds, but none of the rings have a completely conjugated π-electron aromatic system. One or more of the ring atoms are selected from nitrogen, oxygen, sulfur, or S(O) m heteroatoms, and the remaining ring atoms are carbon, where m = 1 or 2. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. The spiroheterocyclic group is classified into monospiroheterocyclic group, dispiroheterocyclic group, or polyspiroheterocyclic group according to the number of shared spiro atoms, preferably monospiroheterocyclic group and dispiroheterocyclic group. More preferably, it is a 4-member / 4-member, 4-member / 5-member, 4-member / 6-member, 5-member / 5-member, or 5-member / 6-member monospiroheterocyclic group. Non-limiting examples of "spiroheterocyclic group" include, but are not limited to:

[0240]

[0241] "Fused heterocyclic group" refers to a fully carbon polycyclic group containing two or more ring structures sharing a pair of atoms. One or more of the rings may contain one or more double bonds, but none of the rings have a completely conjugated π-electron aromatic system. One or more of the ring atoms are selected from nitrogen, oxygen, sulfur, or S(O) m heteroatoms, and the remaining ring atoms are carbon, where m = 1 or 2. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. It can be classified into bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic groups according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-member / 5-member or 5-member / 6-member bicyclic fused heterocyclic groups. Non-limiting examples of "fused heterocyclic group" include, but are not limited to:

[0242]

[0243] "Bridged heterocyclic group" refers to a polycyclic group with 5 to 18 members, containing two or more ring structures, sharing two non-directly connected atoms with each other. One or more of the rings may contain one or more double bonds, but none of the rings have a completely conjugated π-electron aromatic system. One or more of the ring atoms are selected from nitrogen, oxygen, sulfur, or S(O) m heteroatoms, and the remaining ring atoms are carbon, where m = 1 or 2. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. It can be classified into bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic groups according to the number of constituent rings, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclic group" include, but are not limited to:

[0244]

[0245] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, where the rings can be connected in a fused manner. The term "aryl" includes aromatic groups such as phenyl, naphthyl, and tetrahydronaphthyl. Preferably, aryl is C6-C10 An aryl group, more preferably the aryl group is phenyl and naphthyl, and most preferably phenyl. The aryl group may be substituted or unsubstituted. The "aryl group" may be fused with a heteroaryl group, a heterocyclic group or a cycloalkyl group, wherein the aryl ring is connected to the parent structure. Non-limiting examples include, but are not limited to:

[0246]

[0247] "Heteroaryl group" 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 the "heteroaryl group" include, but are not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzodioxolyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolinyl, indazolyl, benzisothiazolyl, benzoxazolyl and benzisoxazolyl. The heteroaryl group may be optionally substituted or unsubstituted. The heteroaryl ring may be fused to an aryl group, a heterocyclic group or a cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. Non-limiting examples include, but are not limited to:

[0248]

[0249] "Alkoxy group" refers to a group of (alkyl-O-). Among them, the alkyl group is as defined herein. C1-C6 alkoxy groups are preferred. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, etc.

[0250] "Halogenated alkyl" refers to an alkyl group having one or more halogen substituents, wherein the alkyl group has the meaning as described in the present invention. Examples of the halogenated alkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, perfluoroethyl, 1,1-dichloroethyl, 1,2-dichloropropyl, etc.

[0251] "Hydroxyl group" refers to the -OH group.

[0252] "Halogen" refers to fluorine, chlorine, bromine and iodine, preferably fluorine, chlorine and bromine.

[0253] "Amino group" refers to -NH2.

[0254] "Cyano group" refers to -CN.

[0255] "Nitro group" refers to -NO2.

[0256] "Benzyl group" refers to -CH2-phenyl.

[0257] "Carboxyl" refers to -C(O)OH.

[0258] "Acetyl" refers to -C(O)CH3 or A c 。

[0259] "Carboxylate group" refers to -C(O)O(alkyl) or (cycloalkyl), where the definitions of alkyl and cycloalkyl are as described above.

[0260] "Optionally" means that the event described may but need not occur. For example, the statement "AR is optionally substituted by one or more R" c includes the cases where the AR group may or may not be substituted by one or more R. c c

[0261] "Substituted" means that one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 - 3 hydrogen atoms, are independently replaced by a corresponding number of substituents. It goes without saying that the substituents are only at their possible chemical positions, and those skilled in the art can determine (by experiment or theory) what substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with a free hydrogen may be unstable when bonded to a carbon atom with an unsaturated (such as olefinic) bond.

[0262] As used in this specification, "substituted" or "substitution", unless otherwise specified, means that the group may be substituted by one or more groups selected from the following: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic group, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate group, =O, -C(O)R b , -OC(O)R b , -NR b R b , -C(O)NR b R b , -NR b C(O)R b , -S(O)NR b R b or -S(O)2NR b R b , where the definition of R b is as described in general formula (I).

[0263] As used herein, the terms "subject", "individual", or "patient" are used interchangeably and refer to any animal, including mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and humans. In some embodiments, the patient is a human. In some embodiments, the subject has experienced and / or exhibits at least one symptom of a disease or disorder to be treated and / or prevented. In some embodiments, the subject has been determined or diagnosed with cancer having a KRAS G12 or G13 mutation (e.g., determined by a regulatory agency approved by the FDA, 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 (e.g., determined by a regulatory agency-approved assay or kit). The subject can be a cancer patient having a tumor that is positive for 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., determined by an approved regulatory agency, such as an FDA-approved assay or kit). The subject can be one 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 (e.g., the tumor is determined by an FDA-approved regulatory agency, kit, or assay). In some embodiments, the subject is suspected of having cancer associated with the KRAS G12 or G13 gene. In some embodiments, the subject has a clinical record indicating that the subject has a tumor having a KRAS G12C mutation (and optionally a clinical record indicating that the subject should be treated with any composition provided herein).

[0264] As used herein, the term "pediatric patient" refers to a patient who is less than 16 years of age at the time of diagnosis or treatment. The term "child" can also be divided into the following subcategories: neonate (from birth to the first month of life); infant (1 month to 2 years); child (2 years to 12 years); adolescent (12 years to 21 years (up to but not including the 22nd birthday)). Berhman RE, Kliegman R, Arvin AM, Nelson WE. Nelson Textbook of Pediatrics, 15th ed. Philadelphia: W.B. Saunders Co., 1996; Rudolph AM, et al. Rudolph's Pediatrics, 21st ed. New York: McGraw-Hill, 2002; and Avery MD, First LR. Pediatric Medicine, 2nd ed. Baltimore: Williams & Wilkins; 1994.

[0265] As used herein, an "effective amount" of a compound is an amount sufficient to negatively regulate or inhibit SOS1 enzyme activity.

[0266] As used herein, a "therapeutically effective dose" of a compound is an amount sufficient to ameliorate or in some way reduce symptoms, halt or reverse progression of a condition, or negatively regulate or inhibit SOS1 activity. Such a dose can be administered as a single dose or in accordance with a regimen so as to be effective.

[0267] As used herein, "treatment" means any manner of ameliorating or otherwise altering the symptoms or pathology of a patient's condition, disorder, or disease.

[0268] As described herein, "ameliorating the symptoms of a particular disease by use of a particular compound or pharmaceutical composition" means any reduction, whether permanent or temporary, durable or transitory, attributable to or associated with the use of the composition.

[0269] The definitions and conventions used herein for stereochemistry are generally referenced to the following:

[0270] S.P.Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. Compounds of the present invention may contain asymmetric or chiral centers and, therefore, exist in different stereoisomeric forms. 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, form part of the present invention. Diastereomers can be separated into the individual diastereomers on the basis of their physical and chemical differences by methods such as chromatography, crystallization, distillation, or sublimation. Enantiomers can be separated by converting the chiral isomeric mixture into a diastereomeric mixture, for example, by reaction with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers, and converting the individual diastereomers into the corresponding pure enantiomers. 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, i.e., 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 rotation of plane-polarized light by the compound, (-) or l indicating that the compound is levorotatory and the prefix (+) or d indicating that the compound is dextrorotatory. These stereoisomers have the same order of attachment of atoms or groups to each other but different three-dimensional structures. A particular stereoisomer may be an enantiomer, and a mixture of isomers is usually called a racemic mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereospecificity in a chemical reaction. The terms "racemic mixture" and "racemate" refer to a mixture of equimolar amounts of two enantiomers that lacks optical activity.

[0271] "Tautomers" or "tautomeric forms" refer to constitutional isomers of structures of different energies that can interconvert through a low energy barrier. For example, proton tautomers (i.e., prototropic tautomers) include interconversions through proton migration, such as the keto-enol and imine-enamine tautomerizations. Valence (oxidation state) tautomers include interconversions of the reorganized bonding electrons. Unless otherwise indicated, the structural formulas described in the present invention include all constitutional isomeric forms (such as enantiomers, diastereomers, and geometric isomers): for example, the R and S configurations containing an asymmetric center, the (Z) and (E) isomers of a double bond, and the (Z) and (E) conformational isomers. Therefore, the individual stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers, or geometric isomers are all within the scope of the present invention.

[0272] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention that are safe and effective when used in humans or animals. The salts of the compounds can be obtained by adding an appropriate amount of base or acid in a pure solution or a suitable inert solvent to obtain the corresponding addition salts. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, etc., and pharmaceutically acceptable acid addition salts include inorganic acid salts and organic acid salts. The inorganic acids and organic acids include hydrochloric acid, hydrobromic acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, acetic acid, maleic acid, malonic acid, succinic acid, fumaric acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, and methanesulfonic acid, etc. (see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977)).

[0273] The present invention provides an SOS1 inhibitor with a novel structure. The test results show that this polycyclic pyrimidine derivative exhibits excellent SOS1 inhibitory activity, and at the same time exhibits excellent safety and selectivity, and can be used to prepare drugs for treating diseases such as cancer, especially pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer, and cholangiocarcinoma. Description of the Drawings

[0274] Figure 1 It is a diagram showing the results of the effect of the compounds involved in the present invention on the KRAS / ERK1 / 2 signal transduction pathway in K-562 cells.

[0275] Figure 2 It is a diagram showing the in vivo antitumor effect of the SOS1 inhibitor involved in the present invention alone on the nude mouse model of KRAS G12D mutant PANC-1 pancreatic xenograft tumors.

[0276] Figure 3It is a graph showing the effect of the SOS1 inhibitor involved in the present invention alone on the body weight of nude mice in a PANC-1 pancreatic xenograft tumor nude mouse model with KRAS G12D mutation.

[0277] Figure 4 It is a graph showing the in vivo antitumor effect of the SOS1 inhibitor involved in the present invention alone or in combination with Trametinib on a nude mouse model of MIAPaCa-2 pancreatic xenograft tumor with KRAS G12C mutation.

[0278] Figure 5 It is a graph showing the effect of the SOS1 inhibitor involved in the present invention alone or in combination with Trametinib on the body weight of nude mice in a MIAPaCa-2 pancreatic xenograft tumor nude mouse model with KRAS G12C mutation. Detailed implementation manners

[0279] The method of the present invention will be described below through specific examples to make the technical solution of the present invention easier to understand and master, but the present invention is not limited thereto. In the following examples 1 The 1H NMR spectrum was measured using a Bruker instrument (400 MHz), and the chemical shift is expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. 1 Representation method of 1H NMR: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broadened, dd = doublet of doublets, dt = doublet of triplets. When coupling constants are provided, the unit is Hz.

[0280] The mass spectrum was obtained by measuring with an LC / MS instrument, and the ionization method was ESI.

[0281] Model of high performance liquid chromatograph: Agilent 1260, Thermo Fisher U3000; Model of chromatographic column: Waters xbrige C18 (4.6 * 150 mm, 3.5 μm); Mobile phase: A: ACN, B: Water (0.1% H3PO4); Flow rate: 1.0 mL / min; Gradient: 5% A for 1 min, increase to 20% A in 4 min, increase to 80% A within 8 min, 80% A for 2 min, back to 5% A within 0.1 min; Wavelength: 220 nm; Column oven: 35 °C.

[0282] For thin-layer chromatography silica gel plates, Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used. The specifications of the silica gel plates used in thin-layer chromatography (TLC) are 0.2 mm - 0.3 mm, and the specifications of the silica gel plates used for separating and purifying products by thin-layer chromatography are 0.4 mm - 0.5 mm.

[0283] For column chromatography, silica gel with 200 - 300 mesh of Yantai Huanghai silica gel is generally used as the carrier.

[0284] In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available raw materials and reagents are used directly without further purification. Unless otherwise specified, commercially available manufacturers include but are not limited to Sinopharm Group, J&K Scientific Ltd., Tokyo Chemical Industry Co., Ltd., Shanghai Bide Pharmatech Co., Ltd. and Shanghai Merck Chemical Technology Co., Ltd., etc.

[0285] CD3OD: deuterated methanol

[0286] CDCl3: deuterated chloroform

[0287] DMSO-d6: deuterated dimethyl sulfoxide

[0288] Pd2(db a )3: tris(dibenzylideneacetone)dipalladium

[0289] Pd(dppf)Cl2: [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium

[0290] XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

[0291] XPhos: 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl

[0292] HATU: 2-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate

[0293] TLC: thin-layer chromatography

[0294] HPLC: high performance liquid chromatography

[0295] purity: purity

[0296] &: and

[0297] The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1 L.

[0298] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0299] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20°C - 30°C.

[0300] The progress of the reaction in the examples was monitored by thin-layer chromatography (TLC). The eluent system for column chromatography used for purifying the compounds or the developing agent system for thin-layer chromatography included: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane:ethyl acetate. The volume ratio of the solvents varied according to the polarity of the compounds, and a small amount of acidic or basic reagents could also be added for adjustment, such as acetic acid or triethylamine.

[0301] Preparation of Intermediate

[0302] Intermediate 1

[0303] (R)-1-(3-Nitro-5-(trifluoromethyl)phenyl)ethanamine IN-1

[0304]

[0305] First Step 1-(3-Nitro-5-(trifluoromethyl)phenyl)ethan-1-one IN-1b

[0306] 3-Bromo-5-nitrobenzotrifluoride IN-1a (2.0 g, 7.41 mmol), tributyl(1-ethoxyvinyl)tin (3.5 g, 9.69 mmol) and Pd(PPh3)2Cl2 (520 mg, 0.74 mmol) were successively added to toluene (25 mL). Under nitrogen protection, the mixture was heated to 100°C and reacted overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, hydrochloric acid (15 mL, 3N) was added, and the mixture was stirred for 30 minutes. It was filtered through diatomaceous earth, 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 purified by silica gel column chromatography to obtain the title compound IN-1b as a yellow oil (1.25 g, yield 72%).

[0307] 1 H NMR (400 MHz, CDCl3) δ 8.94 (s, 1H), 8.68 (s, 1H), 8.53 (s, 1H), 2.75 (s, 3H).

[0308] Second Step (R,Z)-2-Methyl-N-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethylidene)propane-2-sulfinamide IN-1c

[0309] A mixture of compound IN-1b (1.25 g, 5.36 mmol), (R)-(+)-tert-butylsulfinamide (974 mg, 8.04 mmol) and tetraethyl titanate (10 mL, 47.70 mmol) was heated to 80 °C and reacted for 3 hours. TLC showed that a small amount of starting material remained. The reaction mixture 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 purified by silica gel column chromatography to obtain the title compound IN-1c as a yellow oil (1.01 g, yield 56%).

[0310] Step 3 (R)-2-Methyl-N-((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)propane-2-sulfinamide IN-1d

[0311] Compound IN-1c (260 mg, 0.77 mmol) was dissolved in tetrahydrofuran (2.5 mL) and water (0.05 mL). The temperature was lowered to -60 °C, and sodium borohydride (74 mg, 1.95 mmol) was added portionwise. After addition, the mixture was stirred at -60 °C for 1 hour. TLC showed that the reaction was complete. The reaction mixture was quenched by adding water dropwise, 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-1d as a white solid (150 mg, yield 58%).

[0312] 1 1H NMR (400 MHz, CDCl3) δ 8.43 - 8.42 (m, 2H), 7.95 (s, 1H), 4.75 - 4.69 (m, 1H), 3.55 (d, J = 4.4 Hz, 1H), 1.61 (d, J = 6.8 Hz, 3H), 1.25 (s, 9H).

[0313] Step 4 (R)-1-(3-Nitro-5-(trifluoromethyl)phenyl)ethanamine IN-1

[0314] Compound IN-1d (164 mg, 0.48 mmol) was dissolved in tetrahydrofuran (3 mL), and concentrated hydrochloric acid (0.5 mL) was added dropwise. After addition, the mixture was reacted at room temperature for 1 hour. TLC showed that the reaction was complete. The reaction mixture was adjusted to pH = 8 by adding saturated aqueous sodium carbonate solution dropwise, extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the title compound IN-1 as a yellow oil (105 mg, yield 92%).

[0315] LC-MS: m / z = 235.1 [M + H] +

[0316] Intermediate 2

[0317] (R)-3-(1-Aminoethyl)-5-(difluoromethyl)-4-fluoroaniline IN-2

[0318]

[0319] First step: 1-Bromo-3-(difluoromethyl)-2-fluorobenzene IN-2b

[0320] 2-Fluoro-3-bromobenzaldehyde IN-2a (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) was slowly added dropwise. After the addition was complete, the mixture was slowly warmed to room temperature and stirred for 1 hour. TLC showed that the raw material had completely reacted. The reaction solution was quenched by pouring it into saturated sodium bicarbonate aqueous solution, 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-2b as a yellow liquid (8.1 g, yield 73%).

[0321] 1 1H NMR (400 MHz, CDCl3) 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 = 54.8 Hz, 1H).

[0322] Second step: 1-(3-(Difluoromethyl)-2-fluorophenyl)ethanone IN-2c

[0323] Compound IN-2b (8.1 g, 36.00 mmol) was dissolved in 1,4-dioxane (80 mL). Triethylamine (9.1 g, 89.93 mmol) and tributyl(1-ethoxyvinyl)tin (15.6 g, 43.20 mmol) were successively added at room temperature. Nitrogen was bubbled for 15 minutes, Pd(PPh3)2Cl2 (250 mg, 0.36 mmol) was added, and nitrogen was replaced several times. The temperature was raised to 100 °C and stirred for 1 hour. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, dilute hydrochloric acid (14.4 mL, 72.0 mmol, 5 M) was added, and stirred at room temperature for 1 hour. TLC showed that the raw material had completely reacted. 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-2c as a yellow liquid (5.6 g, yield 83%).

