MET protein degradation-inducing compounds

Abstract

The present invention relates to compounds that degrade target proteins based on chaperone-mediated protein degradation (CMPD) technology, their production methods, and their uses. The present invention provides target protein degradation compounds that include a first moiety (chaperone binding, CB) capable of binding to a chaperone protein or protein component of a chaperone complex and a second moiety (target binding, TB) capable of binding to a target protein or protein. Such compounds are compounds or stereoisomers thereof that are configured in the form of a chaperone binding moiety, a linker (L), and a target protein binding moiety. The compounds of the present invention have the effect of inducing MET protein degradation based on CMPD technology. Therefore, the compounds of the present invention are expected to have excellent anticancer effects that can overcome resistance and mutation of targeted anticancer drugs by completely degrading the target protein (MET).

Description

[Technical Field]

[0001] The present invention relates to compounds that degrade target proteins based on chaperone-mediated protein degradation (CMPD) technology, as well as methods for producing the same and uses thereof. [Background technology]

[0002] Cancer remains one of the most elusive diseases since the first therapeutic agents were developed in the 1940s. As cancer treatment technology advances, the form of anticancer drugs is also changing. Approximately 40% of new drug R&D investments are focused on anticancer drugs, and many new anticancer drugs have been released. However, unmet demand among cancer patients remains. The current direction of anticancer drug development is to develop new anticancer drugs that overcome resistance or improve therapeutic response rates. Existing anticancer drugs rely on existing technologies, such as small molecule compounds and antibodies. Only 400 of the approximately 3,000 disease-causing genes have been approved for treatment (85% of targets are undruggable). Even drugs developed for cancer treatment have issues with side effects, resistance, and response rates, making the introduction of next-generation drugs and technologies crucial.

[0003] Due to the limitations of existing drug development technologies that focus on inhibiting the function of disease-related proteins, "targeted protein degradation technology (PDT)" has emerged as a new strategy to overcome these challenges. Targeted protein degradation is a novel concept that selectively degrades proteins and removes disease-related proteins themselves. Chemically induced targeted protein degradation is a novel method for the drug development of small molecule compounds. Small molecule compounds promote the interaction of various cellular protein degradation pathway components with target proteins or proteins, and are used as a disease treatment method to induce the degradation of the target proteins or proteins. This technology can be applied to diseases that target proteins that are undruggable by existing drug development technologies. It is expected that PDT will not only degrade and essentially remove disease-causing proteins, but also overcome the side effects and resistance of previously developed drugs.

[0004] In the field of PDT, proteolysis-targeting chimeras (PROTACs), a method using E3 ligases, are a platform that utilizes small molecules to induce specific protein degradation. PROTAC molecules are bifunctional small molecules that simultaneously bind to target proteins and E3-ubiquitin ligases. Therefore, PROTAC molecules trigger polyubiquitination and subsequent degradation of the target protein by the proteasome. PROTAC molecules have good tissue distribution and the ability to target intracellular proteins. However, PROTAC molecules face the problem of resistance to PROTACs through E3 ligase mutations. Therefore, PROTAC technology requires the discovery of novel E3 ligases that are suitable for targeting new proteins with specific substrate specificity.

[0005] On the other hand, the CMPD technology of the present invention overcomes the limitations of existing PROTAC technology by promoting and inducing protein degradation in vivo through the ubiquitin proteasome system (UPS pathway) via interactions with various E3 ligases, rather than through the direct involvement of E3 ligases, via a chaperone complex.

[0006] In addition, when a disease-related target protein is a client protein of a chaperone complex, it can be selectively targeted for degradation, which allows for a wider range of target proteins and greater scalability compared to PROTACs. This makes it possible to study a variety of diseases, including cancer, degenerative brain diseases, and rare diseases, and also facilitates pipeline scalability.

[0007] Lung cancer is a common cancer worldwide and a leading cause of cancer-related deaths. While the five-year survival rate has improved since the 2000s, it still remains at 28.2%. In cases involving distant metastasis, the observed survival rate in Korea is a very low 6.1%. Non-small cell lung cancer (NSCLC) is a cancer characterized by a variety of genetic mutations. Despite the development of numerous treatments, there remains significant unmet need. This has led to various research efforts, including the development of treatments targeting resistance mutations and combination therapy. Approximately 30% of all NSCLC patients are diagnosed with epidermal growth factor receptor (EGFR) mutations. The market for treatments targeting this disease has focused on first-generation EGFR inhibitors and second-generation treatments for the EGFR T790M mutation, which develops as a result of resistance to first-generation treatments. Among all non-small cell lung cancer patients, those with c-MET (hepatocyte growth factor receptor) mutations account for 6% and are an uncommon type. However, there are approximately 250,000 patients with c-MET mutations, which are the main cause of resistance after first- and second-generation EGFR therapies. As a result, expectations and demand for MET-targeted therapeutic agents are increasing in the non-small cell lung cancer treatment market.

[0008] The present invention provides a new alternative to targeted protein degradation in MET-mutated non-small cell lung cancer disease through the development of CMPD-based drugs targeting the MET protein.

[0009] The present inventors designed the structure of CMPD drugs by using various linkers to CMPD-derived compounds, which are binding moieties of chaperone complex-associated proteins, and existing target drugs (e.g., crizotinib, capmatinib, tepotinib, etc.) as target-binding substances. As a result, we found that MET, a major target protein of interest in non-small cell lung cancer, can be degraded and removed at its source by CMPD, and that this can be expected to have superior anticancer effects compared to existing anticancer drugs due to the complete degradation of the target protein MET. This led to the completion of the present invention. Summary of the Invention [Problem to be solved by the invention]

[0010] An object of the present invention is to provide compounds that degrade proteins via CMPD.

[0011] Another object of the present invention is to provide a method for producing a compound that degrades proteins by CMPD.

[0012] Another object of the present invention is to provide uses of compounds that degrade proteins by CMPD. [Means for solving the problem]

[0013] The present invention provides compounds that degrade target proteins based on CMPD technology. Specifically, the present invention provides target protein degradation compounds that include a first moiety (Chaperone binding, CB) capable of binding to a chaperone protein or protein component of a chaperone complex, and a second moiety (Target binding, TB) capable of binding to a target protein or protein.

[0014] The compound according to the present invention is a compound represented by the following chemical formula I or a stereoisomer thereof, which is composed of a chaperone binding moiety (hereinafter referred to as "CB")-linker (L)-target protein binding moiety (hereinafter referred to as "TB"):

[0015] <Chemical formula I> CB-L-TB CB is a first moiety capable of binding to a chaperone protein or protein component of a chaperone complex; JPEG2025539164000002.jpg39150, which exists in meta or para form.

