Tricyclic compounds, pharmaceutical compositions thereof and uses thereof

Abstract

The present application belongs to the field of pharmaceutical chemistry, and relates to tri-cyclic compounds, pharmaceutical compositions thereof and uses thereof. In particular, the present application relates to tri-cyclic compounds as shown in Formula I, pharmaceutical compositions comprising the same, and uses thereof as SOS1 inhibitors in the medical field. The tri-cyclic compounds in the present application exhibit excellent biological activity and drugability, and have great prospects for drug development.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a class of tricyclic compounds, pharmaceutical compositions containing such compounds, and their applications in the pharmaceutical field. Background Technology

[0002] Since the discovery in late 1982 that the RAS family of GTPases (including KRAS, NRAS, and HRAS) are associated with cancer, they have been found to account for 20%–30% of human cancers. RAS proteins act as molecular switches, cycling between an active GTP-binding state and an inactive GDP-binding state. Activated by guanine nucleotide exchange factor (GEF), the GTP-binding RAS interacts with numerous effectors. The return to the inactive state is driven by GTPase activators (GAPs), which downregulate active RAS by accelerating the weak intrinsic GTPase activity by up to five orders of magnitude.

[0003] Whether mutant RAS proteins require GEF activity for full activation remains to be fully investigated and may vary depending on the specific mutation. The most studied RAS sevenless son (SOS) protein is known to have two human isotypes, SOS1 and SOS2. Attempts were made to mimic the RAS-SOS interaction of a hydrocarbon-bound peptide with nanomolar affinity by inhibiting peptide recognition of the orthogonal SOS helix, but only with low cellular activity. Fragment-based screening, with a well-designed and high-throughput screening method, led to the identification of small-molecule addressing KRAS-SOS1 interactions, yielding proteins with moderate micromolar affinity.

[0004] The SOS1 protein consists of 1333 amino acids (150 kDa). SOS1 is a multidomain protein with two tandem N-terminal histone domains (HD), followed by a Dbl homology domain (DH), a Pleckstrin homology domain (PH), a helical linker (HL), a RAS exchange motif (REM), a CDC25 homology domain, and a C-terminal proline-rich domain (PR). SOS1 has two binding sites for RAS family proteins: a catalytic site that binds GDP-binding RAS family proteins to promote guanine nucleotide exchange; and an allosteric site that binds GTP-binding RAS family proteins, leading to a further enhancement of SOS1's catalytic GEF function. Public data indicate that SOS1 is crucially involved in mutant KRAS activation and oncogenic signaling in cancer (Jeng et al., Nat. Commun., 2012, 3:1168). Depletion of SOS1 levels reduced the proliferation and survival of tumor cells carrying KRAS mutations, but no effect was observed in KRAS wild-type cell lines. The effects of SOS1 loss could not be compensated for by introducing SOS1 with catalytic site mutations, demonstrating the important role of SOS1GEF activity in KRAS-mutant cancer cells.

[0005] SOS1 is crucially involved in the activation of RAS family protein signaling in cancer through mechanisms other than mutations in RAS family proteins. SOS1 interacts with the adaptor protein Grb2, and the resulting SOS1 / Grb2 complex binds to activated / phosphorylated receptor tyrosine kinases (e.g., EGFR, ErbB2, ErbB3, ErbB4, PDGFR-A / B, FGFR1 / 2 / 3, IGF1R, INSR, ALK, ROS, TrkA, TrkB, TrkC, RET, c-MET, VEGFR1 / 2 / 3, AXL) (Pierre et al., Biochem. Pharmacol., 2011, 82(9):1049-56). SOS1 is also recruited to other phosphorylated cell surface receptors, such as the T-cell receptor (TCR), B-cell receptor (BCR), and monocyte colony-stimulating factor receptor (Salojin et al., J. Biol. Chem. 2000, 275(8):5966-75). This localization of SOS1 to the plasma membrane proximal to RAS family proteins enables SOS1 to promote the activation of RAS family proteins. SOS1 activation of RAS family proteins can also be mediated through the interaction of SOS1 / Grb2 with BCR-ABL oncoprotein, which is common in chronic myeloid leukemia.

[0006] SOS1 is also a GEF that activates the GTPase RAC1 (Ras-associated C3 botulinum toxin substrate 1) (Innocenti et al., J. Cell Biol., 2002, 156(1):125-36). Like RAS family proteins, RAC1 is involved in the pathogenesis of various human cancers and other diseases (Bid et al., Mol. Cancer Ther. 2013, 12(10):1925-34).

[0007] In this paper, we describe novel SOS1 inhibitor compounds that bind to the SOS1 catalytic site and simultaneously prevent interaction with and activation of RAS family proteins. This results in a significant inhibition of the interaction between SOS1 and RAS family proteins, particularly KRAS (which has low nanomolar IC50 activity), and consequently a significant reduction in ERK phosphorylation in KRAS mutant cancer cell lines.

[0008] The selective SOS1 inhibitor compounds described herein are expected to provide pharmacological benefits to patients with cancers associated with dependence on RAS family protein signaling. Such cancers expected to be targeted by SOS1 inhibitor compounds include those exhibiting alterations (mutations, gene amplification, overexpression) of components (proteins, genes) in the RAS family protein pathway, such as KRAS, NRAS, HRAS, receptor tyrosine kinases (e.g., EGFR, ErbB2, ErbB3, ErbB4, PDGFR-A / B, FGFR1 / 2 / 3, IGF1R, INSR, ALK, ROS, TrkA, TrkB, TrkC, RET, c-MET, VEGFR1 / 2 / 3, AXL), GAP (e.g., NF1), and SOS1. Furthermore, given the role of SOS1 in RAC1 activation, cancers exhibiting dependence on RAC1 are expected to be targeted by SOS1 inhibitor compounds. In addition, SOS1 inhibitor compounds are expected to provide pharmacological benefits in other diseases associated with dysregulation of RAS family protein pathways, such as neurofibromatosis, Noonan syndrome (NS), cardiofacial skin syndrome (CFC), and hereditary gingival fibromatosis type 1.

[0009] In addition to their inhibitory effects and potency, the compounds disclosed herein exhibit good solubility, excellent DMPK properties, and good selectivity for kinases in the human kinase community. Summary of the Invention

[0010] The problem the invention aims to solve

[0011] The present invention aims to provide a class of novel tricyclic compounds for use as SOS1 inhibitors, which exhibit good inhibitory activity against tumor cells, have good drug-like properties, and have broad prospects for drug development.

[0012] Solution for solving the problem

[0013] In a first aspect, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, wherein...

[0014]

[0015] A is selected from C6-C 10 Aryl, 5- to 6-membered monocyclic heteroaryl and 9- to 10-membered bicyclic heteroaryl, wherein each of the aryl, monocyclic heteroaryl and bicyclic heteroaryl is optionally substituted by up to 5 R6s;

[0016] X and Y are each independently selected from CR6 and N;

[0017] Q1 and Q2 are each independently selected from -O-, -C(R9)2- and -NR9-;

[0018] L1 and L2 are each independently selected from -(CH2). m -or-(CH2) m -O-(CH2) p -O-(CH2) n - where each m, n, and p is an independent integer from 0 to 8;

[0019] R1 and R2 are each independently selected from hydrogen and C1-C8 alkyl groups; or R1 and R2 together with the carbon atoms they are attached to form C3-C6 cycloalkyl groups;

[0020] R3 is selected from hydrogen, halogen, cyano, hydroxyl, amino, -NH(R6), -C(=O)-NH(R6), C1-C6 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl, 3 to 8-membered heterocyclic alkyl, C1-C3 alkoxy and C1-C6 haloalkyl, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic alkyl, alkoxy and haloalkyl is optionally substituted by at least one R6;

[0021] R4 is selected from hydrogen, halogen, cyano, hydroxyl, amino, -NH(R6), -C(=O)-NH(R6), C1-C6 alkyl, C3-C6 cycloalkyl, 3 to 8-membered heterocyclic alkyl, C1-C3 alkoxy and C1-C6 haloalkyl, wherein each of the alkyl, cycloalkyl, heterocyclic alkyl, alkoxy and haloalkyl is optionally substituted by at least one R6;

[0022] R5 is selected from hydrogen, halogen, cyano, hydroxyl, amino, -N(R6)(R7), -C(=O)-N(R6)(R7), -C(=O)-R7, -C(=O)-OR7, C1-C6 alkyl, C3-C6 cycloalkyl, 3 to 8-membered heterocyclic alkyl, C1-C3 alkoxy and C1-C6 haloalkyl, wherein each of the alkyl, cycloalkyl, heterocyclic alkyl, alkoxy and haloalkyl is optionally substituted by at least one R6;

[0023] If present, each R6 and R7 is independently selected from hydrogen, halogen, cyano, hydroxyl, amino, carbamoyl, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C8 cycloalkyl, 3- to 14-membered heterocycloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, C6-C 10 The alkyl, 5- to 6-membered monocyclic heteroaryl, and 9- to 10-membered bicyclic heteroaryl groups are each optionally substituted with at least one R8 atom; or R6 and R7 together with the nitrogen atom to which they are attached form a 5- to 6-membered heterocyclic alkyl group, wherein the heterocyclic alkyl group is optionally substituted with at least one R8 atom.

