Preparation and application of SOS1 protein inhibitor

Abstract

The invention belongs to the technical field of medical chemistry, particularly relates to a compound capable of being combined with SOS1 and a composition of the compound, and further relates to application of the compound capable of being combined with SOS1 and the composition of the compound. Specifically disclosed are a compound capable of binding SOS1, and pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers thereof, the compound being as shown in (I); the synthesized compound can be effectively combined with SOS1 protein, so that activation of SOS1 on downstream protein abnormal KRAS is inhibited, cancer cell proliferation caused by abnormal KRAS activation is inhibited, the effect of inhibiting cancer cell proliferation is finally achieved, and IC50 of the compound can reach 10 mu M or below. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a compound capable of binding to SOS1 and its composition, and also relates to the use of the compound capable of binding to SOS1 and its composition. Background Art

[0002] The first human proto-oncogene RAS was discovered by Weinberg and Barbacid in 1982. Subsequent studies have confirmed that cell malignant transformation is related to gene mutations encoding the RAS protein. The smooth protein structure of RAS and its high affinity for guanosine triphosphate (GTP) have made RAS once known as an "undruggable target". SOS1 (Son of sevenless 1), as an upstream protein of RAS, is related to the activation of RAS. As a molecular switch of the downstream pathway, the RAS protein relies on guanine nucleotide exchange factor (GEF) to cycle between the inactive state (RAS-GDP) and the active state (RAS-GTP). The RAS protein itself has certain GTPase activity, and GTPase-activating protein (GAP) helps it promote the hydrolysis of GTP. The GAP activity of the mutant RAS protein is significantly reduced, resulting in permanent activation of RAS. The activation of the RAS pathway is regulated by signaling molecules. Signaling molecules such as epidermal growth factor (EGF) bind to tyrosine kinase receptors, resulting in phosphorylation of the latter receptor. The activated tyrosine kinase receptor binds to the SH2 domain of growth factor receptor-bound protein 2 (Grb2). At the same time, the SH3 domain of Grb2 recruits SOS protein to form a complex, localizes the SOS protein to the cytoplasmic membrane, and the SOS protein localized on the membrane catalyzes the binding of RAS to GTP. RAS-GTP can activate downstream pathways, among which the most important are the mitogen-activated protein kinase (MAPK) pathway and the phosphatidylinositol 3-kinase (PI3K) / protein kinase B (Akt) / mammalian target of rapamycin (mTOR) pathway. Among them, the protein encoded by the KRAS gene is a small GTPase, which belongs to the RAS superprotein family; in normal cells, receptor monomers such as EGFR, HER2, ErbB3, and ErbB4 on the cell membrane bind to extracellular ligands to form dimers, and the dimers are phosphorylated by themselves and then phosphorylate downstream signaling proteins; one of the signaling pathways can activate Grb2-Shc, then activate the SOS protein, and further activate the KRAS protein. It has been found that reducing the expression of SOS1 or inhibiting its activity can reduce the proliferation rate and survival rate of KRAS mutant cells. Therefore, inhibitors targeting SOS1 can not only be used to treat tumors with KRAS mutations, but also overcome the drug resistance caused by KRAS inhibitors.

[0003] The SOS1 protein has multiple domains, including HD, DH, PH, HL, REM, CDC25H, and PR. Among them, REM is an allosteric site regulated by RAS-GTP. CDC25H is a catalytic site that promotes the conversion of GDP to GTP by adjusting the direction of its own helical hairpin. REM, CDC25H, and PR together participate in the formation of RAS-GTP. PH is related to the membrane localization of SOS1. In the resting state, DH, PH, and PR form an autoinhibitory module that can block the binding site of the REM domain and inhibit the catalytic activity of the CDC25H domain, thereby inhibiting the physiological function of SOS1; when the REM domain of SOS1 is activated by RASGTP, its autoinhibitory state is relieved and the GEF function is restored, which is the activation mechanism of SOS1 regulated by positive feedback. Studies have found that mutations at the SOS1 W729 site can disrupt the binding of the allosteric site to RAS; mutations at the SOS1 F929 site can disrupt the catalytic activity of the CDC25H domain and affect the membrane localization of SOS1. The membrane localization of SOS1 plays an important role in the exertion of its biological function. The Grb2 protein that helps localize it has two domains - nSH3 and cSH3. These two domains have high affinity for the PVPPPVPPRRRP and PKLPPKTYKREH polypeptides in the PR domain of the SOS1 protein, respectively. Disrupting the above polypeptides will seriously affect the biological activity of SOS1. The SOS1 S1178 site is also a key site for SOS1 membrane localization. Phosphorylation at this site can block the binding of Grb2 to SOS1, disrupting the membrane localization of SOS1.

