Compound comprising self-destructing group and ligand-drug conjugate comprising same

Abstract

The present invention provides a compound comprising a self-destructing group and a ligand-drug conjugate comprising the same. According to the present invention, the active substance can be stably delivered to the target site, and the active ingredient can be rapidly released at the target site, thereby increasing the efficacy of the active substance and inhibiting the active substance from causing side effects at positions other than the target site.

Description

Technical Field

[0001] The present invention relates to a novel compound containing a self-immolative group and a ligand-drug conjugate containing the compound, and specifically to a compound containing a self-immolative group of Formula 1 and a ligand-drug conjugate of Formula 2. Background Art

[0002] Bioactive substances such as drugs and diagnostic substances have target-specific activity in the body. For example, drugs have inhibitory or therapeutic effects on specific target cells, and diagnostic substances react with specific proteins in the body to perform diagnostics.

[0003] Meanwhile, since bioactive substances may be toxic to biological materials other than the target, technologies have been proposed that suppress the side effects of drugs while allowing the drug effects to appear selectively on target cells.

[0004] For example, antibody-drug conjugates (ADCs) are a targeted technology that combines drugs or toxins with antibodies that bind to receptors in the body and then selectively release the drugs or toxins from target cells to exhibit the desired drug effect. Because it only releases drugs or toxins under specific conditions and selectively delivers them to target cells while minimizing side effects on normal cells, it has better efficacy than antibody therapeutics and can greatly reduce the risk of side effects.

[0005] These antibody-drug conjugates are all composed of a general "antibody-linker-drug (toxin)" structure. The linker not only simply connects the antibody and the drug, but also allows the antibody-drug conjugate to stably reach the target cells during circulation in the body. Then, the drug is separated in the target cells through antibody-drug dissociation (for example, by enzymatic hydrolysis), thereby selectively exerting its effect on the target cells. Therefore, the stability of the linker has a significant impact on the efficacy and systemic toxicity of the antibody-drug conjugate (Discovery Medicine 2010, 10(53):329-39).

[0006] Linkers for antibody-drug conjugates can generally be divided into non-cleavable and cleavable types.

[0007] Thioethers are the primary non-cleavable linkers. In cells, the bond between the drug and the linker does not dissociate, but rather between the linker and the antibody, causing the linker-bound drug to dissociate from the antibody. Thiol-maleimide linkers are commonly used, but they suffer from low chemical and plasma stability and low efficacy.

[0008] As cleavable linkers, mainly those which can be cleaved by chemical methods or hydrolyzed by enzymatic reactions are used.

[0009] Linkers with chemical dissociation mechanisms are generally composed of disulfide bonds, hydrazone bonds, or oxime bonds. However, chemically dissociative linkers may cause the drug to dissociate at sites unrelated to the target site, depending on the conditions in the blood or cells, leading to toxic side effects.

[0010] To address this issue, linkers that selectively hydrolyze within target cells via enzymatic reactions are being developed. For example, linkers that hydrolyze via enzymatic reactions do not directly connect to the drug, but rather connect via a self-immolative group (SIG) between the drug and the linker. After enzymatic hydrolysis, the drug dissociates via mechanisms such as 1,6-elimination or cyclization (Clinical Cancer Res. 2005, 11, 843-852).

[0011] However, there is still a need in the art to develop linkers that have excellent plasma stability and chemical stability, can selectively and rapidly release drugs in target cells, and have excellent versatility to form conjugates with various antibodies and drugs. Summary of the Invention

[0012] Technical issues

[0013] One object of the present invention is to provide a compound comprising a self-immolative group of Formula 1 or Formula 1-1, and a ligand-drug conjugate of Formula 2 or Formula 2-1.

[0014] Another object of the present invention is to provide a pharmaceutical composition, an imaging composition, or a detection composition comprising a compound of Formula 1 or Formula 1-1, or a ligand-drug conjugate of Formula 2 or Formula 2-1.

[0015] Solution

[0016] Each description and embodiment disclosed in this application may also be applicable to other descriptions and embodiments. In other words, all combinations of the elements disclosed in this application fall within the scope of this application. In addition, the scope of this application should not be considered to be limited by the following specific description.

[0017] In one aspect of the present invention, there is provided a compound comprising a self-immolative group represented by the following Formula 1, or a pharmaceutically acceptable salt thereof:

[0018] [Formula 1]

[0019] A-(L 1 ) k -U j

[0020] In addition, in another aspect of the present invention, a ligand-drug conjugate represented by the following Formula 2, or a pharmaceutically acceptable salt thereof is provided:

[0021] [Formula 2]

[0022]

[0023] In Formula 1 or Formula 2 of the present invention, L 1 is a divalent or multivalent linking group, k is 0 or 1, and j is 1 to 10.

[0024] In Formula 1 or Formula 2 of the present invention, A is absent, H or a binding functional group, and A' is a divalent linking group derived from A's binding functional group.

[0025] In Formula 2 of the present invention, n is a real number of 1 to 10, and E is a ligand or protein having receptor binding properties.

[0026] In Formula 1 or Formula 2 of the present invention, U is a moiety represented by the following formula A:

[0027] [Formula A]

[0028]

[0029] In formula A,

[0030] R 1 and R 2 Each independently is H or C 1-8 saturated or unsaturated hydrocarbon group;

[0031] PL is an active agent which is connected to L via a heteroatom selected from N, O and S. 2 or with R 1 and R 2 The bonded carbon atoms are connected;

[0032] L 2 is a self-eliminating joint, which is selected so that L 2 With R 1 and R 2 The cleavage of the bond between the bound carbon atoms promotes L 2 Cleavage of the bond with PL;

[0033] W is an optional substituent on the benzene ring;

[0034] Z 1 and Z 3 One selected from N, NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 are each independently CH or N, and -(L 1 ) k -A and -(V) if present h Each independently replaces H in NH or CH;

[0035] Indicates that A-(L 1 ) k -bonds formed;

[0036] R 3 H or C 1-8 hydrocarbon group;

[0037] V is an electron-withdrawing group or an electron-donating group;

[0038] T is a trigger group capable of initiating the release of PL and, if present, L2, via a 1,6-elimination reaction upon cleavage;

[0039] L 3 is an optional self-immolative spacer group, which, if present, is cleaved sequentially when T is cleaved;

[0040] X and Y are each independently selected from -O-, -NH-, and S; and

[0041] h, i, l, x and y are each independently 0 or 1, and p is an integer from 0 to 2.

[0042] Formula 1 Contains self-destructive groups Compound

[0043] Contains fused ring core unit

[0044] Formula 1(A-(L 1 ) k -U j ) is represented by formula A:

[0045] [Formula A]

[0046]

[0047] In formula 1, if L 1 is a divalent linking group, then L 1 One side of the pair is bound to A, and the other side is bound to U. In this case, a U is bound to L 1 Combined. Or, if L 1 Contains branched or dendritic structures (i.e., if L 1 is a multivalent linking group), multiple U can be connected with L 1 In this case, with L 1 The number j of bound U may be from 1 to 10. In one embodiment, j is from 1 to 5. In one embodiment, j is 1.

[0048] In the above formula A, Z 1 and Z 3 One selected from N, NR 3, O, S and Se, and Z 1 and Z 3 Another one in Z 2 are each independently CH or N. -(L 1 ) k -A and -(V) if present h Each independently replaces H in NH or CH. In this case, R 3 H or C 1-8 Hydrocarbon group.

[0049] In the above formula A, connected to the Z 1 to Z 3 5-membered ring Indicates the A-(L 1 ) k -bonds formed.

[0050] In one embodiment, Z 1 and Z 3 One of the following is selected from 1 or A-bound N, NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 Can be independently connected to H, V, L 1 or A combined with C or N. R 3 H or C 1-8 In one embodiment, R 3 Can be H, C 1-4 Alkyl or C 1-3 alkyl.

[0051] In one embodiment, Z 1 to Z 3 Any of may contain heteroatoms selected from the group consisting of N, O, S and Se. In one embodiment, Z 1 to Z 3 Two or more of may each independently contain a heteroatom selected from the group consisting of N, O, S and Se.

[0052] In one embodiment, Z 1 and Z 3 One of them can be A or L 1 Combined N, and another and Z 2 It can be CH. Or, Z 1 and Z 3 One of the following can be selected from NR 3 , O, S and Se, and the other one and Z 2 Can be CH, or with A or L1 Combined C. Or, Z 1 and Z 3 One of the following can be selected from NR 3 , O and S, and one of the other and Z 2 Can be N, and Z 1 to Z 3 The other one can be CH, or with A or L 1 Combined C.

[0053] In one embodiment, when Z 1 to Z 3 When any one or more of is CH, V can be substituted on the carbon atom of CH. 2 When it is CH, V can replace Z 2 superior.

[0054] In one embodiment, the compound represented by Formula 1 may be a derivative of indole, benzothiophene, benzofuran, benzoselenophene, indazole, benzimidazole, benzoxazole, benzisoxazole, or benzothiazole.

[0055] In one embodiment, the ring in formula A You can select from the following groups:

[0056]

[0057] The benzene ring in the fused ring structure of formula A can be substituted by an optional substituent W, as long as it does not adversely affect the 1,6-elimination reaction triggered by the trigger group T. In one embodiment, the substituent W on the benzene ring can be selected from the group consisting of: H, C 1-12 Saturated or unsaturated hydrocarbon group, halogen, halogenated-C 1-8 Alkyl, CN, NO2, OH, C 1-8 Alkoxy, hydroxy C 1-8 Alkyl, C 1-8 Alkoxy-C 1-8 Alkyl, SH, C 1-8 Alkylthio, mercapto-C 1-8 Alkyl, amino, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino, amino-C 1-8 Alkyl, C 1-8 Monoalkylamino-C 1-8 Alkyl, C 1-8 Dialkylamino-C 1-8 Alkyl, carboxyl, C 1-8 Alkoxycarbonyl, C 1-8 Alkoxycarbonyloxy, carboxyl-C 1-8 Alkyl, C1-8 Alkoxycarbonyl-C 1-8 Alkyl, carbamoyl, mono-C 1-8 Alkylcarbamoyl, di-C 1-8 Alkylcarbamoyl, carbamoyl-C 1-8 Alkyl, mono-C 1-8 Alkylcarbamoyl-C 1-8 Alkyl and di-C 1-8 Alkylcarbamoyl-C 1-8 In Formula 1, p may be an integer from 0 to 2. In one embodiment, p may be 0.

[0058] In addition, the fused ring of formula A contains Z 1 , Z 2 and Z 3 The 5-membered ring of may be optionally substituted by an electron withdrawing group or an electron donating group V. In formula A, h is 0 or 1. In one embodiment, when A-(L 1 ) k -With Z 2 When combined, V may not exist (h is 0).

[0059] In some embodiments, V can be selected from the group consisting of halogen, CN, NO2, formyl, C 1-8 Alkylcarbonyl, carboxyl, C 1-8 Alkoxycarbonyl, carboxyl-C 1-8 Alkyl, carbamoyl, mono-C 1-8 Alkylcarbamoyl, di-C 1-8 Alkylcarbamoyl, C 1-8 Alkyl, C 1-8 Alkenyl, OH, C 1-8 Alkoxy, SH, C 1-8 Alkylthiol, NH2, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino and C6- 18 Aryl.

[0060] In some embodiments, V can be carboxyl, carboxyl-C 1-8 Alkyl, or C 1-8 Alkoxycarbonyl. For example, V can be carboxyl, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, carboxymethyl, carboxyethyl or carboxypropyl.

[0061] In formula A of the present invention, R 1 and R 2 Each independently is H or C 1-8 Saturated or unsaturated hydrocarbon group.

[0062] In one embodiment, R 1 and R2 Can be H or C independently 1-8 Alkyl or C 3-8 In one embodiment, R 1 and R 2 Can be H or C independently 1-4 Saturated hydrocarbon group, or completely or partially unsaturated hydrocarbon group. For example, R 1 and R 2 Each of them may be H. The unsaturated hydrocarbon group may include a partially unsaturated hydrocarbon group or a fully unsaturated hydrocarbon group. The hydrocarbon group may be a linear, branched or cyclic hydrocarbon group.

[0063] Self-eliminating joints

[0064] In Formula 1 of the present invention, L 2 is a self-eliminating joint, which is selected so that L 2 With R 1 and R 2 The cleavage of the bond between the bound carbon atoms promotes L 2 The bond between PL and PL can be 0 or 1. When l is 0, PL can directly bind to R 1 and R 2 The bound carbon atoms are bound.

[0065] When the compound of Formula 1 comprises a self-immolative linker, L 2 The group is removed from R by a 1,6-elimination reaction triggered by a trigger group T. 1 and R 2 The bound carbon atom is cleaved, and PL - Or PL-H can be obtained from L 2 Released on.

[0066] In one embodiment, L 2 is at least one selected from -OC(=O)-, -S(=O)2-, In this case, R 10 to R 12 Can be H, C 1-8 Alkyl, amino-C 1-8 Alkyl, mono-(C 1-8 alkyl)amino or di-(C 1-8 Alkyl)amino substituted C 1-8 Alkyl, or -(CH2CH2O) g R 13 In this case, R 13 Can be H or C 1-4 Alkyl, g can be an integer from 1 to 10. In one embodiment, C 1-8 The alkyl group can be C 1-6Alkyl, C 1-4 Alkyl or C 1-3 Alkyl. For example, R 10 to R 12 Each independently may include H, methyl, ethyl, propyl, 2-aminoethyl, 2-(N-methylamino)ethyl, 2-(N,N-dimethylamino)ethyl, 2-(N-ethylamino)ethyl, or 2-(N,N-diethylamino)ethyl. In one embodiment, the moiety of formula A may include -OC(=O)- as L 2 .

[0067] Surfactant (PL)

[0068] In formula A of the present invention, PL is an active agent that exhibits a desired biological activity in target cells, which is formed by reacting a heteroatom selected from N, O and S in the active agent with L. 2 or with R 1 and R 2 When T is cleaved by an enzymatic reaction or a chemical reaction, PL is released from the compound of Formula 1 through a 1,6-elimination reaction.

[0069] In one embodiment, PL can be at least one active agent selected from the group consisting of drugs, toxins, fluorophores, affinity ligands, diagnostic substances and detection probes. Specifically, the heteroatoms (selected from N, O and S) contained in drugs, toxins, fluorophores, affinity ligands, diagnostic substances and detection probes can be combined with R 1 and R 2 Therefore, as long as it contains a carbon atom selected from N, O and S that can be combined with R 1 and R 2 The bound carbon atom or L 2 (If L 2 Any heteroatom (e.g., present or present) bound to a tertiary amine group, or a functional group containing such a heteroatom may be additionally introduced therein, may serve as the PL of the present invention. It should be noted that the PL of the present invention is not limited to the specific active agents exemplified herein. For example, the PL may include a functional group in which the H atoms of a primary or secondary amine group, a hydroxyl group, or a carboxyl group have been removed, a functional group in which the lone pair of electrons of the nitrogen atom of a tertiary amine group has been donated, or a functional group bonded to a nitrogen atom via an imine addition reaction.

[0070] The drug can be selected from the group consisting of, but not limited to, erlotinib (TARCEVA; Genentech / OSIPharm.); bortezomib (VELCADE; MilleniumPharm); fulvestrant (FASLODEX; AstraZeneca); sutent (SU11248; Pfizer); letrozole (FEMARA; Novartis); imatinib mesylate (GLEEVEC; Novartis); PTK787 / ZK222584 (Novartis); oxaliplatin (Eloxatin; Sanofi); 5-fluorouracil (5-FU); folinic acid; rapamycin (Sirolimus, RAPAMUNE; Wyeth); lapatinib (TYKERB, GSK572016; GlaxoSmithKline); lonafarnib (SCH66336); sorafenib (BAY43-9006; Bayer Labs.); gefitinib (IRESSA; Astrazeneca); AG1478, AG1571 (SU5271; Sugen); alkylating agents (e.g., thiotepa or cyclophosphamide); alkyl sulfonates (e.g., busulfan, improsulfan, or zinsulfanil); aziridines (e.g., benzodopa, carboquinone, metodepa, or uredepa); ethyleneimine, methylmelamine, hexamethylmelamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphamide, trihydroxymethylmelamine; acetogenins (e.g., balataxine or balataxine ketone); camptothecins, including synthetic analogs of topotecan; bryostatin; callystatin; CC-1065 (including its synthetic analogs of adolesine, carzelesin, or biszelesin); cryptophycins (e.g., cryptophycin 1 or cryptophycin 8); dolastatin; duocarmycins cin) (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; panratistatin; sarcodictyin; spongistatin; nitrogen mustards (e.g., chlorambucil, naphthiazolin, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, or uracil mustard); nitrosoureas (e.g., carmustine, chlorozolin, fotemustine, lomustine, nimustine, or ranimnustine); antibiotics (e.g., enediyne antibiotics selected from calicheamicin gamma 1 I and calicheamicin ωI1, or dynemicin, including dynemicin A); bisphosphonates (e.g., clodronate); esperamicin, neosporin chromophore, or related chromoprotein enediyne antibiotic chromophores, aclacinomycins, actinomycins, anramycins, azaserine, bleomycins, cactinomycins, carabicin, carninomycin, carbomycin, chromomycin, dactinomycin, daunorubicin, detorubucin, 6-diazo-5-oxo-L-norleucine, ADRLIMYCIN;Adriamycin (e.g., morpholinodriamycin, cyanomorpholinodriamycin, 2-pyrroline-driamycin, liposomal doxorubicin, or deoxydriamycin), epirubicin, esorubicin, mexilomycin, mitomycin (e.g., mitomycin C, mycophenolic acid, nogamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptomycin, streptozocin, tuberculin, ubenimex, zinostatin, or zorubicin); antimetabolites (e.g., 5-fluorouracil (5-FU)); folic acid analogs (e.g., leucovorin, methotrexate, pteroxatin, or trimetrexate); purine analogs (e.g., fludarabine, 6-fluorouracil, 5 ... -mercaptopurine, thioetherpurine, or thioguanine); pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, or floxuridine); androgens (e.g., captestosterone, drostanolone propionate, cyclothiosteroid, melastane, or testolactone); antiadrenal agents (e.g., aminoglutethimide, mitotane, or trilostane); folic acid supplements (e.g., folinic acid); aceglucuronolide; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; anthracene; edatraxate; defofamine; demequinoxaline; diaziquone; elfornithine; elliptinium acetate); epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansine-like substances (e.g., maytansine or ansamitocin); trichothecenes (e.g., T-2 toxin, verracurin A, baculoanthrene A, or anguidine); mitoxantrone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; methylprocarbazine; Polysaccharides; razoxane; rhizoxin; sizolan; spirogermanamine; tenuazonic acid; triazinon; 2,2',2"-trichlorotriethylamine; trichothecenes (particularly, T-2 toxin, verracurin A, baculocin A, and anguidine); urethanes; vindesine; dacarbazine; mannitol mustard; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; cytarabine ('Ara-C'); cyclophosphamide; thiotepa; taxanes (e.g., Paclitaxel (TAXOL; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE™ non-hydrogenated castor oil, albumin-engineered nanoparticle paclitaxel formulation (American Pharmaceutical Partners, Schaumber, I11), or TAXOTERE; docetaxel (TAXOTERE); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; platinum analogs (e.g., cisplatin or carboplatin); vinblastine; platinum; etoposide, ifosfamide; mitoxantrone; vincristine; vinorelbine (NAVELBINE); mitoxantrone; teniposide; edatrexate; daunorubicin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DFMO); retinoids (e.g., retinoic acid); capecitabine, and pharmaceutically acceptable salts, solvates, acids or derivatives thereof.

[0071] Other drugs in addition to the above drugs include, but are not limited to: (i) antihormonal agents that modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including: tamoxifen (NOLVADEX, including tamoxifen), raloxifene, droloxifene, 4-hydroxytamoxifen, trovaxifen, raloxifene, LY117018, onapristone and FAREATON; toremifene; (ii) aromatase inhibitors that inhibit aromatase to regulate estrogen production in the adrenal glands, such as 4(5)-imidazole, aminoglutethimide, MEGASE; megestrol acetate, AROMASIN; exemestane, FEMARA; letrozole and ARIMIDEX; anastrozole; (iii) antiandrogen agents, such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog) ); (iv) aromatase inhibitors; (v) protein kinase inhibitors; (vi) lipid kinase inhibitors; (vii) antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in cell adhesion, for example, PKC-α, Raf, H-Ras; (viii) ribozymes, for example, VEGF inhibitors, such as ANGIOZYME ribozyme and HER2 expression inhibitors; (ix) vaccines, such as gene therapy vaccines; ALLOVECTIN; vaccine, LEUVECTIN vaccine and VAXID vaccine; PROLEUKIN; rlL-2; LURTOTECAN; topoisomerase 1 inhibitors; ABARELIX; rmRH; (x) anti-angiogenic agents, such as bevacizumab (AVASTIN, Genentech); and (xi) pharmaceutically acceptable salts, solvates, acids or derivatives thereof.

[0072] In one embodiment, the drug can be selected from a cytokine, an immunomodulatory compound, an anticancer agent, an antiviral agent, an antibacterial agent, an antifungal agent, an anthelmintic, or a combination thereof.

[0073] Cytokines are small protein molecules secreted by many cells for cell signaling, which can be signaling molecules widely used for intercellular communication. Cytokines include monocytokines, lymphokines, traditional polypeptide hormones, etc. Exemplary cytokines can include, but are not limited to, growth hormone (e.g., human growth hormone, N-methionyl human growth hormone, or bovine growth hormone); parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones (e.g., follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), or luteinizing hormone (LH)); hepatocyte growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor α; tumor necrosis factor β; Mullerian inhibitory substance; mouse gonadotropin-related peptide; inhibin; activin; vascular endothelial growth factor; integrin, thrombopoietin (TPO); nerve growth factor (e.g., NGF-β); platelet growth factor; transforming growth factor (TGF) (e.g., TGF-α or TGF-β ); insulin-like growth factor-I, insulin-like growth factor-II; erythropoietin (EPO); osteoinductive factors; interferons (e.g., interferon alpha, interferon beta, or interferon gamma); colony stimulating factor (CSF) (e.g., macrophage CSF (M-CSF), granulocyte-macrophage CSF (GM-CSF), or granulocyte-CSF (G-CSF)); interleukins (IL) (e.g., IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, or IL-12); tumor necrosis factor (e.g., TNF-α or TNF-β); and polypeptide factors (e.g., LIF or kit ligand (KL)). In addition, the term "cytokine" also includes proteins from natural sources or recombinant cell culture and biologically active equivalents of native sequence cytokines.

[0074] The immunomodulatory compound can be selected from the group consisting of aminocaproic acid, azathioprine, bromocriptine, chloroquine, chlorambucil, cyclosporine, cyclosporin A, danazol, DHEA (dehydroepiandrosterone), dexamethasone, etanercept, hydroxychloroquine, hydrocortisone, infliximab, meloxicam, methotrexate, cyclophosphamide, mycophenolate mofetil, prednisone, sirolimus and tacrolimus.

[0075] The anticancer agent may be selected from the group consisting of methotrexate, paclitaxel, L-asparaginase, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosoureas, cisplatin, carboplatin, mitomycin, dacarbazine, procarbazine, topotecan, nitrogen mustards, carbazine, etoposide, 5-fluorouracil, BCNU (bis-chloroethyl nitrosourea), irinotecan, camptothecin, exitecan, belotecan, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone , asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, chlorambucil, melphalan, carmustine, lolimus, busulfan, sulphonate, dacarba, etoposide, teniposide, topotecan, 9-aminocamptothecin, crestofop, mitomycin C, trimetrexate, mycophenolic acid, thiazofurine, ribavirin, EICAR (5-ethynyl-1-β-D-ribofuranose imidazolyl-4-carboxamide), hydroxyurea, deferoxamine, floxuridine, doxifluridine, raltitrexed, cytarabine (arabinoside) C), cytosine arabinoside, fludarabine, tamoxifen, raloxifene, megestrol acetate, goserelin, leuprorelin acetate, flutamide, bicalutamide, EB1089, CB1093, KH1060, verteporfin, phthalocyanine, photosensitizer Pe4, demethoxyhypocrellin A, interferon-α, interferon-γ, tumor necrosis factor, gemcitabine, Velcade, revamid, thalamid, lovastatin, 1-methyl-4-phenylpyridinium ion, staurosporine, actinomycin D, dactinomycin, bleomycin A2, bleomycin B2, peplomycin, epirubicin, pirarubicin, daunorubicin, mitoxantrone, verapamil, and thapsigargin. The antiviral agent can be selected from the group consisting of: pencicyclovir, valacyclovir, gancicyclovir, foscarnet, ribavirin, idoxuridine, vidarabine, trifluridine, acyclovir, famcicyclovir, amantadine, rimantadine, cidofovir, antisense oligonucleotides, immunoglobulins and interferons. The antibacterial agent can be selected from the group consisting of: chloramphenicol, vancomycin, metronidazole, trimethoprim, sulfamethoxazole, quinupristin, dalfopristin, rifampicin, spectinomycin and nitrofurantoin.The antifungal agent may be selected from the group consisting of amphotericin B, candidatin, filipin, hamycin, natamycin, nystatin, cytomycin, bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omeprazole, oxiconazole, silaconazole, sulconazole, tioconazole, albaconazole, fluconazole, isavuconazole, itraconazole, posaconazole, ravuconazole, terconazole, voriconazole, abafungin, amorolfin, butenafine, naftifine, terbinafine, anidulafungin, caspofungin, micafungin, benzoic acid, ciclopirox, flucytosine, griseofulvin, haloprogin, tolnaftate, undecylenic acid, crystal violet, balsam of Peru, ciclopirox olamine, piroctone olamine, zinc pyrithione, and selenium sulfide. The anthelmintic may be selected from the group consisting of mebendazole, pyrantel pamoate, thiabendazole, diethylcarbamazine, ivermectin, niclosamide, praziquantel, albendazole, rifampicin, amphotericin B, melarsoprol, eflornithine, metronidazole, tinidazole and miltefosine.

[0076] "Toxin" refers to a toxic substance produced within living cells or organisms. Toxins can be small molecules, peptides, or proteins that can interact with biological macromolecules (e.g., enzymes or cell receptors) upon contact with or absorption by body tissues, causing disease. Furthermore, "toxin" includes plant toxins and animal toxins. Exemplary animal toxins include, but are not limited to, diphtheria toxin, botulinum toxin, tetanus toxin, dysentery toxin, cholera toxin, tetrodotoxin, dinoflagellate toxin, and ciguatera toxin. Exemplary plant toxins include, but are not limited to, ricin and AM toxin.

[0077] For example, small molecule toxins may include, but are not limited to, auristatins, tubulysin, geldanamycin (Kerr et al., 1997, Bioconjugate Chem. 8(6):781-784), maytansinoids (EP1391213, ACR2008, 41, 98-107), calicheamicin (US2009105461, Cancer Res. 1993, 53, 3336-3342), daunomycin, doxorubicin, methotrexate, vinblastine, SG2285 (Cancer Res. 2010, 70(17), 6849-6858), dolastatin, dolastatin analogs auristatin (US5635483), cryptophycin, camptothecin, root toxin derivatives, CC-1065 analogs or derivatives, duocarmycin, enediyne antibiotics, esperamicins, epothilones, PBD (pyrrolobenzodiazepine) derivatives, α-amanitin and toxinoids. Toxins can exhibit cytotoxic and cell growth inhibitory activities through tubulin binding, DNA binding and topoisomerase inhibition.

[0078] Affinity ligands can include molecules that can form complexes with target biomolecules. Affinity ligands can be molecules that transmit signals by binding to a predetermined site on a target protein. Affinity ligands can be substrates, inhibitors, stimulants, neurotransmitters, or radioisotopes.

[0079] "Detection probe" can refer to a substance or a portion of a substance that can be detected by spectral, photochemical, biochemical, immunochemical, radioactive or chemical means. For example, useful detection probes can include 32P, 35S, fluorescent dyes, electron-dense reagents, enzymes (e.g., enzymes commonly used in ELISA), biotin-streptavidin, digoxigenin, haptens and proteins for which antisera or monoclonal antibodies can be obtained, or nucleic acid molecules with sequences complementary to the target. Detection probes are generally capable of generating a measurable signal, such as a radioactive, chromogenic or fluorescent signal, which can be used to quantify the amount of the detectable portion bound in a quantitative sample. Quantification of the signal can be achieved by, for example, scintillation counting, density determination, flow cytometry, ELISA or by direct analysis of intact or subsequently digested peptides (one or more peptides can be determined) by mass spectrometry.

[0080] The above-mentioned detection probes may include (i) substances that can provide a detectable signal, (ii) substances that can change the detectable signal provided by the first probe or the second probe by causing the first probe or the second probe to react with each other (such as fluorescence resonance energy transfer (FRET)), (iii) substances that can stabilize the interaction with the antigen or ligand or increase the binding affinity, (iv) substances that can affect electrophoretic mobility or cell invasiveness through physical parameters such as charge, hydrophobicity, etc., and (v) substances that can regulate ligand affinity, antigen-antibody binding or ion complex formation.

[0081] In one embodiment, PL may be a portion of an active agent selected from the group consisting of MMAF (monomethyl auristatin F), auristatin F, MMAE (monomethyl auristatin E), SN-38, p-nitrophenol, xanthene carboxylic acid, abiraterone, gefitinib, PBD dimer, α-amanitin, seco-DUBA, doxorubicin, lapatinib, imatinib, erlotinib, exitecan, belotecan, and a compound represented by one of the following formulae:

[0082]

[0083] In one embodiment, PL may be selected from the group consisting of moieties represented by the following formula:

[0084]

[0085]

[0086] Trigger unit

[0087] In formula A of the present invention, T is a trigger group capable of initiating the release of PL, and L2 (if present), through a 1,6-elimination reaction upon cleavage.

[0088] T can be selectively cleaved in vivo through chemical or enzymatic reactions. That is, under specific conditions in vivo, T (or, if Y is present, the bond between Y and T) can be selectively cleaved, thereby stably delivering PL to the target site and selectively releasing PL at the target site.

[0089] In one embodiment, T can be connected to the benzene ring through a heteroatom such as an oxygen atom, a sulfur atom or a nitrogen atom. That is, in formula A, Y connecting the trigger group T and the compound part can be -O-, -NH- and S.

[0090] In one embodiment, -(Y) y- T can be selected from -β-galactoside, -β-glucuronide, -O-SO3 - , -NO2, -valine-citrulline derivatives, -valine-alanine derivatives, -OC(O)(CH2) r COR t1 、-O(CH2)-Ar 1 -NO2, -SC(O)(CH2) s COR t2 、-S(CH2)-Ar 2 -NO2 and -BR t3 R t4 The group composed of.

[0091] β-galactoside and β-glucuronide can be linked to the benzene ring via the oxygen atom bound to the first carbon atom of the β-galactoside moiety and the β-glucuronide moiety, and the valine-citrulline derivative and the valine-alanine derivative can be linked to the benzene ring via the nitrogen atom of the valine moiety, that is, the -β-galactoside, -β-glucuronide, -valine-citrulline derivative and -valine-alanine derivative herein can be a moiety having the following structure:

[0092]

[0093] R t1 and R t2 It can be C1-C8 alkyl, Ar 1 and Ar 2 Can be C5-C 20 Arylene or heteroarylene. t3 and R t4Each independently may be hydrogen, C1-C8 alkoxy or hydroxy, and r and s may each be an integer from 1 to 5. In one embodiment, arylene may be phenyl or naphthyl. In one embodiment, heteroarylene may contain 1 to 3 heteroatoms (N, O or S). For example, heteroarylene may be furyl, thienyl or pyrrolyl.

[0094] R t5 Can be OH, single-C 1-8 Alkylamino, di-C 1-8 Alkylamino or -NH(CH2CH2O) f R t6 , where R t6 Can be H or C 1-4 The alkyl group, f may be an integer from 1 to 10.

[0095] In one embodiment, -(Y) y -T includes: sugars such as Peptides such as and -O-SO3 - Functional groups, which can be selectively hydrolyzed by specific enzymes in the lysosome. The above-mentioned sugars may include sugars in which -OH is protected by a protecting group (e.g., acetyl) or substituted by an optional substituent. In addition, -SC(O)(CH2) s COR t2 、-S(CH2)-Ar 2 -NO 2、 -OC(O)(CH2) r COR t1 and -O(CH2)-Ar 1 -NO2 can be decomposed under reducing conditions.

[0096] For example, if Y is absent (y=0) and T is -NO2 or -BR t3 R t4 , can be cracked under reducing conditions and proton decomposition conditions respectively.

[0097] Optional self-destruct spacer unit

[0098] In formula A of the present invention, L 3 is an optional self-destructive spacer group. If it exists, it will be cleaved in sequence when T is cleaved. Under specific conditions in vivo, the 1,6-elimination reaction triggered by T cleavage makes L 3 Partial separation from the compound, so L 3 It has a structure in which the electrons generated when T is cleaved can be transferred to the benzene ring of formula A and PL.

[0099] In one embodiment, -(X) x -L3 - can be Here, R 8 and R 9 can be independently selected from H, halogen, C 1-8 The group consisting of alkyl, CN and NO2, and o can be an integer from 0 to 2. For example, -(X) x -L 3 - can be derived from benzylhydroxide (PhCH2OH) and its derivatives.

[0100] In one embodiment, Z 1 and Z 3 One of can be O, and -(X) x -L 3 - can be

[0101] In formula A of the present invention, i may be 0 or 1. In one embodiment, if i is 0, the self-immolative spacer may not exist, and -(Y)yT may be directly connected to the benzene ring.

[0102] Combination unit

[0103] In Formula 1 of the present invention, A is absent, H or a binding functional group. 1 exists (k=1), then A is H or a binding functional group, if L 1 If k does not exist (k=0), then A does not exist either.

[0104] The binding functional group may refer to, for example, a functional group that can react with a ligand or protein (E in Formula 2) or a functional group (L in Formula 1) by addition or substitution. 1 That is, the binding functional group is any functional group that can provide a ligand or protein (E) with L of formula 1. 1 Between, or (if L 1 (does not exist) a functional group for binding between U of formula 1 and another linker. In the field of ligand-drug conjugates, various functional groups for binding between ligands and linkers are known in the art. Therefore, one of ordinary skill in the art will be able to select a suitable binding functional group taking into account the structure and properties of the ligand and the linker, and it should be noted that the binding functional groups of the present invention are not limited to the specific functional groups exemplified herein. For example, the binding functional group can be bound to a ligand or protein (E) or a linker precursor having receptor binding properties through a click chemistry reaction. In one embodiment, when L in formula 1 1When it is absent (i.e., k is 0), the binding functional group further binds to the linker precursor and then can bind to the ligand or protein (E) having receptor binding properties through the functional group contained in the linker precursor. In this case, the compound of Formula 1 has an "AU" structure and the ligand-drug conjugate of Formula 2 has an "E-linker-AU" structure. In another embodiment, even if L in Formula 1 1 If present (ie, k is 1), the binding functional group can also be bound to an additional linker precursor to form an extended linker, which can then be bound to a ligand having receptor binding properties via the functional group contained in the additional linker precursor.

[0105] Therefore, when A is a binding functional group, the compound of Formula 1 of the present invention is intended to bind to an additional linker precursor or a ligand or protein (E) having receptor binding properties, and can be, for example, an intermediate for providing a ligand-drug conjugate. On the other hand, when A is absent or is H, the compound of Formula 1 of the present invention can be a complex containing an active agent (PL) that is not thought to bind to a ligand having receptor binding properties. This complex can be used for various purposes, such as changing the properties of the active agent (e.g., water solubility), targeting, etc.

[0106] In one embodiment, the binding functional group may include or consist of a functional group selected from the group consisting of halogen, OH, C 1-8 Alkoxy, hydroxyamino, COH, C 1-8 Alkylcarbonyl, carboxyl, C 1-8 Alkoxycarbonyl, tosyl, tosylate, amino, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino, NHNH2, N3, haloacetamide, maleimide, succinimide, SH, SO3H, C 1-8 Alkylsulfonyl, C 1-8 Alkoxysulfonyl, 2-pyridyl disulfide, PO3H2, OPO3H2, -N≡C, -NCS, C 4-10 Dienyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 4-10 Cycloalkynyl and C 2-8 Alkynylcarbonyl. R f can be independently H or C 1-8 In one embodiment, A can be maleimide, hydroxyamino, carboxyl, amino, N3, C 2-8 Alkynyl or For example, a maleimide group may refer to a nitrogen atom of maleimide and L 1Or U connected monovalent functional group. In one embodiment, C 1-8 The alkyl group can be C 1-6 Alkyl, C 1-4 Alkyl or C 1-3 In one embodiment, C 2-8 Alkynyl can be ethynyl, propynyl (propargyl) or butynyl.

[0107] In Formula 1 of the present invention, L 1 To connect A with formula A containing Z 1 to Z 3 A connecting group of a 5-membered ring, and k is 0 or 1.

[0108] In one embodiment, when k in Formula A is 1 and Z 1 or Z 3 When nitrogen atoms are included, L 1 Can be used with Z 1 or Z 3 In one embodiment, when k in formula A is 1 and Z 1 to Z 3 When at least one of L contains a carbon atom, 1 Can be combined with the carbon atom. For example, when Z 2 When containing carbon atoms, L 1 Can be used with Z 2 In one embodiment, AL in Formula 1 1 It can be formed by the precursor of A and L 1 The precursors of A and L 1 The conjugation between the precursors can be achieved, but is not limited to, via a bond through a click chemistry reaction, an amide bond, a urea bond, an ester bond, a carbamate bond, a disulfide bond, or a maleimide bond.