[0024] If present, each R8 is independently selected from hydrogen, chlorine, fluorine, cyano, hydroxyl, amino, isopropyl, cyclopropyl, methyl, difluoromethyl, trifluoromethyl, methoxy, trifluoromethoxy, ethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy, and phenyl.

[0025] If present, each R9 is independently selected from hydrogen, halogen, cyano, hydroxyl, amino, -N(R6)(R7), -C(=O)-N(R6)(R7), -C(=O)-R7, -C(=O)-OR7, C1-C6 alkyl, C3-C6 cycloalkyl, 3 to 8-membered heterocyclic alkyl, C1-C3 alkoxy, and C1-C6 haloalkyl, wherein each of the alkyl, cycloalkyl, heterocyclic alkyl, alkoxy, and haloalkyl is optionally substituted by at least one R6.

[0026] Preferably, the compound shown in Formula I is the compound shown in Formula I-1, wherein

[0027]

[0028] q is any integer from 0 to 4; X, Y, R1, R2, R3, R4, R5 and R6 are as defined in Equation 1.

[0029] More preferably, the compound represented by Formula I or Formula I-1 is the compound represented by Formula I-1-1 or Formula I-1-2, wherein

[0030]

[0031] q is any integer from 0 to 4; X, Y, R1, R2, R3, R4, R6 and R7 are as defined in Equation 1.

[0032] More preferably, in the compound shown in Formula I-1-1,

[0033] The radical group is selected from any one of the following radical groups:

[0034]

[0035] Preferably, the compound shown in Formula I is a compound shown in Formula I-2, wherein

[0036]

[0037] q is any integer from 0 to 4; X, Y, R1, R2, R3, R4, R5, and R6 are as defined in Formula 1. More preferably, the compound shown in Formula I or Formula I-2 is the compound shown in Formula I-2-1, wherein...

[0038]

[0039] q is any integer from 0 to 4; X, Y, R1, R2, R3, R4 and R6 are as defined in Equation 1.

[0040] Preferably, the compound shown in Formula I is a compound shown in Formulas I-3, wherein

[0041]

[0042] q is any integer from 0 to 4; X, Y, R1, R2, R3, R4, R5, R6, and R9 are as defined in Formula 1. More preferably, the compound shown in Formula I or Formula I-3 is the compound shown in Formula I-3-1, wherein...

[0043]

[0044] q is any integer from 0 to 4; X, Y, R1, R2, R3, R4, R6 and R9 are as defined in Equation 1.

[0045] More preferably, in the compound shown in Formula I-3-1,

[0046] R9 is selected from any one of the following groups:

[0047]

[0048] In a second aspect, the present invention provides specific compounds as shown in Formula I, Formula I-1, Formula I-1-1, Formula I-2, Formula I-2-1, Formula I-3, or Formula I-3-1, selected from:

[0049] N-(1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)-2-methyl-7,8-dihydro-[1,4]dioxano[2,3-g]quinazolin-4-amine;

[0050] N-(1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)-6-benzyl-2-methyl-7,8-dihydro-6H-[1,4]oxazino[3,2-g]quinazolin-4-amine;

[0051] 1-(4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2-methyl-7,8-dihydro-6H-[1,4]oxazinofo[3,2-g]quinazolin-6-yl)-1-ethyl ketone;

[0052] (R)-(4-((1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)amino)-2-methyl-7,8-dihydro-6H-[1,4]oxazino[3,2-g]quinazolin-6-yl)(1-methylpiperidin-4-yl)methyl ketone;

[0053] (R)-(4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-7,8-dihydro-6H-[1,4]oxazino[3,2-g]quinazolin-6-yl)(1-methylpiperidin-4-yl)methyl ketone;

[0054] 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-7,8-dihydro-[1,4]dioxin[2,3-g]quinazolin-7-carboxylic acid ethyl ester; and

[0055] 4-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-7,8-dihydro-[1,4]dioxin[2,3-g]quinazolin-8-carboxylic acid ethyl ester.

[0056] Thirdly, the present invention provides a pharmaceutical composition comprising a compound of formula I, formula I-1, formula I-1-1, formula I-1-2, formula I-2, formula I-2-1, formula I-3 or formula I-3-1, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, and at least one pharmaceutically acceptable excipient.

[0057] Fifthly, the present invention provides compounds of formula I, formula I-1, formula I-1-1, formula I-1-2, formula I-2, formula I-2-1, formula I-3 or formula I-3-1, or pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, metabolites or prodrugs thereof, or pharmaceutical compositions comprising thereof, for use as SOS1 inhibitors or for prevention and / or treatment of diseases or conditions caused by SOS1 overexpression.

[0058] In a sixth aspect, the present invention provides the use of compounds of formula I, formula I-1, formula I-1-1, formula I-1-2, formula I-2, formula I-2-1, formula I-3 or formula I-3-1, or pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, metabolites or prodrugs thereof, or pharmaceutical compositions comprising thereof, in the preparation of medicaments for the prevention and / or treatment of diseases or conditions caused by SOS1 overexpression.

[0059] In a seventh aspect, the present invention provides a method for preventing and / or treating diseases or conditions caused by SOS1 overexpression, comprising administering a preventive and / or therapeutically effective amount of a compound of formula I, I-1, I-1-1, I-1-2, I-2, I-2-1, I-3 or I-3-1, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug or a pharmaceutical composition comprising the same, to an individual in need.

[0060] Preferably, in the above-mentioned pharmaceutical use, the disease or condition caused by SOS1 overexpression is cancer, preferably pancreatic cancer, colorectal cancer, and lung cancer.

[0061] The effects of the invention

[0062] This invention provides a series of novel tricyclic compounds. Enzyme and cell activity assays have demonstrated that these compounds possess excellent cell proliferation inhibitory activity. In in vitro experiments, they exhibit an IC50 inhibitory effect on cell proliferation. 50 With values ​​reaching the nM level, it can be well applied in various tumors. Simultaneously, the compounds of this invention exhibit very good inhibitory effects on KRAS:SOS1 activation, reaching the nM level, making them suitable for preparation as SOS1 inhibitors for the prevention and / or treatment of diseases or conditions related to SOS1 activation, such as cancers (including but not limited to pancreatic cancer, colorectal cancer, and lung cancer). Detailed Implementation

[0063] General terms and definitions

[0064] Unless otherwise stated, the terms used in this invention have the following meanings.

[0065] "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 20 carbon atoms, such as straight-chain and branched groups with 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In this invention, "alkyl" can be a monovalent, divalent, or trivalent group. Non-limiting examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, and their various branched isomers. Non-limiting examples also include, but are not limited to, methylene, methine, ethylene, methine, propylene, methine, butylene, methine, and their various branched isomers. Furthermore, in this invention, "alkyl" may be optionally substituted or unsubstituted.

[0066] "Alkoxy" refers to the "-O-alkyl" group, where "alkyl" is defined as described above.

[0067] "Alkenyl" refers to an unsaturated aliphatic hydrocarbon group, comprising straight-chain and branched groups with 1 to 20 carbon atoms and at least one carbon-carbon double bond, such as straight-chain and branched groups with 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In this invention, "alkenyl" can be a monovalent, divalent, or trivalent group. Non-limiting examples include, but are not limited to, vinyl (-CH=CH2), propen-1-yl (-CH=CH-CH3), propen-2-yl (-C(CH3)=CH2), buten-1-yl (-CH=CH-CH2-CH3), buten-2-yl (-C(C2H5)=CH2), 1-methylpropen-1-yl (-C(CH3)=CH-CH3), and their various branched isomers. Non-limiting examples also include, but are not limited to, 1,1-vinylene (=C=CH2), 1,2-vinylene (-CH=CH-), 1,1-propenylidene (=C=CH-CH3), 1,2-propenylidene (-CH=C(CH3)-), 1,3-propenylidene (-CH=CH-CH2-), and their various branched isomers. Furthermore, in this invention, the "alkenyl" may be optionally substituted or unsubstituted.

[0068] "Alynyl" refers to an unsaturated aliphatic hydrocarbon group comprising 1 to 20 carbon atoms and at least one carbon-carbon triple bond, in both straight-chain and branched groups. For example, it can be a straight-chain or branched group with 1 to 18, 1 to 12, 1 to 8, 1 to 6, or 1 to 4 carbon atoms. In this invention, "alkynyl" can be a monovalent, divalent, or trivalent group. Non-limiting examples include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡C-CH3), and butynyl. Pentyne And its various branched isomers, etc. Non-limiting examples also include, but are not limited to, ethynylene (-C≡C-) and propynylene. Butyrynethin And its various branched isomers. Furthermore, in this invention, the "alkynyl group" may be optionally substituted or unsubstituted.

