Synthesis of a class of egfr inhibitors and uses thereof

Abstract

The application discloses synthesis of EGFR inhibitors and purposes thereof, and belongs to the technical field of medicines. The application relates to synthesis of a class of oxoisoindole compounds and pharmaceutically acceptable salts of the oxoisoindole compounds and purposes thereof. The compounds or the salts thereof can selectively inhibit EGFR, and thus can be used for treating or improving abnormal cell proliferation diseases.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to the synthesis and use of a class of EGFR inhibitors. Background Technology

[0002] Epidermal growth factor receptor (EGFR) is a transmembrane protein tyrosine kinase of the ErbB receptor family. When it binds to a growth factor ligand (such as epidermal growth factor (EGF)), the receptor can homodimerize with the attached EGFR molecule or heterodimerize with another family member (such as ErbB2 (HER2), ErbB3 (HER3), or ErbB4 (HER4)). Homodimerization and / or heterodimerization of the ErbB receptor leads to phosphorylation of key intracellular tyrosine residues and stimulation of many intracellular signaling pathways involved in cell proliferation and survival. Dysregulation of ErbB family signaling promotes proliferation, invasion, metastasis, angiogenesis, and tumor cell survival, and is closely related to human cancers such as lung cancer, head and neck cancer, colon cancer, and breast cancer.

[0003] Small molecule kinase inhibitors, including first-generation EGFR-TKI targeted drugs such as erlotinib and gefitinib, have shown good therapeutic effects against the gold standard mutations in EGFR, namely the deletion mutation in exon 19 (delE746-A750) and the point mutation in exon 21 (L858R). Second-generation EGFR-TKI targeted drugs, such as dacomitinib and afatinib, have not seen significant improvements in efficacy compared to first-generation drugs and have more severe side effects, thus their clinical application is not widespread. Third-generation EGFR-TKI targeted drugs, such as osimertinib and avitinib, have become the first-line treatment for EGFR-sensitive mutations and T790M-resistant mutations in NSCLC. They are adenosine triphosphate (ATP) analogs that block the activation of signaling pathways by inhibiting phosphorylation of intracellular domains.

[0004] Over the past decade, molecularly targeted therapies, exemplified by epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), have revolutionized cancer treatment. However, EGFR-TKI resistance remains a significant challenge. For instance, while EGFR-TKIs are initially effective in non-small cell lung cancer (NSCLC) patients carrying EGFR mutations, they often develop acquired resistance and lose their therapeutic efficacy. This is due to mutations such as the T790M mutation, which leads to resistance to first / second-generation EGFR-TKIs, and the C797S mutation, which leads to resistance to third-generation EGFR-TKIs. Tumors with simultaneous L858R / T790M / C797S mutations are particularly vulnerable, as current EGFR-TKIs are ineffective against them. Furthermore, all current EGFR TKIs target the ATP site, and although third-generation irreversible inhibitors (such as osimertinib) can overcome T790M, resistance arises due to the pre-existing C797S mutation in treated patients. Cetuximab (an anti-EGFR antibody) that blocks receptor dimerization is ineffective in EGFR-mutant NSCLC because kinase mutation activation is a potent "downstream" of receptor dimerization. Therefore, an alternative strategy is to inhibit EGFR. Currently, suitable compounds with alternative mechanisms of action targeting mutant EGFR are not available.

[0005] Recent studies have shown that targeted allosteric sites can lead to mutant-selective inhibitors. There is indeed a need for selective molecules that specifically inhibit EGFR mutants containing T790M / L858R, T790M / L858R / C797S, L858R, and L858R / C797S, particularly those containing T790M and C797S. These selective molecules could be used for therapeutic and / or preventative cancer treatment. Therefore, there is an urgent need for novel and effective small-molecule EGFR inhibitors with alternative mechanisms of action targeting mutant EGFR. EGFR allosteric inhibitors have been found to bind to different EGFR sites in competition with existing ATP-competitive EGFR TKIs, and are considered a potential therapeutic strategy to overcome treatment resistance in EGFR-mutant patients.

[0006]

[0007] In 2019, Michael J. Eck, Nathanael S. Gray, and Pasi A. from the Dana-Farber Cancer Institute at Harvard Medical School... A team led by JBJ-04-125-02 reported a mutation-selective EGFR allosteric inhibitor. Studies showed it was effective against EGFR L858R / T790M / C797S mutations both in vitro and in vivo. However, JBJ-04-125-02 monotherapy was ineffective in patient-derived cell lines or xenograft models. Subsequently, the team improved JBJ-04-125-02, discovering a new and more effective EGFR allosteric inhibitor, JBJ-09-063. Compared to JBJ-04-125-02, JBJ-09-063 exhibited better pharmacokinetics and efficacy, showing good therapeutic effects against models leading to EGFR-TKI resistance, such as EGFR T790M and C797S mutations. However, further improvements to achieve even better EGFR inhibitory effects remain a challenge. Summary of the Invention

[0008] To address the aforementioned problems, the inventors provide a novel class of oxoisoindole compounds. Compared to JBJ-09-063 and JBJ-04-125-02, these compounds exhibit superior pharmacokinetic properties and EGFR inhibitory effects, demonstrating promising development prospects.

[0009] This invention provides compounds of the following general formula (I) or pharmaceutically acceptable salts thereof:

[0010]

[0011] Ring A can be a substituted or unsubstituted benzene ring, and ring B can be chosen arbitrarily.

[0012] Alternatively, ring A may be a substituted or unsubstituted heterocyclic group or a substituted or unsubstituted C36 cycloalkyl group, and ring B may be chosen from...

[0013]

[0014] Wherein, the substituents on the benzene ring, heterocyclic group, or C36 cycloalkyl group are selected from: C1-4 alkyl groups; n is 1-4;

[0015] R1, R2, R3, R1', R2', R3', and R4' are each independently selected from H, C1-4 alkyl, and deuterated C1-4 alkyl; R4 is H, C2-4 alkyl, or deuterated C2-4 alkyl; when R1, R2, R3, R4, R1', R2', R3', and R4' are substituents, they include monosubstituted, disubstituted, trisubstituted, or tetrasubstituted groups.

[0016] In one embodiment of the present invention, the heterocyclic group comprises:

[0017] In one embodiment of the present invention, the C36 cycloalkyl group comprises:

[0018] In one embodiment of the present invention, when ring A is a benzene ring, ring B may specifically be selected as follows:

[0019] In one embodiment of the present invention, when ring A is a benzene ring, ring B is further preferred:

[0020] In one embodiment of the present invention, when ring A is a substituted or unsubstituted heterocyclic group or a substituted or unsubstituted C36 cycloalkyl group, ring B may specifically be selected as follows:

[0021] In this invention, further preferred compounds of general formula (I) or pharmaceutically acceptable salts thereof are selected from:

[0022]

[0023] The present invention relates to compounds of general formula (I) or pharmaceutically acceptable salts thereof, wherein the pharmaceutically acceptable salt is an inorganic or organic salt, the inorganic salt including hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, sulfonate, benzenesulfonate, and salicylate.