[0109] For example, the precursor of A may include at least one functional group selected from the group consisting of a hydroxyl group, an amino group, an azido group, an alkynyl group, a conjugated diene group, an alkenyl group, a cyclooctynyl group, a maleimide group, a SO2N3 group, an alkoxysulfinyl group, an oxiranyl group, an aziridine group, an oxo group, a hydrazine group, a hydroxyamino group, a thiol group, and a 1,3-dicarbonyl group. 1 The precursor of may include a chemical reaction with a precursor of A to form AL 1 Alternatively, a functional group having the structure AL 1 The linker precursor can be combined with the remainder of the compound of Formula 1.

[0110] In one embodiment, L 1 Can be C 1-200Alkylene, the middle of which chain optionally contains a divalent or multivalent functional group selected from the group consisting of amide, sulfonamide, amino, ether, carbonyl, triazole, tetrazole, sugar-derived group, sulfonated ester and dendrimer. 1-200 The alkylene group can be C 1-150 Alkylene, C 1-100 Alkylene, C 1-80 Alkylene, C 1-60 Alkylene, C 1-50 Alkylene, C 1-40 Alkylene, C 1-30 Alkylene, C 1-20 Alkylene or C 1-10 Alkylene. A sugar-derived group refers to any chemical structure formed by a covalent bond between a sugar molecule and another group. For example, a sugar-derived group may include a glycosidic bond. A dendrimer refers to an ordered three-dimensional molecular structure having branched units centered around a core. In the field of ligand-drug conjugates, various linkers of dendrimer structures are known (see, for example, Lee, et al, Nat. Biotechnol. 2005, 23, 1517-26; Almutairi, et al; Proc. Natl. Acad. Sci. 2009, 106, 685-90), which may have advantages, for example, for increasing the ratio of ligand to drug.

[0111] In one embodiment, when L 1 When it contains multivalent functional groups, that is, when L 1 When containing branched or dendritic structures, multiple U can be combined with L 1 In this case, with L 1 The number of U bound may be 1 to 10. In one embodiment, j may be 1 to 5. For example, j may be 1.

[0112] In one embodiment, when Z in Formula A 1 or Z 3 When nitrogen atoms are included, L 1 can be combined with the nitrogen atom.

[0113] In one embodiment, L 1 Can be used with Z 1 to Z 3 For example, when Z 2 When L is a carbon atom, 1 Can be used with Z 2 In one embodiment, L 1 A functional group of a -U bond (eg, carboxyl, aminocarbonyl, amino, etc.) may be specified as a substituent of the carbon atom.

[0114] In one embodiment, L1 Can include C 1-10 an alkylene group, an oxyethylene group, an amide group, a triazole ring, a tetrazole ring, an ether, a carbonyl group, or a combination thereof.

[0115] In one embodiment, L 1 May include any one selected from the group consisting of: -(CH2) na -;-(CH2CH 2O ) ma -;-(CH2OCH2) mb -;-(OCH2CH2) mc -;-C(=O)-; or a combination thereof. Wherein, R d Can be H or C 1-8 The alkyl group, na and ma to mc may each independently be an integer from 0 to 10. In one embodiment, na and ma to mc may each independently be an integer from 1 to 8, an integer from 1 to 6, or an integer from 1 to 4. When two or more types of functional groups described above are combined with each other, the order of the functional groups is not limited.

[0116] In one embodiment, in Formula 1, k may be 1, and AL 1 - may be selected from the group consisting of: N3-(CH2) n1 -;N3-(CH2CH2O) m1 -(CH2) n2 -;HO-(CH2CH2O) m1 -(CH2) n2 -;H2N-(CH2CH2O) m1 -(CH2) n2 -;H2N-O-(CH2CH2O) m1 -(CH2) n2 -;N3-(CH2CH2O) m2 -(CH2) n3 -NR d1 CO-(CH2) n4 -;R a1 NH-(CH2) n5 -;R b1 OC(=O)-(CH2) n6 -;R c1 C≡C-(CH2OCH2) m3 -CONR d2 -(CH2) n7 -; In this case, Ra1 、R b1 、R c1 、R c2 and R d1 to R d5 can be independently H or C 1-8 alkyl. In addition, n1 to n15 and m1 to m11 may each independently be an integer from 0 to 10. In one embodiment, n1 to n15 and m1 to m11 may each independently be an integer from 0 to 8, an integer from 0 to 6, or an integer from 0 to 5. In one embodiment, n1 to n15 and m1 to m11 may each independently be an integer from 1 to 8, an integer from 1 to 6, or an integer from 1 to 5.

[0117] Exemplary compounds of Formula 1

[0118] In one embodiment, the compound represented by Formula 1 of the present invention may be a compound represented by the following formula.

[0119]

[0120]

[0121] In the above formula, A, L 1 ,k,V,R 1 、R 2 、R 3 、R 11 、R 12 and PL are as described with respect to Equation 1.

[0122] The structure A-(L 1 ) k -、V、L 2 、-(X) x -L 3 -and-(Y) y The combination of -T- is only an example, and based on the embodiments described below and the disclosure of this application, compounds of Formula 1 having various combinations thereof can be easily prepared. It should be understood that all such compounds fall within the scope of this application.

[0123] In one embodiment, the compound represented by Formula 1 of the present invention may have one PL. In this case, for example, the compound represented by Formula 1 may be selected from the group consisting of the compounds shown in Table A attached hereto.

[0124] In one embodiment, DAR2-type compounds may include a functional group capable of forming a bond with a ligand (eg, an antibody) for preparing a ligand-drug conjugate, such as a maleimide functional group. Examples of these compounds are shown in the accompanying Table B.

[0125] DAR4 type compound (Formula 1-1)

[0126] In one embodiment, when the linking group L1 of Formula 1 of the present invention has a branched structure or a dendritic structure (i.e., when L1 is a multivalent linking group), a compound of Formula 1 that combines two or more active agents can be provided. In one embodiment, the compound represented by Formula 1 can be a compound represented by the following Formula 1-1.

[0127] [Formula 1-1]

[0128]

[0129] In Formula 1-1, A has the same meaning as in Formula 1.

[0130] In formula 1-1, U 1 and U 2 Each has the same meaning as U in Formula 1, wherein U 1 and U 2 They may be the same as or different from each other.

[0131] In formula 1-1, L 11 and L 12 Each has the same L as in Formula 1 1 Same meaning, where L 11 and L 12 They may be the same as or different from each other.

[0132] In Formula 1-1, j is 1 to 10. In one embodiment, j is 1 to 5. For example, j is 1.

[0133] About formula A and L 1 and U, if applicable, the contents described in Formula 1 can also be applied to A, U in Formula 1-1 respectively. 1 and U 2 and L 11 and L 12 .

[0134] In formula 1-1, L 1a and L 1b may each independently be selected from a direct bond; In this case, R e Can be H or C 1-8 alkyl.

[0135] In Formula 1-1, q1, q2, and q3 may each independently be an integer from 0 to 10. In Formula 1-1, q4 may be an integer from 1 to 10. In addition, if L 1a for Then q2 is not 0, and if L 1b for Then q3 is not 0. In one embodiment, if L 1a and / or L 1b is a direct key and q2 and q3 are 0, then L 11 and L 12 The connection structure (such as C 1-200 The alkylene group, which optionally contains a divalent or multivalent functional group selected from amide, sulfonamide, amino, ether, carbonyl, triazole, tetrazole, sugar-derived group, sulfonated ester and dendrimer in the middle of the chain, can be directly bonded to the central nitrogen atom.

[0136] In one embodiment, q1 may be an integer from 0 to 8, an integer from 1 to 8, or an integer from 1 to 6. In one embodiment, q2 and q3 may each independently be an integer from 0 to 8, an integer from 0 to 6, or an integer from 0 to 4. In one embodiment, q4 may be an integer from 1 to 8, an integer from 1 to 6, or an integer from 1 to 4.

[0137] In one embodiment, in the above formula 1-1, You can choose from the following structures: (where q2 and q3 are defined as above).

[0138] In one embodiment, A in the above formula 1-1 may include or consist of a functional group selected from the group consisting of halogen, OH, C 1-8 Alkoxy, hydroxyamino, COH, C 1-8 Alkylcarbonyl, carboxyl, C 1-8 Alkoxycarbonyl, tosyl, tosylate, amino, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino, NHNH2, N3, haloacetamide, maleimide, succinimide, SH, SO 3H 、C 1-8 Alkylsulfonyl, C 1-8 Alkoxysulfonyl, 2-pyridyl disulfide, PO3H2, OPO3H2, -N≡C, -NCS, C 4-10 Dienyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 4-10 Cycloalkynyl and C 2-8 Alkynylcarbonyl. R f can be independently H or C 1-8 For example, A in Formula 1-1 can be maleimide, hydroxyamino, carboxyl, amino, N3, C 2-8 Alkynyl or

[0139] In one embodiment, L 11 and L 12 Can be independently C 1-200 Alkylene, the middle of which chain optionally contains a divalent or multivalent functional group selected from the group consisting of amide, sulfonamide, amino, ether, carbonyl, triazole, tetrazole, sugar-derived group, sulfonated ester and dendrimer. 1-200 The alkylene group can be C 1-150 Alkylene, C 1-100 Alkylene, C 1-80 Alkylene, C 1-60 Alkylene, C 1-50 Alkylene, C 1-40 Alkylene, C 1-30 Alkylene, C 1-20 Alkylene or C 1-10 Alkylene.

[0140] In one embodiment, L 11 and L 12 Each independently may include any one selected from the group consisting of: -(CH2) na -;-(CH2CH2O) ma -;-(CH2OCH2) mb -;-(OCH2CH2) mc -;-C(=O)-; or a combination thereof. Wherein, R d , na, ma to mc are as described above with respect to Formula 1.

[0141] In one embodiment, L 11 and L 12 can be independently selected from -(CH2) n1 -;-(CH2CH2O) m1 -(CH2) n2 -;-(CH2CH2O) m2 -(CH2) n3 -NR d1 CO-(CH2) n4 -;-(CH2CH2O) m10 -(CH2) n15 -CONR d5 -(CH2) n14 -;and wherein n1, n2, n3, n4, n8, n14, and n15, and m1, m2, m8, and m10 may each independently be an integer from 1 to 8, an integer from 1 to 6, or an integer from 1 to 5. d1 and Rd5 Each independently is H or C 1-8 alkyl.

[0142] In one embodiment, the compound represented by Formula 1-1 (wherein j is 1) may be a compound represented by the following formula:

[0143]

[0144]

[0145] (In the above formula, q1 to q4, U 1 、U 2 , n1, n3, n4, n8, n14 and n15 and m2, m8 and m10 are as described above with respect to Formula 1-1. In addition, R d1 、R d5 and R e Each independently is H or C 1-8 alkyl.)

[0146] In one embodiment, the compound represented by Formula 1-1 may be a compound represented by the following formula:

[0147]

[0148]

[0149] In the above formula, q1 to q4, n1, n3, n4, n14 and n15, m2 and m10, R d1 、R d5 and R e As described above for Formula 1-1. In the above formula, PL has the same meaning as PL in Formula 1. In the above formula, Z 1 Selected from NR 3 , O, S and Se heteroatoms. 3 H or C 1-8 In the above formula, the -O-CO- group in the -O-CO-PL group is an optional self-immolative spacer group and may be absent (i.e., the -PL group is directly connected to the -CH2- group) or may be replaced by a functional group as described above with respect to the "optional self-immolative spacer group".

[0150] In one embodiment, the compound represented by Formula 1-1 can be selected from the compounds listed in Table C attached hereto.

[0151] Method for preparing compound of formula 1

[0152] Based on the examples herein and the technical knowledge of those skilled in the art of organic synthesis, the compounds of Formula 1 containing a self-immolative group of the present invention can be easily prepared by selecting appropriate solvents, starting materials, intermediates, reaction conditions, etc.

[0153] In one embodiment, the compound of Formula 1 having an indole core and a β-galactoside trigger group can be prepared according to the following Reaction Scheme 1.

[0154] [Reaction Scheme 1]

[0155]

[0156] In step 1 of reaction scheme 1, an indole starting material having a para-position of the benzene ring substituted with COH and OBn can be prepared, and the starting material is reacted with AL 1 -X (wherein X is halogen: Cl, Br, etc.) to react with AL 1 - Introducing the nitrogen atom of indole. In the reaction of step 1, an additive such as potassium carbonate may be used. Step 1 may be carried out at a temperature of 40°C to 100°C, 60°C to 100°C, or 60°C to 90°C.

[0157] In step 2, OBn can be converted to OH using boron trichloride. Step 2 can be performed at low temperatures, for example, -90°C to 0°C, -90°C to -10°C, -90°C to -20°C, -80°C to 0°C, -80°C to -10°C, or -80°C to -20°C.

[0158] In step 3, galactose in which the hydroxyl group is protected by a protecting group (-PG) can be introduced. For example, a protected galactoside group, such as acetylgalactoside group, can be introduced by reacting with a galactose into which a protecting group is introduced, such as acetylated galactose or acetyl bromide-α-D-galactose. In the reaction of step 3, additives such as benzyltributylammonium chloride, HOBt, pyridine, DIPEA, etc. can be used. In addition, additives such as silver oxide and molecular sieves can also be used. Step 3 can be carried out under the temperature conditions of -40°C to 40°C, -40°C to 30°C, -20°C to 40°C, -20°C to 30°C, -10°C to 40°C or -10°C to 30°C.

[0159] In step 4, COH connected to the benzene ring can be reduced. The reduction reaction can be carried out by a reducing agent such as sodium borohydride (NaBH4) at a low temperature of -40°C to 10°C, -40°C to 0°C, -30°C to 10°C, -30°C to 0°C, -20°C to 10°C, or -20°C to 0°C.

[0160] In step 5, a -OCO- self-eliminating linker structure can be introduced. Precursors of the -OCO- linker group can include bis(4-nitrophenyl) carbonate, 4-nitrophenyl chloroformate, and the like. Additives such as DIPEA and pyridine can be used in the reaction of step 5. Step 5 can be carried out under low temperature conditions of -40°C to 10°C, -40°C to 0°C, -30°C to 10°C, -30°C to 0°C, -20°C to 10°C, or -20°C to 0°C.

[0161] In step 6, PL can be introduced. For example, PL can be introduced by replacing a leaving group (e.g., p-nitrophenyl) attached to -OCO- with a precursor such as PL-H. In the reaction of step 6, additives such as HOBt, pyridine, and DIPEA can be used depending on the type of PL. Step 6 can be carried out at low temperatures of -40°C to 10°C, -40°C to 0°C, -30°C to 10°C, -30°C to 0°C, -20°C to 10°C, or -20°C to 0°C.

[0162] In step 7, the protecting group of the protected galactoside can be removed by hydrolysis, thereby converting it into galactoside. The hydrolysis can be carried out with an acid such as hydrochloric acid, or a base such as potassium carbonate, sodium hydroxide, or lithium hydroxide. Step 7 can be carried out at a temperature of -40°C to 40°C, -40°C to 30°C, -20°C to 40°C, -20°C to 30°C, -10°C to 40°C, or -10°C to 30°C.

[0163] Each step in Reaction Scheme 1 can be carried out in a suitable solvent selected from the group consisting of organic solvents such as methanol, DMF, MC, ACN, THF, EA and distilled water. In addition, after each step of the reaction is completed, the product can be purified by dilution, extraction and chromatography using a suitable solvent. In one embodiment, each step in Reaction Scheme 1 can be carried out under a nitrogen atmosphere.

[0164] In one embodiment, according to PL, A, L 1 and V, some steps in Reaction Scheme 1 can be omitted. For example, if PL can react with a hydroxymethyl group directly attached to a benzene ring even in the absence of -OCO- or can bind to a methyl group, Step 3 can be omitted.

[0165] Ligand-drug conjugate represented by formula 2

[0166] In one aspect of the present invention, there is provided a ligand-drug conjugate represented by the following Formula 2, or a pharmaceutically acceptable salt thereof:

[0167] [Formula 2]

[0168]

[0169] In Formula 2, E is a ligand or protein having receptor binding properties.

[0170] The compound of formula 1 according to the present invention can be combined with a ligand or protein (E) having receptor binding properties to provide a ligand-drug complex of formula 2. The ligand or protein (E) can be optionally modified to bind to the binding functional group of the compound of formula 1. In one embodiment, when the ligand E is an antibody, the moiety present at a specific position of the antibody, such as -SH of cysteine, -NH2 of lysine, -C(=O)NH2 of glutamine, -C6H4-OH of tyrosine, -SeH of selenocysteine, and -N3 and -N4 of unnatural amino acids, can be modified. It can bind to the binding functional group of the compound of formula 1. In the field of ligand-drug complexes, the types and modifications of ligand functional groups used to bind the linker-drug moiety to the ligand or protein are known in the art.

[0171] The ligand may be selected from the group consisting of peptides, tumor cell specific peptides, tumor cell specific aptamers, tumor cell specific carbohydrates, tumor cell specific monoclonal or polyclonal antibodies and antibody fragments.

[0172] In one embodiment, the protein may be selected from the group consisting of C 1-20 The group consisting of hydrocarbon groups, oligopeptides, polypeptides, antibodies, fragments of antigenic polypeptides and artificial antibodies (Repebodies). In one embodiment, the C-terminus of the protein can be a light chain or a heavy chain of an antibody.

[0173] In one embodiment, the antibody can be selected from the group consisting of intact polyclonal antibodies, intact monoclonal antibodies, antibody fragments, single-chain Fv (scFv) mutants, multispecific antibodies, bispecific antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising antigenic determinants of antibodies, and other modified immunoglobulin molecules comprising antigen recognition sites.

[0174] In one embodiment, the antibody can be selected from the group consisting of muromonab-CD3, abciximab, rituximab, daclizumab, palivizumab, infliximab, trastuzumab (Herceptin), etanercept, basiliximab, gemtuzumab, alemtuzumab, ibritumomab tiuxetan, adalimumab, afacept, omalizumab, efalizumab, tositumomab-I131, cetuximab, bevacizumab, natalizumab, ranibizumab, panitumumab, eculizumab, rilonacept, certolizumab pegol. , lindane, AMG-531 (lindane), CNTO-148 (golimumab), CNTO-1275 (ustekinumab), ABT874 (briakinumab), LEA-29Y (belatacept), belimumab, TACI-Ig (transmembrane active agent and calcium regulator and cyclophilin ligand interactor - immunoglobulin), second-generation anti-CD20, ACZ-885 (canakinumab), tocilizumab, atelizumab, mepolizumab, pertuzumab, Humax CD20 (ofatumumab), Tremelimumab (CP-675206), Tesitumumab, MDX-010 (ipilimumab), IDEC-114 (galiximab), Inotuzumab, HuMax EGFR (zalutumumab), Aflibercept (VEGF Trap-Eye), HuMax-CD4 (zalutumumab), Ala-Ala (hOKT3gamma1), Otelixizumab (ChAglyCD3; TRX4), Catumaxomab, MT-201 (adelimumab), Pregovomab, CH-14.18 (detucillinam), WXG250 (gemtuximab), AMG-162 (denosumab), AAB-001 (bapinetumab), motavizumab, MEDI524 (motavizumab), ifenguzumab, Raxicumab, third-generation anti-CD20, LY2469298 (Ocatatuzumab), and veltuzumab.

[0175] In one embodiment, the antibody can be a monoclonal antibody (mAb).

[0176] In the above formula 2, A' is a divalent linking group derived from the binding functional group (A) of formula 1. For example, A' may include a functional group formed by an addition reaction of a double bond contained in the binding functional group. In one embodiment, when the binding functional group is a maleimide group, A' may be a functional group formed by participating in an addition reaction of a double bond in the 5-membered ring of the maleimide group. If applicable, the above content regarding the binding functional group (A) of formula 1 may also apply to A'.

[0177] In one embodiment, the ligand E may be an antibody. For example, the antibody may include a functional group that binds to A in Formula 1 to form an E-A' binding structure in Formula 2. If desired, a functional group for the above-mentioned binding may be introduced into the antibody, and the types of such functional groups and methods for introducing such functional groups are known in the art.

[0178] In one embodiment, the binding structure of ligand E and A' can be represented by a portion represented by one of the following formulae. In the following formulae, * can be the remaining portion of the antibody. In the following formulae, the S, NH, CONH, C6H3-OH, Se and triazole ring moieties directly connected to * can be derived from moieties present at specific positions of ligand E, such as -SH of cysteine, -NH2 of lysine, -C(=O)NH2 of glutamine, -C6H4-OH of tyrosine, -SeH of selenocysteine, and -N3 and -NH of unnatural amino acids. part.

[0179]

[0180] In Formula 2, n may be a real number from 1 to 10. For example, n may be a real number from 1 to 6, a real number from 1 to 4, or a real number from 1 to 2.

[0181] In formula 2, U, L 1 , k and j can be compared with U and L in formula 1 1 , k and j are the same. If applicable, the above-mentioned U, L in formula 1 1 The contents of k and j can also be applied to U and L in formula 2. 1 In one embodiment, when E in the above formula 2 is an antibody and PL in U is a drug, the compound represented by formula 2 can be provided as an antibody-drug conjugate.

[0182] In one embodiment, the conjugate represented by Formula 2 may be selected from the group consisting of conjugates represented by the following formulae:

[0183]

[0184] In the above formula, mAb represents a portion of an antibody. In the above formula, m5, m6, m9, m10, n9, n10, n13, n14, and n15 can each independently be an integer from 1 to 10. In one embodiment, m5, m6, m9, m10, n9, n10, n13, n14, and n15 can each independently be an integer from 1 to 8, an integer from 1 to 6, an integer from 1 to 5, an integer from 1 to 4, or an integer from 1 to 3. R d3 and R d5 Each independently is H or C 1-8 alkyl.

[0185] In the above formula, Z 1 Selected from NR 3 , O, S and Se heteroatoms. 3 H or C 1-8 Hydrocarbon group. In the above formula, PL has the same meaning as PL in formula 1. In the above formula, the -O-CO- group in the -O-CO-PL group is an optional self-destructive spacer group, which may not exist (i.e., the -PL group is directly connected to the -CH2- group) or may be replaced by -S(=O)2-, Substitute (in the above formula, R 10 to R 12 Each independently is H, C 1-8 Alkyl, amino-C 1-8 Alkyl, mono-(C 1-8 alkyl) or di-(C 1-8 Alkyl)amino substituted C 1-8 Alkyl, or -(CH2CH2O) g R 13 , R 13 H or C 1-4 alkyl, g is an integer from 1 to 10).

[0186] In the above formula, n is a real number from 1 to 10. In one embodiment, n may be a real number from 1 to 8, a real number from 1 to 6, a real number from 1 to 4, or a real number from 1 to 2.

[0187] In one embodiment, the conjugate represented by Formula 2 may be selected from the group consisting of conjugates represented by the following formulae:

[0188]

[0189]

[0190]

[0191] In the above formula, mAb represents a portion of an antibody. In the above formula, n is a real number from 1 to 10. In one embodiment, n can be a real number from 1 to 8, a real number from 1 to 6, a real number from 1 to 4, or a real number from 1 to 2.

[0192] Exemplary DAR2-type ligand-drug conjugates

[0193] In one embodiment, the compound represented by Formula 2 can be selected from the compounds listed in the attached Table D. The specific n values ​​listed in Table D can vary in the real number range of 1 to 10. For example, n can be a real number from 1 to 8, a real number from 1 to 6, a real number from 1 to 4, or a real number from 1 to 2.

[0194] DAR4-type ligand-drug conjugate (Formula 2-1)

[0195] In one embodiment, when the linking group L of Formula 2 of the present invention 1 When having a branched or dendritic structure (i.e., when L 1 is a multivalent linking group), a ligand-drug conjugate of Formula 2 that binds multiple active agents can be provided. In one embodiment, the compound represented by Formula 2 can be a ligand-drug conjugate represented by the following Formula 2-1:

[0196] [Formula 2-1]

[0197]

[0198] In Formula 2-1, E is a ligand or protein having receptor binding properties, and A' is a divalent linking group derived from the binding functional group (A) of Formula 1. Regarding E and A', the same descriptions as in Formula 2 also apply to Formula 2-1.

[0199] In formula 2-1, U 1 and U 2 has the same meaning as U in Formula 1, wherein U 1 and U 2 can be the same as or different from each other; in addition, L 11 and L 12 With the same L as in formula 1 1 Same meaning, where L 11 and L 12 They may be the same as or different from each other.

[0200] In Formula 2-1, j is 1 to 10. In one embodiment, j is 1 to 5. For example, j is 1.

[0201] About formula A and L 1 and U, the contents described in Formula 1 can also be applied to A, U in Formula 2-1. 1 and U 2 and L 11 and L 12 In addition, regarding U 1 and U 2 and L 11 and L 12 , the contents described in Formula 1-1 can also be applied to Formula 2-1.

[0202] In formula 2-1, L 1a and L 1b may each independently be selected from a direct bond; In this case, R e Can be H or C 1-8 alkyl.

[0203] In Formula 2-1, q1, q2, and q3 may each independently be an integer from 0 to 10. In Formula 2-1, q4 may be an integer from 1 to 10. In addition, if L 1a for Then q2 is not 0, and if L 1b for Then q3 is not 0.

[0204] In one embodiment, q1 may be an integer from 0 to 8, an integer from 1 to 8, or an integer from 1 to 6. In one embodiment, q2 and q3 may each independently be an integer from 0 to 8, an integer from 0 to 6, or an integer from 0 to 4. In one embodiment, q4 may be an integer from 1 to 8, an integer from 1 to 6, or an integer from 1 to 4.

[0205] About L 1a and L 1b , q1, q2, q3, q4, the contents described in formula 1-1 can be applied to formula 2-1 respectively.

[0206] In Formula 2-1, n is a real number from 1 to 10. In one embodiment, n may be a real number from 1 to 6, a real number from 1 to 4, or a real number from 1 to 2.

[0207] In one embodiment, the ligand-drug conjugate represented by Formula 2-1 may be a conjugate represented by the following formula:

[0208]

[0209]

[0210] (In the above formula, mAb is the antibody part. In the above formula, q1 to q4, n, U 1 and U 2 、R d1 、R d5 、R e , n1, n3, n4, n8, n14 and n15, m2, m8 and m10 are as described above with respect to Formula 1-1 or Formula 2-1.)

[0211] In one embodiment, the ligand-drug conjugate represented by Formula 2-1 may be a conjugate represented by the following formula:

[0212]

[0213]

[0214] In the above formula, mAb is the antibody part. In the above formula, q1 to q4, n1, n3, n4, n14 and n15, m2 and m10, R d1 、Rd5 and R e As described above for Formula 1-1. In the above formula, PL has the same meaning as PL in Formula 1. In the above formula, Z 1 is a heteroatom selected from N, O, S and Se. In the above formula, the -O-CO- group in the -O-CO-PL group is an optional self-immolative spacer group, which may be absent (i.e., the -PL group is directly connected to the -CH2- group) or may be replaced by the functional group described above with respect to the "optional self-immolative spacer group".

[0215] In one embodiment, the ligand-drug conjugate represented by Formula 2-1 can be selected from the compounds listed in the attached Table E. The specific n values ​​listed in Table E can vary in the range of real numbers from 1 to 10. For example, n can be a real number from 1 to 6, a real number from 1 to 4, or a real number from 1 to 2.

[0216] definition

[0217] As used herein, the term "hydrocarbyl" refers to a functional group consisting of carbon and hydrogen, and refers to a saturated, partially unsaturated, or fully unsaturated straight-chain, branched, or cyclic hydrocarbon. Hydrocarbyl groups may include alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, and the like. Non-limiting examples of hydrocarbyl groups may include methyl, ethyl, propyl, butyl, ethenyl, propenyl, butenyl, ethynyl, propynyl, butynyl, and the like.

[0218] The term "alkyl" refers to a fully saturated branched or unbranched (or straight or linear) hydrocarbon. The alkyl group can be a substituted or unsubstituted alkyl group. The C1-8 alkyl group can be a C1 to C6, C1 to C5, C1 to C4, C1 to C3 or C1 to C2 alkyl group. Non-limiting examples of alkyl groups can include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl (isopentyl), neopentyl, isopentyl (iso-amyl) or n-hexyl.

[0219] The term "alkenyl" includes straight or branched alkenyl groups having 2 to 6 carbon atoms, 2 to 5 carbon atoms, or 2 to 4 carbon atoms and having one or more double bonds at any position. For example, the alkenyl group can include ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, isopentenyl, butadienyl, pentenyl, isopentenyl, pentadienyl, hexenyl, isohexenyl, hexadienyl, etc.

[0220] The term "alkynyl" refers to a straight or branched hydrocarbon chain having at least one triple bond. Alkyl is preferably a straight or branched chain having 2 to 8 carbon atoms. Examples of alkynyl may include 2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, and the like.

[0221] The term "alkoxy" refers to an alkyl group bound to an oxygen atom. For example, a C1 to C8 alkoxy group may be a C1 to C6, C1 to C5, C1 to C4, C1 to C3, or C1 to C2 alkoxy group. The alkoxy group may be a methoxy group, an ethoxy group, or a propoxy group.

[0222] The term "cycloalkyl" includes monocyclic or polycyclic saturated carbocyclic rings containing 3 to 8 carbon atoms. Examples may include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

[0223] The term "cycloalkenyl" includes non-aromatic monocyclic or polycyclic rings having 3 to 8 carbon atoms and containing at least one carbon-carbon double bond. Examples may include cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like.

[0224] The term "cycloalkynyl" refers to a monocyclic or polycyclic unsaturated hydrocarbon ring having 4 to 10 carbon atoms and containing at least one triple bond. Examples may include monocyclic alkynyls such as cyclooctynyl and cyclodecynyl. "Cycloalkynyl" in its broadest sense includes structures in which one or more carbon atoms of a hydrocarbon ring containing at least one triple bond between carbon atoms are substituted with heteroatoms (e.g., N).

[0225] The term "dienyl" refers to an unsaturated branched or unbranched C 4-10 Aliphatic substituents. Examples include, but are not limited to, 2,4-pentadienyl, 2,4-hexadienyl, 4-methyl-2,4-pentadienyl, and the like.

[0226] The term "halogen" atom refers to an atom belonging to Group 17 of the periodic table of elements. Halogen atoms include fluorine, chlorine, bromine, iodine, and the like.

[0227] The term "haloalkyl" refers to an alkyl group substituted with one or more halogen atoms.

[0228] The term "hydroxy" refers to an OH functional group (hydroxyl group).

[0229] The term "mercapto" refers to an SH functional group.

[0230] The term "cyano" is CN and refers to a functional group consisting of a triple bond between a carbon atom and a nitrogen atom.

[0231] The term "oxo" refers to =0 and "substituted by oxo" means that a carbon atom has a =0 substituent of the form -C(=O)-.

[0232] The term "nitro" refers to NO2.

[0233] The term "amino" refers to -NH2.

[0234] The term "alkylamino" refers to a functional group in which one or two hydrogen atoms of an amino group (-NH2) are replaced by one or two alkyl groups as described above, including monoalkylamino and dialkylamino groups, wherein the two alkyl groups in the dialkylamino group may be the same or different. Specifically, monoalkylamino 1-8 Alkylamino groups can be amino (-NH2) groups where one hydrogen atom is replaced by a C 1-8 Alkyl substituted, di-C 1-8 Alkylamino groups may be amino (-NH2) groups in which the two hydrogen atoms are replaced by the same or different C 1-8 Alkyl substitution. For example, mono-C 1-8 Alkylamino (-NH(C 1-8 The alkyl group may include methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, sec-butylamino, tert-butylamino, pentylamino, hexylamino, etc. 1-8 Alkylamino (-N(C 1-8 The alkyl group)2) may include, for example, dimethylamino, diethylamino, dipropylamino, methylethylamino, methylpropylamino, methylisopropylamino, methylbutylamino, methylisobutylamino, ethylpropylamino, ethylisopropylamino, ethylisobutylamino, isopropylisobutylamino, methylhexylamino, ethylhexylamino, and the like.

[0235] The term "carboxyl" refers to -COOH.

[0236] The term "carbamyl" refers to -CONH2.

[0237] The term "N-mono-C 1-8 Alkylcarbamoyl" and "N,N-di-C 1-8 "Alkylcarbamoyl" refers to a carbamoyl group (-CONH2) in which one or two hydrogen atoms bound to the nitrogen atom are replaced by C 1-8 Alkyl substituted. N,N-di-C 1-8 The two C 1-8 The alkyl groups may be the same or different.

[0238] The term "alkanoyl" refers to an alkyl group as defined above having the specified number of carbon atoms connected by a carbonyl bridge (i.e., -(C=O)-alkyl). For example, alkanoyl groups include formyl (formyl: -COH), acetyl (acetyl: -COCH3), propionyl (-COCH2CH3), butyryl (-CO(CH2)2CH3), and the like.

[0239] The term "alkanoylamino" refers to an amino group substituted with an alkanoyl group (i.e., -NH(C=O)-alkyl). The nitrogen atom of the alkanoylamino group may be substituted with other substituents, such as an alkyl group. For example, alkanoylamino groups include formylamino (-NHCOH), acetylamino (-NHCOCH3), propionylamino (-NHCOCH2CH3), butyrylamino (-NHCO(CH2)2CH3), and the like.

[0240] The term "cyano-C 1-8 Alkyl", "halogenated-C 1-8 Alkyl", "Hydroxy-C 1-8 Alkyl", "C 1-8 Alkoxy-C 1-8 Alkyl", "C 1-8 Alkoxy-C1-8alkoxy-C 1-8 Alkyl", "Carboxyl-C 1-8 Alkyl", "amino-C 1-8 Alkyl", "Carbamoyl-C 1-8 Alkyl", "N-mono-C 1-8 Alkylcarbamoyl-C 1-8 Alkyl" and "N,N-di-C 1-8 Alkylcarbamoyl-C 1-8 "Alkyl" refers to a terminal or intermediate cyano, halogen, hydroxy, alkoxy, carboxyl, amino, carbamoyl, N-mono-C 1-8 Alkylcarbamoyl and N,N-di-C 1-8 Alkylcarbamoyl-substituted C 1-8 alkyl.

[0241] The term "glycosidic group" refers to a functional group formed by a condensation reaction of a sugar molecule.

[0242] The term "heterocyclic group" refers to a saturated or partially unsaturated cyclic hydrocarbon containing at least one heteroatom. The heterocyclic group can be monocyclic or bicyclic. The bicyclic heterocyclic group can be a spirocyclic ring, a bridged ring, or a condensed ring. The heterocyclic group can contain 3 to 20 ring atoms, 3 to 10 ring atoms, 3 to 8 ring atoms, 3 to 7 ring atoms, 3 to 6 ring atoms, 4 to 9 ring atoms, 4 to 8 ring atoms, 4 to 7 ring atoms, or 4 to 6 ring atoms. The heteroatom can be any one or more selected from the group consisting of N, O, and S. The heteroatom can be 1 to 3, 1, or 2 heteroatoms selected from the group consisting of N, O, and S.

[0243] Non-limiting examples of heterocyclic groups can include aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, dihydropyridinyl, tetrahydropyridinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, dihydrofuranyl, dihydropyranyl, tetrahydrothiophenyl, morpholinyl, thiomorpholinyl, azepanyl, diazepanyl, oxazepanyl, thioazepanyl, and the like.

[0244] The term "heterocyclyloxy" refers to a functional group in which an oxygen atom is directly attached to the heterocyclic ring.

[0245] The term "heteroaryl" or "heteroarylidene" refers to a monocyclic or bicyclic aromatic moiety containing one or more heteroatoms selected from the group consisting of N, O, and S, with the remaining ring atoms being carbon. A heteroaryl group can contain, for example, 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 or 2 heteroatoms. A heteroaryl group can contain 5 to 10 ring elements, 5 to 7 ring elements, or 5 or 6 ring elements. A heteroaryl group can be a 5- to 6-membered heteroaryl group containing one or two N, O, or S. A heteroaryl group can be a monocyclic group, a bicyclic group, or a tricyclic group. A bicyclic group can be a spirocyclic group, a bridged ring group, and a fused ring group.

[0246] Non-limiting examples of “heteroaryl” may include pyrrolyl, imidazolyl, pyrazolyl, pyridazinyl, furanyl, pyranyl, thienyl, thiophenyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, isothiazol-3-yl, isothiazol-4-yl, isothiazol-5-yl, oxazol-2-yl, oxadiazol-3-yl, oxadiazol-4-yl, oxadiazol-5-yl, oxadi ...1-yl, oxadiazol-1 yl, oxazol-4-yl, oxazol-5-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, 1,2,4-triazol-3-yl, 1,2,4-triazol-5-yl, 1,2,3-triazol-4-yl, 1,2,3-triazol-5-yl, tetrazolyl, pyridin-2-yl, pyridin-3-yl, 2-pyrazin-2-yl, pyrazin-4-yl, pyrazin-5-yl, 2-pyrimidin-2-yl, 4-pyrimidin-2-yl, 5-pyrimidin-2-yl, indolyl, and the like.

[0247] The term "binding functional group" refers to a functional group capable of forming a covalent bond with a functional group contained in a ligand or protein, or a functional group contained in a linker precursor, by addition, substitution, condensation reaction, or the like.

[0248] As used herein, the term "moiety" refers to the portion of a compound that corresponds to the moiety of a parent compound when the parent compound is combined with a compound of Formula 1 or a conjugate of Formula 2. Even when a portion of a whole compound is referred to herein as a compound or active agent, it will be understood from the context that it refers to the "portion" of the compound or active agent.