[0016] The chaperone complex of the present invention may be composed of other cofactors or cochaperones selected from HSP90 (heat shock protein 90), HSP70 (heat shock protein 70), IAP (inhibitor of apoptosis protein), E3 ligase (e.g., CHIP (Carboxyl Terminus of HSC70-interacting Protein)), HECTD3 (Homologous to E6AP C-Terminus domain containing 3), and CUL5 (cullin 5).

[0017] TB is a target protein or a second moiety capable of binding to a protein; JPEG2025539164000003.jpg50131, and preferably JPEG2025539164000004.jpg41154.

[0018] The target protein according to the present invention may be selected from target drugs such as crizotinib, savolitinib, cabozantinib, capmatinib, and tepotinib.

[0019] L is a linker connecting a first moiety (CB) capable of binding to a chaperone protein or protein component of a chaperone complex and a second moiety (TB) capable of binding to a target protein or protein; JPEG2025539164000005.jpg31133, and preferably JPEG2025539164000006.jpg37157.

[0020] In the present invention, the linker is non-degradable in vivo and does not interfere with the ability of the chaperone complex-associated protein binding substance (Chaperone binding) and the target protein binding substance (Target binding) to bind to each other.

[0021] The present invention can produce compounds of Formula I having the structures set forth in Table 1 below.

[0022] [Table 1] TIFF2025539164000008.tif94123

[0023] The present invention also provides a method for preparing the compound represented by formula I.

[0024] In the present invention, the compound represented by chemical formula I is prepared by the following steps: i) preparing a first moiety (CB), which is an acid (aicd) intermediate in the meta or para form of CB, capable of binding to a chaperone protein or protein component of a chaperone complex; ii) preparing an L-TB intermediate by binding a second moiety (TB), capable of binding to a target protein or protein, to a linker; and iii) coupling the CB intermediate prepared in step i) with the L-TB intermediate prepared in step ii) to prepare the compound CB-L-TB represented by chemical formula I. In step i), compound (a) of the following Chemical Formula 1 is reacted with compound (b) of Chemical Formula 2 to prepare compound (c) of Chemical Formula 3, which is then hydrated to prepare intermediate compound (d) of Chemical Formula 4, or compound (g) of the following Chemical Formula 5 is reacted with compound (b) of Chemical Formula 2 to prepare compound (h) of Chemical Formula 6, which is then hydrated to prepare intermediate compound (i) of Chemical Formula 7.

[0025] <Chemical formula 1> JPEG2025539164000009.jpg35106<Chemical formula 2> JPEG2025539164000010.jpg36117<Chemical formula 3> JPEG2025539164000011.jpg37118<Chemical formula 4> JPEG2025539164000012.jpg40122<Chemical formula 5> JPEG2025539164000013.jpg37118<Chemical formula 6> JPEG2025539164000014.jpg36114<Chemical formula 7> JPEG2025539164000015.jpg33123 The step ii) is a target drug JPEG2025539164000016.jpg45133 and the linker JPEG2025539164000017.jpg33135 to produce L-TB, an intermediate in which the target drug and linker are bound.

[0026] Step iii) comprises coupling the intermediate compound (d) of Chemical Formula 4 or the intermediate compound (i) of Chemical Formula 7 prepared in step i) with the intermediate L-TB prepared in step ii) to prepare the compound CB-L-TB represented by Chemical Formula I.

[0027] The present invention also provides uses of the compounds represented by formula I.

[0028] The compounds of formula I according to the present invention are generally useful as anti-cancer therapies, and therefore the disease state treated by the compounds of the present invention is cancer. The terms "cancer" or "tumor" are well known in the art and refer to the presence of cells in a subject that have typical characteristics of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, reduced cell death / apoptosis, and certain characteristic morphological features. Cancer cells are usually in the form of solid tumors. However, cancer also includes non-solid tumors, such as blood tumors, such as leukemia, where cancer cells originate from the bone marrow. As used herein, the term "cancer" includes not only pre-malignant but also malignant cancers.Cancers include acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia (monocytic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic), acute T-cell leukemia, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchioloalveolar carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, chronic myeloid leukemia, colorectal cancer, colon cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt's lymphoma lymphoma), dysproliferative changes (dysplasias and metaplasias), embryonal carcinoma, endometrial carcinoma, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal carcinoma, estrogen receptor positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, heavy chain disease, hemangioblastoma, liver tumor, hepatocellular carcinoma, hormone refractory prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin's lymphoma and non-Hodgkin's lymphoma), malignant tumors and hyperproliferative disorders of the bladder, breast, colon, lung, ovary, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, These include, but are not limited to, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung carcinoma, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovium, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular cancer, uterine cancer, and Wilms' tumor.Other cancers include primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelial cancer, gynecological cancer, uterine cancer, gestational trophoblastic disease, male reproductive cancer, seminal vesicle cancer, testicular cancer, germ cell tumors, endocrine tumors, thyroid cancer, adrenal cancer, pituitary cancer, hemangioma, sarcoma derived from bone and soft tissue, Kaposi's sarcoma sarcoma), nerve cancer, eye cancer, meningeal cancer, glioblastoma, neuroma, neuroblastoma, Schwannomas, hematopoietic malignancies, such as solid tumors resulting from leukemia, metastatic melanoma, recurrent or persistent epithelial ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumors, colon cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small cell lung cancer, malignant glioma, epithelial ovarian cancer, primary peritoneal serous carcinoma, metastatic liver cancer, neuroendocrine cancer, refractory malignancies, triple-negative breast cancer, HER2-amplified breast cancer, Includes nasopharyngeal carcinoma, oral cancer, cholangiocarcinoma, hepatocellular carcinoma, squamous cell carcinoma of the head and neck (SCCHN), non-medullary thyroid carcinoma, recurrent glioblastoma multiforme, neurofibromatosis type 1, central nervous system cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumors, triple-negative breast cancer, colon cancer, sarcoma, melanoma, kidney cancer, endometrial cancer, thyroid cancer, rhabdomyosarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumor, mantle cell lymphoma, and refractory malignancies.