[0024] Another object of the present invention is to provide a pharmaceutical composition comprising the above-described compound of general formula (I) of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

[0025] On the other hand, the present invention provides the use of compounds of the above general formula (I) or pharmaceutically acceptable salts thereof in the preparation of medicaments used as EGFR inhibitors.

[0026] On the other hand, the present invention provides the use of compounds of the above general formula (I) or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating cancer.

[0027] The cancers mentioned in this invention may be selected from: breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer (such as stomach cancer and colorectal cancer), or lung cancer, etc.

[0028] The cancers mentioned in this invention may be selected from: lung cancer, bone cancer, endometrial cancer, skin cancer, head or neck cancer, melanoma of the skin or melanoma of the eye, uterine cancer, ovarian cancer, rectal cancer, hilar cancer, stomach cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney cancer or ureteral cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) cancer, primary central nervous system lymphoma, spinal cord axon cancer, brainstem glioma, pituitary adenoma, etc.

[0029] The beneficial effect of this invention is that it designs a class of compounds with biological functions that inhibit EGFR, thereby providing new means for finding new treatments for cancer, metabolic and immune diseases, cardiovascular diseases and neurological diseases.

[0030] Beneficial effects:

[0031] The present invention provides a class of compounds having the structure shown in general formula (I) for therapeutic and / or prophylactic treatment of patients with cancer, particularly non-small cell lung cancer, who have EGFR mutations T790M / L858RT790M / L858R / C797S, L858R, and / or L858R / C797S, said treatment comprising determining the EGFR activating mutation status of the patient and then administering a compound of formula I as described herein or a pharmaceutical salt thereof to the patient. Detailed Implementation

[0032] The technical solution of the present invention will be described in detail below with reference to the embodiments.

[0033] In this invention, "C1-C6 alkyl" refers to a saturated straight-chain or branched monovalent hydrocarbon group having 1 to 6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, and 2-methyl-2-propyl.

[0034] In this invention, "C3-C6 cycloalkyl" refers to cycloalkyl groups with 3 to 6 carbon atoms respectively.

[0035] In this invention, "heteroaryl" refers to, unless otherwise stated, an unsubstituted or substituted stable 5- or 6-membered monocyclic aromatic ring system, or an unsubstituted or substituted 9- or 10-membered benzo[a]-fused heteroaromatic ring system, or a bicyclic heteroaromatic ring system, which consists of a carbon atom and 1-4 heteroatoms selected from N, O, or S, wherein the nitrogen or sulfur heteroatoms are selectively oxidized and the nitrogen heteroatoms are selectively quaternized.

[0036] In this invention, "substituted" means that one or more hydrogen atoms in a group are replaced by the same or different substituents.

[0037] In this invention, “administering” or “giving” an individual compound means providing the compound of this invention to an individual in need of treatment.

[0038]

[0039] This invention provides a class of novel oxoisoindole compounds or pharmaceutically acceptable salts thereof that can inhibit epidermal growth factor receptor (EGFR) inhibitors, the structural formula of which is shown in general formula (I):

[0040]

[0041] <Pharmaceutical Composition>

[0042] The present invention also provides pharmaceutical compositions comprising the above-described compound of general formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

[0043] The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into solid dosage forms for oral administration, including, but not limited to, capsules, tablets, pills, powders, granules, etc. In these solid dosage forms, the compound of general formula (I) of the present invention is mixed as the active ingredient with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate. Or it may be mixed with the following components: (1) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol and silica; (2) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, gum arabic; (3) humectants, such as glycerin; (4) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginic acid, certain silicates and sodium carbonate; (5) slowing agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate; (8) adsorbents, such as kaolin; (9) lubricants, such as talc, calcium stearate, solid polyethylene glycol, sodium dodecyl sulfate, etc., or mixtures thereof. Buffers may also be included in capsules, tablets and pills.

[0044] The solid dosage forms, such as tablets, sugar pills, capsules, pellets, and granules, can be coated or microencapsulated with coating and shell materials such as enteric coatings and other materials known in the art. They may contain opaque agents, and the release of the active ingredient from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active ingredient may also be formed into microcapsules with one or more of the excipients described above.

[0045] The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, etc. In addition to the compound of general formula (I) or its pharmaceutically acceptable salt as the active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, etc., or mixtures thereof. In addition to these inert diluents, the liquid dosage forms of the present invention may also include conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0046] The suspending agent includes, for example, ethoxylated octadecyl alcohol, polyoxyethylene sorbitol, and dehydrated sorbitol, microcrystalline cellulose, agar, or mixtures thereof.

[0047] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into dosage forms for parenteral injection, including, but not limited to, physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions and dispersions. Suitable carriers, diluents, solvents, and excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0048] The compounds of this invention or pharmaceutically acceptable salts thereof can be formulated into dosage forms for topical administration, including ointments, powders, suppositories, drops, sprays, and inhalers. The compounds of general formula (I) of this invention or pharmaceutically acceptable salts thereof, as active ingredients, are mixed under sterile conditions with a physiologically acceptable carrier and optionally with preservatives, buffers, and propellants, if necessary.

[0049] The pharmaceutical compositions of the present invention comprise a compound of general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier, excipient, and diluent. In preparing the pharmaceutical compositions, the compound of general formula (I) or a pharmaceutically acceptable salt thereof is typically mixed with a pharmaceutically acceptable carrier, excipient, or diluent. The content of the compound of general formula (I) or a pharmaceutically acceptable salt thereof can be 0.01-1000 mg, for example 0.05-800 mg, 0.1-500 mg, 0.01-300 mg, 0.01-200 mg, 0.05-150 mg, 0.05-50 mg, etc.

[0050] <Application>

[0051] The present invention also provides the use of compounds of general formula (I) or pharmaceutically acceptable salts thereof in the preparation for the treatment of cancers in mammals.

[0052] Compounds of formula (I) or pharmaceutically acceptable salts thereof, the compounds and salts of the present invention, are for the treatment of mammals, including humans, for the therapeutic and / or prophylactic treatment of patients with cancer, particularly non-small cell lung cancer, who have EGFR activating mutations, the treatment comprising determining the EGFR activating mutation status of the patient and subsequently administering to the patient a compound of formula I as described herein or a pharmaceutically acceptable salt thereof.

[0053] "Therapeutic effective amount" is the amount of the compound of the present invention that effectively produces a biological or medical response in an individual (e.g., reducing or inhibiting enzyme activity, or improving symptoms, alleviating the condition, slowing or delaying disease progression, or preventing disease).

[0054] The cancers mentioned in this invention include lung cancer, bone cancer, endometrial cancer, skin cancer, head or neck cancer, melanoma of the skin or melanoma of the eye, uterine cancer, ovarian cancer, rectal cancer, hilar cancer, stomach cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney cancer or ureteral cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) cancer, primary central nervous system lymphoma, spinal cord axon cancer, brainstem glioma, pituitary adenoma, etc.

