Combination of pan-ras inhibitor and FAK inhibitor

Abstract

A combined drug combination, drug composition or combination product of a pan-RAS (ON) inhibitor compound and a FAK (focal adhesion kinase) inhibitor, and a method for using the combined drug combination, drug composition or combination product to prepare a drug for preventing or treating a related pathological condition.

Description

Combination of a pan-RAS inhibitor and a FAK inhibitor

[0001] Cross-reference to Related Applications

[0002] This application claims the benefit of and priority to the following Chinese patent application, which is hereby incorporated by reference in its entirety:

[0003] Patent Application No. 202411294000.7, filed with the National Intellectual Property Office on September 14, 2024. TECHNICAL FIELD

[0004] The present disclosure relates to a combination, a pharmaceutical composition or a combination product of a pan-RAS(ON) inhibitor compound and a FAK (Focal Adhesion Kinase, FAK) inhibitor, and a method for preventing or treating a related pathological disorder using the combination, the pharmaceutical composition or the combination product. BACKGROUND

[0005] RAS is a small GTPase responsible for signal switching, as a key component of RTK-mediated signaling pathways, involved in regulating cell growth and proliferation, cell differentiation, morphogenesis and apoptosis, etc. Under physiological conditions, RAS is mostly in the form of RAS(OFF) inactive state bound to GDP; after RAS mutation, the GTPase activity is impaired, making it more inclined to stay in the RAS(ON) active state bound to GTP, leading to continuous activation of RAS protein, and in turn continuously activating downstream signals such as MAPK and PI3K-AKT signaling pathways, resulting in uncontrolled cell proliferation and differentiation, forming tumors. RAS includes three subtypes of KRAS, NRAS and HRAS, and oncogenic mutations drive up to 30% of human cancers. Most oncogenic RAS mutations are gain-of-function missense changes at hotspots codons G12, G13 or Q61, among which KRAS mutations are the most common, accounting for 85%, commonly seen in non-small cell lung cancer (NSCLC), colorectal cancer (CRC) and pancreatic ductal adenocarcinoma (PDAC); NRAS mutations account for 12%, commonly seen in melanoma and acute myeloid leukemia; HRAS mutations account for 3%, commonly seen in bladder cancer and head and neck cancer. Given the key driving role of RAS in cancer, targeting RAS has become an important anti-tumor strategy.

[0006] Current KRAS inhibitors G12C Inhibitors AMG510 and MRTX849 have been approved for marketing successively for KRAS G12C mutant non-small cell lung cancer patients, but face problems such as low patient response rate and frequent drug resistance. In addition, such inhibitors target KRAS G12CThe (OFF) protein, while the GTP-binding KRAS (ON) protein is present in a larger proportion in tumors; these inhibitors target only KRAS. G12C This specific mutant, KRAS G12D KRAS G13 For other cancer mutation types such as KRAS, NRAS, and HRAS, effective targeted drugs are still lacking, limiting their clinical application. To address these issues, a class of pan-RAS(ON) inhibitors has emerged, such as the macrocyclic compounds developed by Revolution Medicines, Inc. These inhibitors form a ternary complex with the intracellular chaperone protein CYPA and the RAS(ON) protein, blocking the binding of RAS to downstream RAF, thus preventing signal transduction. These inhibitors can simultaneously target KRAS, NRAS, and HRAS mutations, expanding the range of indications. Furthermore, their mechanism of action—inhibiting RAS(ON) activity—exhibits superior tumor-suppressive effects. However, as the clinical development of pan-RAS(ON) inhibitors progresses, issues such as duration of response and drug resistance will become key factors affecting the efficacy of these drugs. Currently, there are reports that long-term use of pan-RAS inhibitors can lead to drug resistance, which can occur through activation of the YAP / TAZ / TEAD signaling pathway. This resistance may limit the long-term therapeutic efficacy of this therapy. Therefore, there is an urgent need to develop effective strategies to fully combat drug resistance problems that occur during the use of inhibitors, while improving the anti-tumor effects of pan-RAS(ON) inhibitors, and obtaining better and more effective clinical treatment drugs and regimens. Summary of the Invention

[0007] This disclosure provides a combination of drugs including pan-RAS(ON) inhibitors and FAK inhibitors.

[0008] In one aspect, this disclosure provides a method for treating a tumor disease in a subject in need, the method comprising administering a pan-RAS(ON) inhibitor and a FAK inhibitor to the subject thereby treating the subject.

[0009] Secondly, this disclosure also provides a pharmaceutical composition comprising a pan-RAS(ON) inhibitor and a FAK inhibitor, as well as pharmaceutically acceptable excipients.

[0010] Thirdly, this disclosure also provides a combination product comprising a first pharmaceutical composition and a second pharmaceutical composition, wherein the first pharmaceutical composition comprises at least one pan-RAS(ON) inhibitor and a pharmaceutically acceptable excipient, and the second pharmaceutical composition comprises at least one FAK inhibitor and a pharmaceutically acceptable excipient.

[0011] In another aspect, the present disclosure also provides a kit comprising a FAK inhibitor and instructions that the FAK inhibitor can be used in combination with a pan-RAS(ON) inhibitor for treating a tumor.

[0012] In another aspect, the present disclosure also provides a kit comprising a pan-RAS(ON) inhibitor and instructions that the RAS(ON) inhibitor can be used in combination with a FAK inhibitor for treating a tumor.

[0013] In some embodiments, the FAK inhibitor is selected from the group consisting of Ifebemtinib (IN-10018, BI-853520), Defactinib (VS-6063, PF-04554878), Narmafotinib (AMP-945), VS-4718 (PND-1186, SR-2516), TAE226, and GSK2256098, or a pharmaceutically acceptable salt thereof.

[0014] In some embodiments, the FAK inhibitor is selected from the group consisting of Ifebemtinib (IN-10018, BI-853520), Defactinib (VS-6063, PF-04554878), Narmafotinib (AMP-945), and VS-4718 (PND-1186, SR-2516), or a pharmaceutically acceptable salt thereof.

[0015] In some embodiments, the FAK inhibitor is selected from the group consisting of VS-4718 (PND-1186, SR-2516), TAE226, and GSK2256098, or a pharmaceutically acceptable salt thereof.

[0016] In some embodiments, the FAK inhibitor is VS-4718.

[0017] In some embodiments, the pan-RAS(ON) inhibitor is selected from any of the disclosed compounds in the following patent publications: WO2022060836A, WO2024067857A1, CN117534684A, CN117534687A, CN117534685A, WO2024060966A1, WO2024104364A1, WO2024169914A1, WO2024017859A1, WO2023025832A1, WO2024008834A1, WO2023232776A1, WO2025119392A1, WO2025087431A1, WO2024208934A1, WO2025045233A1, WO2024249299A2, WO2024222864A1, WO2024211712A, WO2024211663A, and WO2024153208A9, which are incorporated by reference in their entirety into the present disclosure.

[0018] In some embodiments, the pan-RAS(ON) inhibitor is selected from any of the disclosed compounds in the following patent publications: WO2022060836A, WO2024067857A1, CN117534684A, CN117534687A, CN117534685A, WO2024060966A1, WO2024104364A1, WO2024169914A1, WO2024017859A1, WO2023025832A1, WO2024008834A1, WO2023232776A1, and WO2024153208A9, which are incorporated by reference in their entirety into the present disclosure.

[0019] In some embodiments, the pan-RAS(ON) inhibitor is selected from the following compounds: compounds Al-Compound A608 disclosed in WO2022060836A, compounds Bl-Compound B25 disclosed in WO2022060836A, compounds Cl-Compound C21 disclosed in WO2022060836A, compounds prepared in Examples 1 to 16 of WO2024067857A1, compounds prepared in Examples 1 to 9 of CN117534684A, compounds prepared in Examples 1 to 9 of CN117534687A, compounds prepared in Examples 1 to 8 of CN117534685A, compounds prepared in Examples 1 to 61 of WO2024060966A1, compounds prepared in Examples 1 to 66 of WO2024104364A1, compounds prepared in Examples 1 to 127 of WO2024169914A1, compounds prepared in Examples 1 to 19 of WO2024017859A1, compounds prepared in Examples 1 to 26 of WO2023025832A1, compounds prepared in Examples 1 to 12 of WO2024008834A1, compounds prepared in Examples 1 to 58 of WO2023232776A1, compounds prepared in Examples 1 to 181 of WO2024153208A9, compounds prepared in Examples 1 to 4 of WO2025119392A1, compounds prepared in Examples 1 to 204 of WO2025087431A1, compounds prepared in Examples 1 to 47 of WO2024208934A1, compounds in Tables A-C of WO2025045233A1, compounds in Table 1 of WO2024249299A2, compounds prepared in Examples 1 to 3 of WO2024222864A, compounds in Table 1 of WO2024211712A1, and compounds in Table 1 of WO2024211663A1.

[0020] In some embodiments, the pan-RAS(ON) inhibitor is selected from the following compounds: compounds Al-Compound A608 disclosed in WO2022060836A, compounds Bl-Compound B25 disclosed in WO2022060836A, compounds Cl-Compound C21 disclosed in WO2022060836A, compounds prepared in Examples 1 to 16 of WO2024067857A1, compounds prepared in Examples 1 to 9 of CN117534684A, compounds prepared in Examples 1 to 9 of CN117534687A, compounds prepared in Examples 1 to 8 of CN117534685A, compounds prepared in Examples 1 to 61 of WO2024060966A1, compounds prepared in Examples 1 to 66 of WO2024104364A1, compounds prepared in Examples 1 to 127 of WO2024169914A1, compounds prepared in Examples 1 to 19 of WO2024017859A1, compounds prepared in Examples 1 to 26 of WO2023025832A1, compounds prepared in Examples 1 to 12 of WO2024008834A1, compounds prepared in Examples 1 to 58 of WO2023232776A1, and compounds prepared in Examples 1 to 181 of WO2024153208A9.

[0021] In some embodiments, the pan-RAS(ON) inhibitor is selected from any of the compounds disclosed in the following patent publications: WO2022060836A, WO2024067857A1, WO2024060966A1, WO2024104364A1, WO2024169914A1, and WO2024153208A9.

[0022] In some embodiments, the pan-RAS(ON) inhibitor is selected from the following compounds: compounds Al-Compound A608 disclosed in WO2022060836A, compounds Bl-Compound B25 disclosed in WO2022060836A, compounds Cl-Compound C21 disclosed in WO2022060836A, compounds prepared in Examples 1 to 16 of WO2024067857A1, compounds prepared in Examples 1 to 61 of WO2024060966A1, compounds prepared in Examples 1 to 66 of WO2024104364A1, compounds prepared in Examples 1 to 127 of WO2024169914A1, and compounds prepared in Examples 1 to 181 of WO2024153208A9.

[0023] In some embodiments, the pan-RAS(ON) inhibitor is selected from a compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0024] wherein,

[0025] is selected from

[0026] n is selected from 0, 1, 2, and 3;

[0027] A is selected from C3-C 12 cycloalkylene, 4-10 membered heterocyclylene, C6-C 10 arylene, and 5-12 membered heteroarylene, said C3-C 12 cycloalkylene, 4-10 membered heterocyclylene, C6-C 10 arylene, and 5-12 membered heteroarylene are optionally substituted with 1 or more R a ;

[0028] R 1 is selected from C(O)R 11 , C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl are optionally substituted with 1 or more R 1a ;

[0029] R 2 , R 3 , R 4 , R 7 , R 8 , and R 9 are independently selected from hydrogen, halogen, hydroxyl, thiol, amino, cyano, C1-C 10 alkyl, C1-C 10 alkoxy, C1-C 10 haloalkyl, and C3-C7cycloalkyl;

[0030] or, R 4 and R 7 , and the atom to which they are attached, together form a 4-10 membered heterocyclic ring, said 4-10 membered heterocyclic ring is optionally substituted with 1 or more Rb substituted;

[0031] R 5 is selected from hydrogen, halogen, hydroxyl, thiol, amino, cyano, C1-C4alkyl, C1-C4haloalkyl, and C1-C4alkoxy; 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, is optionally substituted with 1 or more R 5a substituted;

[0032] R 6 is selected from hydrogen, halogen, hydroxyl, thiol, amino, cyano, C1-C4alkyl, C1-C4haloalkyl, and C1-C4alkoxy;

[0033] R 11 is selected from amino, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said amino, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, is optionally substituted with 1 or more R 11a substituted;

[0034] each R a and R 10 is independently selected from halogen, amino, hydroxyl, thiol, cyano, and C1-C4alkyl;

[0035] each R 1a and R 11a is independently selected from halogen, amino, hydroxyl, thiol, cyano, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said amino, hydroxyl, thiol, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, is optionally substituted with 1 or more R 1aa substituted;

[0036] each R 1aa is independently selected from the group consisting of halogen, amino, hydroxyl, thiol, cyano, C1-C7alkyl, C1-C7haloalkyl, and C1-C7alkoxy;

[0037] each R 5a is independently selected from the group consisting of C1-C 10 alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkoxy, C3-C 12 cycloalkyl, 4-14 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said C1-C 10 alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkoxy, C3-C 12 cycloalkyl, 4-14 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl are optionally substituted with 1 or more R c ;

[0038] or, wherein one R 5a and R 4 , together with the atom to which they are attached, form an 8-10 membered heterocyclic ring, said 8-10 membered heterocyclic ring is optionally substituted with 1 or more R c ;

[0039] each R c is independently selected from the group consisting of halogen, amino, hydroxyl, thiol, cyano, C1-C7alkyl, C1-C7alkoxy, C3-C 10 cycloalkyl, and 4-12 membered heterocyclyl, said amino, hydroxyl, thiol, C1-C7alkyl, C1-C7alkoxy, C3-C 10 cycloalkyl, and 4-12 membered heterocyclyl are optionally substituted with 1 or more R h ;

[0040] R 5b and R 5b’ are independently selected from the group consisting of C1-C7alkyl, C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said C1-C7alkyl, C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl are optionally substituted with 1 or more R d ;

[0041] Or, R 5b and R 5b’ The atoms connected to it together form a 4-12 membered heterocyclic group, wherein the 4-12 membered heterocyclic group is optionally bounded by one or more R atoms. e replace;

[0042] Each R d and R e The radical is independently selected from halogen, hydroxyl, amino, C1-C7 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, and 4-10 membered heterocyclic groups, wherein the amino, C1-C7 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, and 4-10 membered heterocyclic groups are optionally surrounded by one or more R... f replace;

[0043] Each R f Independently selected from C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, and 4-10 membered heterocyclic groups, wherein the C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, and 4-10 membered heterocyclic groups are optionally surrounded by one or more R... g replace;

[0044] Each R b R g and R h Independently selected from halogens, hydroxyl groups, mercapto groups, amino groups, =O, C1-C4 alkyl groups, C1-C4 hydroxyalkyl groups, C1-C4 haloalkyl groups, (C1-C4 alkylene)OC1-C4 alkyl groups, and C1-C4 alkoxy groups;

[0045] One or more hydrogen atoms in the compound of formula (I), its stereoisomer, or its pharmaceutically acceptable salt may optionally be deuterium atoms.

[0046] In some embodiments, the compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt does not include (RMC-6236).

[0047] In some implementation schemes, Selected from

[0048] In some implementation schemes, Selected from

[0049] In some implementations, A is selected from C6-C. 10 arylene and 5-12-membered heteroarylene, the C6-C 10 The aryl and 5-12 heteroaryl groups are optionally enclosed by one or more R groups. a replace.

[0050] In some embodiments, A is selected from 5-12 membered heteroaryl, optionally substituted with 1 or more R a substituents.

[0051] In some embodiments, A is selected from 5-6 membered heteroaryl, optionally substituted with 1 or more R a substituents.

[0052] In some embodiments, A is selected from thiazole ring, optionally substituted with 1 or more R a substituents.

[0053] In some embodiments, A is selected from substituents. substituents. a substituents.

[0054] In some embodiments, R a is independently selected from halogen, amino, hydroxyl, thiol, and cyano.

[0055] In some embodiments, A is

[0056] In some embodiments, A is wherein * represents the attachment point to the indole ring.

[0057] In some embodiments, R 1 is selected from C(O)R 11 , C1-C 10 alkyl, C3-C 12 cycloalkyl, and 4-10 membered heterocyclyl, said C1-C 10 alkyl, C3-C 12 cycloalkyl, and 4-10 membered heterocyclyl optionally substituted with 1 or more R 1a substituents.

[0058] In some embodiments, R 1 is selected from C(O)R 11 , C1-C 10 alkyl, C4-C 12 cycloalkyl, and 4-10 membered heterocyclyl, said C1-C 10 alkyl, C4-C 12 cycloalkyl, and 4-10 membered heterocyclyl optionally substituted with 1 or more R 1a substituents.

[0059] In some embodiments, R 1 is selected from C(O)R 11, C1-C5alkyl, C3-C5cycloalkyl, and 4-6 membered heterocyclyl, said C1-C5alkyl, C3-C5cycloalkyl, and 4-6 membered heterocyclyl optionally substituted with 1 or more R 1a substituted.

[0060] In some embodiments, R 1 is selected from C(O)R 11 , C1-C5alkyl, C4-C5cycloalkyl, and 4-6 membered heterocyclyl, said C1-C5alkyl, C4-C5cycloalkyl, and 4-6 membered heterocyclyl optionally substituted with 1 or more R 1a substituted.

[0061] In some embodiments, R 1 is selected from C(O)R 11 , methyl, pentanyl, cyclopropyl, and tetrahydropyrrolyl, said methyl, pentanyl, cyclopropyl, and tetrahydropyrrolyl optionally substituted with 1 or more R 1a substituted.

[0062] In some embodiments, R 1 is selected from C(O)R 11 , methyl, pentanyl, and tetrahydropyrrolyl, said methyl, pentanyl, and tetrahydropyrrolyl optionally substituted with 1 or more R 1a substituted.

[0063] In some embodiments, each R 1a is independently selected from halogen, C1-C4alkyl, and C3-C4cycloalkyl, said C1-C4alkyl and C3-C4cycloalkyl optionally substituted with 1 or more R 10 ; 12 10 12 1aa substituted;

[0064] In some embodiments, each R 1a is independently selected from halogen, C1-C4alkyl, and C3-C4cycloalkyl, said C1-C4alkyl and C3-C4cycloalkyl optionally substituted with 1 or more R 1aa ;

[0065] In some embodiments, each R 1a is independently selected from halogen, C1-C4alkyl, and C3-C4cycloalkyl, said C1-C4alkyl and C3-C4cycloalkyl optionally substituted with 1 or more R 1aa substituted.

[0066] In some embodiments, R 11 is selected from 4-10 membered heterocyclyl, said 4-10 membered heterocyclyl optionally substituted with 1 or more R 11a substituted.

[0067] In some embodiments, R​​​11 Selected from 4-6 membered heterocyclic groups, wherein the 4-6 membered heterocyclic group is optionally coupled with one or more R... 11a replace.

[0068] In some implementation schemes, R 11 Selected from tetrahydropyrrole, wherein the tetrahydropyrrole group is optionally surrounded by one or more R... 11a replace.

[0069] In some implementation schemes, each R 11a Independently selected from halogen, amino, hydroxyl, mercapto, cyano and C1-C 10 Alkyl groups, said amino, hydroxyl, mercapto groups and C1-C 10 Alkyl groups are optionally surrounded by one or more R 1aa replace.

[0070] In some implementation schemes, each R 11a Independently selected from C1-C 10 Alkyl, the C1-C 10 Alkyl groups are optionally surrounded by one or more R 1aa replace.

[0071] In some implementation schemes, each R 11a Independently selected from C1-C4 alkyl groups, wherein the C1-C4 alkyl group is optionally surrounded by one or more R... 1aa replace.

[0072] In some implementation schemes, each R 11a The methyl group is independently selected from methyl groups, said methyl group optionally being surrounded by one or more R groups. 1aa replace.

[0073] In some implementation schemes, each R 1aa It is independently selected from halogens, C1-C7 alkyl groups, C1-C7 haloalkyl groups, and C1-C7 alkoxy groups.

[0074] In some implementation schemes, each R 1aa It is independently selected from halogens and C1-C4 alkyl groups.

[0075] In some implementation schemes, each R 1aa It is independently selected from methyl, chlorine and fluorine.

[0076] In some implementation schemes, R 2 R 3 Independently selected from hydrogen, halogen, hydroxyl, cyano and C1-C 10 alkyl.

[0077] In some implementation schemes, R 2 R 3 It is independently selected from C1-C4 alkyl groups, such as methyl.

[0078] In some implementation schemes, R 2 R 3 All are methyl groups.

[0079] In some implementation schemes, R 4 Selected from hydrogen, halogen, hydroxyl, cyano, C1-C 10 Alkyl and C1-C 10 Halogenated alkyl groups.

[0080] In some implementation schemes, R 4 Selected from C1-C4 alkyl and C1-C4 haloalkyl.

[0081] In some implementation schemes, R 4 Selected from ethyl and halogen-substituted ethyl groups.

[0082] In some implementation schemes, R 4 Selected from ethyl and fluorinated ethyl groups, such as trifluoroethyl.

[0083] In some implementation schemes, R 5 Selected from C6-C 10 Aryl and 5-10 heteroaryl, the C6-C 10 Aryl and 5-10 heteroaryl groups are optionally bounded by one or more R groups. 5a replace.

[0084] In some implementation schemes, R 5 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group is optionally surrounded by one or more R groups. 5a replace.

[0085] In some implementation schemes, R 5 Selected from pyridinyl, wherein the N in the pyridinyl group is optionally oxidized. The pyridinyl group is optionally surrounded by one or more R groups. 5a replace.

[0086] In some implementation schemes, R 5 for The Optionally by one or more R 5a replace.

[0087] In some implementation schemes, R 5 for N can be oxidized to form The Optionally by one or more R 5a replace.

[0088] In some implementation schemes, each R 5aindependently selected from the group consisting of C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R c C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R

[0089] In some embodiments, each R 5a is independently selected from the group consisting of C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R c C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R

[0090] In some embodiments, each R 5a is independently selected from the group consisting of methyl, ethyl, methoxy, cyclopropyl, morpholinyl, azetidinyl, oxetanyl, methyl, ethyl, methoxy, cyclopropyl, morpholinyl, azetidinyl, oxetanyl, optionally substituted with 1 or more R c C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, said C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl being optionally substituted with 1 or more R

[0091] In some embodiments, each R c is independently selected from the group consisting of amino, C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl, said amino, C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl, said amino, C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl being optionally substituted with 1 or more R 10 C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl, said amino, C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl being optionally substituted with 1 or more R h C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl, said amino, C1-C7alkyl, C1-C7alkoxy, C3-C8cycloalkyl, and 4-12 membered heterocyclyl being optionally substituted with 1 or more R

[0092] In some embodiments, each R c is independently selected from the group consisting of amino, C1-C4alkyl, C1-C4alkoxy, C3-C8cycloalkyl, and 4-8 membered heterocyclyl, said amino, C1-C4alkyl, C1-C4alkoxy, C3-C8cycloalkyl, and 4-8 membered heterocyclyl being optionally substituted with 1 or more R h C1-C4alkyl, C1-C4alkoxy, C3-C8cycloalkyl, and 4-8 membered heterocyclyl, said amino, C1-C4alkyl, C1-C4alkoxy, C3-C8cycloalkyl, and 4-8 membered heterocyclyl being optionally substituted with 1 or more R

[0093] In some embodiments, each R c is independently selected from the group consisting of amino, methyl, ethyl, methoxy, cyclopropyl, morpholinyl, azetidinyl, oxetanyl, and piperidinyl, said amino, methyl, ethyl, methoxy, cyclopropyl, morpholinyl, azetidinyl, oxetanyl, and piperidinyl are optionally substituted with 1 or more R h substituents.