[0029] The pharmaceutical compositions of the present invention can be formulated by conventional methods into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injectable solutions. When formulated, they are prepared using commonly used diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, and the like. These solid dosage forms can be prepared by mixing the compound with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid dosage forms include suspensions, liquids, emulsions, syrups, and the like. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, can also be used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases that can be used include witepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin. Any administration route is conceivable, including oral, rectal, intravenous, intramuscular, subcutaneous, intrabronchial inhalation, intrauterine dural, or intracerebrovascular injection.

[0030] In addition, the dosage of the pharmaceutical composition according to the present invention may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc. The specific dosage and treatment regimen for a particular patient will vary depending on various factors, including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, excretion rate, drug combination, the judgment of the treating physician, and the severity of the particular disease being treated. Therefore, the dosages described above are not intended to limit the scope of the present invention in any way. [Effects of the Invention]

[0031] The compounds of the present invention have the effect of inducing degradation of MET protein based on CMPD technology, and therefore, are expected to have excellent anticancer effects that can overcome resistance and mutation of targeted anticancer drugs by completely degrading the target protein (MET). [Brief explanation of the drawings]

[0032] [Figure 1] 1 shows the time-dependent degradation of target proteins in H596 cells by the compound OZD-MET 01 of the present invention.

[0033] [Figure 2] 1 shows the tendency of target protein degradation in H596 cells by the compound of the present invention, OZD-MET 01, at varying doses.

[0034] [Figure 3] 1 shows the time-dependent degradation of target proteins in H1437 cells by the compound OZD-MET 01 of the present invention.

[0035] [Figure 4] 1 shows the tendency of target protein degradation in H1437 cells by the compound of the present invention, OZD-MET 01, at varying doses.

[0036] [Figure 5] 1 shows the tendency of target protein degradation in H596 cells after 72 hours at a dose of 5 μM by compounds OZD-MET 01 to 11 of the present invention.

[0037] [Figure 6] 1 shows the tendency of target protein degradation in H1437 cells after 72 hours at a dose of 5 μM by compounds OZD-MET 01 to 11 of the present invention.

[0038] [Figure 7]FIG. 1 shows the reduction and restoration of target protein expression in H596 cells by the compound of the present invention, OZD-MET 01.

[0039] [Figure 8] 1 shows the reduction and restoration of target protein expression in H1437 cells by the compound of the present invention, OZD-MET 01.

[0040] [Figure 9] FIG. 1 shows the reduction and restoration of target protein expression in H596 cells by the compound of the present invention, OZD-MET 02.

[0041] [Figure 10] FIG. 1 shows the reduction and restoration of target protein expression in H1437 cells by the compound of the present invention, OZD-MET 02.

[0042] [Figure 11] 1 shows the reduction and restoration of target protein expression in H596 cells by the compound of the present invention, OZD-MET03.

[0043] [Figure 12] 1 shows the reduction and restoration of target protein expression in H1437 cells by the compound of the present invention, OZD-MET03. DETAILED DESCRIPTION OF THE INVENTION

[0044] The present invention will be described in more detail below using examples and experimental examples. However, these examples and experimental examples are intended to exemplify the present invention and are not intended to limit the scope of the present invention.

[0045] Example 1 Preparation of 2-{2-[(3-{[1-(2,4-dihydroxy-5-isopropylphenyl)-N-methylformamido]methyl}phenyl)formamido]ethoxy}ethyl 4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}pyrazol-1-yl)piperidine-1-carboxylate (hereinafter referred to as "OZD-MET 01") JPEG2025539164000018.jpg311641) Preparation of intermediate compound (d) (meth) Methyl 3-formylbenzoate is dissolved in methanol, methylamine is added, and sodium borohydride is added and stirred at 0° C. After all the starting materials have reacted, the methanol is removed under reduced pressure, and the mixture is dissolved in methylene chloride, washed with a small amount of brine (saturated aqueous sodium chloride solution), dried over MgSO4, filtered, and the solvent is removed under reduced pressure to obtain compound (a) below, which is used immediately in the next reaction.

[0046] 2,4-Dihydroxy-5-isopropylbenzoate was dissolved in methanol: THF (tetrahydrofuran): HO (1:1:1), and then lithium hydroxide was added and the mixture was allowed to react for 12 hours. After the entire reaction, the mixture was filtered through Celite 545, and the solvent was removed from the filtrate under reduced pressure. After dissolving with HO, the mixture was acidified with hydrochloric acid, extracted twice with EA (ethylacetate), washed with brine, dried over MgSO, filtered, and the solvent was removed under reduced pressure to obtain the following compound (b) in 97% yield.

[0047] Compounds (a) and (b) were dissolved in dimethylformamide, EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide), HOBT (hydroxybenzotriazole), and DIPEA (N,N-diisopropylethylamine) were added, and the mixture was stirred at room temperature for 12 hours to complete the reaction. NaHCO3 (aq.) (aqueous sodium bicarbonate) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure. Compound (c) was then separated using a silica gel column, affording a clear liquid in 86% yield.

[0048] Compound (c) was dissolved in methanol:THF:HO (1:1:1), and then lithium hydroxide was added and the mixture was allowed to react at room temperature for 15 hours. The solvent was removed, and the residue was dissolved in HO. The mixture was then acidified with hydrochloric acid, extracted with EA, washed with brine, and dried with MgSO to obtain solid compound (d) in a 99% yield.

[0049] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 2.81(s, 3H), 3.19(m, 1H), 4.39(t, 2H), 6.41(s, 1H), 7.40-7.61(m, 3H), 8.19(m, 1H), 8.46(m, 1H), 10.07(s, 1H) JPEG2025539164000019.jpg43162

[0050] 2) Preparation of Crizotinib-Linker Intermediate Compound (f) Carbonyldiimidazole (CDI) was dissolved in methylene chloride and cooled to 0°C. Then, (R)-crizotinib and 2-[2-(hydroxyethoxy)ethyl]-1H-isoindole-1,3(2H)-dione were added and reacted for 10 hours. After all the starting materials had reacted, the methylene chloride was removed to obtain the following compound (e).

[0051] Compound (e) was dissolved in ethanol, and then hydrazine hydrate was added thereto. The mixture was refluxed at 110°C for 1 hour, and then stirred at room temperature. The resulting white solid was filtered, yielding the following compound (f) in a 66% yield.