[0069] "Heteroalkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups of 2 to 20 atoms, for example, straight-chain and branched groups of 2 to 18 atoms, 2 to 12 atoms, 2 to 8 atoms, 2 to 6 atoms, or 2 to 4 atoms, wherein one or more atoms are selected from nitrogen, oxygen, or S(O). m (Where m is 0, 1, or 2) heteroatoms, with the remainder being carbon. In this invention, "heteroalkyl" can be a monovalent, divalent, or trivalent group. Non-limiting examples include, but are not limited to, methoxymethyl (2-oxapropyl), methylthiomethyl (2-thiopropyl), methylaminomethyl (2-azapropyl), and their various branched isomers. Furthermore, in this invention, "heteroalkyl" can be optionally substituted or unsubstituted.

[0070] "Cycloalkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group comprising 3 to 12 ring atoms, for example, 3 to 12, 3 to 10, or 3 to 6 ring atoms (i.e., 3 to 6 membered rings). Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc. In this invention, "cycloalkyl" may be optionally substituted or unsubstituted.

[0071] "Heterocyclic alkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group comprising 3 to 20 ring atoms, for example, 3 to 16, 3 to 12, 3 to 10, or 3 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O). m(where m is 0, 1, or 2) heteroatoms, with the remaining ring atoms being carbon. Preferably, the heterocyclic alkyl group comprises 3 to 12 ring atoms, wherein 1 to 4 ring atoms are heteroatoms; more preferably, it comprises 3 to 10 ring atoms; most preferably, it comprises 5 or 6 ring atoms, wherein 1 to 4, preferably 1 to 3, and more preferably 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocyclic alkyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, and homopiperazinyl. Non-limiting examples of polycyclic heterocyclic alkyl groups include, but are not limited to, spirocyclic or bridged heterocyclic alkyl groups.

[0072] "Halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine, chlorine and bromine being preferred.

[0073] "Halogenated alkyl" or "halogenated alkoxy" means that an alkyl or alkoxy group is replaced by one or more identical or different halogen atoms. Preferred examples of alkyl or alkoxy groups include, but are not limited to, trifluoromethyl, trifluoroethyl, and trifluoromethoxy.

[0074] "Cyano" refers to the "-CN" group.

[0075] "Hydroxy group" refers to the "-OH" group.

[0076] "Amino" refers to the "-NH2" group.

[0077] "Carbamoyl" refers to the "-(C=O)-NH2" group.

[0078] "Aryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic system containing 6-14 ring atoms, wherein at least one ring system is aromatic, and each ring system comprises a ring of 3-7 atoms with one or more bonding sites connected to the rest of the molecule. Examples include, but are not limited to, phenyl, naphthyl, and anthracene. Preferably, the aryl group is a carbocyclic system with 6-10 or 6-7 ring atoms.

[0079] "Heteroaryl" refers to a monocyclic, bicyclic, or tricyclic system containing 5-14 ring atoms, wherein at least one ring system is aromatic, and at least one ring system contains one or more heteroatoms selected from nitrogen, oxygen, and sulfur, wherein each ring system contains a ring of 5-7 atoms and has one or more bonding sites connected to the rest of the molecule. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic compound." Examples include, but are not limited to: furanyl, imidazolyl, 2-pyridyl, 3-pyridyl, thiazolyl, purinyl, and quinolinyl. Preferably, the heteroaryl is a ring system with 5-10 ring atoms.

[0080] "Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility that the event or environment may or may not occur. For example, "optionally alkyl-substituted heterocyclic group" means that the alkyl group may but is not required to be present, and the description includes the possibility that the heterocyclic group is substituted with an alkyl group and the possibility that the heterocyclic group is not substituted with an alkyl group.

[0081] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, and more preferably one to three hydrogen atoms that are independently substituted by the corresponding number of substituents.

[0082] "Pharmaceutically acceptable salt" refers to a salt prepared from the compounds of this invention with a relatively non-toxic acid or base. When the compounds of this invention contain a relatively acidic functional group (e.g., a carboxyl or sulfonic acid group), a base addition salt can be obtained by contacting it with a sufficient amount of base in a pure solution or a suitable inert solvent in its free form. Non-limiting examples of pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, ammonium, calcium, magnesium, organic amine, or similar salts. When the compounds of this invention contain a relatively basic functional group (e.g., an amino or guanidine group), an acid addition salt can be obtained by contacting it with a sufficient amount of acid in a pure solution or a suitable inert solvent in its free form. Non-limiting examples of pharmaceutically acceptable acid addition salts include, but are not limited to, inorganic acid salts (e.g., hydrochlorides, hydrobroms, hydroiodates, nitrates, carbonates, bicarbonates, phosphates, monohydrogen phosphates, dihydrogen phosphates, phosphites, sulfates, hydrogen sulfates, etc.), organic acid salts (e.g., acetates, propionates, isobutyrates, malonates, succinates, octanoates, maleates, fumarates, citrates, tartrates, lactates, mandelates, benzoates, phthalates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, glucuronic acid, etc.), and amino acid salts (e.g., arginine salts). Specific forms of pharmaceutically acceptable salts can also be found in Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66:1-19. Certain compounds of this invention contain both basic and acidic functional groups, and thus can be converted into either a basic addition salt or an acid addition salt. Preferably, the salt is contacted with a base or acid in a conventional manner, and then the parent compound is separated, thereby regenerating the neutral form of the compound. The parent form of the compound differs from its various salt forms in certain physical properties, such as different solubilities in polar solvents. According to embodiments of the invention, the pharmaceutically acceptable salt of the compound as shown in Formula I is preferably an acid addition salt, preferably a hydrochloride, hydrobromide, phosphate, or sulfate, more preferably a hydrochloride.

[0083] "Pharmaceutical composition" means a composition that is pharmaceutically usable and comprises one or more compounds as shown in Formula I or in a pharmaceutically acceptable form thereof (e.g., salt, hydrate, solvate, stereoisomer, tautomer, metabolite, prodrug, etc.), and other components (e.g., pharmaceutically acceptable excipients).

[0084] In this invention, "pharmaceuticalally acceptable excipients" refer to auxiliary materials widely used in the pharmaceutical manufacturing field. The primary purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in nature, and / or has specific functionalities, and also to provide a method for enabling the active ingredient to dissolve at a desired rate or promote the effective absorption of the active ingredient in the body of the administered subject after administration of the drug. Pharmaceutically acceptable excipients can be inert fillers or functional ingredients that provide a certain function to the pharmaceutical composition (e.g., stabilizing the overall pH of the composition or preventing the degradation of the active ingredient in the composition). Non-limiting examples of pharmaceutically acceptable excipients include, but are not limited to, binders, suspending agents, emulsifiers, diluents (or fillers), granulating agents, adhesives, disintegrants, lubricants, anti-adhesion agents, flow aids, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, sweeteners, etc.

[0085] The pharmaceutical compositions of this invention can be prepared using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulation, emulsification, grinding, encapsulation, embedding, and / or lyophilization processes.

[0086] In this invention, the purpose of using the pharmaceutical composition is to promote drug delivery to the organism, facilitating the absorption of the active ingredient and thereby exerting its biological activity. The pharmaceutical compositions of this invention can be administered in any form, including injection (intra-arterial, intravenous, intramuscular, intraperitoneal, subcutaneous), mucosal, oral (oral solid dosage forms, oral liquid dosage forms), rectal, inhalation, implantation, and topical (e.g., ocular) administration. Non-limiting examples of oral solid dosage forms include, but are not limited to, powders, capsules, lozenges, granules, tablets, etc. Non-limiting examples of oral or mucosal liquid dosage forms include, but are not limited to, suspensions, tinctures, elixirs, solutions, etc. Non-limiting examples of topical dosage forms include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Non-limiting examples of parenteral dosage forms include, but are not limited to, solutions for injection, dry powders for injection, suspensions for injection, emulsions for injection, etc. The pharmaceutical compositions of this invention can also be formulated into controlled-release or delayed-release dosage forms (e.g., liposomes or microspheres).

[0087] Preferably, the compounds of the present invention or pharmaceutical compositions comprising them are administered orally or intravenously to the individual in need. Depending on the specific circumstances of the recipient, other administration methods may also be used or even preferred. For example, percutaneous administration would be a very important method of administration for patients who are forgetful or irritable with oral medications. In the present invention, the administration method can be varied or adjusted in any applicable manner to meet the needs of the properties of the drug, the convenience of the patient and healthcare personnel, and other relevant factors.

[0088] The compounds of the present invention, or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, metabolites, or prodrugs, or pharmaceutical compositions comprising them, possess excellent SOS1 enzyme activity and cell proliferation inhibitory activity, and can be used as SOS1 inhibitors for the prevention and / or treatment of diseases or conditions caused by SOS1 overexpression, exhibiting good clinical and pharmaceutical applications. Preferably, non-limiting examples of diseases or conditions caused by SOS1 overexpression are cancers, including but not limited to pancreatic cancer, colorectal cancer, and lung cancer.