[0055] The compounds of the present invention or pharmaceutically acceptable salts thereof may be administered to mammals, including humans, orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), topically (in powder, ointment, drops) or intratumorally.

[0056] The compounds described in this invention, or pharmaceutically acceptable salts thereof, can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents or other antitumor drugs. This combination therapy can be achieved by administering the individual therapeutic components simultaneously, sequentially, or separately. The therapeutic agents include, but are not limited to: antitumor drugs acting on the chemical structure of DNA, such as cisplatin; antitumor drugs affecting nucleotide synthesis, such as methotrexate and 5-fluorouracil; antitumor drugs affecting nucleic acid transcription, such as doxorubicin, epirubicin, and aclarubicin; antitumor drugs acting on microtubule synthesis, such as paclitaxel and vinorelbine; aromatase inhibitors, such as amylase, letrozole, and renin; and cell signaling pathway inhibitors, such as ALK inhibitors crizotinib, ceritinib, alectinib, and lorlatinib. Antitumor monoclonal antibodies, immunosuppressants such as PD-1 and PD-L1, etc., can also be administered simultaneously or sequentially, in a single formulation or in different formulations. The combination includes not only combinations of one or more other active agents of the compounds of this invention, but also combinations of two or more other active agents of the compounds of this invention.

[0057] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions.

[0058] Example 1:

[0059] 2-(6-(4-cyclohexylphenyl)-1-oxoisoindol-2-yl)-2-(5-fluoro-2-hydroxyphenyl)-N-(thiazolyl-2-yl)acetamide

[0060]

[0061] Preparation of 2-amino-2-(5-fluoro-2-methoxyphenyl)acetonitrile (intermediate 1):

[0062] In a 100 mL thick-walled pressure-resistant bottle, 10 g (65 mmol) of 5-fluoro-2-methoxybenzaldehyde, 0.5 g of anhydrous zinc iodide, and 1 g of 4A molecular sieve were dissolved in 100 mL of anhydrous acetonitrile. The solution was cooled to below -15 °C, and NH3 was bubbled through until saturation. TMSCN (6.4 g, 65 mmol) was added, and the solution was sealed and reacted overnight at 65 °C. After the reaction was complete, the reaction solution was filtered, concentrated, and recrystallized from diethyl ether-petroleum ether to obtain 2-amino-2-(5-fluoro-2-methoxyphenyl)acetonitrile (intermediate 1) (8 g, 68%), a brown oily substance. ESI-MS m / z: 181.2 [M+H] +

[0063] Preparation of 2-amino-2-(5-fluoro-2-methoxyphenyl)acetic acid hydrochloride (intermediate 2):

[0064] 2-Amino-2-(5-fluoro-2-methoxyphenyl)acetonitrile (intermediate 1) (8 g, 44.4 mmol) was dissolved in concentrated hydrochloric acid (100 mL) and reacted overnight at 80 °C. After the reaction was complete, impurities were removed by extraction with ethyl acetate (50 mL). The aqueous phase was concentrated under reduced pressure to give 2-amino-2-(5-fluoro-2-methoxyphenyl)acetic acid hydrochloride (intermediate 2) (6.2 g, 60%), a white solid. ESI-MS m / z: 200.1 [M+H] +

[0065] Preparation of 2-((tert-butoxycarbonyl)amino)-2-(5-fluoro-2-methoxyphenyl)acetic acid (intermediate 3):

[0066] 2-Amino-2-(5-fluoro-2-methoxyphenyl)acetic acid hydrochloride (intermediate 2) (6 g, 25.5 mmol) was dissolved in THF (30 mL) and 1N NaOH (30 mL), then Boc₂O (6.67 g, 30.6 mmol) was added. The mixture was stirred at room temperature for 18 hours. After the reaction was complete, the THF was removed by concentration under reduced pressure. The aqueous phase was washed once with DCM (50 mL), then the pH was adjusted to 2 with 1N HCl, and extracted with DCM (100 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2-((tert-butoxycarbonyl)amino)-2-(5-fluoro-2-methoxyphenyl)acetic acid (intermediate 3) (5.8 g, 76%), a colorless oil. ESI-MS m / z: 300.3 [M+H] +

[0067] Preparation of tert-butyl (1-(5-fluoro-2-methoxyphenyl)-2-oxo-2-(thiazolyl-2-ylamino)ethyl)carbamate (intermediate 4):

[0068] 2-((tert-butoxycarbonyl)amino)-2-(5-fluoro-2-methoxyphenyl)acetic acid (intermediate 3) (5 g, 16.7 mmol) and thiazolium-2-amine hydrochloride (2.7 g, 20.0 mmol) were dissolved in DMF (50 mL), and DIPEA (7 mL, 41.7 mmol) and HATU (9.5 g, 25.0 mmol) were added. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was completed, the reaction mixture was slowly poured into water, extracted with ethyl acetate (100 mL * 3), and the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give tert-butyl(1-(5-fluoro-2-methoxyphenyl)-2-oxo-2-(thiazolium-2-ylamino)ethyl)carbamate (intermediate 4) (4.5 g, 70%), a white solid. ESI-MS m / z: 382.2 [M+H] +

[0069] Preparation of 2-amino-2-(5-fluoro-2-methoxyphenyl)-N-(thiazol-2-yl)acetamide (intermediate 5):

[0070] 4 g (10.5 mmol) of tert-butyl (1-(5-fluoro-2-methoxyphenyl)-2-oxo-2-(thiazol-2-ylamino)ethyl)carbamate (intermediate 4) was dissolved in methanol (30 mL), and 20 mL of HCl (4 M in dioxane) was added. The reaction mixture was stirred at room temperature for 18 hours. After the reaction was completed, the solution was concentrated under reduced pressure to give 2-amino-2-(5-fluoro-2-methoxyphenyl)-N-(thiazol-2-yl)acetamide (intermediate 5) (2.9 g, 98%), a white solid. ESI-MS m / z: 282.1 [M+H] +

[0071] Preparation of 2-(6-bromo-1-oxoisoindol-2-yl)-2-(5-fluoro-2-methoxyphenyl)-N-(thiazolyl-2-yl)acetamide (intermediate 6):

[0072] 2-Amino-2-(5-fluoro-2-methoxyphenyl)-N-(thiazol-2-yl)acetamide (intermediate 5) (2.5 g, 8.9 mmol) and methyl 5-bromo-2-(bromomethyl)benzoate (2.7 g, 8.9 mmol) were dissolved in DMF (40 mL), and DIPEA (6 mL, 35.6 mmol) was added. The mixture was heated to 70 °C and reacted for 2 hours. After the reaction was completed, the mixture was cooled to room temperature, and the reaction mixture was slowly poured into water. The mixture was extracted with ethyl acetate (100 mL * 3), and the organic phases were combined. The mixture was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give 2-(6-bromo-1-oxoisoindol-2-yl)-2-(5-fluoro-2-methoxyphenyl)-N-(thiazol-2-yl)acetamide (intermediate 6) (3 g, 71%), a white solid. ESI-MS m / z: 476.4 [M+H] +