[0052] 1 H NMR (500MHz, DMSO) δ1.51-1.53(m, 5H), 2.00(m, 2H), 2.47(m, 2H) 2.66(t, 1H), 3.06-3.17(m, 3H), 3.37(t, 1H), 3.56-3.70(m, 4H), 3.98( t, 1H), 4.14-4.27(m, 2H), 4.42(t, 1H), 4.82(q, 1H), 5.46(s, 2H), 7.05(q, 1H), 7.29(q, 1H), 7.52-7.69(m, 2H), 7.83(d, 1H), 8.13(d, 1H) JPEG2025539164000020.jpg37168

[0053] 3) Preparation of compound OZD-MET 01 Compound (f) and compound (d) were dissolved in DMF, and EDC, HOBT, and DIPEA were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure. The resulting mixture was separated using a silica gel column to obtain the white solid compound OZD-MET 01 in 75% yield.

[0054] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.51(d, 3H), 1.99(m, 2H), 2.45(m, 2H), 2.81(s, 3H) , 3.15-3.24(m, 4H), 3.45-3.59(m, 3H), 3.64-3.70(m, 3H), 3.98(t, 1H), 4.14(t, 1H), 4 .26-4.42(m, 3H), 4.73-4.74(m, 2H), 5.46(s, 2H), 6.41(s, 1H), 7.05-7.20(m, 2H), 7.4 0-7.47(m, 4H), 7.65-7.69(m, 2H), 7.83-7.97(m, 2H), 8.51-8.53(m, 2H), 10.07(s, 1H) JPEG2025539164000021.jpg43169 <Example 2> Preparation of 2-{2-[(4-{[1-(2,4-dihydroxy-5-isopropylphenyl)-N-methylformamido]methyl}phenyl)formamido]ethoxy}ethyl 4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}pyrazol-1-yl)piperidine-1-carboxylate (hereinafter referred to as "Compound OZD-MET 02") JPEG2025539164000022.jpg281361) Preparation of intermediate compound (i) (para) Methyl 4-formylbenzoate was dissolved in methanol, methylamine was added, and sodium borohydride was added and stirred at 0° C. After all the starting materials had reacted, the methanol was removed under reduced pressure, and the resulting mixture was dissolved in methylene chloride, washed with HO (brine), dried over MeSO, filtered, and the solvent was removed under reduced pressure to obtain the following compound (g).

[0055] Compound (g) and compound (b) from Example 1 were dissolved in dimethylformamide, EDC, HOBT, and DIPEA were added, and the reaction was terminated by stirring at room temperature for 12 hours. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure to obtain compound (e). Compound (h), a clear liquid, was obtained in 52% yield by separation using a silica gel column. JPEG2025539164000024.jpg47139

[0056] Compound (h) was dissolved in methanol:THF:HO (1:1:1), lithium hydroxide was added, and the mixture was allowed to react at room temperature for 15 hours. The solvent was removed, and the mixture was dissolved in HO. The mixture was then acidified with HCl, extracted with EA, washed with brine, and dried with MgSO to obtain foam solid compound (i) in nearly 100% yield. 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 2.81(s, 3H), 3.19(m, 1H), 4.42(t, 2H), 6.41(s, 1H), 7.42-7.61(m, 3H), 8.02(d, 2H), 10.07(s, 1H) JPEG2025539164000025.jpg45144

[0057] 2) Preparation of compound OZD-MET 02 Compound (i) and compound (f) prepared in Example 1, part 2) were dissolved in DMF, and EDC, EBOT, and DIPEA were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure to obtain the compound. The compound was then separated on a silica gel column to obtain the pale yellow solid compound OZD-MET 02 in a 44% yield.

[0058] 1H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.51(d, 3H), 1.99(m, 2H), 2.45(m, 2H), 2.81(s, 3H), 3.16-3.24(m, 4H), 3.45-3.59(m, 3H), 3.64-3.70(m, 3H), 3.98(t, 1H), 4.14(t, 1H), 4. 26-4.42(m, 3H), 4.71-4.74(m, 2H), 5.46(s, 2H), 6.41(s, 1H), 7.05-7.20(m, 2H), 7.42 -7.47(m, 4H), 7.58-7.65(m, 2H), 7.83(d, 2H), 8.05(d, 2H), 8.51(d, 1H), 10.07(s, 1H) Example 3 Preparation of N-{[4-({5-[4-(4-{6-amino-5-[1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-3-yl}pyrazol)-1-yl)piperidin-1-yl]-5-oxopentyl}carbamoyl)phenyl]methyl}-2,4-dihydroxy-5-isopropyl-N-methylbenzamide (hereinafter referred to as "Compound OZD-MET 03") JPEG2025539164000026.jpg331341) Preparation of Crizotinib-Linker Intermediate Compound (j) (R)-Crizotinib and 5-aminovaleric acid were dissolved in DMF, and EDC and HOBT were added to the solution. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with Brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure to give crude compound (j) in 28% yield.

[0059] 1 H NMR (500MHz, DMSO) δ1.36-1.39(m, 3H), 1.51-1.61(m, 5H), 1.84(m, 1H), 1.98-1.99(m, 3H), 2.40-2.53(m, 4H), 3.09(q, 2H), 3.53(0t, 2H), 4.29(m, 1H), 4.77(q, 1H), 5.46(s, 2H), 7.05(q, 1H), 7.30(q, 1H), 7.51-7.61(m, 2H), 7.83(d, 1H), 8.13(d, 1H) JPEG2025539164000027.jpg281392) Preparation of compound OZD-MET 03 Compound (j) and compound (i) prepared in Example 2, part 1) were dissolved in DMF, and EDC and HOBT were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure to obtain the compound. The compound was then purified by silica gel column separation to obtain yellow solid compound OZD-MET 03 in a 63% yield.

[0060] 1 H-NMR (500MHz, DMSO) δ1,24(d, 6H), 1.40(m, 1H), 1.51(d, 3H), 1.68(m, 2H), 1.90-1.98(m, 4H), 2.44-2.53(m, 3H), 2.81(s, 3H), 3.08(m, 2H), 3.13-3.19(m, 2H), 3.47-3.52(m, 3H), 4.28 -4.32(m, 2H), 4.68-4.75(m, 2H), 5.46(s, 2H), 6.41(s, 1H), 6.77(s, 1H), 7.05(q, 1H), 7. 21(q, 1H), 7.39-7.44(m, 4H), 7.65-7.83(m, 2H), 8.05(d, 2H), 8.51(d, 1H), 10.07(s, 1H) Example 4 Preparation of 2-{2-[(3-{[1-[2,4-dihydroxy-5-isopropylphenyl]-N-methylformamido]methyl}phenyl)formamido]ethoxy}ethyl 4-[3-(1-{imidazole[1,2-a]pyridin-6-yl}ethyl)-[1,2,3]triazolo[4,5-b]pyrazin-5-yl]pyrazole-1-carboxylate (hereinafter referred to as "Compound OZD-MET 04") JPEG2025539164000028.jpg321341) Preparation of savolitinib-linker intermediate compound (k) Compound (k), a savolitinib-linker intermediate having the following structure, was prepared in the same manner as in 2) of Example 1 above, using (R)-savolitinib instead of (R)-crizotinib (yield: 33%).