[0249] In the present invention, the term "precursor" refers to a compound that is a reactant that ultimately provides a desired part through a chemical reaction or the like.

[0250] In the present invention, a "linker precursor" refers to a compound that forms a desired linker structure or a portion thereof through a chemical reaction. For example, various PEG linkers are known, including hydroxyl-PEG linkers, alkynyl-PEG linkers, bromo-PEG linkers, DBCO-PEG linkers, azido-PEG linkers, amino-PEG linkers, and maleimide-PEG linkers. In addition, linker compounds such as aminooxy-PEG linkers, tetrazine-PEG linkers, mesylate-PEG linkers, thiol-PEG linkers, aldehyde-PEG linkers, phosphonate-PEG linkers, hydrazide-PEG linkers, iodine-PEG linkers, and carboxyl-PEG linkers are also widely used in the relevant art. For another example, known linkers having a DBCO (dibenzocyclooctyne) group include amine-reactive DBCO (DBCO-NHS, DBCO-sulfo-NHS ester, DBCO-PEG-NHS ester, DBCO-NHCO-PEG-NHS ester, etc.), carboxyl / carbonyl-reactive DBCO (DBCO-amine, DBCO-PEG-amine, etc.), -SH group-reactive DBCO (DBCO-maleimide, DBCO-PEG-maleimide, etc.), DBCO-PEG-tert-butyl ester, DBCO-alcohol, DBCO-PEG-alcohol, DBCO-PEG-DBCO, bis-DBCO-PEG, and the like. The linker precursors of the present invention include various linkers known in the ligand-conjugate field and are not limited to the linker structures exemplified herein. Useful linker precursors, preparation methods, reaction conditions, etc. are well known in the relevant art.

[0251] The term "dendrimer" refers to an ordered three-dimensional molecular structure having branched units centered around a core.

[0252] The term "1,6-elimination reaction" refers to a reaction in which a covalent bond cleavage occurs at a specific position in the molecular structure of a compound, resulting in the cleavage of a covalent bond at a position 5 atoms away (1,6-position relationship).

[0253] In one embodiment, when β-galactose is cleaved by galactosidase, the compound of Formula 1 having an indole core and a β-galactoside trigger group can release PL- or PL-H through a 1,6-elimination reaction.

[0254] [Reaction Scheme A]

[0255]

[0256] In addition, when the compound of formula 1 contains an optional self-destructive group T, β-galactose and 4-hydroxybenzyl alcohol can be separated to release PL- or PL-H, for example, by a 1,6-elimination reaction as shown in Reaction Scheme B below.

[0257] [Reaction Scheme B]

[0258]

[0259] In addition, when the compound of Formula 1 contains a self-eliminating linker (e.g., -OCO-), β-galactose and CO2 can be separated to release PL - or PL-H, for example, occurs via a 1,6-elimination reaction as shown in Reaction Scheme C below.

[0260] [Reaction Scheme C]

[0261]

[0262] The term "click chemistry reaction" in this application refers to a general term for a molecular assembly reaction that can specifically react with each other using modular reactants under mild conditions such as room temperature and normal pressure. The types of click chemistry reactions and the functional groups involved in click chemistry reactions are generally well known. For example, click chemistry reactions include [3+2] cycloaddition reactions, thiol-ene reactions, Diels-Alder reactions, reverse electron demand Diels-Alder reactions, [4+1] cycloaddition reactions, etc., but are not limited thereto. More specifically, click chemistry reactions include copper (I)-catalyzed azide-alkyne cycloaddition (CuAAC), strain-promoted azide-alkyne cycloaddition (SPAAC), strain-promoted alkyne-nitrone cycloaddition (SPANC), [3+2] cycloaddition of alkenes and azides, reverse demand Diels-Alder reactions of alkenes and tetrazines, photoclick reactions of alkenes and tetrazoles, and Huisgen cycloadditions of azides and alkynes, but are not limited thereto. Click chemistry functional groups include alkynes, cycloalkynes, cyclooctynes ​​and cyclononynes (e.g., cycloalkynes such as bicyclo[6.1.0]-non-4-yn-9-ylmethanol), trans-cyclooctynes, nitrone, nitrile oxide, azide, conjugated dienes, dienophiles and cycloalkynes such as cyclooctyne, cyclononyne, dibenzocyclooctyne (DIBO), BARAC (bisaryl azacyclooctyne ketone), ALO (aryl octyne-free), DIFO (difluorinated cyclooctyne), MOFO (monofluorinated), DIBAC (dibenzoazacyclooctyne) and DIMAC (dimethoxyazacyclooctyne), but are not limited thereto. For example, click chemistry functional groups may include acetylene, trans-cyclooctyne, cyclooctyne, diaryl cyclooctyne, methyl phosphine, norbornene, tetrazine, methylcyclopropene, azetidine, cyanide, azide, dibenzocyclooctyne, etc.

[0263] Medical uses and pharmaceutical compositions

[0264] The ligand-drug conjugates of Formula 1 or Formula 2 of the present invention exhibit excellent stability in various blood environments. In other words, the active agent maintains a stable binding state in the plasma environments of mice, rats, dogs, and humans, thereby minimizing blood toxicity.

[0265] In addition, the ligand-drug conjugates of the compound of Formula 1 or Formula 2 of the present invention can exhibit excellent target cell selectivity and excellent active agent release performance. The compounds and conjugates of the present invention can rapidly dissociate and release the active agent by reacting with enzymes such as galactosidase and glucuronidase under pH conditions of 4 to 5. Therefore, the compounds of the present invention can effectively release the active agent in the target cell intracellular environment (e.g., tumor lysosomes).

[0266] In one aspect of the present invention, a pharmaceutical composition is provided, comprising a compound containing a self-immolative group of Formula 1 or Formula 1-1, or a ligand-drug conjugate of Formula 2 or Formula 2-1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0267] In another aspect of the present invention, an imaging composition or a detection composition is provided, which comprises a compound containing a self-immolative group of Formula 1 or Formula 1-1 or a ligand-drug conjugate of Formula 2 or Formula 2-1 or a pharmaceutically acceptable salt thereof.

[0268] The self-immolative group-containing compound of Formula 1 or Formula 1-1 and the ligand-drug conjugate of Formula 2 or Formula 2-1 of the present invention can be mixed with a solvent and provided as a composition.

[0269] The composition can be prepared as an injectable liquid solution or suspension. In addition, the composition can be prepared in a solid form suitable for injection, as an emulsion or as a polypeptide encapsulated in liposomes. The compound or ligand-drug conjugate of the present invention can be combined with a pharmaceutically acceptable carrier, including any carrier that does not induce the production of antibodies harmful to the subject receiving the carrier. Suitable carriers can generally include slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acid, polyglycolic acid, polymeric amino acids, amino acid copolymers, lipid aggregates, etc. Proteins can be formulated into vaccines in neutral or salt form.

[0270] The composition may contain a diluent such as water, saline, glycerol, ethanol, or the like. Auxiliary substances such as wetting agents or emulsifiers, pH buffers, or the like may be added to the composition. The composition may be administered parenterally by injection, or may be administered by subcutaneous or intramuscular injection. Additional formulations such as suppositories or oral formulations may be provided. Oral compositions may be provided as solutions, suspensions, tablets, pills, capsules, or sustained-release formulations.

[0271] The composition can be administered in a manner compatible with the dosage form. The composition comprises a therapeutically effective amount of a compound of the invention or a ligand-drug conjugate. A therapeutically effective amount refers to a dose in a single dose or multiple dose regimen that is effective for treating or preventing a disease or condition. The administered dose will be determined by the type of active agent contained in the compound of the invention or the ligand-drug conjugate and / or the type of ligand or protein that binds to the receptor. In addition, the administered dose may vary depending on the health and physical condition of the subject being treated, the desired degree of protection, and other relevant factors.

[0272] For example, a therapeutically effective amount of the compound or ligand-drug conjugate of the present invention, or a pharmaceutical composition comprising the same, can be used to treat or prevent proliferative diseases, autoimmune diseases, or infectious diseases.

[0273] For example, the composition can be used to treat cancer or tumors. For example, the composition can be administered to a patient to treat or prevent infection by a pathogen (e.g., a virus, bacteria, fungus, parasite, etc.). These methods include administering to a mammal a therapeutic or prophylactic amount of a compound or conjugate sufficient to treat a disease or disorder or its symptoms, under conditions such that the disease or disorder is prevented or treated.

[0274] In the above-mentioned composition, the compound of the present invention or conjugate can be administered in the form of its pharmaceutically acceptable salt, hydrate or solvate. In one embodiment, it can be administered together with a pharmaceutically acceptable carrier, a pharmaceutically acceptable excipient and / or a pharmaceutically acceptable additive. The pharmaceutically effective amount and the type of pharmaceutically acceptable salt or solvate, excipient and additive can be determined using standard methods (see Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 18th edition, 1990).

[0275] As used herein, the term "pharmaceutically acceptable salt" includes organic salts and inorganic salts. Examples include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate (genticinate), fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1'-methylene-bis(2-hydroxy-3-naphthoate)). Pharmaceutically acceptable salts may include another molecule (e.g., acetate ion, succinate ion and other counterions, etc.), and may also include one or more charged atoms or one or more counterions.

[0276] Exemplary solvates that can be used as pharmaceutically acceptable solvates of the above compounds include, but are not limited to, solvates of water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, or ethanolamine.

[0277] Effects of the Invention

[0278] The compounds or ligand-drug conjugates of the present invention can stably deliver active agents such as drugs, toxins, fluorophores, affinity ligands, diagnostic substances or detection probes to target sites, and can rapidly release the active agents in the specific environment of the target site.

[0279] The compounds or ligand-drug conjugates of the present invention may have excellent stability at blood temperature and neutral conditions, and may rapidly release the active agent under acidic conditions, such as in the tumor microenvironment. BRIEF DESCRIPTION OF THE DRAWINGS

[0280] Figures 1 to 4 The results of measuring the enzymatic cleavage rates of compounds A-15, A-23, A-36, and A-69 using Escherichia coli β-galactosidase or Escherichia coli β-glucuronidase are shown, respectively.

[0281] Figure 5 Shown are the results of chemical stability and plasma stability measurements of Compound A-69.

[0282] Figures 6 to 8 Shown are the results of in vivo activity analysis of ADC-1 to ADC-7 and ADC-11 to ADC-15. DETAILED DESCRIPTION

[0283] The present invention will be described in more detail below by way of examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

[0284] The abbreviations used in this specification are as follows. Abbreviations not listed in the following abbreviation list have meanings commonly used in the field of organic synthesis:

[0285] AcO: acetyl

[0286] AcOH: acetic acid

[0287] EA: ethyl acetate

[0288] MC: dichloromethane

[0289] DMF: dimethylformamide

[0290] EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0291] EDC: N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

[0292] HOBt: 1-Hydroxybenzotriazole hydrate

[0293] ACN: acetonitrile

[0294] THF: Tetrahydrofuran

[0295] DCC: N,N'-dicyclohexylcarbodiimide

[0296] DMAP: 4-dimethylaminopyridine

[0297] NHS: N-hydroxysuccinimide

[0298] DIPEA: diisopropylethylamine

[0299] TEA: triethylamine

[0300] Boc: tert-butyloxycarbonyl

[0301] LAH: lithium aluminum hydride

[0302] TFA: trifluoroacetic acid

[0303] AgOTf: silver trifluoromethanesulfonate

[0304] KO t Bu: Potassium tert-butoxide

[0305] MMAF-OMe: monomethyl auristatin F methyl ester

[0306] PPTS: Pyridinium p-toluenesulfonate

[0307] TBAI: Tetrabutylammonium iodide

[0308] Preparation Example 1: Preparation of Linker P-1

[0309]

[0310] Step 1: Preparation of compounds P-1b and P-1c

[0311] Compound P-1a (tetraethylene glycol, Daejung Chemicals & Metals, CAS No.: 112-60-7, 30 g, 154.46 mmol) was dissolved in MC (300 mL) at 0°C under a nitrogen atmosphere. 4-Methylbenzenesulfonyl chloride (14.7 g, 77.23 mmol) and potassium hydroxide (4.3 g, 77.23 mmol) were then added, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, MC (500 mL) and distilled water (500 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography to yield compounds P-1b and P-1c as colorless oils (P-1b: 4.4 g, 11%; P-1c: 15.1 g, 56%), respectively.

[0312] P-1b: 1 H-NMR (400MHz, CDCl3) δ7.79(d,J=8.4Hz,4H),7.34(d,J=8.0Hz,4H),4.16-4.14(m,4H),3.69-3.66(m,4H),3.57-3.55(m,8H),2.44(s,6H).

[0313] P-1c: 1 H-NMR (400MHz, CDCl3) δ7.80(d,J=8.4Hz,2H),7.34(d,J=8.0Hz,2H),4.18-4.15(m,2H),3.72-3.59(m,14H),2.45(s,3H).

[0314] Step 2: Preparation of compound P-1d

[0315] Compound P-1b (8.78 g, 17.48 mmol) was dissolved in DMF (50 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (3.41 g, 52.44 mmol) was added, and the mixture was stirred at 60 ° C for 16 hours. After the reaction was completed, the mixture was cooled to room temperature, EA (500 mL) and saturated sodium bicarbonate aqueous solution (500 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-1d (4.17 g, 97%) as a colorless liquid.

[0316] 1 H-NMR (400MHz, CDCl3) δ3.69-3.66(m,12H),3.40-3.37(m,4H).

[0317] Step 3: Preparation of linker P-1

[0318] Compound P-1d (4.17 g, 17.06 mmol) was dissolved in EA (32 mL), diethyl ether (32 mL) and 5% aqueous hydrochloric acid solution (64 mL) at 0°C under a nitrogen atmosphere, followed by the addition of triphenylphosphine (4.47 g, 17.06 mmol), and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to remove the organic layer, and then MC (100 mL) was added to the aqueous layer to wash the aqueous layer, and this step was repeated three times. The resulting aqueous layer was concentrated under reduced pressure to obtain linker P-1 (4.12 g, 95%) as a colorless liquid.

[0319] 1 H-NMR (400MHz, CDCl3) δ8.27(brs,2H),3.86-3.83(m,2H),3.72-3.68(m,10H),3.47-3.45(m,2H),3.26-3.24(m,2H).

[0320] Preparation Example 2: Preparation of Linker P-2

[0321]

[0322] Step 1: Preparation of compound P-2a

[0323] Compound P-1c (15.1 g, 43.34 mmol) prepared in Step 1 of Preparation Example 1 was dissolved in DMF (100 mL) at room temperature under a nitrogen atmosphere, followed by the addition of sodium azide (4.2 g, 65.01 mmol), and the mixture was stirred at 60° C. for 16 hours. After the reaction was completed, the mixture was cooled to room temperature, EA (800 mL) and distilled water (800 mL) were added, and the organic layer was extracted five times. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-2a (6.11 g, 64%) as a colorless liquid.

[0324] 1 H-NMR (400MHz, CDCl3) δ3.73-3.66(m,12H),3.62-3.60(m,2H),3.41-3.39(m,2H).

[0325] Step 2: Preparation of compound P-2b

[0326] Compound P-2a (3.82 g, 17.44 mmol) was dissolved in THF (50 mL) at 0°C under nitrogen atmosphere, and then KO was added. t Bu (2.93 g, 26.16 mmol) was added, and the mixture was stirred at 0 ° C for 20 minutes. Then, propargyl bromide (5.2 g, 34.88 mmol) was added, and the mixture was stirred for 16 hours while the temperature was slowly raised from 0 ° C to room temperature. After the reaction was completed, the reaction solution was diluted with EA (200 mL), the solution was filtered with a diatomaceous earth filter, and extracted by adding distilled water (250 mL) to the solution. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue obtained by column chromatography obtained compound P-2b (2.97 g, 66%) in the form of an orange liquid.

[0327] 1 H-NMR (400MHz, CDCl3) δ4.21 (d, J = 2.4Hz, 2H), 3.70-3.67 (m, 14H), 3.40-3.38 (m, 2H), 2.43 (t, J = 2.4Hz, 1H).

[0328] Step 3: Preparation of linker P-2

[0329] Compound P-2b (2.97 g, 11.54 mmol) was dissolved in EA (24 mL), diethyl ether (24 mL) and 5% aqueous hydrochloric acid solution (48 mL) at 0°C under a nitrogen atmosphere, and then triphenylphosphine (3.03 g, 11.54 mmol) was added, and the mixture was stirred for 16 hours while slowly raising the temperature from 0°C to room temperature. After the reaction was completed, the reaction solution was concentrated under reduced pressure to remove the organic solvent, and then MC (100 mL) was added to the remaining aqueous solution to extract and remove the organic layer. The resulting aqueous layer was concentrated under reduced pressure to obtain linker P-2 (2.66 g, 86%) as a yellow liquid.

[0330] 1 H-NMR (400MHz, DMSO-d6) δ7.93(brs,2H),4.14(d,J=2.4Hz,2H),3.62-3.52(m,14H),3.44(t,J=2.4Hz,1H),2.96-2.95(m,2H).

[0331] Preparation Example 3: Preparation of Linker P-3

[0332]

[0333] Step 1: Preparation of compound P-3a

[0334] The compound P-2a (2.27 g, 10.35 mmol) prepared in step 1 of Preparation Example 2 was dissolved in MC (52 mL) under a nitrogen atmosphere at room temperature, and then TEA (4.3 mL, 31.06 mmol) and 4-methylbenzenesulfonyl chloride (3.95 g, 20.71 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, MC (100 mL) and distilled water (100 mL) were added, and the organic layer was extracted three times. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-3a (2.52 g, 65%) as a yellow liquid.

[0335] 1 H-NMR (400MHz, CDCl3) δ7.80(d,J=8.0Hz,2H),7.34(d,J=8.0Hz,2H),4.16(t,J=4.8Hz,2H),3.70-3.59(m,12H),3.37(t,J=4.8Hz,2H),2.45(s,3H).

[0336] Step 2: Preparation of linker P-3

[0337] Compound P-3a (2.52 g, 6.76 mmol) was dissolved in THF (33 mL) at room temperature under a nitrogen atmosphere, and then lithium bromide (1.76 g, 20.29 mmol) was added, and the mixture was refluxed and stirred for 3 hours. After the reaction was completed, the mixture was cooled to room temperature, EA (150 mL) and sodium bicarbonate aqueous solution (150 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-3 (1.8 g, 94%) as a yellow liquid.

[0338] 1 H-NMR (400MHz, CDCl3) δ3.81(t,J=6.4Hz,2H),3.69-3.67(m,10H),3.47(t,J=6.4Hz,2H),3.39(m,2H).

[0339] Preparation Example 4: Preparation of Linker P-4

[0340]

[0341] Step 1: Preparation of compound P-4b

[0342] Compound P-4a (pentaethylene glycol, Merck, CAS No.: 4792-15-8, 10 g, 41.97 mmol) was dissolved in MC (60 mL) at 0°C under a nitrogen atmosphere, followed by the addition of 4-methylbenzenesulfonyl chloride (17.60 g, 92.33 mmol) and lithium hydroxide monohydrate (8.80 g, 209.85 mmol), and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, MC (200 mL) and distilled water (200 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-4b (22.5 g, 98%) as a colorless oil.

[0343] 1 H-NMR (400MHz, CDCl3) δ7.79(d,J=8.0Hz,4H),7.34(d,J=8.0Hz,4H),4.14(t ,J=4.8Hz,4H),3.68(t,J=4.8Hz,4H),3.60(s,4H),3.58(s,8H),2.44(s,6H).

[0344] Step 2: Preparation of compound P-4c

[0345] Compound P-4b (22.5 g, 41.16 mmol) was dissolved in ACN (200 mL) at room temperature under a nitrogen atmosphere, and sodium azide (6.69 g, 102.89 mmol) was added, and the mixture was stirred at 80 ° C for 15 hours. After the reaction was completed, the mixture was cooled to room temperature, filtered using ether (100 mL) to remove the precipitate, and the remaining filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-4c (8.72 g, 73.5%) as a colorless liquid.

[0346] 1 H-NMR (400MHz, CDCl3) δ3.69-3.66 (m, 16H), 3.39 (t, J = 4.8Hz, 4H).

[0347] Step 3: Preparation of linker P-4

[0348] Compound P-4c (8.72 g, 30.25 mmol) was dissolved in EA (48 mL) and diethyl ether (48 mL) at 0° C. under a nitrogen atmosphere, and then 5% aqueous hydrochloric acid solution (96 mL) and triphenylphosphine (8.73 g, 33.28 mmol) were slowly added dropwise. The reaction solution was stirred for 15 hours while slowly raising the temperature from 0° C. to room temperature. After the reaction was completed, the organic layer was removed, and the remaining aqueous layer was washed again with EA (50 mL). The resulting aqueous layer was concentrated under reduced pressure to obtain linker P-4 (7.8 g, 86.3%) in the form of a colorless oil.

[0349] 1 H-NMR (400MHz, CDCl3)8.21(brs,2H),3.90(t,J=4.8Hz,2H),3.74-3.66(m,14H),3.51(t,J=4.8Hz,2H),3.21(m,2H).

[0350] Preparation Example 5: Preparation of Linker P-5

[0351]

[0352] Step 1: Preparation of compound P-5a

[0353] The connector P-1 (3.36 g, 13.19 mmol) was dissolved in 1,4-dioxane (44 mL) at 0 ° C under a nitrogen atmosphere, and then sodium bicarbonate (2.21 g, 26.38 mmol) and di-tert-butyl dicarbonate (Boc anhydride, 3.45 g, 15.83 mmol) dissolved in distilled water (22 mL) were added, and the mixture was stirred at room temperature for 5 hours. After the reaction was completed, EA (350 mL) and distilled water (300 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-5a (3.68 g, 87%) as a colorless liquid.

[0354] 1 H-NMR (400MHz, CDCl3) δ5.02(brs,1H),3.70-3.60(m,10H),3.54(t,J=5.2Hz,2H),3.39(t,J=5.2Hz,2H),3.32-3.31(m,2H),1.44(s,9H).

[0355] Step 2: Preparation of compound P-5b

[0356] THF (70 mL) was added dropwise to sodium hydride (60% dispersion in mineral oil, 924 mg, 23.10 mmol) at 0 ° C under a nitrogen atmosphere, and compound P-5a (3.68 g, 11.55 mmol) dissolved in THF (30 mL) was slowly added to the reaction solution, and the mixture was stirred at 0 ° C for 30 minutes. Iodomethane (7.19 mL, 115.52 mmol) was slowly added to the reaction solution, and the mixture was stirred for 30 minutes, and then stirred for 16 hours while the temperature was slowly raised to room temperature. After the reaction was completed, the mixture was cooled to 0 ° C, and distilled water (350 mL) and EA (350 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained compound P-5b (3.74 g, 97%) in the form of a yellow liquid by column chromatography.

[0357] 1 H-NMR (400MHz, CDCl3) δ3.69-3.62(m,12H),3.41-3.38(m,4H),2.91(s,3H),1.45(s,9H).

[0358] Step 3: Preparation of linker P-5

[0359] Compound P-5b (3.74 g, 11.27 mmol) was dissolved in MC (20 mL) at 0 ° C. under a nitrogen atmosphere, and then a 4M-hydrochloric acid solution (4M HCl in 1,4-dioxocane, 40 mL) was added, and the mixture was stirred at room temperature for 1.5 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain linker P-5 (3.33 g, 99%) in the form of a yellow liquid.

[0360] 1 H-NMR (400MHz, CDCl3) δ9.45(brs,1H),3.91(t,J=5.2Hz,2H),3.72-3.68(m,10H),3.44(t,J=5.2Hz,2H),3.20-3.17(m,2H),2.76(t,J=5.2Hz,3H).

[0361] Preparation Example 6: Preparation of Linker P-6

[0362]

[0363] Compound P-6a (triethylene glycol monomethyl ether, TCI, CAS No.: 112-35-6, 1 g, 6.09 mmol) was dissolved in THF (10 mL) at 0° C. under a nitrogen atmosphere, and then KO was added. t Bu (1 g, 9.13 mmol) was added, and the mixture was stirred at 0 ° C for 20 minutes. Propargyl bromide (1.3 mL, 12.18 mmol) was added to the reaction solution, and the mixture was stirred for 16 hours while slowly raising the temperature from 0 ° C to room temperature. After the reaction was completed, the reaction solution was diluted with EA (200 mL), the solution was filtered through diatomaceous earth, and extracted by adding distilled water (250 mL) to the solution. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain connector P-6 (820 mg, 66%).

[0364] 1 H-NMR (400MHz, CDCl3) δ4.21 (d, J = 2.4Hz, 2H), 3.71-3.63 (m, 10H), 3.56-3.54 (m, 2H), 3.38 (s, 3H), 2.42 (t, J = 2.4Hz, 1H).

[0365] Preparation Example 7: Preparation of Linker P-7

[0366]

[0367] Step 1: Preparation of compound P-7b

[0368] Sodium hydride (60% dispersion in mineral oil, 411 mg, 10.29 mmol) was added to THF (10 mL) at 0°C under a nitrogen atmosphere, and compound P-7a (tetraethylene glycol, Merck, CAS No.: 112-60-7, 6.0 g, 30.89 mmol) was dissolved in THF (20 mL) and slowly added dropwise, and the mixture was stirred for 5 minutes. Propargyl bromide (80% in toluene, 1.146 mL, 10.29 mmol) was added to the reaction solution, and the mixture was stirred for another 2 hours at 0°C. After the reaction was completed, MC (200 mL) and distilled water (100 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound P-7b (1.9 g, 79.8%) in the form of an oil.

[0369] 1 H-NMR (400MHz, CDCl3) δ4.21 (d, J = 2.4Hz, 2H), 3.72-3.67 (m, 14H), 3.61 (t, J = 4.4Hz, 2H), 2.61 (brs, 1H), 2.43 (t, J = 2.4Hz, 1H).

[0370] Step 2: Preparation of compound P-7c

[0371] Compound P-7b (1.0 g, 4.3 mmol) was dissolved in acetone (30 mL), and Jones reagent (Merck, CAS No. 65272-70-0, 3 mL) was slowly added at -5°C. The mixture was stirred at room temperature for 3 hours. After the reaction was completed, the green inorganic salt was removed by filtration, and the remaining filtrate was extracted with MC (100 mL) and distilled water (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound P-7c (630 mg, 59.4%).

[0372] 1 H-NMR (400MHz, DMSO-d6) δ4.20(d,J=2.4Hz,2H),4.12(s,2H),3.73-3.67(m,12H),2.44(t,J=2.4Hz,1H); EI-MS m / z:246[M+H] + .

[0373] Step 3: Preparation of linker P-7

[0374] Compound P-7c (90 mg, 0.365 mmol) was dissolved in MC (5 mL) at 0 ° C under a nitrogen atmosphere, and then NHS (46.2 mg, 0.40 mmol) and DCC (83 mg, 0.40 mmol) were added and the mixture was stirred for 16 hours. After the reaction was completed, EA / n-pentane (1: 1 volume ratio, 20 mL) was added and the resulting precipitate was removed by filtration. The filtrate was concentrated and then EA / n-pentane (1: 1 volume ratio, 20 mL) solution was added again to remove the resulting precipitate. The filtrate was concentrated under reduced pressure and the resulting joint P-7 was used for the next step reaction (100 mg, 80%) without additional purification.

[0375] 1 H-NMR (400MHz, CDCl3) δ4.53 (s, 2H), 4.20 (d, J = 2.4Hz, 2H), 3.81-3.79 (m, 2H), 3.75-3.67 (m, 10H), 2.85 (s, 4H), 2.43 (t, J = 2.4Hz, 1H); EI-MS m / z:366[M+Na] + .

[0376] Preparation Example 8: Preparation of Linker P-8

[0377]

[0378] P-8a (2-[2-(2-aminoethoxy)ethoxy]acetic acid, TCI, CAS No. 134978-97-5, 1.28 g, 7.87 mmol) was dissolved in 1,4-dioxane (30 mL) at 0°C under a nitrogen atmosphere. Sodium bicarbonate (1.32 g, 15.74 mmol) and di-tert-butyl dicarbonate (Boc anhydride, 2.06 g, 9.44 mmol) dissolved in distilled water (15 mL) were then added sequentially. The mixture was stirred for 16 hours while slowly raising the temperature from 0°C to room temperature. After the reaction was complete, EA (200 mL), distilled water (150 mL), and 2N aqueous hydrochloric acid (50 mL) were added, and the organic layer was extracted five times. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain linker P-8 (2.32 g, quantitatively obtained) as a colorless liquid.

[0379] 1 H-NMR (400MHz, CDCl3) δ4.93(brs,1H),4.17(s,2H),3.78-3.76(m,2H),3.68-3.65(m,2H),3.62-3.56(m,2H),3.38-3.32(m,2H),1.45(s,9H).

[0380] Preparation Example 9: Preparation of Auristatin F-OMe

[0381]

[0382] The compound MMAF-OMe (CAS No.: 863971-12-4, 1 g, 1.34 mmol) was dissolved in DMF (7 mL) under a nitrogen atmosphere at room temperature, and then 37% formaldehyde aqueous solution (37 wt.% in H2O, 299 μL, 4.02 mmol) and acetic acid (1.53 mL, 26.81 mmol) were added sequentially, and the mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (NaCNBH3, 168.5 mg, 2.68 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 2.5 hours. After the reaction was completed, saturated sodium bicarbonate aqueous solution was slowly added dropwise until the pH of the reaction solution reached 9, and EA (100 mL) was added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain auristatin F-OMe (892 mg, 88%) in the form of a white solid; EI-MS m / z: 761.0 [M+H] + .

[0383] Preparation Example 10: Preparation of Compound L3-1

[0384]

[0385] Step 1: Preparation of compound L3-1b

[0386] Compound L3-1a (4-hydroxybenzaldehyde, CAS No.: 123-08-0, 300 mg, 2.46 mmol) was dissolved in ACN (10 mL) at 0°C under a nitrogen atmosphere, followed by the addition of acetyl bromide-α-D-galactose (Merck, CAS No.: 3068-32-4, 1.1 g, 2.70 mmol), molecular sieves (100 mg) and silver (I) oxide (1.42 g, 6.14 mmol), and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was diluted with EA (100 mL), filtered through celite, and concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound L3-1b (951 mg, 86%) as a white solid.

[0387] 1 H-NMR (400MHz, CDCl3) δ9.93 (s, 1H), 7.86 (d, J = 8.4Hz, 2H), 7.11 (d, J = 8.4Hz, 2H ),5.47(m,2H),5.15(m,2H),4.17(m,3H),2.19(s,3H),2.07(s,6H),2.02(s,3H).

[0388] Step 2: Preparation of compound L3-1c

[0389] Compound L3-1b (951 mg, 2.1 mmol) was dissolved in THF (20 mL) at 0°C under a nitrogen atmosphere, followed by the addition of sodium borohydride (159 mg, 4.2 mmol), and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, distilled water (100 mL) was added to terminate the reaction, and the organic layer was extracted three times with EA (100 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound L3-1c (695 mg, 73%) as a white solid.

[0390] 1 H-NMR(400MHz, CDCl3)7.31(d,J=8.8Hz,2H),7.00(d,J=8.8Hz,2H),5.46(m,2H) ,5.04(m,2H),4.66(s,2H),4.17(m,3H),2.19(s,3H),2.07(s,6H),2.01(s,3H).

[0391] Step 3: Preparation of compound L3-1

[0392] Compound L3-1c (800mg, 1.76mmol) is dissolved in MC (10mL) at 0°C under a nitrogen atmosphere, then thionyl chloride (191 μL, 2.64mmol) is added, and the mixture is stirred for 2 hours while the temperature is raised from 0°C to room temperature. After the reaction is complete, MC (10mL) is added to dilute the mixture, then concentrated under reduced pressure. The organic layer of the residue obtained by concentrated under reduced pressure is extracted twice with EA (50mL) and distilled water (50mL). The gained organic layer is dried over anhydrous magnesium sulfate, filtered, then concentrated under reduced pressure to obtain compound L3-1 (831mg, 99%) in the form of a colorless viscous oil.

[0393] 1 H-NMR (400MHz, CDCl3)7.32(d,J=8.8Hz,2H),6.98(d,J=8.8Hz,2H),5.51-5.45(m,2H),5.11(dd,J=10.4, 3.6Hz,1H),5.04(d,J=7.6Hz,1H),4.56(s,2H),4.25-4.14(m,3H),2.18(s,3H),2.06(s,6H),2.01(s,3H).

[0394] Preparation Example 11: Preparation of Compound PL-1

[0395]

[0396] Step 1: Preparation of compound PL-1b

[0397] A solution of compound PL-1a (benzyloxycarbonyl-L-valine, Z-Val-OH, Merck, CAS No.: 1149-26-4, 5 g, 19.9 mmol) dissolved in THF (20 mL) was slowly added to a reaction solution at 0°C under a nitrogen atmosphere. The reaction solution was obtained by adding THF (50 mL) to 60% sodium hydride (60% dispersion in mineral oil, 3.4 g, 85.56 mmol), and the mixture was stirred at 0°C for 20 minutes. Then, iodomethane (12.38 mL, 198.98 mmol) was slowly added, and the mixture was stirred at room temperature for 20 hours. The reaction solution was cooled to 0°C, distilled water (150 mL) was added to terminate the reaction, EA (200 mL) and 2N-hydrochloric acid aqueous solution (50 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound PL-1b (3.7 g, 70%).

[0398] 1 H-NMR (400MHz, DMSO-d6) δ7.36-7.30(m,5H),5.11(s,2H),4.22-412(m,1H),2.84-81(m,3H),2.11(m,1H),0.95(m,3H),0.82(m,3H).

[0399] Step 2: Preparation of compound PL-1d

[0400] Compound PL-1b (600 mg, 2.26 mmol) and compound PL-1c (N-methylaniline, Merck, CAS No.: 100-61-8, 361 mg, 3.39 mmol) were dissolved in MC (12 mL) at 0 ° C under a nitrogen atmosphere, and then DCC (700 mg, 3.39 mmol), DMAP (55.2 mg, 0.46 mmol) and DIPEA (0.78 mL, 4.52 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 20 minutes. The reaction mixture was stirred for another 16 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and extracted by adding 2N-hydrochloric acid aqueous solution (100 mL) to the filtered solution. The obtained organic layer was extracted once with 2N-sodium hydroxide aqueous solution (100 mL) and then washed by adding saturated sodium chloride aqueous solution (100 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound PL-1d (429 mg, 53%) as a colorless oil.

[0401] 1 H-NMR (400MHz, CDCl3) δ7.41-7.24(m,6H),7.09-7.02(m,4H),5.01-4.96(m,2H),4.40-4.28( m,1H),3.25(s,3H),2.91-2.87(m,3H),2.33(m,1H),0.90(d,J=6.8Hz,3H),0.74-0.65(m,3H).

[0402] Step 3: Preparation of compound PL-1

[0403] Compound PL-1d (429 mg, 1.21 mmol) was dissolved in methanol (50 mL) at room temperature under a nitrogen atmosphere, and then 5% palladium on carbon (5% Pd / C, 150 mg) was added, and the mixture was stirred under a hydrogen atmosphere for 2 hours. After completion of the reaction, the reaction solution was diluted with EA (100 mL), filtered through a celite filter, and the solution was concentrated under reduced pressure to obtain compound PL-1 (264.3 mg, 99%) as a colorless oil.

[0404] 1 H-NMR (400MHz, CDCl3) δ7.45-4.41(m,2H),7.37-7.34(m,1H),7.02-7.18(m,2H),3.31(s,3H), 2.85(d,J=6.4Hz,1H),2.31(s,3H),1.71(m,1H),0.85(d,J=6.8Hz,3H),0.80(d,J=6.8Hz,3H).

[0405] Preparation Example 12: Preparation of Compound PL-2

[0406]

[0407] Compound PL-2a (SN-38, 7-ethyl-10-hydroxycamptothecin, ASTATECH, CAS No.: 86639-52-3, 30.0 mg, 0.076 mmol) was dissolved in MC (6.0 mL) under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (23.1 mg, 0.114 mmol) and DIPEA (13.3 μL, 0.076 mmol) and pyridine (18.4 μL, 0.229 mmol) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and then lyophilized to obtain compound PL-2 (7.5 mg, 17%) as a white solid; EI-MS m / z: 558 [M+H] + .

[0408] Preparation Example 13: Preparation of Compound PL-3

[0409]

[0410] Compound PL-3a (abiraterone, TCI, CAS No. 154229-19-3, 30.0 mg, 0.086 mmol) was dissolved in THF (3.0 mL) under a nitrogen atmosphere. Carbonyldiimidazole (CDI, Alfa Aesar, CAS No. 530-62-1, 17.2 mg, 0.105 mmol) was then added, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, EA (10 mL) and distilled water (10 mL) were added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was purified by column chromatography to yield compound PL-3 (16.0 mg, 42%) as a white solid.

[0411] 1H-NMR (400MHz, CDCl3) δ8.62 (s, 1H), 8.46 (d, J = 4.8Hz, 1H), 8.13 (s, 1H), 8.13 (s, 1H), 7.65 (d,J=8.0Hz,1H),7.42(s,1H),7.22(dd,J=8.0Hz,J=4.8Hz,1H),7.06(s,1H),6.00(s,1H), 5.49(d,J=4.8Hz,1H),4.85-4.83(m,1H),2.54-2.52(m,2H),2.29-2.25(m,1H),2.08-2.04 (m,4H),1.97-1.81(m,1H),1.80-1.48(m,9H),1.26-1.22(m,1H),1.13(s,3H),1.06(s,3H).