[0073] Preparation of 2-(6-(4-cyclohexylphenyl)-1-oxoisoindol-2-yl)-2-(5-fluoro-2-methoxyphenyl)-N-(thiazolyl-2-yl)acetamide (intermediate 7):

[0074] 2-(6-bromo-1-oxoisoindol-2-yl)-2-(5-fluoro-2-methoxyphenyl)-N-(thiazo-2-yl)acetamide (intermediate 6) (100 mg, 0.21 mmol) and 2-(4-cyclohexylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (72 mg, 0.25 mmol) were dissolved in THF (2 mL), followed by the addition of sodium carbonate (44 mg, 0.42 mmol), PdCl2 (dppf) (15 mg, 0.02 mmol), and water (0.5 mL). The reaction mixture was heated to 70 °C and stirred for 16 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was extracted with ethyl acetate (15 mL * 3), the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give 2-(6-(4-cyclohexylphenyl)-1-oxoisoindol-2-yl)-2-(5-fluoro-2-methoxyphenyl)-N-(thiazol-2-yl)acetamide (intermediate 7) (60 mg, 64%), a white solid. ESI-MS m / z: 556.3 [M + H] +

[0075] Preparation of 2-(6-(4-cyclohexylphenyl)-1-oxoisoindol-2-yl)-2-(5-fluoro-2-hydroxyphenyl)-N-(thiazol-2-yl)acetamide (Example 1):

[0076] 2-(6-(4-cyclohexylphenyl)-1-oxoisoindol-2-yl)-2-(5-fluoro-2-methoxyphenyl)-N-(thiazol-2-yl)acetamide (intermediate 7) (60 mg, 0.11 mmol) was dissolved in DCM (2 mL), and boron tribromide (55 mg, 0.22 mmol) was added. The mixture was stirred at room temperature for 3 hours. After the reaction was completed, the mixture was quenched with water, extracted with DCM (5 mL * 3), and the organic phases were combined. The mixture was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and plate-sected to obtain 2-(6-(4-cyclohexylphenyl)-1-oxoisoindol-2-yl)-2-(5-fluoro-2-hydroxyphenyl)-N-(thiazol-2-yl)acetamide (Example 1) (30 mg, 51%), a white solid. ESI-MS m / z: 542.1 [M+H] +

[0077] 1 H NMR (400MHz, DMSO-d6) δ7.90 (d, J = 0.8Hz, 1H), 7.74-7.68 (m, 1H), 7.62-7.55 (m, 2H), 7.54-7.48(m,2H),7.47-7.40(m,2H),7.12(d,J=6.0Hz,1H),7.01-6.94(m,1H),6.88- 6.78(m,2H),5.85(s,1H),5.82(s,1H),5.11(s,1H),4.92(s,1H),2.58(dq,J=15.0,7. 5Hz,1H),2.25-2.00(m,2H),1.85-1.66(m,3H),1.60-1.47(m,2H),1.46-1.35(m,3H).

[0078]

[0079] Preparation of 4'-bromo-4,4-dimethyl-2,3,4,5-tetrahydro-1,1'-biphenyl (intermediate 8):

[0080] p-Bromophenylboronic acid (1 g, 5 mmol) and 4,4-dimethylcyclohexyl-1-en-1-yltrifluoromethanesulfonate (1.9 g, 7.5 mmol) were dissolved in THF (10 mL) and water (1 mL). Triethylamine (1.4 mL, 10 mmol) and tetrakis(triphenylphosphine)palladium (289 mg, 0.25 mmol) were added. The reaction mixture was heated to 70 °C and stirred for 16 hours under a nitrogen atmosphere. After the reaction was completed, water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 4'-bromo-4,4-dimethyl-2,3,4,5-tetrahydro-1,1'-biphenyl (intermediate 8) (800 mg, 61%), a colorless oil.

[0081] Preparation of 1-bromo-4-(4,4-dimethylcyclohexyl)benzene (intermediate 9):

[0082] 4'-Bromo-4,4-dimethyl-2,3,4,5-tetrahydro-1,1'-biphenyl (intermediate 8) (800 mg, 3.0 mmol) was dissolved in methanol (5 mL), and 10% Pd-C (100 mg) was added. The mixture was stirred at room temperature for 2 hours under H2 atmosphere. After the reaction was completed, the mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain 1-bromo-4-(4,4-dimethylcyclohexyl)benzene (intermediate 9) (0.8 g, 100%), a colorless oil.

[0083] Preparation of 2-(4-(4,4-dimethylcyclohexyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (intermediate 10):

[0084] 1-Bromo-4-(4,4-dimethylcyclohexyl)benzene (intermediate 9) (800 mg, 3.0 mmol) and pinacol diboronate (1100 mg, 4.5 mmol) were dissolved in dioxane (10 mL), and potassium acetate (588 mg, 6.0 mmol) and PdCl2 (dppf) (22 mg, 0.03 mmol) were added. The reaction mixture was heated to 80 °C and stirred for 16 hours under a nitrogen atmosphere. After the reaction was completed, water (30 mL) was added, and the mixture was extracted with ethyl acetate (50 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 2-(4-(4,4-dimethylcyclohexyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (intermediate 10) (400 mg, 42%), a colorless oil.

[0085]

[0086] Preparation of 1-bromo-4-(4-methylcyclohexyl)benzene (intermediate 11):

[0087] Methyltriphenylphosphine bromide (5.9 g, 16.5 mmol) and potassium tert-butoxide (1.8 g, 16.5 mmol) were mixed in 100 mL of diethyl ether and stirred at room temperature for 1 hour. Then, a solution of 4-(4-bromophenyl)cyclohexane-1-one (3 g, 11.8 mmol) in 20 mL of diethyl ether was added dropwise and stirred at room temperature overnight. After the reaction was completed, water (50 mL) was added and the mixture was extracted with 3 x 50 mL of diethyl ether. The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1-bromo-4-(4-methylcyclohexyl)benzene (intermediate 11) (700 mg, 23%), a white solid.

[0088] Preparation of 6-(4-bromophenyl)spiro[2.5]octane (intermediate 12):

[0089] 1-Bromo-4-(4-methylcyclohexyl)benzene (intermediate 11) (700 mg, 2.8 mmol) and pinacol iodomethylborate (1.5 g, 5.6 mmol) were dissolved in toluene (20 mL), and then BEt3 (5.6 mL, 5.6 mmol, 1 M) was added. The mixture was reacted at room temperature for 16 hours. After the reaction was completed, water (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 6-(4-bromophenyl)spiro[2.5]octane (intermediate 12) (300 mg, 40%), a white solid.

[0090] Preparation of 4,4,5,5-tetramethyl-2-(4-(spiro[2.5]octane-6-yl)phenyl)-1,3,2-dioxaborane (intermediate 13):

[0091] The synthesis method is the same as that used for the preparation of intermediate 10.

