CN115260207A - Tricyclic compound, pharmaceutical composition and application thereof - Google Patents

Abstract

The invention belongs to the field of medicinal chemistry, and relates to a tricyclic compound, a medicinal composition and application thereof. Particularly, the invention relates to a tricyclic compound shown as a formula I, a pharmaceutical composition containing the tricyclic compound and application of the tricyclic compound as an SOS1 inhibitor in the field of medicines. The tricyclic compound shows excellent biological activity and can become a drug property, and has great drug development prospect.

Description

Tricyclic compound and its pharmaceutical composition and application

technical field

The invention belongs to the field of medicinal chemistry, and in particular relates to a class of trihecyclic compounds, a pharmaceutical composition containing the compound and its application in the field of medicine.

Background technique

Since the discovery in late 1982 that RAS family GTPases (KRAS, NRAS, and HRAS) are associated with cancer, the incidence of human cancers is as high as 20% to 30%. RAS proteins act as molecular switches that cycle between an active GTP-bound state and an inactive GDP-bound state. Activated by guanine nucleotide exchange factors (GEFs), the RAS in its GTP-bound state interacts with a number of effectors. Return to the inactive state is driven by GTPase activating proteins (GAPs), which downregulate active RAS by accelerating weak intrinsic GTPase activity by up to 5 orders of magnitude.

Whether mutant RAS proteins require GEF activity for full activation remains to be fully investigated and may vary for specific mutations. The most studied RAS sevenless son (SOS) protein, two known human isoforms, SOS1 and SOS2. Attempts to mimic the RAS-SOS interaction with nanomolar affinity ortho-SOS helix recognition hydrocarbon-stapled peptides by inhibitory peptides have only low cellular activity. Fragment-based screening, rational design, and high-throughput screening methods led to the identification of small molecules addressing the KRAS-SOS1 interaction, resulting in moderate micromolar affinities.

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), pleckstrin ) homology domain (PH), helix linker (HL), RAS exchange motif (REM), CDC25 homology domain and C-terminal proline-rich domain (PR). SOS1 has two binding sites for RAS family proteins; a catalytic site, which binds GDP-bound RAS family proteins to facilitate guanine nucleotide exchange; and an allosteric site, which binds GTP binds RAS family proteins, which leads to a further increase in the catalytic GEF function of SOS1. Published data suggest that SOS1 is critically 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 rate and survival of tumor cells harboring KRAS mutations, whereas no effect was observed in KRAS wild-type cell lines. The effects of SOS1 loss could not be compensated by the introduction of SOS1 with catalytic site mutations, demonstrating the essential role of SOS1GEF activity in KRAS mutant cancer cells.

SOS1 is critically involved in the activation of RAS family protein signaling in cancer through mechanisms other than RAS family protein mutation. SOS1 interacts with the adapter 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 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 RAS family protein activation. SOS1 activation by RAS family proteins may also be mediated through the interaction of SOS1/Grb2 with the BCR-ABL oncoprotein commonly found in chronic myeloid leukemia.

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

In this paper, we describe novel SOS1 inhibitor compounds that bind to the SOS1 catalytic site and simultaneously prevent interaction with RAS family proteins and their activation. This resulted in a marked inhibition of the interaction of SOS1 with RAS family proteins, especially KRAS (with low single digit nanomolar IC50 activity), and thus markedly reduced ERK phosphorylation in KRAS mutant cancer cell lines.

The selective SOS1 inhibitor compounds described herein are expected to provide pharmacological benefit to patients suffering from cancers associated with dependence on RAS family protein signaling. Such cancers that are expected to be targeted by SOS1 inhibitor compounds include those that exhibit alterations (mutation, gene amplification, overexpression) in components (proteins, genes) in the RAS family protein pathway, such as KRAS , NRAS, HRAS, receptor tyrosine kinases (eg, 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 (eg, NF1) and SOS1. Additionally, given the role of SOS1 in RAC1 activation, cancers that exhibit dependence on RAC1 are expected to be targeted by SOS1 inhibitor compounds. In addition, SOS1 inhibitors are expected to be effective in other diseases associated with dysregulation of RAS family protein pathways, such as neurofibromatosis, Noonan syndrome (NS), cardiofaciocutaneous syndrome (CFC) and hereditary gingival fibromatosis type 1. Compounds will also provide pharmacological benefits.

In addition to inhibition and potency, the compounds disclosed herein exhibit good solubility, excellent DMPK profile and good selectivity for kinases of the human kinome.

Contents of the invention

The problem to be solved by the invention

The present invention aims to provide a class of novel structural trihecyclic compounds used as SOS1 inhibitors, which exhibit good inhibitory activity on tumor cells, have good druggability, and have broad prospects for drug development.

solutions to problems

In a first aspect, the present invention provides a compound as shown in formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, wherein

A is selected from C ₆ -C ₁₀ aryl, 5 to 6 membered monocyclic heteroaryl and 9 to 10 membered bicyclic heteroaryl, the aryl, monocyclic heteroaryl and bicyclic heteroaryl are each optionally is substituted by up to 5 R ₆ ;

X and Y are each independently selected from CR ₆ and N;

Q ₁ and Q ₂ are each independently selected from -O-, -C(R ₉ ) ₂ - and -NR ₉ -;

L ₁ and L ₂ are each independently selected from -(CH ₂ ) _m -or -(CH ₂ ) _m -O-(CH ₂ ) _p -O-(CH ₂ ) _n -, wherein each of m, n and p is each independently any integer from 0 to 8;

R ₁ and R ₂ are each independently selected from hydrogen and C ₁ -C ₈ alkyl; or R ₁ and R ₂ form C ₃ -C ₆ cycloalkylene together with the carbon atom to which they are attached;

R ₃ is selected from hydrogen, halogen, cyano, hydroxyl, amino, -NH(R ₆ ), -C(=O)-NH(R ₆ ), C ₁ -C ₆ alkyl, C ₂ -C ₄ alkenyl , C ₂ -C ₄ alkynyl, C ₃ -C ₆ cycloalkyl, 3 to 8 membered heterocycloalkyl, C ₁ -C ₃ alkoxy and C ₁ -C ₆ haloalkyl, the alkyl, alkene Each of radical, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy and haloalkyl is optionally substituted by at least 1 R ₆ ;

R ₄ is selected from hydrogen, halogen, cyano, hydroxyl, amino, -NH(R ₆ ), -C(=O)-NH(R ₆ ), C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkane group, 3 to 8 membered heterocycloalkyl group, C ₁ -C ₃ alkoxy group and C ₁ -C ₆ haloalkyl group, each of the alkyl group, cycloalkyl group, heterocycloalkyl group, alkoxy group and haloalkyl group is any optionally substituted by at least 1 R ₆ ;

R ₅ is selected from hydrogen, halogen, cyano, hydroxyl, amino, -N(R ₆ )(R ₇ ), -C(=O)-N(R ₆ )(R ₇ ), -C(=O)- R ₇ , -C(=O)-OR ₇ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 3 to 8 membered heterocycloalkyl, C ₁ -C ₃ alkoxy and C ₁ -C _Haloalkyl , each of said alkyl, cycloalkyl, heterocycloalkyl, alkoxy and haloalkyl is optionally substituted by at least 1 R _;

If present, each R ₆ and R ₇ is independently selected from hydrogen, halogen, cyano, hydroxyl, amino, carbamoyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ heteroalkyl, C ₃ - C ₈ cycloalkyl, 3 to 14 membered heterocycloalkyl, C ₁ -C ₃ alkoxy, C ₁ -C ₃ haloalkoxy, C ₆ -C ₁₀ aryl, 5 to 6 membered monocyclic heteroaryl and 9 to 10 membered bicyclic heteroaryl, the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, haloalkoxy, aryl, monocyclic heteroaryl and bicyclic heteroaryl Each of the radicals is optionally substituted by at least 1 R ₈ ; or R ₆ and R ₇ together form a 5 to 6 membered heterocycloalkyl group optionally substituted by at least 1 R ₈ replaced;

If present, each R is _{independently} selected from hydrogen, chloro, fluoro, cyano, hydroxy, amino, isopropyl, cyclopropyl, methyl, difluoromethyl, trifluoromethyl, methoxy, Trifluoromethoxy, ethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy and phenyl;

If present, each R ₉ is independently selected from hydrogen, halogen, cyano, hydroxyl, amino, -N(R ₆ )(R ₇ ), -C(=O)-N(R ₆ )(R ₇ ) , -C(=O)-R ₇ , -C(=O)-OR ₇ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 3 to 8 membered heterocycloalkyl, C ₁ - C ₃ alkoxy and C ₁ -C ₆ haloalkyl, said alkyl, cycloalkyl, heterocycloalkyl, alkoxy and haloalkyl are each optionally substituted with at least 1 R ₆ .