[0089] The technical solution of the present invention will be described below with reference to specific embodiments. The following embodiments are provided to further illustrate the present invention, and are not intended to limit the scope of the present invention. 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 invention without departing from the spirit and scope of the invention.

[0090] The compounds of this invention can be prepared by synthetic methods well known to those skilled in the art, including but not limited to the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of this invention. The known starting materials used in this invention can be synthesized by methods known in the art or purchased through conventional commercial means (e.g., from companies such as Shaoyuan Chemical Technology and Beijing Coupling Technology). Unless otherwise specified, all reactions are carried out under an argon or nitrogen atmosphere. Hydrogenation reactions are typically performed under vacuum and purged with hydrogen, repeated three times. The reaction temperature is room temperature, ranging from 20°C to 30°C. Monitoring of the reaction progress can be achieved by synthetic methods well known to those skilled in the art, including but not limited to thin-layer chromatography (TLC). Qingdao Ocean GF254 silica gel plates are used for TLC. The developing solvent system includes, but is not limited to, A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent can be adjusted according to the polarity of the compound.

[0091] The separation and purification of the compounds of this invention can be achieved through synthetic methods well known to those skilled in the art, including but not limited to column chromatography (CC), high-performance liquid chromatography (HPLC), and ultra-high-performance liquid chromatography (UPLC). Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as the carrier. The eluent system includes, but is not limited to, A: dichloromethane and methanol; B: petroleum ether and ethyl acetate. The volume ratio of the solvent can be adjusted according to the polarity of the compound, and a small amount of acidic or basic anti-tailing reagent can also be added for adjustment. HPLC chromatograms are determined using an Agilent 1200DAD HPLC system (column: Sunfire C18, 150×4.6mm, 5μm) or a Waters 2695-2996 HPLC system (column: Gimini C18, 150×4.6mm, 5μm).

[0092] The structural identification of the compounds of this invention can be achieved by methods well known to those skilled in the art, including but not limited to nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR spectra were determined using a Bruker AVANCE-400 or Varian Oxford-300 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), or deuterated methanol (CD3OD) as the solvent, and tetramethylsilane (TMS) as the internal standard. Chemical shifts were expressed in 10⁻⁶ increments. -6 (ppm) were measured. MS spectra were determined using an Agilent SQD (ESI) mass spectrometer (model: 6110) or a Shimadzu SQD (ESI) mass spectrometer (model: 2020).

[0093] Preparation of intermediates

[0094] Preparation of intermediate INT-1

[0095]

[0096] Step 1: Synthesize compound INT-1B

[0097] Compound INT-1A (25 g, 107 mmol) was dissolved in THF (250 mL), and tert-butylsulfinamide (19.5 g, 161 mmol) was added at room temperature, followed by tetraethyl titanate (61 g, 267.5 mmol). After the addition was complete, the reaction mixture was heated to 70 °C and reacted for 4 h. After the reaction was complete as shown by TLC, the reaction mixture was cooled to room temperature and slowly added to ice water. The aqueous phase was extracted with ethyl acetate (3 × 150 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1 (v / v)) to give compound INT-1B (27.7 g, pale yellow solid, 77% yield).

[0098] MS(ESI): m / z 337[M+1] + .

[0099] Step 2: Synthesizing compound INT-1C

[0100] Compound INT-1B (27 g, 80 mmol) was dissolved in a mixed solvent of methanol (200 mL) and water (100 mL), cooled to -20 °C, and then sodium borohydride (6 g, 160 mmol) was added in portions. The reaction was maintained at -20 °C for 1 h, then naturally warmed to room temperature and continued to react at room temperature for 3 h. After the reaction was complete as shown by TLC, methanol was removed by vacuum distillation, and the aqueous phase was extracted with ethyl acetate (3 × 150 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound INT-1C (22 g, pale yellow solid, yield 82%).

[0101] MS(ESI): m / z 339[M+1] + .

[0102] Step 3: Synthesize compound INT-1

[0103] Compound INT-1C (20 g, 59 mmol) was added to 200 mL of 4N HCl-ethyl acetate solution and reacted at room temperature for 2 h. After the reaction was complete, a solid precipitated. The solid was filtered, washed with ethyl acetate, and dried to give INT-1 hydrochloride (12.3 g, pale yellow solid, 89% yield). The product did not require purification and was used directly in subsequent reactions.

[0104] MS(ESI): m / z 235[M+1] + .

[0105] Preparation of target compound

[0106] Example 1: Preparation of Compound 1

[0107]

[0108] Synthesis route:

[0109]

[0110] Preparation method:

[0111] Step 1: Synthesize compound 1B

[0112] Compound 1A (25 g, 104 mmol) was dissolved in a mixed solvent of ethanol (250 mL) and water (50 mL). Iron powder (23 g, 416 mmol) and ammonium chloride (22 g, 416 mmol) were added at room temperature, and the reaction mixture was heated under reflux overnight. After the reaction was complete as shown by TLC, the reaction mixture was filtered, the filtrate was evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1:1 (v / v)) to give compound 1B (16.6 g, yellow solid, 76% yield).

[0113] MS(ESI): m / z 210[M+1] + .

[0114] Step 2: Synthesize compound 1C

[0115] In a 250 mL sealed container, compound 1B (16 g, 76 mmol) was dissolved in 100 mL of 4N hydrochloric acid acetonitrile solution. After sealing, the mixture was heated to 110 °C and reacted for 8 h. TLC showed that after the reaction was complete, the mixture was cooled to room temperature, and a large amount of solid precipitated. The solid was filtered, washed with diethyl ether, and dried to give compound 1C (13.8 g, white solid, yield 83%).

[0116] MS(ESI): m / z 219[M+1] + .

[0117] Step 3: Synthesize compound 1D

[0118] Compound 1C (10 g, 45.6 mmol) was added to phosphorus oxychloride (100 mL), and the mixture was heated to 120 °C and reacted for 6 h. After the reaction was complete (using TLC), phosphorus oxychloride was removed under reduced pressure. The residue was dissolved in ethyl acetate and slowly added to ice water. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (3 × 150 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound 1D (8.7 g, pale yellow solid, 81% yield).

[0119] MS(ESI): m / z 237[M+1] + .

[0120] Step 4: Synthesize compound 1E

[0121] Compound 1D (500 mg, 2.1 mmol) was added to acetonitrile (5 mL), followed by intermediate INT-1 (580 mg, 2.5 mmol) and DIEPA (540 mg, 4.2 mmol). After the addition was complete, the reaction mixture was heated to 80 °C and reacted for 2 h. After the reaction was complete as shown by TLC, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 1E (590 mg, pale yellow solid, yield 65.3%).

[0122] MS(ESI): m / z 435[M+1] + .

[0123] Step 5: Synthesize Compound 1

[0124] Compound 1E (500 mg, 1.1 mmol) was added to methanol (5 mL) at room temperature, followed by 10% palladium on carbon (50 mg). After the addition was complete, a hydrogen balloon was attached to the bottle neck, and the gas was purged three times. The reaction was then carried out at room temperature under a hydrogen atmosphere for 3 h. After the reaction was complete as shown by TLC, the mixture was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was removed by vacuum distillation, and the residue was purified using a preparative silica gel plate (electrolyte: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 1 (300 mg, gray solid, yield 67%).

[0125] MS(ESI): m / z 405[M+1] + .

[0126] 1 H-NMR (400MHz, CDCl3): δ7.21(s,1H),7.12(s,1H),7.07(s,1H),6.91-6.89(m,1H),6.82-6.80(m, 1H),5.62-5.57(m,1H),5.51(brs,1H),4.89-4.31(m,4H),3.87(s,2H),2.57(s,3H),1.65(d,3H).

[0127] Example 2: Preparation of Compound 2

[0128]

[0129] Synthesis route:

[0130]

[0131] Preparation method:

[0132] Step 1: Synthesize compound 2B

[0133] Compound 2A (50 g, 228 mmol) was added to a flask containing concentrated sulfuric acid (425 mL) under ice bath conditions. KNO3 (23.1 g, 228 mmol) was added in portions under ice bath conditions. After the addition was complete, the mixture was stirred under ice bath conditions for 5 min, and then stirred at room temperature for 3 hours. After the reaction was completed as indicated by TLC, the solid was precipitated by adding ice water. The solid was filtered and dried to give compound 2B (49 g, yield 81.6%).

[0134] 1 H-NMR (400MHz, DMSO): δ8.05 (d, J = 10.8 Hz, 1H), 8.60 (d, J = 8.0 Hz, 1H), 14 (s, 1H).

[0135] Step 2: Synthesize compound 2C

[0136] Methanol (480 mL) was added to a three-necked flask, followed by compound 2B (49 g, 185.6 mmol). Under nitrogen protection, SOCl2 (29 g, 24.38 mmol) was added dropwise with stirring in an ice bath. After the addition was complete, the mixture was stirred at room temperature for 10 min and then incubated overnight at 70 °C. TLC showed that the reaction was complete. The mixture was then evaporated to dryness, slurried with petroleum ether / ethyl acetate (5:1 v / v), and dried to give compound 2C (43 g, 83.5% yield).