[0412] Preparation Example 14: Preparation of Core C-1

[0413]

[0414] Step 1: Preparation of compound C-1b

[0415] Compound C-1a (2-benzyloxybenzaldehyde, CAS No. 5896-17-3, 5.48 g, 25.8 mmol) was dissolved in methanol (55 mL) at -30°C under a nitrogen atmosphere, followed by the slow addition of ethyl azidoacetate (CAS No. 637-81-0, TCI, 28.2 mL, 258.20 mmol) and sodium methoxide (25 wt% in MeOH, 44.6 mL, 206.4 mmol). The mixture was stirred at -30°C for 3 hours, then the reaction was terminated by the slow dropwise addition of distilled water (100 mL). The organic layer was extracted with EA (100 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to yield compound C-1b (7.74 g, 97%). Compound C-1b was used in the next step without further purification.

[0416] Step 2: Preparation of compound C-1c

[0417] Compound C-1b (7.74 g, 25.02 mmol) was dissolved in xylene (200 mL) at room temperature under a nitrogen atmosphere, and the mixture was stirred at 150°C for 3 hours. After the reaction was complete, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by column chromatography to afford compound C-1c (510.8 mg, 22.7%).

[0418] 1H-NMR(400MHz, CDCl3)δ8.82(brs,1H),7.51-7.49(m,2H),7.42-7.33(m,4H),7.22 (t,J=8Hz,1H),7.02(d,J=8Hz,1H),6.58(d,J=8Hz,1H),5.22(s,2H),3.92(s,3H).

[0419] Step 3: Preparation of compound C-1d

[0420] Compound C-1c (1.14 g, 4.05 mmol) was dissolved in DMF (35 mL) at room temperature under a nitrogen atmosphere, followed by the addition of potassium carbonate (1.68 g, 12.15 mmol) and iodomethane (0.65 mL, 10.53 mmol), and the mixture was stirred for 12 hours. After the reaction was complete, EA (100 mL) and distilled water (100 mL) were added, and the organic layer was extracted. Distilled water (300 mL) was added and the resulting organic layer was extracted again. The organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound C-1d (912.7 mg, 76%).

[0421] 1 H-NMR (400MHz, CDCl3) δ7.50-7.47(m,3H),7.42-7.33(m,3H),7.25(m,1H),6. 99(d,J=8Hz,1H),6.57(d,J=8Hz,1H),5.22(s,2H),4.06(s,3H),3.89(s,3H).

[0422] Step 4: Preparation of compound C-1e

[0423] C-1d (800 mg, 2.70 mmol) was dissolved in MC (40 mL) at -50°C under a nitrogen atmosphere, followed by the slow addition of dichloromethyl methyl ether (735 μL, 8.10 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 8.1 mL, 8.1 mmol). The mixture was stirred for 2 hours. After the reaction was complete, cooled distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound C-1e (392 mg, 44.8%).

[0424] 1H-NMR (400MHz, CDCl3) δ10.10 (s, 1H), 7.81 (d, J = 8.4Hz, 1H), 7.44 (s, 1H), 7.49-7.47 (m ,2H),7.44-7.36(m,3H),6.69(d,J=8.4Hz,1H),5.39(s,2H),4.36(s,3H),3.90(s,3H).

[0425] Step 5: Preparation of Core C-1

[0426] Compound C-1e (393.2 mg, 1.22 mmol) was dissolved in MC (20 mL) at -50 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 6 mL, 6.08 mmol) was slowly added, and the mixture was stirred for 1.5 hours while slowly warming to -30 ° C. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (100 mL) was added to the mixture, and the organic layer was extracted four times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain core C-1 (239.7 mg, 84%) in the form of a light orange solid.

[0427] 1 H-NMR (400MHz, DMSO-d6) δ9.99(s,1H),7.79(m,1H),7.43(s,1H),6.55(m,1H),4.22(s,3H),3.84(s,3H).

[0428] Preparation Example 15: Preparation of Core C-2

[0429]

[0430] Step 1: Preparation of compound C-2b

[0431] Compound C-2a (2-nitro-m-cresol, TCI, CAS No.: 4920-77-8, 20 g, 130.59 mmol) was dissolved in ACN (250 mL) at 0° C. under a nitrogen atmosphere, followed by the addition of benzyl chloride (15.78 mL, 137.12 mmol) and potassium carbonate (22.56 g, 163.24 mmol), and the mixture was stirred at 80° C. for 16 hours. After the reaction was complete, EA (200 mL) and distilled water (300 mL) were added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to give compound C-2b (31.76 g, 99%) as a light yellow oil.

[0432] 1H-NMR (400MHz, CDCl3) δ7.37-7.31(m,5H),7.24(m,1H),6.89-6.83(m,2H),5.15(s,2H),2.31(s,3H).

[0433] Step 2: Preparation of compound C-2c

[0434] Will KO t Bu (15.35 g, 136.82 mmol) was dissolved in THF (100 mL) and diethyl ether (300 mL) at room temperature under a nitrogen atmosphere. Diethyl oxalate (Merck, CAS No.: 95-92-1, 19.46 mL, 143.34 mmol) was then added, and the mixture was stirred for 15 minutes. Compound C-2c (31.7 g, 31.7 mmol) was dissolved in THF (50 mL) and slowly added to the reaction solution. The mixture was stirred at room temperature for 19 hours and then at 80°C for 3 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and diethyl ether (200 mL) was added dropwise. The resulting precipitate was filtered, washed with diethyl ether (100 mL), and dried to obtain compound C-2c (41.5 g, 83%) as a light orange solid.

[0435] 1 H-NMR(400MHz,DMSO-d6)δ8.66(d,J=8Hz,1H),7.40-7.30(m,5H),7.04(t,J=8Hz,1H),6. 50(d,J=8Hz,1H),5.15(s,1H),5.10(s,2H),3.99(q,J=7.2Hz,2H),1.17(t,J=7.2Hz,3H).

[0436] Step 3: Preparation of compound C-2d

[0437] Iron powder (57.1 g, 1.02 mol) is slowly added to acetic acid (200 mL) at room temperature under a nitrogen atmosphere, and compound C-2c (39 g, 102.24 mmol) dissolved in acetic acid (150 mL) is added under stirring, and the mixture is stirred at 80 ° C for 6 hours. After the reaction is completed, the temperature of the reaction solution is cooled to 45 ° C, EA (500 mL) is added dropwise, the mixture is stirred for another 30 minutes, and the filtrate is obtained by diatomaceous earth filtration, and the filtrate is concentrated under reduced pressure. EA (300 mL) is added to the concentrate thus obtained, and the resulting precipitate is filtered again. The filtrate is concentrated under reduced pressure. The resulting residue obtains compound C-2d (22.18 g, 73%) in the form of a light yellow solid through column chromatography.

[0438] 1H-NMR (400MHz, CD3OD-d4)7.57-7.55(m,2H),7.43-7.39(m,2H),7.36-7.32(m,1H),7.23(d,J=8Hz,1H),7.15 (s,1H),6.98(t,J=8Hz,1H),6.83(d,J=7.6Hz,1H),5.29(s,2H),4.39(q,J=7.2Hz,2H),1.42(t,J=7.2Hz,3H).

[0439] Step 4: Preparation of compound C-2e

[0440] Compound C-2d (6 g, 20.31 mmol) was dissolved in DMF (30 mL) at 0 ° C under a nitrogen atmosphere, and potassium carbonate (4.21 g, 30.46 mmol) and iodomethane (1.64 mL, 26.40 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 1 hour. The reaction temperature was raised to room temperature, and the mixture was stirred for 12 hours, and then the mixture was stirred at 60 ° C for another 3 hours to complete the reaction. The temperature of the reaction solution was cooled to room temperature, and EA (200 mL) and distilled water (400 mL) were added to extract the organic layer. Distilled water (300 mL) was added to extract the obtained organic layer again, and the extracted organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained C-2e was used for the next reaction without additional purification (6.28 g, 99%).

[0441] 1 H-NMR (400MHz, CDCl3) δ7.49-7.47(m,2H),7.42-7.39(m,2H),7.37-7.33(m,1H),7.25-7.23(m,2H),6.99(t ,J=8Hz,1H),6.77(d,J=7.6Hz,1H),5.18(s,2H),4.37(s,3H),4.35(q,J=7.2Hz,2H),1.40(t,J=7.2Hz,3H).

[0442] Step 5: Preparation of compound C-2f

[0443] Compound C-2e (500 mg, 1.61 mmol) was dissolved in MC (20 mL) at -30°C under a nitrogen atmosphere, followed by the slow addition of dichloromethyl methyl ether (Merck, CAS No.: 4885-02-3, 438 μL, 4.83 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 4.85 mL, 4.83 mmol), and the mixture was stirred for 1 hour. After the reaction was complete, cooled distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound C-2f (495 mg, 94%).

[0444] 1 H-NMR (400MHz, CD3OD-d4) δ9.96 (s, 1H), 7.91 (s, 1H), 7.67 (d, J = 8Hz, 1H), 7.57-7.55 (m, 2H), 7.47-7.37(m,3H),7.08(d,J=8Hz,1H),5.37(s,2H),4.39-4.36(m,5H),1.42(t,J=7.2Hz,3H).

[0445] Step 6: Preparation of Core C-2

[0446] Compound C-2f (300 mg, 0.89 mmol) was dissolved in ethanol (8 mL) and THF (4 mL) at room temperature, and then lithium hydroxide monohydrate (75 mg, 1.78 mmol) dissolved in distilled water (3 mL) was slowly added dropwise, and the mixture was stirred at 50° C. for 30 minutes. The reaction solution was cooled, and then 1N-hydrochloric acid aqueous solution (10 mL) and distilled water (50 mL) and EA (100 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain core C-2, which was used in the next reaction without additional purification (275 mg, 99%).

[0447] 1 H-NMR (400MHz, Aceton-d4) δ10.06 (s, 1H), 8.03 (s, 1H), 7.71 (d, J = 8Hz, 1H), 7.6 5-7.63(m,2H),7.49-7.38(m,3H),7.15(d,J=8Hz,1H),5.45(s,2H),4.41(s,3H).

[0448] Preparation Example 16: Preparation of Core C-3

[0449]

[0450] Step 1: Preparation of compound C-3b

[0451] Compound C-3a (benzo[b]thiophene-4-ol, Ambeed, CAS No.: 3610-02-4, 500 mg, 3.35 mmol) was dissolved in 10% potassium hydroxide aqueous solution (410 mg dissolved in 4 mL H2O) at 0°C under a nitrogen atmosphere, and then glycolic acid monohydrate (300 mg) was added and the mixture was stirred for 7 hours. After the reaction was completed, the mixture was diluted by adding distilled water (2 mL), and 2N-hydrochloric acid aqueous solution was slowly added dropwise to adjust the pH of the reaction solution to 7. The organic layer was extracted with ether (10 mL) and removed. 2N-hydrochloric acid aqueous solution was slowly added dropwise to the aqueous layer, the pH was adjusted to 2, and extracted three times with ether (100 mL). The organic layer was collected, dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain compound C-3b (460 mg, 61.6%).

[0452] 1 H-NMR (400MHz, CDCl3) δ7.51(d,J=5.6Hz,1H),7.36(d,J=5.6Hz,1H),7.24(d,J=8Hz,1H),6.72(d,J=8Hz,1H),5.41(s,1H).

[0453] Step 2: Preparation of Core C-3

[0454] Compound C-3b (450 mg, 2.0 mmol) was dissolved in ethanol (0.8 mL) at room temperature under a nitrogen atmosphere, and then ferric sulfate hydrate (923 mg, 2.3 mmol) and 0.4N-sulfuric acid aqueous solution (3.8 mL) were added in sequence, and the mixture was stirred at 60°C for 1 hour. After the reaction was completed, the reaction temperature was lowered to room temperature, and the reaction solution was filtered with EA (100 mL). Distilled water (100 mL) was added to the filtrate to extract the organic layer, which was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain core C-3 (210 mg, 58%).

[0455] 1 H-NMR (400MHz, CDCl3) δ10.01 (s, 1H), 7.74 (d, J = 8Hz, 1H), 7.60 (d, J = 5.6Hz, 1H), 7.52 (d, J = 5.6Hz, 1H), 6.88 (d, J = 8Hz, 1H).

[0456] Preparation Example 17: Preparation of Core C-4

[0457]

[0458] Step 1: Preparation of compound C-4b

[0459] Compound C-4a (benzo[b]thiophene-4-ol, Ambeed, CAS No.: 3610-02-4 (1.14 g, 7.63 mmol) was dissolved in ACN (50 mL) at 0°C under a nitrogen atmosphere, and then potassium carbonate (2.63 g, 19.07 mmol) and benzyl bromide (1 mL, 8.39 mmol) were slowly added, and the mixture was stirred at 0°C for 30 minutes. The reaction temperature was raised to room temperature, and the mixture was stirred for another 16 hours. After the reaction was completed, EA (350 mL) and distilled water (350 mL) were added, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound C-4b (1.87 g, 99%) as a light orange oil.

[0460] 1 H-NMR (400MHz, CDCl3) δ7.57 (d, J = 5.6Hz, 1H), 7.50-7.47 (m, 3H), 7.42-7.38 ( m,2H),7.35-7.32(m,2H),7.27-7.23(m,1H),6.81(d,J=8Hz,1H),5.22(s,2H).

[0461] Step 2: Preparation of compound C-4c

[0462] Compound C-4b (1.87 g, 7.8 mmol) was dissolved in THF (60 mL) at room temperature under a nitrogen atmosphere. The solution was cooled to -78 ° C, then n-BuLi solution (2.5 M n-BuLi in hexane, 1.42 mL, 3.56 mmol) was slowly added dropwise, and the mixture was stirred at the same temperature for 30 minutes. DMF (0.78 mL) was slowly added to the mixture, and the mixture was stirred at the same temperature for another 30 minutes. Distilled water (300 mL) was added to the mixture at the same temperature to terminate the reaction, and the organic layer was then extracted with EA (300 mL). The gained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained compound C-4c (1.36 g, 65%) in the form of a light yellow oil through column chromatography.

[0463] 1 H-NMR (400MHz, CDCl3) δ10.06 (s, 1H), 8.25 (s, 1H), 7.50-7.37 (m, 7H), 6.85 (d, J = 7.6Hz, 1H), 5.24 (s, 2H).

[0464] Step 3: Preparation of Core C-4

[0465] Compound C-4c (300 mg, 1.11 mmol) was dissolved in MC (20 mL) at -78°C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 2.2 mL, 2.20 mmol) was slowly added, and the mixture was stirred for 30 minutes. After the reaction was completed, EA (10 mL) and distilled water (10 mL) were slowly added dropwise at -50°C to terminate the reaction. EA (100 mL) and distilled water (100 mL) were added at room temperature to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain core C-4 (174 mg, 87.4%) as a light yellow-brown solid.

[0466] 1 H-NMR (400MHz, DMSO-d6) δ10.62(brs,1H),8.41(s,1H),7.46(d,J=8Hz,1H),7.39(t,J=8Hz,1H),6.82(d,J=8Hz,1H).

[0467] Preparation Example 18: Preparation of Core C-5

[0468]

[0469] Step 1: Preparation of compound C-5a

[0470] Compound C-4c (1.36 g, 5.09 mmol) prepared in step 2 of Preparation Example 17 was dissolved in methanol (65 mL) at 0 ° C under a nitrogen atmosphere, and then potassium hydroxide (713 mg, 12.72 mmol) dissolved in methanol (15 mL) and iodine (1.67 g, 6.61 mmol) dissolved in methanol (15 mL) were slowly added in sequence, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the organic layer was extracted with EA (500 mL) and distilled water (500 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound C-5a (1.43 g, 94%) as an ivory solid.

[0471] 1 H-NMR (400MHz, CDCl3) δ8.28 (s, 1H), 7.49-7.47 (m, 2H), 7.44-7.34 (m, 5H), 6.82 (d, J = 7.6Hz, 1H), 5.21 (s, 2H), 3.93 (s, 3H).

[0472] Step 2: Preparation of compound C-5b

[0473] Compound C-5a (240 mg, 0.80 mmol) was dissolved in MC (15 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (218 μL, 2.41 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 2.41 mL, 2.41 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 1 hour. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound C-5b (206.7 mg, 79%).

[0474] 1 H-NMR (400MHz, CDCl3) δ10.07(s,1H),8.34(s,1H),7.93(d,J=8Hz,1H),7.50-7.39(m,5H),7.00(d,J=8Hz,1H),5.34(s,2H),3.95(s,3H).

[0475] Step 3: Preparation of Core C-5

[0476] Compound C-5b (82.5 mg, 0.25 mmol) was dissolved in MC (5 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 2.78 mL, 2.78 mmol) was slowly added, and the mixture was stirred for 5 hours while slowly raising the temperature to -30 ° C. After the reaction was completed, distilled water (50 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (50 mL) was added to the mixture, and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain core C-5 (32.8 mg, 55%) in the form of an ivory solid.

[0477] 1 H-NMR (400MHz, DMSO-d6) δ10.01(s,1H),8.23(s,1H),8.12(d,J=8Hz,1H),7.02(d,J=8Hz,1H),3.90(s,3H).

[0478] Preparation Example 19: Preparation of Core C-6

[0479]

[0480] Step 1: Preparation of compound C-6b

[0481] Compound C-6a (3-fluorophenol, TCI, CAS No.: 372-20-3, 10 g, 89.2 mmol) was dissolved in anhydrous ACN (200 mL) at 0 ° C under a nitrogen atmosphere, and then benzyl bromide (12.7 mL, 107 mmol) and potassium carbonate (18.5 g, 7.98 mmol) were added, and the mixture was stirred at room temperature for 20 hours. After the reaction was completed, the organic layer was extracted twice with EA (350 mL) and distilled water (350 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound C-6b (15 g, 83%) as a white solid.

[0482] 1 H-NMR (400MHz, CDCl3) δ7.43-7.37(m,4H),7.35-7.32(m,1H),7.25-7.19(m,1H),6.75(dd,J=8.4,2.4Hz,1H),6.71-6.64(m,2H),5.04(s,2H).

[0483] Step 2: Preparation of compound C-6c

[0484] Compound C-6b (15 g, 74.1 mmol) was added to THF (495 mL) at room temperature under a nitrogen atmosphere, the mixture was cooled to -78 ° C, and then n-butyllithium solution (2.5 M n-BuLi in hexane, 35.6 mL, 89.0 mmol) was slowly added dropwise. The mixture was stirred for 30 minutes while maintaining at -78 ° C, then DMF (17.2 mL) was slowly added dropwise, and the mixture was stirred for another 30 minutes. After 30 minutes, the temperature was slowly raised, and the mixture was then stirred at 0 ° C for 1 hour. After the reaction was completed, the mixture was cooled to -20 ° C again, and distilled water (300 mL) was added dropwise to terminate the reaction. The reaction solution was extracted twice with EA (500 mL), and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained compound C-6c (14.5 g, 85%) as a white solid by column chromatography.

[0485] 1 H-NMR (400MHz, CDCl3) δ10.51 (s, 1H), 7.44-7.40 (m, 6H), 6.83 (d, J = 8.4Hz, 1H), 6.74 (t, J = 8.4Hz, 1H), 5.20 (s, 2H).

[0486] Step 3: Preparation of compound C-6d

[0487] Compound C-6c (14.5 g, 62.9 mmol) was dissolved in DMF (150 mL) at room temperature under a nitrogen atmosphere, followed by addition of methyl thioglycolate (Merck, CAS No.: 2365-48-2, 8.44 mL, 94.4 mmol) and triethylamine (22.1 mL, 157.4 mmol), and the mixture was stirred at 100 ° C for 16 hours. After the reaction was completed, the temperature of the reaction solution was cooled to room temperature, and the organic layer was extracted twice with EA (400 mL) and distilled water (400 mL). The obtained organic layer was washed with sodium chloride aqueous solution. The washed organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained was obtained by column chromatography as a white solid compound C-6d (7.2 g, 38%).

[0488] 1 H-NMR (400MHz, CDCl3) δ8.28 (s, 1H), 7.49-7.47 (m, 2H), 7.45-7.29 (m, 5H), 6.81 (d, J = 7.6Hz, 1H), 5.21 (s, 2H), 3.92 (s, 3H).

[0489] Step 4: Preparation of compound C-6e

[0490] Compound C-6d (1 g, 3.35 mmol) was dissolved in MC (65 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (Merck, CAS No.: 4885-02-3, 910 μL, 10.05 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 10.05 mL, 10.05 mmol) were slowly added in sequence, and the mixture was stirred for 3 hours while maintaining the temperature. After the reaction was completed, distilled water (250 mL) was slowly added dropwise to terminate the reaction, and MC (250 mL) was added to extract the organic layer twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound C-6e (733 mg, 67%).

[0491] 1 H-NMR (400MHz, CDCl3) δ10.07(s,1H),8.34(s,1H),7.93(d,J=8Hz,1H),7.50-7.39(m,5H),7.00(d,J=8Hz,1H),5.34(s,2H),3.95(s,3H).

[0492] Step 5: Preparation of Core C-6

[0493] Compound C-6e (1.37 g, 4.19 mmol) was dissolved in ethanol (45 mL) and THF (45 mL) at room temperature under a nitrogen atmosphere, and then lithium hydroxide monohydrate (352.2 mg, 8.39 mmol) dissolved in distilled water (17 mL) was slowly added dropwise, and the mixture was stirred at room temperature for 1 hour. The mixture was stirred at 60°C for another 3 hours. After the reaction was completed, EA (300 mL) and 2N-hydrochloric acid aqueous solution (250 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain core C-6 (1.3 g, 99%).

[0494] 1 H-NMR (400MHz, DMSO-d6) δ10.08(s,1H),8.26(d,J=8.4Hz,1H),8.09(s,1H),7.56(m,2H),7.46-7.35(m,4H),5.47(s,2H).

[0495] Preparation Example 20: Preparation of Core C-7

[0496]

[0497] Step 1: Preparation of compound C-7a

[0498] Compound C-4c (2.1 g, 7.82 mmol) prepared in step 2 of Preparation Example 17 was dissolved in MC (40 mL) under a nitrogen atmosphere at room temperature, and then (triphenylphosphine) ethyl acetate (5.45 g, 15.6 mmol) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the organic layer was extracted twice with MC (100 mL) and distilled water (100 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound C-7a (2.5 g, 95%) as a white solid.

[0499] 1 H-NMR (400MHz, CDCl3) δ7.85 (d, J = 15.6Hz, 1H), 7.69 (s, 1H), 7.48-7.46 (m, 2H), 7.43-7.35 (m, 4H), 7.31-7.27 ( m,1H),6.79(d,J=7.6Hz,1H),6.25(d,J=15.6Hz,1H),5.21(s,2H),4.26(q,J=7.2Hz,2H),1.33(t,J=7.2Hz,3H).

[0500] Step 2: Preparation of compound C-7b

[0501] Compound C-7a (4.7 g, 13.3 mmol) was dissolved in THF (150 mL) and methanol (500 mL) at room temperature under a nitrogen atmosphere. 5% palladium on carbon (5% Pd / C, 5.67 g) was added, and the mixture was reacted under a hydrogen atmosphere for 1 hour. After completion of the reaction, the solution was filtered through celite and concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound C-7b (4.3 g, 91%) as a white solid.

[0502] 1 H-NMR (400MHz, CDCl3) δ7.48-7.41(m,1H),7.42-7.34(m,4H),7.26-7.25(m,1H),7.18(t,J=8Hz,1H),6.77(d,J=7 .6Hz,1H),5.19(s,2H),4.15(q,J=7.2Hz,2H),3.22(t,J=8.0Hz,2H),2.74(t,J=8.0Hz,2H),1.25(t,J=7.2Hz,3H).

[0503] Step 3: Preparation of compound C-7c

[0504] Compound C-7b (485 mg, 1.42 mmol) was dissolved in MC (30 mL) at -78°C under a nitrogen atmosphere, followed by the slow addition of dichloromethyl methyl ether (Merck, CAS No. 4885-02-3, 400 μL, 4.27 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 4.3 mL, 4.27 mmol), and the mixture was stirred for 2 hours. After the reaction was completed, distilled water (250 mL) was slowly added dropwise to terminate the reaction, and MC (250 mL) was added to extract the organic layer twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound C-7c (290 mg, 55%).

[0505] 1 H-NMR (400MHz, CDCl3) δ10.03(s,1H),7.74(d,J=8.4Hz,1H),7.49-7.35(m,6H),6.94(d,J=8.4Hz,1H), 5.30(s,2H),4.15(q,J=7.2Hz,2H),3.27(t,J=7.6Hz,2H),2.78(t,J=7.6Hz,2H),1.24(t,J=7.2Hz,3H).

[0506] Step 4: Preparation of Core C-7

[0507] Compound C-7c (290 mg, 0.79 mmol) was dissolved in ethanol (5 mL) and THF (5 mL) at room temperature under a nitrogen atmosphere. Lithium hydroxide monohydrate (66 mg, 1.57 mmol) dissolved in distilled water (2.5 mL) was then slowly added dropwise, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, EA (300 mL) and 2N aqueous hydrochloric acid (250 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to yield Core C-7 (278 mg, 99%).

[0508] 1 H-NMR (400MHz, CDCl3) δ10.03 (s, 1H), 7.75 (d, J = 8.4Hz, 1H), 7.49-7.35 (m, 6H), 6.94 (d, J=8.4Hz,1H),5.31(s,2H),3.28(t,J=7.6Hz,2H),2.85(t,J=7.6Hz,2H);EI-MSm / z:341(M + ).

[0509] Preparation Example 21: Preparation of Core C-8

[0510]

[0511] Compound C-7c (279 mg, 0.81 mmol) was dissolved in MC (16 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 2.44 mL, 2.44 mmol) was slowly added and the mixture was stirred for 1.5 hours. After the reaction was completed, the temperature was raised to -50 ° C and distilled water (15 mL) was added dropwise to terminate the reaction. The temperature was raised to 0 ° C and 2N-sodium hydroxide aqueous solution (5 mL) was added dropwise. The organic layer was extracted twice from the mixture using MC (30 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain core C-8 (150 mg, 66%) in the form of a white solid.

[0512] 1 H-NMR (400MHz, CDCl3) δ10.02(s,1H),7.67(d,J=7.6Hz,1H),6.84(d,J=8Hz,1H),5.94(s,1H ), 4.16 (q, J = 6.8Hz, 2H), 3.29 (t, J = 7.6Hz, 2H), 2.79 (t, J = 7.6Hz, 2H), 1.26 (t, J = 7.2Hz, 3H).

[0513] Preparation Example 22: Preparation of Core C-9

[0514]

[0515] Compound C-9a (2-iodoresorcinol, CAS No. 41046-67-7, TCI, 1 g, 4.23 mmol) was dissolved in ACN (150 mL) at 0°C under a nitrogen atmosphere. Methyl 5-hexenoate (CAS No. 77758-51-1, Thermoscientific, 600 μL), PdCl2(TPP)2 (100 mg), CuI (40 mg), and TEA (5.9 mL, 42.3 mmol) were then added sequentially, and the mixture was stirred at room temperature for 1 hour. The mixture was stirred at 60°C for an additional 16 hours, after which EA (100 mL) and 2N aqueous hydrochloric acid (100 mL) were added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to yield Core C-9 (340 mg, 34.2%).

[0516] 1 H-NMR(400MHz, CDCl3)δ7.08-7.01(m,2H),6.59(m,1H),6.47(s,1H),5.15(s ,1H),3.67(s,3H),2.82(t,J=7.2Hz,2H),2.41(t,J=7.2Hz,2H),2.08(m,2H).

[0517] Preparation Example 23: Preparation of Core C-10

[0518]

[0519] Step 1: Preparation of compound C-10a

[0520] Core C-9 (690 mg, 2.94 mmol) was dissolved in ACN (20 mL) at 0°C under a nitrogen atmosphere, followed by the slow addition of potassium carbonate (1.01 g, 7.36 mmol) and benzyl bromide (420 μL, 3.53 mmol). The mixture was stirred at 0°C for 30 minutes and then at room temperature for 16 hours. After the reaction was complete, EA (300 mL) and distilled water (300 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to afford compound C-10a (894 mg, 93%) as a yellow oil.

[0521] 1H-NMR (400MHz, CDCl3) δ7.48-7.46(m,2H),7.41-7.32(m,3H),7.13-7.04(m,2H),6.69(d,J=8Hz,1H),6. 55(s,1H),5.18(s,2H),3.66(s,3H),2.81(t,J=7.2Hz,2H),2.41(t,J=7.2Hz,2H),2.07(p,J=7.2Hz,2H).

[0522] Step 2: Preparation of compound C-10b

[0523] Compound C-10a (894 mg, 2.75 mmol) was dissolved in MC (50 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (748 μL, 8.27 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 8.27 mL, 8.27 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 1.5 hours. After the reaction was completed, distilled water (250 mL) was slowly added dropwise to terminate the reaction, and MC (250 mL) was added to extract the organic layer twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound C-10b (511 mg, 53%) in the form of a yellow oil.

[0524] 1 H-NMR (400MHz, CDCl3) δ10.25(s,1H),7.68(d,J=8.4Hz,1H),7.47-2.27(m,5H),6.81(d,J=8.4Hz,1H),6. 61(s,1H),5.27(s,2H),3.67(s,3H),2.89(t,J=7.2Hz,2H),2.43(t,J=7.2Hz,2H),2.11(p,J=7.2Hz,2H).

[0525] Step 3: Preparation of Core C-10

[0526] Compound C-10b (511 mg, 1.45 mmol) was dissolved in MC (30 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 4.35 mL, 4.35 mmol) was slowly added, and the mixture was stirred for 1.5 hours. After the reaction was completed, distilled water (150 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (150 mL) was added to the mixture, and the organic layer was extracted twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain core C-10 (200.2 mg, 52%) in the form of a white solid.

[0527] 1 H-NMR (400MHz, DMSO-d6) δ10.04(s,1H),7.61(d,J=8.4Hz,1H),6.74(d,J=8.4Hz,1H),6.7 1(s,1H),3.58(s,3H),2.82(t,J=7.2Hz,2H),2.43(t,J=7.2Hz,2H),1.95(p,J=7.2Hz,2H).

[0528] Preparation Example 24: Preparation of Core C-11

[0529]

[0530] Core C-9 (200.2 mg, 0.76 mmol) was dissolved in ethanol (5 mL) and THF (5 mL) at room temperature under a nitrogen atmosphere. Lithium hydroxide monohydrate (64 mg, 1.53 mmol) dissolved in distilled water (2.5 mL) was then slowly added dropwise, and the mixture was stirred at room temperature for 2.5 hours. After the reaction was complete, EA (150 mL) and 2N-hydrochloric acid aqueous solution (150 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain Core C-11 (210 mg, 99%) as a black solid.

[0531] 1 H-NMR (400MHz, DMSO-d6) δ11.08(s,1H),10.05(s,1H),7.61(d,J=8.4Hz,1H),6.74(d,J=8. 4Hz, 1H), 6.71 (s, 1H), 2.82 (t, J = 7.2Hz, 2H), 2.33 (t, J = 7.2Hz, 2H), 1.90 (p, J = 7.2Hz, 2H).

[0532] Preparation Example 25: Preparation of Core C-12

[0533]

[0534] Step 1: Preparation of compound C-12b

[0535] Compound C-12a (10 g, 90.8 mmol) was dissolved in ACN (200 mL) at room temperature under a nitrogen atmosphere, and then benzyl bromide (7.18 mL, 60.5 mmol) was added, and the mixture was cooled to 0 ° C. Potassium carbonate (20.9 g, 151.2 mmol) was added, and the mixture was stirred at room temperature for 20 hours. After the reaction was completed, the organic layer was extracted twice with EA (400 mL) and distilled water (500 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained by column chromatography gave compound C-12b (6.6 g, 54%) as a yellow solid.

[0536] 1 H-NMR (400MHz, CDCl3) δ7.43-7.36(m,4H),7.34-7.32(m,1H),7.13(t,J=8.4Hz,1H),6.56(dd,J =2.4,0.8Hz,1H),6.48(t,J=2.4Hz,1H),6.43(dd,J=2.4,0.8Hz,1H),5.03(s,2H),4.97(s,1H).

[0537] Step 2-1: Preparation of compound C-12c

[0538] Compound C-12b (1.0 g, 4.99 mmol) was dissolved in ACN (25 mL) at room temperature under a nitrogen atmosphere, and then chloromethyl methyl ether (CAS No.: 107-30-2, 7.18 mL, 5.99 mmol) and potassium carbonate (1.37 g, 9.98 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the organic layer was extracted twice with EA (100 mL) and distilled water (100 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to give compound C-12c (908 mg, 74%) as a white solid.

[0539] 1 H-NMR (400MHz, CDCl3) δ7.44-7.42(m,2H),7.39-7.36(m,2H),7.33-7.31(m,1H),7.18(t,J =8.4Hz,1H),6.70-6.69(m,1H),6.66-6.62(m,2H),5.15(s,2H),5.04(s,2H),3.47(s,3H).

[0540] Step 3-1: Preparation of compound C-12d

[0541] Compound C-12c (900 mg, 3.68 mmol) was added to THF (30 mL) at room temperature under a nitrogen atmosphere, and the mixture was cooled to -40 ° C., and then n-butyl lithium solution (2.5 M n-BuLi in hexane, 1.76 mL, 4.42 mmol) was slowly added dropwise. The temperature of the mixture was raised to -20 ° C., and then DMF (0.85 mL) was slowly added dropwise, and the temperature was slowly raised to room temperature, and then the mixture was stirred for 1 hour. After the reaction was completed, distilled water (30 mL) was added dropwise to terminate the reaction. The organic layer was extracted twice from the reaction solution using EA (30 mL), and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained compound C-12d (300 mg, 30%) as a white solid by column chromatography.

[0542] 1 H-NMR (400MHz, CDCl3) δ10.60(s,1H),7.46-7.45(m,2H),7.41-7.34(m,3H),7.32-7.30(m ,1H),6.80(d,J=8.4Hz,1H),6.67(d,J=8.4Hz,1H),5.26(s,2H),5.18(s,2H),3.50(s,3H).

[0543] Step 4-1: Preparation of compound C-12g (Method 1)

[0544] Compound C-12d (300 mg, 1.10 mmol) was dissolved in ethanol (10 mL) at room temperature under a nitrogen atmosphere, and then 2N-hydrochloric acid aqueous solution (1.1 mL) was added, and the mixture was stirred for 16 hours. After the reaction was completed, distilled water (50 mL) was added dropwise to terminate the reaction, and the organic layer was extracted twice with EA (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound C-12g (150 mg, 60%) as a white solid.

[0545] 1 H-NMR (400MHz, CDCl3) δ11.95(s,1H),10.41(s,1H),7.41-7.35(m,6H),6.54(d,J=8.4Hz,1H),6.45(d,J=8.4Hz,1H),5 .14(s,2H),3.88(td,J=10.8,3.2Hz,1H),3.63-3.60(m,1H),2.05-1.98(m,1H),1.97-1.86(m,2H),1.73-1.60(m,3H).

[0546] Step 2-2: Preparation of compound C-12e

[0547] Compound C-12b (22 g, 109.8 mmol) was dissolved in MC (200 mL) under a nitrogen atmosphere at room temperature, and then 3,4-dihydro-2H-pyran (CAS No.: 110-87-2, 12.0 mL, 131.8 mmol) and PPTS (2.76 g, 10.9 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the organic layer was extracted twice with MC (500 mL) and distilled water (500 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound A-24e (23.9 g, 76%) as a white solid.

[0548] 1 H-NMR (400MHz, CDCl3) δ7.44-7.42(m,2H),7.40-7.36(m,2H),7.33-7.29(m,1H ),7.17(t,J=8.4Hz,1H),6.72(t,J=2.4Hz,1H),6.66(dd,J=2.4,0.8Hz,1H),6. 61(dd,J=2.4,0.8Hz,1H),5.39(t,J=3.6Hz,1H),5.04(s,2H),3.94-3.88(m,1H ),3.62-3.57(m,1H),2.04-1.95(m,1H),1.86-1.82(m,2H),1.69-1.53(m,3H).

[0549] Step 3-2: Preparation of compound C-12f

[0550] Compound C-12e (7.7 g, 27.0 mmol) was dissolved in THF (200 mL) at room temperature under a nitrogen atmosphere, and the mixture was cooled to 0 ° C. and then n-butyl lithium solution (2.5 M n-BuLi in hexane, 13.0 mL, 32.4 mmol) was slowly added dropwise. The reaction solution was stirred for 30 minutes, and then DMF (6.3 mL) was slowly added dropwise, slowly warmed to room temperature, and then stirred for another hour. After the reaction was completed, distilled water (500 mL) was added dropwise to terminate the reaction. The organic layer was extracted twice from the reaction solution using EA (500 mL), and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was obtained by column chromatography to give compound C-12f (2.5 g, 27%) as a white solid.

[0551] 1H-NMR (400MHz, CDCl3) δ10.62(s,1H),7.47-7.46(m,2H),7.40-7.36(m,3H),7.35-7.31(m,1H),6.82(d,J=8.4Hz,1H),6.64 (d,J=8.4Hz,1H),3.88(td,J=10.8,3.2Hz,1H),3.63-3.60(m,1H),2.05-1.98(m,1H),1.97-1.86(m,2H),1.73-1.60(m,3H).