[0004] The SOS1 protein has received extensive attention in RAS-related diseases (such as Noonan syndrome). Recently, some scholars have also found that SOS1 is involved in the development of different tumors. For example, in lung adenocarcinoma, mutant SOS1 can not only up-regulate the expression of RAS-related genes, activate the MAPK pathway, but also up-regulate the expression of c-MYC, promoting the proliferation of tumor cells. In gastric cancer, SOS1 is trans-activated by RUNX-related transcription factor 1 (RUNX1), enhancing the activation of the ErbB2 / HER2 signaling pathway and promoting the malignant development of gastric cancer cells. In ovarian cancer, the SOS1 protein is mainly related to the invasion and metastasis of tumor cells. The SOS1 protein forms a complex with ABI1 and EPS8, and this complex is the key to the metastasis of ovarian cancer cells. In glioblastoma, the high expression of the SOS1 protein can not only reduce the sensitivity of T98G cells to the combined use of miR-152-3p (a kind of miRNA) and cisplatin, but also be related to the desensitization of miR-152-3p in the treatment of glioblastoma. To sum up, the tumor-promoting effect of the SOS1 protein is not only related to its activation of MAPK and its downstream pathways, but also related to the high expression of its own protein level. Therefore, drugs targeting SOS1 can not only inhibit the activation of RAS protein, but also inhibit the function of SOS1 protein, thus inhibiting the malignant process of tumor cells.

[0005] Currently, the small molecule inhibitor targeting SOS1 is NSC-658497, which is a lead inhibitor that can block the interaction between SOS1 and RAS. It inhibits the activity of SOS1 GEF in a dose-dependent manner at the micromolar level, but the research results of it in animal experiments have not been reported yet. BAY-293 is a quinazoline inhibitor obtained by Bayer through high-throughput fragment screening. It can disrupt the binding of KRAS G12C to the SOS1 protein (IC50 = 21 nmol·L-1). Currently, no clinical research related to this compound has been found. BI-3406 is developed by Boehringer Ingelheim and is an orally effective and selective SOS1 inhibitor. BI-3406 can block the protein-protein interaction between SOS1 and KRAS. Moreover, BI-3406 can inhibit p-ERK1 / 2 in KRAS G12 / G13 / Q61 mutant cells or wild-type cells (IC 50It is 17 - 57 nmol·L-1). Compared with the Q61 site mutation, the G12 and G13 sites are more sensitive to BI-3406. However, BI-3406 cannot inhibit the proliferation of wild-type KRAS cells; meanwhile, some cells with mutated KRAS and also mutated BRAF (such as HT-29 cells, A375 cells) are not sensitive to the treatment of BI-3406. Therefore, the effect of monotherapy with BI-3406 is limited and it needs to be combined with other inhibitors; BI-1701963 is the first SOS1 inhibitor to enter phase I clinical research. By binding to the catalytic region of SOS1, BI-1701963 can inhibit the binding of SOS1 to KRAS-GDP, reduce the formation of KRASGTP, and thus inhibit the activation of the MAPK signaling pathway. Even for BI-1701963 that has entered clinical trials, the publicly disclosed data are scarce, and its efficacy and potential toxicity are not yet clear. Continuing to search for SOS1 inhibitors with high target specificity and low toxicity is the focus of subsequent research on this class of drugs. SUMMARY OF THE INVENTION

[0006] The present invention provides a compound capable of binding to SOS1 protein and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers, and the compound is as shown in (I):

[0007]

[0008] R1 is selected from one or more of H, D (deuterium), F, Cl, Br, I, OH, OR, CN, NH2, NHR, NRR’, COOH, COOR, CONH2, CONHR, CONRR’, R, CF3, CHF2 or CH2F, wherein R or R’ is each independently selected from C1 - 6 alkyl;

[0009] W is selected from one of O and C,

[0010] When W is O, R2 and R3 are absent;

[0011] When W is C, R2 and R3 may not be connected to each other, in which case R2 = R3, and R2, R3 are selected from CH3 or F;

[0012] When W is C, R2 and R3 may be connected to each other, in which case R2 = R3 = -(CH2)-, and W, R2 and R3 form a cyclopropyl group;