[0094] In some embodiments, each R h is independently selected from halogen, hydroxyl, =0, C1-C4alkyl, C1-C4alkyleneOC1-C4alkyl, and C1-C4hydroxyalkyl.

[0095] In some embodiments, each R h is independently selected from fluorine, hydroxyl, =0, hydroxymethyl, methoxyethyl, and methyl.

[0096] In some embodiments, R 5b and R 5b’ together with the atom to which they are attached form a 4-6 membered heterocyclyl, said 4-6 membered heterocyclyl optionally substituted with 1 or more R e substituents.

[0097] In some embodiments, R 5b and R 5b’ together with the atom to which they are attached form a thietanyl and thiomorpholinyl, said thietanyl and thiomorpholinyl optionally substituted with 1 or more R e substituents.

[0098] In some embodiments, is selected from said is optionally substituted with 1 or more R e substituents.

[0099] In some embodiments, each R e is independently selected from halogen, hydroxyl, amino, C1-C7alkyl, and 4-10 membered heterocyclyl, said hydroxyl, amino, C1-C7alkyl, and 4-10 membered heterocyclyl optionally substituted with 1 or more R f substituents.

[0100] In some embodiments, each R e is independently selected from C1-C4alkyl and 4-6 membered heterocyclyl, said C1-C4alkyl and 4-6 membered heterocyclyl optionally substituted with 1 or more R f substituents.

[0101] In some embodiments, each R e is independently selected from ethyl and morpholinyl, said ethyl and morpholinyl optionally substituted with 1 or more R f substituents.

[0102] In some embodiments, each R f is independently selected from C1-C4alkoxy.

[0103] In some embodiments, R f is methoxy.

[0104] In some embodiments, each R 5a is independently selected from isopropyl, morpholinyl,

[0105] In some embodiments, each R 5a is independently selected from isopropyl,

[0106] In some embodiments, R 1 is C1-C4alkyl substituted cyclopropyl, and R 5a is selected from isopropyl,

[0107] In some embodiments, R 1 is C1-C4alkyl substituted cyclopropyl, and R 5a is selected from isopropyl,

[0108] In some embodiments, the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, has two R 5a .

[0109] In some embodiments, the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, has two R 5a , wherein one R 5a and R 4 , together with the atoms to which they are attached, collectively comprise a 9-membered heterocyclic ring, optionally substituted with one or more R c .

[0110] In some embodiments, the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, has two R 5a , wherein one R 5a and R 4 , together with the atoms to which they are attached, collectively comprise

[0111] In some embodiments, is selected from wherein R c is preferably methyl.

[0112] In some embodiments, R6 selected from hydrogen, halogen, hydroxyl, thiol, Ci-C4alkyl, and Ci-C4alkoxy.

[0113] In some embodiments, R 6 is selected from hydrogen and Ci-C4alkoxy.

[0114] In some embodiments, R 6 is hydrogen and ethoxy.

[0115] In some embodiments, R 7 is selected from hydrogen, halogen, hydroxyl, and cyano.

[0116] In some embodiments, R 7 is hydrogen.

[0117] In some embodiments, R 4 and R 7 together with the atom to which they are attached form a 6-7 membered heterocyclic ring, which is optionally substituted with 1 or more R b .

[0118] In some embodiments, each R b is independently selected from halogen, amino, hydroxyl, thiol, and cyano.

[0119] In some embodiments, each R b is independently selected from halogen, such as fluorine.

[0120] In some embodiments, is selected from said t is selected from 0, 1, 2, and 3.

[0121] In some embodiments, is selected from said t is selected from 0, 1, 2, and 3.

[0122] In some embodiments, is selected from

[0123] In some embodiments, R 8 is selected from hydrogen, halogen, hydroxyl, cyano, Ci-C 10 alkyl, and C3-C7cycloalkyl.

[0124] In some embodiments, R 8 is selected from hydrogen, cyano, Ci-C4alkyl, and C3-C7cycloalkyl.

[0125] In some embodiments, R 8 is hydrogen.

[0126] In some embodiments, R9 selected from hydrogen, halogen and hydroxyl.

[0127] In some embodiments, R 9 selected from hydrogen and halogen.

[0128] In some embodiments, R 9 selected from hydrogen and fluorine.

[0129] In some embodiments, the compound of formula (I) of the present disclosure, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of formula (I-1), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof,

[0130] wherein, R 1 , R 2 , R 3 , R 4 , R 5a , R 6 , R 7 , R 8 , R 9 , R 10 and n are as defined above.

[0131] In some embodiments, the compound of formula (I) of the present disclosure, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of formula (I-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof,

[0132] wherein, R 1 , R 2 , R 3 , R 4 , R 5a , R 6 , R 7 , R 8 and R 9 are as defined above.

[0133] In some embodiments, the compound of formula (I) of the present disclosure, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof,

[0134] (RMC-6236), (RMC-7977), (hereinafter referred to as Compound PA-1), (hereinafter referred to as Compound PA-2), (hereinafter referred to as Compound PA-3), (hereinafter referred to as Compound PA-4), (hereinafter referred to as Compound PA-5), (hereinafter referred to as Compound PA-6)

[0135] In some embodiments, the pan-RAS(ON) inhibitor is selected from RMC-6236, RMC-7977, Compound PA-1, Compound PA-2, Compound PA-3, Compound PA-4, Compound PA-5, and Compound PA-6, or a pharmaceutically acceptable salt thereof.

[0136] In some embodiments, the pan-RAS(ON) inhibitor is selected from RMC-6236, RMC-7977, Compound PA-1, Compound PA-2, Compound PA-3, Compound PA-4, and Compound PA-5, or a pharmaceutically acceptable salt thereof.

[0137] In some embodiments, the pan-RAS(ON) inhibitor is selected from RMC-7977, Compound PA-1, Compound PA-2, Compound PA-3, Compound PA-4, and Compound PA-5, or a pharmaceutically acceptable salt thereof.

[0138] In some embodiments, the pan-RAS(ON) inhibitor is Compound PA-1.

[0139] In a fourth aspect, the present disclosure also provides a method for treating a tumor disease, comprising administering to an individual in need of the treatment a therapeutically effective amount of any of the aforementioned combination drug, pharmaceutical composition or combination product.

[0140] In another aspect, the present disclosure also provides use of any of the aforementioned combination drug, pharmaceutical composition or combination product in the manufacture of a medicament for treating a tumor disease.

[0141] In another aspect, the present disclosure also provides use of any of the aforementioned combination drug, pharmaceutical composition or combination product in treating a tumor disease.

[0142] In some embodiments, the tumor disease of the present disclosure includes, but is not limited to, lung cancer, colorectal cancer (such as colon cancer), pancreatic cancer, endometrial cancer, myeloma, breast cancer, ovarian cancer, gastric cancer, esophageal cancer, prostate cancer, cervical cancer, cholangiocarcinoma, or uterine carcinosarcoma.

[0143] In some embodiments, the tumor disease of the present disclosure is selected from pancreatic cancer, colorectal cancer, lung cancer, endometrial cancer, and myeloma.

[0144] In some embodiments, the lung cancer is preferably non-small cell lung cancer.

[0145] In alternative embodiments, the pan-RAS(ON) inhibitor and the FAK inhibitor in the combination can be packaged separately or together.

[0146] In some embodiments, the pan-RAS(ON) inhibitor and the FAK inhibitor in the use or method of treatment are each in the form of a pharmaceutical composition, which can be administered simultaneously, sequentially or at intervals.

[0147] In some embodiments, the pan-RAS(ON) inhibitor and the FAK inhibitor in the use or method of treatment are each administered in the form of administration at intervals.

[0148] In some embodiments, the pan-RAS(ON) inhibitor and the FAK inhibitor in the use or method of treatment are each administered in the form of administration at intervals.

[0149] In some embodiments, the pan-RAS(ON) inhibitor and the FAK inhibitor in the use or method of treatment are each administered in the form of administration at intervals.

[0150] In some embodiments, the FAK inhibitor is administered at a dose ranging from 5 mg / day to 1500 mg / day in an adult. In a particular embodiment, Ifebemtinib or a pharmaceutically acceptable salt thereof is administered at a dose ranging from 5 mg / day to 100 mg / day in an adult, for example, Ifebemtinib or a pharmaceutically acceptable salt thereof is administered at a dose ranging from 25 mg / day to 100 mg / day in an adult, the dose being calculated in free form.

[0151] In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered orally (PO) at about 100 mg to about 800 mg, twice a day (BID), e.g., about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 100 mg to about 400 mg, about 100 mg to about 200 mg, about 200 mg to about 1000 mg, about 400 mg to about 1000 mg, about 600 mg to about 1000 mg, about 800 mg to about 1000 mg, about 200 mg to about 800 mg, about 200 mg to about 600 mg, about 200 mg to about 400 mg, about 400 mg to about 800 mg, or about 400 mg to about 600 mg, PO, BID. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered at about 200 mg to about 400 mg, PO, BID. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered at about 100 mg, PO, BID. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered at about 200 mg, PO, BID. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered at about 300 mg, PO, BID. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered at about 400 mg, PO, BID. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered at about 600 mg. In some embodiments, the FAK inhibitor (e.g., Defactinib) is administered orally.

[0152] In some embodiments, the FAK inhibitor can be administered in the use or method of treatment with a frequency of 3 times daily (t.i.d), 2 times daily (b.i.d), or 1 time daily (q.d).

[0153] In some embodiments, the pan-RAS(ON) inhibitor can be administered in the use or method of treatment with a frequency of 3 times daily (t.i.d), 2 times daily (b.i.d), or 1 time daily (q.d); in a dosage range of 1 mg / day - 1000 mg / day in an adult, preferably 10 mg / day - 400 mg / day, more preferably 80 mg / day - 300 mg / day.

[0154] Technical effects:

[0155] The administration of the combination, pharmaceutical composition or combination product of the present disclosure comprising a pan-RAS(ON) inhibitor and a FAK inhibitor helps to:

[0156] (1) produce a better therapeutic effect in reducing the growth of a tumor or even eliminating a tumor, or provide a smaller amount of administration, as compared to the administration of a pan-RAS(ON) inhibitor or a FAK inhibitor alone; and / or,

[0157] (2) the patient has better tolerability with fewer adverse effects and / or complications compared to administration of a pan-RAS(ON) inhibitor or a FAK inhibitor alone; and / or,

[0158] (3) there is no report of a pan-RAS(ON) inhibitor or a FAK inhibitor combination or pharmaceutical composition that reduces the drug resistance generated by long-term administration of the patient compared to the prior art (a pan-RAS(ON) inhibitor alone); and / or,

[0159] (4) the combination drug, the pharmaceutical composition or the combination product has an inhibitory effect on a variety of RAS-dependent tumor cells, and such a broad application range can treat tumor diseases carrying more types of RAS mutations.

[0160] Definitions

[0161] Unless otherwise indicated, the terms used in the present disclosure have the following meanings, and the definitions of groups and terms described in the present disclosure, including the definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in examples, etc., can be combined and integrated with each other arbitrarily. A particular term should not be considered indefinite or unclear without a specific definition, but should be understood according to the ordinary meaning in the art. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof.

[0162] Herein represents a point of attachment.

[0163] Certain compounds of the present application can exist in atropisomeric forms, which are conformational isomers that arise when rotation about a single bond in the molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule. The compounds of the present disclosure include all atropisomers, either as pure individual atropisomers, or as atropisomers enriched in one, or as non-specific mixtures of each. Isomers can be separated if the potential energy for rotation about the single bond is sufficiently high and the interconversion between conformations is sufficiently slow. For example, represents a pair of atropisomers in which the represents a pair of atropisomers in which the represents that the side stereo orientation is outward, represents that the side stereo orientation is inward.

[0164] The graphical representation of racemates or enantiomerically pure compounds herein is from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, the wedge solid bond and the wedge dashed bond The absolute configuration of a solid center is represented by direct real keys and direct virtual keys. It indicates the relative configuration of a stereocenter (such as the cis-trans configuration of alicyclic compounds).

[0165] When one of the variables is selected as a chemical bond or does not exist, it means that the two groups it is connected to are directly connected. For example, when L in ALZ represents a bond, it means that the structure is actually AZ.

[0166] If the linking group mentioned in this article does not specify its linking direction, then its linking direction is arbitrary. For example, when the structural unit... L in 1 When selected from "C1-C3 alkylene-O", L 1 Both loops Q and R can be connected in a left-to-right direction. 1 Composed of "cyclo-Q-C1-C3 alkylene-OR" 1 Alternatively, rings Q and R can be connected from right to left. 1 Composed of "cyclo-QO-C1-C3 alkylene-R" 1 ".

[0167] When a substituent is cross-bonded to two atoms on a ring, it can bond to any atom on that ring. For example, structural units. R represents 10 Substitution can occur at any position on the ring.

[0168] The compounds disclosed herein may have asymmetric atoms such as carbon, sulfur, nitrogen, and phosphorus atoms, or asymmetric double bonds, and therefore may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E- and Z-type geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof or other mixtures, such as mixtures enriched with enantiomers or diastereomers. All such isomers and mixtures thereof are within the scope of the definition of the compounds disclosed herein. Alkyl groups or other substituents may contain additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms. All such isomers involved in all substituents, and mixtures thereof, are also included within the scope of the definition of the compounds disclosed herein. The compounds containing asymmetric atoms disclosed herein can be isolated in optically active pure form or in racemic form. The optically active pure form can be separated from racemic mixtures or synthesized using chiral starting materials or chiral reagents.

[0169] The term "substituted" means that any one or more hydrogen atoms on the designated atom is replaced with a substituent group, provided that the valency of the designated atom is not exceeded and that the substituted compound is stable. When the substituent is oxo (i.e., =0), it is meant that two hydrogen atoms are replaced, and oxo will not occur on an aromatic group.

[0170] The terms "optionally," "optionally," "optionally" or "optionally" mean that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, ethyl "optionally" substituted with one or more halogens means that the ethyl group can be unsubstituted (CH2CH3), mono-substituted (CH2CH2F, CH2CH2C1, etc.), poly-substituted (CHFCH2F, CH2CHF2, CHFCH2C1, CH2CHC12, etc.), or fully substituted (CF2CF3, CF2CC13, CC12CC13, etc.). One skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern will be introduced that is not spatially possible and / or synthetically feasible.

[0171] When any variable (e.g., R a , R b ) occurs more than one time in a compound, its definition in each instance is independent of the other occurrences. For example, if a group is substituted with 2 R b , then each R b has independent options.

[0172] C m -C n herein means an integer number of carbon atoms in the range m-n. For example "C1-C 10 " means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0173] The term "alkyl" means a hydrocarbon group of formula C n H 2n+1 which can be straight chained or branched. The term "C1-C 10"Alkyl" can be understood as representing a straight-chain or branched saturated hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1 The term "C1-C7 alkyl" can be understood as referring to alkyl groups having 1 to 7 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C5 alkyl" can be understood as referring to straight-chain or branched saturated alkyl groups having 1 to 5 carbon atoms. The term "C1-C4 alkyl" can be understood as referring to straight-chain or branched saturated alkyl groups having 1 to 4 carbon atoms. The term "C1-C3 alkyl" can be understood as referring to straight-chain or branched saturated alkyl groups having 1 to 3 carbon atoms. The term "C5-C6 alkyl" can also be understood as referring to straight-chain or branched saturated alkyl groups having 1 to 3 carbon atoms. 10 "Alkyl" can be understood as referring to a straight-chain or branched saturated alkyl group having 5 to 10 carbon atoms. The "C1-C" 10 "alkyl" can include "C1-C6 alkyl", "C1-C4 alkyl", "C1-C3 alkyl" or "C5-C6 alkyl". 10 The term "alkyl" is used within the range of "C1-C6 alkyl," which may further include "C1-C4 alkyl" or "C1-C3 alkyl." The term "halogenated alkyl" is intended to include both monohalogenated and polyhalogenated alkyl groups. For example, the term "C1-C6 alkyl" may include "C1-C4 alkyl" or "C1-C3 alkyl." 10 "Haloalkyl" refers to a C1-C alkyl group as defined above that has been substituted with one or more halogens. 10 Alkyl groups include, but are not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl. The term "hydroxyalkyl" is intended to include both monohydroxy-substituted and polyhydroxy-substituted alkyl groups. For example, the term "C1-C4 hydroxyalkyl" refers to a C1-C4 alkyl group as defined above that is substituted with one or more hydroxyl groups.

[0174] The term "alkoxy" refers to a group formed by the loss of a hydrogen atom from a hydroxyl group in straight-chain or branched alcohols; it can be understood as "alkyloxy" or "alkyl-O-". The term "C1-C"... 10 "Alkoxy" can be understood as "C1-C" 10 "alkyloxy" or "C1-C" 10alkyl-O-". The term "C1-C7alkoxy" can be understood as "C1-C7alkyl oxy" or "C1-C7alkyl-O-". Said "C1-C7alkoxy" can further comprise "C1-C3alkoxy". 10 alkoxy" can comprise "C1-C7alkoxy" and "C1-C3alkoxy" and the like, said "C1-C7alkoxy" can further comprise "C1-C3alkoxy".

[0175] The term "alkenyl" refers to a straight-chain or branched unsaturated aliphatic hydrocarbon group consisting of carbon and hydrogen atoms and having at least one double bond. The term "C2-C 10 alkenyl" can be understood as denoting a straight-chain or branched unsaturated hydrocarbon group comprising one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, the term "C6-C 10 alkenyl" can be understood as denoting a straight-chain or branched unsaturated hydrocarbon group comprising one or more double bonds and having 6, 7, 8, 9 or 10 carbon atoms, the term "C2-C 10 alkenyl" can comprise "C2-C6alkenyl", "C2-C4alkenyl", "C6-C 10 alkenyl", C2 or C3 alkenyl. It can be understood that in case said alkenyl group comprises more than one double bond, said double bonds can be separated from each other or conjugated. Particular examples of said alkenyl group include, but are not limited to, ethenyl, allyl, (E)-2-methylethenyl, (Z)-2-methylethenyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl or (Z)-1-methylprop-1-enyl and the like.

[0176] The term "alkynyl" refers to a straight-chain or branched unsaturated aliphatic hydrocarbon group consisting of carbon and hydrogen atoms and having at least one triple bond. The term "C2-C 10 alkynyl" can be understood as denoting a straight-chain or branched unsaturated hydrocarbon group comprising one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The term "C2-C 10 Examples of "alkynyl" include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡CCH 3、 -CH2C≡CH), but-1-ynyl, but-2-ynyl or but-3-ynyl. The term "C2-C 10 alkynyl" can comprise "C2-C3alkynyl", examples of "C2-C3alkynyl" include ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH3), prop-2-ynyl (-CH2C≡CH).

[0177] The term "cycloalkyl" refers to a fully saturated carbocyclic group that exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is typically a 3- to 20-membered ring. The term "C3-C" is also used. 12 "Cycloalkyl" refers to a cycloalkyl group having 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring carbon atoms. The term "C3-C6 cycloalkyl" refers to a cycloalkyl group having 3, 4, 5, or 6 ring carbon atoms. The term "cycloalkylene" is a residue derived from a cycloalkyl group by further removing a hydrogen atom.

[0178] The term "heterocyclyl" or "heterocycle" refers to a monocyclic, fused ring, spiro, or bridged ring radical, which is completely saturated or partially saturated (not an aromatic heteroaromatic overall), having from 1 to 5 (e.g., 1 to 3 or 1 to 2) heteroatoms or groups of heteroatoms (i.e., groups of atoms containing heteroatoms) in its ring atom count, including but not limited to nitrogen atoms (N), oxygen atoms (O), sulfur atoms (S), phosphorus atoms (P), boron atoms (B), -S(=0)2-, -S(=0)-, -P(=0)2-, -P(=0)-, -NH-, -S(=0)(=NH)-, -C(=0)NH-, or -NHC(=0)NH-, and the like, in its ring atom count. The term "4-10 membered heterocyclyl" refers to a heterocyclyl having a ring atom count of 4, 5, 6, 7, 8, 9, or 10, and having from 1 to 5 heteroatoms or groups of heteroatoms independently selected from those described above in its ring atom count. A "4-10 membered heterocyclyl" can include a "4-7 membered heterocyclyl." The term "4-7 membered heterocyclyl" refers to a heterocyclyl having a ring atom count of 4, 5, 6, or 7, and having from 1, 2, 3, 4, or 5 heteroatoms or groups of heteroatoms independently selected from those described above in its ring atom count. Specific examples of 4-membered heterocyclyls include, but are not limited to, azetidinyl or oxetanyl; specific examples of 5-membered heterocyclyls include, but are not limited to, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydro-lH-pyrrolyl; specific examples of 6-membered heterocyclyls include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl, or 4H-[l,3,4]thiadiazinyl; and specific examples of 7-membered heterocyclyls include, but are not limited to, diazepanyl. The heterocyclyl can also be a bicyclic radical, where specific examples of 5,5 membered bicyclic radicals include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(lH)-yl; specific examples of 5,6 membered bicyclic radicals include, but are not limited to, hexahydropyrrolo[l,2-a]pyrazin-2(lH)-yl, 5,6,7,8-tetrahydro-[l,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazo[l,5-a]pyrazinyl. Optionally, the heterocyclyl can be a benzo-fused ring of the aforementioned 4-7 membered heterocyclyl, specific examples include, but are not limited to, dihydroisoquinolinyl, and the like. A "4-10 membered heterocyclyl" can include a "5-10 membered heterocyclyl," a "4-7 membered heterocyclyl," a "5-6 membered heterocyclyl," a "6-8 membered heterocyclyl," a "4-10 membered heterocycloalkyl," a "5-10 membered heterocycloalkyl," a "4-7 membered heterocycloalkyl," a "5-6 membered heterocycloalkyl," a "6-8 membered heterocycloalkyl," and the like. A "4-7 membered heterocyclyl" can further include a "4-6 membered heterocyclyl," a "5-6 membered heterocyclyl," a "4-7 membered heterocycloalkyl," a "4-6 membered heterocycloalkyl," a "5-6 membered heterocycloalkyl," and the like.Although some bicyclic heterocyclic groups in this disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole remains non-aromatic. The term "subheterocyclic group" refers to a residue derived by further removing a hydrogen atom from a heterocyclic group.