Preferably, the compound shown in formula I is the compound shown in formula I-1, wherein

q is any integer from 0 to 4; X, Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are as defined in Formula 1.

More preferably, the compound shown in formula I or formula I-1 is a compound shown in formula I-1-1 or formula I-1-2, wherein

q is any integer from 0 to 4; X, Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R ₇ are as defined in Formula 1.

Further preferably, in the compound shown in formula I-1-1,

基团选自下列基团中的任意一种：

The group is selected from any one of the following groups:

Preferably, the compound shown in formula I is the compound shown in formula I-2, wherein

q is any integer from 0 to 4; X, Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ 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

q is any integer from 0 to 4; X, Y, R ₁ , R ₂ , R ₃ , R ₄ and R ₆ are as defined in formula 1.

Preferably, the compound shown in formula I is the compound shown in formula I-3, wherein

q is any integer from 0 to 4; X, Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₉ 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

q is any integer from 0 to 4; X, Y, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R ₉ are as defined in Formula 1.

Further preferably, in the compound shown in formula I-3-1,

_R9 is selected from any one of the following groups:

In a second aspect, the present invention provides 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 Specific compounds selected from:

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

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

1-(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-ethanone;

(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)methanone;

(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)methanone;

4-(((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-7,8-dihydro-[1,4]di Ethyl oxino[2,3-g]quinazoline-7-carboxylate; and

4-(((R)-1-(3-(Difluoromethyl)-2-fluorophenyl)ethyl)amino)-2-methyl-7,8-dihydro-[1,4]di Ethyl oxino[2,3-g]quinazoline-8-carboxylate.

In a third aspect, the present invention provides a pharmaceutical composition comprising formula I, formula I-1, formula I-1-1, formula I-1-2, formula I-2, formula I-2-1 , the compound shown in formula I-3 or formula I-3-1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug, and at least A pharmaceutically acceptable excipient.

In a fifth aspect, the present invention provides 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 - The compound shown in 3-1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug or a pharmaceutical composition containing it, which is used as SOS1 inhibitors are alternatively useful in the prevention and/or treatment of diseases or conditions caused by overexpression of SOS1.

In a sixth aspect, the present invention provides 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 -The compound shown in 3-1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug or the pharmaceutical composition containing it is used in the preparation of Use in medicines for preventing and/or treating diseases or conditions caused by overexpression of SOS1.

In the seventh aspect, the present invention provides a method for preventing and/or treating diseases or disorders caused by overexpression of SOS1, which includes preventing and/or treating effective doses such as formula I, formula I-1, formula The compound shown in I-1-1, formula I-1-2, formula I-2, formula I-2-1, formula I-3 or formula I-3-1 or its pharmaceutically acceptable salt, hydrated The compound, solvate, stereoisomer, tautomer, metabolite or prodrug, or a pharmaceutical composition comprising the same, is administered to an individual in need thereof.

Preferably, in the above medical use, the disease or condition caused by the overexpression of SOS1 is cancer, preferably pancreatic cancer, colorectal cancer and lung cancer.

The effect of the invention

The present invention provides a series of novel three-apex ring compounds. It is proved by related enzyme and cell activity tests that the compounds of the present invention have excellent cell proliferation inhibitory activity. In vitro experiments, the IC ₅₀ value for cell proliferation reaches nM level, and can be well applied in a variety of tumors. At the same time, the compound of the present invention has a very good inhibitory effect on KRAS:SOS1 activation, which can reach the nM level, and is suitable for being prepared as a SOS1 inhibitor for preventing and/or treating diseases or diseases related to SOS1 activation, such as cancer ( including but not limited to pancreatic cancer, colorectal cancer and lung cancer).

Detailed ways

General Terms and Definitions

Unless stated to the contrary, the terms used in the present invention have the following meanings.

"Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight chain and branched chain groups of 1 to 20 carbon atoms, such as 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 Carbon atoms, straight chain and branched chain groups of 1 to 6 carbon atoms or 1 to 4 carbon atoms. In the present invention, "alkyl" may 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-dimethylpropane base, 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 its various branched isomers, etc. Non-limiting examples also include, but are not limited to, methylene, methine, ethylene, ethylene, propylene, propylene, butylene, butylene, and various branched isomers thereof. In addition, in the present invention, "alkyl" may be optionally substituted or unsubstituted.

"Alkoxy" means an "-O-alkyl" group in which "alkyl" is as defined above.

"Alkenyl" refers to unsaturated aliphatic hydrocarbon groups, including 1 to 20 carbon atoms and at least 1 carbon-carbon double bond straight chain and branched groups, such as 1 to 18 carbon atoms, 1 to Straight chain and branched chain groups of 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms or 1 to 4 carbon atoms. In the present invention, "alkenyl" may be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, vinyl (-CH=CH ₂ ), propen-1-yl (-CH=CH-CH ₃ ), propen-2-yl (-C(CH ₃ )=CH ₂ ), Buten-1-yl (-CH=CH-CH ₂ -CH ₃ ), buten-2-yl (-C(C ₂ H ₅ )=CH ₂ ), 1-methylpropen-1-yl (- C(CH ₃ )=CH-CH ₃ ) and its various branched isomers, etc. Non-limiting examples also include, but are not limited to, 1,1-ethenylene (=C=CH ₂ ), 1,2-ethenylene (-CH=CH-), 1,1-propenylene (=C=CH- CH-CH ₃ ), 1,2-propenylene (-CH=C(CH ₃ )-), 1,3-propenylene (-CH=CH-CH ₂ -) and various branched chain isomers body. In addition, in the present invention, "alkenyl" may be optionally substituted or unsubstituted.

戊炔基

及其各种支链异构体等。非限制性实例还包括但不限于亚乙炔基(-C≡C-)、亚丙炔基

亚丁炔基

及其各种支链异构体。另外，在本发明中，“炔基”可以是任选取代的或未取代的。"Alkynyl" refers to unsaturated aliphatic hydrocarbon groups, including 1 to 20 carbon atoms and at least 1 carbon-carbon triple bond straight chain and branched groups, such as 1 to 18 carbon atoms, 1 to Straight chain and branched chain groups of 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms or 1 to 4 carbon atoms. In the present invention, "alkynyl" may be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, ethynyl (-C≡CH), propynyl (-C≡C- _CH3 ), butynyl

pentynyl

And its various branched chain isomers. Non-limiting examples also include, but are not limited to, ethynylene (-C≡C-), propynylene

Butynylene

and its various branched isomers. In addition, in the present invention, "alkynyl" may be optionally substituted or unsubstituted.

"Heteroalkyl" refers to a saturated aliphatic hydrocarbon group, including straight chain and branched chain groups of 2 to 20 atoms, such as 2 to 18 atoms, 2 to 12 atoms, 2 to 8 atoms, Straight-chain and branched chain groups of 2 to 6 atoms or 2 to 4 atoms, wherein one or more atoms are _hetero atoms, the rest is carbon. In the present invention, "heteroalkyl" may be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, methoxymethyl (2-oxapropyl), methylthiomethyl (2-thiapropyl), methylaminomethyl (2-azapropyl), and various Branched chain isomers, etc. In addition, in the present invention, "heteroalkyl" may be optionally substituted or unsubstituted.

"Cycloalkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group, including 3 to 12 ring atoms, such as 3 to 12, 3 to 10 or 3 to 6 Ring atoms (ie 3 to 6 membered rings). Non-limiting examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclo Heptatrienyl, cyclooctyl, etc. In the present invention, "cycloalkyl" may be optionally substituted or unsubstituted.

"Heterocycloalkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group, including 3 to 20 ring atoms, such as 3 to 16, 3 to 12, 3 to 10 or 3 to 6 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is 0, 1 or 2), and the remaining ring atoms are carbon. Preferably heterocycloalkyl comprises 3 to 12 ring atoms, of which 1 to 4 are heteroatoms, more preferably comprises 3 to 10 ring atoms, most preferably comprises 5 or 6 ring atoms, of which 1 to 4, Preferably 1 to 3, more preferably 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocycloalkyl include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Non-limiting examples of polycyclic heterocycloalkyls include, but are not limited to, spirocyclic or bridged heterocycloalkyls.

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

"Haloalkyl" or "haloalkoxy" refers to an alkyl or alkoxy group substituted by one or more of the same or different halogen atoms, examples of preferred alkyl or alkoxy include but are not limited to: tri Fluoromethyl, trifluoroethyl, trifluoromethoxy.

"Cyano" means a "-CN" group.

"Hydroxy" means a "-OH" group.

"Amino" refers to a " _-NH2 " group.

"Carbamoyl" refers to a "-(C=O) _-NH2 " group.

"Aryl" means monocyclic, bicyclic and tricyclic carbocyclic ring systems containing 6-14 ring atoms, wherein at least one ring system is aromatic, and wherein each ring system contains 3-7 atoms ring with one or more points of attachment to the rest of the molecule. Examples include, but are not limited to: phenyl, naphthyl, anthracene, and the like. Preferably, the aryl group is a carbocyclic ring system of 6-10 or 6-7 ring atoms.

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

"Optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, a "heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes cases where the heterocycle group is substituted with an alkyl group and cases where the heterocycle group is not substituted with an alkyl group .

"Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in a group are independently substituted by a corresponding number of substituents.

"Pharmaceutically acceptable salt" refers to a salt prepared from a compound of the present invention with a relatively non-toxic acid or base. When the compounds of the present invention contain relatively acidic functional groups (such as carboxyl or sulfonic acid groups), base addition salts can be obtained by contacting their free forms with a sufficient amount of base in pure solution or in a suitable inert solvent . 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 compound of the present invention contains a relatively basic functional group (such as amino or guanidino), acid addition salts can be obtained by contacting its free form with a sufficient amount of acid in a pure solution or in a suitable inert solvent . Non-limiting examples of pharmaceutically acceptable acid addition salts include, but are not limited to, inorganic acid salts (e.g., hydrochloride, hydrobromide, hydroiodide, nitrate, carbonate, bicarbonate, phosphate , monohydrogen phosphate, dihydrogen phosphate, phosphite, sulfate, bisulfate, etc.), organic acid salts (such as acetate, propionate, isobutyrate, malonate, succinate , suberate, maleate, fumarate, citrate, tartrate, lactate, mandelate, benzoate, phthalate, methanesulfonate, benzenesulfonate salt, p-toluenesulfonate, glucuronic acid, etc.) and amino acid salts (such as arginine salt, etc.). 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 specific compounds of the present invention contain both basic and acidic functionalities and thus can be converted into either base or acid addition salts. Preferably, the neutral form of the compound is regenerated by contacting the salt with a base or acid in the conventional manner followed by isolation of the parent compound. The parent form of a compound differs from its various salt forms by certain physical properties, such as solubility in polar solvents. According to an embodiment of the present invention, preferably the pharmaceutically acceptable salt of the compound represented by formula I is an acid addition salt, preferably hydrochloride, hydrobromide, phosphate or sulfate, more preferably hydrochloride.

"Pharmaceutical composition" refers to a pharmaceutically acceptable composition comprising one or more compounds of formula I or a pharmaceutically acceptable form thereof (e.g. salt, hydrate, solvate, stereoisomer isomers, tautomers, metabolites, prodrugs, etc.), and other components (such as pharmaceutically acceptable excipients).

In the present invention, "pharmaceutically acceptable excipients" refer to auxiliary materials widely used in the field of pharmaceutical production. The main purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in nature and/or has specific functionality, and also to provide a method so that after the drug is administered to the subject, the active ingredient can be used in the desired manner. The rate of dissolution, or the promotion of effective absorption of the active ingredient in the subject to which it is administered. Pharmaceutically acceptable excipients can be inert fillers, or functional ingredients that provide certain functions for the pharmaceutical composition (such as stabilizing the overall pH value of the composition or preventing the degradation of active ingredients in the composition). Non-limiting examples of pharmaceutically acceptable excipients include, but are not limited to, binders, suspending agents, emulsifying agents, diluents (or fillers), granulating agents, adhesives, disintegrants, lubricants, anti-adherents , glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, sweeteners, etc.

The pharmaceutical compositions of the present invention can be prepared by any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, milling, encapsulating, entrapping and/or lyophilizing processes.

In the present invention, the purpose of using the pharmaceutical composition is to promote the administration to organisms, facilitate the absorption of active ingredients, and then exert biological activity. The pharmaceutical composition of the present invention can be administered by any form, including injection (intra-arterial, intravenous, intramuscular, intraperitoneal, subcutaneous), mucosal, oral (oral solid formulation, oral liquid formulation), rectal, inhalation, implantation , topical (eg ocular) administration, and the like. Non-limiting examples of oral solid formulations include, but are not limited to, powders, capsules, lozenges, granules, tablets, and the like. Non-limiting examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, tinctures, elixirs, solutions, and the like. Non-limiting examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum formulations. Non-limiting examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry powder for injection, suspension for injection, emulsion for injection, and the like. The pharmaceutical composition of the present invention can also be made into controlled-release or delayed-release dosage forms (such as liposomes or microspheres).

Preferably, the compounds of the present invention or pharmaceutical compositions comprising them are administered orally or intravenously to individuals in need thereof. Depending on the specific circumstances of the subject, other routes of administration may also be applicable or even preferred. For example, transdermal administration would be a very important mode of administration for patients who are forgetful or irritable to oral medications. In the present invention, the route of administration can be varied or adjusted in any suitable manner to meet the needs of the nature of the drug, the convenience of the patient and medical staff, and other related factors.

The compounds of the present invention or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, metabolites or prodrugs or pharmaceutical compositions containing them have excellent SOS1 enzyme activity and cell The growth inhibitory activity can be used as an SOS1 inhibitor to prevent and/or treat diseases or diseases caused by the overexpression of SOS1, and has good clinical and medical applications. Preferably, non-limiting examples of diseases or conditions caused by overexpression of SOS1 are cancers, including but not limited to pancreatic cancer, colorectal cancer and lung cancer.

The technical solutions of the present invention will be described below in conjunction with specific examples. The following examples are provided to further illustrate the present invention, but not to limit the scope of the present invention. It will be apparent to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

The preparation of the compounds of the present invention can be achieved by synthetic methods well known to those skilled in the art, including but not limited to the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and the methods described by those skilled in the art. Known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention. The known starting materials used in the present invention can be synthesized by methods known in the art, or purchased through conventional commercial means (for example, purchased from Shaoyuan Chemical Technology, Beijing Coupling Technology, etc.). Unless otherwise specified, the reactions were carried out under argon atmosphere or nitrogen atmosphere. The hydrogenation reaction is usually vacuumized and filled with hydrogen, and the operation is repeated 3 times. The reaction temperature is room temperature, and the temperature range is 20°C-30°C. Monitoring of 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). The thin-layer chromatography silica gel plate uses Qingdao Haiyang GF254 silica gel plate. The developer system includes but 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. adjust.