[0013] R4 is selected from one or more of the following:

[0014]

[0015] Ar is an aromatic ring or an aromatic heterocycle, and the Ar structure is selected from one or more of the following:

[0016]

[0017]

[0018] Among them, X in the Ar structure is selected from one or more of O, S, SO2, NH, and NMe;

[0019] The specific substituents R5, R6, R7, R8, and R9 in the above Ar structure are all selected from: H, D (deuterium), F, Cl, Br, I, C 1～6 alkyl, C 1～6 mono- or polyhaloalkyl, C 1～6 mono- or polydeuterated alkyl, C 3～6 cycloalkyl, C 3～6 mono- or polyhalocycloalkyl, C 3～6 mono- or polydeuterated cycloalkyl, CN, NO2, NH2, carbonyl oxygen, OH, or one or more of compounds with other structures; the compounds with other structures are shown as follows:

[0020]

[0021] Among them, (R m ) n represents the substituents and their numbers connected to Ar, and R m represents the specific substituents of the Ar structure, and R m is selected from one or more of R5, R6, R7, R8, or R9; n is the number of R m , and n is an integer taken from 1 to 5.

[0022] Furthermore, the pharmaceutically acceptable salt refers to a salt formed by adding a non-toxic acid or its base to the parent compound.

[0023] Furthermore, the alkyl includes but is not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl, and sec-pentyl.

[0024] In a second aspect, the present invention provides a pharmaceutical composition, which comprises a combination of the compound of formula (I) described in the first aspect, its pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer, and a pharmaceutically acceptable excipient or carrier.

[0025] In a third aspect, the present invention provides an inhibitor for treating or preventing diseases caused by abnormal activation of KRAS.

[0026] Furthermore, the inhibitor comprises the compound (I) described in the first aspect of the present invention or its pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer.

[0027] Furthermore, the inhibitor can bind to the SOS1 protein, thereby inhibiting the abnormal activation of the downstream protein KRAS by the SOS1 protein.

[0028] Furthermore, the disease refers to a tumor, and the tumor includes but is not limited to myeloid leukemia.

[0029] In a fourth aspect, the present invention provides a preparation for inhibiting the proliferation of tumor cells, and the preparation contains the compound capable of binding to the SOS1 protein described in the first aspect and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers.

[0030] In a fifth aspect, the present invention provides the use of the compound capable of binding to the SOS1 protein described in the first aspect and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers in the preparation of a preparation for inhibiting the proliferation of tumor cells.

[0031] In a sixth aspect, the present invention provides the use of the compound capable of binding to the SOS1 protein described in the first aspect and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers in the preparation of a preparation for regulating a disease caused by the abnormal activation of KRAS.

[0032] Beneficial effects

[0033] The compound synthesized by the present invention can effectively bind to the SOS1 protein, thereby inhibiting the abnormal activation of SOS1 on the downstream protein KRAS, thereby inhibiting the proliferation of cancer cells caused by the abnormal activation of KRAS, and finally playing a role in inhibiting the proliferation of cancer cells, and its IC 50 can reach below 10 μM. Description of the drawings

[0034] Figure 1 . Proliferation inhibition curve of series A compounds against K562 cell line.

[0035] Figure 2 . Proliferation inhibition curve of series B compounds against K562 cell line.

[0036] Figure 3 . Proliferation inhibition curve of series C compounds against K562 cell line.

[0037] Figure 4 . Proliferation inhibition curve of series D compounds against K562 cell line.

[0038] Figure 5 . Proliferation inhibition curve of series S compounds against K562 cell line. Detailed implementation manners

[0039] The following further describes the specific embodiments of the present invention. It should be noted here that the description of these embodiments is for helping to understand the present invention, but does not constitute a limitation to the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0040] The experimental methods in the following examples are all conventional methods unless otherwise specified. The test materials used in the following examples are all commercially available through conventional commercial channels unless otherwise specified.

[0041] As used herein, the term "alkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups and has a specified number of carbon atoms, generally from 1 to about 12 carbon atoms. As used herein, the term C1-C6 alkyl represents an alkyl group having from 1 to about 6 carbon atoms. Examples of alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl, and sec-pentyl.

[0042] As used herein, the term "stereoisomer" refers to compounds having the same chemical composition but different arrangements of atoms or groups in space, which includes "diastereomers" and "enantiomers".

