Novel hemiastalin derivatives and their applications

Abstract

This invention provides a novel hemiastalin derivative, a method for producing the same, and its applications. In this invention, the anticancer effect is confirmed using a novel hemiastalin derivative with improved stability.

Description

Technical Field

[0001] The present invention provides a novel hemiasterlin derivative, a method for producing the same, and uses thereof. The novel hemiasterlin derivative synthesized in the present invention exhibits a significant survival inhibitory effect against cancer cell lines and can be used as a safe form of anticancer agent.

Background Art

[0002] Hemiasterlin can be isolated from marine sponges of the genus Cymbastela sp., Hemiasterella minor, Siphonochalina sp., and Auletta sp. (Talpir et al., Tetrahedron Letters, vol. 35, no. 25, pp. 4453-4456, 1994).

[0003] Natural hemiasterlin is known as a drug that suppresses the formation of microtubules, which are essential for cancer cell division, and has been actively studied for the treatment of cancers such as ovarian cancer. However, safety problems still exist.

[0004] Thereafter, the present inventors completed the present invention by confirming the structure of a derivative of hemiasterlin with improved safety as a therapeutic drug and confirming an anticancer effect similar to that of natural hemiasterlin.

Summary of the Invention

Problems to be Solved by the Invention

[0005] In order to solve the above-mentioned problems, the present invention aims to confirm the structure of a novel hemiasterlin derivative with improved safety and use it as a drug for cancer prevention or treatment, and provide it in the form of a drug-linker, a carrier drug conjugate (ADC), etc.

Means for Solving the Problems

[0006] To achieve the above objective, the present invention provides a novel hemiasterlin derivative, the compound of the following chemical formula 1, its isomer, its pharmaceutically acceptable salt, or its solvate. [Chemical formula 1] [ka] In the above chemical formula 1, R is selected from halogen, hydroxyl, carboxyl group, C1-C6 alkyl, and C1-C6 alkoxy.

[0007] In the present invention, derivatives or stereoisomers of the compound of chemical formula 1 include, but are not limited to, the compounds represented by the following chemical formulas 2 to 7. [Chemical formula 2] [ka] [Chemical formula 3] [ka] [Chemical formula 4] [ka] [Chemical formula 5] [ka] [Chemical formula 6] [ka] [Chemical formula 7] [ka]

[0008] The present invention also provides a “carrier-drug complex” or “carrier-linker-drug complex” comprising a compound of chemical formula 1, its derivatives, its isomers, its pharmaceutically acceptable salts or solvates; and a carrier linked to the compound.

[0009] Other embodiments of the present invention provide a “drug-linker” comprising a compound of chemical formula 1, its derivatives, its isomers, its pharmaceutically acceptable salts or solvates; and a linker linked to the compound.

[0010] In the present invention, the linker includes, but is not limited to, GGFG or val-cit, and any linker commonly used in the industry can be used without restriction.

[0011] In the present invention, "Carrier" means a substance that has the ability to selectively and specifically deliver the compound according to the present invention to a target site, such as cancer cells, and may be, but is not limited to, an antibody, peptide, repetibody, and / or aptamer, but is preferably an antibody. For example, the carrier of the present invention may be 4-1BB, 5T4, integrin, activin, amyloid beta, angiopoietin (angiopoietin 1 or 2), angiopoietin-like substance 3, B cell maturation antigen (BCMA), B cell activating factor (BAFF), B7-H3, complement 5. 5), CCR4, CCR5, CCL11, CD2, CD3, CD4, CD6, CD11a, CD16A, CD19, CD20, CD22, CD25, CD27, CD28, CD30, CD32B, CD33, CD38, CD40, CD45, CD46, CD 47, CD52, CD56, CD62, CD70, CD73, CD74, CD79b, CD80, CD105, CD123, CD154, CD166, CD262, CD278, CD319, CD326, Carcinoembryonic antigen, CEA), CGRP, Claudin-18, c-Met, CSF-1, CSF-1 receptor, CTLA4, DLL3, EGF receptor, hemophilia factor, Fc receptor, FGF23, folate receptor, GD2, Glucocorticoid-induced TNF receptor (GITR), Glypican 3, GM-CSF, HER2, HER3, TROP2, Hepatocyte Growth FactorFactor (HGF), interferon receptor, interferon gamma, IgE, IGF-1 receptor, interleukin 1, interleukin 2 receptor, interleukin 4, interleukin 4 receptor, interleukin 5, interleukin 5 receptor, interleukin 6, interleukin 6 receptor, interleukin 8, interleukin 12 / 23, interleukin 13, interleukin 17A, interleukin 17 receptor A, interleukin 23, interleukin 31 receptor, interleukin 36 receptor, lymphocyte-activation gene 3 (LAG3), lysyl oxidase homolog 2 (LOXL2), mesothelin, mucin-1, mucin-16, netin-4, nerve growth factor Factor (NGF), OX40, Proprotein Convertase Subtilisin / Kexin type 9 (PCSK9), PD-1, PD-L1, Phospholipase C, RANKL (Receptor activator of nuclear factors kappa B ligand), Tyrosine-protein kinase transmembrane receptor (ROR1), Sialic acid binding ig-like lectin 15 (Siglec-15), Transforming growth factor beta (TGFβ), TIGIT (T-cell innunoreceptor with immunoglobulin and ITIM domain), T cell immunoglobulin and mucin-domain containing-3 (Tim-3), Tissue factor, Tissue factor pathway inhibitor (Tissue factorIt is an antibody, peptide, repebody, or aptamer that specifically binds to one or more substances (antigens) selected from the group consisting of pathway inhibitors (TFPI), TORP-2, tumor necrosis factor (TNF), thymic stromal lymphopoietin (TSLB), colony-stimulating factor 1 receptor (CSF1R), vascular endothelial growth factor (VEGF), VEGF receptor, and vWF (von Willebrand Factor).

[0012] For example, in the present invention, the antibodies are urelumab, utomilumab, bebtelovimab, aducanumab, bapineuzumab, crenezumab, donanemab, gantenerumab, lecanemab, solanezumab, nesvacumab, and evinacumab. ), Enoblituzumab, Omburtamab, Belimumab, Ianalumab, Tabalumab, Bertilimumab, Mogamulizumab, Leronlimab, Siplizumab, Foralumab, Muromonab-CD3, Otelixizumab, Tep Teplizumab, Ibalizumab, Tregalizumab, Zanolimumab, Itolizumab, Efalizumab, Inebilizumab, Tafasitamab, Tositumomab, Ocrelizumab, Ofatumumab, Rituximab, Ubrituximab Ublituximab, Veltuzumab, Epratuzumab, Basiliximab, Daclizumab, Varlilumab, Lulizumab, Iratumumab, Lintuzumab, Daratumumab, Felzartamab, Isatuximab, Mezagitamab,Bleserumab, Dacetuzumab, Iscalimab, Lucatumumab, Mitazalimab, Sotigalimab, Dapirolizumab, Apamistamab, Ligufalimab, Magrolimab, Alemtuzumab, Crizanlizumab, In Inclacumab, Cusatuzumab, Oleclumab, Miratuzumab, Galiximab, Carotuximab, Adecatumumab, Eptinezumab, Erenumab, Fremanezumab, Galcanezumab, Zolbetuximab, Onartuzumab nartuzumab, eculizumab, pozelimab, ravolizumab, lacnotuzumab, axatilimab, cabiralizumab, emactuzumab, ipilimumab, quavonlimab, tremelimumab, zalifrelimab, cetuximab imab), Depatuxizumab, Futuximab, Imgatuzumab, Matuzumab, Modotuximab, Necitumumab, Nimotuzumab, Panitumumab, Tomuzotuximab, Zalutumumab, Batoclimab, Nipocalimab,Rozanolixizumab, Burosumab, Farletuzumab, Dinutuximab, Naxitamab, Ragifilimab, Gimsilumab, Lenzilumab, Mavrilimumab, Namilumab, Otilimab, Plonmarlimab, Kodori Codrituzumab, Margetuximab, Pertuzumab, Trastuzumab, Datopotamab, Patritumab, Seribantumab, Duligotuzumab, Ficlatuzumab, Rilotumumab, Alomfilimab, Aniflorumab olumab), Emapalumab, Ligelizumab, Omalizumab, Cixutumumab, Dalotuzumab, Figitumumab, Ganitumab, Teprotumumab, Bermekimab, Canakinumab, Gevokizumab, Briakinumab ), ustekinumab, anrukinzumab, sendakimab, lebrikizumab, tralokinumab, brodalumab, bimekizumab, ixekizumab, secukinumab, brazikumab, guselkumab, mirikizumab,Risankizumab, Tildrakizumab, Nemolizumab, Imsidolimab, Spesolimab, Pascolizumab, Dupilumab, Depemokimab, Mepolizumab, Reslizumab, Benralizumab, Clazakizumab Olokizumab, Siltuximab, Sirukumab, Ziltivekimab, Levirimab, Sarilumab, Satralizumab, Tocilizumab, Abituzumab, Favezelimab, Fianlimab, Ieramilimab, Relamilimab (atlimab), Simtuzumab, Abagobomab, Oregovomab, Tanezumab, Ivuxolimab, Rocatinlimab, Tavolimab, Telazorlimab, Vonlerolizumab, Alirocumab, Bococizumab, Ebronucimab ), Evolocumab, Frovocimab, Ongericimab, Tafolecimab, Dostarlimab, Balstilimab, Camrelizumab, Cemiplimab, Geptanolimab, Nivolumab, Pembrolizumab, Penpulimab,Pidilizumab, Prolgolimab, Retifanlimab, Sasanlimab, Serplulimab, Sintilimab, Spartalizumab, Tislelizumab, Toripalimab, Ezabenlimab, Zimberelimab, Atezolizumab Lizumab), Avelumab, Cosibelimab, Sugemalimab, Durvalumab, Envafolimab, Suvratoxumab, Denosumab, Zilovertamab, Elotuzumab, Domvanalimab, Etigilimab, Ociperlima b) Tiragolumab, Vibostolimab, Surzebiclimab, Cobolimab, Sabatolimab, Concizumab, Marstacimab, Adalimumab, Golimumab, Infliximab, Certolizumab, Conatumumab, Gatuzumab, Tezepelumab, Gatipotuzumab, Cabiralizumab, Bevacizumab, Brolucizumab, Ranibizumab, Olinvacimab, Icrucumab, Ramucirumab, Caplacizumab, Abrilumab,Etrolizumab, Vedolizumab, Intetumumab, Natalizumab, Obrindatamab, Elranatamab, Linvoseltamab, Teclistamab, Epco, Epcoritamab, Glofitamab, Mosunetuzumab, Odronextamab, Flotetuzumab, Vibecotamab, Catumaxomab, Cibisatamab, Talquetamab, Ubamatamab, Emfizatamab, Blinatumomab ab) Amivantamab, Emicizumab, Zenocutuzumab, Zanidatamab, Tibulizumab, Naptumomab, Belantamab, Pivekimab, Praluzatamab, Coltuximab, Denintuzumab, Loncastuximab, Ibritumomab, Inotuzumab, Epratuzumab, Moxetumomab, Brentuximab, Gemtuzumab, Vadastuximab, Lorvotuzumab, Polatuzumab, Tusamitamab, Telisotuzumab, Rovalpituzumab zumab), Depatuxizumab, Farletuzumab, Mirvetuximab, Disitamab, Anetumab, Enfortumab, Sacituzumab, Vobarilizumab, Cadonilimab, Vudalimab, Tebotelimbab, Ivonescimab,This includes, but is not limited to, erfonrilimab, ozoralizumab, faricimab, vanucizumab, and navicixizumab.