The separation and purification of the compounds of the present invention can be achieved by synthetic methods well known to those skilled in the art, including but not limited to column chromatography (CC), high performance liquid chromatography (HPLC), ultra high performance liquid chromatography (UPLC) Wait. Column chromatography generally uses Qingdao Haiyang 200-300 mesh silica gel as a carrier. The eluent 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 determined according to the compound. The polarity can be adjusted, and it can also be adjusted by adding a small amount of acidic or alkaline anti-smearing reagent. The HPLC spectrum was determined by Agilent1200DAD HPLC chromatograph (column: Sunfire C18, 150×4.6mm, 5μm) or Waters 2695-2996HPLC chromatograph (column: GiminiC18, 150×4.6mm, 5μm).

The structural identification of the compounds of the present invention can be achieved by methods well known to those skilled in the art, including but not limited to nuclear magnetic resonance (NMR), mass spectrometry (MS) and the like. The NMR spectrum is measured by Bruker AVANCE-400 or Varian Oxford-300 nuclear magnetic apparatus, and the measuring solvent is deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDC1 ₃ ) or deuterated methanol (CD ₃ OD), The internal standard is tetramethylsilane (TMS), and the chemical shift is in 10 ^-6 (ppm). The MS spectrum was determined by Agilent SQD (ESI) mass spectrometer (model: 6110) or Shimadzu SQD (ESI) mass spectrometer (model: 2020).

Preparation of intermediates

Preparation of intermediate INT-1

Step 1: Synthesis of Compound INT-1B

Compound INT-1A (25g, 107mmol) was dissolved in THF (250mL), and tert-butylsulfinamide (19.5g, 161mmol) was added at room temperature, followed by tetraethyl titanate (61g, 267.5mmol). After the addition, the temperature of the reaction solution was raised to 70° C., and the reaction was carried out for 4 hours. After TLC showed that the reaction was over, the reaction solution was cooled to room temperature, and the reaction solution was slowly added to ice water, and the aqueous phase was extracted with ethyl acetate (3×150 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=5/1 (volume ratio)) , to obtain compound INT-1B (27.7 g, pale yellow solid, yield 77%).

MS (ESI): m/z 337 [M+1] ⁺ .

Step 2: Synthesis of Compound INT-1C

Compound INT-1B (27g, 80mmol) was dissolved in a mixed solvent of methanol (200mL) and water (100mL), cooled to -20°C, then sodium borohydride (6g, 160mmol) was added in portions, and the reaction was maintained at -20°C After 1 h, the temperature was naturally raised to room temperature, and the reaction was continued for 3 h at room temperature. TLC showed that after the reaction was completed, the methanol was distilled off under reduced pressure, and the aqueous phase was extracted with ethyl acetate (3×150 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/1 (volume ratio)) , to obtain compound INT-1C (22 g, pale yellow solid, yield 82%).

MS (ESI): m/z 339 [M+1] ⁺ .

Step 3: Synthesis of Compound INT-1

Compound INT-1C (20 g, 59 mmol) was added into 4N HCl-ethyl acetate solution (200 mL), and reacted at room temperature for 2 h. TLC showed that after the reaction was completed, a solid precipitated out, was filtered, and the solid was washed with ethyl acetate and dried to obtain the hydrochloride of compound INT-1 (12.3 g, pale yellow solid, yield 89%). The product was directly used in subsequent reactions without purification.

MS (ESI): m/z 235 [M+1] ⁺ .

Preparation of target compound

Embodiment 1: the preparation of compound 1

synthetic route:

Preparation:

Step 1: Synthesis of Compound 1B

Compound 1A (25g, 104mmol) was dissolved in a mixed solvent of ethanol (250mL) and water (50mL), iron powder (23g, 416mmol) and ammonium chloride (22g, 416mmol) were added at room temperature, and the reaction solution was heated to reflux overnight. After TLC showed that the reaction was over, the reaction solution was filtered, and after the filtrate was spin-dried, the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1:1 (volume ratio)) to obtain compound 1B (16.6 g, yellow solid, yield 76%).

MS (ESI): m/z 210 [M+1] ⁺ .

Step 2: Synthesis of Compound 1C

In a 250 mL stuffy jar, compound 1B (16 g, 76 mmol) was dissolved in 4N hydrochloric acid acetonitrile solution (100 mL), sealed, heated to 110° C., and reacted for 8 h. TLC showed that after the reaction was completed, a large amount of solids were precipitated after cooling to room temperature. The solids were filtered, washed with ether, and dried to obtain compound 1C (13.8 g, white solid, yield 83%).

MS (ESI): m/z 219 [M+1] ⁺ .

Step 3: Synthesis of compound 1D

Compound 1C (10g, 45.6mmol) was added into phosphorus oxychloride (100mL), heated to 120°C, and reacted for 6h. TLC showed that after the reaction was completed, phosphorus oxychloride was distilled off 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). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/1 (volume ratio)) , to obtain compound 1D (8.7 g, pale yellow solid, yield 81%).

MS (ESI): m/z 237 [M+1] ⁺ .

Step 4: Synthesis of compound 1E

Compound 1D (500mg, 2.1mmol) was added to acetonitrile (5mL), then intermediate INT-1 (580mg, 2.5mmol) and DIEPA (540mg, 4.2mmol) were added, after the addition, the reaction solution was heated to 80°C , Reaction 2h. TLC showed that after the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=5:1 (volume ratio)) to obtain compound 1E (590 mg, pale yellow Solid, yield 65.3%).

MS (ESI): m/z 435 [M+1] ⁺ .

Step 5: Synthesis of compound 1

At room temperature, compound 1E (500 mg, 1.1 mmol) was added into methanol (5 mL), and then 10% palladium on carbon (50 mg) was added. After the addition, put a hydrogen balloon on the mouth of the bottle, and after replacing the gas three times, keep the hydrogen atmosphere at room temperature for 3 hours. After TLC showed that the reaction was over, it was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was evaporated under reduced pressure, and the residue was purified on a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=5:1 (volume ratio)) to obtain Compound 1 (300 mg, gray solid, 67% yield).

MS (ESI): m/z 405 [M+1] ⁺ .

¹ H-NMR (400MHz, CDCl ₃ ): 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).

Embodiment 2: the preparation of compound 2

synthetic route:

Preparation:

Step 1: Synthesis of Compound 2B

Compound 2A (50g, 228mmol) was added to a flask containing concentrated sulfuric acid (425mL) under ice-cooling, and KNO ₃ (23.1g, 228mmol) was added in batches under ice-cooling. After stirring for 3 hours, TLC showed that after the reaction was completed, ice water was added to precipitate a solid, which was filtered and dried to obtain compound 2B (49 g, yield 81.6%).

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

Step 2: Synthesis of Compound 2C

Methanol (480mL) was added to a three-necked flask, and compound 2B (49g, 185.6mmol) was added to the flask, and under nitrogen protection, SOCl ₂ (29g, 24.38mmol) was added dropwise under ice bath with stirring. After the addition, room temperature Stir for 10 min, overnight at 70°C. TLC showed that after the reaction was completed, the mixture was spin-dried, slurried with petroleum ether/ethyl acetate (volume ratio 5:1), and dried to obtain compound 2C (43 g, yield 83.5%).

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

Step 3: Synthesis of compound 2D

Dissolve 2-bromoethanol (10.8g, 86.4mmol) in THF (100mL), add lithium triisopropylamide (LDA, 2mol/L) (54mL, 108mmol) dropwise at 0°C, stir at 0°C for 0.5h, Compound 2C (20 g, 72 mmol) was dissolved in THF, and added dropwise to the reaction system, and stirred at room temperature for 3 h. After TLC showed that the reaction was complete, saturated aqueous NH ₄ Cl solution (500 mL) was added, and the aqueous phase was extracted with ethyl acetate (3×150 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/1 (volume ratio)) , to obtain compound 2D (25 g, yield 90.67%).

¹ H-NMR (400MHz, CDCl ₃ ): 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).