[0552] Step 4-2: Preparation of compound C-12g (Method 2)

[0553] Compound C-12f (3.98 g, 12.7 mmol) was dissolved in ethanol (80 mL) at room temperature under a nitrogen atmosphere, and PPTS (2.76 g, 10.9 mmol) was then added. The mixture was stirred at 60 ° C for 2 hours. After the reaction was completed, distilled water (300 mL) was added dropwise to terminate the reaction. EA (300 mL) was added to the reaction solution, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound C-12g (2.6 g, 89%) as a white solid.

[0554] 1 H-NMR (400MHz, CDCl3) δ11.97(s,1H),10.41(s,1H),7.42-7.38(m,6H),6.54(d,J=8.4Hz,1H),6.45(d,J=8.4Hz,1H),5 .14(s,2H),3.88(td,J=10.8,3.2Hz,1H),3.63-3.60(m,1H),2.05-1.98(m,1H),1.97-1.86(m,2H),1.73-1.60(m,3H).

[0555] Step 5: Preparation of compound C-12h

[0556] Compound C-12g (2.5g, 10.9mmol) is dissolved in DMF (50mL) at room temperature under a nitrogen atmosphere, and then ethyl chloroacetate (1.40mL, 13.1mmol) and potassium carbonate (3.02g, 21.9mmol) are added, and the mixture is stirred at 80°C for 2 hours. After the reaction is completed, distilled water (300mL) is added dropwise to terminate the reaction. EA (300mL) is added to the reaction solution to extract the organic layer twice, and sodium chloride aqueous solution is added to the obtained organic layer to extract the organic layer again. The washed organic layer is dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtains compound C-12h (2.7gmg, 77%) as a white solid through column chromatography.

[0557] 1 H-NMR (400MHz, CDCl3) δ10.63(s,1H),7.46-7.45(m,2H),7.40-7.36(m,3H),7.33-7.32(m,1H),6.68(d,J= 8.4Hz,1H),6.45(d,J=8.4Hz,1H),5.18(s,2H),4.72(s,2H),4.26(q,J=7.2Hz,2H),1.28(d,J=7.2Hz,3H).

[0558] Step 6: Preparation of compound C-12i

[0559] Compound C-12h (2.7 g, 8.59 mmol) was dissolved in DMF (50 mL) at room temperature under a nitrogen atmosphere, followed by the addition of potassium carbonate (1.78 g, 12.9 mmol), and the mixture was stirred at 100 ° C for 2 hours. After the reaction was completed, distilled water (200 mL) was added dropwise to terminate the reaction. EA (300 mL) was used to extract the organic layer twice from the reaction solution, and the resulting organic layer was washed with sodium chloride aqueous solution. The washed organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained obtained compound C-12i (2.1 g mg, 82%) as a white solid by column chromatography.

[0560] 1 H-NMR(400MHz, CDCl3)δ7.67(s,1H),7.48-7.46(m,2H),7.43-7.39(m,2H),7.37-7.32(m,2H),7.20 (d,J=8.4Hz,1H),6.75(d,J=8.4Hz,1H),5.21(s,2H),4.42(q,J=7.2Hz,2H),1.42(t,J=7.2Hz,3H).

[0561] Step 7: Preparation of compound C-12j

[0562] Compound C-12i (910 mg, 3.07 mmol) was dissolved in MC (60 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (833 μL, 9.21 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 9.2 mL, 9.21 mmol) were slowly added in sequence, and the mixture was stirred for 2 hours. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound C-12j (270 mg, 26.7%).

[0563] 1 H-NMR (400MHz, CDCl3) δ10.43 (s, 1H), 7.95 (d, J = 8.4Hz, 1H), 7.71 (s, 1H), 7.47-7.38 (m ,5H),6.88(d,J=8.4Hz,1H),5.30(s,2H),4.44(q,J=7.2Hz,2H),1.42(t,J=7.2Hz,3H).

[0564] Step 8: Preparation of Core C-12

[0565] Compound C-12j (250 mg, 0.77 mmol) was dissolved in MC (15 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 4.3 mL, 4.3 mmol) was slowly added, and the mixture was stirred for 1 hour. After the reaction was completed, distilled water (50 mL) was slowly added dropwise at -50 ° C to terminate the reaction. EA (100 mL) and distilled water (100 mL) were added at room temperature, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain core C-12 (150 mg, 83%) as a white solid.

[0566] 1 H-NMR (400MHz, CDCl3) δ10.37(s,1H),7.85(d,J=8.4Hz,1H),7.76(s,1H),6.81(d,J=8.4Hz,1H),4.44(q,J=7.2Hz,2H),1.43(t,J=7.2Hz,3H).

[0567] Preparation Example 26: Preparation of Core C-13

[0568]

[0569] Step 1: Preparation of compound C-13a

[0570] LAH (2.65 g, 69.77 mmol) was added to THF (350 mL) at 0 ° C under a nitrogen atmosphere, and the compound C-12i (8.27 g, 27.91 mmol) prepared in step 6 of Preparation Example 25 was dissolved in THF (50 mL) and slowly added, and the mixture was stirred at 0 ° C for 1 hour. After the reaction was completed, distilled water (300 mL) was added to terminate the reaction, the reaction solution was diluted with EA (300 mL), and filtered with celite. The resulting solution was concentrated under reduced pressure to remove the organic layer, and then EA (500 mL) was added to extract the organic layer twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain C-13a (6.99 g, 98%) in the form of a white solid.

[0571] 1 H-NMR (400MHz, CDCl3) δ7.47-7.45(m,2H),7.41-7.37(m,2H),7.35(m,1H),7.18(t,J=8Hz,1H),7.10( d,J=8Hz,1H),6.80(s,1H),6.71(d,J=8Hz,1H),5.19(s,2H),4.73(d,J=6Hz,2H),1.88(t,J=6Hz,1H).

[0572] Step 2: Preparation of compound C-13b

[0573] Dess-Martin periodinane (CAS No.: 87413-09-0, 7.02 g, 16.56 mmol) was added to MC (25 mL) at 0°C under a nitrogen atmosphere, and then compound C-13 (3.5 g, 13.80 mmol) dissolved in MC (10 mL) was slowly added, and the mixture was stirred at 0°C for 30 minutes. The mixture was stirred for another 3 hours while the temperature was raised to room temperature. After the reaction was completed, the reaction solution was diluted with MC (200 mL) and filtered through diatomaceous earth, followed by addition of distilled water (200 mL) and extraction of the organic layer twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain C-13b (3.31 g, 95%) in the form of a light yellow solid.

[0574] 1 H-NMR (400MHz, CDCl3) δ9.80 (s, 1H), 7.71 (s, 1H), 7.48-7.36 (m, 6H), 7.20 (d, J = 8Hz, 1H), 6.77 (d, J = 8Hz, 1H), 5.22 (s, 2H).

[0575] Step 3: Preparation of compound C-13c

[0576] Compound C-13b (3.31 g, 13.12 mmol) was dissolved in MC (65 mL) at room temperature under a nitrogen atmosphere, followed by the addition of (carboethoxymethylene)triphenylphosphorane (Merck, CAS No. 99-45-2, 9.14 g, 26.24 mmol), and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound C-13c (4.24 g, 99%) as a white solid.

[0577] Cis: 1 H-NMR (400MHz, CDCl3) δ8.03(s,1H),7.48-7.46(m,2H),7.41-7.33(m,3H),7.23(m,1H),7.07(d,J=8Hz,1H),6.85(d,J=1 3.2Hz, 1H), 6.68 (d, J = 8Hz, 1H), 5.92 (d, J = 13.2Hz, 1H), 5.21 (s, 2H), 4.28 (q, J = 7.2Hz, 2H), 1.34 (t, J = 7.2Hz, 3H); EI-MS m / z:323[M+H] + .

[0578] Trans: 1 H-NMR (400MHz, CDCl3) δ7.52(d,J=15.6Hz,1H),7.48-7.45(m,2H),7.42-7.35(m,3H),7.24(m,1H),7.10(d,J=8Hz,1H),7 .07(s,1H),6.71(d,J=8Hz,1H),6.53(d,J=15.6Hz,1H),5.20(s,2H),4.27(q,J=7.2Hz,2H),1.34(t,J=7.2Hz,3H); EI-MS m / z:323[M+H] + .

[0579] Step 4: Preparation of compound C-13d

[0580] Compound C-13c (4.24 g, 13.15 mmol) was dissolved in THF (160 mL) and methanol (480 mL) at room temperature under a nitrogen atmosphere, and 5% palladium on carbon (5% Pd / C, 2.1 g) was added, and the mixture was stirred for 3 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was diluted with EA (350 mL) and filtered through diatomaceous earth. The filtered solution was concentrated under reduced pressure. The resulting residue obtained was obtained by column chromatography as a white solid compound C-13d (2.93 g, 95%).

[0581] 1 H-NMR (400MHz, CDCl3) δ7.08-7.00(m,2H),6.59(dd,J=7.6,0.8Hz,1H),6.49(d,J=0.8Hz,1H),5.41(s ,1H),4.16(q,J=7.2Hz,2H),3.10(t,J=7.2Hz,2H),2.75(t,J=7.2Hz,2H),1.26(t,J=7.2Hz,3H); EI-MS m / z:235[M+H] + .

[0582] Step 5: Preparation of compound C-13e

[0583] Compound C-13d (2.93 g, 12.51 mmol) was dissolved in ACN (100 mL) at room temperature under a nitrogen atmosphere, followed by the addition of potassium carbonate (4.3 g, 31.27 mmol) and benzyl bromide (1.9 mL, 16.26 mmol), and the mixture was stirred at 80° C. for 16 hours. After the reaction was complete, EA (500 mL) and distilled water (450 mL) and 2N-hydrochloric acid aqueous solution (50 mL) were added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained was subjected to column chromatography to obtain compound C-13e (3.99 g, 98%) as a colorless oil.

[0584] 1 H-NMR (400MHz, CDCl3) δ7.47-7.45(m,2H),7.41-7.37(m,2H),7.33(m,1H),7.12(t,J=8Hz,1H),7.05(d,J=8Hz,1H),7.68(d,J=8H EI-MS m / z:325[M+H] + .

[0585] Step 6: Preparation of compound C-13f

[0586] Compound C-13e (3.99 g, 12.3 mmol) was dissolved in MC (240 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (3.3 mL, 36.9 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 36.9 mL, 36.9 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 1.5 hours. After the reaction was completed, distilled water (500 mL) was slowly added dropwise to terminate the reaction, and MC (350 mL) was added to extract the organic layer twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound C-13f (2.3 g, 53%) as a yellow solid.

[0587] 1 H-NMR (400MHz, CDCl3) δ10.24(s,1H),7.68(d,J=8.4Hz,1H),7.46-7.34(m,5H),6.80(d,J=8.4Hz,1H),6.62(s ,1H),5.26(s,2H),4.17(q,J=7.2Hz,2H),3.18(t,J=7.2Hz,2H),2.78(t,J=7.2Hz,2H),1.25(t,J=7.2Hz,3H).

[0588] Step 7: Preparation of Core C-13

[0589] Compound C-13f (2.3 g, 6.53 mmol) was dissolved in ethanol (40 mL) and THF (40 mL) at 0°C under a nitrogen atmosphere, and then lithium hydroxide monohydrate (821 mg, 19.58 mmol) dissolved in distilled water (20 mL) was slowly added dropwise, and the mixture was stirred at room temperature for 1.5 hours. After the reaction was completed, the organic layer was extracted with EA (300 mL) and 2N-hydrochloric acid aqueous solution (200 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain core C-13 (2.4 g, 99%) in the form of a green solid.

[0590] 1 H-NMR(400MHz,DMSO-d6)δ10.11(s,1H),7.76(d,J=8.4Hz,1H),7.53-7.51(m,2H),7.44-7.36(m,3 H),7.08(d,J=8.4Hz,1H),6.74(s,1H),5.37(s,2H),3.04(t,J=7.2Hz,2H),2.71(t,J=7.2Hz,2H).

[0591] Preparation Example 27: Preparation of Core C-14

[0592]

[0593] The compound C-13b (1 g, 3.96 mmol) prepared in step 2 of Preparation Example 26 was dissolved in MC (80 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 11.89 mL, 11.89 mmol) was slowly added, and the mixture was stirred at -78 ° C for 2 hours. After the reaction was completed, distilled water (250 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (250 mL) was added to the mixture and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue obtained by column chromatography obtained core C-14 (580 mg, 90%) in the form of a yellow solid.

[0594] 1 H-NMR (400MHz, DMSO-d6) δ10.61(brs,1H),9.78(s,1H),7.97(s,1H),7.38(t,J=8.4Hz,1H),7.12(d,J=8.4Hz,1H),6.71(d,J=8.4Hz,1H).

[0595] Preparation Example 28: Preparation of Core C-15

[0596]

[0597] Step 1: Preparation of compound C-15b

[0598] Compound C-15a (2-chloro-6-hydroxybenzaldehyde, Merck, CAS No.: 18362-30-6, 500 mg, 3.19 mmol) was dissolved in ACN (10 mL) at 0°C under a nitrogen atmosphere, followed by the addition of benzyl bromide (400 μL, 3.35 mmol) and potassium carbonate (1.1 g, 7.98 mmol), and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, EA (100 mL) and distilled water (100 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to give compound C-15b (750 mg, 95%) as a white solid.

[0599] 1H-NMR (400MHz, CDCl3) δ10.57 (s, 1H), 7.45-7.34 (m, 6H), 7.04 (d, J = 8Hz, 1H), 6.96 (d, J = 8.8Hz, 1H), 5.19 (s, 2H).

[0600] Step 2: Preparation of compound C-15c

[0601] Selenium powder (256 mg, 3.24 mmol) was added to THF (10 mL) at room temperature under a nitrogen atmosphere, the mixture was cooled to 0 ° C, and then n-butyl lithium solution (2.5 M n-BuLi in hexane, 1.42 mL, 3.56 mmol) was slowly added dropwise. The mixture was stirred at 0 ° C for 40 minutes, and then compound C-15b (800 mg, 3.24 mmol) dissolved in DMF (2 mL) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled to 0 ° C and distilled water (100 mL) was slowly added dropwise to terminate the reaction. EA (100 mL) was added to the reaction solution, and the reaction solution was then extracted, and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained by column chromatography gave compound C-15c (900 mg, 80%) as a light yellow solid.

[0602] 1 H-NMR (400MHz, CDCl3) δ10.70(s,1H),7.42-7.33(m,6H),7.04(d,J=8Hz,1H),6.81(d,J=8.8Hz,1H ),5.18(s,2H),2.85(t,J=7.6Hz,2H),1.75(qui,J=7.6Hz,2H),1.51(m,2H),0.95(t,J=7.6Hz,3H).

[0603] Step 3: Preparation of compound C-15d

[0604] Compound C-15c (900 mg, 2.59 mmol) was dissolved in DMF (10 mL) at room temperature under a nitrogen atmosphere, and then ethyl bromoacetate (574 μL, 6.48 mmol) was added, and the mixture was stirred at 120° C. for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature and used in the next reaction without additional purification.

[0605] Step 4: Preparation of compound C-15e

[0606] Potassium carbonate (716 mg, 6.48 mmol) was added to the reaction solution under a nitrogen atmosphere, and the mixture was stirred at 120 ° C for 2.5 hours. After the reaction was completed, EA (100 mL) and distilled water (200 mL) were added to extract the organic layer. The organic layer was further washed by adding distilled water (200 mL) to the obtained organic layer again, and the organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained by column chromatography gave compound C-15e (840 mg, 90%) as a white solid.

[0607] 1 H-NMR(400MHz, CDCl3)δ8.55(s,1H),7.49-7.47(m,3H),7.43-7.29(m,4H),6.84 (d, J=8Hz, 1H), 5.21 (s, 2H), 4.37 (q, J=7.2Hz, 2H), 1.39 (t, J=7.2Hz, 3H); EI-MS m / z:361[M+H] + .

[0608] Step 5: Preparation of compound C-15f

[0609] Compound C-15e (400 mg, 1.11 mmol) was dissolved in MC (30 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (300 μL, 3.33 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 3.33 mL, 3.33 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 2 hours. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound C-15f (310 mg, 72%).

[0610] 1 H-NMR (400MHz, CDCl3) δ10.09 (s, 1H), 8.62 (s, 1H), 7.89 (d, J = 8Hz, 1H), 7.49-7.38 (m, 5 H),7.03(d,J=8Hz,1H),5.34(s,2H),4.39(q,J=7.2Hz,2H),1.41(t,J=7.2Hz,3H); EI-MS m / z:388[M+H] + .

[0611] Step 6: Preparation of Core C-15

[0612] Compound C-15f (300 mg, 0.77 mmol) was dissolved in MC (15 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 4.62 mL, 4.62 mmol) was slowly added, and the mixture was stirred for 1.5 hours. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain core C-15 (185 mg, 80%) as a white solid.

[0613] 1 H-NMR (400MHz, DMSO-d6) δ10.05(s,1H),8.47(s,1H),8.09(d,J=8Hz,1H),7.05(d,J=8Hz,1H),4.33(q,J=7.2Hz,2H),1.34(t,J=7.2Hz,3H).

[0614] Preparation Example 29: Preparation of Core C-16

[0615]

[0616] Compound C-2d (3.0 g, 10.15 mmol) was dissolved in MC (180 mL) at -30°C under a nitrogen atmosphere, followed by the slow addition of dichloromethyl methyl ether (Merck, CAS No. 4885-02-3, 2.76 mL, 30.45 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 30.45 mL, 30.45 mmol), and the mixture was stirred for 2 hours. After the reaction was complete, cold distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (300 mL) was added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain core C-16 (2.76 g, 84%).

[0617] 1 H-NMR (400MHz, CDCl3) δ10.07(S,1H),9.28(brs,1H),7.94(d,J=2.0Hz,1H),7.61(d,J=8Hz,1H), 7.49-7.40(m,5H),6.90(d,J=8Hz,1H),5.31(s,2H),4.41(q,J=7.2Hz,2H),1.41(t,J=7.2Hz,3H).

[0618] Preparation Example 30: Preparation of Linker Q-1

[0619]

[0620] Linker P-2 (172 mg, 0.64 mmol) was dissolved in a saturated aqueous sodium bicarbonate solution (3.5 mL) at 0°C under a nitrogen atmosphere, and the mixture was stirred at 0°C for 15 minutes. Q-1a (N-methoxycarbonylmaleimide, TCI, CAS No.: 55750-48-6, 100 mg, 0.64 mmol) was slowly added to the mixture, and the mixture was stirred at the same temperature for 1.5 hours. After the reaction was completed, EA (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure at 20°C or lower. The resulting residue was subjected to column chromatography to obtain linker Q-1 (137.3 mg, 68%) as a colorless oil.

[0621] 1 H-NMR (400MHz, CDCl3) δ6.70 (s, 2H), 4.20 (d, J = 2.4Hz, 2H), 3.74-3.59 (m, 16H), 2.42 (t, J = 2.4Hz, 1H).

[0622] Preparation Example 31: Preparation of Linker Q-2

[0623]

[0624] Compound Q-2a (Mal-PEG2-acid, CAS No.: 1374666-32-6, TCI, 100 mg, 0.388 mmol) was dissolved in MC (5 mL) at 0 ° C under a nitrogen atmosphere, followed by the addition of NHS (49.2 mg, 0.427 mmol) and DCC (88.2 mg, 0.427 mmol), and the mixture was stirred for 15 hours. After the reaction was complete, EA / n-hexane (1: 1 volume ratio, 20 mL) was added and the resulting precipitate was removed by filtration. The filtrate was concentrated, and then EA / n-hexane (1: 1 volume ratio, 20 mL) was added again to remove the resulting precipitate. The filtrate was concentrated under reduced pressure to give connector Q-2 (139 mg, 99%).

[0625] 1 H-NMR (400MHz, CDCl3) δ6.70 (s, 2H), 3.81 (t, J=6.4Hz, 2H), 3.72 (m, 2H), 3.65-3.58 (m, 6H), 2.87 (t, J=6.4Hz, 2H), 2.84 (s 4H); EI-MS m / z: 355[M+H] + .

[0626] Preparation Example 32: Preparation of Linker Q-3

[0627]

[0628] Step 1: Preparation of compound Q-3b

[0629] Compound Q-3a (diethanolamine, Daejung Chemicals & Metals, CAS No.: 111-42-2, 10 g, 95.12 mmol) was dissolved in 1,4-dioxane (320 mL) at 0°C under a nitrogen atmosphere, and then sodium bicarbonate (16 g, 190.24 mmol) and di-tert-butyl dicarbonate (Boc anhydride, 25 g, 114.14 mmol) dissolved in distilled water (160 mL) were added sequentially, and the mixture was stirred for 48 hours while the temperature was slowly raised from 0°C to room temperature. After the reaction was completed, EA (500 mL), distilled water (400 mL), and 2N-hydrochloric acid aqueous solution (100 mL) were added, and the organic layer was extracted five times. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain compound Q-3b (17.89 g, 91%) as a colorless liquid.

[0630] 1 H-NMR (400MHz, DMSO-d6) δ4.67(brs,2H),3.49-3.42(m,4H),3.25-3.19(m,4H),1.38(s,9H).

[0631] Step 2: Preparation of compound Q-3c

[0632] Compound Q-3b (1.02 g, 4.97 mmol) was dissolved in DMF (16 mL) at 0°C under a nitrogen atmosphere, and then propargyl bromide (2.2 mL, 19.88 mmol) and potassium hydroxide (1.11 g, 19.88 mmol) were added, and the mixture was stirred for 16 hours while slowly raising the temperature from 0°C to room temperature. After the reaction was completed, EA (250 mL), distilled water (150 mL) and 2N-hydrochloric acid aqueous solution (100 mL) were added, and the organic layer was extracted twice. The obtained organic layer was washed by adding saturated sodium chloride aqueous solution (250 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound Q-3c (806 mg, 57%) as a light yellow liquid.

[0633] 1 H-NMR (400MHz, CDCl3) δ4.14 (d, J = 2Hz, 4H), 3.67-3.61 (m, 4H), 3.51-3.43 (m, 4H), 2.43-2.40 (m, 2H), 1.46 (s, 9H).

[0634] Step 3: Preparation of compound Q-3d

[0635] Compound Q-3c (806 mg, 2.86 mmol) was dissolved in dichloromethane (5 mL) at 0°C under a nitrogen atmosphere, and then a 4N-hydrochloric acid solution (4M-HCl in dioxane, 10 mL) was added, and the mixture was stirred at 0°C for 1 hour, and then the mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain compound Q-3d (618 mg, 99%) as an ivory solid.

[0636] 1 H-NMR (400MHz, CDCl3) δ9.46 (brs, 1H), 4.26 (d, J = 2.4Hz, 4H), 3.98 (t, J = 5.2Hz, 4H), 3.38-3.30 (m, 4H), 2.50 (t, J = 2.4Hz, 2H).

[0637] Step 4: Preparation of compound Q-3e

[0638] Compound Q-3d (618 mg, 2.84 mmol) and linker P-8 (747 mg, 2.84 mmol) were dissolved in DMF (15 mL) at 0°C under a nitrogen atmosphere, and then EDCI hydrochloride (816 mg, 4.26 mmol), HOBt (384 mg, 2.84 mmol), DMAP (35 mg, 0.28 mmol) and DIPEA (1.48 mL, 8.51 mmol) were added in sequence, and the mixture was stirred for 16 hours while the temperature was slowly raised from 0°C to room temperature. After the reaction was completed, the organic layer was extracted twice with EA (150 mL), distilled water (100 mL) and 2N-hydrochloric acid aqueous solution (50 mL). Saturated sodium chloride aqueous solution (200 mL) was added to the obtained organic layer and the organic layer was washed. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound Q-3e (887 mg, 73%) as a yellow oil.

[0639] 1 H-NMR(400MHz, CDCl3)δ5.08(brs,1H),4.30(s,2H),4.16-4.11(m,4H),3.73-3.68(m,4H),3.68-3.63( EI-MS m / z:427(M + ).

[0640] Step 5: Preparation of compound Q-3f

[0641] Compound Q-3e (887 mg, 2.08 mmol) was dissolved in dichloromethane (4 mL) at 0°C under a nitrogen atmosphere, and then a 4N-hydrochloric acid solution (4M-HCl in dioxane, 8 mL) was added, and the mixture was stirred at 0°C for 1 hour, and then the mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain compound Q-3f (824 mg, quantitatively obtained) as an ivory solid.

[0642] 1 H-NMR (400MHz, CDCl3) δ8.42(brs,2H),4.35(s,2H),4.18-4.14(m,4H),3.92(t,J=4.8Hz,2H),3.76-3.73(m,2H EI-MS m / z:327[M+H] + .

[0643] Step 6: Preparation of compound Q-3

[0644] Compound Q-3f (147 mg, 0.40 mmol) was dissolved in a saturated aqueous sodium bicarbonate solution (sodium bicarbonate, 2 mL) at 0°C under a nitrogen atmosphere, and the mixture was stirred at 0°C for 15 minutes. Q-1a (N-methoxycarbonylmaleimide, TCI, CAS No.: 55750-48-6, 63 mg, 0.40 mmol) was slowly added to the mixture, and the mixture was stirred at the same temperature for 1.5 hours. After the reaction was completed, EA (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted three times. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure at 15°C or lower. The resulting residue was subjected to column chromatography to give compound Q-3 (55.1 mg, 33%) as a colorless oil.

[0645] 1 H-NMR(400MHz, CDCl3)δ6.69(s,2H),4.27(s,2H),4.16-4.11(m,4H),3.75- 3.68(m,4H),3.67-3.62(m,8H),3.61-3.57(m,4H),2.45-2.41(m,2H); EI-MS m / z:407[M+H] + .

[0646] Preparation Example 33: Preparation of Linker Q-4

[0647]

[0648] Step 1: Preparation of compound Q-4a

[0649] Compound P-1a (tetraethylene glycol, Daejung Chemicals & Metals, CAS No. 112-60-7, 5 g, 25.74 mmol) was dissolved in THF (60 mL) at room temperature under a nitrogen atmosphere. Sodium hydride (NaH 60% dispersion in mineral oil, 16.5 mg, 0.41 mmol) was then added, and the mixture was stirred at room temperature for 30 minutes. Tert-butyl acrylate (Merck, CAS No. 1663-39-4, 1.5 mL, 10.30 mmol) was slowly added to the reaction solution over 2 hours, and the mixture was stirred at room temperature for 3 hours. After completion, saturated aqueous sodium chloride solution (20 mL) was added to terminate the reaction, and the reaction solvent was removed by concentration under reduced pressure. MC (60 mL) was added to the remaining aqueous sodium chloride solution to extract the organic layer, which was then dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound Q-4a (1.39 g, 42%) as a pink oil.

[0650] 1 H-NMR (400MHz, CDCl3) δ3.75-3.60(m,18H),2.69(t,J=6.4Hz,1H),2.50(t,J=6.4Hz,2H),1.45(s,9H); EI-MS m / z:345[M+Na] + .

[0651] Step 2: Preparation of compound Q-4b

[0652] Compound Q-4a (1.39 g, 4.31 mmol) was dissolved in ACN (7.5 mL) at 0°C under a nitrogen atmosphere, and pyridine (4 mL) was slowly added. A solution of 4-methylbenzenesulfonyl chloride (p-toluenesulfonyl chloride, 1.1 g, 5.77 mmol) dissolved in ACN (10.5 mL) was then slowly added over 30 minutes, and the mixture was stirred at room temperature for 6.5 hours. After the reaction was complete, EA (200 mL) and distilled water (150 mL) and 2N-hydrochloric acid aqueous solution (50 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained was obtained by column chromatography as a colorless oil compound Q-4b (1.78 g, 86%).

[0653] 1H-NMR (400MHz, CDCl3) δ7.80 (d, J = 8Hz, 2H), 7.34 (d, J = 8Hz, 2H), 4.16 (t, J = 4.8Hz, 2H), 3.7 3-3.65(m,4H),3.65-3.57(m,12H),2.50(t,J=6.4Hz,2H),2.45(s,3H),1.44(s,9H); EI-MS m / z:499[M+Na] + .

[0654] Step 3: Preparation of compound Q-4c

[0655] Compound Q-4b (1.78 g, 3.73 mmol) was dissolved in DMF (20 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (NaN3, 364 mg, 5.60 mmol) was added, and the mixture was stirred at 60 ° C for 16 hours. After the reaction was completed, the mixture was cooled to room temperature, EA (200 mL) and distilled water (200 mL) were added, and the organic layer was extracted twice. The resulting organic layer was washed three times by adding saturated sodium chloride aqueous solution (200 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to give compound Q-4c (1.25 g, 96%) as a colorless oil.

[0656] 1 H-NMR (400MHz, CDCl3) δ3.73-3.60(m,16H),3.39(t,J=4.8Hz,2H),2.50(t,J=6.4Hz,2H),1.44(s,9H); EI-MS m / z:348[M+H] + .

[0657] Step 4: Preparation of compound Q-4d

[0658] Compound Q-4c (500 mg, 1.44 mmol) was dissolved in 1,4-dioxane (8 mL) at room temperature under a nitrogen atmosphere, and then 5% palladium on carbon (5% Pd / C, 153 mg, 0.07 mmol) was added, and the mixture was stirred at 60 ° C for 4 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was diluted with EA (50 mL) and filtered using a diatomaceous earth filter, and the solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound Q-4d (92.5 mg, 20%) as a colorless oil.

[0659] 1H-NMR (400MHz, CDCl3) δ3.71(t,J=6.4Hz,4H),3.65-3.56(m,28H),2.81(t,J=5.6Hz,4H),2.50(t,J=6.4Hz,4H),1.44(s,18H); EI-MS m / z:626[M+H] + .

[0660] Step 5: Preparation of compound Q-4e

[0661] Compound Q-4d (92.5 mg, 0.15 mmol) was dissolved in MC (2 mL) at 0°C under a nitrogen atmosphere, and then linker Q-2 (68.1 mg, 0.19 mmol), pyridine (11.9 μL), and DIPEA (5.1 μL) were added dropwise in that order, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, compound Q-4e (83.6 mg, 65%) was obtained as an orange oil by column chromatography without extraction.

[0662] 1 H-NMR (400MHz, CDCl3) δ6.70 (s, 2H), 3.76-3.69 (m, 8H), 3.65-3.54 (m, 38H), 2.66 (t, J = 6.8Hz, 2H), 2.50 (t, J = 6.4Hz, 4H), 1.44 (s, 18H); EI-MS m / z:866[M+H] + .

[0663] Step 6: Preparation of compound Q-4f

[0664] Compound Q-4e (92.5 mg, 0.15 mmol) was dissolved in MC (3 mL) at 0° C. under a nitrogen atmosphere, and then TFA (0.3 mL) was slowly added, and the mixture was stirred for 6 hours while slowly raising the temperature from 0° C. to 15° C. After the reaction was completed, MC (20 mL) was added to dilute the mixture, and then concentrated under reduced pressure at low temperature to give compound Q-4f (41.2 mg, 99%) as a colorless oil.

[0665] 1 H-NMR (400MHz, CDCl3) δ6.71 (s, 2H), 3.80-3.70 (m, 8H), 3.67-3.54 (m, 38H), 2.71 (t, J = 6.8Hz, 2H), 2.63-2.58 (m, 4H); EI-MS m / z: 753[M+H] + .

[0666] Step 7: Preparation of compound Q-4

[0667] Compound Q-4f (40.5 mg, 0.054 mmol) was dissolved in MC (3 mL) at 0 ° C under a nitrogen atmosphere, and then NHS (13 mg, 0.113 mmol) and DCC (24.4 mg, 0.118 mmol) were added, and the mixture was stirred for 16 hours while slowly raising the temperature from 0 ° C to room temperature. After the reaction was completed, EA / normal hexane (1: 1 volume ratio, 20 mL) was added and the precipitate was removed by filtration. The filtrate was concentrated, and then EA / normal hexane (1: 1 volume ratio, 20 mL) was added again to remove the precipitate. The filtrate was concentrated under reduced pressure to give joint Q-4 (17.4 mg, 43%).

[0668] 1 H-NMR (400MHz, CDCl3) δ6.70 (s, 2H), 3.85 (t, J = 6.4Hz, 4H), 3.76-3.69 (m, 4H), 3.66- 3.52(m,38H),2.90(t,J=6.4Hz,4H),2.85-2.80(m,8H),2.66(t,J=6.8Hz,2H); EI-MS m / z:947[M+H] + .

[0669] Preparation Example 34: Preparation of Linker Q-5

[0670]

[0671] Linker P-1 (164 mg, 0.64 mmol) was dissolved in a saturated aqueous sodium bicarbonate solution (3.5 mL) at 0°C under a nitrogen atmosphere, and the mixture was stirred at 0°C for 20 minutes. Q-1a (N-methoxycarbonylmaleimide, TCI, CAS No. 55750-48-6, 100 mg, 0.64 mmol) was slowly added to the mixture, and the mixture was stirred at the same temperature for 1.5 hours. EA (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted three times. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure at 10°C or lower. The resulting residue was subjected to column chromatography to obtain linker Q-5 (77 mg, 40%) as a colorless oil.

[0672] 1 H-NMR (400MHz, CDCl3) δ6.70 (s, 2H), 3.76-3.71 (m, 2H), 3.69-3.60 (m, 12H), 3.39 (t, J = 5.2Hz, 2H).

[0673] Preparation Example 35: Preparation of Linker Q-6

[0674]

[0675] Step 1: Preparation of compound Q-6a

[0676] Linker Q-2 (70 mg, 0.198 mmol) was dissolved in THF (3 mL) at 0°C under a nitrogen atmosphere, and then 3,3'-iminodipropionic acid (TCI, CAS No.: 505-47-5, 28.6 mg, 0.177 mmol), distilled water (300 μL), and DIPEA (41.3 μL) were added dropwise in sequence, and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the reaction solution was diluted with ACN (1 mL) containing 0.1% formic acid and distilled water (1 mL), and then separated and purified by preparative HPLC, and then lyophilized to obtain compound Q-6a (3 mg, 4%); MS m / z: 401 [M+H] + .

[0677] Step 2: Preparation of compound Q-6

[0678] Compound Q-6a (3 mg, 0.0074 mmol) was dissolved in MC (1 mL) at 0 ° C under a nitrogen atmosphere, and then NHS (1.9 mg, 0.0148 mmol) and DCC (3.24 mg, 0.0155 mmol) were added, and the mixture was stirred for 2 hours. After the reaction was completed, EA / n-hexane (1: 1 volume ratio, 20 mL) was added and the resulting precipitate was removed by filtration. The filtrate was concentrated, and then EA / n-hexane (1: 1 volume ratio, 20 mL) was added again to remove the resulting precipitate. The filtrate was concentrated under reduced pressure to give connector Q-6 (4.5 mg, 99%); EI-MS m / z: 595 [M+H] + .

[0679] Preparation Example 36: Preparation of Linker Q-7

[0680]

[0681] Step 1: Preparation of compound Q-7a

[0682] Compound P-1a (20 g, 102.97 mmol) was dissolved in ethanol THF (80 mL) at 0°C under a nitrogen atmosphere, and sodium hydride (NaH, 60% dispersion in mineral oil, 823 mg, 34.3 mmol) was added, and the mixture was stirred at 0°C for 20 minutes. Propargyl bromide (1.96 mL, 20.6 mmol) was added, and the mixture was stirred at 0°C for 1 hour, and then the mixture was stirred at room temperature for 16 hours. After the reaction was completed, it was extracted with EA (100 mL) and distilled water (100 mL) and saturated sodium chloride aqueous solution (50 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue gave compound Q-7a (3.5 g, 74%) in the form of a yellow oil by column chromatography.

[0683] 1 H-NMR (400MHz, CDCl3) δ4.20 (d, J = 2.0Hz, 2H), 3.72-3.67 (m, 15H), 3.62-3.60 (m, 2H), 2.51 (s, 1H), 2.42 (t, J = 2.0Hz, 1H).

[0684] Step 2: Preparation of compound Q-7b

[0685] Compound Q-7a (1.0 g, 4.30 mmol) was dissolved in MC (50 mL) at room temperature under a nitrogen atmosphere, and then DMP (Dess-Martin periodinane, 2.73 g, 6.45 mmol) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, it was extracted with MC (100 mL), sodium thiosulfate aqueous solution (50 mL) and sodium bicarbonate aqueous solution (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound Q-7b (570 mg, 57%) as a colorless oil.

[0686] 1 H-NMR (400MHz, CDCl3) δ9.73 (s, 1H), 4.20 (d, J = 2.0Hz, 2H), 4.15 (s, 2H), 3.74-3.66 (m, 12H), 2.42 (t, J = 2.0Hz, 1H).

[0687] Step 3: Preparation of compound Q-7c

[0688] Compound Q-7b (1.23 g, 5.34 mmol) was dissolved in methanol (100 mL) at room temperature under a nitrogen atmosphere, followed by the addition of compound P-2 (1.23 g, 5.34 mmol) and acetic acid (0.3 mL, 5.34 mmol), and the mixture was stirred for 10 minutes. Sodium cyanoborohydride (671 mg, 10.68 mmol) was added, and the mixture was stirred for 3 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. The residue was subjected to column chromatography to give compound Q-7c (90 mg, 3.8%) as a colorless oil.

[0689] 1 H-NMR (400MHz, CDCl3) δ4.20 (d, J = 2.0Hz, 4H), 3.78-3.66 (m, 29H), 2.87 (t, J = 5.2Hz, 4H), 2.43 (t, J = 2.0Hz, 2H); EI-MS m / z: 446 [M+H] + .