[0043] As used herein, the term "diastereomer" refers to stereoisomers having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as: melting point, boiling point, spectral characteristics, and reactivity. In the presence of a resolving agent or chromatography, a mixture of diastereomers can be separated under high-resolution analysis steps such as electrophoresis and crystallization using, for example, a chiral HPLC column.

[0044] As used herein, the term "enantiomer" refers to two stereoisomers of a compound that are non-overlapping mirror images of each other. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur in chemical reactions or processing when there is no longer stereoselectivity or stereospecificity.

[0045] As used herein, the terms "pharmaceutically acceptable salts" and "salts of the compounds" are interchangeable and are derivatives of the disclosed compounds, wherein the parent compound is prepared by forming non-toxic acid or its base addition salts; examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts: salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid. Other pharmaceutically acceptable salts include, but are not limited to, adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, fumarates, glucoheptanoates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactates, lactobionates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide ions, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having 1 to 6 carbon atoms, sulfonate, and arylsulfonate.

[0046] Examples

[0047] In nuclear magnetic resonance spectroscopy (NMR), 1H (400 MHz) was measured using a JEOL JNM-EPC 400 NMR nuclear magnetic resonance spectrometer, and 1H (500 MHz) and 13C-NMR (125 MHz) were measured using an Agilent Pro Pulse 500 NMR nuclear magnetic resonance tester. DMSO-d6 (TMS as the internal standard) was used as the solvent. The nuclear magnetic resonance spectra were expressed in ppm for chemical shift (δ), and the peak patterns were represented by singlet (s), doublet (d), doublet of doublets (dd), triplet (t), quartet (q), multiplet (m), etc., as well as several peak patterns combined with the above splitting types; ultra performance liquid chromatography-mass spectrometry (UPLC-MS) was measured using Waters ACQUITY UPLC, and the test method was Method A: 5 to 100% MeCN / H2O with 0.1% FA in 10 min.

[0048] Synthesis of Intermediates E1-A, E1-273, E1-276 to 278 in Example 1

[0049] Dissolve bromo-fluorobenzaldehyde or bromo-fluorobenzophenone (1.0 eq.) in DCM, add BAST (1.5 eq.) at 0 °C, stir at room temperature for 12 h, detect by TLC until the raw materials are completely consumed, add saturated aqueous NaHCO3 solution, extract with DCM (dichloromethane), combine the organic phases, dry with anhydrous Na2SO4, filter, and remove the solvent by rotary evaporation under reduced pressure. The crude product is purified by column chromatography to obtain intermediate E1-A or E1-273 or E1-276 or E1-277 or E1-278.

[0050]

[0051] Synthetic Route 1

[0052]

[0053] Synthetic Routes of Intermediates E5-A, E5-B, E5-C, and E5-D in Example 2

[0054]

Step1

[0055]

Step2

[0056]

Step3

[0057]

Step4

[0058]

[0059] The syntheses of intermediates E5-A, E5-B, E5-C, E5-D, and E5-271 to 286 were completed according to the above route. The products and yields of each step are summarized in the following table.

[0060]

[0061]

[0062]

[0063]

[0064] N / A indicates no compound

[0065] Synthetic route of intermediate E5-C2 in Example 3

[0066] Dissolve intermediate E5-C (700.0 mg, 2.9 mmol, 1.0 eq.), 2-hydroxymethylphenylboronic acid (482.0 mg, 3.2 mmol, 1.1 eq.), Cs2CO3 (3.8 g, 11.6 mmol, 4.0 eq.), and Pd(PPh3)4 (335 mg, 0.3 mmol, 0.1 eq.) in THF:H2O = 10:1 (11 mL). Heat to 70 °C and stir under an argon atmosphere. After TLC detection shows complete consumption of the starting material, remove the solvent by rotary evaporation under reduced pressure. Dissolve in DCM, wash the organic phase with water, combine the organic phases, dry over anhydrous Na2SO4, filter, and remove the solvent by rotary evaporation under reduced pressure. Purify the crude product by column chromatography (V DCM / V MeOH = 60 / 1 - 10 / 1) to obtain 510.0 mg of E5-C2 as a pale yellow liquid with a yield of 75%.