[0179] The term "heterocyclic alkyl" refers to a fully saturated cyclic group existing in the form of a monocyclic, fused, bridged, or spirocyclic ring, wherein the ring atoms contain 1-5 heteroatoms or heteroatom groups (i.e., groups containing heteroatoms). These "heteroatoms or heteroatom groups" include, but are not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=O)2-, -S(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH-, or -NHC(=O)NH-. The term "4-10 membered heterocyclic alkyl" refers to a heterocyclic alkyl group with 4, 5, 6, 7, 8, 9, or 10 ring atoms, and its ring atoms contain 1-5 independently selected heteroatoms or heteroatom groups as described above. The term "5-10 membered heterocyclic alkyl" refers to a heterocyclic alkyl group with 5, 6, 7, 8, 9 or 10 ring atoms, and whose ring atoms contain 1 to 5 independent heteroatoms or heterogroups selected from those described above. "4-10-membered heterocyclic alkyl" and "5-10-membered heterocyclic alkyl" include "4-7-membered heterocyclic alkyl", wherein specific examples of 4-membered heterocyclic alkyl include, but are not limited to, acridine, oxadiazolyl, or thiobutylcycloyl; specific examples of 5-membered heterocyclic alkyl include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, imidazolyl, or tetrahydropyrazolyl; specific examples of 6-membered heterocyclic alkyl include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, morpholinyl, piperazine, 1,4-thiaoxalyl, 1,4-dioxane, thiomorpholinyl, 1,3-dithiaalkyl, or 1,4-dithiaalkyl; and specific examples of 7-membered heterocyclic alkyl include, but are not limited to, azirheptanyl, oxaheptanyl, or thioheptanyl.

[0180] The term "aryl" refers to an aromatic ring group consisting of an all-carbon monocyclic or fused polycyclic aromatic cyclic group with a conjugated π-electron system. Aryl groups can have 6-20, 6-14, or 6-12 carbon atoms. The term "C6-C"... 10 "Aryl" can be understood as an aryl group having 6 to 10 carbon atoms. The term "C6-C7 aryl" can be understood as an aryl group having 6 to 7 carbon atoms. For example, a ring with 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring with 9 carbon atoms ("C9 aryl"), such as indenyl or indenyl; or a ring with 10 carbon atoms ("C9 aryl"). 10 Aryl), such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl. The term "aryl" refers to a residue derived from an aryl group by further removing a hydrogen atom.

[0181] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system having aromaticity, which contains at least one ring atom selected from N, O, S, the remaining ring atoms being C. The term "5-12 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11 or 12 ring atoms, e.g. 5 or 6 or 9 or 10 or 11 or 12 ring atoms, and which contain 1-5, e.g. 1-3, heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and the like and their benzo derivatives, e.g. benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzoisoxazolyl, benzoimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and the like and their benzo derivatives, e.g. quinolinyl, quinazolinyl or isoquinolinyl and the like; or azocinyl, indolizinyl, purinyl and the like and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenoxazinyl, phenothiazinyl or phenoxazinyl and the like. The term "6-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 6, 7, 8, 9 or 10 ring atoms, e.g. 6 or 9 or 10 ring atoms, and which contain 1-5, e.g. 1-3, heteroatoms independently selected from N, O and S. The term "5-6 membered heteroaryl" refers to aromatic ring systems having 5 or 6 ring atoms, and which contain 1-3, e.g. 1-2, heteroatoms independently selected from N, O and S. The term "heteroarylene" is a residue derived from a heteroaryl group by further removal of one hydrogen.

[0182] The term "halo" or "halogen" refers to fluoro, chloro, bromo or iodo.

[0183] The term "hydroxy" refers to an -OH group.

[0184] The term "cyano" refers to a -CN group.

[0185] The term "amino" refers to an -NH2 group.

[0186] As used herein, the term "RAS(ON) inhibitor" refers to an inhibitor that targets, i.e., selectively binds to or inhibits, the active state of RAS bound to GTP (e.g., with selectivity over the inactive state of RAS bound to GDP). Inhibition of the active state of RAS bound to GTP includes, for example, inhibition of oncogenic signaling from the active state of RAS bound to GTP. In some embodiments, a RAS(ON) inhibitor is an inhibitor that selectively binds to and inhibits the active state of RAS bound to GTP.

[0187] As used herein, the term "pan-RAS(ON) inhibitor" refers to a RAS(ON) inhibitor of at least 3 RAS variants having missense mutations in one of the following positions: 12, 13, 59, 61, or 146, in some embodiments, the pan-RAS(ON) inhibitor refers to a RAS(ON) inhibitor of at least 3 RAS variants having missense mutations in one of the following positions: 12, 13, and 61.

[0188] The term "combination drug" refers to a combination of two or more active ingredients (administered as the active ingredient itself or in the form of a pharmaceutical composition thereof, or as a derivative, prodrug or a pharmaceutically acceptable salt or ester thereof, etc., of each active ingredient) administered simultaneously or sequentially.

[0189] The term "combination product" refers to a non-fixed combination, in which the active agent and at least one additional active agent can be administered simultaneously or within time intervals, particularly in cases where these time intervals allow the combination partners to exhibit a cooperative (e.g., synergistic) effect. The term "non-fixed combination" means that the active ingredients (e.g., one active agent and at least one additional active agent) are each administered to a patient as separate entities either simultaneously or sequentially within time intervals that do not allow for the interaction of the two compounds in the body in a way that would impair their therapeutic effect.

[0190] The term "pharmaceutically acceptable" pertains to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0191] The term "pharmaceutically acceptable salt" includes salts formed with free acids or salts formed with free bases, for example, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, etc. can be mentioned.

[0192] The doses of the compounds of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof involved in the present disclosure are calculated based on the molecular weight of the free form of the compound of formula (I), unless otherwise specified. The doses of other drug molecules involved in the present disclosure, such as RMC-6236, RMC-7977, Compound PA-1, Compound PA-2, Compound PA-3, Compound PA-4, Compound PA-5, Compound PA-6, and Ifebemtinib, Defactinib, Narmafotinib, PND-1186, TAE226 and GSK2256098, or a pharmaceutically acceptable salt thereof, are calculated based on the molecular weight of the free form, unless otherwise specified.

[0193] The term "pharmaceutical composition" refers to a mixture of the active ingredient with a pharmaceutically acceptable excipient which can be compounded as is, or further processed in some manner as is known in the art.

[0194] The pharmaceutical composition of the present disclosure can be administered in a suitable variety of routes, or the components in a combination drug or combination product can each independently be administered in a suitable variety of routes, typical routes including, but not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, subdermal, subcapsular, subarachnoid, intravenous, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraarticular, intraspinal, transtracheal, epidural, and sternal administration.

[0195] The term "pharmaceutically acceptable excipient" refers to those excipients that do not stimulate an undesirable response in an organism, and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0196] The term "combination" or "combined use" refers to two or more active substances that can each be administered simultaneously, or sequentially in any order, as unitary agents.

[0197] In the present context, the terms "subject", "patient" or "individual" are used interchangeably. In some embodiments, the term "subject", "patient" or "individual" is a mammal. In some embodiments, the subject, patient or individual is a mouse. In some embodiments, the subject, patient or individual is a human.

[0198] The components in the combination drug of the present disclosure can each independently be present in the form of a pharmaceutical composition.

[0199] The components in the combination drug of the present disclosure can each independently, or some or all of them collectively, be in a suitable dosage form, including but not limited to tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, and the like oral forms, or sterile solutions, suspensions, or lyophilized products, and the like parenteral administration forms.

[0200] The words "comprise" or "comprising" and variations such as "comprises" or "comprising", will be understood to imply a non-exclusive inclusion. Like terms have like definition.

[0201] The present disclosure also includes isotopically-labeled compounds of the present disclosure which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, iodine, and chlorine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 123 I, 125 I, and 36 Cl, and the like.

[0202] Certain isotopically-labeled compounds of the present disclosure (for example, those 3 H, and 14 C) can be used in compound and / or substrate tissue distribution analysis. Tritiated (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes such as 15 O, 13 N, 11 C, and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically-labeled compounds of the present disclosure can generally be prepared by

[0203] The pharmaceutical compositions of the present disclosure can be prepared by combining a compound of the present disclosure with a suitable pharmaceutically acceptable excipient, such as can be formulated into solid, semi-solid, liquid, or gaseous dosage forms, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres, aerosols, and the like.

[0204] Typical routes of administering a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0205] The pharmaceutical compositions of the present disclosure can be manufactured in a manner that is generally known, e.g., by means of conventional mixing, dissolving, granulating, emulsifying, levitating, lyophilizing processes, and the like method.

[0206] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by combining the active compounds with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, dragees, sugar-coated tablets, capsules, liquids, gels, slurries, suspensions, and the like, for oral administration to a patient.

[0207] Solid oral compositions can be prepared by conventional mixing or compaction methods. For example, the active compounds can be mixed with a solid excipient, optionally ground, and if necessary, with other suitable excipients, and then processed into granules, which are filled into tablets or sugar-coated tablets. Suitable excipients include, but are not limited to, binding agents, diluents, disintegrating agents, lubricants, glidants, or flavoring agents.

[0208] The pharmaceutical composition can also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

[0209] The term "treatment" generally refers to obtaining a desired pharmacologic and / or physiologic effect. The effect can be therapeutic in whole or in part to stabilize or cure a disease and / or side effects resulting from the disease. "Treatment" in the present disclosure encompasses any treatment of a patient's disease, including: (a) inhibiting the disease symptoms, i.e., arresting their development; or (b) relieving the disease symptoms, i.e., causing regression of the disease or symptoms.

[0210] The term "effective amount" means the amount of a compound of the present application which (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein. The amount of a compound of the present disclosure that will constitute an "effective amount" will vary depending on the compound, the disease state and its severity, the manner of administration, and the age of the mammal to be treated, but can be routinely determined by the skilled practitioner according to his own knowledge and the disclosure.

[0211] The term "administering" means physically introducing a composition comprising a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those of skill in the art.

[0212] The compounds disclosed herein can be prepared by a variety of synthetic methods known to those skilled in the art, such as the specific embodiments of RMC-6236 and RMC-7977 as described in WO2022060836A, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art; and the specific embodiments of compounds PA-1 and PA-2 as described in WO2024067857A1, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art.

[0213] The chemical reactions in the specific embodiments of this disclosure are carried out in a suitable solvent, which must be suitable for the chemical changes of this disclosure and the reagents and materials required therefor. In order to obtain the compounds of this disclosure, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0214] Abbreviations:

[0215] EA represents ethyl acetate; TBDPS represents tert-butyldiphenylsilyl; TBDPSCl represents tert-butyldiphenylchlorosilane; DCM represents dichloromethane; DMF represents N,N-dimethylformamide; THF represents tetrahydrofuran; MeOH represents methanol; TsOH .H20 represents p-toluenesulfonic acid monohydrate; TsCl represents p-toluenesulfonyl chloride; n-BuLi represents n-butyllithium; Boc20 represents di-tert-butyl dicarbonate; TFA: trifluoroacetic acid; DIEA or DIPEA represents N,N-diisopropylethylamine; Pd(dppf)Cl2 represents [1,1 '-bis(diphenylphosphino) ferrocene]dichloropalladium(II); Pd(dtbpf)Cl2 represents 1,1 '-bis(di-tert-butylphosphino) ferrocene dichloropalladium; HATU represents O-(7-azabenzotriazol-1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate); Et3N or TEA represents triethylamine; PPh3: triphenylphosphine; Ru-L(S,S) represents (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethane diamine (p-isopropylbenzene) chloro ruthenium; [Ir(cod)Cl]2 represents 1,5-cyclooctadiene iridium chloride dimer; B2Pin2 represents bis(pinacolato)diboron or 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane); COMU represents (2-oximino-cyanoacetic acid ethyl ester)-N,N-dimethyl-morpholino urea hexafluorophosphate; ACN represents acetonitrile; NIS represents N-iodosuccinimide; KOAc represents potassium acetate; DME represents ethylene glycol dimethyl ether; EtI represents iodoethane; EDCI represents 1 -ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; HOBT represents 1 -hydroxybenzotriazole; AcOH represents acetic acid; Boc represents tert-butoxycarbonyl; Cbz represents carbobenzyloxy; toluene represents methylbenzene; dioxane represents dioxane; TMSCHN2 represents trimethylsilyldiazomethane; Pd(PPh3)2Cl2 represents bis(triphenylphosphine)palladium dichloride; Ac20 represents acetic anhydride; TCFH represents N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate; NMI represents N-methylimidazole; Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium; SPhos represents 2-dicyclohexylphosphino-2',6'-dimethoxy-biphenyl; IPA / i-PrOH represents isopropyl alcohol; DMAP represents 4-dimethylaminopyridine; NMP represents N-methylpyrrolidone; LDA represents lithium diisopropylamide; DTBA represents di-tert-butyl azodicarboxylate; DMPUN represents N,N-dimethylpropynyl urea; TBAF represents tetrabutylammonium fluoride; DMSO represents dimethyl sulfoxide; NMM represents N-methylmorpholine; (CH2O) n represents paraformaldehyde; LC-MS represents liquid chromatography-mass spectrometry; MS represents mass spectrometry; 1HNMR stands for nuclear magnetic resonance hydrogen spectrum; ESI stands for electrospray ionization; DTT stands for dithiothreitol; HEPES stands for 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid; PBS stands for phosphate buffered saline; BSA stands for bovine serum albumin; IC 50 IC50 stands for half maximal inhibitory concentration, the concentration of a substance which induces a response halfway between the baseline and maximum achieved. BRIEF DESCRIPTION OF DRAWINGS

[0216] Figure 1 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS G12C mutant human pancreatic cancer MiaPaCa-2 cell proliferation inhibition.

[0217] Figure 2 shows a matrix plot of synergistic effect of compound RMC-6236 with co-drug Ifebemtinib (top left), compound RMC-6236 with co-drug VS-4718 (top right), compound RMC-6236 with co-drug Defactinib (bottom left) and compound RMC-6236 with co-drug Narmafotinib (bottom right) on KRAS G12D mutant human colon cancer GP2d cell proliferation inhibition.

[0218] Figure 3 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS G12D mutant human colon cancer GP2d cell proliferation inhibition.

[0219] Figure 4 shows a matrix plot of synergistic effect of compound RMC-6236 with co-drug Ifebemtinib (top left), compound RMC-6236 with co-drug VS-4718 (top right), compound RMC-6236 with co-drug Defactinib (bottom left) and compound RMC-6236 with co-drug Narmafotinib (bottom right) on KRAS G12D mutant human pancreatic cancer AsPC-1 cell proliferation inhibition.

[0220] Figure 5 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS G12D Figure 5 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS

[0221] Figure 6 shows a matrix plot of synergistic effect of compound RMC-6236 with co-drug Ifebemtinib (top left), compound RMC-6236 with co-drug VS-4718 (top right), compound RMC-6236 with co-drug Defactinib (bottom left) and compound RMC-6236 with co-drug Narmafotinib (bottom right) on KRAS G12V Figure 6 shows a matrix plot of synergistic effect of compound RMC-6236 with co-drug Ifebemtinib (top left), compound RMC-6236 with co-drug VS-4718 (top right), compound RMC-6236 with co-drug Defactinib (bottom left) and compound RMC-6236 with co-drug Narmafotinib (bottom right) on KRAS

[0222] Figure 7 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS G12V Figure 7 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS

[0223] Figure 8 shows a matrix plot of synergistic effect of RMC-6236 with co-drug Ifebemtinib (top left), RMC-6236 with co-drug VS-4718 (top right), RMC-6236 with co-drug Defactinib (bottom left) and RMC-6236 with co-drug Narmafotinib (bottom right) on KRAS G12S Figure 8 shows a matrix plot of synergistic effect of RMC-6236 with co-drug Ifebemtinib (top left), RMC-6236 with co-drug VS-4718 (top right), RMC-6236 with co-drug Defactinib (bottom left) and RMC-6236 with co-drug Narmafotinib (bottom right) on KRAS

[0224] Figure 9 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS G12S Figure 9 shows a matrix plot of synergistic effect of compound PA-1 with co-drug Ifebemtinib (top left), compound PA-1 with co-drug VS-4718 (top right), compound PA-1 with co-drug Defactinib (bottom left) and compound PA-1 with co-drug Narmafotinib (bottom right) on KRAS

[0225] Figure 10 shows a matrix plot of the synergistic effect of compound RMC-6236 in combination with Ifebemtinib (top left), VS-4718 (top right), Defactinib (bottom left) and Narmafotinib (bottom right) on the proliferation inhibition of KRAS wild-type human non-small cell lung cancer H1975 cells.

[0226] Figure 11 shows a matrix plot of the synergistic effect of compound PA-1 in combination with Ifebemtinib (top left), VS-4718 (top right), Defactinib (bottom left) and Narmafotinib (bottom right) on the proliferation inhibition of KRAS wild-type human non-small cell lung cancer H1975 cells.

[0227] Figure 12 shows a matrix plot of the synergistic effect of compound PA-1 in combination with Ifebemtinib (top left), VS-4718 (top right), Defactinib (bottom left) and Narmafotinib (bottom right) on the proliferation inhibition of NRAS G13D human myeloma H929 cells.

[0228] Figure 13 shows a matrix plot of the synergistic effect of RMC-6236 in combination with Ifebemtinib (top left), VS-4718 (top right), Defactinib (bottom left) and Narmafotinib (bottom right) on the proliferation inhibition of HRAS F82L human endometrial carcinoma AN3 CA cells.

[0229] Figure 14 shows a matrix plot of the synergistic effect of compound PA-1 in combination with Ifebemtinib (top left), VS-4718 (top right), Defactinib (bottom left) and Narmafotinib (bottom right) on the proliferation inhibition of HRAS F82L human endometrial carcinoma AN3 CA cells. DETAILED DESCRIPTION

[0230] The compounds disclosed herein can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed herein, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments disclosed herein.

[0231] The present disclosure is described in detail below with reference to embodiments, but this does not imply any adverse limitation thereof. The present disclosure has been described in detail herein, including specific embodiments thereof. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present disclosure without departing from the spirit and scope thereof. All reagents used in this disclosure are commercially available and can be used without further purification.

[0232] Unless otherwise stated, the proportions expressed for mixed solvents are volume-based.

[0233] Unless otherwise stated, % refers to weight percentage (wt%).

[0234] Compounds are processed manually or Software naming conventions are used; commercially available compounds use supplier catalog names.

[0235] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are measured in units of 10⁻⁶. -6 (ppm). The solvents used for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS).

[0236] The eluent or mobile phase may be a mixture of two or more solvents, with the ratio being the volume ratio of each solvent.

[0237] Preparation Example

[0238] Preparation Example 1: Synthesis of intermediate compound Int-1

[0239] Step 1: Synthesis of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropionic acid (compound A2)

[0240] Compound A2 (88 g, 246.82 mmol) was dissolved in dichloromethane (1000 mL) at 0 °C. N,N-dimethylformamide (1.80 g, 24.68 mmol, 1.91 mL) was added to the solution under nitrogen protection. Then oxalyl chloride (62.69 g, 493.65 mmol, 42.13 mL) was added dropwise to the reaction solution. The reaction solution was stirred at 0 °C for 2 hours. The reaction was monitored by LC-MS. The reaction solution was filtered and the filtrate was concentrated under reduced pressure to give compound A3 (80 g, 213.35 mmol, yield: 86.44%) which was used directly in the next step without purification.

[0241] Synthesis of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (Compound A3)

[0242] Compound A3 (80 g, 213.35 mmol) was dissolved in dichloromethane (1.5 L) at 0 °C. Tin tetrachloride solution (1 M, 213.35 mL) and 5-bromo-lH-indole (41.83 g, 213.35 mmol) were added to the solution under nitrogen protection. The reaction solution was reacted at 0 °C for 10 hours. LC-MS showed that the starting material was consumed and the product was detected. The reaction solution was diluted with ethyl acetate (600 mL) and washed with saturated brine (100 mL) four times. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was rotary evaporated to give compound A4 (8 g, 14.97 mmol, yield: 7.01%) which was purified by silica gel chromatography (ethyl acetate / tetrahydrofuran = 5 / 1 to 3 / 1).

[0243] Synthesis of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (Compound A3)

[0244] Compound A3 (80 g, 213.35 mmol) was dissolved in dichloromethane (1.5 L) at 0 °C. Tin tetrachloride solution (1 M, 213.35 mL) and 5-bromo-lH-indole (41.83 g, 213.35 mmol) were added to the solution under nitrogen protection. The reaction solution was reacted at 0 °C for 10 hours. LC-MS showed that the starting material was consumed and the product was detected. The reaction solution was diluted with ethyl acetate (600 mL) and washed with saturated brine (100 mL) four times. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was rotary evaporated to give compound A4 (8 g, 14.97 mmol, yield: 7.01%) which was purified by silica gel chromatography (ethyl acetate / tetrahydrofuran = 5 / 1 to 3 / 1).

[0245] MS (ESI + m / z = 534.0 [M+H] + .

[0246] Step 4: Synthesis of l-(5-bromo-lH-indol-3-yl)-3-((tert-butyldiphenylsilyl)oxy)- 2,2-dimethylpropan-l-ol (Compound A5)

[0247] Compound A4 (8 g, 14.97 mmol) was dissolved in tetrahydrofuran (71.30 mL) at 0 °C, and 2M lithium borohydride tetrahydrofuran solution (2M, 18.71 mL) was added dropwise into the reaction solution under nitrogen protection. Then the reaction solution was heated to 60 °C for 16 hours. LC-MS showed that the starting material was consumed completely and the desired compound was detected. The reaction solution was quenched with methanol (20 mL) and extracted with ethyl acetate (50 mL) for three times. The organic layers were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give Compound A5 (8 g, 14.91 mmol, yield: 99.62%). Without further purification, it was used directly in the next step.

[0248] Step 5: Synthesis of 5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)- lH-indole (Compound A6)

[0249] Compound A5 (8 g, 14.91 mmol), dihydropyridine (4.37 g, 17.25 mmol), p-toluenesulfonic acid monohydrate (2.84 g, 14.91 mmol) were dissolved in dichloromethane (150 mL) and stirred at 0 °C for 2 hours under nitrogen protection. LC-MS showed that the reactants were consumed completely and the desired compound was detected. After the reaction was completed, water (50 mL) was added to quench the reaction, and dichloromethane (50 mL) was used to extract three times. The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. The crude product was purified by silica gel chromatography (petroleum ether / ethyl acetate = 10 / 1) to give Compound A6 (7 g, 13.45 mmol, yield: 90.19%).

[0250] MS (ESI + )m / z = 520.0 [M+H] + .

[0251] Step 6: Synthesis of 5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropyl)- 2-iodo-lH-indole (Compound Int-1)

[0252] Compound A6 (3 g, 5.76 mmol) was dissolved in tetrahydrofuran (10 mL), and I2(1.46 g, 5.76 mmol) and silver trifluoromethanesulfonate (1.78 g, 6.92 mmol) were added. The reaction was stirred at room temperature for 2 hours. LC-MS showed that the reactants were consumed completely, and the desired compound was detected. The reaction was diluted with ethyl acetate (50 mL), washed with saturated Na2S2O3 aqueous solution (50 mL), and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel chromatography (petroleum ether / ethyl acetate = 20 / 1 to 10 / 1) to obtain compound Int-1 (973 mg, 1.51 mmol, yield: 26.12%).

[0253] MS (ESI + )m / z = 646.1 [M+H] + .

[0254] Synthesis of intermediate compound Int-2 in Preparation Example 2

[0255] Synthesis of the first step (4-bromothiazol-2-yl)methanol (compound B2)

[0256] Compound B1 (10 g, 52 mmol) was dissolved in methanol (15 mL), and sodium borohydride (2.95 g, 78.11 mmol) was added, and stirred at 0°C for 0.5 h. Thin layer chromatography showed that compound B1 was completely reacted. The reaction was quenched by adding 10 ml of dilute hydrochloric acid. The reaction mixture was concentrated under reduced pressure to remove the solvent to obtain compound B2 (9 g, 46.38 mmol, yield 89.07%).