[0324] 11H NMR (400 MHz, CDCl3) 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 = 54.8 Hz, 1H), 2.67 (d, J = 5.2 Hz, 3H).

[0325] Step 3 1-(3-(Difluoromethyl)-2-fluoro-5-nitrophenyl)ethan-1-one IN-2d

[0326] Potassium nitrate (45.6 g, 0.45 mmol) was dissolved in concentrated sulfuric acid (100 mL). The mixture was stirred at room temperature for 30 minutes, cooled to about 0 °C, and compound IN-2c (8.5 g, 45.18 mmol) was slowly added dropwise. The mixture was stirred at 0 °C for 5 minutes, and TLC showed that the reaction was complete. The reaction solution was slowly added to ice water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated aqueous sodium bicarbonate solution and saturated brine, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound IN-2d (9.0 g, yield 86%) as a pale yellow liquid.

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

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

[0329] Compound IN-2d (9.0 g, 38.60 mmol) was dispersed in tetraethyl orthotitanate (90 mL). (R)-(+)-tert-Butylsulfinamide (7.0 g, 57.76 mmol) was added at room temperature, and the mixture was slowly heated to 100 °C and stirred for 1 hour. TLC showed that the raw materials had completely reacted. The reaction solution was cooled to room temperature, poured into vigorously stirred ice water, and stirred for 10 minutes. The mixture was filtered, and the filter cake was repeatedly washed with ethyl acetate. The filtrate was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound IN-2e (9.0 g, crude product) as a yellow oil, which was directly used in the next step.

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

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

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

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

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

[0335] Compound IN-2f (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 exhausted. The reaction mixture was cooled to room temperature, poured into saturated aqueous sodium bicarbonate to adjust the pH to basic, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to give a brown liquid, which was allowed to solidify to give the title compound IN-2g as a brown solid (1.8 g, 95% yield).

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

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

[0338] Step 7 (R)-3-(1-aminoethyl)-5-(difluoromethyl)-4-fluoroaniline 2e

[0339] Compound IN-2g (300 mg, 1.28 mmol) was dissolved in ethanol and water (6 mL / 2 mL). Reductive iron powder (358 mg, 6.41 mmol) and ammonium chloride (346 mg, 6.69 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 2 hours. TLC detection showed that the reaction was complete. The reaction mixture was cooled to room temperature, filtered through diatomaceous earth, and the filtrate was concentrated to give the title compound IN-2 (350 mg, crude), which was directly used for the next step.

[0340] Intermediate 3

[0341] (R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethanamine IN-3

[0342]

[0343] First step (R,Z)-N-(1-(3-(Difluoromethyl)-2-fluorophenyl)ethylidene)-2-methylpropane-2-sulfinamide IN-3a

[0344] Compound IN-2c (5.6 g, 29.76 mmol) was dissolved in tetraethyl titanate (50 mL). At room temperature, (R)-(+)-tert-butylsulfinamide (10.8 g, 89.11 mmol) was added. The temperature was slowly raised to 100 °C and stirred for 1 hour. TLC showed that the raw materials had completely reacted. The reaction solution was cooled to room temperature and poured into vigorously stirred ice water, and stirred continuously for 10 minutes. It was filtered, and the filter cake was washed repeatedly with ethyl acetate. The filtrate was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow oily title compound IN-3a (9.5 g, crude product), which was directly used for the next step.

[0345] LC-MS: m / z = 292.1 [M+H] +

[0346] Second step (R)-N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide IN-3b

[0347] Compound IN-3a (9.5 g, crude product) was dissolved in tetrahydrofuran (100 mL) and water (2 mL). The temperature was lowered to about -60 °C, and sodium borohydride (3.7 g, 97.80 mmol) was added in batches. After addition, the temperature was slowly raised to room temperature and the reaction continued for 2 hours. TLC showed that the raw materials were exhausted. The reaction solution was poured into ice water for quenching, 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 yellow oily title compound IN-3b (2.9 g, overall yield of two steps 33%).

[0348] LC-MS: m / z = 294.1 [M+H] +

[0349] Third step (R)-1-(3-(Difluoromethyl)-2-fluorophenethylamine IN-3

[0350] Compound IN-3b (2.9 g, 9.88 mmol) was dissolved in tetrahydrofuran (50 mL). Concentrated hydrochloric acid (720 mg, 19.8 mmol, 12 N) was added at room temperature, and the mixture was heated to 80 °C and stirred for 1 h. TLC showed that the starting material was exhausted. The reaction mixture was cooled to room temperature, poured into saturated aqueous sodium bicarbonate 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 IN-3 (2.0 g, crude product), which was directly used in the next step.

[0351] LC-MS: m / z = 190.1 [M+H] +

[0352] 1 1H NMR (400 MHz, CDCl3) 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 = 55.2 Hz, 1H), 4.45 (q, J = 6.8 Hz, 1H), 1.80 (s, 2H), 1.43 (d, J = 6.4 Hz, 3H).

[0353] Intermediate 4

[0354] (S)-Ethyl 2-((tetrahydrofuran-3-yl)oxy)acetate IN-4

[0355]

[0356] First step (S)-Tetrahydrofuran-3-yl 4-nitrobenzoate IN-4b

[0357] Under nitrogen protection, (R)-(-)-3-Hydroxytetrahydrofuran IN-4a (10.0 g, 0.113 mol) was dissolved in tetrahydrofuran (130 mL). 4-Nitrobenzoic acid (18.9 g, 0.113 mol) and triphenylphosphine (35.6 g, 0.136 mol) were added at room temperature, and the temperature was cooled to 0 °C. A solution of diisopropyl azodicarboxylate (27.8 g, 0.137 mol) in tetrahydrofuran (20 mL) was added dropwise. After the addition was complete, the mixture was stirred at room temperature for 2 h. TLC showed that the starting material had reacted completely. The reaction mixture 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 give the title compound IN-4b (29.8 g, crude product, containing a small amount of triphenylphosphine oxide) as a white solid.

[0358] Second step (S)-3-Hydroxytetrahydrofuran IN-4c

[0359] Compound IN-4b (29.8 g, crude) was dissolved in ethanol (300 mL), and sodium carbonate (25.0 g, 0.236 mol) was added. The mixture was heated to 85 °C and reacted for 2 hours. TLC analysis showed that the raw materials had completely reacted. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography to obtain the oily title compound IN-4c (8.1 g, overall yield of two steps: 81%).

[0360] Step 3: (S)-tert-Butyl 2-((tetrahydrofuran-3-yl)oxy)acetate IN-4d

[0361] Compound IN-4c (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. tert-Butyl bromoacetate (9.9 g, 50.75 mmol) was added under stirring at room temperature. After addition, the mixture was stirred at 20 °C overnight. TLC analysis showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the yellow oily title compound IN-4d (5.5 g, yield: 80%).

[0362] 1 1H NMR (400 MHz, CDCl3) 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).

[0363] Step 4: (S)-2-((Tetrahydrofuran-3-yl)oxy)acetic acid IN-4e

[0364] Compound IN-4d (5.5 g, 27.19 mmol) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (30 mL) was added. The mixture was heated to 45 °C and reacted for 2 hours. TLC analysis showed that the reaction was basically complete. The reaction mixture was concentrated (using dichloromethane for azeotropic distillation three times) to obtain the pale yellow oily title compound IN-4e (13.5 g, crude), which was directly used in the next step.

[0365] Step 5: Ethyl (S)-2-((tetrahydrofuran-3-yl)oxy)acetate IN-4

[0366] Compound IN-4e (13.5 g, crude) was dissolved in ethanol (100 mL), and concentrated sulfuric acid (30 mL) was added at room temperature. The mixture was heated to 85 °C and reacted for 5 hours. The reaction mixture was cooled to room temperature, poured into ice-salt 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 slightly yellow oily title compound IN-4 (2.1 g, overall yield of two steps: 44%).

[0367] 1 1H NMR (400 MHz, CDCl3) 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 = 7.2 Hz, 3H).

[0368] Intermediate 5

[0369] tert-Butyl (R)-(3-(1-aminoethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-5

[0370]

[0371] First step: 3-Bromo-2-fluoro-5-nitrobenzoic acid IN-5b

[0372] Concentrated sulfuric acid (60 mL) was cooled to 0 °C, 2-Fluoro-5-nitrobenzoic acid IN-5a (10.0 g, 54.0 mmol) was added, and after dissolution, 1,3-dibromo-5,5-dimethylhydantoin (9.30 g, 32.4 mmol) was added. The mixture was warmed to room temperature and reacted overnight. The reaction was monitored by TLC and the starting material was completely reacted. The reaction mixture was poured into ice water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound IN-5b (15.0 g, crude), which was directly used in the next step.

[0373] Second step: (3-Bromo-2-fluoro-5-nitrophenyl)methanol IN-5c

[0374] Compound IN-5b (15.0 g, crude) was dissolved in tetrahydrofuran (100 mL). Under nitrogen protection, it was cooled to 0 °C, and borane-tetrahydrofuran solution (85.3 mL, 85.3 mmol, 10 M) was added dropwise. After addition, the mixture was warmed to room temperature and reacted overnight. The reaction was monitored by TLC and was complete. The reaction mixture was cooled to 0 °C, and methanol was added dropwise to quench until no gas was evolved. The mixture was concentrated, and the residue was dissolved in ethyl acetate, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the pale yellow solid title compound IN-5c (14.5 g, crude), which was directly used in the next step.

[0375] Third step: (5-Amino-3-bromo-2-fluorophenyl)methanol IN-5d

[0376] Compound IN-5c (14.5 g, crude) and stannous chloride (44.0 g, 232 mmol) were dissolved in ethyl acetate (200 mL). The solution was cooled to 0 °C, and concentrated hydrochloric acid (9.70 mL, 116 mmol, 12 M) was added dropwise. After addition, the mixture was allowed to warm to room temperature and stirred for 3 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was quenched with ice water, and the pH was adjusted to 14 with aqueous sodium hydroxide solution (2 N). The mixture was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound IN-5d as a pale yellow solid (13.5 g, crude), which was used directly in the next step.

[0377] LC-MS: m / z = 220.0 [M+H] +

[0378] Step 4 tert-Butyl (3-bromo-4-fluoro-5-(hydroxymethyl)phenyl)carbamate IN-5e

[0379] Compound IN-5d (12.8 g, crude) was dissolved in N,N-dimethylformamide (100 mL). Di-tert-butyl dicarbonate (25.3 g, 116 mmol) was added at room temperature, and the mixture was heated to 60 °C and stirred for 3 hours. The reaction was monitored by TLC and the starting material was found to have reacted completely. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound IN-5e as a pale yellow solid (18.6 g, crude), which was used directly in the next step.

[0380] LC-MS: m / z = 318.0 [M-H] -

[0381] Step 5 tert-Butyl (3-bromo-4-fluoro-5-formylphenyl)carbamate IN-5f

[0382] Compound IN-5e (18.6 g, crude) was dissolved in dichloromethane (200 mL). The solution was cooled to 0 °C, and Dess-Martin periodinane (36.9 g, 87.0 mmol) was added. The reaction was allowed to proceed at room temperature for 2 hours. The reaction was monitored by TLC and the starting material was found to have reacted completely. The reaction mixture was cooled to 0 °C, quenched with saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound IN-5f as a yellow solid (18.4 g, crude), which was used directly in the next step.

[0383] Step 6 tert-Butyl (3-bromo-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-5g

[0384] Compound IN-5f (18.4 g, crude) was dissolved in dichloromethane (200 mL), cooled to 0 °C, and diethylaminosulfur trifluoride (19.6 g, 122 mmol) was added dropwise. After addition, the reaction was continued at 0 °C for 1 hour, then raised to room temperature and reacted for 2 hours. TLC detected that the reaction was complete. The reaction solution was quenched with water, extracted with dichloromethane, 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 yellow solid titled compound IN-5g (19.2 g, crude), which was directly used for the next reaction.

[0385] Step 7 tert-Butyl (3-acetyl-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-5h

[0386] Compound IN-5g (19.2 g, crude), tributyl(1-ethoxyvinyl)tin (20.1 g, 56.5 mmol) and bis(triphenylphosphine)palladium(II) dichloride (793 mg, 1.13 mmol) were successively added to toluene (100 mL). Under nitrogen protection, the temperature was raised to 100 °C and stirred overnight. TLC detected that the raw materials disappeared. The reaction solution was cooled to room temperature, dilute hydrochloric acid (50 mL, 2N) was added and stirred for 3 hours. TLC detected that the reaction was complete. The reaction solution was added with 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 yellow solid titled compound IN-5h (9.1 g, 52% yield in seven steps).

[0387] Step 8 tert-Butyl (R,Z)-(3-(1-((tert-butylsulfinyl)imino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-5i

[0388] Compound IN-5h (5.50 g, 18.1 mmol) and tert-butylsulfinamide (3.30 g, 27.2 mmol) were dissolved in tetrahydrofuran (80 mL). Isopropyl titanate (10.3 g, 36.3 mmol) was added at room temperature, and the temperature was raised to 80 °C and stirred overnight. TLC detected that the reaction was complete. The reaction solution was cooled to room temperature, quenched with water, the insoluble substances were filtered off, the filtrate was 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 yellow solid titled compound IN-5i (4.60 g, 62% yield).

[0389] Step 9 tert-Butyl (3-((R)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-5j

[0390] Compound IN-5i (4.60 g, 11.3 mmol) was dissolved in tetrahydrofuran (50 mL) and water (3 mL). The temperature was lowered to -20 °C, and sodium borohydride (856 mg, 22.6 mmol) was added. The mixture was allowed to warm to room temperature and stirred for 1 hour. The reaction was monitored by TLC and was found to be complete. The reaction mixture was quenched with 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-5j as a yellow oil (1.92 g, yield 42%).

[0391] Step 10 (R)-(tert-butyl 3-(1-aminoethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-5

[0392] Compound IN-5j (1.92 g, 4.70 mmol) was dissolved in tetrahydrofuran (20 mL). Concentrated hydrochloric acid (0.80 mL, 9.40 mmol, 12 N) was added, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC and was found to be complete. The reaction mixture was quenched with water, neutralized with saturated aqueous sodium bicarbonate, 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-5 as a yellow oil (1.41 g, yield 99%).

[0393] LC-MS: m / z = 305.1 [M+H] +

[0394] 1 1H NMR (400 MHz, CDCl3) δ 7.64 - 7.57 (m, 1H), 7.54 - 7.46 (m, 1H), 6.83 (t, J = 54.8 Hz, 1H), 6.80 (br, 1H), 4.43 (q, J = 6.8 Hz, 1H), 2.51 (s, 2H), 1.50 (s, 9H), 1.43 (d, J = 6.8 Hz, 3H).

[0395] Intermediate 6

[0396] (R)-(isopropyl 3-(1-aminoethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-6

[0397]

[0398] Step 1 1-(5-amino-3-(difluoromethyl)-2-fluorophenyl)ethan-1-one IN-6a

[0399] Compound IN-5h (3.0 g, 9.89 mmol) was dissolved in dichloromethane (30 mL), trifluoroacetic acid (10 mL) was added, and the reaction was carried out at room temperature for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was concentrated, the residue was neutralized with saturated aqueous sodium carbonate solution, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow oily title compound IN-6a (2.5 g, crude product), which was directly used in the next step.

[0400] Second step: Benzyl (3-acetyl-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-6b

[0401] Compound IN-6a (2.5 g, crude product) was dissolved in tetrahydrofuran (20 mL) and water (8 mL), sodium carbonate (5.2 g, 49.06 mmol) was added, benzyl chloroformate (2.0 g, 11.72 mmol) was added dropwise. After the addition, the reaction was carried out at room temperature for 4 hours. The reaction was monitored by TLC and found to be basically complete. Water was added to the reaction solution, and it was 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 yellow solid title compound IN-6b (2.2 g, overall yield of two steps 67%).

[0402] LC-MS: m / z = 393.1 [M+H] +

[0403] Third step: Isopropyl (R,Z)-(3-(1-((tert-butylsulfinyl)imino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-6c

[0404] Compound IN-6b (2.2 g, 6.52 mmol) and (R)-(+)-tert-butylsulfinamide (870 mg, 7.18 mmol) were dissolved in tetrahydrofuran (20 mL). Tetraisopropyl titanate (9.2 g, 32.37 mmol) was added at room temperature, and the mixture was heated to 70 °C and reacted overnight. The reaction was monitored by TLC and found to be basically complete. The reaction solution was cooled to room temperature, ethyl acetate and water were added, and the mixture was stirred vigorously for half an hour, then filtered. The filtrate was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the pale yellow solid title compound IN-6c (1.4 g, yield 55%).

[0405] Fourth step: Isopropyl (3-((R)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate IN-6d

[0406] Compound IN-6c (1.4 g, 3.57 mmol) was dispersed in tetrahydrofuran (15 mL) and water (1.5 mL). The temperature was lowered to -60 °C, and sodium borohydride (675 mg, 17.85 mmol) was added in portions. After addition, the mixture was slowly warmed to room temperature and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was poured into ice water to quench the reaction, 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-6d as a pale yellow solid (710 mg, yield 50%).

[0407] Step 5 (R)-(3-(1-Aminoethyl)-5-(difluoromethyl)-4-fluorophenyl) isopropyl carbamate IN-6

[0408] Compound IN-6d (710 mg, 1.80 mmol) was dissolved in tetrahydrofuran (10 mL), concentrated hydrochloric acid (0.2 mL, 2.36 mmol, 12 N) was added, and the mixture was reacted at room temperature for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was neutralized with saturated aqueous sodium carbonate 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-6 as a white solid (495 mg, yield 95%).

[0409] LC-MS: m / z = 291.1 [M+H] +

[0410] 1 H NMR (400 MHz, CDCl3) δ 7.68 - 7.53 (m, 2H), 7.41 - 7.30 (m, 1H), 6.83 (t, J = 54.8 Hz, 1H), 5.03 - 4.93 (m, 1H), 4.47 (q, J = 6.4 Hz, 1H), 3.59 (br, 2H), 1.45 (d, J = 6.8 Hz, 3H), 1.28 (d, J = 6.4 Hz, 6H).