[0137] 1 H-NMR (400MHz, DMSO): δ7.7 (d, J = 10.0 Hz, 1H), 8.4 (d, J = 7.6 Hz, 1H), 3.9 (s, 3H).

[0138] Step 3: Synthesize compound 2D

[0139] 2-Bromoethanol (10.8 g, 86.4 mmol) was dissolved in THF (100 mL), and triisopropylaminolithium (LDA, 2 mol / L) (54 mL, 108 mmol) was added dropwise at 0 °C. The mixture was stirred at 0 °C for 0.5 h. Then, compound 2C (20 g, 72 mmol) was dissolved in THF and added dropwise to the reaction mixture. The mixture was stirred at room temperature for 3 h. After the reaction was complete as shown by TLC, saturated NH4Cl aqueous solution (500 mL) was added, and the aqueous phase was extracted with ethyl acetate (3 × 150 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound 2D (25 g, yield 90.67%).

[0140] 1 H-NMR (400MHz, CDCl3): δ8.48(s,1H),7.37(s,1H),4.48(t,J=6.3Hz,2H),3.95(s,3H),3.70(t,J=8.0Hz,2H).

[0141] Step 4: Synthesize compound 2E

[0142] Compound 2D (25 g, 65.2 mmol) was dissolved in a mixed solution of ethanol (200 mL) and water (50 mL). Iron powder (11 g, 196.5 mmol) and ammonium chloride (21.25 g, 393 mmol) were added, and the mixture was stirred and refluxed overnight at 80 °C under nitrogen protection. After the reaction was complete as shown by TLC, the mixture was filtered, and the solid was washed several times with ethyl acetate. The filtrate was added to a saturated aqueous solution of NH4Cl (500 mL), and the aqueous phase was separated. The aqueous phase was extracted with ethyl acetate (3 × 150 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound 2E (10.8 g, 61% yield).

[0143] MS(ESI): m / z 272,274[M+1] + .

[0144] Step 5: Synthesize compound 2F

[0145] Compound 2E (5 g, 18.4 mmol) was added to DMF (50 mL) at room temperature, followed by potassium carbonate (5.07 g, 26.8 mmol), and then benzyl bromide (2.82 g, 16.56 mmol) was added dropwise at room temperature. After the addition was complete, the mixture was reacted at room temperature for 3 h. After the reaction was complete, the reaction solution was slowly added to water, and a solid precipitated. The solid was filtered, washed with water, and dried to give the gray product 2F (5.2 g, yield 77.6%).

[0146] MS(ESI): m / z 362,364[M+1] + .

[0147] Step 6: Synthesize compound 2G

[0148] Compound 2F (2.2 g, 6.04 mmol) was dissolved in dioxane (25 mL), followed by diphenyl ketone imine (2 g, 12.08 mmol), Pd₂(dba)₃ (0.24 g, 604 μmol), xantphos (0.35 g, 1.2 mmol), and Cs₂CO₃ (3.9 g, 12.08 mmol). The mixture was stirred and refluxed overnight at 95 °C under nitrogen protection. After the reaction was complete as shown by TLC, the reaction solution was cooled to room temperature and added to a saturated NH₄Cl aqueous solution (100 mL). The aqueous phase was separated and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound 2G (3 g, 99% yield).

[0149] MS(ESI): m / z 463[M+1] + .

[0150] Step 7: Synthesize compound 2H

[0151] Compound 2G (2.8 g, 6.03 mmol) was dissolved in methanol (25 mL), and ethyl hydrochloride solution (5 mL, 8% by mass) was added dropwise at low temperature. The mixture was left to stand overnight at room temperature. After the reaction was complete as shown by TLC, the mixture was evaporated to dryness, slurried with petroleum ether / ethyl acetate (1:1 v / v), and dried to give compound 2H (1.62 g, 90% yield).

[0152] MS(ESI): m / z 299[M+1] + .

[0153] Step 8: Synthesize compound 2I

[0154] Compound 2H (1.16 g, 3.9 mmol) was added to a sealed container containing acetonitrile (10 mL), followed by the addition of ethyl hydrochloride solution (5 mL, 8% by mass). The mixture was stirred overnight at 100 °C. After the reaction was complete as shown by TLC, the mixture was cooled to room temperature, and a solid precipitated. The solid was filtered, washed with cold ethyl acetate, and then dried to give compound 2I (0.7 g, 60% yield).

[0155] MS(ESI): m / z 308[M+1] + .

[0156] Step 9: Synthesize compound 2J

[0157] DMF (5 mL) was added to a flask, followed by compound 2I (0.4 g, 1.31 mmol), intermediate INT-1 (0.31 g, 1.31 mmol), DBU (0.4 g, 2.62 mmol), and BOP (0.87 g, 1.97 mmol). The mixture was stirred at 60 °C for 18 h. After the reaction was complete as shown by TLC, the mixture was diluted with ethyl acetate and added to a saturated NH4Cl aqueous solution (50 mL). The aqueous phase was separated and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified using a preparative silica gel plate (electrolyte: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 2J (158 mg, pale yellow solid, 23% yield).

[0158] MS(ESI): m / z 524[M+1] + .

[0159] Step 10: Synthesize Compound 2

[0160] Compound 2J (100 mg, 0.2 mmol) was added to methanol (5 mL) at room temperature, followed by 10% palladium on carbon (20 mg). After the addition was complete, a hydrogen balloon was attached to the bottle mouth, and the gas was purged three times. The reaction was then carried out at room temperature under a hydrogen atmosphere for 3 h. After the reaction was completed by TLC, the mixture was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was removed by vacuum distillation, and the residue was purified by preparative silica gel plate (electrolyte: petroleum ether / ethyl acetate = 1:1 (v / v)) to give compound 2 (66 mg, gray solid, yield 67%).

[0161] MS(ESI): m / z 494[M+1] + .

[0162] 1H-NMR (400MHz, CDCl3): δ7.27-7.20(m,2H),7.18-7.10(m,3H),6.96-6.85(m,2H),6.76(s,1H),6.71-6.66(m,2H),6.18-5. 70(m,2H),5.48-5.37(m,1H),4.50-4.37(m,2H),4.29-4.18(m,2H),3.36-3.29(m,2H),2.40(s,3H),1.47(d,J=4.0Hz,3H).

[0163] Example 3: Preparation of Compound 3

[0164]

[0165] Synthesis route:

[0166]

[0167] Preparation method:

[0168] Step 1: Synthesize compound 3A

[0169] Compound 2E (2 g, 7.35 mmol) was dissolved in DCM (20 mL), acetic anhydride (1.78 g, 11.03 mmol) was added, followed by DIEA (3.8 g, 29.5 mmol), and the mixture was stirred and refluxed overnight at 45 °C. After the reaction was completed by TLC monitoring, the mixture was added to a saturated NH4Cl aqueous solution (100 mL), and the aqueous phase was separated and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, dissolved under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1 (v / v)) to give compound 3A (2.4 g, yield 82.3%).

[0170] MS(ESI): m / z 314,316[M+1] + .

[0171] Step 2: Synthesize compound 3B

[0172] Compound 3A (1.9 g, 6.04 mmol) was dissolved in dioxane (20 mL), followed by diphenyl ketone imine (2 g, 12.08 mmol), Pd₂(dba)₃ (0.24 g, 604 μmol), Xantphos (0.35 g, 1.2 mmol), and Cs₂CO₃ (3.9 g, 12.08 mmol). The mixture was stirred and refluxed overnight at 95 °C under nitrogen protection. After the reaction was complete as shown by TLC, the mixture was added to a saturated NH₄Cl aqueous solution (100 mL), and the aqueous phase was separated and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and dissolved under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1 (v / v)) to give compound 3B (2.3 g, 93% yield).

[0173] MS(ESI): m / z 415[M+1] + .

[0174] Step 3: Synthesizing compound 3C

[0175] Compound 3B (2.1 g, 5.0 mmol) was dissolved in methanol (20 mL), and ethyl hydrochloride solution (5 mL, 8% by mass) was added dropwise at low temperature. The mixture was stirred overnight at room temperature. After the reaction was complete as shown by TLC, the solution was evaporated to dryness, slurried with petroleum ether / ethyl acetate (1:1 v / v), and dried to give compound 3C (1.1 g, 90% yield).

[0176] MS(ESI): m / z 251[M+1] + .

[0177] Step 4: Synthesize compound 3D

[0178] Compound 3C (1.0 g, 4.0 mmol) was added to a sealed container containing acetonitrile (20 mL), followed by the addition of ethyl hydrochloride solution (5 mL, 8% by mass). The mixture was stirred overnight at 100 °C. After the reaction was complete as shown by TLC, the mixture was cooled to room temperature, and a solid precipitated. The solid was filtered, washed with cold ethyl acetate, and then dried to give compound 3D (0.7 g, 60% yield).