[0061] 1 H NMR (500MHz, DMSO) δ1.53(s, 2H), 2.11(d, 3H), 2.66(t, 1H)3.06(t, 1H), 3.37(t, 1H), 3.56(t, 1H), 3. 70(t, 1H), 3.98-4.14(m, 2H), 4.42(t, 1H), 6.01(m, 1H), 7.10(d, 1H), 8.00-8.25(m, 5H), 9.37(m, 2H) JPEG2025539164000029.jpg351212) Preparation of compound OZD-MET 04 Compound (k) and compound (d) prepared in Example 1, 1) were dissolved in DMF, and EDC, HOBT, and DIPEA were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure to obtain the compound. The solid compound OZD-MET 04 was obtained in 37% yield by silica gel column separation. 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 2.10(d, 3H), 2.81(s, 3H), 3.19(m, 1H), 3.41-3.45(m, 2H), 3.64-3.70(m, 2H), 3.98-4.14(m, 2H), 4.42-4.46(m, 2 H), 4.89(t, 1H), 6.00(m, 1H), 6.41(s, 1H), 7.40-7.42(m, 3H), 7.69-7.85 (m, 2H), 7.97-8.17(m, 5H), 8.57(m, 2H), 9.37-9.44(m, 2H), 10.07(s, 1H)

[0062] <Example 5> Preparation of 2,4-dihydroxy-N-({4-[(5-{4-[3-(1-{imidazole[1,2-a]]pyridin-6-yl}ethyl)-[1,2,3]triazolo[4,5-b]pyrazin-5-yl]pyrazol-1-yl}-5-oxopentyl)carbamoyl]phenyl}methyl)-5-isopropyl-N-methylbenzamide (hereinafter referred to as "Compound OZD-MET 05") JPEG2025539164000030.jpg361311) Preparation of savolitinib-linker intermediate compound (l) Compound (l), a savolitinib-linker intermediate having the following structure, was prepared in the same manner as in 1) of Example 3 above, except that (R)-savolitinib was used instead of (R)-crizotinib (yield: 39%).

[0063] 1 H NMR (500MHz, DMSO) δ1.36-1.39(m, 3H), 1.59(m, 1H), 1.91(m, 1H), 2.14-2.15(m, 4H), 2.40(t, 1H), 2.71 -2.79(m, 2H), 3.27(t, 1H), 6.05(m, 1H), 7.11(d, 1H), 7.96(d, 1H), 8.03-8.16(m, 4H), 9.33-9.37(m, 2H) JPEG2025539164000031.jpg601682) Preparation of compound OZD-MET 05 Compound (l) and compound (i) prepared in Example 2, part 1) were dissolved in DMF, and EDC and HOBT were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure to obtain the compound. The compound was then purified by silica gel column separation to obtain the white solid compound OZD-MET 05 in a 29% yield. 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.39(m, 1H), 1.70(m, 1H), 1.97-2.20(m, 5H), 2.71(t, 1H), 2.81(s, 3H), 3.13-3.27(m, 3H), 3.47(t, 1H), 4.48( t, 1H), 4.91(t, 1H), 6.03(m, 1H), 6.41(s, 1H), 6.77(s, 1H), 7.42-7.68( m, 4H), 8.02-8.17(m, 6H), 8.54(d, 1H), 9.37-9.39(m, 2H), 10.01(s, 1H)

[0064] Example 6 Preparation of 2-{2-[(3-{[1-(2,4-dihydroxy-5-isopropylphenyl)-N-methylformamido]methyl}phenyl)formamido]ethoxy}ethyl N-[4-(4-{1-[(4-fluorophenyl)carbamoyl]cyclopropanamido}phenoxy)quinolin-6-yl)carbamate (hereinafter referred to as "Compound OZD-MET 06") JPEG2025539164000032.jpg23140 1) Preparation of Cabozantinib-Linker Intermediate Compound (m) Compound (m), a cabozantinib-linker intermediate having the following structure, was prepared in the same manner as in 2) of Example 1 above, except that cabozantinib was used instead of R)-crizotinib (yield: 56%).

[0065] 1 H NMR (500MHz, DMSO) δ1.49-1.53(m, 4H), 1.72(q, 2H), 2.86(t, 2H), 3.46(t, 2H), 3.84(t, 2H), 4.28(t, 2H), 6.65(d, 1H), 6.95(d, 2H), 7.20-7.45(m, 5H), 7.71(d, 1H), 8.20(m, 3H), 8.59(s, 1H), 8.82(d, 1H), 9.83(s, 1H), 9.92(s, 1H) JPEG2025539164000033.jpg361142) Preparation of compound OZD-MET 06 Compound (m) and compound (d) prepared in Example 1, 1) were dissolved in DMF, and EDC and HOBT were added to the solution to react. NaHCO3 (aq.) was added to the solution, which was then extracted three times with EA, washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure to obtain the compound. The compound was then purified by silica gel column separation to obtain the dark yellow solid compound OZD-MET 06 in 42% yield.

[0066] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.48(q, 2H), 1.71(1, 2H), 2.81(s, 3H), 3.19-3.2 5(m, 3H), 3.55(5, 2H), 3.84(t, 2H), 4.28-4.32(m, 4H), 6.41(s, 1H), 6.65(d, 1H), 6 .95(d, 2H), 7.20-7.33(m, 4H), 7.40-7.45(m, 4H), 7.64-7.71(m, 2H), 7.97(m, 1H), 8.20-8.27(m, 3H), 8.44-8.59(m, 2H), 8.82(d, 1H), 9.43-9.52(m, 2H), 10.07(s, 1H) Example 7 Preparation of N'1-{4-[(6-{5-[(4-{[1-(2,4-dihydroxy-5-isopropylphenyl)-N-methylformamido]methyl}phenyl)formamido]pentanamido}quinolin-4-yl)oxy]phenyl}-N1-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide (hereinafter referred to as "Compound OZD-MET 07") JPEG2025539164000034.jpg371681) Preparation of Cabozantinib-Linker Intermediate Compound (n) Compound (n), a cabozantinib-linker intermediate having the following structure, was prepared in the same manner as in 1) of Example 3 above, except that cabozantinib was used instead of (R)-crizotinib (yield 63%).