[0411] Example 1

[0412] N 4 -((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-N 6 -((S)-tetrahydrofuran-3-yl)quinazoline-4,6-diamine 1

[0413]

[0414] Step 1 5-Hydroxy-4-methoxy-2-nitrobenzoic acid 1b

[0415] 2-Nitro-4,5-dimethoxybenzoic acid 1a (15.0 g, 66.03 mmol) was dissolved in an aqueous sodium hydroxide solution (60 mL, 6 M). The reaction mixture was heated to 100 °C and stirred for 3 hours. TLC analysis showed that the starting material had completely reacted. The reaction mixture was cooled to room temperature, and the pH was adjusted to 2 - 3 with hydrochloric acid. The mixture was extracted with ethyl acetate, washed with brine, and the organic phase was dried and concentrated to obtain the yellow solid title compound 1b (14.1 g, crude), which was directly used in the next step.

[0416] 1 1H NMR (400 MHz, DMSO-d6) δ 13.44 (s, 1H), 10.76 (s, 1H), 7.56 (s, 1H), 7.08 (s, 1H), 3.90 (s, 3H).

[0417] The second step: Methyl 5-hydroxy-4-methoxy-2-nitrobenzoate 1c

[0418] Compound 1b (14.1 g, crude) was dissolved in methanol (150 mL). Thionyl chloride (23.6 g, 198.37 mmol) was added dropwise. The reaction mixture was heated to 80 °C and stirred for 28 hours. TLC analysis showed that the reaction was complete. The reaction mixture was cooled to room temperature, concentrated, and the residue was dissolved in ethyl acetate, washed with water and saturated brine. The organic phase was dried and concentrated to obtain the yellow solid title compound 1c (14.1 g, crude), which was directly used in the next step.

[0419] The third step: Methyl 5-(benzyloxy)-4-methoxy-2-nitrobenzoate 1d

[0420] Compound 1c (14.1 g, crude) and benzyl bromide (12.7 g, 74.26 mmol) were dissolved in dimethyl sulfoxide (100 mL). Cesium carbonate (40.4 g, 123.99 mmol) was added, and the mixture was stirred at room temperature overnight. TLC analysis showed that the reaction was complete. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with brine, and the organic phase was dried and concentrated to obtain the yellow oily title compound 1d (18.4 g, crude), which was directly used in the next step.

[0421] The fourth step: Methyl 2-amino-5-(benzyloxy)-4-methoxybenzoate 1e

[0422] Compound 1d (18.4 g, crude) was dissolved in ethanol (200 mL) and water (40 mL). Iron powder (16.2 g, 0.29 mol) and ammonium chloride (15.5 g, 0.29 mol) were added. The reaction mixture was heated to 90 °C and stirred for 2 hours. TLC analysis showed that the reaction was complete. The reaction mixture was filtered through diatomaceous earth while hot. The filtrate was concentrated, and the residue was dissolved in ethyl acetate, washed with brine, and the organic phase was dried and concentrated. The residue was purified by silica gel column chromatography to obtain the yellow oily title compound 1e (14.6 g, overall yield of four steps 77%).

[0423] Step 5 6-(Benzyloxy)-7-methoxy-2-methylquinazolin-4(3H)-one 1f

[0424] Dissolve compound 1e (1.0 g, 3.48 mmol) in acetonitrile (10 mL), add 4 M hydrochloric acid / 1,4-dioxane solution (20 mL, 4 M), and heat the reaction mixture to 90 °C and stir overnight. Cool the reaction mixture to room temperature, filter the precipitated solid, and collect the solid. Dissolve the solid in water, adjust the pH to 8 with saturated aqueous sodium bicarbonate solution, filter to collect the solid, and dry the solid to obtain the yellow solid title compound 1f (930 mg, crude), which is directly used in the next step.

[0425] LC-MS: m / z = 297.1 [M+H] +

[0426] Step 6 6-(Benzyloxy)-4-chloro-7-methoxy-2-methylquinazoline 1g

[0427] Dissolve compound 1f (930 mg, crude) in phosphorus oxychloride (20 mL), add N,N-diisopropylethylamine (1.2 g, 9.29 mmol), heat the reaction mixture to 100 °C and stir for 8 hours, and monitor the reaction by TLC until completion. Cool the reaction mixture to room temperature, concentrate it, dissolve the residue in dichloromethane, and quench it by dropping it into a cooled aqueous sodium bicarbonate solution. Wash with brine, concentrate the organic phase, and purify it by column chromatography to obtain the brown solid title compound 1g (400 mg, 36% yield over two steps).

[0428] LC-MS: m / z = 315.1 [M+H] +

[0429] Step 7 (R)-6-(Benzyloxy)-7-methoxy-2-methyl-N-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)quinazolin-4-amine 1h

[0430] Dissolve compound 1g (400 mg, 1.27 mmol) and compound IN-1 (328 mg, 1.40 mmol) in dimethyl sulfoxide (10 mL), add N,N-diisopropylethylamine (493 mg, 3.81 mmol), heat the reaction mixture to 80 °C and stir overnight, and monitor the reaction by TLC until completion. Cool the reaction mixture to room temperature, add water, extract with ethyl acetate, wash with brine, dry the organic phase, and concentrate to obtain the yellow solid title compound 1h (700 mg, crude), which is directly used in the next step.

[0431] Step 8 (R)-7-Methoxy-2-methyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)quinazolin-6-ol 1i

[0432] Compound 1h (700 mg, crude) was dissolved in trifluoroacetic acid (6 mL). The reaction mixture was heated to 80 °C and stirred for 2 h. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature and concentrated. The residue was dissolved in dichloromethane, washed with water, and the organic phase was dried and concentrated to give the yellow solid title compound 1i (577 mg, crude), which was used directly in the next step.

[0433] LC-MS: m / z = 423.2 [M+H] +

[0434] Step 9 (R)-7-Methoxy-2-methyl-4-((1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)quinazolin-6-yl trifluoromethanesulfonate 1j

[0435] Compound 1i (577 mg, crude) was dissolved in dichloromethane (20 mL). Pyridine (216 mg, 2.73 mmol) and trifluoromethanesulfonic anhydride (578 mg, 2.05 mmol) were added. The reaction mixture was stirred at room temperature for 3 h. The reaction was monitored by TLC and was found to be complete. The reaction mixture was diluted with water and extracted with dichloromethane, washed with brine, and the organic phase was concentrated and purified by silica gel column chromatography to give the yellow oil title compound 1j (460 mg, 65% yield over three steps).

[0436] LC-MS: m / z = 555.2 [M+H] +

[0437] Step 10 7-Methoxy-2-methyl-N 4 -((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)-N 6 -((S)-tetrahydrofuran-3-yl)quinazoline-4,6-diamine 5l

[0438] Compound 1j (130 mg, 0.23 mmol) and (S)-3-aminotetrahydrofuran 1k (61 mg, 0.70 mmol) were dissolved in toluene (20 mL). Pd2(dba)3 (43 mg, 0.047 mmol), BINAP (59 mg, 0.095 mmol) and cesium carbonate (152 mg, 0.47 mmol) were added. The reaction mixture was heated to 100 °C and stirred for 5 h under nitrogen protection. The reaction was monitored by TLC and the starting materials were found to have reacted almost completely. The reaction mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate, washed with brine. The organic phase was dried and concentrated. The residue was purified by silica gel column chromatography to give the yellow solid title compound 1l (56 mg, 49% yield).

[0439] LC-MS: m / z = 492.2 [M+H] +

[0440] The eleventh step N 4 -((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-N 6 -((S)-tetrahydrofuran-3-yl)quinazoline-4,6-diamine 1

[0441] Compound 1k (56 mg, 0.11 mmol) was dissolved in ethanol (6 mL) and water (1 mL). Iron powder (32 mg, 0.57 mmol) and ammonium chloride (30 mg, 0.56 mmol) were added. The reaction mixture was heated to 90 °C and stirred for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was filtered through diatomaceous earth while hot, and the filtrate was concentrated. The residue was purified by Prep-TLC to give the title compound 1 as a pale yellow solid (26 mg, yield 49%).

[0442] LC-MS: m / z = 462.3 [M+H] +

[0443] 1 1H NMR (400 MHz, CD3OD) δ 7.20 (s, 1H), 6.98 (s, 1H), 6.97 (s, 1H), 6.94 (s, 1H), 6.80 (s, 1H), 5.66 (q, J = 7.2 Hz, 1H), 4.35 - 4.29 (m, 1H), 4.07 - 3.96 (m, 5H), 3.91 - 3.85 (m, 1H), 3.75 - 3.72 (m, 1H), 2.47 (s, 3H), 2.44 - 2.35 (m, 1H), 1.99 - 1.92 (m, 1H), 1.65 (d, J = 7.2 Hz, 3H). (96.49% purity by HPLC)

[0444] Example 2

[0445] N-((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-4-amine 2

[0446]

[0447] The first step 6-chloro-2-methyl-4-pyrimidinamine 2b

[0448] 4,6-Dichloro-2-methylpyrimidine 2a (15.2 g, 93.25 mmol) was added to aqueous ammonia solution (150 mL, 25%). The reaction mixture was heated to 65 °C and reacted for 3 hours. TLC showed that the raw material was completely reacted. The reaction mixture was concentrated to obtain the title compound 2b as a white solid (11.9 g, crude product), which was directly used in the next step.

[0449] LC-MS: m / z = 144.1 [M+H] +

[0450] The second step: 6-chloro-5-iodo-2-methyl-4-pyrimidinamine 2c

[0451] Compound 2b (11.9 g, crude) was added to acetic acid (120 mL), and N-iodosuccinimide (18.64 g, 82.85 mmol) was added portionwise at room temperature. The mixture was heated to 70 °C and reacted for 4 hours. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to room temperature, adjusted to neutral with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, 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 the white solid title compound 2c (11.82 g, crude), which was directly used for the next step.

[0452] LC-MS: m / z = 270.0 [M+H] +

[0453] The third step: 6-chloro-2-methyl-5-vinyl-4-pyrimidinamine 2d

[0454] Compound 6c (18.0 g, crude) was dissolved in ethylene glycol dimethyl ether (180 mL). At room temperature, potassium vinyltrifluoroborate (17.89 g, 0.13 mol), potassium phosphate (35.9 g, 0.17 mol), and Pd(dppf)Cl2 dichloromethane complex (1.64 g, 2.01 mmol) were added in sequence. The mixture was purged with nitrogen three times, heated to 80 °C and reacted for 3 hours. TLC showed that the raw materials had reacted completely. 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 white solid title compound 2d (6.1 g, overall yield of three steps 25%).

[0455] The fourth step: 4-amino-6-chloro-2-methylpyrimidine-5-carbaldehyde 2e

[0456] Compound 2d (5.00 g, 29.48 mmol) was dissolved in acetone and water (120 mL / 30 mL), sodium periodate (14.32 g, 66.95 mmol) and an aqueous solution of potassium osmate (catalytic amount) were added, and the reaction was carried out at room temperature for 2 hours. TLC showed that the raw materials had reacted completely. 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 off-white solid title compound 2e (2.5 g, yield 49%).

[0457] LC-MS: m / z = 172.1 [M+H] +

[0458] Step 5 (S)-4-Chloro-2-methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-7(8H)-one 2f

[0459] Under nitrogen protection, compound 2e (2.50 g, 14.57 mmol) was dissolved in tetrahydrofuran (500 mL). Intermediate IN-4 (3.30 g, 18.94 mmol) was added at room temperature, and the mixture was cooled to -60 °C. Lithium diisopropylamide (22 mL, 43.86 mmol, 2.0 M) was added dropwise. After the addition, the reaction was carried out at -60 °C for 1 hour and then slowly warmed to room temperature and reacted overnight. The reaction solution was quenched with 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 2f as a pale yellow solid (1.0 g, yield 24%).

[0460] LC-MS: m / z = 282.1 [M+H] +

[0461] Step 6 (S)-4-Chloro-7-methoxy-2-methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidine 2g

[0462] Compound 2f (70 mg, 0.25 mmol) was dissolved in chloroform (5 mL). Silver carbonate (102 mg, 0.37 mmol) and methyl iodide (705 mg, 4.97 mmol) were added at room temperature, and the mixture was heated to 65 °C and reacted overnight. TLC showed that some starting materials remained. The reaction solution was cooled to room temperature, filtered, the filter cake was washed with dichloromethane, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 2g as a white solid (56 mg, yield 77%).

[0463] LC-MS: m / z = 296.1 [M+H] +

[0464] 1 1H NMR (400 MHz, CDCl3) δ 7.34 (s, 1H), 5.06 - 5.01 (m, 1H), 4.17 (s, 3H), 4.17 - 3.97 (m, 3H), 3.93 - 3.87 (m, 1H), 2.77 (s, 3H), 2.37 - 2.26 (m, 1H), 2.23 - 2.15 (m, 1H).

[0465] Step 7 7-Methoxy-2-methyl-N-((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-4-amine 2h

[0466] Compound 2g (44 mg, 0.15 mmol) was dissolved in dimethyl sulfoxide (3 mL). At room temperature, N,N-diisopropylethylamine (58 mg, 0.45 mmol) and compound IN-1 (38 mg, 0.16 mmol) were added. The mixture was heated to 100 °C and reacted for 10 hours. TLC showed that the raw materials were completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid title compound 2h (55 mg, yield 75%).

[0467] LC-MS: m / z = 494.2 [M+H] +

[0468] Step 8 N-((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[2,3-d]pyrimidin-4-amine 2

[0469] Compound 2h (55 mg, 0.11 mmol) was dissolved in ethanol (10 mL) and water (3 mL). At room temperature, iron powder (31 mg, 0.56 mmol) and ammonium chloride (28 mg, 0.52 mmol) were added. The mixture was heated to 90 °C and reacted for 3 hours. TLC showed that the raw materials were completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the white solid title compound 2 (35 mg, yield 67%).

[0470] LC-MS: m / z = 464.3 [M+H] +

[0471] 1 1H NMR (400 MHz, CD3OD) δ 8.05 (s, 1H), 7.01 - 6.95 (m, 2H), 6.81 (s, 1H), 5.68 (q, J = 7.2 Hz, 1H), 5.20 - 5.15 (m, 1H), 4.11 (s, 3H), 4.06 - 3.95 (m, 3H), 3.94 - 3.90 (m, 1H), 2.55 (s, 3H), 2.41 - 2.31 (m, 1H), 2.23 - 2.16 (m, 1H), 1.68 (d, J = 7.2 Hz, 3H). (99.74% purity by HPLC)

[0472] Example 3

[0473] N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[3,4-d]pyrimidin-4-amine 3

[0474]

[0475] The first step: 2-chloro-5-nitroisonicotinic acid 3b

[0476] 2-Chloro-5-nitro-4-methylpyridine 3a (20 g, 115.9 mmol) was dissolved in concentrated sulfuric acid (150 mL), and chromium trioxide (28.97 g, 289.7 mmol) was added portionwise at room temperature. The reaction was carried out overnight at room temperature, and TLC showed that the reaction was complete. The reaction solution was poured into ice water, stirred for 1 hour, filtered, and the filter cake was washed with water and dried to obtain the solid off-white solid title compound 3b (21.6 g, crude product), which was directly used for the next step.

[0477] The second step: 2-chloro-5-nitroisonicotinamide 3c

[0478] Compound 3b (10.0 g, 49.37 mmol) was heated under reflux in thionyl chloride (100 mL) for 3 hours, and TLC detected that the reaction was complete. The reaction solution was cooled to room temperature, concentrated, and the residue was dissolved in tetrahydrofuran (50 mL). It was slowly added dropwise to ammonia water (100 mL) cooled to about 0 °C. After addition, the reaction was carried out at 0 °C for 0.5 hour, and TLC detected that the reaction was complete. Water was added to the reaction solution, and it was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 3c (8.5 g, crude product), which was directly used for the next step.

[0479] The third step: (S)-5-nitro-2-((tetrahydrofuran-3-yl)oxy)isonicotinamide 3d

[0480] Compound 3c (3.0 g, crude product) was dissolved in acetonitrile (30 mL), cesium carbonate (9.69 g, 29.74 mmol) and S-3-hydroxytetrahydrofuran IN-4c (3.27 g, 37.11 mmol) were added at room temperature, and the temperature was raised to 30 °C and the reaction was carried out overnight. TLC showed that a small amount of raw material was not completely reacted. Water was added to the reaction solution, and it 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 yellow solid title compound 3d (1.67 g, overall yield of three steps 35%).

[0481] LC-MS: m / z = 254.1 [M+H] +

[0482] The fourth step: (S)-5-amino-2-((tetrahydrofuran-3-yl)oxy)isonicotinamide 3e

[0483] Compound 3d (1.67 g, 6.60 mmol) was dissolved in ethanol (30 mL) and water (10 mL). Iron powder (1.84 g, 32.95 mmol) and ammonium chloride (1.69 g, 31.59 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 3 hours. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, the filter cake was washed with ethanol, the filtrate was concentrated, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid titled compound 3e (1.38 g, crude product), which was directly used for the next step.

[0484] LC-MS: m / z = 224.2 [M+H] +

[0485] Step 5 (S)-2-Methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[3,4-d]pyrimidin-4-ol 3f

[0486] Compound 3e (300 mg, crude product) was dissolved in triethyl orthoacetate (6 mL), and the temperature was raised to 120 °C and reacted for 20 hours. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to room temperature, filtered, the filter cake was washed with methyl tert-butyl ether, and dried to obtain the white solid titled compound 3f (170 mg, crude product), which was directly used for the next step.

[0487] LC-MS: m / z = 248.1 [M+H] +

[0488] Step 6 (S)-4-Chloro-2-methyl-6-((tetrahydrofuran-3-yl)oxy)pyrido[3,4-d]pyrimidine 3g

[0489] Compound 3f (170 mg, crude product) was suspended in 1,4-dioxane (10 mL). N,N-Diisopropylethylamine (355 mg, 2.75 mmol) and phosphorus oxychloride (421 mg, 2.75 mmol) were added at room temperature, and the mixture was heated to 100 °C and reacted for 4 hours. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to 0 °C, quenched 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, concentrated, and the crude product was purified by silica gel column chromatography to obtain the pale yellow solid titled compound 3g (150 mg, overall yield in three steps 29%).