[0067]

[0068] Synthesis Route of Intermediate F4 in Example 4

[0069]

Step1

[0070]

Step2

[0071]

Step3

[0072]

Step4

[0073]

[0074] Synthetic route of compound F5 in Example 5

[0075] Dissolve intermediate E5 (1.0 eq.) and F4 (1.5 eq.) in anhydrous DMSO (20 mL). Add DIPEA (4.0 eq.) (N,N - diisopropylethylamine). Warm the mixture to 80 °C and stir for 12 h. After TLC detection shows complete consumption of the starting materials, cool to room temperature, add ice water, extract with EtOAc, combine the organic phases, dry over anhydrous Na2SO4, filter, and rotary evaporate the solvent under reduced pressure. Purify the crude product by column chromatography to obtain intermediate F5.

[0076]

[0077] The synthesis of intermediates F5 - A, F5 - B, F5 - C1, and F5 - D1 is completed according to synthetic route 5. The products and yields of each step are summarized in the following table.

[0078]

[0079] Synthetic route of compound F5 - C4 in Example 6

[0080]

Step1

[0081]

Step2

[0082]

Step3

[0083]

[0084] Example 7 Synthesis of Compound F5-D2

[0085] Dissolve intermediate F5-D1 (240.0 mg, 0.48 mmol, 1.0 eq.) and 10% palladium on carbon (48.0 mg, 0.09 mmol, 0.2 eq.) in MeOH (10 mL), heat and stir at 40 °C under a hydrogen atmosphere for 4 h. TLC detection showed that the raw materials were completely consumed. Filter through diatomaceous earth to remove the solid, and remove the solvent by rotary evaporation under reduced pressure. The crude product was purified by column chromatography (V DCM / V MeOH = 150 / 1 - 20 / 1), 185.0 mg of F5-D2 was obtained as a white solid with a yield of 82%.

[0086]

[0087] Example 8 Synthetic Routes of Compounds G3-A-a to s, G3-B-a to o, G3-C-a to h, G3-D-a to b

[0088]

Step 1

[0089]

Step 2

[0090]

[0091] The synthesis of intermediates G2-A-a~s, G2-B-a~o, G2-C-a~h, G2-D-a~b was completed according to the above route. The products and yields of each step are summarized in the following table.

[0092]

[0093]

[0094]

[0095]

[0096]

[0097] N / A indicates no compound

[0098] The synthesis of intermediates G3-A-a~s, G3-B-a~o, G3-C-a~h, G3-D-a~b was completed according to the above route. The products and yields of each step are summarized in the following table.

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105] N / A indicates no compound

[0106] Synthetic routes of target products A2-9a-s, B2-9a-o, and D2-10a-b in Example 9

[0107] Dissolve intermediate G3-A-a-s or G3-B-a-o or G3-D-a-b (1.0 eq.) in DCE (1,2-dichloroethane) (0.01 M), add NaBH(OAc)3 (2.0 eq.), stir the mixture at room temperature for 2 h, monitor the complete consumption of the starting material by TLC, remove the solvent by rotary evaporation under reduced pressure, and purify the crude product by pre-HPLC (method: 10%-90% MeCN, 41 min) to obtain compounds A2-9a-s or B2-9a-o or D2-10a-b.

[0108]

[0109] The synthesis of target products A2-9a-s, B2-9a-o, and D2-10a-b was completed according to the above routes, and the products and yields are summarized in the following table.

[0110]

[0111]

[0112]

[0113]

[0114]

[0115] N / A indicates no compound

[0116] Synthetic route of target product A2-9t in Example 10

[0117]

Step1

[0118]

Step2

[0119]

[0120] Example 11 Synthetic routes of target products C2-12a - h

[0121] Dissolve intermediates G3-C-a - h (1.0 eq.) in solvent DCE (0.01 M), add NaBH(OAc)3 (2.0 eq), and stir the mixture at room temperature for 2 h. Monitor the reaction by TLC until the raw materials are completely consumed. Remove the solvent by rotary evaporation under reduced pressure, dilute with EA, wash with water, combine the organic phases, dry over anhydrous Na2SO4, filter, and then remove the solvent by rotary evaporation under reduced pressure. Dissolve the obtained crude intermediate in 1,4-dioxane (0.05 M), add 4 N HCl (6.0 eq.) dioxane solution, monitor the reaction by TLC until the raw materials are completely consumed, remove the solvent by rotary evaporation under reduced pressure, and purify the crude product by pre-HPLC (method: 10% - 90% MeCN, 41 min) to obtain target products C2-12a - h.

[0122]

[0123] The synthesis of target products C2-12a - h was completed according to the above route, and the products and yields are summarized in the following table.