[0690] Step 4: Preparation of compound Q-7

[0691] Compound Q-7c (40 mg, 0.089 mmol) was dissolved in MC (5 mL) at room temperature under a nitrogen atmosphere, and then compound Q-2 (47.7 mg, 0.134 mmol), DIPEA (7.7 μL, 0.044 mmol) and pyridine (7.3 μL, 0.089 mmol) were added in sequence, and the mixture was stirred at room temperature for 4 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound Q-7 (10.9 mg, 17%) as a colorless oil.

[0692] Example I-1: Preparation of Compounds A-1 and A-2

[0693]

[0694] Step 1: Preparation of compound A-1a

[0695] Core C-16 (1.76 g, 5.46 mmol) was dissolved in DMF (27 mL) at room temperature under a nitrogen atmosphere, followed by the addition of linker P-3 (1.8 g, 6.38 mmol) and potassium carbonate (2.26 g, 16.38 mmol), and the mixture was stirred at 80°C for 16 hours. After the reaction was complete, EA (250 mL) and 2N aqueous hydrochloric acid solution (250 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound A-1a (2.86 g, 99%) as a yellow liquid.

[0696] 1 H-NMR (400MHz, CDCl3) δ10.05(s,1H),8.04(s,1H),7.57(d,J=8Hz,1H),7.50-7.36(m,5H),6.90(d,J=8Hz,1H),5.17-5.13(m,2H) ,4.36(q,J=7.2Hz,2H),3.71-3.67(m,2H),3.59(t,J=4.8Hz,2H),3.53-3.38(m,8H),3.33(t,J=5.2Hz,2H),1.41(t,J=7.2Hz,3H).

[0697] Step 2: Preparation of compound A-1b

[0698] Compound A-1a (2.83 g, 5.39 mmol) was dissolved in MC (95 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 43.15 mL, 43.15 mmol) was slowly added, and the mixture was stirred for 8 hours while slowly raising the temperature to -50 ° C. After the reaction was completed, distilled water (300 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (300 mL) was added to the mixture and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue obtained by column chromatography obtained compound A-1b (1.68 mg, 71%) in the form of a brown liquid.

[0699] 1H-NMR (400MHz, CDCl3) δ10.03(s,1H),9.24(brs,1H),8.12(s,1H),7.56(d,J=8Hz,1H),6.94(d,J=7.6Hz,1H),4.95-4.92(m,2H),4.37(q ,J=7.2Hz,2H),4.15-4.09(m,2H),3.73-3.71(m,2H),3.61-3.58(m,4H),3.57-3.51(m,4H),3.33(t,J=5.2Hz,2H),1.42(t,J=7.2Hz,3H).

[0700] Step 3: Preparation of compound A-1c

[0701] Compound A-1b (1.68 g, 3.87 mmol) was dissolved in MC (130 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (183 mg, 0.44 mmol) and benzyltributylammonium chloride (1.2 g, 3.87 mmol) were added. 5N-sodium hydroxide aqueous solution (2.32 mL, 11.60 mmol) was slowly added to the reaction solution, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, MC (150 mL) and distilled water (150 mL) were added, and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-1c (1.9 g, 64%) in the form of a brown viscous gel.

[0702] 1 H-NMR (400MHz, CDCl3) δ10.10(s,1H),8.02(s,1H),7.57(d,J=8.4Hz,1H),6.97(d,J=8.4Hz,1H),5. 59(dd,J=10.4,8Hz,1H),5.50(d,J=3.2Hz,1H),5.43(d,J=8Hz,1H),5.21-5.17(m,2H),5.01(m,1H) ,4.37(q,J=7.2Hz,2H),4.23-4.11(m,3H),3.69-3.67(m,2H),3.58(t,J=5.2Hz,2H),3.50-3.47(m, 2H),3.43-3.32(m,8H),2.21(s,3H),2.19(s,3H),2.14(s,3H),2.04(s,3H),1.42(t,J=7.2Hz,3H).

[0703] Step 4: Preparation of compound A-1d

[0704] Compound A-1c (1.9 g, 2.49 mmol) was dissolved in THF (40 mL) at 0°C under a nitrogen atmosphere, followed by the addition of sodium borohydride (188.6 mg), and the mixture was stirred for 2.5 hours. After completion of the reaction, distilled water (200 mL) was added to terminate the reaction, and the organic layer was extracted twice with EA (200 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound A-1d (1.55 g, 81%) as a yellow solid.

[0705] 1 H-NMR (400MHz, CDCl3) δ7.38 (s, 1H), 7.01 (d, J = 7.6Hz, 1H), 6.86 (d, J = 8Hz, 1H), 5.55 (dd, J = 10.4, 8Hz, 1H),5.48(d,J=3.2Hz,1H),5.31(d,J=8Hz,1H),5.21-5.14(m,2H),5.01-4.95(m,1H),4.90(d,J=5.2Hz, 2H),4.35(q,J=7.2Hz,2H),4.23-4.09(m,3H),3.71-3.66(m,2H),3.58(t,J=5.2Hz,2H),3.51-3.34(m, 10H),2.21(s,3H),2.07(s,3H),2.05(s,3H),2.02(s,3H),1.76(t,J=5.6Hz,1H),1.41(t,J=7.2Hz,3H).

[0706] Step 5: Preparation of compound A-1e

[0707] Compound A-1d (160 mg, 0.2 mmol) was dissolved in DMF (4 mL) at 0 ° C. under a nitrogen atmosphere, and then bis(4-nitrophenyl) carbonate (127 mg, 0.4 mmol) and DIPEA (54.5 μL, 0.3 mmol) were added in sequence, and the mixture was stirred for 1 hour. The reaction mixture was stirred at room temperature for another 5 hours, and then extracted with EA (50 mL) and distilled water (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-1e (162 mg, 83%) as a light yellow oil.

[0708] 1H-NMR (400MHz, CDCl3) δ8.27(d,J=9.2Hz,2H),7.37(d,J=9.2Hz,2H),7.35(s,1H),7.13(d,J=8Hz,1H),6.87(d ,J=8Hz,1H),5.56(dd,J=10.4,8Hz,1H),5.52(s,2H),5.49(d,J=3.2Hz,1H),5.34(d,J=8Hz,1H),5.21-5.15(m, 2H),5.03-4.98(m,1H),4.38(q,J=7.2Hz,2H),4.24-4.10(m,3H),3.71-3.68(m,2H),3.60(t,J=5.2Hz,2H),3. 54-3.42(m,8H),3.35(m,2H),2.21(s,3H),2.07(s,3H),2.05(s,3H),2.03(s,3H),1.42(t,J=7.2Hz,3H); EI-MS m / z:795[M+H] + .

[0709] Step 6: Preparation of Compound A-1f

[0710] Compound A-1e (30 mg, 0.032 mmol) was dissolved in DMF (0.2 mL) at room temperature under a nitrogen atmosphere, and then MMAF-OMe (24 mg, 0.032 mmol), HOBt (6.5 mg, 0.048 mmol) and DIPEA (14 μL, 0.08 mmol) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC. The product was lyophilized to obtain compound A-1f (12.1 mg, 24%) as a white solid; EI-MS m / z: 1539 [M+H] + .

[0711] Step 7: Preparation of Compounds A-1 and A-2

[0712] Compound A-1f (12.1 mg, 0.008 mmol) was dissolved in methanol (1 mL) at 0°C, and then lithium hydroxide monohydrate (4.3 mg, 0.1 mmol) dissolved in distilled water (250 μL) was slowly added dropwise, and the mixture was stirred at room temperature for 1.5 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC. The A-1 and A-2 compounds (A-1: 1.4 mg, 13%; A-2: 1.8 mg, 17%) were obtained by lyophilization as white solids; EI-MS m / z: A-1: ​​1357 [M+H] + ,A-2:1328[M+H] + .

[0713] Example I-2: Preparation of Compound A-3

[0714]

[0715] Step 1: Preparation of compound A-3a

[0716] Triphenylphosphine (65.6 mg, 0.24 mmol) and carbon tetrabromide (CBr4, 173 mg, 0.50 mmol) were dissolved in MC (5 mL) at 0 ° C under a nitrogen atmosphere, and the mixture was stirred for 30 minutes. Compound A-1d (160 mg, 0.20 mmol) prepared in step 4 of Example I-1 dissolved in MC (2 mL) was added to the above reaction solution, and the mixture was stirred for 1 hour. After the reaction was completed, EA (100 mL) and distilled water (100 mL) were added to extract the organic layer, and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to give compound A-3a (30 mg, 17%); EI-MS m / z: 761 [M+H] + .

[0717] Step 2: Preparation of compound A-3b

[0718] Compound A-3a (30 mg, 0.036 mmol) was dissolved in DMF (2 mL) at 0°C under a nitrogen atmosphere, and then auristatin F-OMe (30 mg, 0.04 mmol) and DIPEA (18.9 μL, 0.11 mmol) prepared in Preparation Example 9 were added in sequence, and the mixture was stirred at 60°C for 3 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the reaction solution containing compound A-3b was used for the next reaction without additional treatment; EI-MS m / z: 1509 [M+H] + .

[0719] Step 3: Preparation of compound A-3

[0720] Lithium hydroxide monohydrate (15.2 mg, 0.36 mmol) was slowly added dropwise to the reaction solution, and the mixture was stirred for 3 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (0.1 mL) was slowly added dropwise to terminate the reaction. The reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC. The product was lyophilized to obtain compound A-3 (7.9 mg, 16%) as a white solid; EI-MS m / z: 1299 [M+H] + .

[0721] Example I-3: Preparation of Compound A-4

[0722]

[0723] Step 1: Preparation of compound A-4a

[0724] Compound A-1b (300 mg, 0.69 mmol) prepared in step 2 of Example I-1 was dissolved in DMF (5 mL) at room temperature under a nitrogen atmosphere, and potassium carbonate (286 mg, 2.07 mmol) and compound L3-1 (424 mg, 0.90 mmol) prepared in Preparation Example 10 were added in sequence, and the mixture was stirred at 70 ° C for 4 hours. After the reaction was completed, the temperature of the reaction solution was lowered to room temperature, EA (100 mL) and distilled water (200 mL) were added, and the organic layer was extracted. Distilled water (300 mL) was added and the obtained organic layer was extracted again. The extracted organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-4a (580 mg, 96%) in the form of a light yellow oil.

[0725] 1H-NMR (400MHz, CDCl3) δ10.05(s,1H),8.04(s,1H),7.57(d,J=8Hz,1H),7.43(d,J=8.8Hz,2H),7.05(d,J= 8.8Hz,2H),6.88(d,J=8Hz,1H),5.52(m.2H),5.25(s,2H),5.15-5.08(m,4H),4.36(q,J=7.2Hz,2H),4.26- 4.22(m,1H),4.19-4.07(m,2H),3.69(t,J=6Hz,2H),3.59(m,2H),3.53-3.51(m,2H),3.48-3.43(m,4H),3 .41-3.39(m,2H),3.44(m,2H),2.19(s,3H),2.08(s,3H),2.05(s,3H),2.02(s,3H),1.41(t,J=7.2Hz,3H).

[0726] Step 2: Preparation of compound A-4b

[0727] Compound A-4a (580 mg, 0.66 mmol) was dissolved in THF (10 mL) at 0°C under a nitrogen atmosphere, and sodium borohydride (63 mg, 1.65 mmol) was added, and the mixture was stirred for 3 hours. After the reaction was completed, distilled water (100 mL) was added to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-4b (370 mg, 63%).

[0728] 1 H-NMR (400MHz, CDCl3) δ7.42-7.40(m,3H),7.04-6.98(m,3H),6.73(d,J=8Hz,1H), 5.52-5.47(m,2H),5.15-5.07(m,6H),4.89(m,2H),4.35(q,J=7.2Hz,2H),4.23(m, 1H),4.17(m,1H),4.09(m,1H),3.70(m,2H),3.59(m,2H),3.53-3.41(m,8H),3.33( m,2H),2.19(s,3H),2.08(s,3H),2.05(s,3H),2.02(s,3H),1.40(t,J=7.2Hz,3H).

[0729] Step 3: Preparation of compound A-4c

[0730] Compound A-4b (150 mg, 0.17 mmol) was dissolved in DMF (3 mL) at 0 ° C under a nitrogen atmosphere, and then bis (4-nitrophenyl) carbonate (104.5 mg, 0.34 mmol) and DIPEA (60 μL, 0.34 mmol) were added in sequence, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the organic layer was diluted with EA (50 mL), and distilled water (50 mL) and 2N-hydrochloric acid aqueous solution (10 mL) were added to extract the organic layer. The extracted organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-4c (115 mg, 67%).

[0731] 1 H-NMR (400MHz, CDCl3) δ8.20(d,J=8Hz,2H),7.41(d,J=8.8Hz,2H),7.29(s,1H),7.07-7.02(m,5H ),6.75(d,J=8Hz,1H),5.52-5.46(m,2H),5.34(s,2H),5.16(s,2H),5.14-5.07(m,4H),4.35(q,J =7.2Hz,2H),4.23(m,1H),4.19-4.08(m,2H),3.72(m,2H),3.61(t,J=7.2Hz,2H),3.52-3.42(m,8 EI-MS m / z:1016[M+Na] + .

[0732] Step 4: Preparation of compound A-4

[0733] Compound A-4c (22.3 mg, 0.022 mmol) was dissolved in methanol (1 mL) and THF (1 mL) at 0°C, and then lithium hydroxide monohydrate (4.7 mg, 0.112 mmol) dissolved in distilled water (200 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 10 minutes. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-4 (8.9 mg, 48%) as an ivory solid.

[0734] 1H-NMR (400MHz, DMSO-d6) δ8.24(d,J=9.2Hz,2H),7.49(d,J=8.8Hz,2H),7.34(s,1H),7.28( d,J=9.2Hz,2H),7.20(d,J=7.6Hz,1H),7.08(d,J=8.8Hz,2H),6.99(d,J=7.6Hz,1H),5.48( s,2H),5.21(s,2H),5.18(m,1H),5.03-5.01(m,2H),4.89-4.84(m,2H),4.67(m,1H),4.53( EI-MS m / z:848[M+Na] + .

[0735] Example I-4: Preparation of Compound A-5

[0736]

[0737] The compound A-4c (23.5 mg, 0.024 mmol) prepared in step 3 of Example I-3 was dissolved in methanol (1.5 mL) and THF (0.4 mL) at 0 ° C, and then lithium hydroxide monohydrate (24.8 mg, 0.59 mmol) dissolved in distilled water (500 μL) was slowly added dropwise, and the mixture was stirred at 0 ° C for 1 hour. The reaction mixture was stirred for another 2 hours at room temperature. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound A-5 (10 mg, 53%) as a white solid.

[0738] 1 H-NMR (400MHz, DMSO-d6) δ8.21(d,J=9.2Hz,2H),7.47(d,J=8.4Hz,2H),7.26( d,J=9.2Hz,2H),7.18(s,1H),7.12(d,J=8Hz,1H),7.06(d,J=8.4Hz,2H),6.91( d,J=8Hz,1H),5.43(s,2H),5.19(s,2H),5.17(m,1H),5.03-5.01(m,2H),4.90- 4.83(m,2H),4.65(m,1H),4.51(m,1H),3.71(m,1H),3.61-3.46(m,15H); EI-MS m / z:820[M+Na]+ .

[0739] Example I-5: Preparation of Compound A-6

[0740]

[0741] Step 1: Preparation of compound A-6a

[0742] Compound A-4b (50 mg, 0.057 mmol) prepared in step 2 of Example I-3 was dissolved in MC (2.5 mL) at 0°C under a nitrogen atmosphere, and then thionyl chloride (5 μL, 0.069 mmol) was added, and the mixture was stirred at 0°C for 3 hours. After the reaction was completed, EA (50 mL) and distilled water (50 mL) were added to extract the organic layer, which was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound A-6a (50 mg, 98%); EI-MS m / z: 891 [M+H] + .

[0743] Step 2: Preparation of compound A-6b

[0744] Compound A-6a (50 mg, 0.056 mmol) was dissolved in DMF (2 mL) at 0°C under a nitrogen atmosphere, and then auristatin F-OMe (46.9 mg, 0.061 mmol), DIPEA (24.4 μL, 0.14 mmol) and TBAI (2 mg, 0.005 mmol) prepared in Preparation Example 9 were added in sequence. The mixture was stirred at 40°C for 15 hours to obtain compound A-6b; EI-MS m / z: 1615 [M+H] + .

[0745] Step 3: Preparation of compound A-6

[0746] The reaction solution containing compound A-6b obtained in step 2 above was cooled to room temperature, and methanol (1 mL) was added. 6N-sodium hydroxide aqueous solution (0.1 mL) and distilled water (1 mL) were added sequentially at 0°C, and the mixture was stirred for 30 minutes, and then stirred at room temperature for another 2 hours. 2N-hydrochloric acid aqueous solution (0.1 mL) was slowly added dropwise to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC. After lyophilization, compound A-6 (2 mg, 2%) was obtained; EI-MS m / z: 1405 [M+H] + .

[0747] Example I-6: Preparation of Compound A-7

[0748]

[0749] Step 1: Preparation of compound A-7a

[0750] Core C-1 (99.1 mg, 0.42 mmol) was dissolved in MC (15 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (192 mg, 0.46 mmol) and benzyltributylammonium chloride (132 mg, 0.42 mmol) were added. 5N-sodium hydroxide aqueous solution (255 μL, 1.27 mmol) was slowly added to the reaction solution, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, MC (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-7a (131.3 mg, 55%) in the form of an ivory solid.

[0751] 1 H-NMR (400MHz, CDCl3) δ10.19(s,1H),7.82(d,J=8Hz,1H),7.38(s,1H),6.80(d,J=8Hz,1H),5.64(dd,J=10.4,8Hz,1H),5.50(d,J=3.2Hz,1H),5. 29(d,J=8Hz,1H),5.16(dd,J=10.4,3.2Hz,1H),4.34(s,3H),4.26-4.09( m,3H),3.93(s,3H),2.21(s,3H),2.08(s,3H),2.06(s,3H),2.05(s,3H).

[0752] Step 7: Preparation of compound A-7b

[0753] Compound A-7a (131.3 mg, 0.23 mmol) was dissolved in THF (3 mL) at 0 ° C under a nitrogen atmosphere, and then sodium borohydride (26.5 mg, 0.70 mmol) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, distilled water (50 mL) was added to terminate the reaction, and EA (50 mL) was added to extract the organic layer twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-7b (101.5 mg, 77%) as a yellow solid.

[0754] 1H-NMR (400MHz, CDCl3) δ7.32(s,1H),7.12(d,J=8Hz,1H),6.63(d,J=8Hz,1H),5.61(dd,J=10.4,8Hz,1H),5.48(d,J=2.8Hz,1H),5.1 6-5.12(m,2H),4.99-4.97(m,2H),4.41(s,3H),4.28-4.08(m,3H),3.91(s,3H),2.20(s,3H),2.07(s,3H),2.06(s,3H),2.04(s,3H).

[0755] Step 8: Preparation of compound A-7c

[0756] Compound A-7b (101.5 mg, 0.18 mmol) was dissolved in MC (5 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (108.5 mg, 0.54 mmol) and DIPEA (156 μL, 0.89 mmol) were added in sequence, and the mixture was stirred for 20 hours. After the reaction was completed, EA (30 mL) and distilled water (30 mL) were added, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-7c (44.1 mg, 36%).

[0757] 1 H-NMR (400MHz, CDCl3) δ8.25(d,J=9.2Hz,2H),7.35(s,1H),7.23(d,J=8Hz,1H),7.0 8(d,J=9.2Hz,2H),6.69(d,J=8Hz,1H),5.62(dd,J=10.4,8Hz,1H),5.49(d,J=2.8Hz ,1H),5.44(m,2H),5.18(d,J=8Hz,1H),5.15(dd,J=10.4,3.6Hz,1H),4.27(m,1H),4 .24(s,3H),4.20-4.09(m,2H),3.91(s,3H),2.21(s,3H),2.06(m,6H),2.04(s,3H).

[0758] Step 9: Preparation of compound A-7

[0759] Compound A-7c (37.2 mg, 0.054 mmol) was dissolved in methanol (1.5 mL) and ACN (1.5 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (52.4 mg, 0.38 mmol) was added, and the mixture was stirred for 1.5 hours while the temperature was raised from 0°C to room temperature. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound A-7 (5 mg, 18%) as a white solid.

[0760] 1 H-NMR (400MHz, DMSO-d6) δ8.25(d,J=9.2Hz,2H),7.47(s,1H),7.38(d,J=8Hz,1H),7.32(d,J=9.2Hz,2H),6.76(d,J=8Hz,1H) ,5.59(s,2H),5.33(m,1H),4.94-4.92(m,2H),4.68(m,1H),4.57(m,1H),4.18(s,3H),3.85(s,3H),3.73(m,1H),3.64(m,1H).

[0761] Example I-7: Preparation of Compound A-8

[0762]

[0763] Step 1: Preparation of compound A-8a

[0764] Core C-2 (2.4 g, 7.75 mmol) was dissolved in DMF (25 mL) at 0°C under a nitrogen atmosphere. EDC (CAS No. 25952-53-8, 2.23 g, 11.63 mmol), HOBt (1.05 g, 8.14 mmol), DMAP (95 mg, 0.77 mmol), DIPEA (4 mL, 23.25 mmol), and linker P-1 (1.98 g, 7.75 mmol) were then added in sequence, and the mixture was stirred for 30 minutes. The reaction mixture was stirred at room temperature for an additional 16 hours, then extracted with EA (300 mL) and 1N aqueous hydrochloric acid solution (400 mL). The resulting organic layer was extracted with 1N aqueous sodium hydroxide solution (5 mL) and distilled water (300 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to yield compound A-8a (2.57 g, 65%) as a pale yellow oil.

[0765] 1H-NMR (400MHz, CDCl3) δ10.01 (s, 1H), 7.67 (s, 1H), 7.54 (d, J = 8Hz, 1H), 7.48-7.37 (m, 5H), 6 .88-6.84(m,2H),5.30(s,2H),4.33(s,3H),3.71-3.60(m,14H),3.32(t,J=5.2Hz,2H); EI-MS m / z:510[M+H] + .

[0766] Step 2 to Step 7: Preparation of Compound A-8

[0767] Using compound A-8a as a starting material, compound A-8 (3.1 mg, 39%) was obtained as a white solid in the same manner as steps 2 to 7 of Example I-1; EI-MS m / z: 1341 [M+H] + .

[0768] Example I-8: Preparation of Compound A-9

[0769]

[0770] Using compound A-8b as a starting material, compound A-9 (21.5 mg, 72%) was obtained as an ivory solid in the same manner as in Example I-3.

[0771] 1 H-NMR(400MHz,DMSO-d6)δ8.50(t,J=5.2Hz,1H),8.22(d,J=9.2Hz,2H),7.45(d, J=8.8Hz,2H),7.26(d,J=9.2Hz,2H),7.13-7.11(m,2H),7.06(d,J=8.8Hz,2H),6. 87(d,J=8Hz,1H),5.38(s,2H),5.18(s,2H),5.16(m,1H),4.84-4.82(m,2H),4.64 (m,1H),4.51(m,1H),4.20(s,3H),3.70(m,1H),3.56-3.51(m,15H),3.40(m,2H).

[0772] Example I-9: Preparation of Compound A-10

[0773]

[0774] Step 1: Preparation of compound A-10a

[0775] Core C-3 (160 mg, 0.89 mmol) was dissolved in ACN (12 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (406 mg, 0.98 mmol), silver (I) oxide (520 mg, 2.22 mmol) and molecular sieves (70 mg) were added in sequence, and the mixture was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction solution was diluted with EA (100 mL), filtered using a diatomaceous earth filter, and extracted by adding distilled water (100 mL) to the solution. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-10a (400 mg, 87.7%) as a solid.

[0776] 1 H-NMR (400MHz, CDCl3) δ10.13(s,1H),7.84(d,J=8Hz,1H),7.59(d,J=5.6Hz,1H),7.49(d,J=5.6Hz,1H),7.09(d,J=8Hz,1H),5.67(dd,J=10.4,8Hz,1 H),5.52(d,J=3.2Hz,1H),5.30(d,J=8Hz,1H),5.19(dd,J=10.4,3.6Hz,1H ),4.29(m,1H),4.21-4.17(m,2H),2.21(s,3H),2.10(s,3H),2.04(s,6H).

[0777] Step 2: Preparation of compound A-10b

[0778] Compound A-10a (183 mg, 0.36 mmol) was dissolved in THF (10 mL) at 0 ° C. under a nitrogen atmosphere, and then sodium borohydride (34 mg, 0.90 mmol) was added, and the mixture was stirred at 0 ° C. for 1 hour, and then the mixture was stirred at room temperature for 2.5 hours. After the reaction was completed, distilled water (50 mL) was added to terminate the reaction, and EA (50 mL) was added to extract the organic layer twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-10b (137.2 mg, 75%) as a white solid.

[0779] 1H-NMR (400MHz, CDCl3) δ7.45(d,J=5.6Hz,1H),7.40(d,J=5.6Hz,1H),7.28(d,J=8Hz,1H),6.95(d,J=8Hz,1H),5.63(dd,J=10.4,8Hz,1H),5.50(d,J =2.8Hz,1H),5.17-5.14(m,2H),4.92(d,J=5.6Hz,2H),4.29-4.11(m,3H) ,2.20(s,3H),2.08(s,3H),2.05(s,3H),2.03(s,3H),1.79(t,J=6Hz,1H).

[0780] Step 3: Preparation of compound A-10c

[0781] Compound A-10b (137.2 mg, 0.18 mmol) was dissolved in MC (10 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (81.2 mg, 0.40 mmol) and DIPEA (140 μL, 0.81 mmol) were added in sequence, and the mixture was stirred for 16 hours while slowly raising the temperature from 0 ° C to room temperature. After the reaction was completed, EA (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-10c (114.7 mg, 63%) as a white solid.

[0782] 1 H-NMR (400MHz, CDCl3) δ8.27(d,J=9.2Hz,2H),7.48(d,J=5.6Hz,1H),7.44(d,J=5.6Hz,1H),7.39-7.36(m,3H),6.97(d,J=8Hz,1H),5.64(dd,J =10.4,8Hz,1H),5.24(s,2H),5.51(d,J=3.2Hz,1H),5.19-5.14(m,2H), 4.29-4.13(m,3H),2.21(s,3H),2.08(s,3H),2.05(s,3H),2.04(s,3H).

[0783] Step 4: Preparation of compound A-10d

[0784] Compound A-10c (64 mg, 0.094 mmol) was dissolved in DMF (3 mL) at 0 ° C under a nitrogen atmosphere, and then compound PL-1 (25 mg, 0.113 mmol), HOBt (20 mg, 0.141 mmol), pyridine (500 μL) and DIPEA (41 μL, 0.235 mmol) were added in sequence, and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, EA (50 mL) and 1N-hydrochloric acid aqueous solution (50 mL) were used for extraction. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-10d (49 mg, 68.3%).

[0785] 1 H-NMR (400MHz, CDCl3) δ7.46-7.39(m,2H),7.34(m,1H),7.28-7.22(m,2H),7.16 -7.03(m,3H),7.01-6.65(m,2H),5.63(m,1H),5.50(m,1H),5.17-4.93(m,4H),4 .63-4.09(m,4H),3.26-3.22(m,3H),2.88-3.87(m,3H),2.32(m,1H),2.20(s,3H ),2.08-2.07(m,3H),2.05-2.03(m,6H),0.90-0.87(m,3H),0.72-0.55(m,3H); MS m / z:757[M+H] + .

[0786] Step 5: Preparation of Compound A-10

[0787] Compound A-10d (49 mg, 0.065 mmol) was dissolved in methanol (2 mL) at 0°C, and lithium hydroxide monohydrate (13.6 mg, 0.324 mmol) dissolved in distilled water (400 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 10 minutes. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-10 (28.5 mg, 74%) as a white solid.

[0788] 1H-NMR(400MHz,DMSO-d6)δ7.72(m,0.5H),7.68-7.65(m,1H),7.58(m,0.5H),7.31-7.21(m,3H),7.15-7.02(m,4H),5.31(m,1H),5.10-4.91(m,3H) ),4.68(m,1H),4.55(m,1H),4.24(m,1H),3.73-3.44(m,3H),3.14(s,3H ),2.72(m,3H),2.16(m,1H),0.83-0.77(m,3H),0.63-0.43(m,3H); EI-MS m / z:589[M+H] + .

[0789] Example I-10: Preparation of Compound A-11

[0790]

[0791] In step 4 of Example I-9, except that MMAF-OMe was used instead of compound PL-1, compound A-11 (10.4 mg, 65%) was obtained in the same manner as in Example I-9 as a white solid; EI-MS m / z: 1101 [M+H] + .

[0792] Example I-11: Preparation of Compound A-12

[0793]

[0794] In addition to using MMAF-OMe instead of compound PL-1 in step 4 of Example I-9, core C-5 was used as a starting material to obtain compound A-12 (11.7 mg, 64%) as a white solid in the same manner as in Example I-9; EI-MS m / z: 1145 [M+H] + .

[0795] Example I-12: Preparation of Compound A-13

[0796]

[0797] Step 1: Preparation of compound A-13a

[0798] Core C-6 (192 mg, 0.61 mmol) was dissolved in DMF (5 mL) at 0°C under a nitrogen atmosphere, followed by the addition of EDCI (177 mg, 0.92 mmol), HOBt (83.1 mg, 0.61 mmol), DMAP (7.5 mg, 0.06 mmol), and DIPEA (322 μL, 1.85 mmol). Linker P-4 (azido-PEG4-amine, TCI, CAS No. 951671-92-4, 184 mg, 0.61 mmol) dissolved in DMF (3 mL) was added, and the mixture was stirred at 0°C for 3 hours. The mixture was stirred for another 16 hours while the temperature was raised to room temperature. After the reaction was complete, the organic layer was extracted with EA (100 mL) and 2N-hydrochloric acid aqueous solution (100 mL). 2N-sodium hydroxide aqueous solution (5 mL) and distilled water (50 mL) were added to the resulting organic layer, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-13a (184.9 mg, 54%) as a brown oil.

[0799] 1 H-NMR (400MHz, CDCl3) δ10.07 (s, 1H), 8.12 (s, 1H), 7.89 (d, J = 8.4Hz, 1H), 7.50-7.37 (m, 5H), 7 .00(d,J=8Hz,1H),6.96(m,1H),5.33(s,2H),3.68-3.58(m,18H),3.31(t,J=5.2Hz,2H); EI-MS m / z:557[M+H] + .

[0800] Step 2: Preparation of compound A-13b

[0801] Compound A-13a (184.9 mg, 0.33 mmol) was dissolved in MC (10 mL) at -50 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 2.99 mL, 2.99 mmol) was slowly added, and the mixture was stirred for 3.5 hours while slowly warming to -30 ° C. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (100 mL) was added to the mixture and the organic layer was extracted twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue obtained by column chromatography gave compound A-13b (75.9 mg, 49%) in the form of a light orange solid.

[0802] 1H-NMR (400MHz, CDCl3) δ10.02 (s, 1H), 8.41 (s, 1H), 7.84 (d, J = 8Hz, 1H), 7.32 (brs, 1H) ,6.98(d,J=8Hz,1H),3.82-3.64(m,16H),3.55(t,J=5.2Hz,2H),3.28(t,J=5.2Hz,2H).

[0803] Steps 3 to 7: Preparation of Compound A-13

[0804] In step 4 of Example I-9, except that MMAF-OMe was used instead of compound PL-1, compound A-13 (25.7 mg, 66%) was obtained in the same manner as in Example I-9 as a white solid; EI-MS m / z: 1389 [M+H] + .

[0805] Example I-13: Preparation of Compound A-14

[0806]

[0807] In step 1 of Example I-12, except that linker P-5 was used instead of linker P-4, compound A-14 (22.4 mg, 67%) was obtained in the same manner as Example I-12 as a white solid; EI-MS m / z: 1358 [M+H] + .

[0808] Example I-14: Preparation of Compound A-15

[0809]

[0810] Step 1: Preparation of compound A-15a

[0811] Core C-7 (278 mg, 0.81 mmol) and linker P-1 (208 mg, 0.81 mmol) were dissolved in DMF (5 mL) at 0 ° C under a nitrogen atmosphere, and then EDCI hydrochloride (235 mg, 1.22 mmol), DMAP (10 mg, 0.08 mmol) and DIPEA (430 μL, 2.45 mmol) were added in sequence, and the mixture was stirred for 16 hours while slowly warming to room temperature. After the reaction was completed, the organic layer was extracted twice with EA (250 mL) and 2N-hydrochloric acid aqueous solution (250 mL). Saturated sodium chloride aqueous solution (250 mL) was added to the obtained organic layer, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-15a (207 mg, 47%) in the form of a yellow oil.

[0812] 1 H-NMR (400MHz, CDCl3) δ10.03(s,1H),7.74(d,J=8Hz,1H),7.49-7.34(m,6H),6.94(d,J=8Hz,1H),6.18(m,1H),5.30(s,2H),3.65-3.60(m, 6H),3.57-3.55(m,2H),3.52-3.48(m,4H),3.44(t,J=5.2Hz,2H),3.35(t,J=5.2Hz,2H),3.31(t,J=7.6Hz,2H),2.64(t,J=7.6Hz,2H); EI-MS m / z:541[M+H] + .

[0813] Step 2: Preparation of compound A-15b

[0814] Compound A-15a (207 mg, 0.38 mmol) was dissolved in MC (10 mL) at -55 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 2.3 mL, 2.3 mmol) was slowly added, and the mixture was stirred for 2.5 hours while slowly raising the temperature to -40 ° C. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (100 mL) was added to the mixture and the organic layer was extracted twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-15b (97.5 mg, 56%) in the form of a yellow viscous glue.

[0815] 1 H-NMR (400MHz, CDCl3) δ9.98 (s, 1H), 7.66 (d, J = 8Hz, 1H), 7.44 (s, 1H), 6.89 (d, J = 8Hz, 1H), 6.36 (brs, 1H), 3.66 ( EI-MS m / z:451[M+H] + .

[0816] Step 3: Preparation of compound A-15c

[0817] Compound A-15b (97.5 mg, 0.21 mmol) was dissolved in MC (7 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (142 mg, 0.34 mmol) and benzyltributylammonium chloride (67.5 mg, 0.21 mmol) were added. 5N-sodium hydroxide aqueous solution (0.13 mL, 0.65 mmol) was slowly added to the reaction solution, and the mixture was stirred for 20 hours while the temperature was slowly raised to room temperature. After the reaction was completed, MC (150 mL) and distilled water (150 mL) were added, and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-15c (42.3 mg, 25%).

[0818] 1 H-NMR (400MHz, CDCl3) δ10.08(s,1H),7.75(d,J=8.4Hz,1H),7.18(s,1H),6.05(d,J=8.4Hz,1H),6.14(m,1H), 5.59(dd,J=10.4,8Hz,1H),5.52(d,J=3.2Hz,1H),5.27(d,J=8Hz,1H),5.18(dd,J=10.4,3.2Hz,1H),4.27(m,1 H),4.20-4.15(m,2H),3.67-3.65(m,6H),3.61-3.59(m,2H),3.57-3.52(m,4H),3.48-3.45(m,2H),3.37(t,J= 5.2Hz,2H),3.16(t,J=7.6Hz,2H),2.63(t,J=7.6Hz,2H),2.21(s,3H),2.09(s,3H),2.06(s,3H),2.04(s,3H).

[0819] Step 4: Preparation of compound A-15d

[0820] Compound A-15c (42.3 mg, 0.054 mmol) was dissolved in THF (3 mL) at 0°C under a nitrogen atmosphere. Sodium borohydride (5.1 mg, 0.135 mmol) was then added, and the mixture was stirred for 2.5 hours while slowly raising the temperature from 0°C to room temperature. After completion, the reaction was terminated by adding distilled water (50 mL), and the organic layer was extracted twice with EA (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was purified by column chromatography to yield compound A-15d (30.6 mg, 72%) as a white solid; EI-MS m / z: 783 [M+H]+.

[0821] Step 5: Preparation of compound A-15e

[0822] Compound A-15d (30.6 mg, 0.039 mmol) was dissolved in MC (2 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (11.8 mg, 0.058 mmol) and DIPEA (17 μL, 0.098 mmol) were added in sequence, and the mixture was stirred for 16 hours while the temperature was raised from 0 ° C to room temperature. After the reaction was completed, MC (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-15e (22.2 mg, 60%) as a white solid; EI-MS m / z: 948 [M + H] + .

[0823] Step 6: Preparation of Compound A-15f

[0824] Compound A-15e (22.2 mg, 0.023 mmol) was dissolved in DMF (0.5 mL) at 0 ° C. under a nitrogen atmosphere, and then MMAF-OMe (17.5 mg, 0.023 mmol), HOBt (4.7 mg, 0.035 mmol), pyridine (500 μL), DIPEA (10.2 μL, 0.058 mmol) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted twice with EA (50 mL) and 2N-hydrochloric acid aqueous solution (50 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-15f (23.6 mg, 65%) as a colorless solid; EI-MS m / z: 1555 [M+H] + .