[0490] Step 7 N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)pyrido[3,4-d]pyrimidin-4-amine 3

[0491] Compound 3g (65 mg, 0.26 mmol) was dissolved in dimethyl sulfoxide (3 mL). At room temperature, N,N-diisopropylethylamine (100 mg, 0.77 mmol) and intermediate IN-2 (53 mg, 0.26 mmol) were added. The mixture was heated to 80 °C and reacted for 1.5 h. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain a yellow solid, and then further purified by Prep-HPLC to obtain a pale yellow solid, the title compound 3 (21 mg, yield 19%).

[0492] LC-MS: m / z = 434.2 [M+H] +

[0493] 1 1H NMR (400 MHz, CD3OD) δ 8.66 (s, 1H), 7.58 (s, 1H), 7.02 - 6.81 (m, 2H), 6.78 - 5.68 (m, 1H), 5.73 (q, J = 7.2 Hz, 1H), 5.65 - 5.657 (m, 1H), 4.08 - 3.87 (m, 4H), 2.43 (s, 3H), 2.39 - 2.27 (m, 1H), 2.22 - 2.13 (m, 1H), 1.63 (d, J = 7.2 Hz, 3H). (99.46% purity by HPLC)

[0494] Example 4

[0495] 4-(((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazoline-7-carbonitrile 4

[0496]

[0497] The first step: 4-Bromo-3-fluorobenzoic acid 4b

[0498] 3-Fluoro-4-bromotoluene 4a (5.0 g, 26.45 mmol) was dissolved in pyridine (20 mL) and water (20 mL). The mixture was heated to 90 °C, and potassium permanganate (16.7 g, 105.68 mmol) was added in portions over about 1 h. After the addition was complete, the reaction was continued for 2 h. TLC detection showed that the reaction was basically complete. The reaction solution was filtered through a pad of diatomaceous earth while hot. After the filtrate was cooled, it was adjusted to pH ≈ 3 with concentrated hydrochloric acid, extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a white solid, the title compound 4b (3.2 g, crude product), which was directly used in the next step.

[0499] The second step: 4-Bromo-5-fluoro-2-nitrobenzoic acid 4c

[0500] Compound 4b (3.2 g, crude) was dissolved in concentrated sulfuric acid (30 mL), and potassium nitrate (1.7 g, 16.82 mmol) was added portionwise. After addition, the reaction was carried out at room temperature for 2 hours, and TLC detection showed that the reaction was basically complete. Water was added to the reaction solution, and it was 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 4c (3.8 g, crude) as a pale yellow solid, which was directly used for the next step.

[0501] The third step: (S)-4-bromo-2-nitro-5-((tetrahydrofuran-3-yl)oxy)benzoic acid 4d

[0502] Compound 4c (3.8 g, crude) and compound IN-4c (3.8 g, 43.13 mmol) were dissolved in N,N-diisopropylethylamine (30 mL). Cesium carbonate (14.0 g, 42.97 mmol) was added at room temperature, and the mixture was heated to 100 °C and reacted for 4 hours. TLC detection showed that the reaction was basically complete. The reaction solution was cooled to room temperature, adjusted to about pH = 3 with dilute hydrochloric acid (3N), 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 4d (5.2 g, crude) as a yellow solid, which was directly used for the next step.

[0503] LC-MS: m / z = 330.0 [M-H] -

[0504] The fourth step: (S)-4-bromo-2-nitro-5-((tetrahydrofuran-3-yl)oxy)benzamide 4e

[0505] Compound 4d (5.2 g, crude) was dissolved in thionyl chloride (30 mL), and the mixture was heated to 80 °C and reacted for 3 hours. The reaction solution was cooled to room temperature, concentrated, and the residue was dissolved in tetrahydrofuran (50 mL). The solution was added dropwise to ammonia water (80 mL) cooled to about 0 °C. After addition, the reaction was carried out at 0 °C for 0.5 hour, and TLC detection showed that the reaction was complete. The reaction solution was diluted with water, 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 4e (3.6 g, overall yield of four steps 41%) as an off-white solid.

[0506] LC-MS: m / z = 331.1 [M+H] +

[0507] The fifth step: (S)-2-amino-4-bromo-5-((tetrahydrofuran-3-yl)oxy)benzamide 4f

[0508] Compound 4e (1.0 g, 3.02 mmol) was dissolved in ethanol (20 mL) and water (4 mL). Iron powder (676 mg, 12.10 mmol) and ammonium chloride (808 mg, 15.11 mmol) were added at room temperature, and the mixture was heated to 80 °C and reacted for 3 hours. The reaction was monitored by LCMS and found to be complete. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, the filtrate was concentrated, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 4f (830 mg, crude product), which was directly used in the next step.

[0509] LC-MS: m / z = 301.1 [M+H] +

[0510] Step 6 (S)-7-Bromo-2-methyl-6-((tetrahydrofuran-3-yl)oxy)quinazolin-4(3H)-one 4g

[0511] Compound 4f (830 mg, crude product) was dissolved in triethyl orthoformate (15 mL), and the mixture was heated to 150 °C and reacted for 5 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, and a solid precipitated. The solid was filtered, and the crude solid obtained was purified by silica gel column chromatography to obtain the off-white solid title compound 4g (490 mg, 50% yield over two steps).

[0512] LC-MS: m / z = 325.0 [M+H] +

[0513] Step 7 (S)-7-Bromo-4-chloro-2-methyl-6-((tetrahydrofuran-3-yl)oxy)quinazoline 4h

[0514] Compound 4g (490 mg, 1.51 mmol) was dispersed in 1,4-dioxane (5 mL). Pyridine (73 mg, 0.92 mmol) and phosphorus oxychloride (143 mg, 0.93 mmol) were added at room temperature, and the mixture was heated to 100 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was neutralized with sodium bicarbonate. The mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 4h (510 mg, crude product), which was directly used in the next step.

[0515] Step 8 7-Bromo-2-methyl-N-((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine 4i

[0516] Compound 4h (80 mg, crude) and intermediate IN-1 (41 mg, 0.18 mmol) were dissolved in dimethyl sulfoxide (2 mL). At room temperature, N,N-diisopropylethylamine (46 mg, 0.36 mmol) was added, and the mixture was heated to 100 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid titled compound 4i (70 mg, overall yield of two steps 74%).

[0517] LC-MS: m / z = 541.1 [M+H] +

[0518] Step 9 2-Methyl-4-(((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazoline-7-carbonitrile 4j

[0519] Compound 4i (70 mg, 0.13 mmol) was dissolved in N,N-diisopropylethylamine (3 mL). At room temperature, zinc cyanide (30 mg, 0.26 mmol) and tetrakis(triphenylphosphine)palladium (30 mg, 0.026 mmol) were added. The reaction system was purged with nitrogen three times, and then heated to 120 °C and reacted overnight. LCMS analysis showed that some starting material remained. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain a yellow solid mixture (starting material and product) titled compound 4j (50 mg, crude), which was directly used in the next step.

[0520] Step 10 4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazoline-7-carbonitrile 4

[0521] Compound 4j (50 mg, crude) was dissolved in ethanol (5 mL) and water (1 mL). At room temperature, ammonium chloride (30 mg, 0.56 mmol) and iron powder (30 mg, 0.54 mmol) were added successively, and the mixture was heated to 80 °C and reacted for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, filtered through celite, the filtrate was concentrated, the residue was added with water, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid titled compound 4 (8 mg, overall yield of two steps 14%).

[0522] LC-MS: m / z = 458.2 [M+H] +

[0523] 11H NMR (400 MHz, CD3OD) δ 7.91 (s, 1H), 7.86 (s, 1H), 7.01 - 6.95 (m, 2H), 6.81 (s, 1H), 5.65 (q, J = 7.2 Hz, 1H), 5.33 - 5.27 (m, 1H), 4.19 - 4.00 (m, 3H), 3.99 - 3.93 (m, 1H), 2.48 (s, 3H), 2.43 - 2.31 (m, 1H), 2.28 - 2.19 (m, 1H), 1.67 (d, J = 7.2 Hz, 3H). (96.53% purity by HPLC)

[0524] Example 5

[0525] N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((S)-2-methylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 5

[0526]

[0527] Step 1 (S)-4-(4-Carbamoyl-5-nitropyridin-2-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 5a

[0528] Compound 3c (2.0 g, 9.92 mmol) was dissolved in dimethyl sulfoxide (10 mL). At room temperature, (S)-4-N-Boc-2-methylpiperazine (2.2 g, 10.98 mmol) and N,N-diisopropylethylamine (2.6 g, 20.12 mmol) were added. The temperature was raised to 80 °C and the reaction was carried out for 5 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, water was added, and a solid precipitated. The solid was filtered, and the filter cake was washed and dried to obtain the yellow solid title compound 5a (4.2 g, crude), which was directly used in the next step.

[0529] LC-MS: m / z = 388.2 [M+Na] +

[0530] Step 2 (S)-4-(5-Amino-4-carbamoylpyridin-2-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 5b

[0531] Compound 5a (4.2 g, crude) was dissolved in ethanol (80 mL) and water (15 mL). Ammonium chloride (2.6 g, 48.61 mmol) and reduced iron powder (2.2 g, 39.39 mmol) were added at room temperature, and the mixture was heated to 80 °C and reacted for 3 hours. TLC detection showed that the reaction was basically complete. The reaction solution was filtered through diatomaceous earth while hot, the filtrate was concentrated, water was added to the residue, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 5b (3.5 g, crude), which was directly used for the next step.

[0532] LC-MS: m / z = 336.3 [M+H] +

[0533] The third step: (S)-tert-butyl 4-(4-hydroxy-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 5c

[0534] Compound 5b (3.5 g, crude) and p-toluenesulfonic acid (30 mg, 0.17 mmol) were dissolved in triethyl orthoacetate (30 mL), and the mixture was heated to 150 °C and reacted for 5 hours. TLC detection showed that the reaction was complete. The reaction solution was cooled to room temperature, dilute hydrochloric acid (20 mL, 1 N) was added, and the mixture was stirred at room temperature for 0.5 hour. The mixture was extracted with ethyl acetate, the organic phase was discarded, the aqueous phase was neutralized with sodium carbonate, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the yellow solid title compound 5c (1.7 g, overall yield of three steps 48%).

[0535] The fourth step: (S)-tert-butyl 4-(4-chloro-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 5d

[0536] Compound 5c (1.0 g, 2.78 mmol) was dissolved in 1,4-dioxane (10 mL). Triethylamine (845 mg, 8.35 mmol) was added at room temperature, and phosphorus oxychloride (1.28 g, 8.35 mmol) was added dropwise. After the addition was complete, the mixture was heated to 100 °C and reacted for 2 hours. TLC detection showed that the raw materials were basically completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the brown title compound 5d (1.0 g, crude), which was directly used for the next step.

[0537] LC-MS: m / z = 378.3 [M+H] +

[0538] Step 5 (S)-4-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 5e

[0539] Compound 5d (123 mg, crude) and intermediate IN-3 (50 mg, 0.26 mmol) were dissolved in dimethyl sulfoxide (2 mL). N,N-Diisopropylethylamine (67 mg, 0.52 mmol) was added at room temperature, and the mixture was heated to 90 °C and reacted for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 5e as a yellow solid (120 mg, yield 66% over two steps).

[0540] LC-MS: m / z = 531.3 [M+H] +

[0541] Step 6 N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((S)-2-methylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 5

[0542] Compound 5e (120 mg, 0.23 mmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (2 mL) was added, and the mixture was reacted at room temperature for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was concentrated, neutralized with saturated sodium bicarbonate solution, 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 as a yellow solid (90 mg, yield 91%).

[0543] LC-MS: m / z = 431.3 [M+H] +

[0544] 11H NMR (400 MHz, CD3OD) δ 8.67 (s, 1H), 7.59 (t, J = 7.2 Hz, 1H), 7.47 (t, J = 7.2 Hz, 1H), 7.30 (s, 1H), 7.22 (t, J = 7.6 Hz, 1H), 7.14 - 6.87 (m, 1H), 5.81 (q, J = 7.2 Hz, 1H), 4.71 - 4.57 (m, 1H), 4.00 (dd, J = 12.8, 2.4 Hz, 1H), 3.18 - 3.03 (m, 3H), 3.01 - 2.95 (m, 1H), 2.92 - 2.80 (m, 1H), 2.38 (s, 3H), 1.69 (d, J = 7.2 Hz, 3H), 1.24 (d, J = 6.8 Hz, 3H). (99.64% purity by HPLC)

[0545] Example 6

[0546] N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-6-((S)-2,4-dimethylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 6

[0547]

[0548] First step (S)-4-(4-Hydroxypyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 6a

[0549] Compound 5b (510 mg, 1.52 mmol) was dissolved in triethyl orthoformate (15 mL), p-toluenesulfonic acid (10 mg, 0.058 mmol) was added at room temperature, and the mixture was heated to 150 °C and reacted for 3 hours. TLC showed that the reaction was complete. The reaction solution was diluted with hydrochloric acid (1 N), extracted with ethyl acetate, the organic phase was discarded, the aqueous phase was neutralized with sodium carbonate, 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 6a (330 mg, yield 63%) as a yellow-brown solid.

[0550] Second step (S)-4-(4-Chloropyridido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 6b

[0551] Compound 6a (330 mg, 0.96 mmol) was dissolved in 1,4-dioxane (5 mL). Pyridine (440 mg, 5.56 mmol) was added at room temperature, and phosphorus oxychloride (377 mg, 2.46 mmol) was added dropwise. After the addition, the reaction mixture was heated to 100 °C and reacted for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, diluted by dropwise addition of dilute hydrochloric acid (10 mL, 1 N), extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the brown solid title compound 6b (350 mg, crude product), which was directly used for the next step.

[0552] The third step (S)-4-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)pyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 6c

[0553] Compound 6b (130 mg, crude product) and intermediate IN-3 (31 mg, 0.16 mmol) were dissolved in dimethyl sulfoxide (2 mL). N,N-Diisopropylethylamine (42 mg, 0.32 mmol) was added at room temperature, and the reaction mixture was heated to 100 °C and reacted for 2 hours. TLC showed that starting materials remained. Intermediate IN-3 (31 mg, 0.16 mmol) was added, and the reaction continued for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, water was added, and a solid precipitated. The solid was filtered, and the filter cake was purified by Prep-TLC to obtain the yellow solid title compound 6c (20 mg, yield 12%).

[0554] LC-MS: m / z = 517.3 [M+H] +

[0555] The fourth step N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-6-((S)-2-methylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 6d

[0556] Compound 6c (20 mg, 0.039 mmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (0.5 mL) was added at room temperature, and the reaction mixture was heated to 35 °C and reacted for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, adjusted to alkaline with saturated aqueous sodium bicarbonate, extracted with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 6d (20 mg, crude product), which was directly used for the next step.

[0557] The fifth step N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-6-((S)-2,4-dimethylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 6

[0558] Compound 6d (20 mg, crude) was dissolved in methanol (5 mL). Aqueous formaldehyde solution (20 mg, 0.25 mmol, 37%) and Pd / C (10 mg, 10%) were added at room temperature. The reaction mixture was heated to 30 °C under a hydrogen atmosphere and reacted for 3 hours. TLC showed that the reaction was complete. The reaction mixture was cooled to room temperature, filtered, the filter cake was washed, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid title compound 6 (10 mg, 59% yield in two steps).

[0559] LC-MS: m / z = 431.3 [M+H] +

[0560] 1 1H NMR (400 MHz, CD3OD) δ 8.80 (s, 1H), 8.28 (s, 1H), 7.59 (t, J = 7.6 Hz, 1H), 7.49 - 7.47 (m, 2H), 7.24 (t, J = 8.0 Hz, 1H), 7.14 - 6.89 (m, 1H), 5.81 (q, J = 6.8 Hz, 1H), 5.06 - 4.97 (m, 1H), 4.55 - 4.42 (m, 1H), 3.60 - 3.34 (m, 4H), 3.13 - 3.03 (m, 1H), 2.89 (s, 3H), 1.71 (d, J = 6.8 Hz, 3H), 1.36 (d, J = 7.2 Hz, 3H). (98.88% purity by HPLC)

[0561] Example 7

[0562] N 4 -((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-N 6 -((S)-tetrahydrofuran-3-yl)pyrido[3,4-d]pyrimidine-4,6-diamine 7

[0563]

[0564] The first step: Methyl 2-chloro-5-nitroisonicotinate 7a

[0565] Compound 3b (43.50 g, 0.21 mol) was dissolved in N,N-dimethylformamide (300 mL). Potassium carbonate (60.00 g, 0.43 mol) was added at room temperature, and methyl iodide (45.91 g, 0.32 mol) was added dropwise. After addition, the reaction mixture was stirred at room temperature for 2 hours. TLC showed that the reaction was complete. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the yellow solid title compound 7a (46.1 g, 99% yield).

[0566] LC-MS: m / z = 217.1 [M+H] +

[0567] Second step Methyl (S)-5-nitro-2-((tetrahydrofuran-3-yl)amino)isonicotinate 7b

[0568] Compound 7a (1.92 g, 8.87 mmol) was dissolved in tetrahydrofuran (30 mL). At room temperature, cesium carbonate (4.33 g, 13.29 mmol) and (S)-3-aminotetrahydrofuran 1k (850 mg, 9.76 mmol) were added. The temperature was raised to 30 °C and the reaction was carried out overnight. TLC showed that a small amount of the starting material remained unreacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 7b (2.28 g, crude product), which was directly used in the next step.

[0569] LC-MS: m / z = 268.1 [M+H] +

[0570] Third step Methyl (S)-5-amino-2-((tetrahydrofuran-3-yl)amino)isonicotinate 7c

[0571] Compound 7b (2.28 g, crude product) was dissolved in methanol (30 mL). Palladium / carbon (400 mg, 10%) was added. The temperature was raised to 30 °C under a hydrogen atmosphere and the reaction was carried out for 5 hours. TLC showed that the starting material had reacted completely. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with methanol. The filtrate was concentrated, and the crude product was purified by silica gel column chromatography to obtain the yellowish-brown solid title compound 7c (1.67 g, yield 80% over two steps).