Step 4: Synthesis of compound 2E

Compound 2D (25g, 65.2mmol) was dissolved in a mixed solution of ethanol (200mL) and water (50mL), iron powder (11g, 196.5mmol) and ammonium chloride (21.25g, 393mmol) were added, under nitrogen protection, in Stir and reflux overnight at 80°C. TLC showed that after the reaction was completed, it was filtered, and the solid was washed with ethyl acetate several times. The filtrate was added to saturated NH ₄ Cl aqueous solution (500 mL), and the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×150 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/1 (volume ratio)) , to obtain compound 2E (10.8 g, yield 61%).

MS (ESI): m/z 272,274 [M+1] ⁺ .

Step 5: Synthesis of compound 2F

Compound 2E (5g, 18.4mmol) was added to DMF (50mL) at room temperature, then potassium carbonate (5.07g, 26.8mmol) was added, and benzyl bromide (2.82g, 16.56mmol) was added dropwise at room temperature. Reaction 3h. TLC showed that after the reaction was completed, the reaction solution was slowly added to water to precipitate a solid, which was filtered, washed with water, and dried to obtain the gray product 2F (5.2 g, yield 77.6%).

MS (ESI): m/z 362,364 [M+1] ⁺ .

Step 6: Synthesis of Compound 2G

Compound 2F (2.2g, 6.04mmol) was dissolved in dioxane (25mL), diphenylketoneimine (2g, 12.08mmol) was added, Pd ₂ (dba) ₃ (0.24g, 604μmol), xantphos (0.35g, 1.2mmol) and Cs ₂ CO ₃ (3.9g, 12.08mmol) were stirred and refluxed at 95°C overnight under nitrogen protection. TLC showed that after the reaction was completed, the reaction liquid was cooled to room temperature, and added to saturated NH ₄ Cl aqueous solution (100 mL), the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×50 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/1 (volume ratio)) , to obtain compound 2G (3 g, yield 99%).

MS (ESI): m/z 463 [M+1] ⁺ .

Step 7: Synthesis of Compound 2H

Compound 2G (2.8g, 6.03mmol) was dissolved in methanol (25mL) solution, hydrochloric acid ethyl acetate solution (5ml, mass fraction 8%) was added dropwise at low temperature, room temperature overnight. TLC showed that after the reaction was completed, the mixture was spin-dried, slurried with petroleum ether/ethyl acetate (volume ratio 1:1), and dried to obtain compound 2H (1.62 g, yield 90%).

MS (ESI): m/z 299 [M+1] ⁺ .

Step 8: Synthesis of compound 2I

Compound 2H (1.16g, 3.9mmol) was added to a stuffy jar containing acetonitrile (10mL), hydrochloric acid ethyl acetate solution (5ml, mass fraction 8%) was added, and stirred overnight at 100°C. TLC showed that after the reaction was completed, the solid was cooled to room temperature, and the solid was filtered, washed with cold ethyl acetate, and dried to obtain compound 2I (0.7 g, yield 60%).

MS (ESI): m/z 308 [M+1] ⁺ .

Step 9: Synthesis of compound 2J

DMF (5mL) was added to the flask, compound 2I (0.4g, 1.31mmol) was added to the flask, and intermediate INT-1 (0.31g, 1.31mmol), DBU (0.4g, 2.62mmol) and BOP were added (0.87g, 1.97mmol), stirred at 60°C for 18h. TLC showed that after the reaction was completed, it was diluted with ethyl acetate and added to saturated NH ₄ Cl aqueous solution (50 mL), the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×10 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, filtered to remove the desiccant, and precipitated under reduced pressure. The residue was purified with a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=5:1 (volume ratio)) to obtain Compound 2J (158 mg, pale yellow solid, yield 23%).

MS (ESI): m/z 524 [M+1] ⁺ .

Step 10: Synthesis of compound 2

Compound 2J (100 mg, 0.2 mmol) was added into methanol (5 mL) at room temperature, and then 10% palladium on carbon (20 mg) was added. After the addition, put a hydrogen balloon on the mouth of the bottle, and after replacing the gas three times, keep the hydrogen atmosphere at room temperature for 3 hours. After TLC showed that the reaction was over, it was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was evaporated under reduced pressure, and the residue was purified on a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=1:1 (volume ratio)) to obtain Compound 2 (66 mg, gray solid, 67% yield).

MS (ESI): m/z 494 [M+1] ⁺ .

¹ H-NMR (400MHz, CDCl ₃ ): 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).

Embodiment 3: the preparation of compound 3

synthetic route:

Preparation:

Step 1: Synthesis of Compound 3A

Compound 2E (2g, 7.35mmol) was dissolved in DCM (20mL), acetic anhydride (1.78g, 11.03mmol) was added, and then DIEA (3.8g, 29.5mmol) was added, stirred and refluxed at 45°C overnight. After the reaction was monitored by TLC, it was added to saturated NH ₄ Cl aqueous solution (100 mL), and the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×50 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=3/1 (volume ratio)) , to obtain compound 3A (2.4 g, yield 82.3%).

MS (ESI): m/z 314,316 [M+1] ⁺ .

Step 2: Synthesis of compound 3B

Dissolve compound 3A (1.9g, 6.04mmol) in dioxane (20mL), add diphenylketone imine (2g, 12.08mmol), add Pd ₂ (dba) ₃ (0.24g, 604μmol), Xantphos (0.35g, 1.2mmol) and Cs ₂ CO ₃ (3.9g, 12.08mmol) were stirred and refluxed at 95°C overnight under nitrogen protection. TLC showed that after the reaction was completed, it was added to saturated NH ₄ Cl aqueous solution (100 mL), and the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×50 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was precipitated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=5/1 (volume ratio)) , to obtain compound 3B (2.3 g, yield 93%).

MS (ESI): m/z 415 [M+1] ⁺ .

Step 3: Synthesis of Compound 3C

Compound 3B (2.1g, 5.0mmol) was dissolved in methanol (20mL), hydrochloric acid ethyl acetate solution (5ml, mass fraction 8%) was added dropwise at low temperature, and stirred overnight at room temperature. TLC showed that after the reaction was completed, the mixture was spin-dried, slurried with petroleum ether/ethyl acetate (volume ratio 1:1), and dried to obtain compound 3C (1.1 g, yield 90%).

MS (ESI): m/z 251 [M+1] ⁺ .

Step 4: Synthesize Compound 3D

Compound 3C (1.0g, 4.0mmol) was added to a stuffy jar containing acetonitrile (20mL), hydrochloric acid ethyl acetate solution (5ml, mass fraction 8%) was added, and stirred overnight at 100°C. TLC showed that after the reaction was completed, the solid was cooled to room temperature, and the solid was filtered, washed with cold ethyl acetate, and then dried to obtain compound 3D (0.7 g, yield 60%).

MS (ESI): m/z 260 [M+1] ⁺ .

Step 5: Synthesis of compound 3E

DMF (5mL) was added to the flask, compound 3D (0.34g, 1.31mmol) was added to the flask, and intermediate INT-1 (0.31g, 1.31mmol), DBU (0.4g, 2.62mmol) and BOP were added (0.87g, 1.97mmol), stirred at 60°C for 18h. TLC showed that after the reaction was completed, it was diluted with ethyl acetate and added to saturated NH ₄ Cl aqueous solution (50 mL), the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×10 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, filtered to remove the desiccant, and precipitated under reduced pressure. The residue was purified with a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=3:1 (volume ratio)) to obtain Compound 3E (205 mg, pale yellow solid, 33% yield).

MS (ESI): m/z 476 [M+1] ⁺ .

Step 6: Synthesis of compound 3

Compound 3E (150 mg, 0.32 mmol) was added into methanol (5 mL) at room temperature, and then 10% palladium on carbon (20 mg) was added. After the addition, put a hydrogen balloon on the mouth of the bottle, and after replacing the gas three times, keep the hydrogen atmosphere at room temperature for 3 hours. After TLC showed that the reaction was over, it was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was evaporated under reduced pressure, and the residue was purified on a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=1:1 (volume ratio)) to obtain Compound 3 (92 mg, gray solid, 65% yield).

MS (ESI): m/z 446 [M+1] ⁺ .

¹ H-NMR (400MHz, DMSO-d ₆ ): 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).