[0124]

[0125] Example 12 Synthetic routes of intermediates H6, H7, H8

[0126]

Step 1

[0127]

Step 2

[0128]

Step 3

[0129]

Step 4

[0130]

Step 5

[0131]

Step 6A

[0132]

Step 6B

[0133]

Step 6C

[0134]

[0135] Synthesis Route of Target Products S-271 to S-286 in Example 13

[0136] Dissolve one of the compounds in intermediate E5-271 to E5-286 (1.0 eq.) and intermediate H6 (1.3 eq.) in anhydrous DMSO (0.3 M), add DIPEA (4.0 eq.) (N,N-diisopropylethylamine), heat the mixture to 80 °C and stir for 12 h. TLC detects that the raw materials are completely consumed. Cool to room temperature, add ice water and then extract with EtOAc. The combined organic phases are dried over anhydrous Na2SO4, filtered, and the solvent is removed by rotary evaporation under reduced pressure. The crude product is purified by pre-HPLC (method: 10%-90% MeCN, 41 min) to obtain compounds S-271 to S-286.

[0137]

[0138] The synthesis of the target products S-271 to S-286 was completed according to the above route, and the structures and yields of each target product are summarized in the following table;

[0139]

[0140]

[0141] Example 14 Synthesis of Target Product S-287

[0142] Dissolve intermediate E1-A (50 mg, 0.26 mmol, 1.0 eq.) and intermediate H7 (97 mg, 0.34 mmol, 1.3 eq.) in anhydrous DMSO (0.2 M), add DIPEA (151 mg, 1.06 mmol, 4.0 eq.) (N,N-diisopropylethylamine), heat the mixture to 80 °C and stir for 12 h. Monitor the reaction by TLC until the raw materials are completely consumed. Cool the reaction mixture to room temperature, add ice water, and extract with EtOAc. Combine the organic phases, dry over anhydrous Na2SO4, filter, and concentrate the filtrate under reduced pressure to remove the solvent. Purify the crude product by pre-HPLC (method: 10%-90% MeCN, 41 min) to obtain 53 mg of compound S-287 with a yield of 43%.

[0143]

[0144] Example 15 Synthesis of Target Product S-288

[0145] Dissolve intermediate E1-A (46 mg, 0.24 mmol, 1.0 eq.) and intermediate H8 (97 mg, 0.32 mmol, 1.3 eq.) in anhydrous DMSO (0.2 M), add DIPEA (139 mg, 0.97 mmol, 4.0 eq.) (N,N-diisopropylethylamine), heat the mixture to 80 °C and stir for 12 h. Monitor the reaction by TLC until the raw materials are completely consumed. Cool the reaction mixture to room temperature, add ice water, and extract with EtOAc. Combine the organic phases, dry over anhydrous Na2SO4, filter, and concentrate the filtrate under reduced pressure to remove the solvent. Purify the crude product by pre-HPLC (method: 10%-90% MeCN, 41 min) to obtain 45 mg of compound S-288 with a yield of 40%.

[0146]

[0147] Table 1 Summary of Specific Example Information of Compounds in Series A-D and Compounds in Series S

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178] Example 16 Proliferation Inhibitory Activity of SOS1 Inhibitor on K562 Cells

[0179] The human chronic myeloid leukemia cell line K562 cells sensitive to SOS1 were selected to evaluate the proliferation inhibitory activity of four series of compounds. K562 cells (IMDM + 10% FBS + 1% PS) were seeded into a 96-well cell culture plate at 5000 cells per well, and different concentrations of the drug after gradient dilution were added. After culturing in an incubator (37 °C, 5% CO2) for 72 h, Cell-Counting-Kit-8 (CCK8) reagent with a volume of 1 / 10 of the bottom volume was added and incubated for 2 hours. The absorbance value (OD) at 450 nm was measured using a microplate reader, and the cell survival rate was calculated based on the OD value. In the proliferation inhibition data, +++ represents IC 50 < 10 μM, ++ represents IC 50 between 10 - 20 μM, + represents IC 50 > 20 μM.