[0572] LC-MS: m / z = 238.2 [M+H] +

[0573] Fourth step (S)-2-Methyl-6-((tetrahydrofuran-3-yl)amino)pyrido[3,4-d]pyrimidin-4-ol 7d

[0574] Compound 7c (700 mg, 2.95 mmol) was dissolved in acetonitrile (30 mL). The temperature was cooled to about 0 °C, and hydrogen chloride gas was introduced for 2 hours. After that, the temperature was raised to 80 °C and the reaction was carried out overnight. TLC showed that the starting material had reacted completely. The reaction solution was cooled to room temperature, adjusted to alkaline with sodium bicarbonate, filtered, and the filter cake was washed with water and then with methanol. The filter cake was added to toluene and concentrated to obtain the yellow solid title compound 7d (650 mg, crude product), which was directly used in the next step.

[0575] LC-MS: m / z = 247.1 [M+H] +

[0576] Step 5 (S)-4-Chloro-2-methyl-N-(tetrahydrofuran-3-yl)pyrido[3,4-d]pyrimidin-6-amine 7e

[0577] Compound 7d (500 mg, crude) was suspended in 1,4-dioxane (20 mL). At room temperature, N,N-diisopropylethylamine (1.05 g, 8.12 mmol) and phosphorus oxychloride (1.24 g, 8.09 mmol) were added. The mixture was heated to 90 °C and reacted for 4 h. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to 0 °C, quenched 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 7e (200 mg, crude) as a pale yellow solid, which was directly used for the next step.

[0578] LC-MS: m / z = 265.1 [M+H] +

[0579] Step 6 N 4 -((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-N 6 -((S)-tetrahydrofuran-3-yl)pyrido[3,4-d]pyrimidine-4,6-diamine 7

[0580] Compound 7e (78 mg, crude) was dissolved in dimethyl sulfoxide (3 mL). At room temperature, N,N-diisopropylethylamine (95 mg, 0.74 mmol) and intermediate IN-2 (60 mg, 0.29 mmol) were added. The mixture was heated to 80 °C and reacted for 8 h. TLC showed that the raw materials had reacted completely. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 7 (6 mg, overall yield of three steps 6%) as a pale yellow solid.

[0581] LC-MS: m / z = 433.3 [M+H] +

[0582] 11H NMR (400 MHz, CD3OD) δ 8.59 (s, 1H), 7.06 (s, 1H), 7.05 - 6.85 (m, 2H), 6.80 - 6.77 (m, 1H), 5.75 (q, J = 7.2 Hz, 1H), 4.45 - 4.37 (m, 1H), 4.10 - 3.98 (m, 2H), 3.94 - 3.87 (m, 1H), 3.73 (dd, J = 3.6, 8.8 Hz, 1H), 2.41 (s, 3H), 2.41 - 2.33 (m, 1H), 2.02 - 1.93 (m, 1H), 1.67 (d, J = 7.2 Hz, 3H). (97.91% purity by HPLC)

[0583] Example 8

[0584] N 4 -((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-N 6 -((S)-1-Methylpyrrolidin-3-yl)quinazoline-4,6-diamine 8

[0585]

[0586] The first step: 2-Amino-5-bromo-4-methoxybenzoic acid 8b

[0587] 2-Amino-4-methoxybenzoic acid 8a (15.0 g, 89.7 mmol) was dissolved in N,N-dimethylformamide (100 mL), cooled to 0 °C, and N-bromosuccinimide (17.6 g, 98.7 mmol) was added. The mixture was allowed to warm to room temperature and react for 2 hours. The reaction was monitored by TLC and found to be complete. Water was added to the reaction mixture, and the pH was adjusted to 3 with dilute hydrochloric acid (1 N). The mixture was extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium sulfite solution, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 8b (22.1 g, crude product) as a pale yellow solid, which was directly used in the next step.

[0588] The second step: Methyl 2-amino-5-bromo-4-methoxybenzoate 8c

[0589] Compound 8b (22.1 g, crude product) was dissolved in methanol (200 mL). Concentrated sulfuric acid (10 mL) was added dropwise at room temperature, and the temperature was then raised to 80 °C and stirred overnight. The reaction was monitored by TLC and found that the raw materials were basically completely reacted. The reaction mixture was cooled to room temperature, quenched with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 8c (19.5 g, overall yield of two steps 84%) as a pale yellow solid.

[0590] The third step: 6-Bromo-7-methoxy-2-methylquinazolin-4(3H)-one 8d

[0591] Compound 8c (19.5 g, 75.0 mmol) was dissolved in acetonitrile (150 mL). Methanesulfonic acid (40 mL) was added at room temperature, and the mixture was heated to 90 °C and stirred overnight. TLC showed that the raw materials had completely reacted. The reaction solution was concentrated, diluted with water, and the pH was adjusted to 8 - 9 with aqueous sodium hydroxide solution (2N). A solid precipitated out. It was filtered, and the filter cake was washed with water and dried to obtain the off-white solid title compound 8d (10.5 g, yield 52%).

[0592] Step 4: 6-Bromo-4-chloro-7-methoxy-2-methylquinazoline 8e

[0593] Compound 8d (1.00 g, 3.72 mmol) was dissolved in phosphorus oxychloride (6 mL). N,N-Dimethylformamide (0.1 mL) was added at room temperature, and the mixture was heated to 90 °C and reacted for 6 hours. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, poured into ice 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 pale yellow solid title compound 8e (535 mg, yield 50%).

[0594] LC-MS: m / z = 287.0 [M + H] +

[0595] Step 5: (R)-6-Bromo-7-methoxy-2-methyl-N-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)quinazolin-4-amine 8f

[0596] Compound 8e (500 mg, 1.74 mmol) and intermediate IN-1 (407 mg, 1.74 mmol) were dissolved in dimethyl sulfoxide (20 mL). N,N-Diisopropylethylamine (674 mg, 5.22 mmol) was added at room temperature, and the mixture was heated to 80 °C and stirred overnight. TLC showed that the reaction was complete. The reaction solution was cooled to room temperature, quenched with water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the yellow solid title compound 8f (810 mg, crude product), which was directly used in the next step.

[0597] LC-MS: m / z = 485.1 [M + H] +

[0598] Step 6 N 6 -((S)-1-Benzylpyrrolidin-3-yl)-7-methoxy-2-methyl-N 4 -((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)quinazoline-4,6-diamine 8h

[0599] Compound 8f (430 mg, 0.89 mmol) and (S)-1-benzyl-3-aminopyrrolidine 8g (312 mg, 1.77 mmol) were dissolved in toluene (20 mL). Tris(dibenzylideneacetone)dipalladium (162 mg, 0.18 mmol), 1,1′-binaphthalene-2,2′-bis(diphenylphosphine) (220 mg, 0.35 mmol) and cesium carbonate (577 mg, 1.77 mmol) were added at room temperature. The mixture was heated to 100 °C under nitrogen protection and stirred overnight. The reaction was monitored by TLC and the raw materials were completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the yellow solid title compound 8h (290 mg, yield 56.4%).

[0600] Step 7 N 4 -((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-N 6 -((S)-1-benzylpyrrolidin-3-yl)-7-methoxy-2-methylquinazoline-4,6-diamine 8i

[0601] Compound 8h (290 mg, 0.50 mmol) was dissolved in ethanol (20 mL) and water (4 mL). Iron powder (140 mg, 2.50 mmol) and ammonium chloride (134 mg, 2.50 mmol) were added at room temperature. The mixture was heated to 90 °C and stirred for 2 hours. The reaction was monitored by TLC and was completely reacted. The reaction solution was filtered through diatomaceous earth while hot, and the filtrate was concentrated. The residue was dissolved in ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow oily title compound 8i (275 mg, crude product), which was directly used for the next step.

[0602] Step 8 tert-Butyl (3-((R)-1-((6-(((S)-1-benzylpyrrolidin-3-yl)amino)-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carbamate 8j

[0603] Compound 8i (275 mg, crude product) was dissolved in tetrahydrofuran (32 mL). Sodium carbonate (159 mg, 1.50 mmol) and di-tert-butyl dicarbonate (218 mg, 1.00 mmol) were added, and the reaction was carried out at room temperature overnight. The reaction was monitored by TLC and was basically completely reacted. Water was added to the reaction solution, and the mixture was 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 Prep-TLC to obtain the pale yellow solid title compound 8j (70 mg, two-step yield 22%).

[0604] LC-MS: m / z = 651.3 [M+H] +

[0605] Step 9 tert-Butyl (3-((R)-1-((7-methoxy-2-methyl-6-(((S)-pyrrolidin-3-yl)amino)quinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carbamate 8k

[0606] Compound 8j (70 mg, 0.11 mmol) was dissolved in ethanol (10 mL), palladium on carbon (20 mg, 10%) was added, and the reaction was carried out at room temperature for 2 hours under a hydrogen atmosphere. The reaction was monitored by TLC and the starting material was completely reacted. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated to obtain the title compound 8k (50 mg, crude product) as a pale yellow oil, which was directly used in the next step.

[0607] Step 10 tert-Butyl (3-((R)-1-((7-methoxy-2-methyl-6-(((S)-1-methylpyrrolidin-3-yl)amino)quinazolin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)carbamate 8l

[0608] Compound 8k (50 mg, crude product) and aqueous formaldehyde solution (26 mg, 0.32 mmol, 37%) were dissolved in ethanol (10 mL), palladium on carbon (20 mg, 10%) was added, and the reaction was carried out at room temperature for 3 hours under a hydrogen atmosphere. The reaction was monitored by TLC and the starting material was completely reacted. The reaction mixture was filtered, and the filtrate was concentrated. The crude product was purified by Prep-TLC to obtain the title compound 8l (37 mg, overall yield of two steps: 60%) as a pale yellow solid.

[0609] Step 11 N 4 -((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-N 6 -((S)-1-methylpyrrolidin-3-yl)quinazolin-4,6-diamine 8

[0610] Compound 8l (37 mg, 0.06 mmol) was dissolved in methanol (1 mL), hydrochloric acid in methanol solution (3 mL, 4 M) was added, and the reaction was carried out at room temperature for 2 hours. The reaction was monitored by TLC and the starting material was completely reacted. The reaction mixture was neutralized with ammonia in methanol solution (4 M), the insoluble matter was filtered off, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 8 (20 mg, yield: 66%) as a pale yellow solid.

[0611] LC-MS: m / z = 475.3 [M+H] +

[0612] 11H NMR (400 MHz, CD3OD) δ 7.11 (s, 1H), 6.99 (s, 1H), 6.97 (s, 1H), 6.94 (s, 1H), 6.80 (s, 1H), 5.64 (q, J = 6.8 Hz, 1H), 4.32 - 4.25 (m, 1H), 3.98 (s, 3H), 3.00 - 2.86 (m, 2H), 2.70 - 2.66 (m, 1H), 2.63 - 2.55 (m, 1H), 2.55 - 2.47 (m, 1H), 2.45 (s, 6H), 1.82 - 1.74 (m, 1H), 1.64 (d, J = 6.8 Hz, 3H).

[0613] Example 9

[0614] 4 - (((R)-1-(3 - Amino - 5-(trifluoromethyl)phenyl)ethyl)amino)-6 - (((S)-tetrahydrofuran - 3 - yl)oxy)quinazoline - 7 - carbonitrile 9

[0615]

[0616] First step (S)-7 - Bromo - 6 - ((tetrahydrofuran - 3 - yl)oxy)quinazolin - 4(3H)-one 9a

[0617] Compound 4f (1.0 g, 3.32 mmol) and p - toluenesulfonic acid (20 mg, 0.12 mmol) were heated to 150 °C in triethyl orthoformate (10 mL) and reacted for 6 hours. The reaction was monitored by TLC and the starting materials were completely reacted. The reaction mixture was cooled to room temperature, and a solid precipitated. The solid was filtered, and the filter cake was washed and dried to obtain the off - white solid title compound 9a (540 mg, yield 52%).

[0618] LC - MS: m / z = 311.0 [M + H] +

[0619] Second step (S)-7 - Bromo - 4 - chloro - 6 - ((tetrahydrofuran - 3 - yl)oxy)quinazoline 9b

[0620] Compound 9a (300 mg, 0.96 mmol) was dispersed in 1,4 - dioxane (10 mL). Pyridine (228 mg, 2.88 mmol) and phosphorus oxychloride (442 mg, 2.88 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 2 hours. The reaction was monitored by TLC and the starting materials were completely reacted. The reaction mixture was cooled to room temperature, concentrated, and the residue was neutralized with saturated aqueous sodium bicarbonate. The mixture was extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep - TLC to obtain the yellow solid title compound 9b (73 mg, yield 23%).

[0621] Step 3 7-Bromo-N-((R)-1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine 9c

[0622] Compound 9b (73 mg, 0.22 mmol) and intermediate IN-1 (52 mg, 0.22 mmol) were dissolved in dimethyl sulfoxide (1 mL). At room temperature, N,N-diisopropylethylamine (57 mg, 0.44 mmol) was added, and the mixture was heated to 100 °C and reacted for 2 hours. TLC detection showed that the reaction was basically complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 9c as a yellow solid (85 mg, yield 73%).

[0623] LC-MS: m / z = 527.1 [M+H] +

[0624] Step 4 4-(((R)-1-(3-Nitro-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-7-carbonitrile 9d

[0625] Compound 9c (85 mg, 0.16 mmol) was dissolved in N,N-dimethylacetamide (3 mL). At room temperature, tetrakis(triphenylphosphine)palladium (18 mg, 0.016 mmol) and zinc cyanide (47 mg, 0.40 mmol) were added, and the mixture was heated to 140 °C and reacted for 7 hours under a nitrogen atmosphere. TLC detection showed that the raw materials had completely reacted. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 9d as a yellow solid (54 mg, yield 71%).

[0626] LC-MS: m / z = 474.2 [M+H] +

[0627] Step 5 4-(((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-7-carbonitrile 9

[0628] Compound 9d (54 mg, 0.11 mmol) was dissolved in ethanol (5 mL) and water (1 mL). Ammonium chloride (29 mg, 0.55 mmol) and reduced iron powder (25 mg, 0.44 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 2 hours. TLC detection showed that the raw materials had basically reacted completely. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, the filtrate was concentrated, water was added, and the mixture was 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 Prep-TLC to obtain the yellow solid title compound 9 (32 mg, yield 63%).

[0629] LC-MS: m / z = 444.2 [M+H] +

[0630] 1 1H NMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 8.02 (s, 1H), 7.90 (s, 1H), 6.96 (s, 2H), 6.81 (s, 1H), 5.60 (q, J = 6.8 Hz, 1H), 5.37 - 5.31 (m, 1H), 4.19 - 4.01 (m, 3H), 3.99 - 3.94 (m, 1H), 2.48 - 2.32 (m, 1H), 2.32 - 2.20 (m, 1H), 1.67 (d, J = 6.8 Hz, 3H).

[0631] Example 10

[0632] N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((R)-1-methylpiperidin-3-yl)pyrido[3,4-d]pyrimidin-4-amine 10-1

[0633] N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((S)-1-methylpiperidin-3-yl)pyrido[3,4-d]pyrimidin-4-amine 10-2

[0634]

[0635] The first step: 5-Amino-2-chloropyridine-4-carboxamide 10a

[0636] Compound 3c (1.0 g, 4.96 mmol) was dissolved in ethanol (20 mL) and water (4 mL). Ammonium chloride (1.3 g, 24.08 mmol) and reduced iron powder (1.1 g, 19.84 mmol) were added at room temperature, and the mixture was heated to 90 °C and reacted for 3 hours. TLC detection showed that the raw materials had completely reacted. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, water was added to the filtrate, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow-brown solid title compound 10a (680 mg, crude product), which was directly used for the next step.

[0637] The second step: tert-butyl 5-amino-4-carbamoyl-5′,6′-dihydro-[2,3′-bipyridine]-1′(2′H)-carboxylate 10c

[0638] Compound 10a (680 mg, crude product) was dissolved in 1,4-dioxane (8 mL) and water (2 mL). tert-Butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate 10b (1.5 g, 4.93 mmol), sodium carbonate (803 mg, 7.58 mmol) and [1,1′-bis(diphenylphosphino)ferrocene] palladium dichloride dichloromethane complex (50 mg, 0.061 mmol) were added successively at room temperature. Under nitrogen protection, the mixture was heated to 80 °C and reacted for 2 hours. TLC detection showed that the raw materials had basically reacted completely. The reaction solution was cooled to room temperature, water was added, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography to obtain the yellow solid title compound 10c (710 mg, overall yield of two steps 45%).

[0639] LC-MS: m / z = 319.2 [M+H] +

[0640] The third step: tert-butyl 3-(5-amino-4-carbamoylpyridin-2-yl)piperidine-1-carboxylate 10d

[0641] Compound 10c (710 mg, 2.23 mmol) was dissolved in ethanol (20 mL) and ethyl acetate (10 mL). Palladium on carbon (100 mg, 10%) was added, and the mixture was heated to 50 °C and reacted for 5 hours under a hydrogen atmosphere. LC-MS detection showed that the raw materials had completely reacted. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filtrate was concentrated to obtain the yellow solid title compound 10d (680 mg, crude product), which was directly used for the next step.

[0642] LC-MS: m / z = 321.3 [M+H] +

[0643] Step 4 tert-Butyl 3-(4-hydroxy-2-methylpyrido[3,4-d]pyrimidin-6-yl)piperidine-1-carboxylate 10e

[0644] Compound 10d (680 mg, crude) was dissolved in triethyl orthoacetate (6 mL), p-toluenesulfonic acid (10 mg) was added, and the mixture was heated to 150 °C for 6 h. The reaction was monitored by TLC and the starting material was completely consumed. The reaction mixture was cooled to room temperature, a small amount of dilute acid (1 N) was added and stirred for 0.5 h, then extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography to give the title compound 10e as a brown solid (188 mg, 24% yield over two steps).