[0067] 1 H NMR (500MHz, DMSO) δ1.39(s, 2H), 1.47-1.57(m, 6H), 1.76(q, 2H), 2.29(t, 2H), 2.59(t, 2H), 6.65(d, 1H), 6.95( d, 2H), 7.20-7.45(m, 5H), 7.71(d, 1H), 8.04-8.20(m, 3H), 8.49(s, 1H), 8.82(d, 1H), 10.05(s, 1H), 10.14(s, 1H) JPEG2025539164000035.jpg331305) Preparation of compound OZD-MET 07 Compound (n) and compound (i) prepared in Example 2, part 1) were dissolved in DMF, and EDC and HOBT were added to the solution to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure to obtain the compound. The solid compound OZD-MET 07 was obtained in a 54% yield by silica gel column separation.

[0068] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.51-1.54(m, 4H), 1.63-1.74(m, 4H), 2. 29(t, 2H), 2.81(s, 3H), 3.19-3.30(m, 3H), 4.34(t, 2H), 6.41(s, 1H), 6.65- 6.77(m, 2H), 6.95(d, 2H), 7.20(q, 2H), 7.33-7.45(m, 5H), 7.63-7.71(m, 2 H), 8.05-8.20(m, 6H), 8.82(d, 1H), 9.65(s, 1H), 9.75(s, 1H), 10.07(s, 1H) Example 8 Preparation of 2-{2-[(3-{[1-(2,4-dihydroxy-5-isopropylphenyl)-N-methylformamido]methyl}phenyl)formamido]ethoxy}ethyl N-{2-fluoro-4-[7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl]phenyl}carbamate (hereinafter referred to as "Compound OZD-MET 08") JPEG2025539164000036.jpg231281) Preparation of Capmatinib-Linker Intermediate Compound (o) Compound (o), a capmatinib-linker intermediate having the following structure, was prepared in the same manner as in 2) of Example 1 above, except that capmatinib was used instead of R)-crizotinib (yield: 58%).

[0069] 1 H NMR (500MHz, DMSO) δ1.53(s, 2H), 2.86(t, 2H), 3.46(t, 2H), 3.84(t, 2H), 4.24-4.28(m, 4H), 7 .19(m, 1H), 7.39-7.62(m, 4H), 7.91-8.04(m, 2H), 8.20-8.39(m, 3H), 8.87(q, 1H), 9.19(s, 1H) JPEG2025539164000037.jpg341202) Preparation of compound OZD-MET 08 Compound (o) and compound (d) prepared in Example 1, 1) were dissolved in DMF, and EDC and HOBT were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure to obtain the compound. The solid compound OZD-MET 08 was obtained in 55% yield by silica gel column separation.

[0070] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 2.81(s, 3H), 3.19(m, 1H), 3.46-3.55(m, 4H) , 3.84(t, 2H), 4.22-4.28(m, 4H), 4.53(t, 2H), 6.41(s, 1H), 7.19(m, 1H), 7.40 -7.42(m, 3H), 7.52-7.57(m, 2H), 7.62-7.72(m, 4H), 7.97(m, 1H), 8.12-8.21( m, 2H), 8.34(t, 1H), 8.44-8.46(m, 2H), 8.87(q, 1H), 9.01(s, 1H), 10.01(s, 1H) Example 9: Preparation of N-[(4-{[4-({2-fluoro-4-[7-(quinolin-6-ylmethyl)imidazo]1,2-b][1,2,4]triazin-2-yl]phenyl}carbamoyl)butyl]carbamoyl}phenyl)methyl]-2,4-dihydroxy-5-isopropyl-N-methyl-benzamide (hereinafter referred to as "compound OZD-MET 09") JPEG2025539164000038.jpg34154 1) Preparation of Capmatinib-Linker Intermediate Compound (p) Compound (p), a capmatinib-linker intermediate having the following structure, was prepared in the same manner as in 1) of Example 3 above, except that capmatinib was used instead of (R)-crizotinib (yield 60%).

[0071] 1 H NMR (500MHz, DMSO) δ1.39(s, 2H), 1.47(m, 2H), 1.77(m, 2H), 2.29(t, 2H), 2.59(t, 2H), 4.27(q, 2H), 7.19(m, 1H), 7.39-7.44(m, 3H), 7.62(d, 1H), 7.91(q, 1H), 8.21-8.40(m, 3H), 8.64(s, 1H), 8.87(q, 1H), 9.19(s, 1H) JPEG2025539164000039.jpg351182) Preparation of compound OZD-MET 09 Compound (p) and compound (i) prepared in Example 2, part 1) were dissolved in DMF, and EDC and HOBT were added to react. NaHCO3 (aq.) was added to the solution, which was then extracted three times with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure to obtain the compound. The compound was then separated using a silica gel column to obtain the compound OZD-MET 09 as a white solid in 63% yield.

[0072] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.54(m, 2H), 1.84(m, 2H), 2.29(t, 2H), 2.8 1(s, 3H), 3.19-3.30(m, 3H), 4.26(q, 2H), 4.55(t, 2H), 6.41(s, 1H), 6.77(s, 1H), 7.19(m, 1H), 7.42-7.44(m, 4H), 7.62-7.63(m, 2H), 8.05-8.12(m, 3H), 8 .21-8.25(m, 2H), 8.41-8.54(m, 2H), 8.87(q, 1H), 9.01(s, 1H), 10.07(s, 1H) Example 10 Preparation of 2-{2-[(3-{[1-(2,4-dihydroxy-5-isopropylphenyl)-N-methylformamido]methyl}phenyl)formamido]ethoxy}ethyl 4-({[2-(3-{[3-(3-cyanophenyl)-6-oxopyridazin-1-yl]methyl}phenyl)pyrimidin-5-yl]oxy}methyl)piperidine-1-carboxylate (hereinafter referred to as "Compound OZD-MET 10") JPEG2025539164000040.jpg29135 1) Preparation of Tepotinib-Linker Intermediate Compound (q) Compound (q), a tepotinib-linker intermediate having the following structure, was prepared in the same manner as in 2) of Example 1 above, except that tepotinib was used instead of R)-crizotinib (yield 76%).