[0257] MS (ESI + )m / z = 194.3 [M+H] + .

[0258] Synthesis of the second step 4-bromo-2-(bromoethyl)thiazole (compound B3)

[0259] Carbon tetrabromide (23.07 g, 69.57 mmol), compound B2 (9 g, 46.38 mmol), and triphenylphosphine (18.25 g, 69.57 mmol) were added to dichloromethane (120 mL) at 0°C. After stirring at room temperature for 1 hour, LC-MS showed that the reaction was complete. The mixture was filtered, and the filtrate was concentrated under vacuum, and the crude product was purified by silica gel chromatography (petroleum ether / ethyl acetate = 0-10%) to obtain compound B3 (9.0 g, 35.20 mmol, yield: 75.9%). MS (ESI + )m / z = 255.7 [M+H] + .

[0260] Step 3: Synthesis of 4-bromo-2-[[(2S,5R)-5-isopropyl-3,6-dimethoxy-2,5- dihydropyrazin-2-yl]methyl]thiazole (Compound B5)

[0261] (R)-2,5-dihydro-3,6-dimethoxy-2-isopropylpyrazine (Compound B4, 7.10 g, 38.53 mmol) was added into tetrahydrofuran (100 mL) and n-butyllithium (16.81 mL, 42.03 mmol, 2.5 M) was added slowly at -78 °C. After addition, it was stirred at -78 °C for 0.5 h. Compound B3 (9.0 g, 35.20 mmol) was added into above mixture and stirred at -78 °C for 1 h. LC-MS was used to monitor the completion of reaction. It was quenched with saturated aqueous ammonium chloride solution (30 mL) and extracted with ethyl acetate (100 mL x 2). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column (0-15% petroleum ether / ethyl acetate) to give Compound B5 (10.5 g, 29.14 mmol, yield: 83%).

[0262] MS (ESI + )m / z = 360.2 [M+H] + .

[0263] Step 4: Synthesis of (S)-methyl 2-amino-3-(4-bromothiazol-2-yl)propanoate (Compound B6)

[0264] Compound B5 (10.5 g, 29.14 mmol) was dissolved in acetonitrile (60 mL) and hydrochloric acid (195 mL, 0.3 M) was added. It was stirred at room temperature for 2 h. LC-MS was used to monitor the completion of reaction. The mixture was basified with saturated aqueous sodium bicarbonate solution to pH = 8. It was then extracted with ethyl acetate (100 mL x 6). The organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum to give Compound B6 (6.8 g, 25.65 mmol, yield: 88%).

[0265] MS (ESI + )m / z = 264.9 [M+H] + .

[0266] Step 5: Synthesis of (S)-methyl 3-(4-bromothiazol-2-yl)-2-(tert-butoxycarbonyl)amino)propanoate (Compound B7)

[0267] Triethylamine (8.94 mL, 64.12 mmol) and di-tert-butyl dicarbonate (8.4 g, 38.47 mmol) were added to a solution of compound B6 (6.8 g, 25.65 mmol) in dichloromethane (80 mL) respectively. Stirring at room temperature for 16 hours. LC-MS monitoring reaction complete. Quench with water (75 mL), extract with dichloromethane (75 mL x 2). The organic layer was rotary evaporated and purified by silica gel column (petroleum ether / ethyl acetate = 0-30%) to give compound B7 (6.5 g, yield: 68%).

[0268] MS (ESI + )m / z = 364.9 [M+H] + .

[0269] Synthesis of (S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (compound B8)

[0270] Compound B7 (6.5 g, 17.44 mmol) and lithium hydroxide monohydrate (2.93 g, 69.76 mmol) were added in a mixture solvent of tetrahydrofuran (60 mL), methanol (5 mL) and water (20 mL). Stirring at room temperature for 1 hour. LC-MS monitoring reaction complete. The mixture was acidified to pH = 5 with 1M aqueous hydrochloric acid solution. Extracted with ethyl acetate (100 mL x 2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give compound B8 (6 g, 17.08 mmol, yield: 98%).

[0271] MS (ESI + )m / z = 351.2 [M+H] + .

[0272] Synthesis of (S)-2-((S)-3-(4-bromothiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)-2,3-diazabicyclo[3.1.1]heptane-4-carboxylic acid methyl ester (compound Int-2)

[0273] Compound Int-5 (309 mg, 1.98 mmol) and N,N-diisopropylethylamine (3.45 mL, 19.8 mmol) were added to a solution of compound B8 (695.4 mg, 1.98 mmol) and 2-(7-azobenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (896.3 mg, 2.38 mmol) in N,N-dimethylformamide (7.0 mL) successively, stirring at room temperature for 2 hours. Purified by reverse phase silica gel column (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to give compound Int-2 (600.0 mg, yield: 61.9%).

[0274] MS m / z (ESI): 489.2 [M+H] + .

[0275] Synthesis of intermediate compound Int-3 in Preparation Example 3

[0276] First Step: Synthesis of (S)-1-(3-bromopyridin-2-yl)ethan-1-ol (compound C2)

[0277] A solution of formic acid (6.63 g, 143.98 mmol, 5.43 mL) in triethylamine (72.84 g, 719.88 mmol, 100.41 mL) was cooled to 0 °C under N2protection, then (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanedi-amine (p-cymene) ruthenium chloride (379 mg, 599.90 µmol) was added, the reaction was heated to 40 °C and stirred for 15 min, then cooled to room temperature, compound C1 (12 g, 59.99 mmol) was added, then the reaction was heated to 40 °C and stirred for 2 h. After the reaction was cooled to room temperature, the reaction was concentrated under reduced pressure, and compound C2 (12 g, 59.41 mmol, yield: 99%) was obtained by column chromatography purification. MS (ESI + m / z = 202.1 [M+H] + .

[0278] Second Step: Synthesis of (S)-3-bromo-2-(1-methoxyethyl)pyridine (compound C3)

[0279] A solution of compound C2 (12.00 g, 59.41 mmol) in N,N-dimethylformamide (75 mL) was cooled to 0 °C under N2protection, sodium hydride (2.85 g, 71.27 mmol, 60% purity) was added, the mixture was stirred at 0 °C for 15 min, then iodomethane (16.86 g, 118.78 mmol) was added, and the temperature was allowed to rise to room temperature, and the reaction was stirred for 2 h. The reaction was slowly added to ice water (750 mL), extracted with ethyl acetate (100 mL x 3), the organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated, and the residue was purified by column chromatography to obtain compound C3 (11 g, 50.9 mmol, yield: 86%). MS (ESI + m / z = 216.1 [M+H] + .

[0280] Third Step: Synthesis of 5-bromo-6-[(1S)-1-methoxyethyl]pyridin-3-yl boronic acid (compound C4)

[0281] 4,4'-di-tert-butyl-2,2'-bipyridine (931.61 mg, 3.47 mmol) and 1,5-cyclooctadiene iridium chloride dimer (466.30 mg, 694.20 mmol) were added under a nitrogen atmosphere to a tetrahydrofuran (50 mL) solution of compound C3 (5.0 g, 23.14 mmol) and bis(pinnatrol)boronic acid ester (8.81 g, 34.71 mmol). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 16 hours. LC-MS monitoring showed that the reaction was complete and no starting material remained. The mixture was concentrated under reduced pressure. The resulting mixture was dissolved in ethyl acetate (30 mL), and the pH was adjusted to 10 with a solution of sodium carbonate (40 g) and sodium hydroxide (10 g) (4:1 ratio) in water (600 mL). Extraction was performed with ethyl acetate (100 mL). The aqueous phase was acidified to pH 6 with hydrochloric acid (6M) to give compound C4 (4.5 g, 17.3 mmol, yield: 75%).

[0282] MS(ESI + m / z = 260.0 [M+H] + .

[0283] Step 4: Synthesis of (S)-3-bromo-5-iodo-2-(1-methoxyethyl)pyridine (compound C5)

[0284] Compound C4 (4.5 g, 17.3 mmol) and N-iodosuccinimide (36.70 g, 163.14 mmol) were added to acetonitrile (50 mL) under N2 protection. The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 16 hours. The reaction was monitored by LC-MS until complete. The resulting mixture was dissolved in dichloromethane (80 mL), washed with saturated sodium thiosulfate aqueous solution (80 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 0-15%) to give compound C5 (4.3 g, 12.6 mmol, yield: 73%).

[0285] MS(ESI + m / z = 341.8[M+H] + .

[0286] Step 5: Synthesis of (S)-4-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (compound C7)

[0287] A mixture of compound C5 (4.3 g, 12.6 mmol), compound C6 (2.77 g, 12.57 mmol), (R)-(+)-2,2-bis(diphenylphosphino)-1,1-naphthylhydride (156.59 mg, 251.48 µmol), palladium acetate (141.15 mg, 628.71 µmol), Cs2CO3 (10.24 g, 31.44 mmol), toluene (50 mL) was placed in a sealed tube under N2protection. The resulting solution was stirred at 100 °C under nitrogen atmosphere for 16 hours. The reaction was monitored by LC-MS until completion. After the reaction was completed, the reaction mixture was cooled to room temperature. The mixture was extracted with ethyl acetate (80 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column (petroleum ether / ethyl acetate: 0-45%) to give compound C7 (3.6 g, 8.29 mmol, yield: 65.92%).

[0288] MS (ESI + )m / z = 434.2 [M+H] + .

[0289] Synthesis of (S)-4-(6-(1-methoxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (compound Int-3)

[0290] A mixture of compound C7 (9.6 g, 22.10 mmol), B2Pin2 (28.06 g, 110.52 mmol), potassium acetate (6.51 g, 66.31 mmol) was dissolved in dioxane (100 mL) and Pd(dppf)Cl2 (1.62 g, 2.21 mmol) was added to the reaction solution under nitrogen protection and replaced with nitrogen for 5 times, stirred at 100 °C for 16 hours. The reaction was monitored until completion. After filtration, the reaction mixture was concentrated under reduced pressure, ethyl acetate (100 mL) and 6N HCl (100 mL) were added and stirred for 16 hours, filtered, concentrated under reduced pressure, and purified by reverse phase silica gel column (water / acetonitrile = 1 / 0~1 / 1) to give compound Int-3 (4.6 g, 9.52 mmol, yield: 43.06%). MS (ESI + )m / z = 482.2 [M+H] + .

[0291] Synthesis of intermediate compound Int-4 in Preparation 4

[0292] Step 1: Synthesis of (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2- dimethylpropyl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)benzyl piperazine-1- carboxylate (Compound D1)

[0293] A solution of compound Int-1 (4 g, 6.19 mmol), compound Int-3 (4.47 g, 9.28 mmol), potassium carbonate (2.57 g, 18.56 mmol) in ethylene glycol dimethyl ether (40 mL) and water (8 mL) was stirred at 100 °C for 16 h under nitrogen atmosphere. The reaction was monitored by LC-MS. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 1 / 0 ~ 1 / 5) to give compound D1 (2.3 g, 2.63 mmol, 42.49% yield).

[0294] MS (ESI + )m / z = 873.3 [M+H] + .

[0295] Step 2: Synthesis of (S)-4-(5-(5-bromo-3-(3-((tert-butyldiphenylsilyl)oxy)-2,2- dimethylpropyl)-1-ethyl-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)benzyl piperazine-1- carboxylate (Compound D2)

[0296] A solution of compound D1 (2.3 g, 2.63 mmol) in DMF (30 mL) was stirred at 25 °C for 16 h after the addition of cesium carbonate (2.57 g, 7.89 mmol) and iodoethane (820.88 mg, 5.26 mmol) portionwise. The reaction was monitored by LC-MS. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound D2 (2.0 g, crude). It was used directly in the next step.

[0297] MS (ESI + )m / z = 901.3 [M+H] + .

[0298] Step 3: Synthesis of (S)-4-(5-(5-bromo-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H- indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)benzyl piperazine-1-carboxylate (Compound D3)

[0299] Compound D2 (2.0 g, crude) was dissolved in tetrahydrofuran (30 mL), TBAF (1 M, 22.17 mL) was added, and the mixture was stirred at 25 °C for 16 h. LC-MS was used to monitor the completion of the reaction. The mixture was concentrated under reduced pressure and purified by a normal silica gel column (petroleum ether / ethyl acetate = 1 / 0 ~ 0 / 1) to give the less polar isomer, which was Compound D3 (0.5 g, 753.42 μmol, yield: 33.4%).

[0300] MS (ESI + )m / z = 663.3 [M+H] + .

[0301] Synthesis of (S)-4-(5-(1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-2-yl)-6-(1-methoxyethyl)pyridin-3-yl)piperazine-1-carboxylic acid benzyl ester (Compound D4)

[0302] Compound D3 (0.5 g, 753.42 μmol), bis(pinacolato)diboron (573.96 mg, 2.26 mmol), potassium acetate (184.86 mg, 1.88 mmol) were dissolved in toluene (10 mL), Pd(dppf)Cl2 (55.13 mg, 75.34 μmol) was added to the reaction solution under nitrogen protection and replaced with nitrogen for 5 times, and the mixture was stirred at 100 °C for 16 h. LC-MS was used to monitor the completion of the reaction. The mixture was filtered and concentrated under reduced pressure, diluted with water (50 mL), extracted with ethyl acetate (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by a normal silica gel column (petroleum ether / ethyl acetate = 1 / 0 ~ 1 / 3) to give Compound D4 (0.4 g, 562.82 μmol, yield: 74.7%).

[0303] MS (ESI + )m / z = 711.4 [M+H] + .

[0304] Synthesis of (S)-2-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1-methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoyl)-2,3-diazabicyclo[3.1.1]heptane-4-carboxylic acid methyl ester (Compound D5)

[0305] Compound D4 (117.6 mg, 165.5 μmol), compound Int-2 (89 mg, 182.05 μmol) and potassium phosphate (232.96 mg, 1.10 mmol) were dissolved in a mixed solution of dioxane (1 mL), toluene (3 mL) and water (1 mL), Pd(dtbpf)Cl2 (23.84 mg, 36.58 μmol) was added into the reaction solution under nitrogen protection condition and replaced by nitrogen for 5 times, stirred at 100 °C for 16 hours, LC-MS was used to monitor the completion of the reaction. Diluted with water (50 mL), extracted with ethyl acetate (10 mL x 3), washed with anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, purified by normal phase silica gel column (petroleum ether / ethyl acetate = 1 / 0 ~ 1 / 3) to obtain compound D5 (142.2 mg, 143.16 μmol, yield: 86.5%).

[0306] MS (ESI + )m / z = 993.4 [M+H] + .

[0307] Synthesis of (S)-2-((S)-3-(4-(2-(5-(4-((benzyloxy)carbonyl)piperazin-1-yl)-2-((S)-1- methoxyethyl)pyridin-3-yl)-1-ethyl-3-(3-hydroxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)- 2-((tert-butoxycarbonyl)amino)propanoyl)-2,3-diazabicyclo[3.1.1]octane-4-carboxylic acid (Compound D6)

[0308] Compound D5 (142.2 mg, 143.16 μmol) was dissolved in a mixed solution of tetrahydrofuran (5 mL) and water (5 mL), lithium hydroxide (32.95 mg, 1.37 mmol) was added into the reaction solution, stirred at room temperature for 16 hours, LC-MS was used to monitor the completion of the reaction. Diluted with ethyl acetate (30 mL) and water (30 mL), the aqueous phase was adjusted to about pH 6 with 1M aqueous hydrochloric acid solution, extracted with ethyl acetate (30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to obtain yellow solid compound D6 (120 mg, 122.83 μmol, yield: 85.8%).

[0309] MS (ESI + )m / z = 979.5 [M+H] + .

[0310] Synthesis of compound D7

[0311] Compound D6 (120 mg, 122.83 μmol), DIEA (1.12 g, 8.69 mmol, 1.51 mL), EDCI (1.25 g, 6.51 mmol) and HOBt (293.39 mg, 2.17 mmol) were dissolved in acetonitrile (5 mL), stirred at room temperature for 16 hours, and the reaction was monitored to completion by LC-MS. Diluted with water (20 mL), extracted with ethyl acetate (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by normal phase silica gel column (petroleum ether / ethyl acetate = 1 / 0 ~ 1 / 4) to give compound D7 (35.4 mg, 36.85 μmol, yield: 30.4%).

[0312] MS (ESI + )m / z = 961.1 [M+H] + .

[0313] Synthesis of compound D8 in the eighth step

[0314] Compound D7 (35.4 mg, 36.85 μmol) was dissolved in methanol (10 mL), Pd(OH)2 / C (64.11 mg, 273.92 μmol, 60% purity) was added to the reaction solution under nitrogen protection, and then the nitrogen was replaced with hydrogen for 5 times, stirred at room temperature for 16 hours under one atmosphere, and the reaction was monitored to completion by LC-MS. Filtration and concentration under reduced pressure gave compound D8 (28 mg, 33.9 μmol, yield: 92%).

[0315] MS (ESI + )m / z = 827.2 [M+H] + .

[0316] Synthesis of compound D9 in the ninth step

[0317] Compound D8 (28 mg, 33.9 μmol) and acetic acid (22.10 mg, 368.08 μmol) were dissolved in methanol (5 mL), stirred at room temperature for 1 hour, then paraformaldehyde (36.85 mg, 1.23 mmol) and NaBH3CN (23.13 mg, 368.08 μmol) were added to the reaction solution, stirred at room temperature for 16 hours, and the reaction was monitored to completion by LC-MS. Filtration and concentration under reduced pressure gave compound D9 (24 mg, 28.53 μmol, yield: 84.16%).

[0318] MS (ESI + )m / z = 841.2 [M+H] + .

[0319] Synthesis of compound Int-4 in the tenth step

[0320] 2Compound D9 (14 mg, 16.7 μmol) was dissolved in methanol (10 mL), 4N hydrogen chloride in dioxane (5 mL) was added to the reaction under nitrogen protection, stirred at room temperature for 5 hours, LC-MS monitoring reaction completion, rotary evaporation to concentrate to obtain compound Int-4 (10 mg, 13.54 μmol, yield: 81.1%). MS (ESI + m / z = 741.1 [M+H] + .

[0321] Synthesis of intermediate compound Int-5 in Preparation Example 5

[0322] Synthesis of first step 3-(2-diazoacetyl)cyclobutan-l-one (compound E2)

[0323] Dichlorosulfoxide (89.12 mL, 1.23 mol) was added dropwise to a solution of compound El (70.0 g, 613.5 mmol) in ethyl acetate (700.0 mL) under ice bath. The mixture was heated to 60 °C and stirred for 4 hours. After the reaction was completed, the reaction was concentrated to dryness and azeotroped with toluene. The obtained crude product was dissolved in a mixture of tetrahydrofuran (250.0 mL) and acetonitrile (250.0 mL). 2.0 M trimethylsilyl diazomethane in hexane (460.1 mL, 920.2 mmol) was added dropwise to the solution of the crude product at 0 °C, slowly warmed to room temperature and stirred for 12 hours. After the reaction was completed, the reaction was cooled to 0 °C and quenched by the addition of acetic acid (50.0 mL) and water (200.0 mL), then concentrated to obtain a residue, which was diluted with saturated aqueous sodium bicarbonate solution (200.0 mL). The obtained mixture was extracted with ethyl acetate (300 mL) three times. The combined organic layer was washed with saturated aqueous sodium chloride solution (300 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 1) to obtain compound E2 (45.0 g, 325.77 mmol, yield: 53.1%).

[0324] MS m / z (ESI): 139.0 [M+H] + .

[0325] Preparation of second step 2-(3-oxocyclobutyl)acetic acid (compound E3)

[0326] Silver nitrate (59.0 g, 347.5 mmol) was added portionwise to a mixture of compound E2 (40.0 g, 289.6 mmol) in water (360.0 mL) and tetrahydrofuran (720.0 mL) and stirred at room temperature for 12 h. After completion of the reaction, the reaction mixture was concentrated to get a residue, to which water (1000.0 mL) was added and the pH was adjusted to 1-2 with dilute hydrochloric acid (1.0 M) and the resulting mixture was extracted with ethyl acetate (300 mL) five times. The combined organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to get compound E3 (36.0 g, 280.91 mmol, yield: 97.0%) which was used as such for the next step without further purification.

[0327] MS m / z (ESI): 127.0 [M-H] - .

[0328] Preparation of (S)-4-benzyl-3-(2-(3-oxocyclobutyl)acetyl)oxazolidin-2-one (compound E4)

[0329] Compound E3 (36 g, 280.91 mmol), (S)-4-benzyl oxazolidin-2-one (49.8 g, 281.0 mmol), 4-dimethylaminopyridine (3.8 g, 31.2 mmol) and triethylamine (130.6 mL, 936.6 mmol) were added to dichloromethane (800.0 mL) one by one followed by the addition of 2-chloro-l-methylpyridinium iodide (87.7 g, 343.4 mmol) portionwise. It was stirred at room temperature for 1 h. After completion of the reaction, the reaction was quenched with water and the organic layer was washed with water (1000.0 mL) twice, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to get a residue which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to get compound E4 (49.43 g, 171.64 mmol, yield: 61.1%).

[0330] MS m / z (ESI): 288.1 [M+H] + .

[0331] Preparation of (S)-4-benzyl-3-(2-(3-oxocyclobutyl)acetyl)oxazolidin-2-one (compound E4)

[0332] Compound E4 (49.43 g, 171.64 mmol) and acetic acid (22.9 g, 381.4 mmol) were added to tetrahydrofuran (550.0 mL) sequentially, and the temperature was lowered to 0 °C. Sodium borohydride (5.77 g, 152.6 mmol) was added in portions, and after the addition was completed, the mixture was stirred for 2 hours. After the reaction was completed, saturated ammonium chloride aqueous solution (150.0 mL) was slowly added dropwise to quench the reaction, and the residue was concentrated under reduced pressure. The residue was extracted with ethyl acetate (300.0 mL) three times, and the organic phase was washed with saturated sodium bicarbonate aqueous solution, the pH was adjusted to 8, the organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain compound E5 (48.68 g, 167.86 mmol, yield: 97.8%), which was used in the next step without further purification.

[0333] MS m / z (ESI): 290.2 [M+H] + .

[0334] Preparation of (S)-3-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)cyclobutyl-4- methylbenzenesulfonate (Compound E6)

[0335] Compound E5 (48.68 g, 167.86 mmol), 4-dimethylaminopyridine (18.2 g, 149.3 mmol), and N,N-diisopropylethylamine (48.8 mL, 280.0 mmol) were added to anhydrous dichloromethane (500.0 mL), and the temperature was lowered to 0 °C. p-Toluenesulfonyl chloride (39.1 g, 205.3 mmol) was added in portions, and after the addition was completed, the reaction mixture was slowly warmed to room temperature and stirred overnight. After the reaction was completed, the mixture was washed with water (500.0 mL) and extracted with dichloromethane. The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound E6 (49.8 g, 112.14 mmol, yield: 66.8%).

[0336] MS m / z (ESI): 444.1 [M+H] + .