[0013] The present invention also provides a "pharmaceutical composition for the prevention or treatment of cancer" comprising a compound of chemical formula 1, its isomers, a pharmaceutically acceptable salt thereof, or a solvate thereof, and "cancer prevention or treatment uses" of the compound represented by chemical formula 1.

[0014] Here, the compound of chemical formula 1 can be provided in a form connected to a carrier via a linker.

[0015] In the present invention, the cancers include pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, epithelial ovarian cancer, ovarian germ cell tumor, male breast cancer, brain tumor, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, nasal cavity cancer / paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain tumor, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, Renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue tumors, malignant bone tumors, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary origin, gastric lymphoma, stomach cancer, gastric carcinoid tumor, gastrointestinal stromal tumor, Wilms' tumor, breast cancer, triple-negative breast cancer (TNBC), sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, intrauterine cancer This includes, but is not limited to, membrane cancer, uterine sarcoma, prostate cancer, metastatic bone tumors, metastatic brain tumors, mediastinal tumors, rectal cancer, rectal carcinoid tumors, vaginal cancer, spinal cord cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, hematological cancer, and thymic cancer. [Effects of the Invention]

[0016] This invention provides novel hemiastalin derivatives and methods for producing the same. The novel hemiastalin derivatives according to the present invention can be used as safe and effective pharmaceutical compositions. [Brief explanation of the drawing]

[0017] [Figure 1] Figure 1 shows the results of measuring cell viability at different treatment concentrations after treating lung cancer cell lines (A549) with the compound of chemical formula 2, SC209, and MMAE, followed by 3 days of incubative incubation. [Figure 2]Figure 2 shows the results of measuring cell viability at different treatment concentrations after treating head and neck squamous cell carcinoma cell lines (FaDu) with the compound of chemical formula 2, SC209, and MMAE, followed by 3 days of incubative incubation. [Best Mode for Carrying Out the Invention]

[0018] The following are various embodiments to aid in understanding the invention. These embodiments are provided solely to facilitate understanding of the invention, and the scope of protection of the invention is not limited to these embodiments.

[0019] The present invention provides a compound of chemical formula 1 as a derivative of hemiastalin, its isomers, its pharmaceutically acceptable salts or solvates. [Chemical formula 1] [ka] In the above chemical formula 1, R is selected from halogen, hydroxyl, carboxyl group, C1-C6 alkyl, and C1-C6 alkoxy.

[0020] In this specification, hemiasterlin is defined as C 30 H 46 It is a compound with the molecular formula N4O4 (molecular weight: 526.7 g / mol) and has the following chemical structure. [Hemiastarin] [ka]

[0021] In this specification, “isomer” means a relationship between compounds that have the same chemical formula but do not have identical properties. Such types of isomers include structural isomers and stereoisomers. “Stereoisomer” means a compound that has the same chemical composition but differs in terms of the arrangement of atoms or groups in space. This includes not only optical isomers (e.g., essentially pure enantiomers, essentially pure diastereomers, or mixtures thereof), but also conformation isomers (i.e., isomers that differ only in the angle of one or more chemical bonds), position isomers (in particular tautomers), or geometric isomers (e.g., cis-trans isomers). In this specification, isomers of a compound of chemical formula 1 mean stereoisomers, but are not limited to stereoisomers.

[0022] The compound of chemical formula 1 according to the present invention is specifically selected from the following compounds and may be in the form of each isomer compound or a mixture thereof. [ka]

[0023] In this specification, "salt" means a salt according to one embodiment of the present disclosure that has the preferred pharmacological activity of the parent compound, and can be a salt that does not cause severe irritation to the organism to which the compound is applied and does not impair the biological activity and physical properties of the compound. Examples include salts of inorganic ions such as sodium, potassium, calcium, magnesium, lithium, copper, manganese, zinc, and iron, and salts of inorganic acids such as hydrochloric acid, phosphoric acid, and sulfuric acid. Other examples include salts of organic acids such as ascorbic acid, citric acid, tartaric acid, lactic acid, maleic acid, malonic acid, fumaric acid, glycolic acid, succinic acid, propionic acid, acetic acid, orotic acid, and acetylsalicylic acid, as well as amino acid salts such as lysine, arginine, and guanidine.

[0024] Where the composition described herein is a pharmaceutical composition, the “salt” may be a “pharmaceutically acceptable salt.” “pharmaceutically acceptable” means that it can be approved, has been approved, is listed in a pharmacopoeia, or is recognized as such in other common pharmacopoeias, for use in animals, more specifically in humans, when used in a normal medicinal dosage, while avoiding significant toxic effects.

[0025] In this specification, “salt” or “pharmaceutically acceptable salt” means an inorganic salt, organic salt, or addition salt of a metal salt of a compound. The inorganic salt may be a hydrochloride, bromate, phosphate, sulfate, or disulfate. The organic salt may be a formate, acetate, propionate, lactate, oxalate, tartrate, malate, maleate, citrate, fumarate, besylate, camusylate, edisylate, trichloroacetate, trifluoroacetate, benzoate, gluconate, methanesulfonate, glycolate, succinate, 4-toluenesulfonate, galacturonate, embonate, glutamate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, or aspartate. The metal salt may be a calcium salt, sodium salt, magnesium salt, strontium salt, or potassium salt.

[0026] In this specification, "solvate" means a compound solvated in an organic or inorganic solvent. In this specification, such solvate is, for example, a hydrate.

[0027] The compound of the present invention (Chemical Formula 1) can be provided in a form connected to a carrier.

[0028] In the present invention, "Carrier" means a substance that has the ability to selectively and specifically deliver the compound according to the present invention to a target site, such as cancer cells, and may be, but is not limited to, an antibody, peptide, repel body, and / or aptamer, but is preferably an antibody.

[0029] For example, the carrier of the present invention may contain 4-1BB, 5T4, integrin, activin, amyloid beta, angiopoietin (angiopoietin 1 or 2), angiopoietin-like substance 3, B cell maturation antigen (BCMA), B cell activating factor (BAFF), B7-H3, and complement 5. 5), CCR4, CCR5, CCL11, CD2, CD3, CD4, CD6, CD11a, CD16A, CD19, CD20, CD22, CD25, CD27, CD28, CD30, CD32B, CD33, CD38, CD40, CD45, CD46, CD 47, CD52, CD56, CD62, CD70, CD73, CD74, CD79b, CD80, CD105, CD123, CD154, CD166, CD262, CD278, CD319, CD326, Carcinoembryonic antigen, CEA), CGRP, Claudin-18, c-Met, CSF-1, CSF-1 receptor, CTLA4, DLL3, EGF receptor, hemophilia factor, Fc receptor, FGF23, folate receptor, GD2, Glucocorticoid-induced TNF receptor (GITR), Glypican 3, GM-CSF, HER2, HER3, TROP2, Hepatocyte Growth Factor Factor (HGF), interferon receptor, interferon gamma, IgE, IGF-1 receptor, interleukin 1, interleukin 2 receptor, interleukin 4, interleukin 4 receptor, interleukin 5, interleukin 5 receptor, interleukin 6, interleukin 6 receptor, interleukin 8, interleukin 12 / 23, interleukin 13, interleukin 17A, interleukin 17 receptor A, interleukin 23, interleukin 31 receptor, interleukin 36 receptor, lymphocyte-activation gene 33, LAG3), Lysyl oxidase homolog 2 (LOXL2), Mesothelin, Mucin-1, Mucin-16, Netin-4, Nerve Growth Factor (NGF), OX40, Proprotein Convertase Subtilisin / Kexin type 9 (PCSK9), PD-1, PD-L1, Phospholipase C, RANKL (Receptor activator of nuclear factors kappa B ligand), Tyrosine-protein kinase transmembrane receptor (ROR1), Sialic acid-binding ig-like lectin 15 15, Siglec-15), Transforming growth factor beta (TGFβ), TIGIT (T-cell innunoreceptor with immunoglobulin and ITIM domain), T cell immunoglobulin and mucin-domain containing-3 (Tim-3), Tissue factor, Tissue factor pathway inhibitor (TFPI), TORP-2, Tumor necrosis factor (TNF), Thymic stromal lymphopoietin (TSLB), Colony stimulating factor 1 receptor (CSF1R), Vascular endothelial growth factor (VEGF), VEGF receptor and vWF (von WillebrandAn antibody, peptide, repebody, or aptamer that specifically binds to one or more substances (antigens) selected from the group consisting of Factors.