[0092]

[0093] Preparation of 1-(4-bromophenyl)-4,4-dimethylpiperidine (intermediate 14):

[0094] 1,4-Dibromobenzene (1 g, 4.2 mmol) and 4,4-dimethylpiperidine (475 mg, 4.2 mmol) were dissolved in toluene (20 mL), and BINAP (522 mg, 0.84 mmol), Pd2(dba)3 (384 mg, 0.42 mmol), and sodium tert-butoxide (806 mg, 8.4 mmol) were added sequentially. The mixture was heated to 100 °C for 6 hours under a nitrogen atmosphere.

[0095] After the reaction was complete, water (20 mL) was added, followed by extraction with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give 1-(4-bromophenyl)-4,4-dimethylpiperidine (intermediate 14) (400 mg, 35%), a white solid. ESI-MS m / z: 268.1 [M + H] +

[0096] Preparation of 4,4-dimethyl-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)piperidine (intermediate 15):

[0097] The synthesis method is the same as that used for intermediate 10. ESI-MS m / z: 316 [M+H] +

[0098]

[0099] The synthesis of 6-6-(4-bromophenyl)-6-azaspiro[2.5]octane (intermediate 16) and 6-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)-6-azaspiro[2.5]octane (intermediate 17) is based on the preparation of intermediate 15, wherein 4,4-dimethylpiperidine is replaced by 6-azaspiro[2.5]octane.

[0100]

[0101] Preparation of 4-(4-bromophenyl)-1-cyclopropylpiperidine (intermediate 18):

[0102] 4-(4-bromophenyl)piperidine (2 g, 8.3 mmol) was dissolved in THF (20 mL) and MeOH (20 mL), followed by the addition of (1-ethoxycyclopropoxy)trimethylsilane (2.9 g, 16.6 mmol), sodium cyanoborohydride (782 mg, 12.4 mmol), and acetic acid (744 mg, 12.4 mmol). The reaction mixture was heated to 50 °C and stirred for 16 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 4-(4-bromophenyl)-1-cyclopropylpiperidine (intermediate 18) (1.2 g, 51%), a white solid. ESI-MS m / z: 280.2 [M+H] +

[0103] Preparation of 1-cyclopropyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)piperidine (intermediate 19):

[0104] The synthesis method is the same as that used for intermediate 10. ESI-MS m / z: 328.1 [M+H] +

[0105] Using the corresponding intermediate boron ester instead of 2-(4-cyclohexylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane in Example 1, Examples 2-6 were synthesized following a similar procedure to Example 1 (see Table 1) to obtain the desired products.

[0106] Table 1

[0107]

[0108] Example 7:

[0109] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)phenyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0110]

[0111] Preparation of 4-(4-bromophenyl)-1-methylpiperidin-4-ol (intermediate 20):

[0112] At -78°C, 1,4-dibromobenzene (5 g, 21.2 mmol) was dissolved in THF (100 mL), and n-BuLi (28.5 mL, 1.2 mmol, 2.5 M) was slowly added dropwise. After the addition was complete, the mixture was stirred at -78°C for 1 hour. Then, 1-methylpiperidin-4-one (2.4 g, 21.2 mmol) was added, the mixture was brought to room temperature, and stirred for 1 hour. After the reaction was complete, the mixture was quenched with water, extracted with ethyl acetate (100 mL * 3), and the organic phases were combined. The mixture was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 4-(4-bromophenyl)-1-methylpiperidin-4-ol (intermediate 20) (3.2 g, 56%), a colorless oil. ESI-MS m / z: 270.2 [M + H] +

[0113] Preparation of 4-(4-bromophenyl)-1-methyl-1,2,3,6-tetrahydropyridine (intermediate 21):

[0114] 4-(4-bromophenyl)-1-methylpiperidin-4-ol (intermediate 20) (3 g, 11.1 mmol) was added to 50 mL of hydrochloric acid at 6 N and heated to 100 °C for 16 hours. After the reaction was complete, the mixture was neutralized with Na₂CO₃, extracted with ethyl acetate (100 mL * 3), and the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give 4-(4-bromophenyl)-1-methyl-1,2,3,6-tetrahydropyridine (intermediate 21) (1.6 g, 57%), a white solid. ESI-MS m / z: 252.1 [M + H] +

[0115] Preparation of 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)-1,2,3,6-tetrahydropyridine (intermediate 22):

[0116] The synthesis method is the same as that used for the preparation of intermediate 10. ESI-MS m / z: 300.1 [M+H] +

[0117] Preparation of 2-(5-fluoro-2-methoxyphenyl)-2-(6-(4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)phenyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (intermediate 23):

[0118] The synthesis method is the same as that used for intermediate 7. ESI-MS m / z: 569.3 [M+H] +

[0119] Preparation of 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)phenyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (Example 7):

[0120] The synthesis method is the same as that described in Example 1. White solid. ESI-MS m / z: 555.2 [M+H] +

[0121] 1H NMR (400MHz, DMSO-d6) δ7.93 (d, J = 1.2Hz, 1H), 7.83-7.75 (m, 1H), 7.62-7.56 (m, 1H), 7. 55-7.45(m,5H),7.05(d,J=5Hz,1H),7.03-6.97(m,1H),6.84(s,1H),6.77-6.68(m,1H), 6.31(q,J=6.1Hz,1H),5.89-5.73(m,2H),4.82-4.62(m,1H),4.34-4.11(m,1H),3.65-3. 44(m,2H),2.74(q,J=5.8Hz,1H),2.56-2.51(m,1H),2.46(q,J=5.5Hz,2H),2.36(s,3H).

[0122] Example 8:

[0123] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-(methyl-d3)piperidin-4-yl)phenyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0124]

[0125] Preparation of 4-(4-bromophenyl)-1-(methyl-d3)piperidine (intermediate 24):

[0126] 4-(4-bromophenyl)piperidine (1 g, 4.2 mmol) was dissolved in DCM (20 mL). DIPEA (1.1 mL, 6.3 mmol) and deuterated iodomethane (0.26 mL, 4.2 mmol) were added under ice bath conditions. After the addition was complete, the mixture was reacted under ice bath conditions for 1 hour. After the reaction was complete, water (20 mL) was added, and the mixture was extracted with DCM (20 mL x 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 4-(4-bromophenyl)-1-(methyl-d3)piperidine (intermediate 24) (500 mg, 46%), a white solid. ESI-MS m / z: 257.1 [M+H] +

[0127] Preparation of 1-(methyl-d3)-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)phenyl)piperidine (intermediate 25):

[0128] The synthesis method is the same as that used for intermediate 10. ESI-MS m / z: 305 [M+H] +

[0129] Preparation of 2-(5-fluoro-2-methoxyphenyl)-2-(6-(4-(1-(methyl-d3)piperidin-4-yl)phenyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (intermediate 26):

[0130] The synthesis method is the same as that used for intermediate 7. ESI-MS m / z: 574.3 [M+H] +

[0131] Preparation of 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-(methyl-d3)piperidin-4-yl)phenyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (Example 8):

[0132] The synthesis method is the same as that described in Example 1. White solid. ESI-MS m / z: 560.2 [M+H] +

[0133] 1 H NMR(400MHz,DMSO-d6)δ7.93(d,J=1.2Hz,1H),7.80-7.75(m,1H),7.63-7.55(m,3H),7.53-7.4 8(m,1H),7.47-7.44(m,2H),7.20(d,J=4Hz,1H),7.03(s,1H),6.95-6.90(m,1H),6.90–6.84(m ,1H),5.82(s,1H),5.78(s,1H),4.93(s,1H),4.57(s,1H),2.73(ddd,J=11.5,11.0,5.5Hz,2H) ,2.70-2.60(m,1H),2.41(ddd,J=12.0,11.0,5.5Hz,2H),2.23-2.14(m,2H),2.01-1.87(m,2H).