[0073] 1 H NMR (500MHz, DMSO) δ1.53(s, 2H), 1.65(m, 2H), 2.04(m, 3H), 2.86(t, 2H), 3.05(q, 2H), 3.46-3.49(m, 4H), 3.84(t, 2H), 4 .07(d, 2H), 4.28(t, 2H), 5.64(t, 2H), 7.29-7.42(m, 4H), 7.70-7.77(m, 2H), 8.13-8.20(m, 3H), 8.63(s, 2H), 8.93(d, 1H) JPEG2025539164000041.jpg361242) Preparation of compound OZD-MET 10 Compound (q) and compound (d) prepared in Example 1, part 1) were dissolved in DMF, and EDC and HOBT were added to the solution to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent was removed under reduced pressure to obtain the compound. The compound was then purified by silica gel column separation to obtain white compound OZD-MET 10 in 78% yield.

[0074] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.63(m, 2H), 2.03(m, 3H), 2.81(s, 3H), 3.03- 3.19(m, 3H), 3.45-3.47(m, 4H), 3.55(t, 2H), 3.84(t, 2H), 4.08-4.28(m, 4H), 4. 53(t, 2H), 5.63(t, 2H), 6.41(s, 1H), 7.01(m, 1H), 7.29-7.42(m, 5H), 7.65-7.77 (m, 4H), 7.97(m, 1H), 8.20(m, 1H), 8.34-8.45(m, 3H), 8.63(s, 2H), 10.07(s, 1H) Example 11 Preparation of N-{[4-({5-[4-({[2-(3-{[3-(3-cyanophenyl)-6-oxopyridazin-1-yl]methyl}phenyl)pyrimidin-5-yl]oxy}methyl)piperidin-1-yl]-5-oxopentyl}carbamoyl)phenyl]methyl}-2,4-dihydroxy-5-isopropyl-N-methylbenzamide (hereinafter referred to as "Compound OZD-MET 11") JPEG2025539164000042.jpg36166 1) Preparation of Tepotinib-Linker Intermediate Compound (r) Compound (r), a tepotinib-linker intermediate having the following structure, was prepared in the same manner as in 1) of Example 3 above, except that tepotinib was used instead of (R)-crizotinib (yield 74%).

[0075] 1 H NMR (500MHz, DMSO) δ1.39(s, 2H), 1.47(m, 2H), 1.63-1.74(m, 4H), 2.03-2.26(m, 5H), 2.59(t, 2H), 2.97(q, 2H), 3.41 (t, 2H), 4.07(d, 2H), 5.68(t, 2H), 7.29-7.42(m, 4H), 7.70-7.77(m, 2H), 8.13-8.20(m, 3H), 8.63(s, 2H), 8.93(d, 1H) JPEG2025539164000043.jpg321222) Preparation of compound OZD-MET 11 Compound (r) and compound (i) prepared in Example 2, part 1) were dissolved in DMF, and EDC and HOBT were added to the solution to react. NaHCO3 (aq.) was added to the solution, which was then extracted twice with EA, washed with brine, dried over MgSO4, filtered, and the solvent removed under reduced pressure to obtain the compound. The compound was then purified by silica gel column separation to obtain OZD-MET 11 as a yellow solid in 79% yield.

[0076] 1 H NMR (500MHz, DMSO) δ1.24(d, 6H), 1.54-1.62(m, 4H), 1.80(m, 2H), 2.01-2.05(m , 3H), 2.26(t, 2H), 2.96(q, 2H), 3.19(m, 1H), 3.30-3.40(m, 4H), 4.08(d, 2H), 4. 55(t, 2H), 5.67(t, 2H), 6.41(s, 1H), 6.77(s, 1H), 7.01(m, 1H), 7.29-7.42(m, 5H) ), 7.65-7.77(m, 3H), 8.05(d, 2H), 8.20-8.40(m, 3H), 9.43(d, 1H), 10.07(s, 1H) <Experimental Example 1> Cell viability effect This experiment was conducted to confirm the cytotoxicity and changes in cell number after treating cells with each of the compounds prepared in Examples 1 to 11. The IC 50 The cancer cell growth inhibitory and anti-cancer effects of the compounds of the present invention can be evaluated based on the values.

[0077] <Experimental Method> Cell viability confirmation test In a 96-well plate, 3 × 10 cells (H596 or H1437 cells) were placed in each well. 3After seeding the cells, they are cultured for 12 hours or more. After removing the medium, each compound prepared in Examples 1 to 11 is diluted and dispensed in 200 μl portions into the wells. After 3 days, the medium is removed, and 10 μl of CCK8 reagent + 90 μl of medium is added to each well. After 3 to 4 hours of incubation, the absorbance is measured using an ELISA reader at a wavelength of 450 nM. IC was calculated using the Graphpad Prism program. 50 The values ​​were derived and are shown in Table 2.

[0078] As can be seen from Table 2, the cell viability of each of the compounds prepared in Examples 1 to 11 was approximately 10 times higher than that of the control drug crizotinib, intermediate compound (d) of the first moiety that binds to the chaperone complex, and compound (i).

[0079] Therefore, the compound of the present invention has a much superior anti-cancer effect compared to the compound having the first moiety and the second moiety.

[0080] [Table 2]

[0081] <Experimental Example 2> Confirmation of c-MET degradation effect In this experiment, cells were treated with each of the compounds prepared in Examples 1 to 11, and the tendency and degree of degradation of the target protein c-MET (total form) itself in the cells was evaluated.

[0082] <Experimental Method> Test to confirm degradation of target protein 5 x 10 cells (H596 or H1437 cells) in a Φ60 culture dish or Φ100 culture dish 5 or 1 x 10 6After seeding, the cells were cultured for at least 12 hours. After removing the medium, the cells were treated with each of the compounds prepared in Examples 1-11 according to the conditions, i.e., concentration (0, 2.5, 5, 10 μM), time (0, 6, 8, 24, 48, 72 hours), or the same constant concentration and time (5 μM, 72 hours) for all compounds. Cells were then harvested and lysed using RIPA buffer. After centrifugation (4°C, 13,000 rpm, 15 min), the supernatant was transferred to a new tube. Protein was quantified using a Bradford assay, and the loading sample was prepared by boiling at 95°C and subjected to Western blotting. The sample was loaded onto an 8% SDS polyacrylamide gel and electrophoresed at 80 V, then switched to 100–120 V (when the loaded sample migrated from the stacking gel to the running gel). The electrophoresed polyacrylamide gel was transferred to a membrane (transfer buffer was diluted with 10X transfer buffer:methanol:DW = 1:2:7, and then incubated at 100 V for 1 hour or 80 V for 2 hours in cold conditions). The membrane was then blocked with 5% skim milk for 30 minutes and washed with PBS-T (phosphate-buffered saline with Tween 20). The membrane was incubated with a primary antibody diluted in 5% BSA + NaN3 solution for at least 12 hours, then washed with PBS-T and incubated with 5% skim milk + secondary antibody for 1 hour and 30 minutes. After washing with PBS-T, the membrane was reacted with ECL solution, and the bands were detected using an imaging device to confirm the degradation of the band corresponding to the target protein c-MET. The results are shown in Table 3 and Figures 1 to 6.