[0030] In the present invention, non-limiting examples of antibodies include urelumab, utomilumab, bebtelovimab, aducanumab, bapineuzumab, crenezumab, donanemab, gantenerumab, lecanemab, solanezumab, nesvacumab, and evinacumub. mab), Enoblituzumab, Omburtamab, Belimumab, Ianalumab, Tabalumab, Bertilimumab, Mogamulizumab, Leronlimab, Siplizumab, Foralumab, Muromonab-CD3, Otelixizumab, Teplizumab, Ibalizumab, Tregalizumab, Zanolimumab, Itolizumab, Efalizumab, Inebilizumab, Tafasitamab, Tositumomab, Ocrelizumab, Ofatumumab, Rituximab, Ubrituximab (Ublituximab), Veltuzumab, Epratuzumab, Basiliximab, Daclizumab, Varlilumab, Lulizumab, Iratumumab, Lintuzumab, Daratumumab, Felzartamab, Isatuximab, Mezagitamab,Bleserumab, Dacetuzumab, Iscalimab, Lucatumumab, Mitazalimab, Sotigalimab, Dapirolizumab, Apamistamab, Ligufalimab, Magrolimab, Alemtuzumab, Crizanlizumab, In Inclacumab, Cusatuzumab, Oleclumab, Miratuzumab, Galiximab, Carotuximab, Adecatumumab, Eptinezumab, Erenumab, Fremanezumab, Galcanezumab, Zolbetuximab, Onartuzumab nartuzumab, eculizumab, pozelimab, ravolizumab, lacnotuzumab, axatilimab, cabiralizumab, emactuzumab, ipilimumab, quavonlimab, tremelimumab, zalifrelimab, cetuximab imab), Depatuxizumab, Futuximab, Imgatuzumab, Matuzumab, Modotuximab, Necitumumab, Nimotuzumab, Panitumumab, Tomuzotuximab, Zalutumumab, Batoclimab, Nipocalimab,Rozanolixizumab, Burosumab, Farletuzumab, Dinutuximab, Naxitamab, Ragifilimab, Gimsilumab, Lenzilumab, Mavrilimumab, Namilumab, Otilimab, Plonmarlimab, Kodori Codrituzumab, Margetuximab, Pertuzumab, Trastuzumab, Datopotamab, Patritumab, Seribantumab, Duligotuzumab, Ficlatuzumab, Rilotumumab, Alomfilimab, Aniflorumab olumab), Emapalumab, Ligelizumab, Omalizumab, Cixutumumab, Dalotuzumab, Figitumumab, Ganitumab, Teprotumumab, Bermekimab, Canakinumab, Gevokizumab, Briakinumab ), ustekinumab, anrukinzumab, sendakimab, lebrikizumab, tralokinumab, brodalumab, bimekizumab, ixekizumab, secukinumab, brazikumab, guselkumab, mirikizumab,Risankizumab, Tildrakizumab, Nemolizumab, Imsidolimab, Spesolimab, Pascolizumab, Dupilumab, Depemokimab, Mepolizumab, Reslizumab, Benralizumab, Clazakizumab Olokizumab, Siltuximab, Sirukumab, Ziltivekimab, Levirimab, Sarilumab, Satralizumab, Tocilizumab, Abituzumab, Favezelimab, Fianlimab, Ieramilimab, Relamilimab (atlimab), Simtuzumab, Abagobomab, Oregovomab, Tanezumab, Ivuxolimab, Rocatinlimab, Tavolimab, Telazorlimab, Vonlerolizumab, Alirocumab, Bococizumab, Ebronucimab ), Evolocumab, Frovocimab, Ongericimab, Tafolecimab, Dostarlimab, Balstilimab, Camrelizumab, Cemiplimab, Geptanolimab, Nivolumab, Pembrolizumab, Penpulimab,Pidilizumab, Prolgolimab, Retifanlimab, Sasanlimab, Serplulimab, Sintilimab, Spartalizumab, Tislelizumab, Toripalimab, Ezabenlimab, Zimberelimab, Atezolizumab Lizumab), Avelumab, Cosibelimab, Sugemalimab, Durvalumab, Envafolimab, Suvratoxumab, Denosumab, Zilovertamab, Elotuzumab, Domvanalimab, Etigilimab, Ociperlima b) Tiragolumab, Vibostolimab, Surzebiclimab, Cobolimab, Sabatolimab, Concizumab, Marstacimab, Adalimumab, Golimumab, Infliximab, Certolizumab, Conatumumab, Gatuzumab, Tezepelumab, Gatipotuzumab, Cabiralizumab, Bevacizumab, Brolucizumab, Ranibizumab, Olinvacimab, Icrucumab, Ramucirumab, Caplacizumab, Abrilumab,Etrolizumab, Vedolizumab, Intetumumab, Natalizumab, Obrindatamab, Elranatamab, Linvoseltamab, Teclistamab Epcoritamab, Glofitamab, Mosunetuzumab, Odronextamab, Flotetuzumab, Vibecotamab, Catumaxomab, Cibisatamab, Talquetamab, Ubamatamab, Emfizatamab, Blinatumomab (inatumomab), amivantamab, emicizumab, xenocutuzumab, zanidatamab, tibulizumab, naptumomab, belantamab, pivekimab, praluzatamab, coltuximab, denintuzumab, loncustuzumab castuximab), ibritumomab, inotuzumab, epratuzumab, moxetumomab, brentuximab, gemtuzumab, vadastuximab, lorvotuzumab, polatuzumab, tusamitamab, telisotuzumab, donkey Rovalpituzumab, Depatuxizumab, Farletuzumab, Mirvetuximab, Disitamab, Anetumab, Enfortumab, Sacituzumab, Vobarilizumab, Cadonilimab, Vudalimab, Teboterimab,This may include, but is not limited to, ivonescimab, erfonrilimab, ozoralizumab, faricimab, vanucizumab, and navicixizumab.

[0031] The present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising a compound (chemical formula 1) or a carrier-drug conjugate. The pharmaceutical composition of the present invention can be administered to a target body requiring therapeutic action in a therapeutically effective amount.

[0032] In this invention, cancer refers to pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, epithelial ovarian cancer, ovarian germ cell tumor, male breast cancer, brain tumor, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, nasal cavity cancer / paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain tumor, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, Renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue tumors, malignant bone tumors, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary origin, gastric lymphoma, stomach cancer, gastric carcinoid tumor, gastrointestinal stromal tumor, Wilms' tumor, breast cancer, triple-negative breast cancer (TNBC), sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, intrauterine cancer This includes, but is not limited to, membrane cancer, uterine sarcoma, prostate cancer, metastatic bone tumors, metastatic brain tumors, mediastinal tumors, rectal cancer, rectal carcinoid tumors, vaginal cancer, spinal cord cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, hematological cancer, and thymic cancer. In this specification, the term "prevention" refers to any action by which the administration of the pharmaceutical composition suppresses the onset of cancer symptoms or delays their onset. The term "treatment" refers to any action by which the administration of the pharmaceutical composition improves or favorably alters cancer symptoms.

[0033] In this specification, “patient,” “subject,” and “subject” refer to animals such as mammals. In certain specific examples, the patient is a human. In other specific examples, the patient is a non-human animal such as a dog, cat, livestock (e.g., a horse, pig, or donkey), chimpanzee, or monkey.

[0034] As used in this invention, the term "therapeutably effective amount" refers to the amount of a compound or carrier-drug conjugate effective in treating or preventing cancer. Specifically, "therapeutably effective amount" means an amount sufficient to treat the disease with a reasonable benefit / risk balance applicable to medical treatment, and the effective dose level can be determined by factors including the individual's species and severity, age, sex, type of disease, drug activity, drug sensitivity, administration time, route of administration and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the medical field. The pharmaceutical compositions of this invention can be administered as individual therapeutic agents or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with commercially available therapeutic agents. They can also be administered as a single dose or multiple doses. It is important to administer the amount that provides the maximum effect with the minimum amount without side effects, taking all of the above factors into consideration. Since the compound represented by Chemical Formula 1 of the present invention, its derivatives, its isomers or pharmaceutically acceptable salts, or carrier drug complexes containing the same, exhibit dose-dependent effects, the dosage can be easily determined by those skilled in the art based on various factors such as the patient's condition, age, sex, and comorbidities. The active ingredient of the pharmaceutical composition of the present invention is highly safe and can be used at or above the determined dosage.

[0035] The synthesis of the compound of the present invention and its effects will be specifically confirmed below based on the examples. [Example 1] Synthesis of compounds with chemical formulas 2 and 3 1-1. Synthesis of A-fragments [ka]

[0036] In the above chemical structure, Boc2O represents di-tert-butyl dicarbonate.

[0037] Stage 1: [ka]

[0038] Methyl iodide (7.53 g, 3.30 mL, 2.4 eq, 53.1 mmol) was added to a solution of 2-(3-bromo-4-methoxyphenyl)acetonitrile (5.00 g, 1 eq, 22.1 mmol) in THF (dried) (30 mL). The reaction mixture was cooled to 4 °C, and sodium hydride (60% dispersion in mineral oil) (2.12 g, 60% Wt, 2.4 eq, 53.1 mmol) was added gradually. After the addition was complete, the reaction mixture was warmed to room temperature and stirred overnight. Methyl iodide (628 mg, 277 μL, 0.2 eq, 4.42 mmol) and sodium hydride (60% dispersion in mineral oil) (177 mg, 60% Wt, 0.2 eq, 4.42 mmol) were further added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was poured into ice water (~100 mL) and extracted with siRNA (3x). The combined organic layer was washed with brine, dried over Na₂SO₄, and concentrated under reduced pressure to obtain brown oil (6.47 g). This was purified by flash column chromatography (120 g SiO₂, 0-20% siRNA in heptane), and the product fraction was concentrated under reduced pressure to obtain colorless oil. Yield: 4.9g, 87%. 1H NMR (400MHz, CDCl3) δ7.62(d, J=2.6Hz, 1H), 7.40(dd, J=8.6, 2.5Hz, 1H), 6.90(d, J=8.7Hz, 1H), 3.89(s, 3H), 1.70(s, 6H). m / z 253.2 / 255.2[M+H] + Br isotope pattern.

[0039] Stage 2: [ka] A solution of 2-(3-bromo-4-methoxyphenyl)-2-methylpropanenitrile (4.68 g, 1 eq, 18.4 mmol) in dry DCM (40 mL) was cooled to -78°C. Diisobutylaluminum hydride (1 M in hexane) (3.40 g, 23.9 mL, 1.0 molar, 1.3 eq, 23.9 mmol) was added dropwise to the solution within 15 minutes. The reaction mixture was stirred at -78°C for 1.5 hours. Diisobutylaluminum hydride (1 M in hexane) (655 mg, 4.60 mL, 1.0 molar, 0.25 eq, 4.60 mmol) was added dropwise, and the reaction mixture was left to stir at -78°C for 30 minutes. The reaction mixture was warmed to 0°C, slowly quenched with 2 M HCl (60 mL), stirred at 0°C for 30 minutes, and then stirred at room temperature for 30 minutes. The mixture was diluted with water and extracted with DCM (3x). The combined organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain a solid (4.39 g). The crude product was suspended in DCM and filtered. The filtrate was purified by column chromatography (120 g SiO2, 0-20% siRNA in heptane). The product fraction was concentrated under reduced pressure to obtain a light-yellow oil (2.77 g, 58%). 1 H NMR (400MHz, CDCl3) δ9.44(s, 1H), 7.46(d, J=2.4Hz, 1H), 7.16(dd, J=8.6, 2.4Hz, 1H), 6.90(d, J=8.6Hz, 1H), 3.89(s, 3H), 1.44(s, 6H). m / z 257.2 / 259.2[M+H] + Br isotope pattern.

[0040] Stage 3: [ka] A 100 mL round-bottom flask was filled with potassium cyanide (0.56 g, 1 eq, 8.6 mmol) and water (13 mL). After cooling the solution to 0°C, methylamine hydrochloride (0.58 g, 1 eq, 8.6 mmol) was added. Then, a solution of 2-(3-bromo-4-methoxyphenyl)-2-methylpropanal (2.2 g, 1 eq, 8.6 mmol) in methanol (13 mL) was added. The resulting white suspension was stirred overnight at room temperature. The reaction mixture was diluted with H2O (40 mL) and extracted with CH2Cl2 (3 x 50 mL). The organic layers were combined, washed with brine (20 mL), and dried over Na2SO4. The solvent was removed under vacuum to obtain 3-(3-bromo-4-methoxyphenyl)-3-methyl-2-(methylamino)butanenitrile as a colorless oil. Yield: 2.56g, 89%. 1 H NMR (400MHz, CDCl3) δ7.57(d, J=2.4Hz, 1H), 7.36(dd, J=8.7, 2.4Hz, 1H), 6.89(d, J =8.7Hz, 1H), 3.89(s, 3H), 3.45-3.29(m, 1H), 2.48(s, 3H), 1.50(d, J=11.5Hz, 6H). m / z 297.0 / 299.0[M+H] + Br isotope pattern.