[0134] Example 9:

[0135] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methylpiperidin-4-yl)cyclohexyl-1-en-1-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0136]

[0137] Preparation of 4-(1,4-dioxopyridine[4.5]dec-7-en-8-yl)pyridine (intermediate 27):

[0138] 4,4,5,5-Tetramethyl-2-(1,4-dioxane[4.5]dec-7-en-8-yl)-1,3,2-dioxaborane (5 g, 18.8 mmol) and 4-bromopyridine (3 g, 18.8 mmol) were dissolved in 1,4-dioxane (50 mL) and water (5 mL). Na2CO3 (4 g, 37.6 mmol) and Pd(PPh3)4 (1.1 g, 0.94 mmol) were added sequentially. The reaction mixture was heated to 100 °C and stirred for 16 hours under a nitrogen atmosphere. After the reaction was complete, the mixture was cooled to room temperature, extracted with water (100 mL) and ethyl acetate (100 mL * 3), the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 4-(1,4-dioxopyridin[4.5]dec-7-en-8-yl)pyridine (intermediate 27) (2.3 g, 57%), a white solid. ESI-MS m / z: 218.1 [M + H] +

[0139] Preparation of 4-(1,4-dioxaacetylpyridine[4.5]dec-8-yl)piperidine (intermediate 28):

[0140] 4-(1,4-dioxopyridin[4.5]dec-7-en-8-yl)pyridine (intermediate 27) (2.2 g, 10.1 mmol) was dissolved in methanol (30 mL) and acetic acid (15 mL), and platinum dioxide (300 mg) was added. The mixture was stirred at room temperature for 48 hours under a hydrogen atmosphere (6 MPa). After the reaction was completed, the mixture was filtered, and the filtrate was neutralized with saturated sodium bicarbonate. The filtrate was extracted with ethyl acetate (100 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 4-(1,4-dioxoacetylpyridin[4.5]dec-8-yl)piperidine (intermediate 28) (1.5 g, 65%), a colorless oil. ESI-MS m / z: 226.2 [M+H] +

[0141] Preparation of tert-butyl 4-(4-oxocyclohexyl)piperidine-1-carboxylate (intermediate 29):

[0142] 4-(1,4-dioxaacetylpyridin[4.5]dec-8-yl)piperidine (intermediate 28) (1.5 g, 6.6 mmol) was dissolved in THF (30 mL), and NaHCO3 (8 mL, 1 M) and Boc2O (1.7 g, 7.9 mmol) were added sequentially. The mixture was stirred at room temperature for 6 hours. After the reaction was completed, water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain tert-butyl 4-(4-oxocyclohexyl)piperidine-1-carboxylate (intermediate 29) (1.5 g, 80%), a colorless oil. ESI-MS m / z: 282.2 [M+H] +

[0143] Preparation of tert-butyl 4-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohexyl-3-en-1-yl)piperidine-1-carboxylate (intermediate 30):

[0144] 1.5 g (5.3 mmol) of tert-butyl 4-(4-oxocyclohexyl)piperidine-1-carboxylate (intermediate 29) was dissolved in 15 mL of THF and slowly added dropwise to a 15 mL solution of NaHMDS (5.3 mL, 5.3 mmol, 1 M) in THF at -78 °C. The mixture was stirred at this temperature for 30 minutes. Then, a 5 mL solution of N-phenylbis(trifluoromethanesulfonyl)imide (1.9 g, 5.3 mmol) in THF was slowly added dropwise. After the addition was complete, the mixture was reacted at -78 °C for 1.5 hours. The mixture was allowed to rise naturally to 0°C. After the reaction was complete, it was quenched with saturated ammonium chloride (50 mL), extracted with ethyl acetate (50 mL * 3), and the organic phases were combined. The mixture was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain tert-butyl 4-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohexyl-3-en-1-yl)piperidin-1-carboxylic acid (intermediate 30) (1.7 g, 77%), a colorless oil. ESI-MS m / z: 414.2 [M + H] +

[0145] Preparation of tert-butyl 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)cyclohex-3-en-1-yl)piperidine-1-carboxylate (intermediate 31):

[0146] The synthesis method is the same as that used for the preparation of intermediate 10. ESI-MS m / z: 392.2 [M+H] +

[0147] Synthesis of 4-(4-(2-(1-(5-fluoro-2-methoxyphenyl)-2-oxo-2-(thiazol-2-ylamino)ethyl)-3-oxoisoindol-5-yl)cyclohex-3-en-1-yl)piperidine-1-carboxylic acid tert-butyl ester (intermediate 32):

[0148] The synthesis method is the same as that used for intermediate 7. ESI-MS m / z: 661.1 [M+H] +

[0149] Preparation of 2-(5-fluoro-2-hydroxyphenyl)-2-(1-oxo-6-(4-(piperidin-4-yl)cyclohex-1-en-1-yl)isoindol-2-yl)-N-(thiazo-2-yl)acetamide (intermediate 33):

[0150] The synthesis method is the same as that described in Example 1. ESI-MS m / z: 547.1 [M+H] +

[0151] Preparation of 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methylpiperidin-4-yl)cyclohex-1-en-1-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (Example 9):

[0152] 4-(4-(2-(1-(5-fluoro-2-methoxyphenyl)-2-oxo-2-(thiazol-2-ylamino)ethyl)-3-oxoisoindol-5-yl)cyclohex-3-en-1-yl)piperidin-1-carboxylic acid tert-butyl ester (intermediate 33) (200 mg, 0.36 mmol) was dissolved in methanol (5 mL), and paraformaldehyde (49 mg, 0.54 mmol) and acetic acid (22 mg, 0.36 mmol) were added sequentially. The reaction mixture was stirred at room temperature for 2 hours, and then sodium cyanoborohydride (23 mg, 0.36 mmol) was added. The mixture was stirred at room temperature for 16 hours. After the reaction was completed, the solution was concentrated under reduced pressure and separated by preparative chromatography to obtain 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methylpiperidin-4-yl)cyclohexyl-1-en-1-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (Example 9) (30 mg, 15%), a white solid. ESI-MS m / z: 561.2 [M+H] +

[0153] 1H NMR (400MHz, DMSO-d6) δ7.65 (s, 1H), 7.30 (d, J = 4.0Hz, 1H), 7.27-7.24 (m, 1H), 7. 20-7.16(m,1H),7.11(d,J=4.0Hz,1H),7.02-6.97(m,1H),6.82(s,1H),6.80-6.77 (m,1H),6.15(q,J=6.2Hz,1H),5.83-5.80(m,1H),5.78-5.71(m,1H),4.63-4.56( m,1H),3.96-3.87(m,1H),2.77-2.68(m,2H),2.56-2.36(m,5H),2.25(s,3H),2.18 -2.09(m,1H),2.02-1.85(m,3H),1.50-1.41(m,1H),1.39-1.26(m,3H),1.26-1.17(m,1H).