[0083] As can be seen from Table 3 below, crizotinib, compound (d) having a first moiety that binds to the chaperone complex, and compound (i) did not show any degradation of the target MET protein, whereas each of the compounds prepared in Examples 1 to 11 clearly showed degradation of the target protein.

[0084] Therefore, the present invention has an extremely excellent decomposition efficiency for the target protein.

[0085] [Table 3] <Experimental Example 3> Target protein degradation and recovery by the UPS pathway In this experiment, each of the compounds prepared in Examples 1 to 11 was treated simultaneously with the proteasome inhibitor bortezomib to evaluate the degradation and recovery of target proteins via the ubiquitin-proteasome system (UPS) pathway.

[0086] <Experimental Method> Target protein degradation and recovery confirmation test 3 x 10 H596 or H1437 cells were cultured in a 6-well culture plate. 5After seeding, the cells were cultured for at least 12 hours. After changing the medium, 100 nM bortezomib and 5 μM of each compound prepared in Examples 1 to 11 were treated together. A control group was treated with the same volume of dimethylsulfoxide (DMSO), the drug solvent. After 16 hours, the medium was removed, the cells were washed with phosphate-buffered saline (PBS), and harvested using trypsin-ethylenediaminetetraacetic acid (Trypsin-EDTA). Cell lysis was performed with RIPA buffer (Radio-ImmunoPrecipitation Assay buffer), centrifuged (4°C, 13,000 rpm, 15 min), and the supernatant was transferred to a new tube. Protein quantification was performed using the Bradford assay, followed by boiling at 95°C for 10 minutes. Western blotting is performed on an 8% SDS polyacrylamide gel (sodium dodecyl sulfate polyacrylamide gel) at 80V. Once proteins have migrated from the stacking gel to the running gel, the voltage is changed to 100V and the gel is left cold for 60 minutes. The gel is then blocked with 5% skim milk for at least 30 minutes and washed with PBS-T (phosphate-buffered saline with Tween 20). The primary antibody is diluted in 5% bovine serum albumin (5% BSA) and NaN3 solution and incubated for at least 12 hours. After washing with PBS-T, the secondary antibody is diluted in 5% skim milk and incubated for 1 hour and 30 minutes. After washing with PBS-T, the membrane is incubated with ECL solution and the bands are confirmed using an imaging device. The results are shown in Table 4 and Figures 7 to 12 to confirm whether the reduction in MET expression caused by each of the compounds prepared in Examples 1 to 11 was restored when treated with bortezomib.

[0087] As can be seen from Table 4, crizotinib, intermediate compound (d) of the first moiety that binds to the chaperone complex, and compound (i) did not show degradation or recovery of the target MET protein. On the other hand, each of the compounds prepared in Examples 1 to 11 clearly showed degradation of the target protein and recovery of the target protein upon treatment with the proteasome inhibitor bortezomib.

[0088] [Table 4]

Claims

1. A compound represented by the following chemical formula I or a stereoisomer thereof. <Chemical formula I> CB-L-TB (In the formula, CB is and TB is is one selected from L is It is one of the following:

2. 2. The compound of formula I or a stereoisomer thereof according to claim 1, wherein CB is present in meta or para form.

3. TB is 2. The compound of claim 1, wherein the compound is selected from the group consisting of:

4. L 2. The compound of formula I according to claim 1, wherein:

5. A compound of Formula I selected from the group consisting of:

6. A pharmaceutical composition for treating cancer disease, comprising the compound of Chemical Formula 1 according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

7. The cancer disease is selected from the group consisting of acoustic neurinoma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia (monocytic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic), acute T-cell leukemia, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchioloalveolar carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, chronic myeloid leukemia, colorectal cancer, colon cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and the like. lymphoma), dysproliferative changes (dysplasia and metaplasia), embryonal carcinoma, endometrial carcinoma, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal carcinoma, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor tumor), fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, heavy chain disease, hemangioblastoma, liver tumor, hepatocellular carcinoma, hormone-refractory prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin's lymphoma and non-Hodgkin's lymphoma), malignant tumors and hyperproliferative disorders of the bladder, breast, colon, lung, ovary, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma , mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung carcinoma, solid tumors (carcinoma and sarcoma), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovium, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular tumor, uterine cancer, Wilms' tumor tumor), primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelial cancer, gynecological cancer, uterine cancer, gestational trophoblastic disease, male reproductive organ cancer, seminal vesicle cancer, testicular cancer, germ cell tumor, endocrine tumor, thyroid cancer, adrenal cancer, pituitary cancer, hemangioma, sarcoma derived from bone and soft tissue, Kaposi's sarcomasarcoma), nerve cancer, eye cancer, meningeal cancer, glioblastoma, neuroma, neuroblastoma, Schwannomas, hematopoietic malignancies, such as solid tumors resulting from leukemia, metastatic melanoma, recurrent or persistent epithelial ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumor, colon cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small cell lung cancer, malignant glioma, epithelial ovarian cancer, primary peritoneal serous carcinoma, metastatic liver cancer, neuroendocrine cancer, refractory malignancies, triple-negative breast cancer, HER2-amplified breast cancer, nasopharyngeal carcinoma, oral cancer, bile duct cancer, hepatocellular carcinoma 10. The pharmaceutical composition for treating cancer diseases according to claim 6, wherein the cancer is selected from the group consisting of squamous cell carcinoma of the head and neck (SCCHN), non-medullary thyroid cancer, recurrent glioblastoma multiforme, neurofibromatosis type 1, central nervous system cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumor, triple-negative breast cancer, colon cancer, sarcoma, melanoma, kidney cancer, endometrial cancer, thyroid cancer, rhabdomyosarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumor, mantle cell lymphoma, and refractory malignant tumors.

Images