[0645] Step 5 tert-Butyl 3-(4-chloro-2-methylpyrido[3,4-d]pyrimidin-6-yl)piperidine-1-carboxylate 10f

[0646] Compound 10e (160 mg, 0.44 mmol) was dissolved in 1,4-dioxane (5 mL), phosphorus oxychloride (409 mg, 2.6 mmol) and N,N-diisopropylethylamine (344 mg, 2.6 mmol) were added at room temperature, and the mixture was heated to 90 °C for 1.5 h. The reaction was monitored by TLC and the starting material was completely consumed. The reaction mixture was cooled to room temperature, water was added, then extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography to give the title compound 10f as a yellow solid (95 mg, 57% yield).

[0647] Step 6 (R)-tert-Butyl 3-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)piperidine-1-carboxylate 10g-1 & (S)-tert-Butyl 3-(4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)piperidine-1-carboxylate 10g-2

[0648] Compound 10f (75 mg, 0.20 mmol) and intermediate IN-3 (46 mg, 0.24 mmol) were dissolved in dimethyl sulfoxide (1.5 mL). N,N-Diisopropylethylamine (80 mg, 0.60 mmol) was added at room temperature, and the mixture was heated to 95 °C and reacted for 3 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by Prep-TLC to obtain the white solid title compound 10g (100 mg, yield 94%). Compound 10g was subjected to chiral resolution (Daicel AD-H, 30*250 mm, 5 μm, 30 mL / min, ethanol:Hexane = 5:95) to obtain the white solid compound 10g-1 (peak 1, RT 7.79 min) (30 mg, yield 28%) and the white solid compound 10g-2 (peak 2, RT 16.10 min) (48 mg, yield 45%). The configuration and properties of the compounds need to be further detected, and 10g-1 and 10g-2 are tentatively assigned the above configurations.

[0649] LC-MS: m / z = 516.3 [M+H] +

[0650] Step 7: N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((R)-piperidin-3-yl)pyrido[3,4-d]pyrimidin-4-amine 10h-1 & N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((S)-piperidin-3-yl)pyrido[3,4-d]pyrimidin-4-amine 10h-2

[0651] Compound 10g-1 (30 mg, 0.058 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC and was found to be complete. The reaction solution was concentrated, water was added, and the mixture was neutralized with saturated aqueous sodium carbonate. The mixture was extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the yellow solid title compound 10h-1 (25 mg, crude product) was obtained and directly used for the next step.

[0652] Compound 10g-2 (48 mg, 0.093 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC and was found to be complete. The reaction solution was concentrated, water was added, and the mixture was neutralized with saturated aqueous sodium carbonate. The mixture was extracted twice with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the yellow solid title compound 10h-2 (30 mg, crude product) was obtained and directly used for the next step.

[0653] The configuration and properties of the compound need to be further tested. Tentatively, 10h-1 and 10h-2 are the above configurations.

[0654] Step 8: N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((R)-1-methylpiperidin-3-yl)pyrido[3,4-d]pyrimidin-4-amine 10-1 & N-((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((S)-1-methylpiperidin-3-yl)pyrido[3,4-d]pyrimidin-4-amine 10-2 Compound 10h-1 (25 mg, crude) was dissolved in ethanol (2 mL), formaldehyde aqueous solution (4 drops, 37%) and palladium on carbon (10 mg, 10%) were added. Under a hydrogen atmosphere, the mixture was heated to 35 °C and reacted for 3 hours. TLC detection showed that the reaction was complete. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, the filtrate was concentrated, and the residue was purified by Prep-TLC to obtain the white solid title compound 10-1 (8 mg, two-step yield 32%).

[0655] Compound 10h-2 (30 mg, crude) was dissolved in ethanol (2 mL), formaldehyde aqueous solution (4 drops, 37%) and palladium on carbon (10 mg, 10%) were added. Under a hydrogen atmosphere, the mixture was heated to 35 °C and reacted for 3 hours. TLC detection showed that the reaction was complete. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, the filtrate was concentrated, and the residue was purified by Prep-TLC to obtain the white solid title compound 10-2 (21 mg, two-step yield 53%).

[0656] The configuration and properties of the compound need to be further tested. Tentatively, 10-1 and 10-2 are the above configurations.

[0657] 10-1:

[0658] LC-MS: m / z = 430.2 [M+H] +

[0659] 1 1H NMR (400 MHz, CD3OD) δ 8.96 (s, 1H), 8.26 (s, 1H), 7.66 (t, J = 7.2 Hz, 1H), 7.49 (t, J = 6.8 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.00 (t, J = 54.8 Hz, 1H), 5.86 (q, J = 7.2 Hz, 1H), 3.84 - 3.35 (m, 4H), 3.24 - 3.03 (m, 1H), 2.96 (s, 3H), 2.48 (s, 3H), 2.30 - 1.84 (m, 4H), 1.72 (d, J = 7.2 Hz, 3H).

[0660] 10-2:

[0661] LC-MS: m / z = 430.2 [M+H] +

[0662] 1 H NMR (400 MHz, CD3OD) δ 8.92 (s, 1H), 8.13 (s, 1H), 7.62 (t, J = 7.2 Hz, 1H), 7.49 (t, J = 6.8 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.00 (t, J = 54.8 Hz, 1H), 5.86 (q, J = 7.2 Hz, 1H), 3.28 - 3.18 (m, 2H), 3.12 - 3.04 (m, 1H), 2.65 - 2.55 (m, 1H), 2.50 (s, 3H), 2.46 (s, 3H), 2.40 - 2.28 (m, 1H), 2.16 - 2.06 (m, 1H), 1.98 - 1.74 (m, 3H), 1.72 (d, J = 7.2 Hz, 3H).

[0663] Example 11

[0664] (R)-4-((1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(4-methylpiperazin-1-yl)quinazoline-7-carbonitrile 11

[0665]

[0666] The first step: Methyl 4-bromo-5-fluoro-2-nitrobenzoate 11a

[0667] Compound 4c (2.6 g, 9.85 mmol) was dissolved in N,N-dimethylformamide (10 mL), methyl iodide (2.8 g, 19.73 mmol) and potassium carbonate (4.0 g, 28.94 mmol) were added, and the reaction was carried out at room temperature overnight. The reaction was monitored by TLC until completion. The reaction mixture was added with water (50 mL), extracted with ethyl acetate (100 mL), the organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 11a (3.0 g, crude product) as a pale yellow solid, which was directly used in the next step.

[0668] The second step: Methyl 4-bromo-5-(4-methylpiperazin-1-yl)-2-nitrobenzoate 11b

[0669] Compound 11a (3.0 g, crude) was dissolved in acetonitrile (20 mL). N-Methylmorpholine (1.6 g, 15.98 mmol) and potassium carbonate (2.0 g, 14.47 mmol) were added at room temperature, and the mixture was heated to 80 °C and reacted overnight. The reaction was monitored by TLC and found to be complete. The reaction mixture was diluted with water (50 mL), extracted with ethyl acetate (100 mL), the organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 11b (2.5 g, crude) as a pale yellow oil, which was directly used in the next step.

[0670] The third step: Methyl 2-amino-4-bromo-5-(4-methylpiperazin-1-yl)benzoate 11c

[0671] Compound 11b (2.5 g, crude) was dissolved in ethanol (20 mL). Iron powder (1.5 g, 26.86 mmol), ammonium chloride (1.5 g, 28.04 mmol) and water (2 mL) were added at room temperature, and the mixture was heated to 80 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was filtered through diatomaceous earth while hot, the filtrate was diluted with 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 11c (1.5 g, 47% yield over three steps) as a yellowish brown solid.

[0672] LC-MS: m / z = 328.1 [M+H] +

[0673] The fourth step: 7-Bromo-6-(4-methylpiperazin-1-yl)quinazolin-4-ol 11d

[0674] Compound 11c (1.5 g, 4.57 mmol) was dissolved in ethylene glycol monomethyl ether (10 mL). Formamidine acetate (3.0 g, 28.82 mmol) was added at room temperature, and the mixture was heated to 120 °C and reacted overnight. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, diluted with water, extracted with dichloromethane, 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 11d (1.3 g, 87% yield) as a pale yellow solid.

[0675] LC-MS: m / z = 323.1 [M+H] +

[0676] The fifth step: 7-Bromo-4-chloro-6-(4-methylpiperazin-1-yl)quinazoline 11e

[0677] Compound 11d (200 mg, 0.62 mmol) was dissolved in 1,4-dioxane (10 mL). At room temperature, N,N-diisopropylethylamine (2 mL) and phosphorus oxychloride (1 mL) were added. The mixture was heated to 90 °C and reacted overnight. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, concentrated, diluted with water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 11e as a yellow-brown solid (84 mg, crude product), which was directly used in the next step.

[0678] Step 6 (R)-7-bromo-6-(4-methylpiperazin-1-yl)-N-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)quinazolin-4-amine 11f

[0679] Compound 11e (84 mg, crude product) was dissolved in dimethyl sulfoxide (2 mL). At room temperature, N,N-diisopropylethylamine (70 mg, 0.54 mmol) and intermediate IN-1 (73 mg, 0.31 mmol) were added successively. The temperature was raised to 100 °C and the reaction was carried out for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 11f as a yellow-brown solid (130 mg, crude product), which was directly used in the next step.

[0680] Step 7 (R)-N-(1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)-7-bromo-6-(4-methylpiperazin-1-yl)quinazolin-4-amine 11g

[0681] Compound 11f (130 mg, crude product) was dissolved in ethanol (5 mL). At room temperature, iron powder (81 mg, 1.45 mmol), ammonium chloride (77 mg, 1.44 mmol) and water (1 mL) were added. The mixture was heated to 90 °C and reacted for 2 hours. The reaction was monitored by TLC and found that the raw materials were completely reacted. The reaction solution was filtered through diatomaceous earth while hot. The filtrate was diluted with water and 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 Prep-TLC to obtain the title compound as a pale yellow solid 11g (30 mg, overall yield of three steps: 10%).

[0682] LC-MS: m / z = 509.1 [M+H] +

[0683] Step 8 (R)-4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-6-(4-methylpiperazin-1-yl)quinazolin-7-carbonitrile 11

[0684] Compound 11g (30 mg, 0.059 mmol) was dissolved in N,N-dimethylacetamide (5 mL). Zinc cyanide (30 mg, 0.26 mmol) and tris(dibenzylideneacetone)dipalladium (10 mg, 0.011 mmol) were added at room temperature. Under nitrogen protection, the mixture was heated to 120 °C and reacted for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-HPLC to obtain the title compound 11 as a pale yellow solid (25 mg, yield 93%).

[0685] LC-MS: m / z = 456.2 [M+H] +

[0686] 1 1H NMR (400 MHz, CD3OD) δ 8.39 (s, 1H), 8.01 (s, 1H), 7.98 (s, 1H), 6.99 - 6.94 (m, 2H), 6.80 (s, 1H), 5.58 (q, J = 7.2 Hz, 1H), 3.38 - 3.34 (m, 4H), 2.84 - 2.75 (m, 4H), 2.45 (s, 3H), 1.67 (d, J = 7.2 Hz, 3H).

[0687] Example 12

[0688] 4-(((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazoline-7-carbonitrile 12

[0689]

[0690] First step: tert-Butyl (3-((R)-1-((7-bromo-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-yl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate 12a

[0691] Compound 4h (110 mg, crude) and intermediate IN-5 (60 mg, 0.20 mmol) were dissolved in dimethyl sulfoxide (2 mL). N,N-Diisopropylethylamine (52 mg, 0.40 mmol) was added at room temperature. The mixture was heated to 100 °C and reacted for 2 hours. The reaction was monitored by TLC and was found to be basically complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 12a as a yellow solid (63 mg, two-step yield 34%).

[0692] LC-MS: m / z = 611.2 [M+H]+

[0693] Step 2 tert-Butyl (3-((R)-1-((7-cyano-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-yl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate 12b

[0694] Compound 12a (63 mg, 0.10 mmol) was dissolved in N,N-dimethylacetamide (3 mL). Tetrakis(triphenylphosphine)palladium (18 mg, 0.016 mmol) and zinc cyanide (29 mg, 0.25 mmol) were added at room temperature. The reaction mixture was heated to 140 °C under nitrogen protection for 7 h, and the reaction was monitored by TLC until completion. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 12b (73 mg, crude), which was directly used in the next step.

[0695] Step 3 4-(((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-7-carbonitrile 12

[0696] Compound 12b (73 mg, crude) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (3 mL) was added, and the reaction was carried out at room temperature for 4 h. The reaction was monitored by TLC until completion. The reaction mixture was adjusted to basicity by dropwise addition of saturated aqueous sodium bicarbonate, and then extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-TLC to obtain the pale yellow solid title compound 12 (31 mg, 66% yield over two steps).

[0697] LC-MS: m / z = 458.2 [M+H] +

[0698] 1 1H NMR (400 MHz, CD3OD) δ 7.93 (s, 1H), 7.91 (s, 1H), 7.03 - 6.72 (m, 3H), 5.76 (q, J = 6.8 Hz, 1H), 5.37 - 5.31 (m, 1H), 4.19 - 4.01 (m, 3H), 3.99 - 3.94 (m, 1H), 2.45 (s, 3H), 2.43 - 2.35 (m, 1H), 2.32 - 2.22 (m, 1H), 1.67 (d, J = 6.8 Hz, 3H).

[0699] Example 13

[0700] N 4-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-7-methoxy-2-methyl-N 6 -((S)-tetrahydrofuran-3-yl)quinazoline-4,6-diamine 13

[0701]

[0702] Step 1 tert-Butyl ((R)-(3-(1-((6-bromo-7-methoxy-2-methylquinazolin-4-yl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate 13a

[0703] Compound 8e (600 mg, 2.09 mmol) and intermediate IN-5 (635 mg, 2.09 mmol) were dissolved in dimethyl sulfoxide (15 mL). At room temperature, N,N-diisopropylethylamine (810 mg, 6.27 mmol) was added. The temperature was raised to 80 °C and stirred overnight. The reaction was monitored by TLC and was found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 13a as a pale yellow solid (1.01 g, yield 87%).

[0704] LC-MS: m / z = 555.1 [M+H] +

[0705] Step 2 tert-Butyl (3-(difluoromethyl)-4-fluoro-5-((R)-1-((7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)amino)quinazolin-4-yl)amino)ethyl)phenyl)carbamate 13b

[0706] Compound 13a (100 mg, 0.18 mmol) and (S)-3-aminotetrahydrofuran 1k (32 mg, 0.36 mmol) were dissolved in toluene (15 mL). Tris(dibenzylideneacetone)dipalladium (33.0 mg, 0.036 mmol), 1,1′-binaphthalene-2,2′-bis(diphenylphosphine) (45 mg, 0.072 mmol), and cesium carbonate (117 mg, 0.36 mmol) were added. Under nitrogen protection, the temperature was raised to 100 °C and stirred for 10 hours. The reaction was monitored by TLC and the starting materials were found to have reacted almost completely. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with 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 as a yellow solid (70 mg, yield 69%).

[0707] LC-MS: m / z = 562.3 [M+H] +

[0708] Step 3 N 4-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-7-methoxy-2-methyl-N 6 -((S)-tetrahydrofuran-3-yl)quinazoline-4,6-diamine 13

[0709] Compound 13b (50.0 mg, 0.09 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 detection showed that the reaction was basically complete. The reaction solution was concentrated, the residue was neutralized with saturated aqueous sodium bicarbonate solution, extracted with dichloromethane, the organic phase was 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 as a pale yellow solid (20 mg, yield 49%).

[0710] LC-MS: m / z = 462.2 [M+H] +

[0711] 1 1H NMR (400 MHz, DMSO-d6) δ 7.20 (s, 1H), 7.03 - 6.85 (m, 3H), 6.80 - 6.75 (m, 1H), 5.78 (q, J = 6.8 Hz, 1H), 4.41 - 4.32 (m, 1H), 4.09 - 3.85 (m, 6H), 3.75 - 3.72 (m, 1H), 2.48 - 2.32 (m, 4H), 2.01 - 1.93 (m, 1H), 1.64 (d, J = 7.2 Hz, 3H).

[0712] Example 14

[0713] N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-6-((R)-3,4-dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidin-4-amine 14

[0714]

[0715] The first step (R)-1,2-dimethylpiperazine 14b

[0716] (R)-tert-Butyl 3,4-dimethylpiperazine-1-carboxylate 14a (1.5 g, 7.00 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (10 mL) was added, and the reaction was carried out at room temperature for 3 hours. TLC detection showed that the reaction was complete. The reaction solution was concentrated to obtain the title compound 14b as a yellow oil (1.5 g, crude product), which was directly used in the next step.

[0717] The second step (R)-benzyl 3,4-dimethylpiperazine-1-carboxylate 14c

[0718] Compound 14b (1.5 g, crude) was dispersed in tetrahydrofuran (20 mL) and water (10 mL). Sodium carbonate (6.9 g, 65.78 mmol) was added, and the mixture was stirred for 10 minutes. A solution of benzyl chloroformate (930 mg, 13.15 mmol) in tetrahydrofuran (5 mL) was added dropwise. The reaction was carried out at room temperature for 2 hours, and the reaction was monitored by TLC and found to be complete. The reaction mixture was filtered, and the filtrate was 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 14c (1.2 g, 70% yield in two steps) as a transparent oil.

[0719] LC-MS: m / z = 249.2 [M+H] +

[0720] The third step (R)-1,2-dimethylpiperazine 14d

[0721] Compound 14c (1.2 g, 4.80 mmol) was dissolved in tetrahydrofuran (15 mL). Palladium on carbon (120 mg, 10%) was added, and the reaction was carried out at 50 °C for 12 hours under a hydrogen atmosphere. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate. The filtrate was concentrated to obtain the title compound 14d (620 mg, crude) as a yellow oil, which was directly used in the next step.

[0722] The fourth step (R)-2-(3,4-dimethylpiperazin-1-yl)-5-nitroisonicotinic acid methyl ester 14e

[0723] Compound 14d (132 mg, crude) and methyl 2-chloro-5-nitroisonicotinate compound 7a (250 mg, 1.15 mmol) were dissolved in dimethyl sulfoxide (2 mL). N,N-Diisopropylethylamine (447 mg, 34.7 mmol) was added at room temperature, and the reaction was heated to 90 °C for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed 3 times with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 14e (270 mg, crude) as a brown solid, which was directly used in the next step.