[0154] Example 10:

[0155] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methylpiperidin-4-yl)cyclohexyl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0156]

[0157] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methylpiperidin-4-yl)cyclohexyl-1-en-1-yl)-1-oxoisoindol-2-yl)-N-(thiazol-2-yl)acetamide (Example 9) (100 mg, 0.18 mmol) was dissolved in methanol (5 mL), and Pd-C (50 mg) was added. The mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. After the reaction was completed, the mixture was filtered through diatomaceous earth, and the filtrate was concentrated under pressure. 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(4-(1-methylpiperidin-4-yl)cyclohexyl)-1-oxoisoindol-2-yl)-N-(thiazol-2-yl)acetamide (Example 10) (55 mg, 55%) was obtained as a white solid. ESI-MS m / z: 563.2 [M+H] +

[0158] 1H NMR(400MHz, DMSO-d6)δ7.86(s,1H),7.59(d,J=4.0Hz,1H),7.40-7.34(m,1H),7.34(s,1H),7.12(d,J=4.0Hz,1H),7.01-6.92( m,1H),6.89-6.76(m,2H),5.85(s,2H),5.08(s,1H),4.80-4.45(m,1H),2.92-2.58(m,3H),2.45-2.33(m,2H),2.25(s,3H),2.16 -2.08(m,2H),2.00-1.93(m,2H),1.87-1.76(m,2H),1.66-1.55(m,2H),1.39-1.20(m,4H),1.10-0.96(m,2H).

[0159]

[0160] Preparation of tert-butyl 4-(4-bromo-2-oxopyridin-1(2H)-yl)piperidine-1-carboxylate (intermediate 34):

[0161] 4-((methanesulfonyl)oxy)piperidin-1-carboxylic acid tert-butyl ester (2 g, 7.2 mmol) was dissolved in DMF (20 mL), followed by the addition of 4-bromopyridin-2(1H)-one (1.2 g, 7.2 mmol) and cesium carbonate (3.5 g, 10.8 mmol). The mixture was then heated to 100 °C for 3 hours. After the reaction was complete, the mixture was cooled to room temperature, extracted with water (100 mL) and ethyl acetate (100 mL x 3), and the organic phases were combined. The mixture was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 4-(4-bromo-2-oxopyridin-1(2H)-yl)piperidin-1-carboxylic acid tert-butyl ester (intermediate 34) (1.3 g, 52%), a white solid. ESI-MS m / z: 357.1 [M+H] +

[0162] Preparation of tert-butyl 4-(2-oxo-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)pyridin-1(2H)-yl)piperidine-1-carboxylate (intermediate 35):

[0163] The synthesis method is the same as that used for intermediate 10. ESI-MS m / z: 405.2 [M+H] +

[0164] Example 11:

[0165] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(1-(1-methylpiperidin-4-yl)-2-oxo-1,2-dihydropyridin-4-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0166]

[0167] Example 11 was synthesized according to the preparation method of Example 9, wherein intermediate 35 replaced intermediate 31; it was a white solid. ESI-MS m / z: 574.2 [M+H] +

[0168] 1 H NMR (400MHz, DMSO-d6) δ7.81 (s, 1H), 7.68 (d, J = 4.0Hz, 1H), 7.65-7.61 (m, 1H), 7 .60-7.58(m,1H),7.57(s,1H),7.15-7.10(m,1H),7.00-6.94(m,1H),6.89-6.82 (m,2H),6.01-5.85(m,2H),5.82(s,2H),5.02-4.90(m,2H),3.87(tt,J=14.5,5. 7Hz,1H),2.90-2.68(m,4H),2.42-2.33(m,2H),2.31(s,3H),1.68-1.20(m,2H).

[0169] Example 12:

[0170] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(1-(1-methylpiperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0171]

[0172] Preparation of 8-(1-methylpiperidin-4-yl)-1,4-dioxa-8-azaspiro[4.5]decane (intermediate 36):

[0173] 1-Methylpiperidin-4-one (2 g, 17.7 mmol) and 1,4-dioxa-8-azaspiro[4.5]decane (2.5 g, 17.7 mmol) were dissolved in 1,2-dichloroethane (50 mL), and acetic acid (1.1 g, 17.7 mmol) and sodium triacetoxyborohydride (5.6 g, 26.5 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was directly concentrated under reduced pressure and purified by column chromatography to obtain 8-(1-methylpiperidin-4-yl)-1,4-dioxa-8-azaspiro[4.5]decane (intermediate 36) (2.5 g, 59%), a pale yellow liquid. ESI-MS m / z: 241.1 [M+H] +

[0174] Preparation of 1'-methyl-[1,4'-piperidin-4-one (intermediate 37):

[0175] 8-(1-methylpiperidin-4-yl)-1,4-dioxa-8-azaspiro[4.5]decane (intermediate 36) (2.5 g, 10.4 mmol) was added to concentrated hydrochloric acid (30 mL) and stirred at room temperature for 2 hours. After the reaction was complete, the pH was adjusted to 14 with sodium hydroxide, and the mixture was extracted with ethyl acetate (100 mL * 3). The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 1'-methyl-[1,4'-piperidin-4-one (intermediate 37) (1.4 g, 70%), a colorless liquid. ESI-MS m / z: 197.2 [M+H] +

[0176] Synthesis of 1-(1-methylpiperidin-4-yl)-1,2,3,6-tetrahydropyridine-4-yl trifluoromethanesulfonate (intermediate 38):

[0177] The synthesis method is the same as that used for intermediate 30. ESI-MS m / z: 329.3 [M+H] +

[0178] Preparation of 1-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1,2,3,6-tetrahydropyridine (intermediate 39):

[0179] The synthesis method is the same as that used for intermediate 10. ESI-MS m / z: 307.2 [M+H] +

[0180] Preparation of 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(1-(1-methylpiperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (intermediate 40):

[0181] The synthesis method is the same as that used for intermediate 7. ESI-MS m / z: 562.2 [M+H] +

[0182] Preparation of 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(1-(1-methylpiperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide (Example 12):

[0183] The synthesis method is the same as that described in Example 1. White solid, ESI-MS m / z: 562.2 [M+H] +

[0184] 1 H NMR(400MHz,DMSO-d6)δ7.62(s,1H),7.38-7.29(m,1H),7.23-7.19(m,1H),7.19-7.11(m,2H),7.04-6.93(m, 1H),6.84(s,1H),6.81-6.77(m,1H),6.32(q,J=6.2Hz,1H),5.97-5.62(m,2H),4.67-4.43(m,1H),4.14-3.92( m,1H),3.70-3.39(m,2H),2.90-2.80(m,1H),2.72(ddd,J=11.5,11.0,5.5Hz,2H),2.58-2.49(m,3H),2.39(dd d,J=12.0,11.0,5.5Hz,2H),2.25(s,3H),2.13(dt,J=15.2,7.7Hz,1H),1.99-1.87(m,2H),1.80-1.63(m,2H).