Embodiment 4: the preparation of compound 4

synthetic route:

Preparation:

Step 1: Synthesis of Compound 4A

Compound 2E (2g, 7.35mmol) was dissolved in DCM (20mL), N-methylpiperidine-4-formyl chloride (1.79g, 11.03mmol) was added, then DIEA (3.8g, 29.5mmol) was added, at 45 °C stirring and reflux overnight. After the reaction was monitored by TLC, it was added to saturated NH ₄ Cl aqueous solution (50 mL), and the aqueous phase was separated, and the aqueous phase was extracted with DCM (3×10 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, filtered to remove the desiccant, and precipitated under reduced pressure. The residue was purified with a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=3:1 (volume ratio)) to obtain Compound 4A (2.34 g, 80.1% yield).

MS (ESI): m/z 397,399 [M+1] ⁺ .

The second step: synthesis of compound 4B

Compound 4A (2.4g, 6.04mmol) was dissolved in dioxane (20mL), diphenylketoneimine (2g, 12.08mmol) was added, Pd ₂ (dba) ₃ (0.24g, 604μmol), Xantphos (0.35g, 1.2mmol) and Cs ₂ CO ₃ (3.9g, 12.08mmol) were stirred and refluxed at 95°C overnight under nitrogen protection. TLC showed that after the reaction was completed, it was added to saturated NH ₄ Cl aqueous solution (50 mL), and the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×10 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, filtered to remove the desiccant, and precipitated under reduced pressure. The residue was purified with a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=3:1 (volume ratio)) to obtain Compound 4B (2.74 g, 91% yield).

MS (ESI): m/z 498 [M+1] ⁺ .

The third step: synthesis of compound 4C

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, room temperature overnight. TLC showed that after the reaction was completed, the mixture was spin-dried, slurried with petroleum ether/ethyl acetate (1:1), and dried to obtain compound 4C (1.43 g, yield 86%).

MS (ESI): m/z 334 [M+1] ⁺ .

Step 4: Synthesis of Compound 4D

Compound 4C (1.0g, 3.0mmol) was added to a stuffy jar containing acetonitrile (10mL), hydrochloric acid ethyl acetate solution (5ml, mass fraction 8%) was added, and stirred overnight at 100°C. TLC showed that after the reaction was completed, a solid was precipitated after cooling to room temperature. The solid was filtered, washed with cold ethyl acetate, and dried to obtain compound 4D (0.67 g, yield 65%).

MS (ESI): m/z 343 [M+1] ⁺ .

Step 5: Synthesis of compound 4F

DMF (5mL) was added in the flask, compound 4D (0.45g, 1.31mmol) was added in the flask, and compound 4E (0.31g, 1.31mmol), DBU (0.4g, 2.62mmol) and BOP (0.87g, 1.97mmol) were added mmol), stirred at 60°C for 18h. TLC showed that after the reaction was completed, it was diluted with ethyl acetate and added to saturated NH ₄ Cl aqueous solution (50 mL), the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×10 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, filtered to remove the desiccant, and precipitated under reduced pressure. The residue was purified with a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=3:1 (volume ratio)) to obtain Compound 4F (263 mg, light yellow solid, yield 36%).

MS (ESI): m/z 559 [M+1] ⁺ .

Step 6: Synthesis of compound 4

Compound 4F (178 mg, 0.32 mmol) was added into methanol (5 mL) at room temperature, and then 10% palladium on carbon (20 mg) was added. After the addition, put a hydrogen balloon on the mouth of the bottle, and after replacing the gas three times, keep the hydrogen atmosphere at room temperature for 3 hours. After TLC showed that the reaction was over, it was filtered, the solid was washed with methanol, the filtrate was collected, the solvent was evaporated under reduced pressure, and the residue was purified on a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate=5:1 (volume ratio)) to obtain Compound 4 (108 mg, gray solid, 64% yield).

MS (ESI): m/z 529 [M+1] ⁺ .

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).

Embodiment 5: the preparation of compound 5

synthetic route:

Preparation:

The first step: synthesis of compound 5

DMF (5mL) was added to the flask, compound 4D (0.45g, 1.31mmol) was added to the flask, and compound 5A (0.25g, 1.31mmol), DBU (0.4g, 2.62mmol) and BOP (0.87g , 1.97mmol), stirred at 60°C for 18h. TLC showed that after the reaction was completed, it was added to saturated NH ₄ Cl aqueous solution (50 mL), and the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (3×10 mL). After the organic phases were combined, they were dried with anhydrous sodium sulfate, filtered to remove the desiccant, precipitated under reduced pressure, and the residue was purified with a preparative silica gel plate (developing solvent: petroleum ether/ethyl acetate = 1:1 (volume ratio)) to obtain Compound 5 (263 mg, pale yellow solid, yield 36%).

MS (ESI): m/z 514 [M+1] ⁺ .

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), 5.90-5.78(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 = 7.1 Hz, 3H).

Embodiment 6: the preparation of compound 6-1 and 6-2

synthetic route:

Preparation:

Step 1: Synthesis of a mixture of compounds 6C-1 and 6C-2

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

MS (ESI): m/z 267 [M+1] ⁺ .

Step 2: Synthesis of a mixture of compounds 6D-1 and 6D-2

The mixture of compounds 6C-1 and 6C-2 (25g, 93.6mmol) was added in portions to concentrated sulfuric acid (250mL) under ice-cooling, and while the ice-bath was maintained, potassium nitrate (9.4g, 93.6mmol) was slowly added in portions, Keep the internal temperature below 5°C, remove the ice bath after the addition, and continue the reaction for 2 hours after warming up to room temperature. TLC showed that after the reaction was completed, the reaction solution was slowly added to the stirred ice water, keeping the water temperature not exceeding 10°C, a large amount of solids precipitated after the addition, filtered, the solids were washed with water several times, and dried to obtain compounds 6D-1 and 6D-2 (26 g, white solid, 89% yield).

MS (ESI): m/z 312 [M+1] ⁺ .

Step 3: Synthesis of a mixture of compounds 6E-1 and 6E-2

The mixture (25g, 80mmol) of compound 6D-1 and 6D-2 was dissolved in the mixed solvent of ethanol (250mL) and water (50mL), and iron powder (22g, 400mmol) and ammonium chloride (21g, 400mmol) were added at room temperature ), and the reaction solution was heated to reflux overnight. After TLC showed that the reaction was over, the reaction solution was filtered and spin-dried, and the residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1:1 (volume ratio)) to obtain compounds 6E-1 and 6E -2 mixture (19.8 g, yellow solid, 88% yield).

MS (ESI): m/z 282 [M+1] ⁺ .

Step 4: Synthesis of a mixture of compounds 6F-1 and 6F-2

In a 250 mL stuffy jar, the mixture of compounds 6E-1 and 6E-2 (14.1 g, 50 mmol) was dissolved in 4N hydrochloric acid acetonitrile solution (100 mL), sealed, heated to 110° C., and reacted for 8 h. TLC showed that after the reaction was completed, a large amount of solids precipitated after cooling to room temperature, filtered, and the solids were washed with ether and dried to obtain a mixture of compounds 6F-1 and 6F-2 (11.9 g, white solid, yield 82%).

MS (ESI): m/z 291 [M+1] ⁺ .

Step 5: Synthesis of compounds 6-1 and 6-2

DMF was added in the flask, the mixture of compound 6F-1 and 6F-2 (0.38g, 1.31mmol) was added in the flask, compound 5A (0.25g, 1.31mmol), DBU (0.4g, 2.62mmol) and BOP were added (0.87g, 1.97mmol), stirred at 60°C for 18h. TLC showed that after the reaction was completed, diluted with ethyl acetate, extracted with water, and dried to obtain a mixture of compounds 6F-1 and 6F-2, which was then separated by prep-TLC (developing solvent: petroleum ether/ethyl acetate=3:1 (volume ratio)) to obtain the large polar component (25mg) and the small polar component compound (22mg) respectively.

Small polar components:

MS (ESI): m/z 462 [M+1] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ) 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

Large polar components:

MS (ESI): m/z 462 [M+1] ⁺ .