[0041] Stage 4: [ka] 3-(3-Bromo-4-methoxyphenyl)-3-methyl-2-(methylamino)butanenitrile (2.53 g, 1 eq, 8.51 mmol) in DMSO (45 mL) was added with lithium hydroxide monohydrate 1 M in water (1.71 g, 40.9 mL, 1.0 molar, 4.8 eq, 40.9 mmol) and hydrogen peroxide (4.63 g, 4.17 mL, 30% Wt, 4.8 eq, 40.9 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (385 mL) and extracted with ethyl acetate (2 x 385 mL). The organic layer was washed with water and brine. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to give 2.8 g of a colorless sticky solid. Purification by flash column chromatography (80 g SiO2, 0 - 5% methanol in dichloromethane) gave the product as a white foam. Yield: 1.35 g, 50%. 1 1H NMR (400 MHz, DMSO) δ 7.50 (d, J = 2.4 Hz, 1H), 7.35 - 7.23 (m, 2H), 7.05 - 6.97 (m, 2H), 3.81 (s, 3H), 2.99 - 2.90 (m, 1H), 2.07 (s, 3H), 1.44 (bs, 1H), 1.25 (d, J = 7.8 Hz, 6H). m / z 315.2 / 317.2 [M+H] + , Br isotope pattern.

[0042] Step 5:

Chemical Structure

[0043] Stage 6: [ka] A mixture of 3-(3-bromo-4-methoxyphenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate (350 mg, 1 eq, 841 μmol) and toluene (5.0 mL) was added to a 20 mL microwave vial. Ammonium hydroxide solution, 32% (9.21 g, 10.2 mL, 32% Wt, 100 eq, 84.1 mmol), was then added, followed by copper powder (160 mg, 3 eq, 2.52 mmol). The vial was sealed and stirred at 100°C for 6 hours. The reaction mixture was filtered, and the filtrate was evaporated. The residue was simultaneously evaporated with ethanol (3x) to remove water. The crude material was coated onto a hydromatrix and purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The product fraction was concentrated under reduced pressure to obtain 232 mg of green oil / foam. The product will be used directly in the next step. Yield: 67%. 1 H NMR (400MHz, CDCl3) δ7.23-5.96(m, 3H), 5.20-4.70(m, 1H), 4.05-3.55(m, 3H), 3.01-2.41(m, 4H), 1.93-1.09(m, 17H). m / z 705.2[2M+H] +

[0044] Stage 7: [ka] To a solution of 3-(3-amino-4-methoxyphenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate (205 mg, 1 eq, 582 μmol) in tetrahydrofuran (1.5 mL), Alloc-OSu (232 mg, 2 eq, 1.16 mmol) was added, followed by the addition of triethylamine (177 mg, 243 μL, 3 eq, 1.75 mmol). The reaction mixture was stirred overnight at room temperature. The solvent was removed under reduced pressure, and the residue was purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The product fraction was concentrated to obtain 143 mg (42%) of a light brown oil. NMR showed rotomerity. 1 H NMR (400MHz, CDCl3) δ8.20(s, 1H), 7.24(s, 1H), 7.11-6.97(m, 1H), 6.79(d, J=8.6Hz, 1H), 6.05-5.92(m, 1H), 5.41-5.33 (m, 1H), 5.28-5.23(m, 1H), 4.94(s, 1H), 4.70-4.64(m, 2H), 3.84(s, 3H), 2.77(s, 1H), 2.65(s, 2H), 1.56-1.40(m, 16H). m / z 459.2[M+Na] +

[0045] 1-2. Synthesis of BCD fragments [ka]

[0046] Stage 1: [ka] HBTU (9.0 g, 1.1 eq, 24 mmol) was gradually added to a stirred solution of N-(tert-butoxycarbonyl)-N-methyl-L-valine (5.0 g, 1 eq, 22 mmol), N,O-dimethylhydroxylamine hydrochloride (2.3 g, 1.1 eq, 24 mmol), and DIPEA (5.6 g, 7.5 mL, 2 eq, 43 mmol) in dichloromethane (50 mL). The reaction mixture was stirred overnight at room temperature. The reaction mixture was washed with water. The organic layer was separated and dried over Na₂SO₄. The organic layer was then evaporated and purified by column chromatography (120 g SiO₂, heptane with 0-40% ethyl acetate). The product was obtained as a clear, colorless oil. Yield: 3.15g, 53%. 1 H NMR (400MHz, DMSO-d6) rotamer mixture δ4.94-4.44(m, 1H), 3.64(d, J=10.0Hz, 3H), 3.19-3.0 1(m, 3H), 2.74-2.60(m, 3H), 2.21-2.04(m, 1H), 1.48-1.33(m, 9H), 0.89-0.73(m, 6H).

[0047] Stage 2: [ka] At -78°C, 2.4 M lithium aluminum hydride in THF (69.2 mg, 759 μL, 2.4 molar, 1 eq, 1.82 mmol) was added in part to a solution of tert-butyl(S)-(1-(methoxy(methyl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (500 mg, 1 eq, 1.82 mmol) in THF (2.5 mL). The reaction mixture was then heated to 0°C and stirred for 15 minutes. The reaction mixture was poured into an ice-cold 0.25 M aqueous solution of potassium bisulfate (496 mg, 14.6 mL, 0.25 molar, 2 eq, 3.64 mmol) while stirring, and the layers were separated. The aqueous layer was extracted with diethyl ether (3x). The combined organic layers were washed with 1M HCl (2x), saturated aqueous sodium bicarbonate (2x), and brine, dried over sodium sulfate, and then concentrated (bath temperature below 20°C). The crude aldehyde was used directly in the next step without purification.

[0048] Stage 3: [ka] A solution of tert-butyl(S)-methyl(3-methyl-1-oxobutan-2-yl)carbamate (392 mg, 1 eq, 1.82 mmol) in dichloromethane (2.5 mL) was prepared by adding ethyl 2-(triphenyl-λ) 5 Phosphanylidene propanoate (857 mg, 1.3 eq, 2.37 mmol) was added, and the reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with water and extracted with diethyl ether (3x). The combined organic layer was washed with brine, dried over sodium sulfate, and then concentrated to obtain 1.0 g of yellow oil. It was purified by flash column chromatography (40 g SiO2, 0-10% ethyl acetate in heptane) to obtain 237 mg of colorless oil. 1H NMR (400MHz, CDCl3) δ6.70-6.60(m, 1H), 4.64-4.25(m, 1H), 4.21(q, J=7.1Hz, 2H), 2.80 -2.62(m, 3H), 1.96-1.81(m, 4H), 1.45(s, 9H), 1.31(t, J=7.0Hz, 3H), 0.96-0.79(m, 6H). m / z 322.2[M+Na] +

[0049] Stage 4: [ka] Trifluoroacetic acid (2.5 mL) was added to a solution of ethyl(S,E)-4-((tert-butoxycarbonyl)(methyl)amino)-2,5-dimethylhexa-2-enoate (235 mg, 1 eq, 785 μmol) in dichloromethane (2.5 mL). The reaction mixture was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure, and the residue was simultaneously evaporated with dichloromethane (6x) to obtain 325 mg of a slightly yellow, turbid oil. The product was used directly in the next reaction. 1 H NMR (400MHz, CDCl3) δ8.86(d, J=174.6Hz, 1H), 6.63-6.56(m, 1H), 4.33-4.17(m, 2H, coincides with H2O peak), 3.82-3.69(m, 1H), 2.62(s, 3H), 2.25-2.13(m, 1H), 1.95(s, 3H), 1.33(t, J=7.1Hz, 3H), 1.02(dd, J=12.2, 6.8Hz, 6H). m / z 200.0[M+H] +

[0050] Stage 5: [ka] DIPEA (101 mg, 137 μL, 1 eq, 785 μmol) was added to a solution of N-Boc-L-tert-leucine (272 mg, 1.5 eq, 1.18 mmol) and HATU (448 mg, 1.5 eq, 1.18 mmol) in N,N-dimethylformamide (3.0 mL). After stirring the mixture for 5 minutes, a solution of ethyl(S,E)-2,5-dimethyl-4-(methylamino)hexa-2-enoate 2,2,2-trifluoroacetate (246 mg, 1 eq, 785 μmol) and DIPEA (507 mg, 684 μL, 5 eq, 3.93 mmol) in peptide grade N,N-dimethylformamide (1.5 mL) was added dropwise. The mixture was stirred at room temperature for 90 minutes. The reaction mixture was diluted with ethyl acetate, washed with water (2x) and brine (2x), dried over sodium sulfate, and concentrated to obtain 478 mg of yellow oil. After purification by flash column chromatography (24 g SiO2, 0-50% ethyl acetate in heptane), the product fraction was concentrated to obtain 272 mg (76%) of colorless oil. 1 H NMR (400MHz, CDCl3) δ6.68-6.60(m, 1H), 5.22(d, J=10.1Hz, 1H), 5.15-5.06(m, 1H), 4.43(d, J=10.1Hz, 1H), 4.20(q, J =7.1Hz, 2H), 2.99(s, 3H), 1.90(m, 4H), 1.42(s, 9H), 1.31(t, J=7.1Hz, 3H), 0.96(s, 9H), 0.86(dd, J=18.3, 6.6Hz, 6H). m / z 413.2[M+H] +

[0051] Stage 6: [ka] To a solution of ethyl(S,E)-4-((S)-2-((tert-butoxycarbonyl)amino)-N,3,3-trimethylbutanamide)-2,5-dimethylhexa-2-enoate (260 mg, 90.7% Wt, 1 eq, 572 μmol) in dichloromethane (4.0 mL), trifluoroacetic acid (4.0 mL) was added, and the reaction mixture was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure, and the residue was simultaneously evaporated with dichloromethane (6x) to obtain 329 mg of colorless oil. 1 H NMR (400MHz, CDCl3) δ7.92(bs, 2H), 6.69-6.62(m, 1H), 5.70(bs, 3H), 5.00(t, J=10.1Hz, 1H), 4.27(s, 1H), 4.22(q, J=7. 1Hz, 2H), 2.98(s, 3H), 2.03-1.93(m, 1H), 1.91(s, 3H), 1.31(t, J=7.1Hz, 3H), 1.08(s, 9H), 0.87(dd, J=11.3, 6.5Hz, 6H). m / z 313.4[M+H] +

[0052] 1-3. Synthesis of Compounds of Chemical Formulas 2 and 3 [ka]

[0053] Stage 1: [ka] To a solution of 3-(3-(((allyloxy)carbonyl)amino)-4-methoxyphenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate [Fragment A] (128 mg, 85% Wt, 1 eq, 249 μmol) in N,N-dimethylformamide (1.8 mL), ethyl (S,E)-4-((S)-2-amino-N,3,3-trimethylbutanamide)-2,5-dimethylhexa-2-enoate 2,2,2-trifluoroacetate (142 mg, 75% Wt, 1 eq, 249 μmol) was added, followed by the addition of PyBOP (143 mg, 1.1 eq, 274 μmol). The reaction mixture was cooled to 0°C, and DIPEA (129 mg, 174 μL, 4 eq, 997 μmol) was added. The resulting clear solution was then stirred overnight at room temperature. The reaction mixture was used directly for purification using acidic preparative MPLC (Luna 40-80). After combining the product fractions, they were freeze-dried and then evaporated with DCM. An orange oil was obtained. Yield: 98 mg, 54%. The product was obtained as a mixture of diastereoisomers. 1 H NMR (400MHz, CDCl3) δ8.22(s, 1H), 7.25-7.17(m, 1H), 7.15-6.99(m, 1H), 6.78(d, J=8.6Hz) , 1H), 6.65-6.57(m, 1H), 6.38-6.06(m, 1H), 6.06-5.91(m, 1H), 5.41-5.33(m, 1H), 5.29-5 .23(m, 1H), 5.12-4.81(m, 2H), 4.75-4.59(m, 3H), 4.23-4.15(m, 2H), 3.82(s, 3H), 2.97-2 .79(m, 6H), 1.91-1.80(m, 4H), 1.54-1.37(m, 14H), 1.33-1.22(m, 4H), 0.90-0.72(m, 16H). m / z 731.6[M+H] +