[0337] Preparation of (S)-4-benzyl-3-(2-(3-bromocyclobutyl)acetyl)oxazolidin-2-one (Compound E7)

[0338] Compound E6 (49.8 g, 112.14 mmol) and lithium bromide (19.0 g, 219.2 mmol) were added to N-methylpyrrolidone (500.0 mL), and the reaction liquid was heated to 90°C and stirred for 12 hours. After the reaction was completed, saturated aqueous sodium chloride solution (1.0 L) was added for dilution, and ethyl acetate (300.0 mL) was extracted three times, and the organic phase was washed once more with saturated aqueous sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound E7 (34.74 g, 98.68 mmol, yield: 88.0%). MS m / z (ESI): 352.2 [M+H] + .

[0339] Seventh step: Preparation of (S)-2,3-bis(tert-butoxycarbonyl)-2,3-diazabicyclo[3.1.1]heptane-4-carboxylic acid (compound E8)

[0340] Compound E7 (10.0 g, 28.4 mmol) was dissolved in tetrahydrofuran (100.0 mL) under an argon atmosphere, and cooled to -78°C, and then a solution of diisopropylamino lithium in tetrahydrofuran n-heptane (18.5 mL, 2.0 M) was slowly added dropwise, and stirred for 0.5 hours. Then, di-tert-butyl azodicarboxylate (7.84 g, 34.0 mmol) in anhydrous dichloromethane (20.0 mL) was added to the above solution, and stirring was continued for 0.5 hours. Next, N,N-dimethylpropenylurea (109.2 g, 851.7 mmol) was slowly added to the above reaction liquid, and slowly warmed to room temperature and stirring was continued for 13 hours. After the reaction was completed, water (100.0 mL) was added to quench the reaction, and lithium hydroxide monohydrate (3.58 g, 85.1 mmol) was added, and stirred at room temperature for 1 hour. After the reaction was completed, the reaction liquid was concentrated, and then saturated aqueous sodium chloride solution (200.0 mL) was added for dilution, and extracted with ethyl acetate (200.0 mL) three times, and the organic phase was discarded, and the aqueous phase was adjusted to pH 5 with dilute hydrochloric acid (1.0 M), and extracted with ethyl acetate (200.0 mL) three times, and the organic phase was washed once with saturated aqueous sodium chloride solution, and the organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound E8 (1.0 g, 2.91 mmol, yield: 10.2%).

[0341] MS m / z (ESI): 343.1 [M+H] + .

[0342] Step 8 Preparation of 2,3-di-tert-butyl-4-methyl (S)-2,3-diazabicyclo[3.1.1]heptane-2,3,4- tricarboxylate (Compound E9)

[0343] To a solution of compound E8 (1.0 g, 2.91 mmol) in methanol (10.0 mL) was added slowly dropwise a solution of trimethylsilyldiazomethane in n-hexane (7.3 mL, 2.0 M) at room temperature, and the mixture was stirred at room temperature for 30 minutes. After the reaction was completed, a few drops of acetic acid were added to quench the reaction. The reaction solution was concentrated to obtain the title compound E9 (1.0 g, 2.8 mmol, yield: 96.2%).

[0344] MS m / z (ESI): 357.2 [M+H] + .

[0345] Step 9 Preparation of (S)-methyl 2,3-diazabicyclo[3.1.1]heptane-4-carboxylate (Compound Int-5)

[0346] To a solution of compound E9 (706.0 mg, 1.98 mmol) in dichloromethane (6.0 mL) was added slowly dropwise trifluoroacetic acid (2.0 mL) at room temperature, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was concentrated to obtain the title compound Int-5 (312 mg, 1.98 mmol, yield: 100.0%).

[0347] MS m / z (ESI): 157.0 [M+H] + .

[0348] Synthesis of intermediate compound Int-6 in Preparation Example 6

[0349] Step 1 Synthesis of (S)-4-(3-(5-bromo-6-(1-methoxyethyl)pyridin-3-yl)prop-2-yn-1-yl)morpholine (Compound Int-6)

[0350] To a solution of compound C5 (3.4 g, 10.0 mmol) dissolved in tetrahydrofuran (20 mL) were sequentially added cuprous iodide (190 mg, 1.0 mmol), triethylamine (2.02 g, 20.0 mmol), dichlorobis(triphenylphosphine)palladium (702 mg, 1.0 mmol), 4-propyn-1-morpholine (1.88 g, 15.0 mmol), and the reaction mixture was reacted at room temperature for 5 hours under argon protection. After the reaction was completed, the organic solvent was removed by distillation under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:3) to obtain compound Int-6 (2.5 g, 7.3 mmol, yield: 73%).

[0351] MS m / z (ESI): 339.1 [M+H] + .

[0352] Synthesis of compound 1 of Example 1

[0353] To a solution of compound Int-4 (10 mg, 13.54 μmol), N,N- diisopropylethylamine (23.5 μL, 135.0 μmol) in N,N-dimethylformamide (1.0 mL) was added 2-ethylbutyric acid 1A (3.1 mg, 27.0 μmol) and (2- hydroxyimino-cyanoacetic acid ethyl ester)-N,N-dimethyl-morpholino urea hexafluorophosphate (11.5 mg, 27.0 μmol) successively, stirred at room temperature for 1 hour. Ethyl acetate (5 mL) and water (5 mL) were added to extract, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase column (column: Boston Prime C18; 150*30mm*5μm; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration was 0.05%, NH4HCO3 concentration was 2mM; mobile phase B: MeCN; MeCN ratio 40%-60%) to give compound 1 (8.5 mg, 10.16 μmol, yield: 75%). MS (ESI m / z = 839.5 [M+H] + ) + .

[0354] 1H NMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 2.8 Hz, 1H), 8.42 (s, 1H), 8.34 (d, J = 8.6 Hz, 1H), 7.83 (s, 1H), 7.74 (d, J = 8.6 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.23 (s, 1H), 5.92 (d, J = 11.1 Hz, 1H), 5.36 - 5.32 (m, 2H), 4.74 (d, J = 11.0 Hz, 1H), 4.53 - 4.49 (m, 1H), 4.33 - 4.28 (m, 1H), 4.21 - 4.12 (m, 2H), 3.58 - 3.53 (m, 2H), 3.30 - 3.27 (m, 3H), 3.26 - 3.24 (m, 1H), 3.22 (s, 3H), 2.95 (d, J = 14.3 Hz, 1H), 2.69 - 2.65 (m, 2H), 2.37 - 2.32 (m, 1H), 2.25 (s, 3H), 2.18 - 2.13 (m, 2H), 2.06 - 2.01 (m, 3H), 1.64 - 1.58 (m, 1H), 1.47 - 1.40 (m, 6H), 1.34 (d, J = 6.0 Hz, 3H), 0.96 - 0.91 (m, 9H), 0.90 - 0.84 (m, 7H).

[0355] Synthesis of compound 2 of example 2

[0356] Synthesis of compound 2-2, ethyl 2-((2S,5S)-2,5-dimethylpyrrolidin-1-yl)-2- oxoacetate

[0357] Compound 2-1 (1 g, 10.1 mmol), triethylamine (1.42 g, 14.06 mmol, 1.96 mL) were dissolved in dichloromethane (20 mL) and stirred, compound 2A (1.5 g, 11 mmol) was added dropwise to the reaction liquid at 0 °C, then stirred at 25 °C for 2 hours, LC-MS was used to monitor the completion of the reaction. Ethyl acetate (100 mL) and water (100 mL) were added to extract, the organic phase was washed with saturated brine (100 mL), dried by adding anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-2 (0.86 g, 4.3 mmol, yield: 42.57%).

[0358] MS (ESI + )m / z = 200.2 [M+H] + .

[0359] Synthesis of compound 2-3, 2-((2S,5S)-2,5-dimethylpyrrolidin-1-yl)-2-oxoacetic acid

[0360] Compound 2-2 (0.86 g, 4.3 mmol) was dissolved in tetrahydrofuran (5 mL), LiOH.H2O (629.51 mg, 15 mmol) was dissolved in water (5 mL) and added to the reaction solution, which was stirred at room temperature for 1 hour. The reaction was monitored by LC-MS. Ethyl acetate (30 mL) and water (30 mL) were added to extract the reaction solution. The aqueous phase was adjusted to a pH of about 2 with 6M hydrochloric acid, and ethyl acetate (30 mL) was added to extract the organic phase. The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 2-3 (460 mg, 2.68 mmol, yield: 62.3%).

[0361] MS (ESI + )m / z = 172.0 [M+H] + .

[0362] Synthesis of compound 2 of the third step

[0363] Compound 2-3 (5.68 mg, 32.92 μmol), compound Int-4 (10 mg, 13.54 μmol) were dissolved in DMF (1 mL), and DIEA (21.28 mg, 164.62 μmol, 28.67 μL), COMU (28.20 mg, 65.85 μmol) were added portionwise. The reaction was stirred at 25°C for 2 hours. The reaction was monitored to be complete. Ethyl acetate (5 mL) and water (5 mL) were added to extract the reaction solution. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by a reverse phase column (Boston Prime C18; 150*30 mm*5 μm; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration of 0.05%, NH4HCO3 concentration of 2 mM; mobile phase B: MeCN; MeCN ratio 40%-60%) to obtain compound 2 (4.2 mg, 4.69 μmol, yield: 34.63%).

[0364] MS (ESI + )m / z = 894.3 [M+H] + .

[0365] 1H NMR (400 MHz, DMSO-d6) δ 9.07 (d, J = 8.3 Hz, 1H), 8.44 (s, 1H), 8.41 (s, 1H), 7.84 (s, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.56 (d, J = 8.5 Hz, 1H), 7.22 (s, 1H), 6.06 (d, J = 11.6 Hz, 1H), 4.72 (d, J = 11.1 Hz, 1H), 4.54 - 4.50 (m, 2H), 4.38 - 4.23 (m, 1H), 4.21 - 4.08 (m, 3H), 3.60 - 3.50 (m, 2H), 3.48 - 3.42 (m, 2H), 3.21 (s, 3H), 2.92 (d, J = 14.3 Hz, 1H), 2.69 - 2.64 (m, 1H), 2.37 - 2.31 (m, 1H), 2.21 (s, 3H), 2.03 - 1.95 (m, 6H), 1.65 - 1.61 (m, 1H), 1.56 - 1.51 (m, 1H), 1.48 - 1.43 (m, 6H), 1.33 (d, J = 6.0 Hz, 3H), 1.16 (d, J = 6.4 Hz, 3H), 1.08 (d, J = 6.5 Hz, 3H), 0.93 - 0.87 (m, 6H), 0.86 - 0.81 (m, 6H).

[0366] Synthesis of compound 3 of Example 3

[0367] First Step: Synthesis of 3-(5-bromo-lH-indol-3-yl)-2,2-dimethyl-l-propanol (Compound A6-1)

[0368] Compound A6 (10.4 g, 20.0 mmol) was dissolved in tetrahydrofuran (20 mL) at room temperature, and a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.0 M, 50.0 mL) was added. The mixed solution was stirred at 60 °C for 16 hours. After the reaction was completed, the reaction solution was added dropwise to water to quench the reaction, and extracted with dichloromethane, and the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:3) to obtain the title compound A6-1 (4.67 g, 16.6 mmol, yield: 83.0%).

[0369] MS m / z (ESI): 282.1 [M+H] + .

[0370] Second Step: Synthesis of 3-(5-bromo-lH-indol-3-yl)-2,2-dimethylpropyl acetate (Compound 3-1)

[0371] To a solution of compound A6-1 (3.7 g, 13.11 mmol), 4-dimethylaminopyridine (80.1 mg, 655.6 µmol) and triethylamine (3.98 g, 39.34 mmol) in dichloromethane (40.0 mL) was added acetic anhydride (1.28 mL, 13.11 mmol) dropwise under ice-bath. The mixture was slowly warmed to room temperature and stirred for 6 hours. After the reaction was completed, the reaction solution was added dropwise to ice water to quench the reaction, and extracted with dichloromethane, and the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain the title compound 3-1 (4.0 g, 12.34 mmol, yield: 94.0%).

[0372] MS m / z (ESI): 324.1 [M+H] + .

[0373] Third step: Preparation of propyl 2,2-dimethyl-3-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)acetate (compound 3-2)

[0374] To a solution of compound 3-1 (4.6 g, 14.19 mmol), potassium acetate (3.48 g, 35.47 mmol) and bis(pinacolato)diboron (9.0 g, 35.47 mmol) in 1,4-dioxane (46.0 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (1.04 g, 1.42 mmol), and purged with argon three times. The resulting mixture was heated to 90℃ under argon protection and stirred for 3 hours. After the reaction was completed, the reaction solution was filtered with celite, and washed with ethyl acetate, and the filtrate was concentrated under reduced pressure to obtain a residue, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain the title compound 3-2 (4.5 g, 12.12 mmol, yield: 85.4%).

[0375] MS m / z (ESI): 372.1 [M-H] + .

[0376] Fourth step: Preparation of methyl (S)-3-(4-(3-(3-acetyloxy-2,2-dimethylpropyl)-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propanoate (compound 3-3)

[0377] Compound 3-2 (2.6 g, 7.0 mmol), compound B7 (2.81 g, 7.7 mmol) and potassium phosphate (3.71 g, 17.5 mmol) were dissolved in a mixed solution of dioxane (30.0 mL) and water (3.0 mL), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) (452.2 mg, 618 μmol) was added to the reaction solution under nitrogen protection and replaced with argon 5 times, stirred at 90°C for 12 hours, and the reaction was monitored by LC-MS. The reaction was diluted with water (50 mL) and extracted with ethyl acetate (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to obtain compound 3-3 (3.0 g, 5.66 mmol, yield: 80.9%).

[0378] MS m / z (ESI): 530.1 [M+H] + .

[0379] Fifth step: Preparation of (S)-3-(4-(3-(3-acetyloxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)thiazol-2-yl)-2-((tert-butoxycarbonyl)amino)propionic acid methyl ester (compound 3-4)

[0380] Compound 3-3 (3.7 g, 6.98 mmol) and N-iodosuccinimide (1.57 g, 6.99 mmol) were added to N,N-dimethylformamide (40.0 mL) and warmed to 50°C and stirred for 2 hours. After the reaction was completed, the reaction solution was poured into water (400.0 mL) and extracted with ethyl acetate (50.0 mL) three times, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the crude product was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to obtain the title compound 3-4 (2.6 g, 3.97 mmol, yield: 56.77%).

[0381] MS m / z (ESI): 656.5 [M-H] + .

[0382] Sixth step: Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(3-(3-hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)thiazol-2-yl)propionic acid (compound 3-5)

[0383] Compound 3-4 (2.6 g, 3.97 mmol) was dissolved in a mixture solution of tetrahydrofuran (30 mL) and water (5 mL), lithium hydroxide (474.9 mg, 19.8 mmol) was added to the reaction solution, stirred at 25 °C for 16 hours, LC-MS was used to monitor the completion of the reaction, the organic solvent was removed by reduced pressure distillation, diluted with ethyl acetate and water, the aqueous phase was adjusted to about pH 6 with 1M aqueous HC1 solution, ethyl acetate (30.0 mL) was added to extract, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 3-5 (2.3 g, 3.84 mmol, yield: 96.7%).

[0384] MS m / z (ESI): 600.0 [M+H] + .

[0385] Seventh step: Preparation of (S)-methyl 2-((S)-2-(tert-butoxycarbonyl)amino)-3-(4-(3-(3- hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)thiazol-2-yl)propanoate (Compound 3-6)

[0386] Compound 3-5 (393.0 mg, 655.2 μmol), compound Int-5 (265.0 mg, 1.7 mmol), N,N- diisopropylethylamine (1.69 g, 13.11 mmol) and 2-(7-azabenzotriazol-1-yl)-1,1,3,3- tetramethyluronium hexafluorophosphate (370.8 mg, 975.3 μmol) were added to a solution of N,N- dimethylformamide (8.0 mL), stirred at room temperature for 2 hours. After the reaction was completed, the crude product was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to obtain the title compound 3-6 (290.0 mg, 393.1 μmol, yield: 60.0%).

[0387] MS m / z (ESI): 738.1 [M+H] + .

[0388] Eighth step: Preparation of (S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-(4-(3-(3- hydroxy-2,2-dimethylpropyl)-2-iodo-1H-indol-5-yl)thiazol-2-yl)propanoyl)-2,3- diazabicyclo[3.1.1]octane-4-carboxylic acid (Compound 3-7)

[0389] Compound 3-6 (290 mg, 393.1 μmol) was dissolved in a mixture of tetrahydrofuran (2.0 mL) and water (2.0 mL), and lithium hydroxide (94.1 mg, 3.93 mmol) was added to the reaction solution. The mixture was stirred at 25 °C for 2 hours. The reaction was monitored by LC-MS. When the reaction was completed, the reaction solution was diluted with ethyl acetate and water. The aqueous phase was adjusted to pH 6 with 1M aqueous HC1 solution. Ethyl acetate was added to the reaction solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 3-7 (200.0 mg, 276.4 μmol, yield: 70.3%).

[0390] MS m / z (ESI): 724.1 [M+H] + .

[0391] Ninth step: Preparation of compound 3-8

[0392] Compound 3-7 (140.0 mg, 193.4 μmol), N-methylimidazole (794.2 mg, 9.67 mmol), and N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (298.5 mg, 1.06 mmol) were added to a mixture of N,N-dimethylformamide (1.2 mL) and acetonitrile (12.0 mL), and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the crude product was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to obtain the title compound 3-8 (44.0 mg, 62.3 μmol, yield: 32.2%).

[0393] MS m / z (ESI): 706.2 [M+H] + .

[0394] Tenth step: Preparation of compound 3-9

[0395] Compound 3-8 (98.0 mg, 138.0 μmol), potassium acetate (19.47 mg, 198.4 μmol), 2-dicyclohexylphosphino-2',6'-dimethoxy-biphenyl (11.6 mg, 28.3 μmol) and tris(dibenzylideneacetone)dipalladium (10.3 mg, 11.3 μmol) were added to 1,4-dioxane (4.0 mL) and purged with argon three times. The mixture was cooled to 0 °C in an ice bath and a solution of pinacolborane (58.0 mg, 453.5 μmol) in 1,4-dioxane (4.0 mL) was added dropwise to the mixture under argon atmosphere. The reaction was then heated to 50 °C and stirred for 3 h. After completion of the reaction, the reaction mixture was filtered over celite and the filtrate was washed with saturated aqueous sodium chloride solution and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude obtained was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to give the title compound 3-9 (25.0 mg, 35.4 μmol, 25.6% yield).

[0396] MS m / z (ESI): 706.1 [M+H] + .

[0397] Eleventh step: Preparation of compound 3-10

[0398] Compound 3-9 (25.0 mg, 35.4 μmol), compound Int-6 (18.0 mg, 53.1 μmol) and potassium carbonate (14.7 mg, 106.2 μmol) were added to a mixture of 1,4-dioxane (3.6 mL) and water (0.9 mL) and purged with argon. [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(ll) (5.2 mg, 7.1 μmol) was added under argon atmosphere and the resulting mixture was heated to 70 °C for 16 h under argon. After completion of the reaction, the reaction mixture was filtered over celite and the filtrate was washed with saturated aqueous sodium chloride solution and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude obtained was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to give the title compound 3-10 (21.0 mg, 25.0 μmol, 70.7% yield).

[0399] MS m / z (ESI): 838.2 [M+H] + .

[0400] Twelfth step: Preparation of compound 3-11

[0401] Compound 3-10 (25.0 mg, 29.8 pmol) and cesium carbonate (24.5 mg, 75.2 pmol) were added to N,N-dimethylformamide (2.0 mL) and argon was replaced. To the reaction solution, iodoe thane (5.8 mg, 37.5 pmol) was added dropwise under argon atmosphere, and the resulting mixture was stirred at room temperature for 3 hours. After the reaction was completed, the organic phase was dried and concentrated to obtain the crude product, which was purified by reverse phase column chromatography (water: acetonitrile, gradient: 95 / 5 to 5 / 95) to obtain the title compound 3-11 (16.0 mg, 18.4 pmol, yield: 61.7%).

[0402] MS m / z (ESI): 766.2 [M+H] + .

[0403] Thirteenth step: Preparation of compound 3-12

[0404] To a solution of compound 3-11 (16.0 mg, 18.4 pmol) in dichloromethane (2.0 mL) was slowly added trifluoroacetic acid (0.7 mL) dropwise at room temperature, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated to dryness and washed with an aqueous sodium bicarbonate solution, extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the title compound 3-12 (14.0 mg, 18.2 pmol, yield: 98.9%).

[0405] MS m / z (ESI): 766.2 [M+H] + .

[0406] Fourteenth step: Preparation of compounds 3-P1 and 3-P2

[0407] Compound 3-12 (14.0 mg, 18.2 pmol) and N,N-diisopropylethylamine (23.6 pL, 135.0 pmol) were sequentially added to a solution of 2-ethylbutanoic acid (6.3 mg, 54.8 pmol) and 2-(7-azobenzo-triazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (8.2 mg, 21.9 pmol) in N,N-dimethylformamide (1.0 mL), and stirred at room temperature for 1 hour. After the reaction was completed, compounds 3-P1 (3.5 mg, 4.0 pmol, yield: 22.1%, retention time: 8.5 min) and 3-P2 (3.0 mg, 3.4 pmol, yield: 19.0%, retention time: 9.07 min) were obtained after purification by reverse phase column (Boston Prime C18; 150*30 mm*5 pm; mobile phase A: H2O-(NH3H2O), NH3H2O concentration 0.05%; mobile phase B: MeCN; MeCN ratio 50%-75%).

[0408] Compound 3-P1 MS (ESI + m / z = 864.3 [M+H] + .

[0409] 1 H NMR (400 MHz, CDC13) δ 8.84 (d, J = 2.0 Hz, 1H), 8.41 (d, J = 1.5 Hz, 1H), 7.69 (d, J = 2.1 Hz, 1H), 7.61 (dd, J = 8.6, 1.6 Hz, 1H), 7.37 - 7.30 (m, 2H), 6.70 (d, J = 8.4 Hz, 1H), 5.52 (m, 1H), 5.29 (d, J = 10.7 Hz, 1H), 4.90 (d, J = 10.7 Hz, 1H), 4.73 (q, J = 4.9 Hz, 1H), 4.10 (q, J = 6.3 Hz, 1H), 3.99 (m, 1H), 3.78 (t, J = 4.7 Hz, 4H), 3.75 - 3.70 (m, 1H), 3.69 (d, J = 3.4 Hz, 2H), 3.54 (s, 2H), 3.46 (m, 1H), 3.26 (s, 3H), 3.21 (d, J = 14.4 Hz, 1H), 3.10 (m, 1H), 2.78 (q, J = 5.9 Hz, 1H), 2.65 (t, J = 4.6 Hz, 4H), 2.58 (m, 1H), 2.42 (m, 1H), 2.31 (d, J = 14.3 Hz, 1H), 2.05 (m, 3H), 1.79 - 1.62 (m, 7H), 1.60 - 1.48 (m, 3H), 1.28 (s, 3H), 1.23 (d, J = 6.3 Hz, 3H), 1.01 (t, J = 7.4 Hz, 3H), 0.94 (t, J = 7.4 Hz, 3H).