[0179] MS(ESI): m / z 260[M+1] + .

[0180] Step 5: Synthesize compound 3E

[0181] DMF (5 mL) was added to a flask, followed by compound 3D (0.34 g, 1.31 mmol), intermediate INT-1 (0.31 g, 1.31 mmol), DBU (0.4 g, 2.62 mmol), and BOP (0.87 g, 1.97 mmol). The mixture was stirred at 60 °C for 18 h. After the reaction was complete as shown by TLC, the mixture was diluted with ethyl acetate and added to a saturated NH4Cl aqueous solution (50 mL). The aqueous phase was separated and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified using a preparative silica gel plate (electrolyte: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 3E (205 mg, pale yellow solid, 33% yield).

[0182] MS(ESI): m / z 476[M+1] + .

[0183] Step 6: Synthesize compound 3

[0184] Compound 3E (150 mg, 0.32 mmol) was added to methanol (5 mL) at room temperature, followed by 10% palladium on carbon (20 mg). After the addition was complete, a hydrogen balloon was attached to the bottle mouth, and the gas was purged three times. The reaction was then carried out at room temperature under a hydrogen atmosphere for 3 h. After the reaction was completed by TLC, the mixture was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was removed by vacuum distillation, and the residue was purified using a preparative silica gel plate (electrolyte: petroleum ether / ethyl acetate = 1:1 (v / v)) to give compound 3 (92 mg, gray solid, 65% yield).

[0185] MS(ESI): m / z 446[M+1] + .

[0186] 1 H-NMR (400MHz, DMSO-d6): δ8.45(s,1H),8.17(d,J=7.6Hz,1H),6.97(s,1H),6.88(s,1H),6.84(s,1H),6.69(s,1H),5 .61-5.46(m,3H),4.45-4.28(m,2H),4.11-3.95(m,1H),3.93-3.77(m,1H),2.41-2.29(m,6H),1.53(d,J=7.1Hz,3H).

[0187] Example 4: Preparation of Compound 4

[0188]

[0189] Synthesis route:

[0190]

[0191] Preparation method:

[0192] Step 1: Synthesize compound 4A

[0193] Compound 2E (2 g, 7.35 mmol) was dissolved in DCM (20 mL), and N-methylpiperidin-4-formyl chloride (1.79 g, 11.03 mmol) was added, followed by DIEA (3.8 g, 29.5 mmol). The mixture was stirred and refluxed overnight at 45 °C. After the reaction was completed by TLC monitoring, the solution was added to a saturated NH4Cl aqueous solution (50 mL), and the aqueous phase was separated and extracted with DCM (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, dissolved under reduced pressure, and the residue was purified using a preparative silica gel plate (developing solvent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4A (2.34 g, yield 80.1%).

[0194] MS(ESI): m / z 397,399[M+1] + .

[0195] Step 2: Synthesize compound 4B

[0196] Compound 4A (2.4 g, 6.04 mmol) was dissolved in dioxane (20 mL), followed by diphenyl ketone imine (2 g, 12.08 mmol), Pd₂(dba)₃ (0.24 g, 604 μmol), Xantphos (0.35 g, 1.2 mmol), and Cs₂CO₃ (3.9 g, 12.08 mmol). The mixture was stirred and refluxed overnight at 95 °C under nitrogen protection. After the reaction was complete as shown by TLC, the mixture was added to a saturated NH₄Cl aqueous solution (50 mL), and the aqueous phase was separated and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified using preparative silica gel agar plates (developing solvent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4B (2.74 g, 91% yield).

[0197] MS(ESI): m / z 498[M+1] + .

[0198] Step 3: Synthesize compound 4C

[0199] Compound 4B (2.5 g, 5.0 mmol) was dissolved in methanol (50 mL), and 8% ethyl acetate-HCl was added dropwise at low temperature. The mixture was left to stand overnight at room temperature. After the reaction was complete as shown by TLC, the solution was evaporated to dryness, slurried with petroleum ether / ethyl acetate (1:1), and dried to give compound 4C (1.43 g, 86% yield).

[0200] MS(ESI): m / z 334[M+1] + .

[0201] Step 4: Synthesize compound 4D

[0202] Compound 4C (1.0 g, 3.0 mmol) was added to a sealed container containing acetonitrile (10 mL), followed by the addition of ethyl hydrochloride solution (5 mL, 8% by mass). The mixture was stirred overnight at 100 °C. After the reaction was complete as shown by TLC, the mixture was cooled to room temperature, and a solid precipitated. The solid was filtered, washed with cold ethyl acetate, and then dried to give compound 4D (0.67 g, 65% yield).

[0203] MS(ESI): m / z 343[M+1] + .

[0204] Step 5: Synthesize compound 4F

[0205] DMF (5 mL) was added to a flask, followed by compound 4D (0.45 g, 1.31 mmol), compound 4E (0.31 g, 1.31 mmol), DBU (0.4 g, 2.62 mmol), and BOP (0.87 g, 1.97 mmol). The mixture was stirred at 60 °C for 18 h. After the reaction was complete as shown by TLC, the mixture was diluted with ethyl acetate and added to a saturated NH4Cl aqueous solution (50 mL). The aqueous phase was separated and extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified using a preparative silica gel plate (developing solvent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4F (263 mg, pale yellow solid, yield 36%).

[0206] MS(ESI): m / z 559[M+1] + .

[0207] Step 6: Synthesize compound 4

[0208] Compound 4F (178 mg, 0.32 mmol) was added to methanol (5 mL) at room temperature, followed by 10% palladium on carbon (20 mg). After the addition was complete, a hydrogen balloon was attached to the bottle neck, and the gas was purged three times. The reaction was then carried out at room temperature under a hydrogen atmosphere for 3 h. After the reaction was complete as shown by TLC, the mixture was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was removed by vacuum distillation, and the residue was purified using a preparative silica gel plate (electrolyte: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 4 (108 mg, gray solid, yield 64%).

[0209] MS(ESI): m / z 529[M+1] + .

[0210] 1 H-NMR (400MHz, DMSO): δ8.45(s,1H),8.17(d,J=7.6Hz,1H),6.97(s,1H),6.88(s,1H),6.84(s,1H),6.69(s,1H),5.90-5.78(m,3H),4.45- 4.25(m,2H),4.12-3.80(m,2H),3.05-2.70(m,4H),2.49-2.41(m,1H),2.30(s,3H),2.13(s,3H),1.84-1.64(m,4H),1.59(d,J=7.1Hz,3H).

[0211] Example 5: Preparation of Compound 5

[0212]

[0213] Synthesis route:

[0214]

[0215] Preparation method:

[0216] Step 1: Synthesize compound 5

[0217] DMF (5 mL) was added to a flask, followed by compound 4D (0.45 g, 1.31 mmol), compound 5A (0.25 g, 1.31 mmol), DBU (0.4 g, 2.62 mmol), and BOP (0.87 g, 1.97 mmol). The mixture was stirred at 60 °C for 18 h. After the reaction was complete as shown by TLC, the mixture was added to a saturated NH4Cl aqueous solution (50 mL), and the aqueous phase was separated. The aqueous phase was extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and dissolved under reduced pressure. The residue was purified using a preparative silica gel plate (developing solvent: petroleum ether / ethyl acetate = 1:1 (v / v)) to give compound 5 (263 mg, pale yellow solid, yield 36%).

[0218] MS(ESI): m / z 514[M+1] + .

[0219] 1 H-NMR (400MHz, DMSO): δ8.52(s,1H),8.32(d,J=4.0Hz,1H),7.68(t,J=8Hz,1H) ,7.49(t,J=8Hz,1H),7.27(t,J=8Hz,1H),7.23(t,J=52Hz,1H),6.98(s,1H),5.9 0-5.78(m,1H),4.45-4.25(m,2H),4.12-3.80(m,2H),3.05-2.70(m,4H),2.49-2 .41(m,1H),2.30(s,3H),2.13(s,3H),1.84-1.64(m,4H),1.59(d,J=7.1Hz,3H).

[0220] Example 6: Preparation of compounds 6-1 and 6-2

[0221]

[0222] Synthesis route:

[0223]

[0224] Preparation method:

[0225] Step 1: Synthesize a mixture of compounds 6C-1 and 6C-2

[0226] Compound 6A (46 g, 274 mmol) was dissolved in DMF (500 mL), and potassium carbonate (46 g, 333 mmol) was added at room temperature. Then, compound 6B (86 g, 333 mmol) was added dropwise at room temperature. After the addition was complete, the temperature was raised to 90 °C and the reaction was carried out for 3 h. After the reaction was completed by TLC, the reaction solution was cooled to room temperature and then slowly added to water (2500 mL). The precipitated solid was filtered, washed several times with water, and dried to obtain a mixture of compounds 6C-1 and 6C-2 (52 g, white solid, yield 71%).

[0227] MS(ESI): m / z 267[M+1] + .