[0054] Stage 2: [ka] A solution of ethyl(9S,12S,E)-6-(2-(3-(((allyloxy)carbonyl)amino)-4-methoxyphenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-oate (90.5 mg, 1 eq, 124 μmol) was bubbled with nitrogen in tetrahydrofuran (6.0 mL). Pd(PPh3)4 (7.1 mg, 0.05 eq, 6.19 μmol) was added, followed by the addition of tri-n-butyltin hydride (72.1 mg, 66.1 μL, 2 eq, 248 μmol). The mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The solvent was removed under reduced pressure, and the crude substance was purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The fractions were combined and concentrated to obtain 79 mg of orange oil. This was purified by flash column chromatography (4 g SiO2, 0-40% ethyl acetate in heptane) to obtain the product as a white solid. Yield: 56 mg, 70%. 1 H NMR (400MHz, CDCl3) δ7.01-6.55(m, 4H), 6.28-5.98(m, 1H), 5.11-5.00(m, 1H), 4.60(dd, J=39.1, 9.2Hz, 1H), 4.24- 4.14(m, 2H), 3.86-3.71(m, 5H), 2.98-2.81(m, 6H), 1.86(d, J=10.1Hz, 4H), 1.51-1.18(m, 19H), 0.91-0.68(m, 16H). m / z 647.2[M+H] +

[0055] Stage 3: [ka] A solution of lithium hydroxide monohydrate (16.4 mg, 5 eq, 390 μmol) in water (1.0 mL) was added to a solution of ethyl (9S,12S,E)-6-(2-(3-amino-4-methoxyphenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-oate (50.5 mg, 1 eq, 78.1 μmol) in methanol (2.0 mL). The solution turned yellow. The reaction mixture was stirred overnight at room temperature. After neutralizing the reaction mixture with acetic acid, it was extracted with ethyl acetate (3x). The organic layer was washed with brine, dried over sodium sulfate, concentrated, and co-evaporated with dichloromethane (2x) to obtain a beige solid. Yield: 47 mg, 97% m / z 619.2[M+H] +

[0056] Stage 4: [ka] To a solution of (9S,12S,E)-6-(2-(3-amino-4-methoxyphenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-acid (46 mg, 1 eq, 74 μmol) in dichloromethane (1.0 mL), 10% TFA in dichloromethane (1.0 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. The mixture was cooled to 0°C, and TFA (740 mg, 500 μL, 87 eq, 6.49 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, and the residue was purified using acidic preparative MPLC (Luna 5-40). The product fractions were combined and then freeze-dried. Yield: 19 mg, 50%. The product was obtained as a mixture of diastereoisomers. The diastereoisomers were separated by RP-HPLC to produce S,S,S (5 mg) and R,S,S (7 mg) isomers. 1 H NMR (400MHz, DMSO-d6) δ7.76 (s, 1H), 6.76-6.54 (m, 4H), 5.31-4.01 (m, 4H), 3.73 (d, J=2.9Hz, 3H), 3.30 (s, 2H), 3.16- 2.89 (m, 4H), 2.12-1.89 (m, 4H), 1.78 (d, J=1.4Hz, 3H), 1.28-1.10 (m, 6H), 0.90 (d, J=24.6Hz, 9H), 0.83-0.67 (m, 6H). m / z 519.4[M+H] +

[0057] [Example 2] Synthesis of compounds with chemical formulas 4 and 5

change

change

[0058] Stage 1:

change

[0059] Stage 2: [ka] A solution of 2-(3-bromo-4-methylphenyl)-2-methylpropanenitrile (4.38 g, 1 eq, 18.4 mmol) in dry DCM (40 mL) was cooled to -78°C. Diisobutylaluminum hydride (1 M in hexane) (3.40 g, 23.9 mL, 1.0 molar, 1.3 eq, 23.9 mmol) was added dropwise to the solution within 15 minutes. The reaction mixture was stirred at -78°C for 1.5 hours. Diisobutylaluminum hydride (1 M in hexane) (655 mg, 4.60 mL, 1.0 molar, 0.25 eq, 4.60 mmol) was added dropwise, and the reaction mixture was left to stir at -78°C for 30 minutes. The reaction mixture was warmed to 0°C, slowly quenched with 2 M HCl (60 mL), stirred at 0°C for 30 minutes, and then stirred at room temperature for 30 minutes. The mixture was diluted with water and extracted with DCM (3x). The combined organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain a solid (4.39 g). The crude product was suspended in DCM and filtered. The filtrate was purified by column chromatography (120 g SiO2, 0-20% siRNA in heptane). The product fraction was concentrated under reduced pressure to obtain a light-yellow oil (2.68 g, 60%). 1 H NMR (400MHz, CDCl3) δ9.42(s, 1H), 7.41(d, J=2.4Hz, 1H), 7.13(dd, J=8.6, 2.4Hz, 1H), 6.85(d, J=8.6Hz, 1H), 2.33(s, 3H), 1.43(s, 6H). m / z 241.2 / 243.2[M+H] + Br isotope pattern.

[0060] Stage 3: [ka] Potassium cyanide (0.56 g, 1 eq, 8.6 mmol) and water (13 mL) were packed into a 100 mL round-bottom flask. After cooling the solution to 0°C, methylamine hydrochloride (0.58 g, 1 eq, 8.6 mmol) was added. Then, a solution of 2-(3-bromo-4-methylphenyl)-2-methylpropanal (2.25 g, 1 eq, 8.6 mmol) in methanol (13 mL) was added. The resulting white suspension was stirred overnight at room temperature. The reaction mixture was diluted with H2O (40 mL) and extracted with CH2Cl2 (3 x 50 mL). The organic layers were combined, washed with brine (20 mL), and dried over Na2SO4. The solvent was removed under vacuum to obtain 3-(3-bromo-4-chlorophenyl)-3-methyl-2-(methylamino)butanenitrile as a colorless oil. Yield: 2.2g, 86%. 1 H NMR (400MHz, CDCl3) δ7.52(d, J=2.4Hz, 1H), 7.31(dd, J=8.7, 2.4Hz, 1H), 6.84(d, J =8.7Hz, 1H), 3.43-3.31(m, 1H), 2.46(s, 3H), 2.33(s, 3H), 1.50(d, J=11.5Hz, 6H). m / z 281.0 / 283.0[M+H] + Br isotope pattern.

[0061] Stage 4: [ka] 3-(3-bromo-4-methylphenyl)-3-methyl-2-(methylamino)butanenitrile (2.39 g, 1 eq, 8.51 mmol) in DMSO (45 mL) was mixed with 1 M lithium hydroxide monohydrate (1.71 g, 40.9 mL, 1.0 molar, 4.8 eq, 40.9 mmol) and hydrogen peroxide (4.63 g, 4.17 mL, 30% Wt, 4.8 eq, 40.9 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (385 mL) and extracted with ethyl acetate (2 x 385 mL). The organic layer was washed with water and brine. The organic layer was dried over sodium sulfate and then concentrated under reduced pressure to obtain 2.5 g of a colorless, viscous solid. The product was purified by flash column chromatography (80 g SiO2, 0-5% methanol in dichloromethane) to obtain the product as a white foam. Yield: 1.23g, 48%. 1 H NMR (400MHz, DMSO) δ7.46(d, J=2.4Hz, 1H), 7.33-7.21(m, 2H), 7.02-6.95(m, 2H), 2.97-2.87(m, 1H), 2.30(s, 3H), 2.07(s, 3H), 1.44(bs, 1H), 1.23(d, J=7.8Hz, 6H). m / z 299.2 / 301.2[M+H] + Br isotope pattern.

[0062] Stage 5: [ka] 3-(3-bromo-4-methylphenyl)-3-methyl-2-(methylamino)butanamide (1.24 g, 1 eq, 4.16 mmol) was mixed with dichloromethane (15 mL). Next, Boc2O (3.17 g, 3.5 eq, 14.5 mmol) was added. After a few minutes, a clear, colorless solution was obtained. The reaction mixture was stirred at room temperature for 3 days. DIPEA (591 mg, 787 μL, 1.1 eq, 4.57 mmol) and DMAP (50.8 mg, 0.1 eq, 416 μmol) were added. The reaction mixture was stirred at room temperature for 2 hours. Water (45 mL) was added and extracted with heptane (3 x 50 mL). The organic layer was washed with water (25 mL) and brine (25 mL). The organic layer was dried over sodium sulfate and concentrated dry. 2.8 g of yellow oil was obtained and dissolved in THF (6.5 mL). 5M sodium hydroxide (831 mg, 4.16 mL, 5.0 molar, 5 eq, 20.8 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. Then, water (25 mL) was added. The mixture was filtered, and the residue was washed with 0.5 M NaOH (25 mL). THF was removed under reduced pressure. The aqueous solution was washed with heptane, neutralized with solid citric acid, and extracted with ethyl acetate (3x). The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to obtain 1.35 g of colorless oil. After purification by column chromatography (40 g SiO2, 0-50% ethyl acetate in heptane), the product fraction was concentrated under reduced pressure to obtain a white foam. Yield: 1.04g, 62%. 1 H NMR (400MHz, CDCl3) δ7.55-7.46(m, 1H), 7.33-7.22(m, 1H), 6.78(d, J=8.7Hz , 1H), 5.03-4.80(m, 1H), 2.82-2.60(m, 3H), 2.33(s, 3H), 1.53-1.35(m, 15H). m / z 398.2 / 400.2[MH] - Br isotope pattern.