[0410] Compound 3-P2 MS (ESI + m / z = 864.3 [M+H] +

[0411] 1H NMR (400 MHz, CDC13) δ 8.81 (d, J = 2.1 Hz, 1H), 8.41 (d, J = 1.6 Hz, 1H), 7.67 (d, J = 2.2 Hz, 1H), 7.61 (m, 1H), 7.36 (d, J = 8.6 Hz, 1H), 7.31 (s, 1H), 6.61 (d, J = 8.6 Hz, 1H), 5.59 (m, 1H), 5.31 (d, J = 10.7 Hz, 1H), 4.86 (d, J = 10.8 Hz, 1H), 4.73 (q, J = 4.9 Hz, 1H), 4.34 (q, J = 6.0 Hz, 1H), 4.19 (m, 2H), 3.78 (t, J = 4.6 Hz, 4H), 3.71 (d, J = 10.9 Hz, 1H), 3.62 (d, J = 10.9 Hz, 1H), 3.54 (s, 2H), 3.46 (m, 1H), 3.18 - 3.09 (m, 2H), 2.75 (q, J = 5.9 Hz, 1H), 2.66 (t, J = 4.7 Hz, 4H), 2.58 (m, 1H), 2.42 (m, 2H), 2.11 (m, 1H), 2.04 (m, 1H), 1.78 - 1.63 (m, 8H), 1.62 - 1.49 (m, 8H), 1.43 (d, J = 6.1 Hz, 3H), 1.01 (t, J = 7.4 Hz, 3H), 0.95 - 0.91 (m, 4H).

[0412] Synthesis of compound 4 of example 4

[0413] Compound 3-12 (20.0 mg, 26 μmol) and N,N-diisopropylethylamine (23.6 μL, 135.0 μmol) were added to a solution of compound 2-3 (9.0 mg, 52 μmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (20.0 mg, 52 μmol) in N,N-dimethylformamide (1.0 mL) successively, and stirred at room temperature for 3 hours. After the reaction was completed, compound 4 (3.0 mg, 3.2 μmol, yield: 15%) was obtained after purification by a reverse phase column (chromatography column: Boston Prime C18; 150*30 millimeters*5 microns; mobile phase A: H2O-(NH3H2O), NH3H2O concentration was 0.05%; mobile phase B: MeCN; MeCN ratio 50%-80%).

[0414] MS m / z (ESI): 919.4 [M+H] + .

[0415] 1H NMR (400 MHz, CDC13) δ 8.81 (d, J = 2.1 Hz, 1H), 8.43 (d, J = 1.6 Hz, 1H), 8.20 (d, J = 9.3 Hz, 1H), 7.67 (d, J = 2.1 Hz, 1H), 7.62 (dd, J = 8.6, 1.6 Hz, 1H), 7.36 (d, J = 8.6 Hz, 1H), 7.33 (s, 1H), 5.65 - 5.58 (m, 1H), 5.33 (d, J = 10.7 Hz, 1H), 5.10 - 5.04 (m, 1H), 4.89 - 4.84 (m, 1H), 4.77 - 4.71 (m, 1H), 4.41 - 4.30 (m, 2H), 4.25 - 4.15 (m, 2H), 3.82 - 3.76 (m, 4H), 3.74 - 3.70 (m, 1H), 3.62 (d, J = 10.9 Hz, 1H), 3.54 (s, 2H), 3.52 - 3.45 (m, 1H), 3.39 (s, 3H), 3.29 - 3.21 (m, 1H), 3.18 - 3.11 (m, 1H), 2.77 - 2.72 (m, 1H), 2.71 - 2.62 (m, 4H), 2.60 - 2.54 (m, 1H), 2.49 - 2.39 (m, 2H), 2.25 - 2.17 (m, 1H), 2.15 - 2.06 (m, 2H), 1.71 - 1.69 (m, 1H), 1.56 - 1.53 (m, 1H), 1.44 (d, J = 6.1 Hz, 3H), 1.26 - 1.23 (m, 3H), 1.12 (d, J = 6.4 Hz, 3H), 1.00 - 0.90 (m, 6H), 0.40 (s, 3H).

[0416] Synthesis of compound 5 of example 5

[0417] Compound Int-4 (23.0 mg, 31.04 μmol) and N,N-diisopropylethylamine (23.5 μL, 135.0 μmol) were added sequentially to a solution of 4-fluoro-2-(2-fluoroethyl)butyric acid 5A (6.2 mg, 40.6 μmol) and (2-oxime-cyanoethyl acetate)-N,N-dimethyl-morpholinourea hexafluorophosphate (23 mg, 54.0 μmol) in N,N-dimethylformamide (1.0 mL), and stirred at room temperature for 1 hour. Extracted with ethyl acetate (5 mL) and water (5 mL), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase column chromatography (Boston Prime C18 column; 150*30 mm*5 μm; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O ​​concentration 0.05%, NH4HCO3 concentration 2 mM; mobile phase B: MeCN; MeCN ratio 40%-80%) to obtain compound 5 (6 mg, 6.8 μmol, yield: 22%).

[0418] MS m / z (ESI): 875.4 [M+H] + .

[0419] 1H NMR (400 MHz, DMSO-d6) δ 8.62 (d, J = 8.4 Hz, 1H), 8.44 (d, J = 2.9 Hz, 1H), 8.41 (d, J = 1.6 Hz, 1H), 7.85 (s, 1H), 7.74 (dd, J = 8.9, 1.8 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.22 (d, J = 2.9 Hz, 1H), 5.95 (d, J = 11.1 Hz, 1H), 5.27 (t, J = 7.8 Hz, 1H), 4.73 (d, J = 11.0 Hz, 1H), 4.57 - 4.47 (m, 3H), 4.42 (s, 1H), 4.33 - 4.26 (m, 1H), 4.20 - 4.11 (m, 2H), 3.61 - 3.49 (m, 2H), 3.28 - 3.25 (m, 4H), 3.22 (s, 3H), 2.95 (d, J = 14.4 Hz, 1H), 2.71 - 2.63 (m, 2H), 2.48 - 2.44 (m, 4H), 2.38 - 2.31 (m, 1H), 2.22 (s, 3H), 2.19 - 2.13 (m, 1H), 2.06 - 1.94 (m, 1H), 1.92 - 1.80 (m, 3H), 1.66 - 1.59 (m, 1H), 1.40 - 1.32 (m, 5H), 1.28 - 1.18 (m, 3H), 0.94 - 0.86 (m, 6H), 0.32 (s, 2H).

[0420] Synthesis of compound 6 of Example 6

[0421] First Step: Synthesis of compound 6-1

[0422] Compound D8 (40.0 mg, 48.37 μmol), (1-ethoxycyclopropyl)trimethylsilane (16.9 mg, 96.73 μmol), sodium cyanoborohydride (9.1 mg, 145.10 μmol) and acetic acid (5.8 mg, 96.73 μmol) were added into isopropanol (1 mL), the mixture was stirred at 50 °C overnight, after the reaction was completed, the reaction solution was spin dried, the residue was purified by silica gel column chromatography (dichloromethane:methanol = 30:1) to give compound 6-1 (20.0 mg, 23.07 μmol, 47.5% yield).

[0423] MS: m / z (ESI): 867.5 [M+H] + .

[0424] Second Step: Synthesis of compound 6-2

[0425] Compound 6-1 (20.0 mg, 23.07 μmol) was dissolved in hydrochloric acid dioxane solution (4 M, 2 mL), the mixture was stirred at room temperature for 2 hours, the reaction conversion was monitored by LC-MS. The reaction solution was directly spin-dried to obtain compound 6-2 (15.0 mg, 19.5 μmol, yield 84.5%).

[0426] MS: m / z (ESI): 767.4 [M+H] + .

[0427] Step 3: synthesis of compound 6

[0428] Compound 6-2 (12.0 mg, 15.65 μmol), N,N-diisopropylethylamine (10.1 mg, 78.23 μmol, 13.6 μL) were added into a solution of 2-ethylbutyric acid 1A (3.6 mg, 31.29 μmol) and (2-oximino-cyanoacetic acid ethyl ester)-N,N-dimethyl-morpholinyl urea hexafluorophosphate (20.1 mg, 46.94 μmol) in N,N-dimethylformamide (1.0 mL) successively, and stirred at room temperature for 1 hour. Ethyl acetate (5 mL) and water (5 mL) were added for extraction, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by reversed-phase column (chromatography column: Boston Prime C18; 150*30 millimeter*5 microns; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration was 0.05%, NH4HCO3 concentration was 2 mM; mobile phase B: MeCN; MeCN ratio 30%-70%) to obtain compound 6 (6 mg, 6.89 μmol, yield: 44%). MS (ESI + m / z = 865.4 [M+H] + .

[0429] 1H NMR (400 MHz, DMSO-d6) δ 8.44 (d, J = 2.9 Hz, 1H), 8.41 (d, J = 1.5 Hz, 1H), 8.33 (d, J = 8.7 Hz, 1H), 7.83 (s, 1H), 7.73 (dd, J = 8.6, 1.6 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.21 (d, J = 2.9 Hz, 1H), 5.92 (d, J = 11.1 Hz, 1H), 5.37 - 5.29 (m, 2H), 4.73 (d, J = 11.1 Hz, 1H), 4.56 - 4.44 (m, 1H), 4.34 - 4.23 (m, 1H), 4.23 - 4.10 (m, 2H), 3.55 - 3.52 (m, 2H), 3.22 - 3.21 (m, 3H), 2.94 (d, J = 14.4 Hz, 1H), 2.69 - 2.66 (m, 4H), 2.36 - 2.31 (m, 1H), 2.20 - 2.10 (m, 2H), 2.03 - 1.94 (m, 1H), 1.70 - 1.59 (m, 2H), 1.56 - 1.35 (m, 6H), 1.33 (d, J = 6.1 Hz, 4H), 0.96 - 0.81 (m, 15H), 0.47 - 0.40 (m, 2H), 0.37 - 0.27 (m, 5H).

[0430] Synthesis of compound 7 of example 7

[0431] Compound 6-2 (10.0 mg, 12.45 μmol), N,N-diisopropylethylamine (8.0 mg, 62.23 μmol, 10.84 μL) were added into a solution of 2,3-dimethylbutanoic acid (2.9 mg, 24.89 μmol) and (2-hydroxyimino-cyanoacetic acid ethyl ester)-N,N-dimethyl-morpholinyl urea hexafluorophosphate (16.0 mg, 37.34 μmol) in N,N-dimethylformamide (1.0 mL) successively, stirred at room temperature for 2 hours. Ethyl acetate (5 mL) and water (5 mL) were added to extract, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase column (Boston Prime C18; 150*30 millimeter*5 microns; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration was 0.05%, NH4HCO3 concentration was 2 mM; mobile phase B: MeCN; MeCN ratio 30%-80%) to obtain compound 7 (1.6 mg, 1.87 μmol, yield: 15%).

[0432] MS (ESI + )m / z = 865.4 [M+H]+ .

[0433] 1 H NMR (400 MHz, DMSO-d6) δ 8.44 (d, J = 2.9 Hz, 1H), 8.41 (d, J = 5.8 Hz, 1H), 8.30 - 8.23 (m, 1H), 7.83 (d, J = 5.2 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.21 (d, J = 2.8 Hz, 1H), 5.99 - 5.89 (m, 1H), 5.36 - 5.27 (m, 1H), 4.72 (d, J = 10.9 Hz, 1H), 4.53 - 4.47 (m, 1H), 4.22 - 4.10 (m, 2H), 3.56 - 3.51 (m, 2H), 3.22 - 3.20 (m, 8H), 2.98 - 2.89 (m, 1H), 2.74 - 2.61 (m, 6H), 2.35 - 2.29 (m, 1H), 1.69 - 1.60 (m, 1H), 1.33 (d, J = 6.1 Hz, 3H), 1.28 - 1.21 (m, 5H), 1.07 (d, J = 7.0 Hz, 2H), 1.01 - 0.94 (m, 3H), 0.93 - 0.81 (m, 12H), 0.48 - 0.40 (m, 2H), 0.36 - 0.28 (m, 5H).

[0434] Synthesis of compound 8 of example 8

[0435] Compound 6-2 (47.7 mg, 62.23 μmol), N,N-diisopropylethylamine (40.2 mg, 311.16 μmol, 54.2 μL) were added into a solution of compound 4-fluoro-2-(2-fluoroethyl)butanoic acid 5A (18.9 mg, 124.46 μmol) and (2-oximino-cyanoacetic acid ethyl ester)-N,N-dimethyl-morpholinyl urea hexafluorophosphate (COMU) (80.0 mg, 186.69 μmol) in N,N-dimethylformamide (1.0 mL) successively, stirred at room temperature for 4 hours. Ethyl acetate (5 mL) and water (5 mL) were added to extract, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase column (Boston Prime C18; 150*30 millimeter*5 microns; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration was 0.05%, NH4HCO3 concentration was 2 mM; mobile phase B: MeCN; MeCN ratio 35%-85%) to obtain compound 8 (14 mg, 15.53 μmol, yield: 25%).

[0436] MS m / z (ESI): 901.4 [M+H] + .

[0437] 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (d, J = 8.5 Hz, 1H), 8.44 (d, J = 2.9 Hz, 1H), 8.41 (d, J = 1.6 Hz, 1H), 7.84 (s, 1H), 7.73 (dd, J = 8.7, 1.7 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.21 (d, J = 2.9 Hz, 1H), 5.95 (d, J = 11.1 Hz, 1H), 5.27 (t, J = 7.7 Hz, 1H), 4.72 (d, J = 11.0 Hz, 1H), 4.57 - 4.48 (m, 3H), 4.44 - 4.39 (m, 1H), 4.33 - 4.24 (m, 1H), 4.19 - 4.10 (m, 2H), 3.59 - 3.50 (m, 2H), 3.24 - 3.20 (m, 7H), 2.98 - 2.90 (m, 1H), 2.73 - 2.61 (m, 7H), 2.38 - 2.29 (m, 1H), 2.21 - 2.12 (m, 1H), 2.05 - 1.94 (m, 1H), 1.94 - 1.78 (m, 4H), 1.69 - 1.58 (m, 2H), 1.38 - 1.30 (m, 4H), 1.27 - 1.19 (m, 2H), 0.93 - 0.85 (m, 6H), 0.47 - 0.40 (m, 2H), 0.34 - 0.30 (m, 4H).

[0438] Synthesis of compound 9

[0439] First Step: Synthesis of compound 9-1

[0440] Compound D8 (40.0 mg, 48.37 μmol), 3-oxetanone (7.0 mg, 97 μmol), sodium cyanoborohydride (6.0 mg, 97 μmol) and acetic acid (9.0 mg, 145 μmol) were added into isopropanol (3 mL), the mixture was reacted at room temperature for 2 hours, after the reaction was completed, the reaction solution was spin dried, the residue was purified by silica gel column chromatography (dichloromethane:methanol = 30:1) to obtain compound 9-1 (38.0 mg, 43 μmol, yield 89%).

[0441] MS: m / z (ESI): 883.4 [M+H] + .

[0442] Second Step: Synthesis of compound 9-2

[0443] Compound 9-1 (38.0 mg, 43 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added dropwise to the solution at 0 °C, the mixture was stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, the residue was poured into saturated sodium bicarbonate solution (20 mL), extracted with dichloromethane (10 ml x 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to obtain compound 9-2 (32.0 mg, 40.85 μmol, yield 95%).

[0444] MS: m / z (ESI): 783.4 [M+H] + .

[0445] Step 3: synthesis of compound 9

[0446] Compound 9-2 (40.0 mg, 51 μmol), 2-ethylbutyric acid (9.0 mg, 76 μmol), triethylamine (10.0 mg, 98.8 μmol) and 2-(7-azobenzo triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (39.0 mg, 102 μmol) were dissolved in N,N-dimethylformamide (2 mL), the mixture was reacted at room temperature overnight, the reaction solution was poured into water (10 mL), extracted with ethyl acetate (10 mL x 3), the combined organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated, and the residue was purified by reversed-phase column (the chromatographic column was Boston Prime C18; 150*30 mm*5 μm; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration was 0.05%, NH4HCO3 concentration was 2 mM; mobile phase B: MeCN; MeCN ratio 30%-80%) to obtain compound 9 (10.0 mg, 11.22 μmol, yield: 22%).

[0447] MS (ESI + )m / z = 881.5 [M+H] + .

[0448] 1H NMR (400 MHz, CDC13) δ 8.49 (d, J = 2.9 Hz, 1H), 8.39 (d, J = 1.6 Hz, 1H), 7.60 (dd, J = 8.6, 1.6 Hz, 1H), 7.35 (d, J = 8.6 Hz, 1H), 7.30 (s, 1H), 7.08 (d, J = 3.0 Hz, 1H), 6.62 (d, J = 8.6 Hz, 1H), 5.64 - 5.56 (m, 1H), 5.32 (d, J = 10.8 Hz, 1H), 4.84 (d, J = 10.7 Hz, 1H), 4.76 - 4.73 (m, 1H), 4.73 - 4.68 (m, 2H), 4.65 (t, J = 6.2 Hz, 2H), 4.32 - 4.23 (m, 2H), 4.20 - 4.12 (m, 1H), 3.72 (d, 1H), 3.62 (d, J = 10.7 Hz, 1H), 3.60 - 3.55 (m, 1H), 3.48 - 3.43 (m, 1H), 3.35 (s, 3H), 3.34 - 3.28 (m, 4H), 3.16 - 3.12 (m, 1H), 3.12 - 3.08 (m, 1H), 2.78 - 2.71 (m, 1H), 2.58 - 2.52 (m, 4H), 2.49 - 2.38 (m, 2H), 2.16 - 2.09 (m, 1H), 2.08 - 2.02 (m, 1H), 2.01 - 1.94 (m, 1H), 1.62 - 1.59 (m, 1H), 1.56 - 1.50 (m, 2H), 1.43 (d, J = 6.1 Hz, 3H), 1.01 (t, J = 7.4 Hz, 3H), 0.98 - 0.96 (m, 2H), 0.96 - 0.94 (m, 5H), 0.94 - 0.93 (m, 2H), 0.93 - 0.91 (m, 1H), 0.41 (s, 3H).

[0449] Synthesis of compound 10 of example 10

[0450] First step: synthesis of compound 10-1

[0451] Compound D8 (50.0 mg, 60.46 μmol), N-methyl-4-piperidone (13.7 mg, 120.91 μmol), sodium cyanoborohydride (11.4 mg, 181.37 μmol), acetic acid (7.3 mg, 120.91 μmol, 7.0 μL) were added into isopropanol (2 mL). The reaction was stirred at 50 °C overnight. After monitoring the reaction was finished by LC-MS, the reaction was spin dried, the residue was purified by silica gel column chromatography (dichloromethane:methanol = 30:1) to give compound 10-1 (29.0 mg, 31.38 μmol, 52% yield).

[0452] MS: m / z (ESI): 924.5 [M+H] + .

[0453] Second Step: Synthesis of compound 10-2

[0454] Compound 10-1 (29.0 mg, 31.38 µmol) was added to a solution of hydrochloric acid dioxane (4 M, 5 mL). The reaction was stirred at room temperature. After the reaction was monitored to be completed by LC-MS, the reaction was directly spin dried to give compound 10-2 (25.0 mg, 30.34 µmol, yield 97%).

[0455] MS: m / z (ESI): 824.4 [M+H] + .

[0456] Third Step: Synthesis of compound 10

[0457] Compound 10-2 (25.0 mg, 30.34 µmol), 2-ethylbutyric acid (7.1 mg, 60.67 µmol), 2-(7-azobenzo-triazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (33.1 mg, 87.15 µmol) and N,N-diisopropyl ethylamine (18.8 mg, 145.26 µmol, 25.30 µL) were added to N,N-dimethylformamide (2 mL). The reaction was stirred at room temperature for 2 hours. The reaction was poured into water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phase was combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated, and the residue was purified by reverse phase column (the chromatographic column was Boston Prime C18; 150*30 millimeters*5 microns; mobile phase A: H2O-(NH3H2O-NH4HCO3), NH3H2O concentration was 0.05%, NH4HCO3 concentration was 2 mM; mobile phase B: MeCN; MeCN ratio 30%-70%) to give compound 10 (5.3 mg, 5.76 µmol, yield: 19%).

[0458] MS: m / z (ESI): 922.5 [M+H] + .

[0459] 1H NMR (400 MHz, DMSO-d6) δ 8.43 (d, J = 2.9 Hz, 1H), 8.41 (d, J = 1.6 Hz, 1H), 8.32 (d, J = 8.7 Hz, 1H), 7.82 (s, 1H), 7.72 (dd, J = 9.0, 1.3 Hz, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.19 (d, J = 2.9 Hz, 1H), 5.91 (d, J = 11.1 Hz, 1H), 5.33 (t, J = 7.8 Hz, 1H), 4.73 (d, J = 11.1 Hz, 1H), 4.54 - 4.47 (m, 1H), 4.35 - 4.23 (m, 1H), 4.20 - 4.11 (m, 2H), 3.56 - 3.52 (m, 2H), 3.25 - 3.23 (m, 4H), 3.21 (s, 3H), 2.97 - 2.91 (m, 1H), 2.86 - 2.80 (m, 2H), 2.69 - 2.65 (m, 1H), 2.65 - 2.59 (m, 5H), 2.48 - 2.45 (m, 1H), 2.36 - 2.32 (m, 1H), 2.20 - 2.12 (m, 6H), 1.97 - 1.86 (m, 2H), 1.77 - 1.72 (m, 2H), 1.63 - 1.57 (m, 1H), 1.52 - 1.36 (m, 6H), 1.33 (d, J = 6.1 Hz, 3H), 0.95 - 0.91 (m, 3H), 0.91 - 0.89 (m, 5H), 0.89 - 0.87 (m, 2H), 0.87 - 0.83 (m, 3H), 0.31 (s, 3H).

[0460] Biological test

[0461] Test Example 1, Effect of Compound on Proliferative Activity of Tumor Cells

[0462] Experimental materials and instruments:

[0463] The materials required for this experiment include: cell culture medium RPMI-1640 (BasalMedia, #L240KJ); fetal bovine serum (FBS) (Proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 0.25% trypsin (Gibco, #25200-072); 100% DMSO (Sigma, #D2650); 96-well sterile transwell culture plate (Corning, #3599); 96-well plate (Corning, #3610); CellTiter-Glo® Luminescent Cell Viability Assay Kit (Promega, #G7571); and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Sigma, #M5655). 2.0 Luminescent Cell Viability Assay Kit (Vazyme, #DD1101); 25 mL pipette (Corning); 5 mL pipette (Corning); P1000 pipette tips, P200 pipette tips and P10 pipette tips (Axygen).

[0464] The instruments and equipment required for this experiment include: Eppendorf pipette; Eppendorf pipette gun; Eppendorf centrifuge; constant temperature carbon dioxide incubator (ThermoFisher); automatic cell counter Vi-cell XR (Beckman Coulter); Envision enzyme labeler (Perkin Elmer).

[0465] The cells required for this experiment include: KRAS G12D Mutant cell strain AsPC-1 (ATCC, #CRL-1682TM) is cultured in RPMI-1640 medium containing 10% FBS.