[0228] Step 2: Synthesize a mixture of compounds 6D-1 and 6D-2

[0229] A mixture of compounds 6C-1 and 6C-2 (25 g, 93.6 mmol) was added in portions to concentrated sulfuric acid (250 mL) under ice bath conditions. While maintaining the ice bath, potassium nitrate (9.4 g, 93.6 mmol) was slowly added in portions, keeping the internal temperature below 5 °C. After the addition was complete, the ice bath was removed, and the mixture was heated to room temperature and allowed to react for another 2 h. TLC showed that the reaction was complete. The reaction mixture was then slowly added to stirred ice water, keeping the water temperature below 10 °C. A large amount of solid precipitated after the addition. The solid was filtered, washed several times with water, and dried to obtain a mixture of compounds 6D-1 and 6D-2 (26 g, white solid, 89% yield).

[0230] MS(ESI): m / z 312[M+1] + .

[0231] Step 3: Synthesize a mixture of compounds 6E-1 and 6E-2

[0232] A mixture of compounds 6D-1 and 6D-2 (25 g, 80 mmol) was dissolved in a mixed solvent of ethanol (250 mL) and water (50 mL). Iron powder (22 g, 400 mmol) and ammonium chloride (21 g, 400 mmol) were added at room temperature, and the reaction mixture was then heated under reflux overnight. After the reaction was complete as indicated by TLC, the reaction mixture was filtered, evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1:1 (v / v)) to give a mixture of compounds 6E-1 and 6E-2 (19.8 g, yellow solid, 88% yield).

[0233] MS(ESI): m / z 282[M+1] + .

[0234] Step 4: Synthesize a mixture of compounds 6F-1 and 6F-2

[0235] In a 250 mL sealed container, a mixture of compounds 6E-1 and 6E-2 (14.1 g, 50 mmol) was dissolved in 100 mL of 4N hydrochloric acid acetonitrile solution. After sealing, the mixture was heated to 110 °C and reacted for 8 h. TLC showed that after the reaction was complete, the mixture was cooled to room temperature, and a large amount of solid precipitated. The solid was filtered, washed with diethyl ether, and dried to give a mixture of compounds 6F-1 and 6F-2 (11.9 g, white solid, yield 82%).

[0236] MS(ESI): m / z 291[M+1] + .

[0237] Step 5: Synthesize compounds 6-1 and 6-2

[0238] DMF was added to a flask, followed by a mixture of compounds 6F-1 and 6F-2 (0.38 g, 1.31 mmol). Compounds 5A (0.25 g, 1.31 mmol), DBU (0.4 g, 2.62 mmol), and BOP (0.87 g, 1.97 mmol) were then added. The mixture was stirred at 60 °C for 18 h. After the reaction was complete as shown by TLC, the mixture was diluted with ethyl acetate, extracted with water, and dried to obtain a mixture of compounds 6F-1 and 6F-2. This mixture was then separated by prep-TLC (electrolyte: petroleum ether / ethyl acetate = 3:1 (v / v)) to obtain the highly polar component (25 mg) and the less polar component (22 mg).

[0239] Small polarity components:

[0240] MS(ESI): m / z 462[M+1] + .

[0241] 1 H NMR (400MHz, CDCl3) δ7.55–7.49(m,2H),7.39–7.34(m,1H),7.25(d,J=4.4Hz,1H),7.19(t,J=7.7Hz,1H),6.92(t,J=55.0Hz,1H), 5.77(s,J=7.0Hz,1H),4.92–4.84(m,1H),4.55–4.41(m,2H),4.34–4.21(m,2H),2.57–2.45(m,3H),1.71–1.59(m,3H),1.33–1.26

[0242] Highly polar components:

[0243] MS(ESI): m / z 462[M+1] + .

[0244] 1H-NMR(400MHz, CDCl3):1H NMR (400MHz, CDCl3) δ7.57–7.45(m,2H),7.34(s,1H),7.20(n,2H),6.92(t,J=55.1,1H),5.77(n,1H),5.74–5.67(m,1H ),4.94(t,J=3.3Hz,1H),4.57–4.39(m,2H),4.33–4.21(m,2H),2.51(s,3H),1.68(d,J=6.7Hz,3H),1.30–1.26(n,3H).

[0245] Verification of activity

[0246] Experiment Example 1: Experiment combining KRAS (G12C) and SOS1

[0247] This assay can be used to examine the efficacy of compounds in inhibiting protein-protein interactions between SOS1 and KRAS G12C. Lower IC50 values ​​indicate a lower efficacy threshold. 50 The value indicates the potency of the compound as an SOS1 inhibitor in the following assay settings.

[0248] 1. Experimental materials:

[0249] The KRAS(G12C) protein was synthesized by Pujian Biotechnology Co., Ltd.

[0250] SOS1 protein exchange human recombinant domain protein (564-1049) was purchased from Cytoskeleton;

[0251] The monoclonal antibody against XL665 tagged with 6-histidine (Mab Anti 6HIS-XL665) and the monoclonal antibody against europium cryptate tagged with glutathione thiol transferase (Mab Anti GST-Eu cryptate) were purchased from Cisbio.

[0252] 2. Experimental Methods:

[0253] 1X buffer preparation (prepare fresh): Hepes: 5mM; NaCl: 150mM; EDTA: 10mM; Igepal: 0.0025%; KF: 100mM; DTT: 1mM; BSA: 0.005%.

[0254] The test compound was diluted 3-fold to the 8th concentration using a multi-channel pipette, i.e., from 100 μM to 45.7 nM.

[0255] Dilute each analyte with 1X buffer to prepare a working solution of 2% DMSO. Add 5 μL / well to the corresponding well and set up a duplicate well experiment. Centrifuge at 1000 rpm for 1 min.

[0256] Prepare a mixed working solution of KRAS(G12C) (200 nM) and Mab Anti GST-Eu cryptate (1 ng / μL) using 1X buffer. Incubate the mixed working solution at 25°C for 5 min and add 2.5 μL / well to the corresponding well.

[0257] A mixed working solution of SOS1 (80 nM) and Mab Anti 6HIS-XL665 (8 g / μL) was prepared using 1X buffer. 2.5 μL / well was added to the corresponding well. 2.5 μL of Mab Anti 6HIS-XL665 (8 g / μL) dilution buffer was added to the Blank well. The final concentration gradient of the compounds was: 1 μM diluted to 0.457 nM, KRAS (G12C) (500 nM), MAb Anti GST-Eu cryptate (0.25 ng / μL), SOS1 (20 nM), and Mab Anti 6HIS-XL665 (2 g / μL). The reaction system was incubated at 25 °C for 60 min. After the reaction, the HTRF was read using a multilabel analyzer.

[0258] 3. Data Analysis:

[0259] The raw data were converted into inhibition rate (IC) using the equation (sample - Min) / (Max - Min) × 100%. 50 The values ​​can be obtained by curve fitting using four parameters (obtained in GraphPad Prism using log(inhibitor) vs. response -- Variable slope mode). Table 1 provides the inhibitory activity of the compounds of this invention on the binding of KRAS(G12C) and SOS1.

[0260] Max well: 1% DMSO, KRAS (G12C) (500nM), MAb Anti GST-Eu cryptate (0.25ng / μL), SOS1 (20nM), Mab Anti 6HIS-XL665 (2g / μL)

[0261] Min well: 1% DMSO, KRAS (G12C) (500nM), MAb Anti GST-Eu cryptate (0.25ng / μL), Mab Anti 6HIS-XL665 (2g / μL)

[0262] Table 1. IC50 values ​​of the compounds of the present invention for inhibiting the binding of KRAS(G12C) and SOS1. 50 data

[0263] Compound numbering <![CDATA[IC 50 (nM)]]> Reference compound BI-3406 19.4 Compound 1 403.4 Compound 2 1233.0 Compound 3 122.4 Compound 4 9.2 Compound 5 20.0 The highly polar component in the mixture of compounds 6-1 and 6-2 >3000 Small polar components in the mixture of compounds 6-1 and 6-2 >3000

[0264] As shown in Table 1, the compounds of the present invention have a good inhibitory effect on SOS1 and a significant inhibitory effect on the binding of KRAS(G12C) and SOS1, which has a good prospect for clinical application.

[0265] Experiment Example 2: Cell Proliferation Inhibition Experiment

[0266] Cell proliferation inhibition assays were used to examine the efficacy of compounds in in vitro in inhibiting SOS1-mediated proliferation, growth, and apoptosis in cancer cell lines. Lower IC50 values ​​were observed. 50 The values ​​represent the high potency of the compounds acting as SOS1 inhibitors in the following assay settings. Specifically, compounds acting as SOS1 inhibitors were observed to effectively inhibit the proliferation of KRAS mutant human cancer cell lines, while KRAS wild-type human cancer cell lines did not show effective inhibition. This confirms the molecular mechanism of action of compounds acting as SOS1 inhibitors, selectively targeting cancer cells dependent on the function of RAS family proteins.

[0267] 1. Experimental materials:

[0268] RPMI 1640 medium, fetal bovine serum, and penicillin / streptomycin antibiotics were purchased from Vicente.