[0724] LC-MS: m / z = 295.2 [M+H] +

[0725] The fifth step (R)-5-amino-2-(3,4-dimethylpiperazin-1-yl)isonicotinic acid methyl ester 14f

[0726] Compound 14e (270 mg, crude) was dispersed in ethanol / water (5 mL / 1.5 mL). Iron powder (258 mg, 4.6 mmol) and ammonium chloride (258 mg, 4.6 mmol) were added at room temperature, and the mixture was heated to 100 °C and reacted for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was filtered through diatomaceous earth while hot, and the filter cake was washed with ethanol. The filtrate was concentrated, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed three times 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 14f as a yellow solid (230 mg, 75% yield over three steps).

[0727] Step 6 (R)-6-(3,4-Dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidin-4-ol 14g

[0728] Compound 14f (130 mg, 0.49 mmol) was dissolved in acetonitrile (10 mL), cooled to 0 °C, and hydrogen chloride gas was bubbled through for 2 hours. The mixture was then heated to 80 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, concentrated, and neutralized with ammonia in methanol solution (15 mL, 4 M) at low temperature. The mixture was then concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 14g as a yellow solid (120 mg, 95% yield).

[0729] LC-MS: m / z = 274.2 [M+H] +

[0730] Step 7 (R)-4-Chloro-6-(3,4-dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidine 14h

[0731] Compound 14g (120 mg, 0.43 mmol) was dissolved in 1,4-dioxane (2 mL). N,N-Isopropylethylamine (113 mg, 0.87 mmol) was added at room temperature, and the mixture was cooled to 0 °C. Phosphorus oxychloride (134 mg, 0.87 mmol) was added dropwise. After the addition was complete, the mixture was heated to 90 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 14h as a yellow solid (126 mg, crude), which was directly used in the next step.

[0732] Step 8 N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-6-((R)-3,4-dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidin-4-amine 14

[0733] Compound 14h (126 mg, crude) and intermediate IN-2 (97 mg, 0.47 mmol) were dissolved in dimethyl sulfoxide (2 mL). N,N-Diisopropylethylamine (167 mg, 1.29 mmol) was added at room temperature, and the mixture was heated to 95 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 14 as a yellow solid (28 mg, 14% yield over two steps).

[0734] LC-MS: m / z = 460.3 [M+H] +

[0735] 1 1H NMR (400 MHz, CD3OD) δ 8.70 (s, 1H), 7.57 (s, 1H), 7.00 - 6.93 (m, 1H), 6.88 (t, J = 55.2 Hz, 1H), 6.75 - 6.73 (m, 1H), 5.76 (q, J = 7.2 Hz, 1H), 4.53 - 4.37 (m, 2H), 3.46 (d, J = 12.4 Hz, 1H), 3.38 - 3.32 (m, 1H), 3.19 - 3.00 (m, 3H), 2.81 (s, 3H), 2.45 (s, 3H), 1.67 (d, J = 6.8 Hz, 3H), 1.43 (d, J = 6.0 Hz, 3H).

[0736] Example 15

[0737] N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-6-((S)-2,4-dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidin-4-amine 15

[0738]

[0739] First step: tert-Butyl (S)-4-(4-(((R)-1-(5-amino-3-difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 15a

[0740] Compound 5d (91 mg, 0.24 mmol) was dissolved in dimethyl sulfoxide (2 mL). Intermediate IN-2 (50 mg, 0.24 mmol) and N,N-diisopropylethylamine (62 mg, 0.48 mmol) were added at room temperature. The temperature was raised to 100 °C and the reaction was carried out for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid title compound 15a (50 mg, yield 38%).

[0741] Second step: tert-butyl (S)-4-(4-(((R)-1-(5-acetamido-3-difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 15b

[0742] Compound 15a (50 mg, 0.092 mmol) was dissolved in dichloromethane (5 mL). Pyridine (22 mg, 0.28 mmol) and acetic anhydride (14 mg, 0.14 mmol) were added at room temperature. The temperature was raised to 30 °C and the reaction was carried out for 4 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, concentrated, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the yellow solid title compound 15b (85 mg, crude product), which was directly used in the next step.

[0743] LC-MS: m / z = 588.3 [M+H] +

[0744] Third step: N-(3-(difluoromethyl)-4-fluoro-5-((R)-1-(2-methyl-6-((S)-2-methylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)phenyl)acetamide 15c

[0745] Compound 15b (85 mg, crude product) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (1 mL) was added at room temperature. The temperature was raised to 30 °C and the reaction was carried out for 2 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, concentrated, and the residue was neutralized with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the yellow solid title compound 15c (55 mg, crude product), which was directly used in the next step.

[0746] LC-MS: m / z = 488.3 [M+H] +

[0747] Step 4 N-(3-(Difluoromethyl)-5-((R)-1-((6-((S)-2,4-Dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)-4-fluorophenyl)acetamide 15d

[0748] Compound 15c (55 mg, crude) was dissolved in ethanol (5 mL), formaldehyde aqueous solution (30 mg, 0.37 mmol, 37%) and palladium on carbon (10 mg, 10%) were added, and the reaction was carried out at room temperature for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated to obtain a yellow solid of the title compound 15d (66 mg, crude), which was directly used for the next step.

[0749] Step 5 N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-6-((S)-2,4-dimethylpiperazin-1-yl)-2-methylpyrido[3,4-d]pyrimidin-4-amine 15

[0750] Compound 15d (66 mg, crude) was dispersed in ethanol (2 mL) and aqueous sodium hydroxide solution (2.0 mL, 8.00 mmol, 4 M), and the mixture was heated to 90 °C and reacted for 4 hours. The reaction was monitored by TLC and the raw materials were found to have reacted completely. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain a yellow solid of the title compound 15 (12 mg, 29% yield over four steps).

[0751] LC-MS: m / z = 460.3 [M+H] +

[0752] 1 H NMR (400 MHz, CD3OD) δ 8.60 (s, 1H), 7.33 (s, 1H), 7.00 - 6.55 (m, 3H), 5.65 (q, J = 6.8 Hz, 1H), 4.77 - 4.67 (m, 2H), 4.14 (d, J = 14.2 Hz, 1H), 3.18 - 3.02 (m, 2H), 2.75 - 2.64 (m, 1H), 2.58 - 2.47 (m, 4H), 2.34 (s, 3H), 1.56 (d, J = 7.2 Hz, 3H), 1.20 (d, J = 6.8 Hz, 3H).

[0753] Example 16

[0754] N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)pyrido[3,4-d]pyrimidin-4-amine 16

[0755]

[0756] Step 1: (1S,4S)-tert-butyl 5-(4-carbamoyl-5-nitropyridin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate 16b

[0757] Compound 3c (500 mg, 2.50 mmol) and (1S,4S)-tert-butyl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate 16a (500 mg, 2.50 mmol) were dissolved in dimethyl sulfoxide. N,N-Diisopropylethylamine (967 mg, 7.50 mmol) was added at room temperature, and the mixture was heated to 95 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with 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 as a brown solid (1.0 g, yield 94%).

[0758] Step 2: (1S,4S)-tert-butyl 5-(5-amino-4-carbamoylpyridin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate 16c

[0759] Compound 16b (1.0 g, 2.7 mmol) was dissolved in ethanol (20 mL) and water (5 mL). Reductive iron powder (770 mg, 13.7 mmol) and ammonium chloride (770 mg, 13.7 mmol) were added at room temperature, and the mixture was heated to 100 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was filtered through diatomaceous earth while hot, the filter cake was washed, the filtrate was concentrated, water was added, and the mixture was extracted with ethyl acetate three times. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the title compound 16c as a brown solid (950 mg, crude product), which was directly used in the next step.

[0760] Step 3: (1S,4S)-tert-butyl 5-(2-methyl-4-oxo-3,4-dihydropyrido[3,4-d]pyrimidin-6-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate 16d

[0761] The crude compound 16c (950 mg) was dissolved in triethyl orthoacetate (9 mL). p-Toluenesulfonic acid (30 mg, 0.17 mmol) was added at room temperature, and the mixture was heated to 150 °C and reacted for 2 hours. The reaction was monitored by TLC and found that the raw materials were completely reacted. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate three times. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 16d as a brown solid (570 mg, overall yield of two steps 58%).

[0762] LC-MS: m / z = 358.2 [M+H] +

[0763] Step 4: tert-Butyl (1S,4S)-5-(4-chloro-2-methylpyrido[3,4-d]pyrimidin-6-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate 16e

[0764] Compound 16d (200 mg, 0.56 mmol) was dissolved in 1,4-dioxane (3 mL). Phosphorus oxychloride (257 mg, 1.6 mmol) and N,N-diisopropylethylamine (217 mg, 1.6 mmol) were added at room temperature. The mixture was heated to 90 °C and reacted for 1.5 h. The reaction was monitored by TLC and the starting material was completely consumed. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to give the title compound 16e as a yellow solid (122 mg, yield 58%).

[0765] Step 5: tert-Butyl (1S,4S)-5-(4-(((R)-1-(3-(difluoromethyl)-2-fluoro-5-((isopropoxycarbonyl)amino)phenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate 16f

[0766] Compound 16e (122 mg, 0.32 mmol) was dissolved in dimethyl sulfoxide (2 mL). Intermediate IN-6 (93 mg, 0.32 mmol) and N,N-diisopropylethylamine (42 mg, 0.96 mmol) were added at room temperature. The mixture was heated to 95 °C and reacted for 1.5 h. The reaction was monitored by TLC and was complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to give the title compound 16f as a yellow solid (150 mg, crude product), which was directly used in the next step.

[0767] Step 6: Isopropyl (3-((R)-1-((6-((1S,4S)-2,5-diazabicyclo[2.2.1]heptan-2-yl)-2-methylpyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate 16g

[0768] Compound 16f (150 mg, crude) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2.5 mL) was added, and the reaction was carried out at room temperature for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was concentrated, quenched with ice water, adjusted to alkaline with saturated aqueous sodium bicarbonate, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid titled compound 16g (137 mg, crude), which was directly used for the next step.

[0769] Step 7: Isopropyl (3-(difluoromethyl)-4-fluoro-5-((R)-1-((2-methyl-6-((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)pyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)phenyl)carbamate 16h

[0770] Compound 16g (137 mg, crude) was dissolved in ethanol (3 mL), formaldehyde solution (167 mg, 0.20 mmol, 37%) and palladium on carbon (10 mg, 10%) were added, and the reaction was heated to 30 °C in a hydrogen atmosphere for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, filtered through diatomaceous earth, the filter cake was washed, and the filtrate was concentrated to obtain the yellow solid titled compound 16h (137 mg, crude), which was directly used for the next step.

[0771] LC-MS: m / z = 544.3 [M+H] +

[0772] Step 8: N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)pyrido[3,4-d]pyrimidin-4-amine 16

[0773] Compound 16h (137 mg, crude) was dissolved in ethanol (2 mL), aqueous sodium hydroxide solution (2 mL, 6 mmol, 3 M) was added at room temperature, and the reaction was heated to 95 °C for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate three times. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid titled compound 16 (45 mg, overall yield of four steps 30%).

[0774] LC-MS: m / z = 458.2 [M+H] +

[0775] 11H NMR (400 MHz, CD3OD) δ 8.55 (s, 1H), 7.07 (s, 1H), 6.95 - 6.66 (m, 3H), 5.67 (q, J = 7.2 Hz, 1H), 4.77 (s, 1H), 3.77 (s, 1H), 3.60 (d, J = 10.4 Hz, 1H), 3.51 (dd, J = 10.4, 2.0 Hz, 1H), 2.98 - 2.88 (m, 2H), 2.50 (s, 3H), 2.34 (s, 3H), 2.08 (d, J = 10.4 Hz, 1H), 1.97 (d, J = 10.0 Hz, 1H), 1.59 (d, J = 7.2 Hz, 3H).

[0776] Example 17

[0777] N-((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-6-((S)-2,4-dimethylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 17

[0778]

[0779] Step 1 (S)-tert-Butyl 4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 17a

[0780] Compound 6b (63 mg, 0.17 mmol) and Intermediate IN-1 (41 mg, 0.17 mmol) were dissolved in dimethyl sulfoxide (1 mL). N,N-Diisopropylethylamine (67 mg, 0.52 mmol) was added at room temperature, and the mixture was heated to 100 °C and reacted for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 17a (130 mg, crude), which was directly used in the next step.

[0781] Step 2 (S)-tert-Butyl 4-(4-(((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 17b

[0782] Compound 17a (130 mg, crude) was dispersed in ethanol / water (10 mL / 2 mL). Iron powder (64 mg, 1.15 mmol) and ammonium chloride (64 mg, 1.15 mmol) were added at room temperature, and the mixture was heated to 100 °C and reacted for 3 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was filtered through diatomaceous earth while hot, and the filter cake was washed. The filtrate was concentrated, water was added, and the mixture was extracted 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 yellow solid title compound 17b (87 mg, 94% yield over two steps).

[0783] LC-MS: m / z = 532.3 [M+H] +

[0784] The third step: (S)-tert-butyl 4-(4-(((R)-1-(3-acetamido-5-(trifluoromethyl)phenyl)ethyl)amino)pyrido[3,4-d]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylate 17c

[0785] Compound 17b (87 mg, 0.16 mmol) was dissolved in dichloromethane (3 mL). N,N-Diisopropylethylamine (42 mg, 0.32 mmol) and acetic anhydride (20 mg, 0.19 mmol) were added at room temperature, and the mixture was heated to 30 °C and reacted for 4 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, concentrated, water was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the yellow solid title compound 17c (100 mg, crude), which was directly used in the next step of the reaction.

[0786] The fourth step: N-(3-((R)-1-((6-((S)-2-methylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 17d

[0787] Compound 17c (100 mg, crude) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (0.5 mL) was added at room temperature, and the mixture was heated to 30 °C and reacted for 3 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was concentrated, water was added, and the mixture was extracted 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 yellow solid title compound 17d (77 mg, 99% yield over two steps).

[0788] The fifth step: N-(3-((R)-1-((6-((S)-2,4-dimethylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)-5-(trifluoromethyl)phenyl)acetamide 17e

[0789] Compound 17d (77 mg, 0.16 mmol) was dissolved in ethanol (3 mL). Aqueous formaldehyde solution (55 mg, 0.67 mmol, 37%) and palladium on carbon (10 mg, 10%) were added at room temperature. The mixture was heated to 35 °C and reacted for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction mixture was cooled to room temperature, filtered through celite, and the filtrate was concentrated to obtain the title compound 17e (78 mg, crude product) as a yellow oil, which was directly used in the next step.

[0790] LC-MS: m / z = 488.3 [M+H] +

[0791] Step 6: N-((R)-1-(3-Amino-5-(trifluoromethyl)phenyl)ethyl)-6-((S)-2,4-dimethylpiperazin-1-yl)pyrido[3,4-d]pyrimidin-4-amine 17

[0792] Compound 17e (78.0 mg, crude product) was dissolved in ethanol (2 mL). Aqueous sodium hydroxide solution (2.0 mL, 8.0 mmol, 4 M) was added at room temperature. The mixture was heated to 75 °C and reacted overnight. The reaction was monitored by TLC and found to be complete. The reaction mixture was concentrated, extracted with ethyl acetate, washed three times with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the title compound 17 (10 mg, overall yield of two steps 13%) as a yellow solid.

[0793] LC-MS: m / z = 446.2 [M+H] +

[0794] 1 1H NMR (400 MHz, CD3OD) δ 8.74 (s, 1H), 8.24 (s, 1H), 7.30 (s, 1H), 6.95 (s, 2H), 6.81 (s, 1H), 5.54 (q, J = 6.8 Hz, 1H), 4.80 - 7.83 (m, 1H), 4.12 (d, J = 12.8 Hz, 1H), 3.29 - 3.20 (m, 1H), 3.03 (d, J = 11.2 Hz, 1H), 2.93 (d, J = 11.2 Hz, 1H), 2.44 (d, J = 8.0 Hz, 1H), 2.39 (s, 3H), 2.30 - 2.20 (m, 1H), 1.65 (d, J = 6.8 Hz, 3H), 1.26 (d, J = 6.8 Hz, 3H).

[0795] Example 18

[0796] N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-(6-methyl-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyrido[3,4-d]pyrimidin-4-amine 18

[0797]

[0798] Step 1: tert-Butyl 3-(4-carbamoyl-5-nitropyridin-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate 18b

[0799] Compound 3c (1.0 g, 4.96 mmol) was dissolved in dimethyl sulfoxide (3 mL). At room temperature, tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate 18a (990 mg, 4.99 mmol) and N,N-diisopropylethylamine (1.9 g, 14.70 mmol) were added. The mixture was heated to 90 °C and reacted for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 18b as a yellow solid (1.7 g, yield 94%).

[0800] LC-MS: m / z = 364.2 [M+H] +

[0801] Step 2: tert-Butyl 3-(5-amino-4-carbamoylpyridin-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate 18c

[0802] Compound 18b (1.7 g, 4.68 mmol) was dispersed in ethanol (20 mL) and water (5 mL). At room temperature, iron powder (1.5 g, 26.86 mmol) and ammonium chloride (1.4 g, 26.17 mmol) were added. The mixture was heated to 90 °C and reacted for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was filtered while hot, the filter cake was washed with ethanol, the filtrate was concentrated, water was added, and the mixture was extracted 3 times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 18c as a yellow solid (1.4 g, yield 90%).

[0803] LC-MS: m / z = 334.2 [M+H] +

[0804] Step 3: tert-Butyl 3-(2-methyl-4-oxo-3,4-dihydropyrido[3,4-d]pyrimidin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate 18d

[0805] Compound 18c (1.4 g, 4.20 mmol) was dispersed in triethyl orthoacetate (16 mL). p-Toluenesulfonic acid (60 mg, 0.35 mmol) was added at room temperature, and the mixture was heated to 150 °C and reacted for 3 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, diluted hydrochloric acid (10 mL, 1 N) was added and stirred for 10 minutes. The mixture was extracted with ethyl acetate, and the organic phase was discarded. The aqueous phase was neutralized with saturated aqueous sodium carbonate, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound 18d as a yellow solid (805 mg, yield 54%).