[0185] Example 13:

[0186] 2-(5-fluoro-2-hydroxyphenyl)-2-(6-(1'-methyl-[1,4'-piperidinyl]-4-yl)-1-oxoisoindol-2-yl)-N-(thiazolyl-2-yl)acetamide

[0187]

[0188] The synthesis method is the same as that used in Example 10. White solid, ESI-MS m / z: 564.2 [M+H] +

[0189] 1H NMR(400MHz,DMSO-d6)δ7.81(s,1H),7.61-7.54(m,1H),7.39(s,1H),7.35-7.31(m,1H),7.13 (s,1H),6.97(d,J=4.2Hz,1H),6.86-6.76(m,2H),5.83(s,2H),5.13-4.97(m,1H),4.84-4.59( m,1H),2.87-2.77(m,3H),2.71(ddd,J=11.5,11.0,5.5Hz,2H),2.47-2.32(m,4H),2.26(s,3H) ,2.19-2.06(m,3H),2.00-1.89(m,2H),1.88-1.78(m,2H),1.73(ddt,J=16.6,7.8,5.6Hz,2H).

[0190] Example 14: Biological assay analysis

[0191] I. Evaluation of the inhibitory effect of the compounds of this invention on EGFR kinase:

[0192] Measurement process:

[0193] The IC50 of the obtained compound kinase was determined using the migration rate alteration method, with JBJ-04-125-02 and JBJ-09-063 as positive controls. Experimental reagents are shown in Table 2, and experimental instruments are shown in Table 3.

[0194] Table 2 Experimental Reagents

[0195] Reagent Name Item number Supplier EGFR L858R / T790M / C797S 08-115 Carna Caliper substrate 18 114202 GL 384-well plate 3573 Corning Dimethyl sulfoxide D8418-1L Sigma

[0196] Table 3 Experimental Instruments

[0197]

[0198]

[0199] Experimental steps:

[0200] 1) Preparation of compound solution: The weighed compound was dissolved in dimethyl sulfoxide (DMSO) in an EP tube to prepare a 10 mM stock solution, which was then stored at -20°C in the dark for later use. After diluting the stock solution to the required concentration for the experiment, 250 nL was transferred to a 384-well plate using an Echo 550.

[0201] 2) Preparation of kinase buffer and kinase solution: Prepare kinase buffer and use kinase buffer to prepare kinase solution. Add kinase solution to the experimental wells, positive control wells, and blank control wells, and add kinase buffer to the negative control wells. After adding, centrifuge and incubate at room temperature for 10 minutes.

[0202] 3) Initial reaction: Prepare a mixed solution of ATP and kinase substrate using kinase buffer, add the mixed solution to start the reaction, centrifuge after addition, and incubate at room temperature for 40 minutes.

[0203] 4) Termination of reaction: Add stop solution to terminate the reaction and measure the conversion rate.

[0204] 5) Inhibition rate and IC50 calculation: Process the data using GraphPad Prism 5 and calculate the inhibition rate and IC50 values.

[0205]

[0206] Table 4. Kinase inhibitory activity of the tested compounds

[0207] test substance <![CDATA[Test substance IC 50 (nmol / L)]]> JBJ-04-125-02 1.2 JBJ-09-063 0.87 Example 1 0.65 Example 2 0.71 Example 3 0.35 Example 4 0.72 Example 5 0.67 Example 6 0.59 Example 7 0.81 Example 8 0.85 Example 9 0.54 Example 10 0.63 Example 11 0.58 Example 12 0.51 Example 13 0.44

[0208] II. Cancer cell model determination

[0209] HTRF Phospho EGFR assay: Cell line and culture medium: BaF3 cell line (EGFR L858R / T790M / C797S mutant, a common mutation type in non-small cell lung cancer). Cells were maintained at 37°C and 5% CO2 in RPMI ATCC supplemented with 10% fetal bovine serum (FBS) (Gibco) + 2mM glutamine + 0.5μg / ml puromycin.

[0210] Operating procedures:

[0211] As described above, after prefilling a Greiner BioOne, Nr. 78408 microtiter plate with 12.5 nmol of a DMSO solution containing the test compound (dose-response) or DMSO alone, cells were transferred to the microtiter plate at a rate of 20,000 cells / well and 12.5 μl of growth medium / well. After rotating the plate at 300 x g for 30 seconds, the cells were incubated at 37°C, 5% CO2, and 95% humidity for 4 hours. Cell lysis was performed by adding 4 μl / well of supplemental lysis buffer (Cisbio, Phospho EGFRHTRF kit, 64EG1PEH) to the compound mixture, followed by incubation at room temperature with shaking (400 rpm) for 30 minutes. The plate was then frozen and stored overnight at 80°C. The following day, after thawing the plate, 4 μl of a mixture of anti-Phospho EGFR compound and anti-Phospho EGFR d2 antibody prepared in the supplied assay buffer was added to each well. The covered plate was then incubated at room temperature for 4 hours, after which the fluorescence emission at 616 and 665 nm was read using an Envision plate reader (Perkin Elmer). The data were analyzed in a similar manner to the above, using a normalization ratio of 10000 multiplied by the 665 to 616 signals.

[0212] The results are shown in Table 5.

[0213] Table 5. HTRF, Phospho EGFR, and TMLRCS assay data for H1975 and BaF3 cells.

[0214] test substance <![CDATA[BaF3IC 50 (nmol / L)]]> JBJ-04-125-02 50 JBJ-09-063 15 Example 1 10 Example 2 9.5 Example 3 5.3 Example 4 11 Example 5 8.2 Example 6 12 Example 7 10.5 Example 8 12 Example 9 7 Example 10 10 Example 11 9.6 Example 12 7.2 Example 13 6.5

[0215] The results showed that all compounds in this invention are highly effective EGFR inhibitors.

[0216] For compounds of general formula (I), the linking and substituent groups have a significant impact on the pharmacodynamic properties of the compound. Although the invention has been described through specific embodiments above, it should not be construed as limiting; rather, the invention covers the general aspects previously disclosed. Various modifications and embodiments are possible without departing from the spirit and scope of the invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof with the structure shown in general formula (I): The specific structure of the compound is selected from: 。 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The pharmaceutically acceptable salt is an inorganic or organic salt; the inorganic salt includes hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, and salicylate.

3. A pharmaceutical composition, characterized in that, It contains the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

4. Use of the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament used as an EGFR inhibitor.

5. Use of the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer.

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