[0466] Experimental method:

[0467] After rinsing the cells with PBS phosphate buffer, the AsPC-1 cells are digested from the culture bottle using 0.25% trypsin and resuspended with the corresponding fresh complete culture medium. After counting, the AsPC-1 cell density is adjusted to 2000 cells / 90 μL / well, and 90 μL is taken and added to a 96-well plate, which is placed in a 37°C, 5% carbon dioxide cell incubator for overnight culture. Compound stock solution 10 mM (solvent is DMSO), 10-fold dilution to 1 mM using DMSO, then 100-fold dilution to 10 μM in a 96-well transparent sterile culture plate using complete culture medium, and then 1% DMSO-containing complete culture medium is used for 3-fold gradient dilution, with 9 concentration gradients for continuous dilution; Take 10 μL / well of gradient-diluted compound and add it to the cell culture well, so that the final DMSO content of each well is one thousandth. The positive control group is the culture medium well without planting cells; the negative control group is the cell-containing compound-free well. The cell plate is incubated at 37°C, 5% carbon dioxide for 5 days. Take out the cell plate and add an equal volume of CellCounting-Lite 2.0 detection reagent to each well, shake well for 2-5 min to fully lyse the cells, and place at room temperature for 10 min to stabilize the luminescence signal. The Envision enzyme labeler is used to read the luminescence value. According to the following formula, the inhibition rate is calculated: Inhibition% = (Signal negative control–Signal sample ) / (Signal negative control–Signal positive control)*100. IC was calculated using IDBS XLfit with 4-parameter fitting. 50 Numerical value. Measured IC 50 The values ​​are shown in Table 1.

[0468] Table 1

[0469] Test Example 2: Pan-RAS(ON) Inhibitor Combined with FAK Inhibitor on KRAS G12C Effect of mutation on the inhibition of human pancreatic cancer MiaPaCa-2 cell proliferation

[0470] Experimental materials and instruments:

[0471] The reagents and consumables required for this experiment include: DMEM cell culture medium (BasalMedia, #L110KJ); fetal bovine serum (FBS) (proteintech, #PM00011); horse serum (HS) (Gibco, #16050-130); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile culture plate with permeable bottom (Corning, #3599); 96-well plate (Corning, #3610); ifebemtinib (MCE, #HY-122844); defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ). Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0472] The equipment required for this experiment includes: Eppendorf pipettes, Eppendorf pipettes, a constant temperature carbon dioxide incubator (ThermoFisher), a Countstar automated cell counter, and an Envision microplate reader (PerkinElmer).

[0473] The cells required for this experiment include: MiaPaCa-2 (ATCC, #CRL-1420), and the complete culture medium is DMEM medium containing 10% FBS and 2.5% HS.

[0474] Experimental methods:

[0475] PBS phosphate buffered saline was used to rinse the cells, then 0.25% trypsin was used to digest the MiaPaCa-2 cells, after resuspension in complete medium, the cells were counted and adjusted to a cell density of 700 cells / 80 μL / well, then plated into a 96-well plate and incubated in a constant temperature carbon dioxide incubator overnight. Compound PA-1 (synthesized according to WO2024067857A1 Example 6) was prepared into a 100 μM solution with DMSO, FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917) and Narmafotinib (MCE, #HY-145652) were prepared into 3000 μM solutions respectively; the prepared compounds were diluted 100 times in complete medium in a 96-well sterile transwell culture plate, and the concentrations of compound PA-1 and FAK inhibitors were 1 μM and 30 μM respectively; compound PA-1 was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 7 concentration gradients were continuously diluted; Ifebemtinib, Defactinib, VS-4718 and Narmafotinib were diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted compound PA-1 and FAK inhibitors were transferred to the MiaPaCa-2 cell culture plate plated the day before in an orthogonal manner. The positive control group was a well containing 0.2% DMSO complete medium without cells; the negative control group was a well containing 0.2% DMSO complete medium with cells. The cell plate was placed in a 37°C, CO2 cell incubator for 7 days. The cell plate was removed, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, shaken for 2-5 minutes to fully lyse the cells, and placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100×(average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model (Bliss Independence model) was used for statistical analysis to determine the effect (synergy, independence or antagonism) between drugs (Bliss, 1939). The Bliss independent model assumes a random process of two drugs acting independently, and the expected joint effect can be calculated according to the probability of independent events. The theoretical curve of the expected effect of combination is calculated by the following equation:

[0476] Bliss effect = effect A + effect B - effect A x effect B

[0477] Wherein: Effect A and Effect B are the effects of drugs A and B individually at specific concentrations. A synergistic score greater than 10 points is considered synergistic, -10-10 points is additive, and less than -10 points is antagonistic.

[0478] Experimental results: As shown in Figure 1, the combination of compound PA-1 with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed strong synergistic inhibition of KRAS G12C mutant human pancreatic cancer MiaPaCa-2 cells showed strong synergistic inhibition of proliferation.

[0479] Test Example 3, pan-RAS (ON) inhibitor combined with FAK inhibitor on KRAS G12D mutant human colon cancer GP2d cell proliferation inhibition

[0480] Experimental materials and instruments:

[0481] The reagent consumables required in this experiment include: cell culture medium DMEM (BasalMedia, #L110KJ); fetal bovine serum (FBS) (proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile culture plate (Corning, #3599); 96-well plate (Corning, #3610); RMC-6236 (MCE, #HY-148439); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0482] The instruments and equipment required in the experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme label instrument (PerkinElmer).

[0483] The cells required in the experiment include: GP2d (COBIOER, #CBP60010), and the complete culture medium is DMEM medium containing 10% FBS.

[0484] Experimental method:

[0485] PBS phosphate buffer solution was used to rinse the cells, and then the GP2d cells were digested with 0.25% trypsin. After resuspension in complete medium, the cells were counted and adjusted to a cell density of 2000 cells / 80 μL / well, and then plated into a 96-well plate and cultured overnight in a constant temperature carbon dioxide incubator. The pan-RAS(ON) inhibitor RMC-6236 (MCE, #HY-148439) or compound PA-1 (synthesized according to WO2024067857A1 Example 6) was respectively prepared into a 100 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917) and Narmafotinib (MCE, #HY-145652) were respectively prepared into a 3000 μM solution; the prepared compounds were diluted 100 times with complete medium in a 96-well transwell sterile culture plate, and the concentrations of the pan-RAS(ON) inhibitor and the FAK inhibitor were 1 μM and 30 μM respectively; the compound RMC-6236 or the compound PA-1 was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 7 concentration gradients were continuously diluted; the FAK inhibitor was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitor and FAK inhibitor were respectively transferred to the GP2d cell culture plate plated the day before in an orthogonal manner. The positive control group was a well of complete medium containing 0.2% DMSO without cells; the negative control group was a well of complete medium containing 0.2% DMSO with cells. The cell plate was placed in a 37°C, CO2cell incubator for culture for 7 days. The cell plate was taken out, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, and the cells were shaken and mixed for 2-5 minutes to fully lyse the cells. The plate was placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100 x (average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect (synergy, independence or antagonism) between drugs.

[0486] Experimental results: As shown in FIG. 2, the combination of compound RMC-6236 and the FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed a synergistic effect on KRAS G12DThe mutant human colon cancer GP2d cell proliferation was strongly synergistically inhibited; as shown in Figure 3, the compound PA-1 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib also showed strong synergistic inhibition on the cell line.

[0487] Test Example 4, pan-RAS (ON) inhibitor combined with FAK inhibitor on KRAS G12D Effect of inhibition of proliferation of mutant human pancreatic cancer AsPC-1 cells

[0488] Experimental materials and instruments:

[0489] The reagent consumables required in this experiment include: cell culture medium RPMI-1640 (BasalMedia, #L240KJ); fetal bovine serum (FBS) (proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile transwell culture plate (Corning, #3599); 96-well plate (Corning, #3610); RMC-6236 (MCE, #HY-148439); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0490] The instruments and equipment required in this experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme label instrument (PerkinElmer).

[0491] The cells required in this experiment include: AsPC-1 (ATCC, #CRL-1682TM), and the complete culture medium is RPMI-1640 culture medium containing 10% FBS.

[0492] Experimental Methods:

[0493] After rinsing the cells with PBS phosphate buffer, AsPC-1 cells were digested with 0.25% trypsin, resuspended with complete medium and counted. The cell density was adjusted to 2000 cells / 80 μL / well, and plated into 96-well plates, which were incubated in a constant temperature carbon dioxide incubator overnight. The pan-RAS(ON) inhibitor RMC-6236 (MCE, #HY-148439) or compound PA-1 (synthesized according to WO2024067857A1 Example 6) was respectively prepared into a 100 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917) or Narmafotinib (MCE, #HY-145652) were respectively prepared into a 3000 μM solution; the diluted compounds were diluted 100 times with complete medium in a 96-well transwell sterile culture plate, and the concentrations of the pan-RAS(ON) inhibitor and the FAK inhibitor were 1 μM and 30 μM, respectively; RMC-6236 or compound PA-1 was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 6-7 concentration gradients were continuously diluted; the FAK inhibitor was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitor and FAK inhibitor were transferred into the AsPC-1 cell culture plate plated the day before in an orthogonal manner. The positive control group was a well of complete medium containing 0.2% DMSO without cells; the negative control group was a well of complete medium containing 0.2% DMSO treated with cells. The cell plate was placed in a 37°C, CO2cell incubator for 7 days. The cell plate was taken out, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, and the cells were shaken and mixed for 2-5 minutes to fully lyse the cells. The plate was placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100 x (average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect (synergy, independence or antagonism) between drugs.

[0494] Experimental results: As shown in Figure 4, compound RMC-6236 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed synergistic inhibition on the proliferation of KRAS G12D mutant human pancreatic cancer AsPC-1 cells; as shown in Figure 5, compound PA-1 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib also showed synergistic inhibition on the cell line.

[0495] Test Example 5, pan-RAS (ON) inhibitor combined with FAK inhibitor on KRAS G12V mutant human non-small cell lung cancer H441 proliferation inhibition

[0496] Experimental materials and instruments:

[0497] The reagent consumables required for this experiment include: cell culture medium RPMI-1640 (BasalMedia, #L240KJ); fetal bovine serum (FBS) (proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile transwell culture plate (Corning, #3599); 96-well plate (Corning, #3610); RMC-6236 (MCE, #HY-148439); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0498] The instruments and equipment required for this experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme label instrument (PerkinElmer).

[0499] Cells required for this experiment include: H441 (ATCC, #HTB-174), complete culture medium is RPMI-1640 medium containing 10% FBS.

[0500] Experimental method:

[0501] After rinsing the cells with PBS phosphate buffer, H441 cells were digested with 0.25% trypsin, resuspended with complete culture medium and counted. The cell density was adjusted to 1500 cells / 80 μL / well, plated into a 96-well plate and incubated in a constant temperature carbon dioxide incubator overnight. The pan-RAS(ON) inhibitor RMC-6236 (MCE, #HY-148439) or compound PA-1 (synthesized according to WO2024067857A1 Example 6) was respectively prepared into a 100 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917) or Narmafotinib (MCE, #HY-145652) were respectively prepared into a 3000 μM solution; the diluted compounds were diluted 100 times with complete culture medium in a 96-well transwell sterile culture plate, and the concentrations of pan-RAS(ON) inhibitors and FAK inhibitors were 1 μM and 30 μM respectively; compound RMC-6236 or compound PA-1 was diluted by 3 times gradient with the prepared complete culture medium containing 1% DMSO, and 5-7 concentration gradients were continuously diluted; the FAK inhibitors were diluted by 3 times gradient with the prepared complete culture medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitors and FAK inhibitors were transferred to the H441 cell culture plate plated the day before in an orthogonal manner. The positive control group was a well of complete culture medium containing 0.2% DMSO without cells; the negative control group was a well of complete culture medium containing 0.2% DMSO treated with cells. The cell plate was placed in a 37°C, CO2 cell incubator for 7 days. The cell plate was taken out, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, shaken for 2-5 minutes to fully lyse the cells, and placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100 x (average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect between drugs (synergy, independence or antagonism).

[0502] Experimental results: As shown in Figure 6, the compound RMC-6236 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed strong synergistic inhibition on the proliferation of KRAS G12V The proliferation of human non-small cell lung cancer H441 cells was also strongly synergistically inhibited; as shown in Figure 7, the compound PA-1 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib also showed strong synergistic inhibition on this cell line.

[0503] Test Example 6, pan-RAS (ON) inhibitor combined with FAK inhibitor on KRAS G12S Effect on the proliferation inhibition of human non-small cell lung cancer A549 cells

[0504] Experimental materials and instruments:

[0505] The reagent consumables required for this experiment include: cell culture medium DMEM (BasalMedia, #L110KJ); fetal bovine serum (FBS) (proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile transwell culture plate (Corning, #3599); 96-well plate (Corning, #3610); RMC-6236 (MCE, #HY-148439); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0506] The instruments and equipment required for this experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme labeler (PerkinElmer).

[0507] Cells required for this experiment include: A549 (ATCC, # CRM-CCL-185), complete culture medium is DMEM medium containing 10% FBS.

[0508] Experimental method:

[0509] After rinsing the cells with PBS phosphate buffer, A549 cells were digested with 0.25% trypsin, resuspended with fresh complete culture medium, counted, and adjusted to a cell density of 300 cells / 80 μL / well, plated into a 96-well plate, and incubated in a constant-temperature carbon dioxide incubator overnight. The pan-RAS(ON) inhibitor RMC-6236 (MCE, # HY-148439) or compound PA-1 (synthesized according to WO2024067857A1 Example 6) was respectively prepared into a 100 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, # HY-122844), Defactinib (MCE, # HY-12289), VS-4718 (MCE, # HY-13917), or Narmafotinib (MCE, # HY-145652) were respectively prepared into a 3000 μM solution; the diluted compounds were diluted 100 times with complete culture medium in a 96-well transwell sterile culture plate, and the concentrations of the pan-RAS(ON) inhibitor and the FAK inhibitor were 1 μM and 30 μM, respectively; the compound RMC-6236 or the compound PA-1 was diluted by 3 times gradient with the prepared 1% DMSO culture solution as the starting concentration, and 7 concentration gradients were continuously diluted; the FAK inhibitor was diluted by 3 times gradient with the prepared 1% DMSO culture solution as the starting concentration, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitor and FAK inhibitor were transferred to the A549 cell culture plate plated the day before in an orthogonal manner. The positive control group was a well containing 0.2% DMSO complete culture medium without cells; the negative control group was a well containing 0.2% DMSO complete culture medium treated with cells. The cell plate was placed in a 37°C, CO2cell incubator for 7 days. The cell plate was taken out, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, shaken and mixed for 2-5 minutes to fully lyse the cells, and the light emission signal was stabilized at room temperature for 10 minutes. The Envision enzyme labeler was used to read the light emission value. Cell survival rate (%) = 100-100 x (average value of light emission value of negative control well - light emission value of compound well) / (average value of light emission value of negative control - average value of light emission value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect (synergy, independence, or antagonism) between drugs.

[0510] Experimental results: As shown in Figure 8, the compound RMC-6236 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed strong synergistic inhibition on the proliferation of KRAS wild-type cell H1975; as shown in Figure 9, the compound PA-1 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib also showed strong synergistic inhibition on the proliferation of KRAS wild-type cell H1975. G12S Experimental results: As shown in Figure 8, the compound RMC-6236 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed strong synergistic inhibition on the proliferation of KRAS wild-type cell H1975; as shown in Figure 9, the compound PA-1 combined with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib also showed strong synergistic inhibition on the proliferation of KRAS wild-type cell H1975.

[0511] Test Example 7, Effect of Pan-RAS (ON) Inhibitor Combined with FAK Inhibitor on Inhibition of Proliferation of KRAS Wild-Type Cell H1975

[0512] Experimental materials and instruments:

[0513] The reagent consumables required in this experiment include: cell culture medium RPMI-1640 (BasalMedia, #L240KJ); fetal bovine serum (FBS) (proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile transwell culture plate (Corning, #3599); 96-well plate (Corning, #3610); RMC-6236 (MCE, #HY-148439); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0514] The instruments and equipment required in this experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme label instrument (PerkinElmer).

[0515] Cells required for this experiment include: H1975 (ATCC, #CRL-5908) and complete culture medium is RPMI-1640 medium with 10% FBS.

[0516] Experimental Methods:

[0517] PBS phosphate buffer solution was used to rinse the cells, and then the H1975 cells were digested with 0.25% trypsin. After resuspension in fresh complete culture medium, the cells were counted and adjusted to a cell density of 350 cells / 80 μL / well, and then plated into a 96-well plate and cultured overnight in a constant temperature carbon dioxide incubator. The pan-RAS(ON) inhibitor RMC-6236 (MCE, #HY-148439) or compound PA-1 (synthesized according to Example 6 of WO2024067857A1) was respectively prepared into a 900 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917) or Narmafotinib (MCE, #HY-145652) were respectively prepared into a 3000 μM solution; the diluted compounds were diluted 100 times with complete culture medium in a 96-well transwell sterile culture plate, and the concentrations of the pan-RAS(ON) inhibitor and the FAK inhibitor were 9 μM and 30 μM respectively; RMC-6236 or compound PA-1 (synthesized according to Example 6 of WO2024067857A1) was diluted by 3 times gradient with the prepared complete culture medium containing 1% DMSO, and 6-7 concentration gradients were continuously diluted; the FAK inhibitor was diluted by 3 times gradient with the prepared complete culture medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitor and FAK inhibitor were respectively transferred to the H1975 cell culture plate plated the day before in an orthogonal manner. The positive control group was a well of complete culture medium containing 0.2% DMSO without cells; the negative control group was a well of complete culture medium containing 0.2% DMSO treated with cells. The cell plate was placed in a 37°C, CO2cell incubator for culture for 7 days. The cell plate was taken out, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, and the cells were shaken and mixed for 2-5 minutes to fully lyse the cells. The plate was placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100 x (average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect (synergy, independence or antagonism) between drugs.

[0518] Experimental results: As shown in FIG. 10, the combination of compound RMC-6236 and the FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed a synergistic effect on the inhibition of KRAS WTThe proliferation of human non-small cell lung cancer H1975 cells was also synergistically inhibited. As shown in Figure 11, the combination of compound PA-1 with FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed synergistic inhibition of KRAS WT The proliferation of human non-small cell lung cancer H1975 cells was also synergistically inhibited.

[0519] Test Example 8, the effect of combination of pan-RAS (ON) inhibitor and FAK inhibitor on NRAS G13D The effect of combination of pan-RAS (ON) inhibitor and FAK inhibitor on NRAS

[0520] Experimental materials and instruments:

[0521] The reagent consumables required for this experiment include: cell culture medium RPMI-1640 (BasalMedia, #L240KJ); fetal bovine serum (FBS) (proteintech, #PM00011); PBS phosphate buffer (BasalMedia, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile transwell culture plate (Corning, #3599); 96-well plate (Corning, #3610); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (BasalMedia, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0522] The instruments and equipment required for this experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme labeler (PerkinElmer).

[0523] The cells required for this experiment include: H929 (ATCC, #CRL-3580), complete culture medium is RPMI-1640 culture medium containing 10% FBS.

[0524] Experimental method:

[0525] After rinsing the cells with PBS phosphate buffer, the H929 cells were digested with 0.25% trypsin, resuspended with complete medium, counted, and adjusted to a cell density of 4000 cells / 80 μL / well, plated into a 96-well plate, and incubated in a constant-temperature carbon dioxide incubator overnight. The pan-RAS(ON) inhibitor compound PA-1 (synthesized according to WO2024067857A1 Example 6) was prepared into a 160 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917), or Narmafotinib (MCE, #HY-145652) were prepared into 3000 μM solutions with DMSO, respectively; the diluted compounds were diluted 100 times with complete medium in a 96-well sterile transwell culture plate, and the concentrations of the pan-RAS(ON) inhibitor and the FAK inhibitor were 1.6 μM and 30 μM, respectively; the compound PA-1 was diluted by 2-fold gradient with the prepared complete medium containing 1% DMSO, and 7 concentration gradients were continuously diluted; the FAK inhibitor was diluted by 3-fold gradient with the prepared complete medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitor and FAK inhibitor were transferred to the H929 cell culture plate plated the previous day in an orthogonal manner. The positive control group was a well containing 0.2% DMSO complete medium without cells; the negative control group was a well containing 0.2% DMSO complete medium with cells. The cell plate was placed in a 37°C, CO2cell incubator for 7 days. The cell plate was removed, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, and the cells were shaken and mixed for 2-5 minutes to fully lyse the cells. The plate was placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100 x (average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect between drugs (synergy, independence, or antagonism)

[0526] Experimental results: As shown in FIG. 12, the combination of compound PA-1 and the FAK inhibitors Ifebemtinib, VS-4718, Defactinib, or Narmafotinib showed synergistic inhibition of the proliferation of NRAS G13D H929 cells showed synergistic inhibition of the proliferation of H929 cells.

[0527] Test Example 9, pan-RAS (ON) inhibitor in combination with FAK inhibitor on HRAS F82L Effect of human endometrial cancer AN3 CA cell proliferation inhibition

[0528] Experimental materials and instruments:

[0529] The reagent consumables required for this experiment include: cell culture medium MEM (Basal Media, #L570KJ); fetal bovine serum (FBS) (proteintech, #PM00011); MEM non-essential amino acid NEAA solution (100X) (Gibco, #11140050); 100mM sodium pyruvate (Gibco, #11360070); PBS phosphate buffer (Basal Media, #B320KJ); 100% DMSO (Sigma, #D2650); 96-well sterile culture plate with transparent bottom (Corning #3599); 96-well plate (Corning, #3610); RMC-6236 (MCE, #HY-148439); Ifebemtinib (MCE, #HY-122844); Defactinib (MCE, #HY-12289); VS-4718 (MCE, #HY-13917); Narmafotinib (MCE, #HY-145652); 0.25% trypsin (Basal Media, #S310KJ); Luminescent Cell Viability Detection Kit (VKEY-BIO, A2010003N); T75 cell culture flask (Corning), 25mL serum pipette tip (Corning), 5mL serum pipette tip (Corning), P1000 pipette tip and P200 pipette tip (Axygen).

[0530] The instruments and equipment required for this experiment include: Eppendorf pipette, Eppendorf pipette gun, constant temperature carbon dioxide incubator (ThermoFisher), Countstar automatic cell counter, Envision enzyme label instrument (PerkinElmer).