[0806] Step 4: tert-Butyl 3-(4-chloro-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate 18e

[0807] Compound 18d (100 mg, 0.29 mmol) was dissolved in 1,4-dioxane (3 mL). N,N-Diisopropylethylamine (108 mg, 0.84 mmol) was added, and the temperature was cooled to 0 °C. Phosphorus oxychloride (128 mg, 0.83 mmol) was added dropwise. After addition, the mixture was heated to 90 °C and reacted for 3 hours. The reaction was monitored by TLC and the starting material was found to be completely reacted. The reaction mixture was cooled to room temperature, ice water was added, and the mixture was extracted 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 18e as a yellow solid (54 mg, yield 50%).

[0808] Step 5: tert-Butyl 3-(4-(((R)-1-(3-(difluoromethyl)-2-fluoro-5-((isopropoxycarbonyl)amino)phenyl)ethyl)amino)-2-methylpyrido[3,4-d]pyrimidin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate 18f

[0809] Compound 18e (54 mg, 0.14 mmol) was dissolved in dimethyl sulfoxide (2 mL). Intermediate IN-6 (42 mg, 0.14 mmol) and N,N-diisopropylethylamine (67 mg, 0.52 mmol) were added at room temperature, and the mixture was heated to 95 °C and reacted for 1.5 hours. The reaction was monitored by TLC and was found to be complete. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the title compound 18f as a green solid (100 mg, crude product), which was directly used in the next step.

[0810] LC-MS: m / z = 630.3 [M+H] +

[0811] Step 6 Isopropyl (3-((1R)-1-((6-(3,6-diazabicyclo[3.1.1]heptan-3-yl)-2-methylpyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)-5-(difluoromethyl)-4-fluorophenyl)carbamate 18 g

[0812] Compound 18f (100 mg, crude) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at room temperature for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was concentrated, water was added, the pH was adjusted to neutral with saturated aqueous sodium bicarbonate solution, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain the yellow solid title compound 18g (90 mg, crude), which was directly used in the next step.

[0813] LC-MS: m / z = 530.3 [M+H] +

[0814] Step 7 Isopropyl (3-(difluoromethyl)-4-fluoro-5-((1R)-1-((2-methyl-6-(6-methyl-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyrido[3,4-d]pyrimidin-4-yl)amino)ethyl)phenyl)carbamate 18h

[0815] Compound 18g (90 mg, crude) was dissolved in ethanol (3 mL), formaldehyde solution (55 mg, 0.68 mmol, 37%) and palladium on carbon (10 mg, 10%) were added, and the reaction was heated to 30 °C in a hydrogen atmosphere for 2 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, the filtrate was concentrated, and the crude product was purified by Prep-TLC to obtain the white solid title compound 18h (30 mg, 19% yield over four steps).

[0816] LC-MS: m / z = 544.3 [M+H] +

[0817] Step 8 N-((R)-1-(5-Amino-3-(difluoromethyl)-2-fluorophenyl)ethyl)-2-methyl-6-(6-methyl-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyrido[3,4-d]pyrimidin-4-amine 18

[0818] Compound 18h (30 mg, 0.055 mmol) was dissolved in ethanol (2 ml), aqueous sodium hydroxide solution (2 mL, 6.00 mmol, 3N) was added at room temperature, and the reaction was heated to 95 °C for 3 hours. The reaction was monitored by TLC and found to be complete. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted 3 times with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC to obtain the yellow solid title compound 18 (10 mg, 40% yield).

[0819] LC-MS: m / z = 458.3 [M+H] +

[0820] 1 H NMR (400 MHz, CD3OD) δ 8.73 (s, 1H), 7.33 (s, 1H), 7.03 - 6.92 (m, 1H), 6.88 (t, J = 55.2 Hz, 1H), 6.80 - 6.72 (m, 1H), 5.77 (q, J = 6.8 Hz, 1H), 4.51 - 3.85 (m, 6H), 3.25 - 3.10 (m, 1H), 2.97 - 2.78 (m, 1H), 2.56 - 2.46 (m, 2H), 2.44 (s, 3H), 2.11 - 1.93 (s, 1H), 1.68 (d, J = 7.2 Hz, 3H).

[0821] Test Example 1 Inhibitory Effect of the Compound on the Proliferation of K-562 Cells IC 50 Determination

[0822] The human chronic myelogenous leukemia cell line K-562 (CCL-243) used in the present invention was purchased from American Type Culture Collection (ATCC). The cells were grown in RPMI 1640 medium containing 10% fetal bovine serum (FBS) and 1% double antibody at 37°C in a 5% CO2 environment.

[0823] The inhibitory effect of the compound on the proliferation of K-562 cells cultured in vitro was determined by the following method:

[0824] 1) Cell seeding: K-562 cells in the logarithmic growth phase with good status were seeded at 20,000 cells / well and 90 μL into a 96-well plate and cultured at 37°C and 5% CO2 for 24 hours.

[0825] 2) Drug addition: The compound to be tested was serially diluted with complete medium. 10 μL of the diluted compound was added to 90 μL of the cells to make the final concentration of the compound 10,000, 3,000, 1,000, 300, 100, 30, 10, 3, 1 nM. At the same time, a corresponding solvent control was set. It was placed in a cell culture incubator at 37°C and 5% CO2 and cultured for 96 hours.

[0826] 3) Detection: 10 μL of 5 mg / mL MTT working solution (ABCONE, M9609) was added to each well. After acting at 37°C for 4 hours, triple solution (10% SDS, 0.5% isopropanol, 0.1 mol / L HCL) was added until the cell lysate was completely dissolved. The OD570 and OD690 values were read using a TECAN SPARK microplate reader.

[0827] 4) Calculation: Calculate the cell growth inhibition rate according to the following formula:

[0828] Inhibition rate = (control well OD570nm-OD690nm - treated well OD570nm-OD690nm ) / control well OD570nm-OD690nm × 100%

[0829] Use Graphpad prism 5.0 software to calculate the IC 50 value according to the compound concentration and the corresponding inhibition rate. The test results are shown in Table 1.

[0830] Compound Number K-562 Compound Number K-562 BI-3406 35.2 10-1 303.8 1 36.4 10-2 147.8 2 111.0 11 40.7 3 138.9 12 210.25 4 203.4 13 22.5 5 89.5 14 35.6 6 129.1 15 19.0 7 92.4 16 143.7 8 153.4 17 32.7 9 105.7 18 10.0

[0831] Conclusion: The compounds in the embodiments of the present invention have a proliferation inhibitory effect on K-562 cells, and the activities of multiple compounds are comparable to that of BI-3402.

[0832] Test Example 2 Effect of the compound on the phosphorylation level of the KRAS downstream signaling molecule ERK1 / 2 in K-562 cells

[0833] The effect of the compound of the present invention on the phosphorylation level of ERK1 / 2 in K-562 cells was detected by the following method:

[0834] 1) Cell seeding: Take K-562 cells in good condition in the logarithmic growth phase and seed them into a six-well plate at 1*10 6 cells / well, and culture overnight at 37°C and 5% CO2.

[0835] 2) Drug addition: After gradient diluting the compound to be tested with complete medium, add it to the cells so that the final concentration of the compound is 1000, 100, 10, 1 nM. Incubate in a 37°C, 5% CO2 cell culture incubator for 24 hours.

[0836] 3) Protein sample preparation: Collect the cell suspension, centrifuge at 500g for 5 minutes, discard the supernatant, wash 3 times with PBS, and lyse the cells with 100 μL of 1× SDS gel loading buffer (50 mM Tris-HCl (pH 6.8), 100 mM DTT, 2% SDS, 10% glycerol, 0.1% bromophenol blue). Heat the cell lysate at 100°C for 10 minutes for denaturation.

[0837] 4) Western blot: Perform SDS-PAGE electrophoresis on protein samples. After electrophoresis, transfer the proteins to a PVDF membrane using a wet transfer system. Place the PVDF membrane in a blocking solution (5% skim milk diluted in TBS / T) and block at room temperature for 1 hour. Then, perform the primary and secondary antibody reactions. After washing the membrane, develop the color using the Immobilon Western HRP Substrate luminal reagent, and take a picture with a 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).

[0838] The results of the effect of the compound on the phosphorylation level of ERK1 / 2 in K-562 cells are shown in Figure 1 .

[0839] Conclusion: The compound in the embodiment of the present invention has a significant inhibitory effect on the phosphorylation of ERK1 / 2 in K-562 cells, and the inhibitory activity shows a concentration gradient dependence.

[0840] Test Example 3 Inhibitory Activity of the Compound on SOS1

[0841] Experimental procedure:

[0842] Treatment of the compound

[0843] Prepare a compound with a final concentration 400 times higher. For example, if the final concentration to be detected is 5 μM, prepare a 400-fold concentration, that is, 2 mM. Use an automatic micropipette to serially dilute the compound into the set number of concentration points.

[0844] Transfer the compound to a 384-well reaction plate

[0845] Use an ultrasonic nanoliter liquid handling system to transfer 50 nL of the diluted compound from an Echo 384-well plate to a 384-well reaction plate. Transfer 50 nL of 100% DMSO for both the negative control and the positive control.

[0846] Prepare and transfer a 4-fold Tag1-SOS1 solution

[0847] Prepare a 4-fold Tag1-SOS1 solution using the Diluent provided in the kit, and transfer 5 μL to a 384-well reaction plate. For the negative control wells, transfer 5 μL of Diluent to replace the enzyme solution, and centrifuge at 1000 rpm for 1 minute.

[0848] Prepare a 4-fold Tag2-KRAS G12C solution

[0849] Prepare a 4-fold Tag2-KRAS G12C solution using the Diluent provided in the kit, transfer 5 μl to a 384-well reaction plate, and centrifuge at 1000 rpm for 1 minute.

[0850] Transfer 2-fold detection solution

[0851] Prepare 2-fold Anti-Tag1-Tb3+ and Anti-Tag2-XL665 solutions using the Detection Buffer provided in the kit, transfer 10 μl to a 384-well reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 60 minutes.

[0852] Reading

[0853] Read the fluorescence signal value (Ex665 / Em615) using an Envision microplate reader.

[0854] Inhibition rate calculation and IC50 fitting

[0855] Copy the values from the plate reader, where the maximum value refers to the reading of the positive control and the minimum value refers to the reading of the negative control. Inhibition rate (%) = (maximum value - sample value) / (maximum value - minimum value) × 100%.

[0856] Import the data into MS Excel and fit the IC50 value using the XLFit excel add-in version 5.4.0.8;

[0857] Compound Number IC50 (nM) Compound Number IC50 (nM) Compound Number IC50 (nM) 1 5.7 2 6.7 3 10.1 5 6.6 6 7.5 8 5.4 9 8.5 10-2 13.0 11 11 12 6.5 13 4.0 14 5.6 15 4.7 16 5.4 17 5.1 18 4.2

[0858] Fitting formula: Y = Bottom + (Top - Bottom) / (1 + (IC50 / X)^HillSlope)

[0859] Test Example 4 Stability Experiment of Compounds in Mouse and Human Liver Microsomes

[0860] Experimental procedure:

[0861] (1). Take out the liver microsomes (20 mg protein / mL) from the -80 °C refrigerator, place them on a 37 °C water bath thermostatic oscillator for pre-warming and incubation for 3 min, and melt for use.

[0862] (2). Prepare a pre-incubation system mixed solution (without β-NADPH) according to the ratio of the "Composition of the Experimental Pre-incubation System" above.

[0863] (3). Prepare a 100 μM working solution of the test compound for standby.

[0864] (4). Control group (without β-NADPH): Take 25 μL of PB solution and add it to 75 μL of the incubation system mixture described in (2). Vortex for 30 s to mix well. The total reaction volume is 100 μL. Prepare duplicate samples. Place them in a 37 °C water bath constant temperature oscillator for incubation and start timing. The sampling time points are 0 min and 60 min.

[0865] (5). Sample group: Take 25 μL of β-NADPH solution (4 mM) and add it to 75 μL of the reaction system described in (2). Vortex for 30 s to mix well. The total reaction volume is 100 μL. Prepare duplicate samples. Place them in a 37 °C water bath constant temperature oscillator for incubation and start timing. The sampling time points are 0 min, 5 min, 15 min, 30 min, and 60 min.

[0866] (6). At each time point, take out the sample tube, add 300 μL of cold terminator (containing internal standard) to terminate the reaction.

[0867] (7). Vortex and centrifuge.

[0868] (8). Take 150 μL of the supernatant and add 150 μL of water. Vortex to mix well and perform LC-MS / MS injection analysis.

[0869] Data analysis

[0870] Calculate the half-life (t1 / 2) and clearance rate (CL) using the following first-order kinetic formula

[0871] Ct = C0 * e-kt

[0872] Ct = (1 / 2) * C0

[0873] t1 / 2 = ln2 / k = 0.693 / k

[0874] CL = Vd * k

[0875] Vd = 1 / Protein content in liver microsomes

[0876] CLint(liver) = CLint(mic) × Liver weight / body weight ratio × Microsomal protein concentration per gram of liver

[0877] The parameters in the formula are shown in Table 3:

[0878] Table 3 Parameter list

[0879]

[0880] The experimental results are shown in Table 4:

[0881] Table 4 Microsomal stability of compounds in different species

[0882]

[0883] In the human liver microsome stability experiment, the stability of compound 13 was significantly better than that of BI-3406; in the mouse liver microsome stability experiment, the stabilities of compounds 16 and 17 were significantly better than that of BI-3406. In summary, the stabilities of multiple compounds in different species of liver microsomes in the examples were significantly better than that of BI-3406.

[0884] Test Example 5 Rat Pharmacokinetic Properties of Compounds

[0885] The compound of Example 1 (10 mg / kg) was orally administered to SD rats (male, n = 3 / time point) fasted overnight. The rats were fed 2 h after dosing. Blood samples were collected before dosing and at 1.5 h, 4 h, and 8 h after dosing, and centrifuged at 4500 rpm for 10 min at 4 °C to obtain serum. The liver was collected before dosing and at 1.5 h, 4 h, and 8 h after dosing. Then the liver samples were quickly frozen and stored at -80 °C. 100 μL of MeOH / ACN (1:1, v / v) was added to 10 μL of serum to precipitate the mixture, followed by vortexing for 1 min and then centrifuging (11000 rpm) for 5 min to obtain the supernatant. 20 μL of the supernatant was dissolved in 20 μL of ACN / H2O (1:1, v / v) and analyzed by ultra-high performance liquid chromatography. 10 times the weight of MeOH / ACN (1:1, v / v) was added to the liver samples, and then homogenized with a homogenizer at 50 Hz for 120 s to obtain a homogenate. The homogenate was centrifuged (11000 rpm) for 5 minutes and the supernatant was collected. Then 20 μL of the supernatant was redissolved in 20 μL of ACN / H2O (1:1, v / v) and analyzed by ultra-high performance liquid chromatography. The results are shown in Table 5:

[0886] Table 5 Rat Pharmacokinetic Characteristics of Compounds

[0887]

[0888] As can be seen from Table 5, compound 1 had good plasma exposure and a good half-life in rats.

[0889] Test Example 6 In Vivo Antitumor Activity of Compounds

[0890] Human pancreatic cancer cells MIAPaCa-2 or PANC-1 (1.0×107 cells / mouse) were subcutaneously injected into the right side of female BALB / c mice (6 - 8 weeks old). The mice were administered orally with compound 1 (Cpd 1) (50 mg / kg), Trametinib (0.125 mg / kg, bid), and the combination of compound 1 and Trametinib (co-administration), twice a day (bid) for continuous administration. The mice were monitored daily and caliper measurements were started when the tumors became visible. The volume of the tumor was calculated using the following formula by measuring two perpendicular diameters: (L*W2) / 2, where L and W refer to the length and width of the tumor diameter. When the average tumor volume reached 100 mm3, the mice were grouped (D0, n = 5 / group) and administered the compounds. During the administration period, the tumor volume and the body weight of the mice were measured once every 3 days. The results are as follows Figures 2-5 as shown

[0891] Conclusion: The experimental results show that compound 1 of the embodiments of the present invention has good anti-tumor activity against different mutant tumor models of KRAS at single and combined doses. In the two anti-tumor models, compound 1 of the embodiments has no obvious effect on the body weight of mice at single and combined doses, showing good safety. Compound 1 of the embodiments has a certain inhibitory activity against the xenograft tumor of PANC-1 (KRAS G12D) nude mice, and the tumor growth inhibition rate is 40.6% (P < 0.01); compound 1 of the embodiments has a certain inhibitory activity against the xenograft tumor of MIAPaCa-2 (KRAS G12C) nude mice, and the tumor growth inhibition rate is 65.4% (P < 0.01). The tumor inhibition rate of the combination group with the MEK inhibitor Trametinib is 105.6%, and the tumors of all mice in the combination group regressed, and the activity was significantly enhanced compared with the single administration group (P < 0.001).

[0892] The applicant declares that the present invention uses the above embodiments to illustrate a polycyclic pyrimidine derivative as an SOS1 inhibitor, its preparation method and application, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the products of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

[0893] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0894] In addition, it should be noted that, in the above specific embodiments, the various specific technical features described can be combined in any suitable manner without conflict. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A polycyclic pyrimidine derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, characterized in that, Its structure is selected from the following structures, 、 。 2. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the polycyclic pyrimidine derivative as claimed in claim 1, its pharmaceutically acceptable salt, its tautomer or its stereoisomer, and a pharmaceutically acceptable carrier and / or excipient.

3. Use of the polycyclic pyrimidine derivative as claimed in claim 1, its pharmaceutically acceptable salt, its tautomer, its stereoisomer in the preparation of a medicament for preventing and treating cancer, wherein the cancer is selected from pancreatic cancer, leukemia, colorectal cancer, lung cancer, hepatocellular carcinoma, renal cancer, gastric cancer and cholangiocarcinoma.

4. Use of the polycyclic pyrimidine derivative as claimed in claim 1, its pharmaceutically acceptable salt, its tautomer, its stereoisomer alone or in combination with a MEK signaling pathway inhibitor in the preparation of a SOS1 inhibitor.

5. The use according to claim 4, wherein the MEK inhibitor is Trametinib.

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