[0825] Step 7: Preparation of Compound A-15

[0826] Compound A-15f (23.6 mg, 0.015 mmol) was dissolved in methanol (0.6 mL) and THF (0.3 mL) at 0°C, and then lithium hydroxide monohydrate (6.3 mg, 0.152 mmol) dissolved in distilled water (0.2 mL) was slowly added dropwise, and the mixture was stirred for 2.5 hours while the temperature was slowly raised from 0°C to room temperature. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and then lyophilized to obtain compound A-15 (10.1 mg, 48%) as a white solid; EI-MS m / z: 1373 [M+H] + .

[0827] Example I-15: Preparation of Compound A-16

[0828]

[0829] In addition to using MMAF-OMe instead of compound PL-1 in step 4 of Example I-9, core C-8 was used as a starting material to obtain compound A-16 (7.7 mg, 60%) as a white solid in the same manner as in Example I-9; EI-MS m / z: 1172 [M+H] + .

[0830] Example I-16: Preparation of Compound A-17

[0831]

[0832] Step 1: Preparation of compound A-17a

[0833] Core C-4 (174 mg, 0.976 mmol) was dissolved in MC (30 mL) at room temperature under a nitrogen atmosphere, followed by the addition of acetyl bromide-α-D-galactose (440 mg, 1.07 mmol) and benzyltributylammonium chloride (Sigma-Aldrich, CAS No.: 23616-79-7, 304 mg, 0.976 mmol). 5N-sodium hydroxide aqueous solution (586 μL, 2.93 mmol) was added to the reaction solution, and the mixture was stirred for 5 hours. After the reaction was completed, MC (100 mL) and distilled water (100 mL) were added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound A-17a (250 mg, 50.4%).

[0834] 1 H-NMR (400MHz, CDCl3) δ10.09(s,1H),8.09(s,1H),7.59(d,J=8Hz,1H),7.44(t,J=8Hz,1H),6.95(d,J=8Hz,1H),5.64(dd,J=10.4,8Hz,1H),5 .50(d,J=3.2Hz,1H),5.23(d,J=8Hz,1H),5.18(dd,J=10.4,8Hz,1H),4.27(m,1H),4.20-4.11(m,2H),2.21(s,3H),2.06(s,6H),2.05(s,3H).

[0835] Step 2: Preparation of compound A-17b

[0836] Compound A-17a (250 mg, 0.49 mmol) was dissolved in THF (10 mL) at 0°C under a nitrogen atmosphere, and sodium borohydride (46.5 mg, 1.225 mmol) was added, and the mixture was stirred for 1 hour. After the reaction was completed, distilled water (50 mL) was added to terminate the reaction, and EA (50 mL) was added to extract the organic layer. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-17b (230 mg, 92%).

[0837] 1 H-NMR (400MHz, CDCl3) δ7.53(d,J=8Hz,1H),7.28-7.22(m,2H),6.93(d,J=8Hz,1H),5.60(dd,J=10.4,8Hz,1H),5.48(d,J=2.4Hz,1H),5.1 6-5.12(m,2H),4.91(d,=5.2Hz,2H),4.31(dd,J=11.2,7.2Hz,1H),4.16-4.10(m,2H),2.21(s,3H),2.07(s,3H),2.04(s,3H),2.03(s,3H).

[0838] Step 3: Preparation of compound A-17c

[0839] Compound A-17b (300 mg, 0.58 mmol) was dissolved in concentrated hydrochloric acid (8 mL) at 0°C under a nitrogen atmosphere, and the mixture was stirred at 0°C for 1 hour. After the reaction was completed, EA (50 mL) and distilled water (50 mL) were added to extract the organic layer, and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain compound A-17c (300 mg, 96.7%) as a white solid; MS m / z: 529 [M+H] + .

[0840] Step 4: Preparation of compound A-17d

[0841] Compound A-17c (300 mg, 0.56 mmol) was dissolved in DMF (15 mL) at room temperature under a nitrogen atmosphere, followed by the addition of sodium azide (55 mg, 0.84 mmol), and the mixture was stirred at 60°C for 2 hours. After the reaction was complete, EA (100 mL) and distilled water (200 mL) were added to extract the organic layer, which was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound A-17d (230 mg, 75.6%).

[0842] 1H-NMR (400MHz, CDCl3) δ7.52(d,J=8Hz,1H),7.32(s,1H),7.28(t,J=8Hz,1H),6 .94(d,J=8Hz,1H),5.61(dd,J=10.4,8Hz,1H),5.50(d,J=2.4Hz,1H),5.17-5.14 (m,2H),4.60(d,J=14.4Hz,1H),4.52(d,J=14.4Hz,1H),4.26(dd,J=11.2,6.8Hz ,1H),4.20-4.11(m,2H),2.20(s,3H),2.07(s,3H),2.05(s,3H),2.04(s,3H); MS m / z:558[M+Na] + .

[0843] Step 5: Preparation of compound A-17e

[0844] A-17d (230 mg, 0.42 mmol) was dissolved in MC (15 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (116 μL, 1.26 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 1.28 mL, 1.26 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 1 hour. After the reaction was completed, cooled distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-17e (130 mg, 53.7%) as a white solid.

[0845] 1 H-NMR (400MHz, CDCl3) δ10.11(s,1H),7.84(d,J=8Hz,1H),7.40(s,1H),7.08(d,J=8Hz,1H),5.66(dd,J=10.4,8Hz,1H),5.52(d,J=3.2Hz,1H),5.29(d, J=8Hz,1H),5.19(dd,J=10.4,3.2Hz,1H),4.65(d,J=14.4Hz,1H),4.52(d,J =14.4Hz,1H),4.31-4.17(m,3H),2.21(s,3H),2.09(s,3H),2.05(s,6H);MS m / z:586[M+Na] + .

[0846] Step 6: Preparation of Compound A-17f

[0847] Compound A-17e (130 mg, 0.23 mmol) was dissolved in THF (10 mL) at 0°C under a nitrogen atmosphere, followed by the addition of sodium borohydride (22 mg, 0.575 mmol), and the mixture was stirred for 1 hour. After completion of the reaction, distilled water (100 mL) was added to terminate the reaction, and the organic layer was extracted with EA (100 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound A-17f (120 mg, 92.3%).

[0848] 1 H-NMR (400MHz, CDCl3) δ7.36 (s, 1H), 7.28 (d, J = 8Hz, 1H), 6.94 (d, J = 8Hz, 1H), 5.61 (d d,J=10.4,8Hz,1H),5.49(d,J=2.8Hz,1H),5.17-5.14(m,2H),4.89(d,J=5.6Hz,2H),4 .61(d,J=14.4Hz,1H),4.54(d,J=14.4Hz,1H),4.26(dd,J=11.2,6.8Hz,1H),4.20-4. 11(m,2H),2.20(s,3H),2.07(s,3H),2.05(s,3H),2.04(s,3H),1.79(t,J=5.6Hz,1H).

[0849] Step 7: Preparation of Compound A-17g

[0850] Compound A-17f (120 mg, 0.21 mmol) was dissolved in MC (3 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (85 mg, 0.42 mmol), pyridine (51 μL, 0.63 mmol) and DIPEA (55 μL, 0.32 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 30 minutes and at room temperature for 2 hours. After the reaction was completed, the reaction solution was diluted with MC (50 mL), 2N-hydrochloric acid aqueous solution (50 mL) was added, and the organic layer was extracted twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-17g (116.3 mg, 75%).

[0851] 1H-NMR (400MHz, CDCl3) δ8.27(d,J=9.2Hz,2H),7.39-7.37(m,4H),6.96(d,J=8Hz,1H),5.62(dd,J=10.4,8Hz,1H),5.50(d,J=3.2Hz,1H),5.48(s,2H),5 .19-5.15(m,2H),4.63(d,J=14.4Hz,1H),4.57(d,J=14.4Hz,1H),4.26(m,1 H),4.20-4.11(m,2H),2.21(s,3H),2.07(s,3H),2.05(s,3H),2.04(s,3H).

[0852] Step 8: Preparation of compound A-17h

[0853] Compound A-17g (18.7 mg, 0.025 mmol) and compound PL-1 (6.7 mg, 0.031 mmol) were dissolved in DMF (500 μL) at 0 ° C. under a nitrogen atmosphere, and then HOBt (5.2 mg, 0.038 mmol), pyridine (500 μL) and DIPEA (11.1 μL, 0.064 mmol) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the organic layer was extracted twice with EA (50 mL) and 2N-hydrochloric acid aqueous solution (50 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-17h (17.9 mg, 86%) as a white solid.

[0854] 1 H-NMR(400MHz, CDCl3)δ7.37-7.32(m,1H),7.28-7.22(m,2H),7.19-6.85 (m,5H),5.61(m,1H),5.50(m,1H),5.18-5.09(m,4H),4.64-4.48(m,3H), 4.39-4.13(m,3H),3.25-3.23(m,3H),2.90-2.88(m,3H),2.32(m,1H),2. 20(s,3H),2.07-2.04(m,9H),0.98-0.83(m,3H),0.72-0.55(m,3H); EI-MS m / z:812[M+H] + .

[0855] Step 9: Preparation of Compound A-17

[0856] Compound A-17h (17.9 mg, 0.022 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (21.3 mg, 0.154 mmol) was added, and the mixture was stirred at 0°C for 40 minutes. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-17 (7.5 mg, 53%) as a white solid; EI-MS m / z: 644 [M+H] + .

[0857] Example I-17: Preparation of Compound A-18

[0858]

[0859] In step 8 of Example I-16, except that MMAF-OMe was used instead of compound PL-1 and lithium hydroxide monohydrate was used instead of potassium carbonate in step 9, compound A-18 (22 mg, 71%) was obtained in the same manner as in Example I-16 as a white solid; EI-MS m / z: 1156 [M+H] + .

[0860] Example I-18: Preparation of Compound A-19

[0861]

[0862] Compound A-18 (4.3 mg, 0.0037 mmol) was dissolved in DMSO (200 μL) at 0°C under a nitrogen atmosphere, and then 70 μL of a solution of linker P-6 (16.7 mg) dissolved in DMSO (1 mL) was added to the reaction. Distilled water (2 mL), DMSO (0.4 mL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (1.47 mg) were added to the reaction solution in sequence, and the mixture was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction solution was diluted with distilled water (1 mL), separated and purified using preparative HPLC, and then lyophilized to obtain compound A-19 (3.6 mg, 72%); MS m / z: 1358 [M+H] + .

[0863] Example I-19: Preparation of Compound A-20

[0864]

[0865] Compound A-18 (17.8 mg, 0.0154 mmol) was dissolved in THF (1 mL) at room temperature under a nitrogen atmosphere, and triphenylphosphine (4.4 mg, 0.017 mmol) and distilled water (0.3 mL) were added in sequence, and the mixture was stirred for 12 hours. 2N-aqueous sodium hydroxide solution (10 μL) was added to the reaction solution at room temperature, and the mixture was stirred for 30 minutes. 2N-aqueous hydrochloric acid solution was then slowly added dropwise to adjust the pH of the reaction solution to 3. The reaction solution was separated and purified by preparative HPLC, and then lyophilized to obtain compound A-20 (11.3 mg, 65%); MS m / z: 1130 [M+H] + .

[0866] Example I-20: Preparation of Compound A-21

[0867]

[0868] Compound A-20 (4.7 mg, 0.0041 mmol) was dissolved in THF (2 mL) at 0°C under a nitrogen atmosphere, and then linker P-7 (2.7 mg, 0.0078 mmol), DIPEA (10 μL) and distilled water (200 μL) were added dropwise in sequence, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was diluted with ACN (2 mL) containing 0.1% formic acid, separated and purified using preparative HPLC, and then lyophilized to obtain compound A-21 (2.9 mg, 51%); MS m / z: 1358 [M+H] + .

[0869] Example I-21: Preparation of Compound A-22

[0870]

[0871] Using core C-10 as a starting material, compound A-22 (8 mg, 58%) was obtained in the same manner as Example I-15 as a white solid; EI-MS m / z: 1171 [M+H] + .

[0872] Example I-22: Preparation of Compound A-23

[0873]

[0874] Step 1: Preparation of compound A-23a

[0875] Core C-11 (189 mg, 0.76 mmol) and linker P-1 (233 mg, 0.91 mmol) were dissolved in DMF (5 mL) at 0 ° C under a nitrogen atmosphere, and then EDCI hydrochloride (219 mg, 1.14 mmol), DMAP (9.3 mg, 0.07 mmol) and DIPEA (400 μL, 2.29 mmol) were added in sequence, and the mixture was stirred for 16 hours while slowly warming to room temperature. After the reaction was completed, the organic layer was extracted twice with EA (150 mL) and 2N-hydrochloric acid aqueous solution (150 mL). Saturated sodium chloride aqueous solution (100 mL) was added to the obtained organic layer, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-23a (47.5 mg, 14%) as an orange solid; EI-MSm / z: 449 [M+H] + .

[0876] Steps 2 to 6: Preparation of Compound A-23

[0877] In addition to using MMAF-OMe instead of compound PL-1 in step 4 of Example I-9, compound A-23a was used as a starting material, and the same method as Example I-9 was used to obtain compound A-23 (16.8 mg, 69%) as a white solid; EI-MS m / z: 1371 [M+H] + .

[0878] Example I-23: Preparation of Compound A-24

[0879]

[0880] Using core C-12 as a starting material, compound A-24 (12.1 mg, 68%) was obtained as a white solid in the same manner as steps 3 to 7 of Example I-1; EI-MS m / z: 1129 [M+H] + .

[0881] Example I-24: Preparation of Compound A-25

[0882]

[0883] Using core C-13 as a starting material, compound A-25 (18.6 mg, 65%) was obtained in the same manner as Example I-14 as a white solid; EI-MS m / z: 1342 [M+H] + .

[0884] Example I-25: Preparation of Compound A-26

[0885]

[0886] In addition to using MMAF-OMe instead of compound PL-1 in step 8 of Example I-16 and lithium hydroxide monohydrate instead of potassium carbonate in step 9, core C-14 was used as a starting material and the same method as in Example I-16 was used to obtain compound A-26 (21.3 mg, 61%) as a white solid; EI-MS m / z: 1140 [M+H] + .

[0887] Example I-26: Preparation of Compound A-27

[0888]

[0889] Compound A-26 (15.7 mg, 0.013 mmol) was dissolved in THF (1 mL) at room temperature under a nitrogen atmosphere, and triphenylphosphine (5.4 mg, 0.0195 mmol) and distilled water (0.3 mL) were added in sequence, and the mixture was stirred for 16 hours. 2N-aqueous sodium hydroxide solution (10 μL) was added to the reaction solution at room temperature, and stirred for another hour. 2N-aqueous hydrochloric acid solution was then slowly added dropwise to adjust the pH of the reaction solution to 3. The reaction solution was separated and purified using preparative HPLC, and then lyophilized to obtain compound A-27 (10.5 mg, 65%) as a white solid; MS m / z: 1113 [M+H] + .

[0890] Example I-27: Preparation of Compound A-28

[0891]

[0892] Compound A-27 (3 mg, 0.0026 mmol) was dissolved in THF (2 mL) at 0°C under a nitrogen atmosphere, and then linker P-7 (1.4 mg, 0.0039 mmol), DIPEA (10 μL) and distilled water (200 μL) were added dropwise in sequence, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, 2N-hydrochloric acid aqueous solution (20 μL) was added, and the reaction solution was diluted with ACN (2 mL) containing 0.1% formic acid, separated and purified using preparative HPLC, and then lyophilized to obtain compound A-28 (2.2 mg, 61%); MS m / z: 1341 [M+H] + .

[0893] Example I-28: Preparation of Compound A-29

[0894]

[0895] In step 2 of Example I-9, except that IPA and chloroform were used instead of THF as solvents, silica gel and sodium borohydride were added, and MMAF-OMe was used instead of compound PL-1 in step 4, core C-15 was used as a starting material, and the same method as in Example I-9 was used to obtain compound A-29 (15.9 mg, 64%) as a white solid; EI-MS m / z: 1192 [M+H] + .

[0896] Example I-29: Preparation of Compound A-33

[0897]

[0898] Step 1: Preparation of compound A-33a

[0899] The compound A-12b (180 mg, 0.31 mmol) of Example I-11 was dissolved in MC (20 mL) at 0 ° C under a nitrogen atmosphere, and then thionyl chloride (48.2 μ L, 0.65 mmol) was slowly added, and the mixture was stirred at 0 ° C for 2 hours. After the reaction was completed, MC (50 mL) was added to dilute the mixture, which was then concentrated under reduced pressure. The residue was solidified by adding n-hexane (50 mL), and then concentrated under reduced pressure to obtain compound A-33a in the form of a white solid. The obtained compound was used for the next step reaction (185 mg, 99%) without the need for additional purification.

[0900] 1 H-NMR (400MHz, CDCl3) δ8.12(s,1H),7.41(d,J=8Hz,1H),6.98(d,J=8Hz,1H),5.62(dd,J=10.8,8Hz,1H),5.48(d,J=2.8Hz,1H),5.17-5 .13(m,2H),4.80(s,2H),4.22(m,1H),4.18(m,1H),4.12(m,1H),3.96(s,3H),2.21(s,3H),2.08(s,3H),2.07(s,3H),2.04(s,3H); EI-MS m / z:587[M+H] + .

[0901] Steps 2 and 3: Preparation of Compound A-33

[0902] Compound A-33a (30 mg, 0.051 mmol) was dissolved in DMF (3 mL) at 0°C under a nitrogen atmosphere, and then compound auristatin F-OMe (42.7 mg, 0.056 mmol), DIPEA (26.7 μL, 0.153 mmol) and TBAI (2 mg, 0.005 mmol) were added in sequence, and the mixture was stirred at 40°C for 16 hours. After the reaction was completed, the reaction solution containing the generated compound A-33b (EI-MS m / z: 1311 [M+H]+) was cooled to room temperature, lithium hydroxide hydrate (21.4 mg, 0.51 mmol) was dissolved in distilled water (1 mL), slowly added at 0°C, and the mixture was stirred for 2 hours. 2N-hydrochloric acid aqueous solution (0.3 mL) was slowly added dropwise to the reaction solution to terminate the reaction. The reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and then lyophilized to obtain compound A-33 (1.1 mg, 2%); EI-MS m / z: 1115 [M+H] + .

[0903] Example I-30: Preparation of Compound A-34

[0904]

[0905] Step 1: Preparation of compound A-34a

[0906] Compound A-33a (135 mg, 0.229 mmol) of Example 1-29 was dissolved in ACN (5 mL) at room temperature under a nitrogen atmosphere, and potassium thioacetate (31.5 mg, 0.275 mmol) was added, and the mixture was stirred for 3 hours. After the reaction was completed, EA (50 mL) and distilled water (50 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-34a (140 mg, 97%).

[0907] 1 H-NMR (400MHz, CDCl3) δ8.11 (s, 1H), 7.40 (d, J = 8Hz, 1H), 6.95 (d, J = 8Hz, 1H), 5.60 (dd, J = 10.4, 8Hz, 1H), 5.47 (d, J = 2.8Hz, 1H), 5.15–5. 12(m,2H),4.31(d,J=5.2Hz,1H),4.25–4.08(m,3H),3.95(s,3H),2.36(s,3H),2.20(s,3H),2.08(s,2H),2.06(s,3H),2.03(s,3H); EI-MS m / z:627[M+H]+ .

[0908] Step 2: Preparation of compound A-34b

[0909] N-chlorosuccinimide (NCS, 120 mg, 0.88 mmol) was added to a mixed solution of 2N-hydrochloric acid aqueous solution (60 μL) and ACN (300 μL) at 0 ° C under a nitrogen atmosphere, and compound A-34a (140 mg, 0.22 mmol) was dissolved in ACN (100 μL) and added. The reaction solution was stirred at 0 ° C for 3 hours. After the reaction was completed, ether (100 mL) and distilled water (100 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain compound A-34b in the form of a white solid, which was used for the next reaction without additional purification (35 mg, 24%).

[0910] Steps 3 and 4: Preparation of Compound A-34

[0911] Compound A-34b (35 mg, 0.053 mmol) was dissolved in DMF (3 mL) at 0°C under a nitrogen atmosphere, and then SN-38 (CAS No.: 86639-52-3, 21 mg, 0.053 mmol) and TEA (18.7 μL, 0.132 mmol) were added sequentially, and the mixture was stirred at 0°C for 1 hour. After the reaction was completed, EA (50 mL) and 2N-hydrochloric acid aqueous solution (10 mL) and distilled water (100 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain compound A-34c. Methanol (2 mL) and THF (1 mL) were added to dissolve the obtained compound A-34c, and then lithium hydroxide hydrate (22.5 mg, 0.53 mmol) was dissolved in distilled water (0.5 mL) and slowly added at 0°C, and the mixture was stirred for 30 minutes. The mixture was stirred at room temperature for another 30 minutes, and then 2N aqueous hydrochloric acid solution (0.3 mL) was slowly added dropwise to terminate the reaction. The reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC and lyophilized to obtain compound A-34 (0.4 mg, 1%); EI-MS m / z: 825 [M+H] + .

[0912] Example I-31: Preparation of Compound A-35

[0913]

[0914] Step 1: Preparation of compound A-35a

[0915] Core C-9 (572.6 mg, 2.44 mmol) was dissolved in MC (80 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (1.1 g, 2.69 mmol) and benzyltributylammonium chloride (762.5 mg, 2.44 mmol) were added. 5N-sodium hydroxide aqueous solution (1.46 mL, 7.33 mmol) was slowly added to the reaction solution, and the mixture was stirred for 16 hours while the temperature was slowly raised to room temperature. After the reaction was completed, MC (200 mL) and distilled water (200 mL) were added, and the organic layer was extracted three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-35a (1.017 g, 74%) in solid form.

[0916] 1 H-NMR (400MHz, CDCl3) δ7.17-7.10 (m, 2H), 6.80 (dd, J = 7.6, 0.8Hz, 1H), 6.42 (s ,1H),5.57(dd,J=10.4,8Hz,1H),5.48(d,J=3.2Hz,1H),5.15-5.10(m,2H),4.28 -4.23(m,1H),4.20-4.16(m,J=1H),4.10-4.07(m,1H),3.67(s,3H),2.81(t,J= 7.2Hz,2H),2.39(t,J=7.2Hz,2H),2.20(s,3H),2.10-2.05(m,8H),2.03(s,3H).

[0917] Step 2: Preparation of compound A-35b

[0918] Compound A-35a (700 mg, 1.23 mmol) was dissolved in MC (40 mL) at -30 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (450 μL, 4.92 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 4.9 mL, 4.92 mmol) were slowly added in sequence while maintaining the temperature at -10 ° C, and the mixture was stirred for 2 hours. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (100 mL) was added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-35b (190 mg, 25.8%).

[0919] 1H-NMR (400MHz, CDCl3) δ10.33(s,1H),7.69(d,J=8.4Hz,1H),6.88(d,J=8.4Hz,1H),6 .49(s,1H),5.59(dd,J=10.4,8Hz,1H),5.49(d,J=3.6Hz,1H),5.23(d,J=8Hz,1H),5. 16(dd,J=10.4,3.6Hz,1H),4.27-4.14(m,3H),3.68(s,3H),2.89(t,J=7.2Hz,2H),2. 42(t,J=7.2Hz,2H),2.20(s,3H),2.11(m,2H),2.07(s,3H),2.06(s,3H),2.03(s,3H).

[0920] Step 3: Preparation of compound A-35c

[0921] Compound A-35b (190 mg, 0.32 mmol) was dissolved in anhydrous THF (3 mL) at room temperature under a nitrogen atmosphere. Zn powder (CAS No. 7440-66-6, DAEJUNG, 105 mg, 3.2 mmol), 1,2-diiodoethane (CAS No. 624-73-7, Alfa Aesar, 90.4 mg, 0.32 mmol), and propargyl bromide (80% in toluene, 45.5 μL, 0.525 mmol) were then added sequentially. The mixture was sonicated in a Powersonic 410 ultrasonic bath for 30 minutes. After completion of the reaction, the reaction solution was diluted with EA (50 mL), filtered through celite, and extracted with distilled water (50 mL) and 2N aqueous hydrochloric acid (0.7 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-35c (167 mg, 82.6%) as a white solid.

[0922] 1H-NMR(400MHz, CDCl3)δ7.21(m,1H),6.81-6.79(m,1H),6.44(s,1H),5.55(d d,J=10.4,8Hz,1H),5.47(d,J=3.2Hz,1H),5.27(m,1H),5.15-5.09(m,2H),4 .26-4.13(m,2H),4.09(m,1H),3.67(s,3H),2.86-2.78(m,4H),2.57(dd,J=9 .2,4.8Hz,1H),2.40(m,2H),2.19(s,3H),2.08-2.04(m,8H),2.20(s,3H); MS m / z:655[M+Na] + .

[0923] Step 4: Preparation of compound A-35d

[0924] Compound A-35c (42 mg, 0.066 mmol) was dissolved in MC (2 mL) at 0 ° C. under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (27 mg, 0.132 mmol) and pyridine (16 μL, 0.2 mmol) and DIPEA (11.6 μL, 0.066 mmol) were added in sequence, and the mixture was stirred for 1 hour. After the reaction was completed, the reaction solution was diluted with EA (50 mL), and distilled water (50 mL) and 2N-hydrochloric acid aqueous solution (2 mL) were added to extract the organic layer. The extracted organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-35d (51 mg, 96.4%).

[0925] 1 H-NMR (400MHz, CDCl3) δ8.26(d,J=9.2Hz,2H),7.38(d,J=9.2Hz,2H),7.25(m,1H),6.81 (dd,J=8.4,2.4Hz,1H),6.46(s,1H),6.25(m,1H),5.56(dd,J=10.4,8Hz,1H),5.48(d,J= 2.8Hz,1H),5.15-5.12(m,2H),4.27-4.08(m,3H),3.67(s,3H),3.17-3.01(m,2H),2.84 (t,J=7.2Hz,2H),2.41(t,J=7.2Hz,2H),2.20(s,3H),2.11-2.04(m,8H),2.03(s,3H); MS m / z:820[M+Na] + .

[0926] Step 5: Preparation of compound A-35e

[0927] Compound A-35d (32 mg, 0.040 mmol) was dissolved in DMF (3 mL) at 0 ° C. under a nitrogen atmosphere, and then compound PL-1 (13.2 mg, 0.06 mmol), HOBt (5.4 mg, 0.04 mmol), pyridine (300 μL), DIPEA (14 μL, 0.08 mmol) were added in sequence, and the mixture was stirred for 30 minutes and then stirred at room temperature for another 12 hours. After the reaction was completed, EA (50 mL) and 1N-hydrochloric acid aqueous solution (50 mL) were used for extraction. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-35e (27 mg, 77%) as a white solid; MS m / z: 901 [M+Na] + .

[0928] Step 6: Preparation of Compound A-35

[0929] Compound A-35e (27 mg, 0.031 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C, and then lithium hydroxide hydrate (19 mg, 0.461 mmol) dissolved in distilled water (0.2 mL) was added dropwise, and the mixture was stirred for 2.5 hours while the temperature was slowly raised from 0°C to room temperature. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), then separated and purified using preparative HPLC, and lyophilized to obtain compound A-35 (5.8 mg, 27%) as a white solid; EI-MS m / z: 719 [M+Na] + .

[0930] Example I-32: Preparation of Compound A-36

[0931]

[0932] Step 1: Preparation of compound A-36a

[0933] Compound A-25b (277 mg, 0.64 mmol) of Example I-24 was dissolved in MC (21 mL) at 0°C under a nitrogen atmosphere, and then acetyl bromide-α-D-glucuronic acid methyl ester (TCI, CAS No.: 21085-72-3, 405 mg, 1.02 mmol), benzyltributylammonium chloride (199 mg, 0.64 mmol) and 5N-sodium hydroxide aqueous solution (0.53 mL, 2.55 mmol) were slowly added. The mixture was stirred at 0°C for 30 minutes and then at room temperature for 48 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution was added to the reaction solution for neutralization, MC (100 mL) and distilled water (100 mL) were added, and the organic layer was extracted twice. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-36a (144 mg, 30%) as a yellow viscous gum; EI-MS m / z: 751 [M+H] + .

[0934] Step 2: Preparation of compound A-36b

[0935] Compound A-36a (144 mg, 0.19 mmol) was dissolved in THF (10 mL) at 0°C under a nitrogen atmosphere, followed by the addition of sodium borohydride (18 mg, 0.48 mmol) and stirring at 0°C for 3 hours. After completion of the reaction, distilled water (50 mL) was added to terminate the reaction, and the organic layer was extracted twice with EA (50 mL). The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-36b (93.5 mg, 65%) as a colorless viscous gum; EI-MS m / z: 753 [M+H] + .

[0936] Step 3: Preparation of compound A-36c

[0937] Compound A-36b (93.5 mg, 0.12 mmol) was dissolved in MC (5 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (50.1 mg, 0.25 mmol), pyridine (30 μL, 0.37 mmol) and DIPEA (32.4 μL, 0.18 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 30 minutes and stirred at room temperature for another 2.5 hours. After the reaction was completed, the reaction solution was diluted with EA (50 mL), and 2N-hydrochloric acid aqueous solution (50 mL) was added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-36c (80.3 mg, 70%); EI-MS m / z: 918 [M + H] + .

[0938] Step 4: Preparation of compound A-36d

[0939] Compound A-36c (80.3 mg, 0.087 mmol) was dissolved in DMF (1 mL) at 0° C. under a nitrogen atmosphere, and then MMAE (monomethyl auristatin E, CAS No.: 474645-27-7 (62.8 mg, 0.087 mmol), HOBt (17.7 mg, 0.131 mmol), DIPEA (38.1 μL, 0.218 mmol) and pyridine (1 mL) were added sequentially, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted twice with EA (50 mL) and 2N-hydrochloric acid aqueous solution (50 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was separated and purified by preparative HPLC and then lyophilized to obtain compound A-36d (42 mg, 32%) as a white solid; EI-MS m / z: 1497 [M+H] + .

[0940] Step 5: Preparation of Compound A-36

[0941] Compound A-36d (42 mg, 0.028 mmol) was dissolved by mixing and adding methanol (0.7 mL), distilled water (0.35 mL) and THF (0.14 mL) at -20 ° C, and then lithium hydroxide (2.9 mg, 0.07 mmol) dissolved in distilled water (0.35 mL) was added dropwise, and the mixture was stirred for 1 hour while the temperature was slowly raised from -20 ° C to -5 ° C. After the reaction was completed, 2N-hydrochloric acid aqueous solution (0.2 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound A-36 (29.1 mg, 76%) as a white solid; EI-MS m / z: 1357 [M + H] + .

[0942] Example I-33: Preparation of Compound A-37

[0943]

[0944] Step 1: Preparation of compound A-37a

[0945] Compound A-33a (20 mg, 0.034 mmol) was dissolved in DMF (2 mL) at room temperature under a nitrogen atmosphere, and then gefitinib (CAS No.: 18447535-2, 15.2 mg, 0.034 mmol), KI (potassium iodide, 1.7 mg, 0.01 mmol) and DIPEA (14.83 μL, 0.085 mmol) were added to the reaction in sequence, and the mixture was stirred at 60 ° C for 15 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (10 μL) was added for neutralization, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-37a (8.4 mg, 24%); EI-MS m / z: 998 [M + H] + .

[0946] Step 2: Preparation of compound A-37

[0947] Compound A-37a (3.6 mg, 0.0035 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C, and then lithium hydroxide hydrate (1.5 mg, 0.035 mmol) dissolved in distilled water (0.3 mL) was added dropwise, and the mixture was stirred at 0°C for 30 minutes. After the reaction was completed, 2N-hydrochloric acid aqueous solution (10 μL) was added to terminate the reaction. The reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), separated and purified using preparative HPLC, and lyophilized to obtain compound A-37 (1.5 mg, 50%); EI-MS m / z: 830 [M+H] + .

[0948] Example I-34: Preparation of Compound A-38

[0949]

[0950] Step 1: Preparation of compound A-38a

[0951] Compound A-10c (20.0 mg, 0.029 mmol) was dissolved in MC (3.0 mL) at 0°C under a nitrogen atmosphere, and then N,N'-dimethylethylenediamine (TCI, CAS No.: 110-70-3, 31 μL, 0.29 mmol) was added dropwise, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, 2N-hydrochloric acid aqueous solution was slowly added dropwise to adjust the pH of the reaction solution to 7. The reaction solution was separated and purified by preparative HPLC and lyophilized to obtain compound A-38a (8.5 mg, 24%) as a white solid; EI-MS m / z: 625 [M+H] + .

[0952] Step 2: Preparation of compound A-38b

[0953] Compound A-38a (8.5 mg, 0.013 mmol) was dissolved in DMF (2.0 mL) at 0 ° C. under a nitrogen atmosphere, and then compound PL-2 (7.5 mg, 0.013 mmol) and DIPEA (2.37 μL, 0.013 mmol) were added in sequence, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, EA (10 mL) and distilled water (10 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-38b (10.0 mg, 71%) in the form of a clear oil; EI-MS m / z: 1044 [M+H] + .

[0954] Step 3: Preparation of Compound A-38

[0955] Compound A-38b (8.5 mg, 0.013 mmol) was dissolved in methanol (1.0 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (9.27 mg, 0.067 mmol) dissolved in distilled water (500.0 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 2 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC. The product was lyophilized to obtain compound A-38 (1.5 mg, 18%) as a white solid; EI-MS m / z: 875 [M+H] + .

[0956] Example I-35: Preparation of Compound A-39

[0957]

[0958] Step 1: Preparation of compound A-39a

[0959] Core C-3 (55.0 mg, 0.308 mmol) was dissolved in ACN (5.0 mL) at 0°C under a nitrogen atmosphere. 2-(Bromomethyl)-5-nitrofuran (Sigma Aldrich, CAS No. 20782-91-6, 76.3 mg, 0.370 mmol) and potassium carbonate (85.1 mg, 0.616 mmol) were then added sequentially, and the mixture was stirred at room temperature for 16 hours. After the reaction was complete, EA (10 mL) and distilled water (10 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound A-39a (90.0 mg, 96%) as a yellow solid.

[0960] 1 H-NMR (400MHz, CDCl3) δ10.11 (s, 1H), 7.86 (d, J = 8.0Hz, 1H), 7.58 (s, 2H), 7.3 3(d,J=3.6Hz,1H),7.00(d,J=8.0Hz,1H),6.74(d,J=3.6Hz,1H),5.36(s,2H).

[0961] Step 2: Preparation of compound A-39b

[0962] Compound A-39a (90.0 mg, 0.297 mmol) was dissolved in THF (10.0 mL) at 0°C under a nitrogen atmosphere, and sodium borohydride (28.0 mg, 0.742 mmol) was then added, and the mixture was stirred at 0°C for 3 hours. After the reaction was complete, EA (30 mL) and distilled water (30 mL) were added, and the organic layer was extracted. The resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-39b (30.0 mg, 33%) as a yellow solid.

[0963] 1 H-NMR (400MHz, CDCl3) δ7.54(d,J=5.6Hz,1H),7.41(d,J=5.6Hz,1H),7.31-7.28(m,2H),6.81(d ,J=8.0Hz,1H),6.67(d,J=3.6Hz,1H),5.25(s,2H),4.92(d,J=5.6Hz,2H),1.73(t,J=5.6Hz,1H).

[0964] Step 3: Preparation of compound A-39

[0965] Compound A-39b (20.0 mg, 0.065 mmol) and xanthene-9-carboxylic acid (Alfa Aesar, CAS No.: 82-07-5 (16.3 mg, 0.072 mmol) were dissolved in DMF (3.0 mL) at 0°C under a nitrogen atmosphere, and then EDCI hydrochloride (18.8 mg, 0.098 mmol), DMAP (0.8 mg, 0.006 mmol) and DIPEA (34.0 μL, 0.196 mmol) were added in sequence, and the mixture was stirred for 16 hours while slowly raising the temperature from 0°C to room temperature. After completion of the reaction, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), then separated and purified using preparative HPLC, and lyophilized to obtain compound A-39 (2.3 mg, 6.8%) as an ivory solid.

[0966] 1 H-NMR (400MHz, DMSO) δ7.73-7.71(m,2H),7.51(d,J=5.6Hz,1H),7.39-7.32(m,4H),7.26(d,J =8.0Hz,1H),7.18(d,J=8.0Hz,2H),7.15-7.04(m,4H),5.43(s,2H),5.30-5.26(m,3H); EI-MS m / z:536[M+Na] + .

[0967] Example I-36: Preparation of Compound A-40

[0968]

[0969] Compound A-39b (9.1 mg, 0.029 mmol) was dissolved in THF (2.0 mL) at 0°C under a nitrogen atmosphere, and then compound PL-3 and sodium hydride (60% dispersion in mineral oil, 1.44 mg, 0.036 mmol) were added in sequence, and the mixture was stirred at 0°C for 7 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound A-40 (5.1 mg, 2.5%) as a white solid.

[0970] 1 H-NMR (400MHz, CDCl3) δ8.49(s,1H),8.33(s,1H),7.51(d,J=8.0Hz,1H),7.41(d,J=5.6Hz,1 H),7.28(d,J=5.6Hz,1H),7.22-7.18(m,2H),7.13-7.09(m,1H),6.68(d,J=8.0Hz,1H),6.55( d,J=3.6Hz,1H),5.86(s,1H),5.31-5.30(m,1H),5.24(s,2H),5.12(s,2H),4.40-4.37(m,1H EI-MS m / z:681[M+H] + .