[0063] Stage 6: [ka] A mixture of 3-(3-bromo-4-methylphenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate (354 mg, 1 eq, 841 μmol) and toluene (5.0 mL) was added to a 20 mL microwave vial. Ammonium hydroxide solution, 32% (9.21 g, 10.2 mL, 32% Wt, 100 eq, 84.1 mmol), was then added, followed by copper powder (160 mg, 3 eq, 2.52 mmol). The vial was sealed and stirred at 100°C for 6 hours. The reaction mixture was filtered, and the filtrate was evaporated. The residue was simultaneously evaporated with ethanol (3x) to remove water. The crude material was coated onto a hydromatrix and purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The product fraction was concentrated under reduced pressure to obtain 232 mg of green oil / foam. The product will be used directly in the next step. Yield: 70%. 1 H NMR (400MHz, CDCl3) δ7.18-5.90(m, 3H), 5.15-4.65(m, 1H), 2.89-2.39(m, 4H), 2.33-2.28(m, 3H) 1.90-1.05(m, 17H). m / z 673.2[2M+H] +

[0064] Stage 7: [ka] To a solution of 3-(3-amino-4-methylphenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate (196 mg, 1 eq, 582 μmol) in tetrahydrofuran (1.5 mL), Alloc-OSu (232 mg, 2 eq, 1.16 mmol) was added, followed by the addition of triethylamine (177 mg, 243 μL, 3 eq, 1.75 mmol). The reaction mixture was stirred overnight at room temperature. The solvent was removed under reduced pressure, and the residue was purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The product fraction was concentrated to obtain 110 mg (45%) of a light brown oil. NMR showed rotomerity. 1 H NMR (400MHz, CDCl3) δ8.18(s, 1H), 7.21(s, 1H), 7.08-6.95(m, 1H), 6.75(d, J=8.6Hz, 1H), 6.02-5.90(m, 1H), 5.39-5.31 (m, 1H), 5.25-5.20(m, 1H), 4.91(s, 1H), 4.68-4.62(m, 2H), 2.75(s, 1H), 2.63(s, 2H), 2.31(s, 3H), 1.52-1.38(m, 16H). m / z 443.2[M+Na] +

[0065] 2-2. Synthesis of Compounds of Chemical Formulas 4 and 5 [ka]

[0066] Stage 1: [ka] To a solution of 3-(3-(((allyloxy)carbonyl)amino)-4-methylphenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate [fragment A] (123 mg, 85% Wt, 1 eq, 249 μmol) in N,N-dimethylformamide (1.8 mL), ethyl (S,E)-4-((S)-2-amino-N,3,3-trimethylbutanamide)-2,5-dimethylhexa-2-enoate 2,2,2-trifluoroacetate [fragment BCD] (142 mg, 75% Wt, 1 eq, 249 μmol) was added, followed by the addition of PyBOP (143 mg, 1.1 eq, 274 μmol). The reaction mixture was cooled to 0°C, and DIPEA (129 mg, 174 μL, 4 eq, 997 μmol) was added. The resulting clear solution was stirred overnight at room temperature. The reaction mixture was used directly for purification using acidic preparative MPLC (Luna 40-80). After combining the product fractions, they were freeze-dried and then evaporated with DCM. An orange oil was obtained. Yield: 108 mg, 60%. The product was obtained as a mixture of diastereoisomers. 1H NMR (400MHz, CDCl3) δ8.20(s, 1H), 7.23-7.12(m, 1H), 7.12-6.96(m, 1H), 6.75(d, J=8.6Hz) , 1H), 6.63-6.54(m, 1H), 6.35-6.03(m, 1H), 6.03-5.88(m, 1H), 5.38-5.30(m, 1H), 5.25-5 .20(m, 1H), 5.10-4.78(m, 2H), 4.72-4.55(m, 3H), 4.20-4.12(m, 2H), 2.95-2.75(m, 6H), 2 .31(s, 3H), 1.89-1.78(m, 4H), 1.52-1.32(m, 14H), 1.31-1.20(m, 4H), 0.89-0.70(m, 16H). m / z 715.6[M+H] +

[0067] Stage 2: [ka] A solution of ethyl(9S,12S,E)-6-(2-(3-(((allyloxy)carbonyl)amino)-4-methylphenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-oate (88.6 mg, 1 eq, 124 μmol) was bubbled with nitrogen in tetrahydrofuran (6.0 mL). Pd(PPh3)4 (7.1 mg, 0.05 eq, 6.19 μmol) was added, followed by the addition of tri-n-butyltin hydride (72.1 mg, 66.1 μL, 2 eq, 248 μmol). The mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The solvent was removed under reduced pressure, and the crude substance was purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The fractions were combined and concentrated to obtain 79 mg of orange oil. This was purified by flash column chromatography (4 g SiO2, 0-40% ethyl acetate in heptane) to obtain the product as a white solid. Yield: 53 mg, 68%. 1H NMR (400MHz, CDCl3) δ6.98-6.52(m, 4H), 6.25-5.95(m, 1H), 5.06-4.95(m, 1H), 4.55(dd, J=39.1, 9.2Hz, 1H), 4.20-4.10(m , 2H), 3.83-3.68(m, 2H), 2.95-2.78(m, 6H), 2.22(s, 3H), 1.86(d, J=10.1Hz, 4H), 1.51-1.18(m, 19H), 0.91-0.68(m, 16H). m / z 631.2[M+H] +

[0068] Stage 3: [ka] A solution of lithium hydroxide monohydrate (16.4 mg, 5 eq, 390 μmol) in water (1.0 mL) was added to a solution of ethyl (9S,12S,E)-6-(2-(3-amino-4-methylphenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-oate (49 mg, 1 eq, 78.1 μmol) in methanol (2.0 mL). The solution turned yellow. The reaction mixture was stirred overnight at room temperature. After neutralizing the reaction mixture with acetic acid, it was extracted with ethyl acetate (3x). The organic layer was washed with brine, dried over sodium sulfate, concentrated, and co-evaporated with dichloromethane (2x) to obtain a beige solid. Yield: 46 mg, 98% m / z 603.2[M+H] +

[0069] Stage 4: [ka] To a solution of (9S,12S,E)-6-(2-(3-amino-4-methylphenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-acid (46 mg, 1 eq, 74 μmol) in dichloromethane (1.0 mL), 10% TFA in dichloromethane (1.0 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. The mixture was cooled to 0°C, and TFA (740 mg, 500 μL, 87 eq, 6.49 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, and the residue was purified using acidic preparative MPLC (Luna 5-40). The product fractions were combined and then freeze-dried. Yield: 19 mg, 50%. The product was obtained as a mixture of diastereoisomers. The diastereoisomers were separated by RP-HPLC to produce S,S,S (5 mg) and R,S,S (7 mg) isomers. 1 H NMR (400MHz, DMSO-d6) δ7.72(s, 1H), 6.73-6.51(m, 4H), 5.30-4.00(m, 4H), 3.28(s, 2H), 3.14-2.86(m, 4H), 2.18(d, J =2.9Hz, 3H), 2.10-1.84(m, 4H), 1.74(d, J=1.4Hz, 3H), 1.24-1.08(m, 6H), 0.89(d, J=24.6Hz, 9H), 0.82-0.65(m, 6H). m / z 503.4[M+H] +

[0070] [Example 3] Synthesis of compounds with chemical formulas 6 and 7 [ka]

[0071] 3-1. Synthesis of Fragment A [ka]

[0072] Stage 1: [ka] Methyl iodide (7.53 g, 3.30 mL, 2.4 eq, 53.1 mmol) was added to a solution of 2-(3-bromo-4-chlorophenyl)acetonitrile (5.00 g, 1 eq, 22.1 mmol) in THF (dried) (30 mL). The reaction mixture was cooled to 4 °C, and sodium hydride (60% dispersion in mineral oil) (2.12 g, 60% Wt, 2.4 eq, 53.1 mmol) was added little by little. After the addition was complete, the reaction mixture was warmed to room temperature and stirred overnight. Methyl iodide (628 mg, 277 μL, 0.2 eq, 4.42 mmol) and sodium hydride (60% dispersion in mineral oil) (177 mg, 60% Wt, 0.2 eq, 4.42 mmol) were further added, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was poured into ice water (~100 mL) and extracted with siRNA (3x). The combined organic layer was washed with brine, dried over Na₂SO₄, and concentrated under reduced pressure to obtain brown oil (6.47 g). This was purified by flash column chromatography (120 g SiO₂, 0-20% siRNA in heptane), and the product fraction was concentrated under reduced pressure to obtain colorless oil. Yield: 4.7g, 82%. 1 H NMR (400MHz, CDCl3) δ7.52 (d, J=2.6Hz, 1H), 7.34 (dd, J=8.6, 2.5Hz, 1H), 7.15 (d, J=8.7Hz, 1H), 1.70 (s, 6H). m / z 257.2 / 259.2[M+H] + Br isotope pattern.

[0073] Stage 2: [ka] A solution of 2-(3-bromo-4-chlorophenyl)-2-methylpropanenitrile (4.76 g, 1 eq, 18.4 mmol) in dry DCM (40 mL) was cooled to -78°C. Diisobutylaluminum hydride (1 M in hexane) (3.40 g, 23.9 mL, 1.0 molar, 1.3 eq, 23.9 mmol) was added dropwise to the solution within 15 minutes. The reaction mixture was stirred at -78°C for 1.5 hours. Diisobutylaluminum hydride (1 M in hexane) (655 mg, 4.60 mL, 1.0 molar, 0.25 eq, 4.60 mmol) was added dropwise, and the reaction mixture was left to stir at -78°C for 30 minutes. The reaction mixture was warmed to 0°C, slowly quenched with 2 M HCl (60 mL), stirred at 0°C for 30 minutes, and then stirred at room temperature for 30 minutes. The mixture was diluted with water and extracted with DCM (3x). The combined organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain a solid (4.39 g). The crude product was suspended in DCM and filtered. The filtrate was purified by column chromatography (120 g SiO2, 0-20% siRNA in heptane). The product fraction was concentrated under reduced pressure to obtain a light-yellow oil (2.49 g, 52%). 1 H NMR (400MHz, CDCl3) δ9.45(s, 1H), 7.51(d, J=2.3Hz, 1H), 7.35(dd, J=8.5, 2.3Hz, 1H), 7.18(d, J=8.5Hz, 1H), 1.45(s, 6H). m / z 260.2 / 262.2[M+H] + Br isotope pattern.

[0074] Stage 3: [ka] A 100 mL round-bottom flask was packed with potassium cyanide (0.56 g, 1 eq, 8.6 mmol) and water (13 mL). After cooling the solution to 0°C, methylamine hydrochloride (0.58 g, 1 eq, 8.6 mmol) was added. Then, a solution of 2-(3-bromo-4-chlorophenyl)-2-methylpropanal (2.25 g, 1 eq, 8.6 mmol) in methanol (13 mL) was added. The resulting white suspension was stirred overnight at room temperature. The reaction mixture was diluted with H2O (40 mL) and extracted with CH2Cl2 (3 x 50 mL). The organic layers were combined, washed with brine (20 mL), and dried over Na2SO4. The solvent was removed under vacuum to obtain 3-(3-bromo-4-chlorophenyl)-3-methyl-2-(methylamino)butanenitrile as a colorless oil. Yield: 2.08g, 80%. 1 H NMR (400MHz, CDCl3) δ7.58(d, J=2.4Hz, 1H), 7.33(dd, J=8.7, 2.4Hz, 1H), 7.09(d, J=8.7Hz, 1H), 3.45-3.35(m, 1H), 2.48(s, 3H), 1.52(d, J=11.5Hz, 6H). m / z 300.4 / 302.4[M+H] + Br isotope pattern.

[0075] Stage 4: [ka] 3-(3-bromo-4-chlorophenyl)-3-methyl-2-(methylamino)butanenitrile (2.57 g, 1 eq, 8.51 mmol) in DMSO (45 mL) was mixed with 1 M lithium hydroxide monohydrate (1.71 g, 40.9 mL, 1.0 molar, 4.8 eq, 40.9 mmol) and hydrogen peroxide (4.63 g, 4.17 mL, 30% Wt, 4.8 eq, 40.9 mmol) in water. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (385 mL) and extracted with ethyl acetate (2 x 385 mL). The organic layer was washed with water and brine. The organic layer was dried over sodium sulfate and then concentrated under reduced pressure to obtain 2.5 g of a colorless, viscous solid. The product was purified by flash column chromatography (80 g SiO2, 0-5% methanol in dichloromethane) to obtain the product as a white foam. Yield: 1.23g, 45%. 1 H NMR (400MHz, DMSO) δ7.52(d, J=2.4Hz, 1H), 7.38-7.28(m, 2H), 7.18-7.13(m, 2H), 3.00-2.95(m, 1H), 2.10(s, 3H), 1.48(bs, 1H), 1.25(d, J=7.8Hz, 6H). m / z 318.4 / 320.4[M+H] + Br isotope pattern.