[0180] 1. Proliferation Inhibition Results of Series A Compounds (A2-9a to A2-9t) on K562 Cell Line

[0181] The experimental results are as Figure 1 shown in

[0182] Table 2 IC of Series A Compounds against K562 Cell Line Inhibition 50

[0183]

[0184] 2. Proliferation Inhibition Results of Series B Compounds (B2-9a to B2-9o) on K562 Cell Line

[0185] The experimental results are as Figure 2 shown in

[0186] Table 3 IC of Series B Compounds against K562 Cell Line Inhibition 50

[0187]

[0188]

[0189] 3. Proliferation inhibition results of Series C compounds (C2-12a to C2-12h) on K562 cell line

[0190] The experimental results are as Figure 3 shown in and Table 4;

[0191] Table 4 IC of Series C compounds inhibiting K562 cell line 50

[0192]

[0193] 4. Proliferation inhibition results of Series D compounds (D2-10a and D2-10b) on K562 cell line

[0194] The experimental results are as Figure 4 shown in and Table 5;

[0195] Table 5 IC of Series D compounds inhibiting K562 cell line 50

[0196]

[0197] 5. Proliferation inhibition results of Series S compounds (S-271 to S-288) on K562 cell line

[0198] The experimental results are as Figure 5 shown in and Table 6;

[0199] Table 6 IC of Series S compounds inhibiting K562 cell line 50

[0200]

[0201]

Claims

1. A compound capable of binding to SOS1 protein, and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers, wherein the compound is shown as formula (Ⅰ): Among them, R1 is selected from one or more of H, D (deuterium), F, Cl, Br, I, OH, OR, CN, NH2, NHR, NRR’, COOH, COOR, CONH2, CONHR, CONRR’, R, CF3, CHF2 or CH2F; R or R' is each independently selected from C 1～6 alkyl; W is selected from one of O and C; When W is O, R2 and R3 are absent; When W is C, R2 and R3 may not be connected to each other, in which case R2 = R3, and R2 and R3 are selected from CH3 or F; When W is C, R2 and R3 may be connected to each other, in which case R2 = R3 = -(CH2)-, and W, R2 and R3 form a cyclopropyl group; R4 is selected from one or more of the following: Ar is an aromatic ring or a heteroaromatic ring, and the structure of Ar is selected from one or more of the following: Wherein X in the structure of Ar is selected from one or more of O, S, SO2, NH and NMe; The specific substituents R5, R6, R7, R8, and R9 in the Ar structure are each independently selected from: H, D (deuterium), F, Cl, Br, I, C 1～6 alkyl, C 1～6 mono- or polyhaloalkyl, C 1～6 mono- or polydeuteroalkyl, C 3～6 cycloalkyl, C 3～6 mono- or polyhalocycloalkyl, C 3～6 mono- or polydeutero-cycloalkyl, CN, NO2, NH2, carbonyl oxygen, OH, or one or more of compounds of other structures; the compounds of other structures are shown below: Among them, (R m ) n represents the substituents and their numbers attached to Ar, and R m represents the specific substituents of the Ar structure. R m is selected from one or more of R5, R6, R7, R8 or R9; n is the number of R m , and n is an integer taken from 1 to 5.

2. The compound capable of binding to SOS1 protein according to claim 1, and its pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer or tautomer, characterized in that, The pharmaceutically acceptable salt refers to a salt formed by the parent compound by preparing a non-toxic acid or its base addition.

3. A pharmaceutical composition, which comprises a combination of the compound capable of binding to SOS1 described in claim 1 and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers and a pharmaceutically acceptable excipient or carrier.

4. An inhibitor for treating or preventing diseases caused by abnormal activation of KRAS, characterized in that, The inhibitor comprises the compound capable of binding to SOS1 described in claim 1 and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers.

5. The inhibitor according to claim 4, characterized in that, The inhibitor can bind to SOS1 protein, thereby inhibiting the abnormal activation of downstream protein KRAS by SOS1.

6. The inhibitor according to claim 4, wherein The disease refers to a tumor, and the tumor includes but is not limited to myeloid leukemia.

7. A preparation for inhibiting the proliferation of tumor cells, which contains the compound capable of binding to SOS1 described in claim 1 and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers or the pharmaceutical composition described in claim 3.

8. Use of the compound capable of binding to SOS1 described in claim 1 and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers or the pharmaceutical composition described in claim 3 in the preparation of a preparation for inhibiting the proliferation of tumor cells.

9. Use of the compound capable of binding to SOS1 described in claim 1 and its pharmaceutically acceptable salts, hydrates, solvates, prodrugs, stereoisomers or tautomers or the pharmaceutical composition described in claim 3 in the preparation of a preparation for regulating a disease caused by the abnormal activation of KRAS.

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