[0531] The cells required for this experiment include: AN3 CA (ATCC, #HTB-111), the complete culture medium is MEM + 10% FBS + 1% NEAA + 1mM sodium pyruvate

[0532] Experimental method:

[0533] PBS phosphate buffered saline was used to rinse the cells, then AN3 CA cells were digested with 0.25% trypsin, resuspended with complete medium and counted. The cell density was adjusted to 1500 cells / 80 μL / well, and plated into 96-well plates, which were incubated in a constant temperature carbon dioxide incubator overnight. The pan-RAS(ON) inhibitor RMC-6236 (MCE, #HY-148439) or compound PA-1 (synthesized according to WO2024067857A1 Example 6) was respectively prepared into a 1000 μM solution with DMSO, and the FAK inhibitors Ifebemtinib (MCE, #HY-122844), Defactinib (MCE, #HY-12289), VS-4718 (MCE, #HY-13917) or Narmafotinib (MCE, #HY-145652) were respectively prepared into a 3000 μM solution; the diluted compounds were diluted 100 times with complete medium in a 96-well transwell sterile culture plate, and the concentrations of the pan-RAS(ON) inhibitor and the FAK inhibitor were 10 μM and 30 μM respectively; RMC-6236 or compound PA-1 was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 6-7 concentration gradients were continuously diluted; the FAK inhibitor was diluted by 3 times gradient with the prepared complete medium containing 1% DMSO, and 6 concentration gradients were continuously diluted. Finally, 10 μL of the gradient-diluted pan-RAS(ON) inhibitor and FAK inhibitor were respectively transferred to the AN3 CA cell culture plate plated the day before in an orthogonal manner. The positive control group was a well of complete medium containing 0.2% DMSO without cells; the negative control group was a well of complete medium containing 0.2% DMSO treated with cells. The cell plate was placed in a 37°C, CO2cell incubator for 7 days. The cell plate was taken out, 50 μL of CTG reagent (VKEY-BIO, A2010003N) was added to each well, and the cells were fully lysed by shaking for 2-5 minutes. The plate was placed at room temperature for 10 minutes to stabilize the luminescence signal. The Envision enzyme labeler was used to read the luminescence value. Cell survival rate (%) = 100-100 x (average value of luminescence value of negative control well-luminescence value of compound well) / (average value of luminescence value of negative control- average value of luminescence value of positive control). The Combenefit software was used to analyze the survival rate of the matrix, and the Bliss independent model was used for statistical analysis to determine the effect between drugs (synergy, independence or antagonism)

[0534] Experimental results: As shown in Figure 13, the combination of compound RMC-6236 and FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib showed a synergistic effect on HRAS F82LThe proliferation of human endometrial cancer AN3CA cells was synergistically inhibited by the combination of the compounds PA-1 and FAK inhibitors. As shown in Figure 14, the combination of the compound PA-1 and FAK inhibitors Ifebemtinib, VS-4718, Defactinib or Narmafotinib also showed synergistic inhibition on the cell line.

Claims

A combination drug combination comprising a pan-RAS(ON) inhibitor and a FAK inhibitor. The combination drug combination of claim 1, wherein the FAK inhibitor is selected from Ifebemtinib (IN-10018, BI-853520), Defactinib (VS-6063, PF-04554878), Narmafotinib (AMP-945), VS-4718 (PND-1186, SR-2516), TAE226, and GSK2256098, or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is selected from VS-4718 (PND-1186, SR-2516), TAE226, and GSK2256098, or a pharmaceutically acceptable salt thereof; or the FAK inhibitor is VS-4718. The combination drug combination of any one of claims 1-2, wherein the pan-RAS(ON) inhibitor is selected from any of the compounds disclosed in WO2022060836A, WO2024067857A1, CN117534684A, CN117534687A, CN117534685A, WO2024060966A1, WO2024104364A1, WO2024169914A1, WO2024017859A1, WO2023025832A1, WO2024008834A1, WO2023232776A1, WO2025119392A1, WO2025087431A1, WO2024208934A1, WO2025045233A1, WO2024249299A2, WO2024222864A1, WO2024211712A, WO2024211663A, and WO2024153208A9. The combination of claim 1-2, wherein the pan-RAS(ON) inhibitor is selected from a compound of Formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof: wherein, selected from the group consisting of n is selected from 0, 1, 2, and 3; A is selected from C3-C 12 Cycloalkylene, 4-10 membered heterocyclic alkylene, C6-C 10 arylene and 5-12-membered heteroarylene, the C3-C 12 Cycloalkylene, 4-10 membered heterocyclic alkylene, C6-C 10 arylene and 5-12 heteroarylene are optionally enclosed by one or more R a replace; R 1 selected from C(O)R 11 , C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl are optionally substituted with 1 or more R 1a ; R 2 R 3 R 4 R 7 R 8 and R 9 Independently selected from hydrogen, halogen, hydroxyl, mercapto, amino, cyano, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkyl groups and C3-C7 cycloalkyl groups; or R 4 and the atom to which they are attached together form a 4-10 membered heterocyclyl ring, which is optionally substituted with 1 or more R 7 and the atom to which they are attached together form a 4-10 membered heterocyclyl ring, which is optionally substituted with 1 or more R b substituents; R 5 selected from the group consisting of C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl and 5-10 membered heteroaryl, said C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl and 5-10 membered heteroaryl are optionally substituted with 1 or more R 5a ; R 6 selected from hydrogen, halogen, hydroxyl, thiol, amino, cyano, Ci-C4alkyl, Ci-C4haloalkyl, and Ci-C4alkoxy; R 11 selected from amino, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said amino, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl are optionally substituted with 1 or more R 11a ; Each R a and R 10 Independently selected from halogens, amino groups, hydroxyl groups, mercapto groups, cyano groups, and C1-C4 alkyl groups; each R 1a and R 11a is independently selected from the group consisting of halogen, amino, hydroxyl, thiol, cyano, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said amino, hydroxyl, thiol, C1-C 10 alkyl, C3-C 12 cycloalkyl, 4-10 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl is optionally substituted with 1 or more R 1aa ; Each R 1aa Independently selected from halogen, amino, hydroxyl, mercapto, cyano, C1-C7 alkyl, C1-C7 haloalkyl and C1-C7 alkoxy; each R is independently selected from the group consisting of 5a is independently selected from the group consisting of C1-C 10 alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkoxy, C3-C 12 cycloalkyl, 4-14 membered heterocyclyl, C6-C 10 aryl and 5-10 membered heteroaryl, said C1-C 10 alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, C1-C 10 alkoxy, C3-C 12 cycloalkyl, 4-14 membered heterocyclyl, C6-C 10 aryl and 5-10 membered heteroaryl are optionally substituted with 1 or more R c substituents; Each R c Independently selected from halogen, amino, hydroxyl, mercapto, cyano, C1-C7 alkyl, C1-C7 alkoxy, C3-C 10 Cycloalkyl and 4-12 membered heterocyclic groups, wherein the amino, hydroxyl, mercapto, C1-C7 alkyl, C1-C7 alkoxy, C3-C 10 Cycloalkyl groups and 4-12 membered heterocyclic groups are optionally surrounded by one or more R groups. h replace; R 5b and R 5b’ are independently selected from the group consisting of C1-C7alkyl, C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl, said C1-C7alkyl, C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, C6-C 10 aryl, and 5-10 membered heteroaryl are optionally substituted with 1 or more R d ; or R 5b and the atom to which they are attached together form a 4-12 membered heterocyclyl group optionally substituted with 1 or more R 5b’ and the atom to which they are attached together form a 4-12 membered heterocyclyl group optionally substituted with 1 or more R e substituents; each R is independently selected from the group consisting of halogen, hydroxyl, amino, C1-C7alkyl, C1-C4alkoxy, C3-C6cycloalkyl, 4-10 membered heterocyclyl, said amino, C1-C7alkyl, C1-C4alkoxy, C3-C6cycloalkyl, 4-10 membered heterocyclyl optionally substituted with 1 or more R d and R e is independently selected from the group consisting of halogen, hydroxyl, amino, C1-C7alkyl, C1-C4alkoxy, C3-C6cycloalkyl, 4-10 membered heterocyclyl, said amino, C1-C7alkyl, C1-C4alkoxy, C3-C6cycloalkyl, 4-10 membered heterocyclyl optionally substituted with 1 or more R f substituents; Each R f Independently selected from C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, and 4-10 membered heterocyclic groups, wherein the C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, and 4-10 membered heterocyclic groups are optionally surrounded by one or more R... g replace; Each R b R g and R h Independently selected from halogens, hydroxyl groups, mercapto groups, amino groups, =O, C1-C4 alkyl groups, C1-C4 hydroxyalkyl groups, C1-C4 haloalkyl groups, (C1-C4 alkylene)O C1-C4 alkyl groups, and C1-C4 alkoxy groups; one or more hydrogen atoms of the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, are optionally deuterium atoms. The combination of claim 4, wherein the compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof does not include (RMC-6236). The combination of claim 4, wherein the compound of formula (I) is selected from the group consisting of or selected from the group consisting of and The combination of any one of claims 4 to 6, wherein A in formula (I) is selected from C6-C 10 arylene and 5-12 membered heteroarylene, said C6-C 10 arylene and 5-12 membered heteroarylene is optionally substituted with 1 or more R a substituents; or A is selected from 5-12 membered heteroarylene, said 5-12 membered heteroarylene is optionally substituted with 1 or more R a substituents; or A is selected from 5-6 membered heteroarylene, said 5-6 membered heteroarylene is optionally substituted with 1 or more R a substituents; or A is selected from a thiazole ring, said thiazole ring is optionally substituted with 1 or more R a substituents; or A is selected from The optionally substituted by 1 or more R a substituted; or A is or A is wherein * represents the attachment point to the indole ring. The combination of drugs according to any one of claims 4-7, wherein R in formula (I) 1 Selected from C(O)R 11 C1-C 10 Alkyl, C3-C 12 Cycloalkyl and 4-10 membered heterocyclic groups, the C1-C 10 Alkyl, C3-C 12 Cycloalkyl groups and 4-10-membered heterocyclic groups are optionally surrounded by one or more R groups. 1a Replace; or R 1 Selected from C(O)R 11 C1-C 10 Alkyl, C4-C 12 Cycloalkyl and 4-10 membered heterocyclic groups, the C1-C 10 Alkyl, C4-C 12 Cycloalkyl groups and 4-10-membered heterocyclic groups are optionally surrounded by one or more R groups. 1a Replace; or R 1 Selected from C(O)R 11 C1-C5 alkyl, C3-C5 cycloalkyl, and 4-6 membered heterocyclic groups, wherein the C1-C5 alkyl, C3-C5 cycloalkyl, and 4-6 membered heterocyclic groups are optionally surrounded by one or more R groups. 1a Replace; or R 1 Selected from C(O)R 11 C1-C5 alkyl, C4-C5 cycloalkyl, and 4-6 membered heterocyclic groups, wherein the C1-C5 alkyl, C4-C5 cycloalkyl, and 4-6 membered heterocyclic groups are optionally surrounded by one or more R groups. 1a Replace; or R 1 Selected from C(O)R 11 methyl, pentyl, cyclopropyl, and tetrahydropyrrole, wherein the methyl, pentyl, cyclopropyl, and tetrahydropyrrole groups are optionally surrounded by one or more R groups. 1a Replace; or R 1 Selected from C(O)R 11 methyl, pentyl, and tetrahydropyrrolyl, wherein the methyl, pentyl, and tetrahydropyrrolyl groups are optionally surrounded by one or more R groups. 1a replace. The combination of any one of claims 4-8, wherein each R 1a is independently selected from the group consisting of halogen, C1-C4 alkyl, and C3-C4 cycloalkyl, said C1-C4 alkyl and C3-C4 cycloalkyl optionally substituted with one or more R 10 ; or each R 12 is independently selected from the group consisting of halogen, C1-C4 alkyl, and C3-C4 cycloalkyl, said C1-C4 alkyl and C3-C4 cycloalkyl optionally substituted with one or more R 10 ; or each R 12 is independently selected from the group consisting of halogen, C1-C4 alkyl, and C3-C4 cycloalkyl, said C1-C4 alkyl and C3-C4 cycloalkyl optionally substituted with one or more R 1aa ; or each R 1a is independently selected from the group consisting of halogen, C1-C4 alkyl, and C3-C4 cycloalkyl, said C1-C4 alkyl and C3-C4 cycloalkyl optionally substituted with one or more R 1aa ; or each R 1a is independently selected from the group consisting of halogen, C1-C4 alkyl, and C3-C4 cycloalkyl, said C1-C4 alkyl and C3-C4 cycloalkyl optionally substituted with one or more R 1aa . The combination of any one of claims 4 to 9, wherein R 11 is selected from 4-10 membered heterocyclyl optionally substituted with 1 or more R 11a ; or R 11 is selected from 4-6 membered heterocyclyl optionally substituted with 1 or more R 11a ; or R 11 is selected from tetrahydropyrrolyl optionally substituted with 1 or more R 11a . The combination of any one of claims 4-10, wherein each R 11a is independently selected from the group consisting of halogen, amino, hydroxyl, thiol, cyano, and C1-C 10 alkyl, said amino, hydroxyl, thiol, and C1-C 10 alkyl are optionally substituted with 1 or more R 1aa ; or each R 11a is independently selected from the group consisting of C1-C 10 alkyl, said C1-C 10 alkyl are optionally substituted with 1 or more R 1aa ; or each R 11a is independently selected from the group consisting of C1-C4 alkyl, said C1-C4 alkyl are optionally substituted with 1 or more R 1aa ; or each R 11a is independently selected from the group consisting of methyl, said methyl are optionally substituted with 1 or more R 1aa . The combination of any one of claims 4-11, wherein each R in Formula (I) 1aa is independently selected from the group consisting of halogen, Ci-C7alkyl, Ci-C7haloalkyl, and Ci-C7alkoxy; or each R 1aa is independently selected from the group consisting of halogen and Ci-C4alkyl; or each R 1aa is independently selected from the group consisting of methyl, chloro, and fluoro. The combination of any one of claims 4 to 12, wherein R 2 , R 3 are independently selected from the group consisting of hydrogen, halogen, hydroxyl, cyano and C1-C 10 alkyl; or R 2 , R 3 are independently selected from the group consisting of C1-C4 alkyl, such as methyl; or R 2 , R 3 are both methyl. The combination of any one of claims 4 to 13, wherein R in formula (I) is selected from the group consisting of hydrogen, halogen, hydroxy, cyano, C1-C4 alkyl and C1-C4 haloalkyl; or R 4 is selected from the group consisting of hydrogen, halogen, hydroxy, cyano, C1-C 10 alkyl and C1-C 10 haloalkyl; or R 4 is selected from the group consisting of C1-C4 alkyl and C1-C4 haloalkyl; or R 4 is selected from the group consisting of ethyl and halo-substituted ethyl; or R 4 is selected from the group consisting of ethyl and fluoro-substituted ethyl, such as trifluoroethyl. The combination of any one of claims 4-14, wherein R 5 is selected from C6-C 10 aryl and 5-10 membered heteroaryl, said C6-C 10 aryl and 5-10 membered heteroaryl are optionally substituted with 1 or more R 5a ; or R 5 is selected from 5-6 membered heteroaryl, said 5-6 membered heteroaryl is optionally substituted with 1 or more R 5a ; or R5 is selected from pyridyl, said N in the pyridyl is optionally oxidized ( + N-O - ), said pyridyl is optionally substituted with 1 or more R 5a ; or R 5 is The optionally substituted by 1 or more R 5a ; or R 5 is wherein N can be oxidized to form The optionally substituted by 1 or more R 5a substituted. The combination of any one of claims 4-15, wherein each R in formula (I) 5a is independently selected from C1-C 10 alkyl, C2-C 10 alkenyl, C1-C 10 alkoxy and 4-14 membered heterocyclyl, said C1-C 10 alkyl, C2-C 10 alkenyl, C1-C 10 alkoxy and 4-14 membered heterocyclyl are optionally substituted with 1 or more R c each R 5a is independently selected from C1-C4alkyl, C2-C4alkynyl, C1-C4alkoxy, and 4-10 membered heterocyclyl, optionally substituted with 1 or more R c each R 5a is independently selected from morpholino, piperazino, piperidino, propynyl, methoxy, ethyl, ethoxy, isopropyl, said morpholinyl, piperazinyl, piperidinyl, propynyl, methoxy, ethyl, ethoxy, isopropyl, and optionally substituted by 1 or more R c ; or each R 5a is independently selected from isopropyl, morpholinyl, The combination of any one of claims 4-16, wherein each R c is independently selected from the group consisting of amino, C1-C7 alkyl, C1-C7 alkoxy, C3-C 10 cycloalkyl, and 4-12 membered heterocyclyl, said amino, C1-C7 alkyl, C1-C7 alkoxy, C3-C 10 cycloalkyl, and 4-12 membered heterocyclyl is optionally substituted with one or more R h ; or each R c is independently selected from the group consisting of amino, C1-C4 alkyl, C1-C4 alkoxy, C3-C8 cycloalkyl, and 4-8 membered heterocyclyl, said amino, C1-C4 alkyl, C1-C4 alkoxy, C3-C8 cycloalkyl, and 4-8 membered heterocyclyl is optionally substituted with one or more R h ; or each R c is independently selected from the group consisting of amino, methyl, ethyl, methoxy, cyclopropyl, morpholinyl, azetidinyl, oxetanyl, piperidinyl, said amino, methyl, ethyl, methoxy, cyclopropyl, morpholinyl, azetidinyl, oxetanyl, and piperidinyl is optionally substituted with 1 or more R h substituents. The combination of any one of claims 4-17, wherein each R in Formula (I) h is independently selected from the group consisting of halogen, hydroxyl, =0, C1-C4alkyl, C1-C4alkyleneOC1-C4alkyl, and C1-C4hydroxyalkyl; or each R h is independently selected from the group consisting of fluorine, hydroxyl, =0, hydroxymethyl, methoxyethyl, and methyl. The combination of any one of claims 4-18, wherein R 5b and the atom to which they are attached together form a 4-6 membered heterocyclyl optionally substituted with 1 or more R 5b’ and the atom to which they are attached together form a 4-6 membered heterocyclyl optionally substituted with 1 or more R e and the atom to which they are attached together form a 4-6 membered heterocyclyl optionally substituted with 1 or more R 5b and the atom to which they are attached together form a 4-6 membered heterocyclyl optionally substituted with 1 or more R 5b’ and the atom to which they are attached together form a 4-6 membered heterocyclyl optionally substituted with 1 or more R e and the atom to which they are attached together form a 4-6 membered heterocyclyl optionally substituted with 1 or more R The combination of any one of claims 4-19, wherein the combination of the compound of formula (I) is selected from the group consisting of The optionally substituted by 1 or more R e substituted. The combination of any one of claims 4-20, wherein each R e is independently selected from the group consisting of halogen, hydroxyl, amino, C1-C7 alkyl, and 4-10 membered heterocyclyl, said hydroxyl, amino, C1-C7 alkyl, and 4-10 membered heterocyclyl optionally substituted with one or more R f ; or each R e is independently selected from the group consisting of C1-C4 alkyl and 4-6 membered heterocyclyl, said C1-C4 alkyl and 4-6 membered heterocyclyl optionally substituted with one or more R f ; or each R e is independently selected from the group consisting of ethyl and morpholinyl, said ethyl and morpholinyl optionally substituted with one or more R f . The combination of any one of claims 4-21, wherein each R f is independently selected from the group consisting of C1-C4alkoxy; or R f is methoxy. The combination of any one of claims 4 to 22, wherein R in formula (I) is C1-C4 alkyl substituted cyclopropyl, and R is selected from isopropyl, 1 5a ​​ or R 1 is C1-C4alkyl substituted cyclopropyl, and R 5a is selected from isopropyl, The combination of any one of claims 4 to 23, wherein R in formula (I) is 6 selected from the group consisting of hydrogen, halogen, hydroxyl, thiol, C1-C4 alkyl and C1-C4 alkoxy; or R 6 selected from the group consisting of hydrogen and C1-C4 alkoxy; or R 6 is hydrogen and ethoxy. The combination of any one of claims 4 to 24, wherein R 7 is selected from hydrogen, halogen, hydroxyl, and cyano; or R 7 is hydrogen. The combination of any one of claims 4-25, wherein R 4 and the atom to which they are attached together form a 6-7 membered heterocyclic ring, which is optionally substituted with 1 or more R 7 and the atom to which they are attached together form a 6-7 membered heterocyclic ring, which is optionally substituted with 1 or more R b substituents. The combination of any one of claims 4-26, wherein each R b is independently selected from the group consisting of halogen, amino, hydroxyl, thiol, and cyano; or each R b is independently selected from the group consisting of halogen, such as fluorine. The combination of any one of claims 4 to 27, wherein R in formula (I) is selected from the group consisting of hydrogen, halogen, hydroxy, cyano, Ci-C4alkyl and C3-C7cycloalkyl; or R 8 is selected from the group consisting of hydrogen, halogen, hydroxy, cyano, Ci-C4alkyl and C3-C7cycloalkyl; or R 10 is selected from the group consisting of hydrogen, halogen, hydroxy, cyano, Ci-C4alkyl and C3-C7cycloalkyl; or R 8 is selected from the group consisting of hydrogen, halogen, hydroxy, cyano, Ci-C4alkyl and C3-C7cycloalkyl; or R 8 is hydrogen. The combination of any one of claims 4 to 28, wherein R 9 is selected from hydrogen, halogen and hydroxyl; or R 9 is selected from hydrogen and halogen; or R 9 is selected from hydrogen and fluorine. The combination drug combination according to claim 4, wherein the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from a compound of formula (I-1) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5a , R 6 , R 7 , R 8 , R 9 , R 10 and n are as defined in claim 4. The combination drug combination according to claim 4, wherein the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from a compound of formula (I-2) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1 , R 2 , R 3 , R 4 , R 5a , R 6 , R 7 , R 8 and R 9 are as defined in claim 4. The combination drug combination of claim 4, wherein the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, (RMC-6236), (RMC-7977), (hereinafter referred to as Compound PA-1), (hereinafter referred to as Compound PA-2), (hereinafter referred to as Compound PA-3), (hereinafter referred to as Compound PA-4), (hereinafter referred to as Compound PA-5), (Hereinafter referred to as Compound PA-6) The combination drug combination of claim 4, wherein the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from RMC-6236, RMC-7977, Compound PA-1, Compound PA-2, Compound PA-3, Compound PA-4, Compound PA-5, and Compound PA-6, or a pharmaceutically acceptable salt thereof; or the compound of Formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is Compound PA-1. A method of treating a neoplastic disease in a subject in need thereof, the method of treatment comprising administering to the subject a pan-RAS(ON) inhibitor and a FAK inhibitor, thereby treating the subject. The method of treatment of claim 34, wherein the FAK inhibitor is selected from the FAK inhibitors of claim 2. The method of treatment of claim 34 or 35, wherein the pan-RAS(ON) inhibitor is selected from the pan-RAS(ON) inhibitors of any one of claims 3-33. A pharmaceutical composition comprising a pan-RAS(ON) inhibitor and a FAK inhibitor, and a pharmaceutically acceptable excipient. The pharmaceutical composition of claim 37, wherein the FAK inhibitor is selected from the FAK inhibitors of claim 2. The pharmaceutical composition of claim 37 or 38, wherein the pan-RAS(ON) inhibitor is selected from the pan-RAS(ON) inhibitors of any one of claims 3-33. A combination product comprising a first pharmaceutical composition comprising at least one pan-RAS(ON) inhibitor and a pharmaceutically acceptable excipient, and a second pharmaceutical composition comprising at least one FAK inhibitor and a pharmaceutically acceptable excipient. The combination product of claim 40, wherein the FAK inhibitor is selected from the FAK inhibitors of claim 2. The combination product of claim 40 or 41, wherein the pan-RAS(ON) inhibitor is selected from the pan-RAS(ON) inhibitors of any one of claims 3-33. A method of treating a neoplastic disease, the method comprising administering to an individual in need of such treatment a therapeutically effective amount of a combination pharmaceutical composition of any one of claims 1-33, a pharmaceutical composition of any one of claims 37-39, or a combination product of any one of claims 40-42. The method of treating a neoplastic disease of any one of claims 34-36, 43, wherein the neoplastic disease comprises, but is not limited to, lung cancer, colorectal cancer, pancreatic cancer, endometrial cancer, myeloma, breast cancer, ovarian cancer, gastric cancer, esophageal cancer, prostate cancer, cervical cancer, cholangiocarcinoma, or uterine carcinosarcoma.

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