[0076] Stage 5: [ka] 3-(3-bromo-4-chlorophenyl)-3-methyl-2-(methylamino)butanamide (1.33 g, 1 eq, 4.16 mmol) was mixed with dichloromethane (15 mL). Next, Boc2O (3.17 g, 3.5 eq, 14.5 mmol) was added. After a few minutes, a clear, colorless solution was obtained. The reaction mixture was stirred at room temperature for 3 days. DIPEA (591 mg, 787 μL, 1.1 eq, 4.57 mmol) and DMAP (50.8 mg, 0.1 eq, 416 μmol) were added. The reaction mixture was stirred at room temperature for 2 hours. Water (45 mL) was added and extracted with heptane (3 x 50 mL). The organic layer was washed with water (25 mL) and brine (25 mL). The organic layer was dried over sodium sulfate and concentrated dry. 2.8 g of yellow oil was obtained and dissolved in THF (6.5 mL). The reaction mixture was stirred at room temperature for 2 hours after adding 5M sodium hydroxide (831 mg, 4.16 mL, 5.0 molar, 5 eq, 20.8 mmol). Then, water (25 mL) was added. The mixture was filtered, and the residue was washed with 0.5 M NaOH (25 mL). THF was removed under reduced pressure. The aqueous solution was washed with heptane, neutralized with solid citric acid, and extracted with ethyl acetate (3x). The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to obtain 1.37 g of colorless oil. After purification by column chromatography (40 g SiO2, 0-50% ethyl acetate in heptane), the product fraction was concentrated under reduced pressure to obtain a white foam. Yield: 1.13g, 65%. 1 ¹H NMR (400 MHz, CDCl3) δ 7.58-7.48 (m, 1H), 7.39-7.32 (m, 1H), 7.18 (d, J=8.7 Hz, 1H), 5.13-4.90 (m, 1H), 2.92-2.70 (m, 3H), 1.58-1.45 (m, 15H). m / z 419.2 / 421.2 [MH]-, Br isotope pattern.

[0077] Stage 6: [ka] A mixture of 3-(3-bromo-4-chlorophenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate (354 mg, 1 eq, 841 μmol) and toluene (5.0 mL) was added to a 20 mL microwave vial. Ammonium hydroxide solution, 32% (9.21 g, 10.2 mL, 32% Wt, 100 eq, 84.1 mmol), was then added, followed by copper powder (160 mg, 3 eq, 2.52 mmol). The vial was sealed and stirred at 100°C for 6 hours. The reaction mixture was filtered, and the filtrate was evaporated. The residue was simultaneously evaporated with ethanol (3x) to remove water. The crude material was coated onto a hydromatrix and purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The product fraction was concentrated under reduced pressure to obtain 232 mg of green oil / foam. The product will be used directly in the next step. Yield: 60%. 1 H NMR (400MHz, CDCl3) δ7.28-6.90(m, 2H), 6.85-6.30(m, 1H), 5.25-4.85(m, 1H), 2.95-2.59(m, 4H), 1.92-1.10(m, 17H). m / z 713.4[2M+H] +

[0078] Stage 7: [ka] Alloc-OSu (232 mg, 2 eq, 1.16 mmol) was added to a solution of 3-(3-amino-4-chlorophenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate (205 mg, 1 eq, 582 μmol) in tetrahydrofuran (1.5 mL), followed by the addition of triethylamine (177 mg, 243 μL, 3 eq, 1.75 mmol). The reaction mixture was stirred overnight at room temperature. The solvent was removed under reduced pressure, and the residue was purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The product fraction was concentrated to obtain 120 mg (47%) of a light brown oil. NMR showed rotomerity. 1 H NMR (400MHz, CDCl3) δ8.28(s, 1H), 7.31(s, 1H), 7.25-7.10(m, 1H), 6.95(d, J=8.6Hz, 1H), 6.12-6.05(m, 1H), 5.4 5-5.36(m, 1H), 5.28-5.23(m, 1H), 4.96(s, 1H), 4.72-4.68(m, 2H), 2.85(s, 1H), 2.73(s, 2H), 1.55-1.42(m, 16H). m / z 463.2[M+Na] +

[0079] 3-2. Synthesis of Compounds of Chemical Formulas 6 and 7 [ka]

[0080] Stage 1: [ka] To a solution of 3-(3-(((allyloxy)carbonyl)amino)-4-chlorophenyl)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutyrate [fragment A] (125 mg, 88% Wt, 1 eq, 249 μmol) in N,N-dimethylformamide (1.8 mL), ethyl (S,E)-4-((S)-2-amino-N,3,3-trimethylbutanamide)-2,5-dimethylhexa-2-enoate 2,2,2-trifluoroacetate (142 mg, 75% Wt, 1 eq, 249 μmol) [fragment BCD] was added, followed by the addition of PyBOP (143 mg, 1.1 eq, 274 μmol). The reaction mixture was cooled to 0°C, and DIPEA (129 mg, 174 μL, 4 eq, 997 μmol) was added. The resulting clear solution was stirred overnight at room temperature. The reaction mixture was used directly for purification using acidic preparative MPLC (Luna 40-80). After combining the product fractions, they were freeze-dried and then evaporated with DCM. An orange oil was obtained. Yield: 120 mg, 65%. The product was obtained as a mixture of diastereoisomers. 1H NMR (400MHz, CDCl3) δ8.30(s, 1H), 7.33-7.22(m, 1H), 7.22-7.06(m, 1H), 6.85(d, J=8. 6Hz, 1H), 6.73-6.64(m, 1H), 6.45-6.13(m, 1H), 6.13-5.98(m, 1H), 5.48-5.40(m, 1H), 5 .35-5.30(m, 1H), 5.20-4.88(m, 2H), 4.82-4.65(m, 3H), 4.30-4.22(m, 2H), 3.05-2.85 (m, 6H), 1.95-1.85(m, 4H), 1.55-1.38(m, 14H), 1.35-1.25(m, 4H), 0.92-0.75(m, 16H). m / z 735.2[M+H] +

[0081] Stage 2: [ka] A solution of ethyl(9S,12S,E)-6-(2-(3-(((allyloxy)carbonyl)amino)-4-chlorophenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-oate (91 mg, 1 eq, 124 μmol) was bubbled with nitrogen in tetrahydrofuran (6.0 mL). Pd(PPh3)4 (7.1 mg, 0.05 eq, 6.19 μmol) was added, followed by the addition of tri-n-butyltin hydride (72.1 mg, 66.1 μL, 2 eq, 248 μmol). The mixture was stirred under a nitrogen atmosphere at room temperature for 2 hours. The solvent was removed under reduced pressure, and the crude substance was purified by flash column chromatography (12 g SiO2, 0-10% methanol in dichloromethane). The fractions were combined and concentrated to obtain 79 mg of orange oil. This was purified by flash column chromatography (4 g SiO2, 0-40% ethyl acetate in heptane) to obtain the product as a white solid. Yield: 53.2 mg, 66%. 1H NMR (400MHz, CDCl3) δ7.08-6.62(m, 4H), 6.35-6.05(m, 1H), 5.16-5.05(m, 1H), 4.65(dd, J=39.1, 9.2Hz, 1H), 4.30- 4.20(m, 2H), 3.93-3.78(m, 2H), 3.05-2.88(m, 6H), 1.96(d, J=10.1Hz, 4H), 1.61-1.28(m, 19H), 0.95-0.72(m, 16H). m / z 651.2[M+H] +

[0082] Stage 3: [ka] A solution of lithium hydroxide monohydrate (16.4 mg, 5 eq, 390 μmol) in water (1.0 mL) was added to a solution of ethyl (9S,12S,E)-6-(2-(3-amino-4-chlorophenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-oate (51 mg, 1 eq, 78.1 μmol) in methanol (2.0 mL). The solution turned yellow. The reaction mixture was stirred overnight at room temperature. After neutralizing the reaction mixture with acetic acid, it was extracted with ethyl acetate (3x). The organic layer was washed with brine, dried over sodium sulfate, concentrated, and co-evaporated with dichloromethane (2x) to obtain a beige solid. Yield: 46 mg, 95% m / z 623.2[M+H] +

[0083] Stage 4: [ka] To a solution of (9S,12S,E)-6-(2-(3-amino-4-chlorophenyl)propan-2-yl)-9-(tert-butyl)-12-isopropyl-2,2,5,11,14-pentamethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadec-13-en-15-acid (44 mg, 1 eq, 74 μmol) in dichloromethane (1.0 mL), 10% TFA in dichloromethane (1.0 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. The mixture was cooled to 0°C, and TFA (740 mg, 500 μL, 87 eq, 6.49 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, and the residue was purified using acidic preparative MPLC (Luna 5-40). The product fractions were combined and then freeze-dried. Yield: 17.8 mg, 48%. The product was obtained as a mixture of diastereoisomers. The diastereoisomers were separated by RP-HPLC to produce S,S,S (5.5 mg) and R,S,S (7.5 mg) isomers. 1 H NMR (400MHz, DMSO-d6) δ7.82(s, 1H), 6.83-6.61(m, 4H), 5.40-4.10(m, 4H), 3.38(s, 2H), 3.24-2.96(m, 4H) ), 2.20-1.94(m, 4H), 1.84(d, J=1.4Hz, 3H), 1.28-1.13(m, 6H), 0.92(d, J=24.6Hz, 9H), 0.85-0.68(m, 6H). m / z 523.4[M+H] +

[0084] [Example 4] Analysis of cell viability FaDu cell line or A549 cell line was seeded at 3000 cells per well in a 96-well plate and incubated at incubator temperature (37°C, 5% CO2). After 24 hours, 100 μl of drug at nine different concentrations (serially diluted in 1 / 5 increments starting from 1000 nM) was administered to the cell lines. The drugs used were the compound of chemical formula 2 prepared in Example 1, SC209 (MedChemExpress), and MMAE. A control group (drug concentration 0) was also prepared without drug treatment. After incubation at incubator temperature (37°C, 5% CO2) for 3 days, 100 μl of CellTiter-Glo reagent (using CellTiter-Glo® Luminescent Cell Viability Assay kit (Promega, G7571)) was added to each well of the plate and then pipetted. After 10 minutes of insothermal incubation (RT), luminescence was measured. If the luminescence value at a drug concentration of 0 was set to 100%, then the concentration that showed 50% of that luminescence value was the IC (Impulse Coefficient). 50 It is a value.

[0085] [Table 1]

[0086] The results are shown in Figures 1 and 2 and Table 1. Referring to these results, it can be confirmed that when lung cancer cell lines (A529) and head and neck squamous cell carcinoma cell lines (FaDu) are treated with the compound of the present invention, the cell viability is reduced to a similar level to that achieved with SC209 and MMAE. That is, the compound of the present invention (chemical formula 2 in this example) has a cancer cell death effect. As confirmed from the above examples, the compound according to the present invention exhibits a cancer cell death effect and can therefore be used as a cancer prevention or treatment composition containing the compound.