¹ H-NMR (400MHz, CDCl ₃ ): 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).

Verification of activity

Experimental example 1: KRAS (G12C) and SOS1 binding experiment

This assay can be used to examine the potency of compounds to inhibit the protein-protein interaction between SOS1 and KRAS G12C. Lower _IC50 values indicate high potency of compounds as SOS1 inhibitors in the following assay settings.

1. Experimental materials:

KRAS (G12C) protein was synthesized by Pujian Biotechnology Co., Ltd.;

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

Anti-XL665 monoclonal antibody labeled with 6 histidine tag (Mab Anti 6HIS-XL665) and anti-europium cryptate monoclonal antibody labeled with glutathione thiol transferase tag (Mab Anti GST-Eu cryptate) were purchased from Cisbio.

2. Experimental method:

1X buffer preparation (ready to use): Hepes: 5mM; NaCl: 150mM; EDTA: 10mM; Igepal: 0.0025%; KF: 100mM; DTT: 1mM;

The compound to be tested was diluted 3 times to the 8th concentration, that is, from 100 μM to 45.7 nM.

Each gradient dilution of the compound to be tested was made into a 2% DMSO working solution with 1X buffer solution, and 5 μL/well was added to the corresponding well to set up a double-well experiment. Under the condition of 1000rpm, centrifuge for 1min.

A mixed working solution of KRAS(G12C) (200nM) and Mab Anti GST-Eu cryptate (1ng/μL) was prepared with 1X buffer, and the mixed working solution was incubated at 25°C for 5min, and 2.5μL/well was added to the corresponding well.

Prepare a mixed working solution of SOS1 (80nM) and Mab Anti 6HIS-XL665 (8g/μL) with 1X buffer, add 2.5μL/well to the corresponding well, add 2.5μL Mab Anti 6HIS-XL665 (8g/μL) to the Blank well ) diluent, at this time the final concentration of the compound was diluted to 0.457nM with a gradient of 1μM, KRAS (G12C) (500nM), MAb Anti GST-Eu cryptate (0.25ng/μL), SOS1 (20nM), Mab Anti 6HIS-XL665 (2g /μL), and the reaction system was placed at 25°C for 60 minutes. After the reaction, HTRF was read using a multi-label analyzer.

3. Data analysis:

Using the equation (sample-Min)/(Max-Min)×100%, the raw data is converted into an inhibition rate, and the IC ₅₀ value can be obtained by curve fitting with four parameters (log(inhibitor)vs.response in GraphPad Prism --Variable slope mode derived). Table 1 provides the inhibitory activity of compounds of the present invention on KRAS(G12C) and SOS1 binding.

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

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

Table 1. _IC50 data of the compounds of the present invention on KRAS (G12C) and SOS1 binding inhibition

Compound number IC₅₀(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 Large Polar Components 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

It can be seen from Table 1 that the compound of the present invention has a good inhibitory effect on SOS1, and has an obvious inhibitory effect on the combination of KRAS (G12C) and SOS1, and has a good clinical application prospect.

Experimental Example 2: Cell Proliferation Inhibition Experiment

The cell proliferation inhibition assay is used to examine the efficacy of compounds in inhibiting SOS1-mediated proliferation, growth and apoptosis of cancer cell lines in vitro. Lower _IC50 values indicate high potency of compounds as SOS1 inhibitors in the following assay settings. In particular, it was observed that compounds that are SOS1 inhibitors exhibit potent inhibitory effects on the proliferation of KRAS mutant human cancer cell lines, whereas KRAS wild-type human cancer cell lines do not. This confirms the molecular mode of action of compounds that act as SOS1 inhibitors to selectively target cancer cells that depend on the function of RAS family proteins.

1. Experimental materials:

RPMI1640 medium, fetal bovine serum, containing penicillin/streptomycin double antibodies were purchased from Vicente;

Low melting point agarose was purchased from Sigma;

Almar blue reagent was purchased from Invitrogen;

NCI-H358 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd.;

Nivo Multilabel Analyzer was purchased from Perkin Elmer.

2. Experimental method:

Plant H358 cells in a 96-well U-shaped plate, first make a 2% mother solution of low-melting point agarose, heat the agarose mother solution in a microwave oven to completely melt it, and then place it in a water bath at 42°C , to keep the agarose in a liquid state. The gel was added to the serum-containing medium to make a gel concentration of 0.6% as the bottom layer gel, and spread in a 96-well U-shaped plate at a rate of 50 μL/well. After the bottom layer gel is solidified, add 2% gel to the cell-containing medium to prepare a cell-containing upper layer gel with a gel concentration of 0.4%, and the cell density is ^4x104 cells/mL. Into a 96-well U-plate covered with bottom gel, the cell density is 3000 cells/well. After the upper gel was solidified, the cell plate was placed in a carbon dioxide incubator for overnight culture.

On the day of adding the compound, 85 μL of liquid medium was added to the 96-well U-shaped plate on which the cells were laid. The compound to be tested was diluted 3 times to the 9th concentration with a row gun, that is, diluted from 6mM to 0.9μM, and a double-well experiment was set up. Add 97 μL of medium to the middle plate, and then transfer 2.5 μL/well of the gradient dilution compound to the middle plate according to the corresponding position. After mixing, transfer 40 μL/well to the cell plate. Compound concentrations ranged from 30 [mu]M to 4.5 nM transferred to the cell plate. The cell plate was cultured in a carbon dioxide incubator for 7 days. On the 8th day, the compound to be tested was diluted 3 times to the ninth concentration, that is, diluted from 6mM to 0.9μM, and a double-well experiment was set up. Add 198 μL medium to the middle plate, and then transfer 2 μL/well of the gradient dilution compound to the first middle plate according to the corresponding position, then add 100 μL medium to the second middle plate, and take the first middle plate The mixed compound (100 μL) was added, and after mixing, 40 μL/well was transferred to the cell plate. Compound concentrations ranged from 30 [mu]M to 4.5 nM transferred to the cell plate. Place the cell plate in a carbon dioxide incubator for another 7 days. The compound was co-incubated with the cells for 14 days, 20 μL/well of Almar blue detection reagent was added to the cell plate, the plate with the dye was placed on a horizontal shaker for 15 min, and the plate was incubated at room temperature for 5 h to stabilize the luminescent signal. Read using a multi-label analyzer.

3. Data analysis:

Using the equation (sample-Min)/(Max-Min)*100%, the original data is converted into an inhibition rate, and the IC ₅₀ value can be obtained by curve fitting with four parameters ("log(inhibitor) vs. response--Variable slope" mode). Table 2 provides the inhibitory activity of the compounds of the present invention on the proliferation of NCI-H358 cells. where ND means not tested.

Table 2. IC ₅₀ data of compounds of the present invention on NCI-H358 cell proliferation inhibition

Compound number IC₅₀(nM)/NCI-H358 Reference compound BI-3406 50.76 Compound 1 585 Compound 2 ND Compound 3 ND Compound 4 ND Compound 5 ND Large Polar Components 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

It can be seen from Table 2 that the compound of the present invention has a good anti-proliferation effect on human non-small cell lung cancer NCI-H358, and the cell activity is less than 1 μM, which has a good clinical application prospect.

Experimental example 3: p-ERK experiment

1. Experimental materials:

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

2. Experimental method:

DLD-1 cells were planted in a transparent 96-well cell culture plate, 80 μL of cell suspension per well, each well contained 8000 DLD-1 cells, the cell plate was placed in a carbon dioxide incubator, and incubated overnight at 37;

The compound to be tested was diluted to 2mM with 100% DMSO as the first concentration, and then diluted 5 times to the eighth concentration with a pipette, that is, diluted from 2mM to 0.026μM. Take 2 μL of the compound and add 78 μL of cell starvation medium, mix well, take 20 μL of the compound solution and add it to the corresponding well of the cell plate, put the cell plate back into the carbon dioxide incubator and continue to incubate for 1 hour. 0.5%;

After the incubation, discard the cell supernatant and add 50 μL of cell lysate to each well, shake and incubate at room temperature for 30 minutes;

Use Detection buffer to dilute Phospho-ERK1/2Eu Cryptate antibody and Phospho-ERK1/2d2antibody 20 times;

Take 16 μL cell lysate supernatant per well into a new 384 white microwell plate, then add 2 μL Phospho-ERK1/2Eu Cryptate antibody dilution and 2 μL Phospho-ERK1/2d2 antibody dilution, and incubate at room temperature for 4 hours;

After incubation, use a multi-label analyzer to read HTRF excitation: 320nm, emission: 615nm, 665nm.