[0269] Low-melting-point agarose was purchased from Sigma;

[0270] Almar blue reagent was purchased from Invitrogen;

[0271] The NCI-H358 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd.

[0272] The Nivo multi-label analyzer was purchased from Perkin Elmer.

[0273] 2. Experimental Methods:

[0274] H358 cells were seeded in 96-well U-shaped plates. A 2% stock solution of low-melting-point agarose was prepared. Before use, the agarose stock solution was heated in a microwave oven until completely melted, then placed in a 42°C water bath to keep the agarose in a liquid state. The gel was added to serum-containing culture medium to prepare a 0.6% gel concentration as the bottom layer, and layered into 96-well U-shaped plates at a density of 50 μL / well. After the bottom layer solidified, the 2% gel was added to cell-containing culture medium to prepare a 0.4% gel concentration as the top layer, with a cell density of 4 x 10⁻⁶ cells / well. 4Cells / mL: Add 75 μL / well to 96-well U-shaped plates coated with the base layer gel, resulting in a cell density of 3000 cells / well. After the top layer gel solidifies, incubate the cell culture plates overnight in a CO2 incubator.

[0275] On the day of compound addition, 85 μL of liquid culture medium was added to a 96-well U-shaped plate containing cells. The test compound was then diluted 3-fold to the 9th concentration using a pipette, from 6 mM to 0.9 μM, for a double-duplicate assay. 97 μL of culture medium was added to the intermediate plate, and then 2.5 μL / well of the serially diluted compound was transferred to the intermediate plate according to the corresponding positions. After mixing, 40 μL / well was transferred to the cell plate. The concentration range of the compound transferred to the cell plate was 30 μM to 4.5 nM. The cell plate was incubated in a CO2 incubator for 7 days. On day 8, the test compound was diluted 3-fold to the 9th concentration using a pipette, from 6 mM to 0.9 μM, for a double-duplicate assay. Add 198 μL of culture medium to the intermediate plate. Then, according to the corresponding positions, transfer 2 μL / well of serially diluted compound to the first intermediate plate. Add 100 μL of culture medium to the second intermediate plate, and add 100 μL of the mixed compound from the first intermediate plate. After mixing, transfer 40 μL / well to the cell plate. The concentration range of the compound transferred to the cell plate is 30 μM to 4.5 nM. Incubate the cell plates in a CO2 incubator for another 7 days. Co-incubate the compound and cells for 14 days. Add 20 μL / well of Almar blue assay reagent to the cell plate, place the dyed plate on a horizontal shaker and shake for 15 min, then incubate the plate at room temperature for 5 h to allow the luminescence signal to stabilize. Read the values ​​using a multi-label analyzer.

[0276] 3. Data Analysis:

[0277] Using the equation (sample - Min) / (Max - Min) * 100%, the raw data are converted into inhibition rate, IC50. 50 The values ​​can be obtained by curve fitting using four parameters (in GraphPad Prism, in the "log(inhibitor) vs. response -- Variable slope" mode). Table 2 provides the inhibitory activity of the compounds of this invention on the proliferation of NCI-H358 cells. ND indicates no test.

[0278] Table 2. IC50 of the compounds of the present invention on the proliferation inhibition of NCI-H358 cells 50 data

[0279] Compound numbering <![CDATA[IC 50 (nM) / NCI-H358]]> Reference compound BI-3406 50.76 Compound 1 585 Compound 2 ND Compound 3 ND Compound 4 ND Compound 5 ND The highly polar component in the mixture of compounds 6-1 and 6-2 ND Small polar components in the mixture of compounds 6-1 and 6-2 ND

[0280] As shown in Table 2, the compounds of the present invention have good anti-proliferative effects against human non-small cell lung cancer NCI-H358, with cell activity less than 1 μM, and have good prospects for clinical application.

[0281] Experiment Example 3: p-ERK Experiment

[0282] 1. Experimental materials:

[0283] DLD-1 cells were purchased from Wuhan Pronosai Life Science Technology Co., Ltd.; 1640 culture medium was purchased from Biological Industries; fetal bovine serum was purchased from Biosera; and Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT was purchased from Cisbio.

[0284] 2. Experimental Methods:

[0285] DLD-1 cells were seeded in clear 96-well cell culture plates with 80 μL of cell suspension per well, containing 8000 DLD-1 cells per well. The cell culture plates were placed in a CO2 incubator and incubated overnight at 37°C.

[0286] The test compound was diluted to 2 mM with 100% DMSO as the first concentration, and then diluted 5-fold to the eighth concentration, i.e., from 2 mM to 0.026 μM, using a pipette. 2 μL of the compound was added to 78 μL of cell starvation medium, mixed well, and then 20 μL of the compound solution was added to the corresponding well of the cell plate. The cell plate was returned to the CO2 incubator for another hour. At this point, the compound concentration was 10 μM to 0.128 nM, and the DMSO concentration was 0.5%.

[0287] After incubation, discard the cell supernatant and add 50 μL of cell lysis buffer to each well, then incubate at room temperature with shaking for 30 minutes.

[0288] Phospho-ERK1 / 2Eu Cryptate antibody and Phospho-ERK1 / 2d2 antibody were diluted 20-fold using detection buffer;

[0289] Take 16 μL of cell lysate supernatant into each well of a new 384 white microplate, then add 2 μL of Phospho-ERK1 / 2Eu Cryptate antibody dilution buffer and 2 μL of Phospho-ERK1 / 2d2 antibody dilution buffer, and incubate at room temperature for 4 hours.

[0290] After incubation, HTRF excitation was read at 320 nm and emission at 615 nm and 665 nm using a multi-label analyzer.

[0291] 3. Data Analysis:

[0292] The original data is converted into inhibition rate using the equation (Sample-Min) / (Max-Min)*100%. The IC50 value can then be obtained by curve fitting using four parameters (obtained in GraphPad Prism using log(inhibitor) vs. response -- Variable slope mode). ND indicates no test was conducted.

[0293] Table 3. IC50 of the compounds of the present invention inhibiting phosphorylation in DLD-1 cells 50 data

[0294] Compound numbering <![CDATA[IC 50 (nM)]]> Reference compound BI-3406 20.0 Compound 1 ND Compound 2 ND Compound 3 402.4 Compound 4 35.4 Compound 5 20.7 The highly polar component in the mixture of compounds 6-1 and 6-2 ND Small polar components in the mixture of compounds 6-1 and 6-2 ND

[0295] As shown in Table 3, the compounds of the present invention, such as compounds 4 and 5, have a good inhibitory effect on phosphorylation of DLD-1 cells and have good prospects for clinical application.

[0296] Although embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Without departing from the principles and spirit of the present invention, those skilled in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention, and all such changes, modifications, substitutions, and variations are covered within the scope of the present invention.

Claims

1. A compound as shown in Formula I or a pharmaceutically acceptable salt thereof, in, A is a phenyl group, wherein the phenyl group is substituted by one R6 molecule; R6 is a C1-C6 alkyl group, wherein the alkyl group is substituted by at least one R8; Each R8 is independently selected from chlorine and fluorine; X and Y are CH; Q1 is selected from -O- and -NR9-; If present, R9 is selected from -C(=O)-R7 and C1-C6 alkyl groups, wherein the alkyl group is separated by one C6-C 10 Aryl substitution; If present, R7 is selected from C1-C6 alkyl groups and 3 to 14-membered heterocyclic alkyl groups, wherein the heterocyclic alkyl group is substituted with one methyl group; Q2 is -O-; L1 and L2 are -(CH2) m -, where m is 1; R1 and R2 are each independently selected from hydrogen and methyl; R3 is a methyl group; R4 is selected from halogens and -NH2; R5 is selected from hydrogen and -C(=O)-OC1-C6 alkyl groups.

2. The compound according to claim 1, characterized in that, It is a compound as shown in Formula I-1. in, q is 1; X, Y, R1, R2, R3, R4, R5 and R6 are as defined in claim 1.

3. The compound according to claim 1 or 2, characterized in that, It is a compound as shown in Formula I-1-2. in, q is 1; R7 is a C1-C6 alkyl group; X, Y, R1, R2, R3, R4 and R6 are as defined in claim 1.

4. The compound according to claim 1, characterized in that, It is a compound as shown in Formula I-3-1. in, q is 1; X, Y, R1, R2, R3, R4, R6 and R9 are as defined in claim 1.

5. The compound according to claim 4, characterized in that, R9 is selected from any one of the following groups: 。 6. The compound according to claim 1, characterized in that, It is selected from the following compounds: 。 7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

8. The use of the compound of any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 7, in the preparation of a medicament for the prevention and / or treatment of diseases or conditions caused by SOS1 overexpression.

9. The application according to claim 8, characterized in that, The disease or condition caused by SOS1 overexpression is cancer.

10. The application according to claim 9, characterized in that, The cancers mentioned are selected from pancreatic cancer, colorectal cancer, and lung cancer.

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