[0087] While not particularly limited herein, the compounds may be provided in forms of drugs commonly used in the art, in addition to compositions containing the compounds alone. For example, the compounds of the present invention may be provided in the form of drug-linkers, in which the compounds are conjugated as a drug with a linker; drug-carriers, in which the compounds are conjugated as a drug with a carrier; or drug-linker-carriers (where the carrier includes antibodies, aptamers, repebodies, etc.). More specifically, drugs in the form of antibody-drug conjugates containing the compounds as a payload can be provided, which can be used in a variety of ways, such as in compositions for co-administration. It is obvious to the ordinary person in the art that the drug-linker, drug-carrier, or drug-linker-carrier forms all have cell-killing effects equivalent to or similar to those of the compounds of the present invention alone, and complexes in which the compounds of the present invention are conjugated with known peptide-based linkers such as GGFG and val-cit, or acid-sensitive linkers such as CL2A, also fall within the scope of the present invention.

[0088] The above description focuses on preferred embodiments of the present invention. Those with ordinary skill in the art to which the present invention pertains will understand that the present invention can be embodied in modified forms that do not depart from its essential characteristics. Therefore, the disclosed embodiments should be considered in an explanatory rather than restrictive manner. The scope of the present invention is defined in the claims, not in the foregoing description, and all differences within an equivalent scope should be construed as being included within the scope of the present invention.

Claims

1. Compounds of chemical formula 1, their derivatives, their isomers, their pharmaceutically acceptable salts or solvates: [Chemical formula 1] 【Chemistry 1】 In the above chemical formula 1, R is selected from halogen, hydroxyl, carboxyl group, C1-C6 alkyl, and C1-C6 alkoxy.

2. The derivative of the chemical formula 1 is a compound represented by the following chemical formula 2 or 3, the compound according to claim 1, its isomer, its pharmaceutically acceptable salt or solvate: [Chemical formula 2] 【Chemistry 2】 [Chemical formula 3] 【Transformation 3】

3. The derivative of the chemical formula 1 is a compound represented by the following chemical formula 4 or 5, the compound according to claim 1, its isomer, its pharmaceutically acceptable salt or solvate: [Chemical formula 4] 【Chemistry 4】 [Chemical formula 5] 【Transformation 5】

4. The derivative of the chemical formula 1 is a compound represented by the following chemical formula 6 or 7, the compound according to claim 1, its isomer, its pharmaceutically acceptable salt or solvate: [Chemical formula 6] 【Transformation 6】 [Chemical formula 7] 【Transformation 7】

5. A carrier-drug complex comprising a compound of chemical formula 1, its derivatives, its isomers, its pharmaceutically acceptable salts or solvates; and a carrier linked to the compound: [Chemical formula 1] 【Transformation 8】 In the above chemical formula 1, R is selected from halogen, hydroxyl, carboxyl group, C1-C6 alkyl, and C1-C6 alkoxy.

6. The carrier drug complex according to claim 5, wherein the derivative of chemical formula 1 is a compound represented by the following chemical formula 2 or 3: [Chemical formula 2] 【Chemistry 9】 [Chemical formula 3] 【Chemistry 10】

7. The carrier drug complex according to claim 5, wherein the derivative of chemical formula 1 is a compound represented by the following chemical formula 4 or 5: [Chemical formula 4] 【Chemistry 11】 [Chemical formula 5] 【Chemistry 12】

8. The carrier drug complex according to claim 5, wherein the derivative of chemical formula 1 is a compound represented by the following chemical formula 6 or 7: [Chemical formula 6] 【Chemistry 13】 [Chemical formula 7] 【Chemistry 14】

9. The carrier-drug complex according to claim 5, wherein the carrier-drug complex is in a form in which the carrier is conjugated with a drug via a linker.

10. The carrier drug complex according to claim 5, wherein the linker comprises GGFG or val-cit.

11. The carrier-drug conjugate according to claim 8, wherein the carrier is an antibody, peptide, repebody, or aptamer.

12. The antibodies mentioned above are urelumab, utomirumab, bebuterovimab, aducanumab, bapineuzumab, crenezumab, donanemab, gantenerumab, lecanemab, solanezumab, nesvacumab, evinacumab, and enobrituzumab. blituzumab, omburtamab, belimumab, ianalumab, tabalumab, bertilimumab, mogamulizumab, leronlimab, ciplizumab, foralumab, muromonab-CD3, otelixizumab, teplizumab ), ibalizumab, tregalizumab, zanolimmab, itolizumab, efalizumab, inebilizumab, tafasitamab, tositumomab, ocrelizumab, ofatumumab, rituximab, ublituximab, beltz Veltuzumab, Epratuzumab, Basiliximab, Daclizumab, Vallilumab, Lulizumab, Iratumumab, Lintuzumab, Daratumumab, Felzartamab, Isatuximab, Mezagitamab, BleselumabDacetuzumab, Iscalimab, Lucatumumab, Mitazalimab, Sotigalimab, Dapirolizumab, Apamistamab, Ligufalimab, Magrolimab, Alemtuzumab, Crizanlizumab, Inclacumab b) Cusatuzumab, Oleculumab, Miratuzumab, Galiximab, Carotuximab, Adecatumumab, Eptinezumab, Erenumab, Fremanezumab, Galcanezumab, Zolbetuximab, Onartuzumab ), eculizumab, pozelimab, ravulizumab, lacnotuzumab, axatilimab, cabilalizumab, emactuzumab, ipilimumab, quavonlimab, tremelimumab, zalifrelimab, cetuximab Depatuxizumab, Futuximab, Imgatuzumab, Matuzumab, Modotuximab, Necitumumab, Nimotuzumab, Panitumumab, Tomuzotuximab, Zalutumumab, Batoclimab, Nipocalimab,Rozanolixizumab, Burosumab, Farletuzumab, Dinutuximab, Naxitamab, Ragifilimab, Gimsilumab, Lenzilumab, Mavrilimumab, Namilumab, Otilimab, Pronmarlimab, Kodori Tuzumab (Codrituzumab), Margetuximab, Pertuzumab, Trastuzumab, Datopotamab, Patritumab, Seribantumab, Dunogotuzumab, Ficlatuzumab, Rilotumumab, Aromfilimab, Aniflorumab olumab, emapalumab, ligerizumab, omalizumab, cyclostomab, dalotuzumab, figitumumab, ganitumab, teprotumumab, bermekimab, canakinumab, gevokizumab, briakinumab ), ustekinumab, anlukinzumab, sendakimab, lebrikizumab, tralokinumab, brodalumab, bimekizumab, ixekizumab, secukinumab, brazikumab, guselkumab, mirikizumab,Risankizumab, Tildrakizumab, Nemolizumab, Imsidolimab, Spesolimab, Pascolizumab, Dupilumab, Depemokimab, Mepolizumab, Reslizumab, Benralizumab, Clazakizumab ), olokizumab, siltuximab, silukumab, ziltivekimab, levirimab, sarilumab, satralizumab, tocilizumab, abituzumab, favezerimab, fianlimab, ieramilimab, relatrimab (Rel) atlimab, simtuzumab, avagovomab, oregovomab, tanezumab, ivuxolimab, locatinlimab, tavolimab, telazololimab, vonlerolizumab, alirocumab, bococizumab, ebronucimab ), evolocumab, flubocimab, ongericimab, taforecimab, dostarlimab, valstilimab, camrelizumab, cemiplimab, geptanolimab, nivolumab, pembrolizumab, penplimab,Pidilizumab, Progolimab, Retifanlimab, Sasanlimab, Serplulimab, Sintilimab, Spartalizumab, Tislerizumab, Tripalimab, Ezabenlizumab, Zimberelimab, Atezolizumab lizumab, avelumab, cosibelimab, sugemalimab, durvalumab, envafolimab, subratoxumab, denosumab, zirovertamab, elotuzumab, domvanalimab, etigilimab, osperirumab b) Tiragolumab, Vibostolimab, Surzebiclimab, Cobolimab, Sabatolimab, Concizumab, Marstacimab, Adalimumab, Golimumab, Infliximab, Certolizumab, Conatumumab, Gatuzumab, Tezeperumab, Gatipotuzumab, Cabilizumab, Bevacizumab, Brolucizumab, Ranibizumab, Olinvacimab, Icrucumab, Ramucirumab, Caplacizumab, Abrilumab,Etrolizumab, Vedolizumab, Intetumumab, Natalizumab, Obrindatamab, Elranatamab, Limboseltamab, Teclistamab, Epcoritamab amab), Glofitamab, Mosunetuzumab, Odronextamab, Flotetuzumab, Vibecotamab, Catumaxomab, Cibisatamab, Talquetamab, Ubamatamab, Emfizatamab, Blinatumomab, Amibanta Amivantamab, Emicizumab, Xenocutuzumab, Zanidatamab, Tibrizumab, Naptumomab, Belantamab, Pivekimab, Praluzatamab, Coltuximab, Denintuzumab, Loncastuximab, Ibritumoma Ibritumob, Inotuzumab, Epratuzumab, Moxetumomab, Brentuximab, Gemtuzumab, Vadastuximab, Lorbotuzumab, Polatuzumab, Tusamitamab, Terisotuzumab, Robalpituzumab ab), Depatuxizumab, Farletuzumab, Mirbetuximab, Disitamab, Anetumab, Enfortumab, Sacituzumab, Vobarilizumab, Cadonilimab, Vudalimab, Teboterimab, Ivonescimab,The carrier drug conjugate according to claim 8, wherein the conjugate is at least one selected from the group consisting of erfonrilimab, ozoralizumab, faricimab, vanucizumab, and navicixizumab.

13. A drug-linker comprising a compound of chemical formula 1, its derivatives, its isomers, its pharmaceutically acceptable salts or solvates; and a carrier linked to the compound: [Chemical formula 1] 【Chemistry 15】 In the above chemical formula 1, R is selected from halogen, hydroxyl, carboxyl group, C1-C6 alkyl, and C1-C6 alkoxy.

14. The drug-linker according to claim 13, wherein the linker comprises GGFG or val-cit.

15. Pharmaceutical compositions for the prevention or treatment of cancer, comprising a compound of chemical formula 1, its derivatives, its isomers, its pharmaceutically acceptable salts or solvates: [Chemical formula 1] 【Chemistry 16】 In the above chemical formula 1, R is selected from halogen, hydroxyl, carboxyl group, C1-C6 alkyl, and C1-C6 alkoxy.

16. The pharmaceutical composition according to claim 15, wherein the compound of chemical formula 1 is linked to a carrier via a linker.

17. The aforementioned cancers include pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, epithelial ovarian cancer, ovarian germ cell tumor, male breast cancer, brain tumor, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, nasal cavity cancer / paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain tumor, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, Renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue tumors, malignant bone tumors, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary origin, gastric lymphoma, stomach cancer, gastric carcinoid tumor, gastrointestinal stromal tumor, Wilms' tumor, breast cancer, triple-negative breast cancer (TNBC), sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer The pharmaceutical composition according to claim 15, which is at least one selected from the group consisting of adenocarcinoma, metastatic bone tumor, metastatic brain tumor, mediastinal tumor, rectal cancer, rectal carcinoid tumor, vaginal cancer, spinal cord cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, hematological cancer, and thymic cancer.

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