3. Data analysis:

Use the equation (Sample-Min)/(Max-Min)*100% to convert the original data into an inhibition rate, and the value of IC50 can be obtained by curve fitting with four parameters (log(inhibitor)vs.response- in GraphPad Prism) -Variable slope mode derived). where ND means not tested.

Table 3. IC ₅₀ data of compounds of the present invention inhibiting phosphorylation of DLD-1 cells

Compound number IC₅₀(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 Large Polar Components 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

It can be seen from Table 3 that the compounds of the present invention, such as compounds 4 and 5, have a good inhibitory effect on the phosphorylation of DLD-1 cells, and have good clinical application prospects.

Although the embodiments of the present invention have been shown and described above, it should be understood that the above embodiments are exemplary and should not be construed as limitations on the present invention. Without departing from the principles and purposes of the present invention, those skilled in the art can change, modify, replace and modify the above-mentioned embodiments within the scope of the present invention, and these changes, modifications, replace and modify are all covered in within the scope of the present invention.

Claims (13)

1. A compound as shown in formula I or pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof,

wherein,

a is selected from C₆-C₁₀Aryl, 5-to 6-membered monocyclic heteroaryl, and 9-to 10-membered bicyclic heteroaryl, each optionally substituted with up to 5R₆Substitution;

x and Y are each independently selected from CR₆And N;

Q₁and Q₂Each independently selected from-O-, -C (R)₉)₂-and-NR₉-；

L₁And L₂Each independently selected from- (CH)₂)_m-or- (CH)₂)_m-O-(CH₂)_p-O-(CH₂)_n-wherein each of m, n and p is independently any integer from 0 to 8;

R₁and R₂Each independently selected from hydrogen and C₁-C₈An alkyl group; or R₁And R₂Together with the carbon atom to which they are attached form C₃-C₆A cycloalkylene group;

R₃selected from hydrogen, halogen, cyano, hydroxy, amino, -NH (R)₆)、-C(＝O)-NH(R₆)、C₁-C₆Alkyl radical, C₂-C₄Alkenyl radical, C₂-C₄Alkynyl, C₃-C₆Cycloalkyl, 3-to 8-membered heterocycloalkyl, C₁-C₃Alkoxy and C₁-C₆Haloalkyl, said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkoxy, and haloalkyl each optionally substituted with at least 1R₆Substitution;

R₄selected from hydrogen, halogen, cyano, hydroxy, amino, -NH (R)₆)、-C(＝O)-NH(R₆)、C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl, 3-to 8-membered heterocycloalkyl, C₁-C₃Alkoxy and C₁-C₆Haloalkyl, said alkyl, cycloalkyl, heterocycloalkyl, alkoxy, and haloalkyl each optionally substituted with at least 1R₆Substitution;

R₅selected from hydrogen, halogen, cyano, hydroxy, amino, -N (R)₆)(R₇)、-C(＝O)-N(R₆)(R₇)、-C(＝O)-R₇、-C(＝O)-OR₇、C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl, 3-to 8-membered heterocycloalkyl, C₁-C₃Alkoxy and C₁-C₆Haloalkyl, said alkyl, cycloalkyl, heterocycloalkyl, alkoxy, and haloalkyl each optionally substituted with at least 1R₆Substitution;

if present, each R₆And R₇Each independently selected from hydrogen, halogen, cyano, hydroxy, amino, carbamoyl, C₁-C₆Alkyl radical, C₁-C₆Heteroalkyl group, C₃-C₈Cycloalkyl, 3-to 14-membered heterocycloalkyl, C₁-C₃Alkoxy radical, C₁-C₃Haloalkoxy, C₆-C₁₀Aryl, 5-to 6-membered monocyclic heteroaryl and 9-to 10-membered bicyclic heteroaryl, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, haloalkoxy, aryl, monocyclic heteroarylEach of which is optionally substituted with at least 1R₈Substitution; or R₆And R₇Taken together with the nitrogen atom to which they are attached form a 5-to 6-membered heterocycloalkyl optionally substituted with at least 1R₈Substitution;

if present, each R₈Each independently selected from hydrogen, chloro, fluoro, cyano, hydroxy, amino, isopropyl, cyclopropyl, methyl, difluoromethyl, trifluoromethyl, methoxy, trifluoromethoxy, ethoxy, 2-difluoroethoxy, 2-trifluoroethoxy, and phenyl;

if present, each R₉Each independently selected from hydrogen, halogen, cyano, hydroxy, amino, -N (R)₆)(R₇)、-C(＝O)-N(R₆)(R₇)、-C(＝O)-R₇、-C(＝O)-OR₇、C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl, 3-to 8-membered heterocycloalkyl, C₁-C₃Alkoxy and C₁-C₆Haloalkyl, said alkyl, cycloalkyl, heterocycloalkyl, alkoxy, and haloalkyl each optionally substituted with at least 1R₆And (4) substitution.

2. The compound of claim 1, wherein the compound is represented by formula I-1,

wherein,

q is any integer of 0 to 4;

X、Y、R₁、R₂、R₃、R₄、R₅and R₆As defined in claim 1.

3. The compound of claim 1 or 2, wherein the compound is of formula I-1-1 or formula I-1-2,

wherein,

q is any integer of 0 to 4;

X、Y、R₁、R₂、R₃、R₄、R₆and R₇As defined in claim 1.

4. A compound according to claim 3,

the group is selected from any one of the following groups:

5. the compound of claim 1, wherein the compound is represented by formula I-2,

wherein,

q is any integer of 0 to 4;

X、Y、R₁、R₂、R₃、R₄、R₅and R₆As defined in claim 1.

6. The compound of claim 1 or 5, wherein the compound is a compound of formula I-2-1,

wherein,

q is any integer of 0 to 4;

X、Y、R₁、R₂、R₃、R₄and R₆As defined in claim 1.

7. The compound of claim 1, wherein the compound is represented by formula I-3,

wherein,

q is any integer of 0 to 4;

X、Y、R₁、R₂、R₃、R₄、R₅、R₆and R₉As defined in claim 1.

8. The compound of claim 1 or 7, wherein the compound is a compound of formula I-3-1,

wherein,

q is any integer of 0 to 4;

X、Y、R₁、R₂、R₃、R₄、R₆and R₉As defined in claim 1.

9. A compound according to claim 8,

R₉any one selected from the following groups:

10. a pharmaceutical composition comprising a compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite, or prodrug thereof, and at least one pharmaceutically acceptable adjuvant.

11. A compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, or a pharmaceutical composition according to claim 10, for use as a SOS1 inhibitor or for use in the prevention and/or treatment of a disease or condition caused by the overexpression of SOS 1;

preferably, the disease or disorder caused by overexpression of SOS1 is cancer, preferably pancreatic cancer, colorectal cancer and lung cancer.

12. Use of a compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, or a pharmaceutical composition according to claim 10, for the manufacture of a medicament for the prevention and/or treatment of a disease or condition caused by overexpression of SOS 1;

preferably, the disease or disorder caused by overexpression of SOS1 is cancer, preferably pancreatic cancer, colorectal cancer and lung cancer.

13. A method for preventing and/or treating a disease or disorder caused by overexpression of SOS1, comprising administering to a subject in need thereof a prophylactically and/or therapeutically effective amount of a compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, or a pharmaceutical composition according to claim 10;

preferably, the disease or disorder caused by overexpression of SOS1 is cancer, preferably pancreatic cancer, colorectal cancer and lung cancer.