[0971] Example I-37: Preparation of Compound A-41

[0972]

[0973] Step 1: Preparation of compound A-41a

[0974] Compound A-10c (20.0 mg, 0.029 mmol) was dissolved in DMF (3.0 mL) at 0°C under a nitrogen atmosphere, and then doxorubicin (Sigma Aldrich, CAS No.: 23214-92-8, 19.3 mg, 0.035 mmol) and DIPEA (7.6 μL, 0.043 mmol) were added in sequence, and the mixture was stirred at 0°C for 4 hours. After the reaction was completed, EA (20 mL) and distilled water (20 mL) were added, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-41a (12.0 mg, 38%) as a red solid; EI-MS m / z: 1102 [M+Na] + .

[0975] Step 2: Preparation of compound A-41

[0976] Compound A-41a (12.0 mg, 0.011 mmol) was dissolved in methanol (1.0 mL) and THF (0.5 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (10.7 mg, 0.077 mmol) dissolved in distilled water (200.0 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 2 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-41 (0.3 mg, 2.9%) as a red solid; EI-MS m / z: 934 [M+Na] + .

[0977] Example I-38: Preparation of Compound A-42

[0978]

[0979] Step 1: Preparation of compound A-42a

[0980] Compound A-10c (18.2 mg, 0.027 mmol) was dissolved in DMF (2.0 mL) under a nitrogen atmosphere, and then lapatinib (TCI, CAS No.: 231277-92-2, 15.6 mg, 0.027 mmol) and DIPEA (12.0 μL, 0.067 mmol) were added dropwise in sequence, and the mixture was stirred at room temperature for 48 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC. The product was lyophilized to give compound A-42a (7.0 mg, 23%) as a white solid; EI-MS m / z: 1118 [M+H] + .

[0981] Step 2: Preparation of Compound A-42

[0982] Compound A-42a (7.0 mg, 0.006 mmol) was dissolved in methanol (1.0 mL) and THF (0.5 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (6.0 mg, 0.044 mmol) dissolved in distilled water (200.0 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 30 minutes. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-42 (3.7 mg, 62%) as a white solid; EI-MS m / z: 949 [M+H] + .

[0983] Example I-39: Preparation of Compound A-43

[0984]

[0985] Step 1: Preparation of compound A-43a

[0986] Compound A-10b (69.0 mg, 0.135 mmol) was dissolved in MC (2.0 mL) at 0 ° C under a nitrogen atmosphere, and then thionyl chloride (21.6 μL, 0.296 mmol) was slowly added dropwise, and the mixture was stirred at 0 ° C for 3 hours. After the reaction was completed, MC (20 mL) and distilled water (20 mL) were added to extract the organic layer. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-43a (62.0 mg, 87%) as a white solid.

[0987] 1H-NMR (400MHz, CDCl3) δ7.46-7.42(m,2H),7.30(d,J=8.0Hz,1H),6.93(d,J=8.0Hz,1H),5.63(dd,J=10.4Hz,J=8.0Hz,1H),5.50(d,J=3.2Hz 1H),5.17-5.13(m,2H),4.83(s,2H),4.28-4.24(m,1H),4.21-4.18(m,1H ),4.16-4.11(m,1H),2.20(s,3H),2.08(s,3H),2.04(s,3H),2.03(s,3H).

[0988] Step 2: Preparation of compound A-43b

[0989] Compound A-43a (21.0 mg, 0.040 mmol) was dissolved in DMF (1.0 mL) at 0°C under a nitrogen atmosphere, and then erlotinib (Sigma-Aldrich, CAS No.: 183319-69-9, 20.6 mg, 0.048 mmol) and potassium iodide (0.6 mg, 0.004 mmol) were added dropwise in sequence, and the mixture was stirred at 80°C for 16 hours. After completion of the reaction, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-43b (2.2 mg, 62%) as a white solid.

[0990] 1 H-NMR (400MHz, DMSO) δ8.25(s,1H),7.80(d,J=5.6Hz,1H),7.64(s,1H),7.40(d,J=4.4Hz,1H),7.2 7-7.23(m,2H),7.12(s,1H),7.07-7.04(m,3H),6.77(s,1H),5.57-5.53(m,3H),5.39-5.31(m,3H) ,4.47(t,J=6.4Hz,1H),4.13-4.12(m,4H),4.07(s,1H),3.94(t,J=4.4Hz,2H),3.66(t,J=4.4Hz,2 EI-MS m / z:887[M+H] + .

[0991] Step 3: Preparation of Compound A-43

[0992] Compound A-43b (5.0 mg, 0.0056 mmol) was dissolved in methanol (1.0 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (5.45 mg, 0.039 mmol) dissolved in distilled water (200.0 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 2 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound A-43 (2.2 mg, 54%) as a white solid.

[0993] 1 H-NMR (400MHz, DMSO) δ8.29 (s, 1H), 8.26 (s, 1H), 7.71 (d, J = 5.6Hz, 1H), 7.66-7.63 (m ,2H),7.34(d,J=8.0Hz,1H),5.67(t,J=8.0Hz,1H),7.13-7.09(m,2H),7.08-7.03(m, 2H),6.81-6.77(m,1H),5.55(s,2H),4.89(d,J=8.0Hz,1H),4.11(t,J=4.4Hz,2H),4. 06(s,1H),3.93(t,J=4.4Hz,2H),3.73-3.44(m,9H),3.29(s,3H),3.18(s,3H); EI-MS m / z:719[M+H] + .

[0994] Example I-40: Preparation of Compound A-44

[0995]

[0996] Step 1: Preparation of compound A-44a

[0997] Compound A-43a (36.2 mg, 0.068 mmol) was dissolved in DMF (1.0 mL) at 0°C under a nitrogen atmosphere, and imatinib (TCI, CAS No.: 152459-95-5, 40.5 mg, 0.082 mmol) and potassium iodide (1.1 mg, 0.006 mmol) were added dropwise in sequence, and the mixture was stirred at 80°C for 16 hours. After completion of the reaction, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-44a (18.0 mg, 26%) as a white solid.

[0998] 1 H-NMR (400MHz, DMSO) δ10.24(s,1H),9.27(d,J=3.2Hz,1H),8.99(s,1H),8.68(dd,J=8.0Hz,J=3.2Hz 1H),8.54(s,1H),8.51(d,J=5.6Hz,1H),8.47(dt,J=8.0Hz,J=3.2Hz,1H),8.08(s,1H),7.96(d,J=8.0Hz,2H),7.89(d,J=5.6H z,1H),7.62(d,J=8.4Hz,1H),7.53-7.42(m,5H),7.29(d,J=5.6Hz,1H),7.21(d,J=8.4Hz,1H),7.15(d,J=8.4Hz,1H),5.65(d, J=10.8Hz,1H),5.40-5.35(m,3H),4.86(s,2H),4.55(t,J=6.4Hz,1H),4.16-4.13(m,2H),3.71(s,2H),3.65-3.62(m,2H),3.5 1-3.48(m,2H),3.06(s,3H),2.91-2.88(m,2H),2.74-2.68(m,2H),2.22(s,3H),2.16(s,3H),2.03(s,3H),1.98(s,3H); EI-MS m / z:988[M+H] + ,4941 / 2[M+H] + .

[0999] Step 2: Preparation of Compound A-44

[1000] Compound A-44a (18.0 mg, 0.018 mmol) was dissolved in methanol (1.0 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (17.6 mg, 0.127 mmol) dissolved in distilled water (200.0 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 30 minutes. After completion of the reaction, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to give compound A-44 (7.0 mg, 46%) as a white solid.

[1001] 1H-NMR (400MHz, DMSO) δ10.23(s,1H),9.27(d,J=1.6Hz,1H),8.98(s,1H),8.68(dd,J=5.6Hz,J=1.6Hz,1H),8.51(d,J=5.6Hz,1H ),8.48-8.46(m,2H),8.08(s,1H),7.96(d,J=5.6Hz,2H),7.80(d,J=5.6Hz,1H),7.66(d,J=5.6Hz,1H),7.56-7.42(m,5H),7.23- 7.18(m,2H),4.99(d,J=7.6Hz,1H),4.85(s,2H),3.76-3.46(m,15H),3.05(s,3H),2.91-2.88(m,2H),2.74-2.69(m,2H),2.22(s ,3H),3.17(s,2H),3.65-3.62(m,2H),3.51-3.48(m,2H),3.06(s,3H),2.91-2.88(m,2H),2.74-2.68(m,2H),2.22(s,3H); EI-MS m / z:987[M+H] + ,494 1 / 2[M+H] + .

[1002] Example I-41: Preparation of Compounds A-45 and A-50

[1003]

[1004] Compound A-25 (13.5 mg, 0.01 mmol) was dissolved in THF (1 mL) at room temperature under a nitrogen atmosphere, followed by the addition of triphenylphosphine (3.95 mg, 0.015 mmol) and distilled water (0.3 mL), and the mixture was stirred for 12 hours. A 2N aqueous sodium hydroxide solution (10 μL) was added to the reaction solution at room temperature, and the mixture was stirred for 30 minutes. A 2N aqueous hydrochloric acid solution was then slowly added dropwise to adjust the pH of the reaction solution to 3. The reaction solution was separated and purified by preparative HPLC and then lyophilized to obtain compounds A-45 and A-50 (A-45: 1 mg, 7%; A-50: 7.8 mg, 59%), respectively; MS m / z: A-45 = 1317 [M+H] + ,A-50=1318[M+H] + .

[1005] Example I-42: Preparation of Compound A-46

[1006]

[1007] Step 1: Preparation of compound A-46a

[1008] Compound A-26g (50.0 mg, 0.070 mmol) was dissolved in DMF (2.0 mL) at 0 ° C. under a nitrogen atmosphere, and then MMAE (50.0 mg, 0.070 mmol), HOBt (14.0 mg, 0.105 mmol), DIPEA (30.0 μL, 0.175 mmol) and pyridine (1.0 mL) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, it was extracted with distilled water (50 mL), 2N-hydrochloric acid aqueous solution (50 mL) and EA (100 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-46a (75.0 mg, 83%) as a white solid; EI-MS m / z: 1294 [M+H] + .

[1009] Step 2: Preparation of compound A-46

[1010] Compound A-46a (62.0 mg, 0.048 mmol) was dissolved in methanol (1.0 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (46.3 mg, 0.335 mmol) dissolved in distilled water (1.0 mL) was slowly added dropwise, and the mixture was stirred at 0°C for 1 hour. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), separated and purified using preparative HPLC, and lyophilized to obtain compound A-46 (29.0 mg, 59%) as a white solid; EI-MS m / z: 1126 [M+H] + .

[1011] Example I-43: Preparation of Compound A-47

[1012]

[1013] Compound A-46 (12.4 mg, 0.011 mmol) was dissolved in THF (2 mL) at room temperature under a nitrogen atmosphere, and triphenylphosphine (4.33 mg, 0.0165 mmol) and distilled water (0.1 mL) were added in sequence, and the mixture was stirred for 48 hours. After the reaction was completed, the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), and then separated and purified by preparative HPLC, and then lyophilized to obtain compound A-47 (3 mg, 24.8%); EI-MS m / z: 1100 [M+H] + .

[1014] Example I-44: Preparation of Compound A-48

[1015]

[1016] Step 1: Preparation of compound A-48a

[1017] Compound A-26g (43.0 mg, 0.060 mmol) was dissolved in MC (6.0 mL) at 0°C under a nitrogen atmosphere, and then N,N'-dimethylethylenediamine (TCI, CAS No.: 110-70-3, 64 μL, 0.6 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC. The product was lyophilized to obtain compound A-48a (28.5 mg, 71%) as a white solid; EI-MS m / z: 664 [M+H] + .

[1018] Step 2: Preparation of compound A-48b

[1019] Compound A-48a (28.5 mg, 0.043 mmol) was dissolved in DMF (1.0 mL) at 0°C under a nitrogen atmosphere, and then compound PL-2 (24 mg, 0.043 mmol) and DIPEA (7.5 μL, 0.043 mmol) were added in sequence, and the mixture was stirred at room temperature for 4 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction. The reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC. The product was lyophilized to give compound A-48b (26.5 mg, 57%) as a white solid; EI-MS m / z: 1083 [M+H] + .

[1020] Step 3: Preparation of Compound A-48

[1021] Compound A-48b (26.5 mg, 0.024 mmol) was dissolved in methanol (1.0 mL) and THF (0.5 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (23.6 mg, 0.171 mmol) dissolved in distilled water (200.0 μL) was slowly added dropwise, and the mixture was stirred at 0°C for 2 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1.0 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1.0 mL) and distilled water (1.0 mL), and then separated and purified using preparative HPLC, and lyophilized to obtain compound A-48 (16.5 mg, 73%) as a white solid; EI-MS m / z: 914 [M+H] + .

[1022] Example I-45: Preparation of Compound A-49

[1023]

[1024] Step 1: Preparation of compound A-49a

[1025] LAH (2.25 g, 59.29 mmol) was added to THF (300 mL) at room temperature under a nitrogen atmosphere, and the mixture was then cooled to 0°C. Compound C-15e (8.53 g, 23.74 mmol) was dissolved in THF (75 mL) and slowly added, and the mixture was stirred at 0°C for 5 minutes. After the reaction was completed, 2N-sodium hydroxide aqueous solution (36 mL) was added to terminate the reaction, and the reaction solution was diluted with THF (150 mL) and filtered through diatomaceous earth. The resulting solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-49a (7.2 g, 96%) as an ivory solid.

[1026] 1 H-NMR (400MHz, CDCl3) δ7.66 (s, 1H), 7.48-7.33 (m, 6H), 7.19 (t, J = 8Hz, 1H), 6 .83(d,J=8Hz,1H),5.19(s,2H),4.93(d,J=6Hz,2H),1.92(t,J=6Hz,1H); EI-MS m / z:318[M+H] + .

[1027] Step 2: Preparation of compound A-49b

[1028] Compound A-49a (4.66 g, 14.69 mmol) was added to MC (93 mL) under a nitrogen atmosphere at room temperature, followed by the addition of TEMPO (Alfa aesar, CAS No.: 2564-83-2, 230 mg, 1.47 mmol) and TBAI (543 mg, 1.47 mmol). Sodium bicarbonate (4.2 g, 0.5 mol) and potassium carbonate (691 mg, 0.05 mol) were then dissolved in 100 mL of water, followed by the addition of NCS (N-chlorosuccinimide, Merck, CAS No.: 128-09-6, 2.16 g, 16.18 mmol), and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, MC (100 mL) and distilled water (100 mL) were added, and the organic layer was extracted twice. The organic layer was washed by adding a saturated sodium chloride aqueous solution (100 mL), and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-49b (4.35 g, 94%) as a yellow solid.

[1029] 1 H-NMR (400MHz, CDCl3) δ9.95 (s, 1H), 8.53 (s, 1H), 7.53-7.35 (m, 7H), 6.88 (d, J = 8Hz, 1H), 5.23 (s, 2H); EI-MS m / z: 316 [M+H] + .

[1030] Step 3: Preparation of compound A-49c

[1031] Compound A-49b (4.35 g, 13.80 mmol) was dissolved in MC (70 mL) at room temperature under a nitrogen atmosphere. (Carboethoxymethylene)triphenylphosphorane (Merck, CAS No. 1099-45-2, 9.16 g, 27.59 mmol) was then added, and the mixture was stirred at room temperature for 1.5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to afford compound A-49c (5.12 g, 96%) as an ivory solid.

[1032] 1 H-NMR (400MHz, CDCl3) δ7.94 (s, 1H), 7.86 (d, J = 15.2Hz, 1H), 7.48-7.34 (m, 6H), 6.82 (d, J = 8Hz ,1H),6.10(d,J=15.2Hz,1H),5.20(s,2H),4.26(q,J=7.2Hz,2H),1.33(t,J=7.2Hz,3H); EI-MS m / z:386[M+H] + .

[1033] Step 4: Preparation of compound A-49d

[1034] Compound A-49c (5.12 g, 13.29 mmol) was dissolved in EA (147 mL) and methanol (441 mL) at room temperature under a nitrogen atmosphere, and 5% palladium on carbon (5% Pd / C, 5.12 g) was added, and the mixture was stirred for 16 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was diluted with MC (800 mL) and filtered through diatomaceous earth. The filtered solution was concentrated under reduced pressure. The resulting residue obtained by column chromatography gave compound A-49d (4.53 g, 88%) in the form of a beige oil.

[1035] 1H-NMR (400MHz, CDCl3) δ7.50-7.3(m,7H),7.14(t,J=8Hz,1H),6.81(d,J=8Hz,1H),5.12(s,2H), 4.16(q,J=7.2Hz,2H),3.25(t,J=7.6Hz,2H),2.73(t,J=7.6Hz,2H),1.26(t,J=7.2Hz,3H); EI-MS m / z:388[M+H] + .

[1036] Step 5: Preparation of compound A-49e

[1037] Compound A-49d (3.88 g, 10.01 mmol) was dissolved in MC (287 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (Merck, CAS No.: 4885-02-3, 2.72 mL, 30.01 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 30 mL, 30 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 2 hours. After the reaction was completed, distilled water (250 mL) was slowly added dropwise to terminate the reaction, and MC (250 mL) was added to extract the organic layer twice. The organic layer was washed by adding a saturated sodium chloride aqueous solution (250 mL), and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-49e (2.59 g, 62%) in the form of a yellow oil.

[1038] 1 H-NMR (400MHz, CDCl3) δ10.05(s,1H),7.72(d,J=8.4Hz,1H),7.58(s,1H),7.49-7.36(m,5H),6.99(d,J=8.4Hz,1 EI-MS m / z:416[M+H] + .

[1039] Step 6: Preparation of Compound A-49f

[1040] Compound A-49e (2.85 g, 6.86 mmol) was dissolved in ethanol (17 mL) and THF (17 mL) at 0 ° C under a nitrogen atmosphere, and then lithium hydroxide monohydrate (864 mg, 20.59 mmol) dissolved in distilled water (8.5 mL) was slowly added dropwise, and the mixture was stirred at room temperature for 1.5 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (15 mL) was added to terminate the reaction, and the organic layer was extracted with EA (85 mL) and distilled water (85 mL). The organic layer was washed by adding saturated sodium chloride aqueous solution (250 mL), and the resulting organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue obtained by column chromatography gave compound A-49f (2.56 g, 96%) in the form of an orange solid.

[1041] 1 H-NMR (400MHz, CDCl3) δ10.05 (s, 1H), 7.73 (d, J = 8Hz, 1H), 7.60 (s, 1H), 7.52-7.36 (m, 5H ),6.99(d,J=8.4Hz,1H),5.31(s,2H),3.33(t,J=7.6Hz,2H),2.83(t,J=7.6Hz,2H); EI-MS m / z:388[M+H] + .

[1042] Step 7: Preparation of Compound A-49g

[1043] Compound A-49f (2.56 g, 6.61 mmol) was dissolved in DMF (22 mL) at 0 ° C under a nitrogen atmosphere, and then EDC (1.9 g, 9.91 mmol), HOBt (894 mg, 6.61 mmol), DMAP (81 mg, 0.66 mmol) and DIPEA (3.45 mL, 19.81 mmol) were added in sequence, and the mixture was stirred for 1.5 hours. In addition, linker P-1 (1.68 g, 6.60 mmol) dissolved in DMF (11 mL) was slowly added dropwise, and the reaction mixture was stirred at room temperature for 16 hours, and then extracted with EA (100 mL) and distilled water (100 mL). The obtained organic layer was washed by adding saturated sodium chloride aqueous solution (50 mL), then dried over anhydrous magnesium sulfate, and the organic layer was filtered and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-49g (2.24 g, 58%) in the form of a brown oil.

[1044] 1H-NMR (400MHz, CDCl3) δ10.04(s,1H),7.71(d,J=8.4Hz,1H),7.58(s,1H),7.49-7.36(m,5H),6.98(d,J=8Hz,1H),6 .14(brs,1H),5.30(s,2H),3.71-3.49(m,14H),3.47-3.44(m,2H),3.36-3.33(m,4H),2.61(t,J=7.6Hz,2H); EI-MS m / z:588[M+H] + .

[1045] Step 8: Preparation of Compound A-49h

[1046] Compound A-49g (2.23 g, 3.80 mmol) was dissolved in MC (76 mL) at -78 ° C under a nitrogen atmosphere, and then pentamethylbenzene (Alfa aesar, CAS No.: 700-12-9, 1.69 g, 11.40 mmol) was added, and a boron trichloride solution (1M-BCl3 in MC, 22.8 mL, 22.8 mmol) was slowly added, and the mixture was stirred for 1 hour while slowly raising the temperature to -55 ° C. After the reaction was completed, distilled water (150 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C for neutralization. MC (150 mL) and distilled water (150 mL) were added to the mixture, and the organic layer was extracted twice. The resulting organic layer was washed by adding a saturated sodium chloride aqueous solution (150 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-49h (1.33 g, 71%) as a beige solid.

[1047] 1 H-NMR (400MHz, CDCl3) δ10.01(s,1H),8.42(brs,1H),7.66-7.64(m,2H),6.94(d,J=8Hz,1H),6.32(brs,1H EI-MS m / z:498[M+H] + .

[1048] Step 9: Preparation of compound A-49i

[1049] Compound A-49h (500 mg, 1.01 mmol) was dissolved in MC (33 mL) at 0°C under a nitrogen atmosphere, and then acetyl bromide-α-D-glucuronic acid methyl ester (TCI, 838 mg, 2.11 mmol), benzyltributylammonium chloride (314 mg, 1.01 mmol) and 5N-sodium hydroxide aqueous solution (1 mL, 5.00 mmol) were slowly added. The mixture was stirred at 0°C for 30 minutes and then stirred at room temperature for another 16 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution was added to the reaction solution for neutralization, and MC (165 mL) and distilled water (165 mL) were added for extraction. The resulting organic layer was washed by adding saturated sodium chloride aqueous solution (165 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-49i (200 mg, 25%) in the form of an ivory-colored viscous gum.

[1050] 1 H-NMR (400MHz, CDCl3) δ10.10(s,1H),7.73(d,J=8Hz,1H),7.39(s,1H),7.09(d,J=8.4Hz,1H),6.16(brs,1H),5.44-5.37(m, 4H), 4.29 (m, 1H), 3.72 (s, 3H), 3.67-3.46 (m, 14H), 3.38-3.33 (m, 4H), 2.61 (t, J = 7.6Hz, 2H), 2.09 (s, 3H), 2.07 (s, 6H); EI-MS m / z:814[M+H] + .

[1051] Step 10: Preparation of compound A-49j

[1052] Compound A-49i (198 mg, 0.24 mmol) was dissolved in THF (13.3 mL) at 0 ° C under a nitrogen atmosphere, and then sodium borohydride (NaBH4, 27.6 mg, 0.73 mmol) was added, and the mixture was stirred at 0 ° C for 5 hours. After the reaction was completed, distilled water (13 mL) was added to terminate the reaction, and the organic layer was extracted twice with EA (53 mL) and distilled water (27 mL). The resulting organic layer was washed with saturated sodium chloride solution (53 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-49j (146 mg, 74%) in the form of a white viscous gel.

[1053] 1H-NMR (400MHz, CDCl3) δ7.34(s,1H),7.14(d,J=8Hz,1H),6.93(d,J=8Hz,1H),6.22(brs,1H),5.39-5.35(m,3H),5.19(m,1H),4.8 1(d,J=5.6Hz,2H),4.20(m,1H),3.74(s,3H),3.65-3.43(m,14H),3.38-3.28(m,4H),2.61(m,2H),2.07(s,3H),2.05(s,6H); EI-MS m / z:816[M+H] + .

[1054] Step 11: Preparation of Compound A-49k

[1055] Compound A-49j (171 mg, 0.21 mmol) was dissolved in MC (5.1 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (85 mg, 0.42 mmol), pyridine (50.7 μL, 0.63 mmol) and DIPEA (54.8 μL, 0.32 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 30 minutes and stirred at room temperature for another 2.5 hours. After the reaction was completed, the reaction solution was diluted with EA (25 mL), and the organic layer was extracted by adding 2N-hydrochloric acid aqueous solution (25 mL). The obtained organic layer was washed by adding saturated sodium chloride aqueous solution (25 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to give compound A-49k (158 mg, 77%) in the form of a transparent viscous foam.

[1056] 1 H-NMR (400MHz, CDCl3) δ8.27(d,J=8.8Hz,1H),7.41-7.37(m,3H),6.95(d,J=8.4Hz,1H),6.20(brs,1H),5.43-5.37(m,5H),5.24-5.23(m,1H EI-MS m / z:981[M+H] + .

[1057] Step 12: Preparation of Compound A-49L

[1058] Compound A-49k (41.1 mg, 0.042 mmol) was dissolved in DMF (1 mL) at 0 ° C. under a nitrogen atmosphere, and then MMAE (30 mg, 0.042 mmol), HOBt (8.5 mg, 0.063 mmol), DIPEA (18.2 μL, 0.104 mmol) and pyridine (1 mL) were added sequentially, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted twice with EA (20 mL) and 2N-hydrochloric acid aqueous solution (20 mL). The obtained organic layer was washed by adding saturated sodium chloride aqueous solution (20 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-49L (47.7 mg, 73%) as a white solid; EI-MS m / z: 1560 [M+H] + ,7801 / 2[M+H] + .

[1059] Step 13: Preparation of Compound A-49

[1060] Compound A-49L (47.7 mg, 0.031 mmol) was dissolved by adding methanol (0.76 mL), distilled water (0.1 mL) and THF (0.48 mL) at -20 ° C., and then lithium hydroxide (5.1 mg, 0.12 mmol) dissolved in distilled water (0.44 mL) was added dropwise, and the mixture was stirred at -20 ° C for 1 hour. After the reaction was completed, 2N-hydrochloric acid aqueous solution (1 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL). It was separated and purified using preparative HPLC and lyophilized to obtain compound A-49 (28.2 mg, 65%) as a white solid; EI-MS m / z: 1420 [M+H] + ,7101 / 2[M+H] + .

[1061] Example I-46: Preparation of Compound A-51

[1062]

[1063] Step 1: Preparation of compound A-51a

[1064] Compound A-49k (37 mg, 0.038 mmol) was dissolved in DMF (1 mL) at 0 ° C. under a nitrogen atmosphere, and then MMAF-OMe (28 mg, 0.038 mmol), HOBt (7.6 mg, 0.056 mmol), DIPEA (16.4 μL, 0.094 mmol) and pyridine (1 mL) were added sequentially, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the organic layer was extracted twice with EA (20 mL) and 2N-hydrochloric acid aqueous solution (20 mL). The obtained organic layer was washed by adding saturated sodium chloride aqueous solution (20 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-51a (45 mg, 75%) in the form of a transparent viscous foam; EI-MS m / z: 1588 [M+H] + ,794 1 / 2[M+H] + .

[1065] Step 2: Preparation of compound A-51

[1066] Compound A-51a (45 mg, 0.028 mmol) was dissolved by mixing and adding methanol (0.71 mL), distilled water (0.11 mL) and THF (0.14 mL) at -20 ° C, and then lithium hydroxide (8.9 mg, 0.212 mmol) dissolved in distilled water (0.6 mL) was added dropwise, and the mixture was stirred for 3 hours while the temperature was slowly raised from -20 ° C to -5 ° C. After the reaction was completed, 2N-hydrochloric acid aqueous solution (0.5 mL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), separated and purified using preparative HPLC, and lyophilized to give compound A-51 (22.2 mg, 55%) as a white solid; EI-MS m / z: 1434 [M + H] + ,717 1 / 2[M+H] + .

[1067] Example I-47: Preparation of Compound A-52

[1068]

[1069] Step 1: Preparation of compound A-52a

[1070] Compound A-49b (8 g, 25.38 mmol) was dissolved in MC (242 mL, 0.105 M) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1 M-BCl3 in MC, 76.14 mL, 76.14 mmol) was slowly added, and the mixture was stirred for 2 hours. After the reaction was completed, distilled water (200 mL) was added to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize it. 2N-hydrochloric acid aqueous solution was slowly added dropwise to the mixture, and the pH value of the reaction solution was adjusted to 7. MC (200 mL) was added to the mixture and the organic layer was extracted twice. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue obtained by column chromatography gave compound A-52a (4.9 g, 85.8%) in the form of a yellow-green solid.

[1071] 1 H-NMR (400MHz, DMSO) δ9.97 (s, 1H), 8.65 (s, 1H), 7.53 (d, J = 8Hz, 1H), 7.32 (t, J = 8Hz, 1H), 6.82 (d, J = 8Hz, 1H); EI-MS m / z: 226 [M+H] + .

[1072] Step 2: Preparation of compound A-52b

[1073] Silver carbonate (9.07 g, 32.9 mmol) and HMTTA (1,1,4,7,10,10-hexamethyltriethylenetetramine, 1.13 g, 4.88 mmol) were dissolved in ACN (80 mL) at room temperature under a nitrogen atmosphere, and the mixture was stirred for 1 hour. The reaction solution was cooled to 0 ° C, and then a solution of compound A-52a (2 g, 8.88 mmol) dissolved in ACN (30 mL) and a solution of methyl acetyl bromide-α-D-glucuronide (TCI, 8.82 g, 22.2 mmol) dissolved in ACN (20 mL) were slowly added in sequence, and the mixture was stirred for 16 hours while slowly raising the temperature from 0 ° C to room temperature. After the reaction was completed, the filtrate obtained by the diatomaceous earth filter was concentrated under reduced pressure. EA (100 mL) and distilled water (100 mL) were added to the concentrate thus obtained, and the organic layer was extracted twice. The obtained organic layer was washed by adding a sodium chloride aqueous solution. The washed organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-52b (0.82 g, 17%) as a yellow solid. [1074...

Claims

1. A compound comprising a self-destructing group represented by the following formula 1, or a pharmaceutically acceptable salt thereof: [Formula 1] A-(L 1 ) k -U j In formula 1, L 1 is a divalent or multivalent linking group; k is 0 or 1, j is 1 to 10; A is absent, H or a bound functional group; and U is a part represented by the following formula A: [Formula A] In formula A, R 1 and R 2 Each independently is H or C 1-8 Saturated or unsaturated hydrocarbon group; PL is an active agent which is bonded to L via a heteroatom selected from N, O and S. 2 or with R 1 and R 2 The bonded carbon atoms are connected; L 2 is a self-eliminating joint, which is selected so that L 2 With R 1 and R 2 The cleavage of the bond between the bound carbon atoms promotes L 2 Cleavage of the bond with PL; W is an optional substituent on the benzene ring; Z 1 and Z 3 One selected from N, NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 are each independently CH or N, and -(L 1 )kA and -(V)h, if present, each independently replace H in NH or CH; Indicates that A-(L 1 )k-formed bonds; R 3 H or C 1-8 Hydrocarbon; V is an electron-withdrawing group or an electron-donating group; T is a trigger group capable of initiating the release of PL and L2 if present, via a 1,6-elimination reaction upon cleavage; L 3 is an optional self-destructive spacer group, which, if present, is cleaved sequentially when T is cleaved; X and Y are each independently selected from -O-, -NH-, and S; and h, i, l, x and y are each independently 0 or 1, and p is an integer from 0 to 2.

2. The compound comprising a self-destructing group according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The ring in formula A Selected from the following groups:

3. The compound comprising a self-destructing group according to claim 1, wherein V is selected from the group consisting of halogen, CN, NO2, formyl, C 1-8 Alkylcarbonyl, carboxyl, C 1-8 Alkoxycarbonyl, carboxy-C1-8alkyl, carbamoyl, mono-C 1-8 Alkylcarbamoyl, di-C 1-8 Alkylcarbamoyl, C 1-8 Alkyl, C 1-8 Alkenyl, OH, C 1-8 Alkoxy, SH, C 1-8 Alkylthiol, NH2, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino and C 6-18 Aryl.

4. The compound comprising a self-destructing group according to claim 1, wherein W is selected from the group consisting of: H, C 1-12 Saturated or unsaturated hydrocarbon, halogen, halogenated-C 1-8 Alkyl, CN, NO2, OH, C 1-8 Alkoxy, hydroxyl C 1-8 Alkyl, C 1-8 Alkoxy-C 1-8 Alkyl, SH, C 1-8 Alkylthio, mercapto-C 1-8 Alkyl, amino, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino, amino-C 1-8 Alkyl, C 1-8 Monoalkylamino-C 1-8 Alkyl, C 1-8 Dialkylamino-C 1-8 Alkyl, carboxyl, C 1-8 Alkoxycarbonyl, C 1-8 Alkoxycarbonyloxy, carboxyl-C 1-8 Alkyl, C 1-8 Alkoxycarbonyl-C 1-8 Alkyl, carbamoyl, mono-C 1-8 Alkylcarbamoyl, di-C 1-8 Alkylcarbamoyl, carbamoyl-C 1-8 Alkyl, mono-C 1-8 Alkylcarbamoyl-C 1-8 Alkyl and di-C 1-8 Alkylcarbamoyl-C 1-8 alkyl.

5. The compound comprising a self-destructing group according to claim 1, wherein -(Y) y- T is selected from the group consisting of: -O-SO3 - ;-NO2;-OC(O)(CH2) r COR t1 ; -O(CH2)-Ar 1 -NO2; -SC(O)(CH2) s COR t2 ; -S(CH2)-Ar 2 -NO2 and -BR t3 R t4 The part represented; in The -OH group may be protected by a protecting group or substituted by a substituent; R t1 and R t2 is a C1-C8 alkyl group, Ar 1 and Ar 2 C5-C 20 an arylene or heteroarylene group; R t3 and R t4 Each is independently hydrogen, C1-C8 alkoxy or hydroxy; R t5 OH, single-C 1-8 Alkylamino, di-C 1-8 Alkylamino or -NH(CH2CH2O) f R t6 , where R t6 H or C 1-4 Alkyl; and f is an integer of 1-10, and r and s are integers of 1-5 each.

6. The compound comprising a self-destructing group according to claim 1, wherein -(X) x -L 3 -for and R 8 and R 9 Each independently selected from H, halogen, C 1-8 The present invention is a group consisting of alkyl, CN and NO2, and o is an integer from 0 to 2.

7. The compound comprising a self-destructing group according to claim 1, wherein PL is an active agent selected from a drug, a toxin, a fluorophore, an affinity ligand, a diagnostic substance or a detection probe.

8. The compound comprising a self-destructing group according to claim 7, wherein The drug is selected from the group consisting of a cytokine, an immunomodulatory compound, an anticancer agent, an antiviral agent, an antibacterial agent, an antifungal agent, an anthelmintic, or a combination thereof.

9. The compound comprising a self-destructing group according to claim 1, wherein The PL is selected from the group consisting of:

10. The compound comprising a self-destructing group according to claim 1, wherein L 2 Selected from -OC(=O)-, -S(=O)2-, The group composed of R 10 To R 12 Each independently is H, C 1-8 Alkyl, amino-C 1-8 Alkyl, mono-(C 1-8 alkyl)amino or di-(C 1-8 Alkyl)amino substituted C 1-8 Alkyl, or -(CH2CH2O) g R 13 ,in R 13 H or C 1-4 An alkyl group is selected from the group consisting of:

11. The compound comprising a self-destructing group according to claim 1, wherein The binding functional group is a functional group that can bind to a ligand or protein having receptor binding properties, or a linker precursor through a click chemistry reaction.

12. The compound comprising a self-destructing group according to claim 1, wherein The binding functional group comprises a functional group selected from the group consisting of halogen, OH, C 1-8 Alkoxy, hydroxyamino, COH, C1-8 alkylcarbonyl, carboxyl, C 1-8 Alkoxycarbonyl, tosyl, tosylate, amino, mono-C 1-8 Alkylamino, di-C 1-8 Alkylamino, NHNH2, N3, haloacetamide, maleimide, succinimide, SH, SO3H, C 1-8 Alkylsulfonyl, C 1-8 Alkoxysulfonyl, 2-pyridyl disulfide, PO3H2, OPO3H2, -N≡C, -NCS, C 4-10 Dienyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 4-10 Cycloalkynyl and C 2-8 Alkynylcarbonyl, and R f H or C 1-8 alkyl.

13. The compound comprising a self-destructing group according to claim 1, wherein AL 1 The precursor of A and L 1 The precursor of A comprises at least one functional group selected from the group consisting of: hydroxyl, amino, azido, alkynyl, conjugated diene, alkenyl, cyclooctynyl, maleimide, SO2N3, alkoxysulfinyl, oxiranyl, aziridine, oxo, hydrazine, hydroxyamino, thiol and 1,3-dicarbonyl, and L 1 The precursor of A comprises a precursor that reacts chemically with the precursor of A to form AL 1 Key functional groups.

14. The compound comprising a self-destructing group according to claim 1, wherein L 1 C 1-200 The alkylene group, the middle of which chain optionally contains a divalent or multivalent functional group selected from the group consisting of amide, sulfonamide, amino, ether, carbonyl, triazole, tetrazole, sugar-derived group, sulfonated ester and dendrimer.

15. The compound comprising a self-destructing group according to claim 1, wherein L 1 Including any one selected from the group consisting of: -(CH2) na -;-(CH2CH2O) ma -;-(CH2OCH2) mb -;-(OCH2CH2) mc -;-C(=O)-; or a combination thereof, and R d H or C 1-8 Alkyl, and na and ma to mc are each independently an integer of 0 to 10.

16. The compound comprising a self-destructing group according to claim 1, wherein AL 1 -Selected from the group consisting of: N3-(CH2) n1 -; N3-(CH2CH2O) m1 -(CH2) n2 -;HO-(CH2CH2O) m1 -(CH2) n2 -;H2N-(CH2CH2O) m1 -(CH2) n2 -;H2N-O-(CH2CH2O) m1 -(CH2) n2 -; N3-(CH2CH2O) m2 -(CH2) n3 -NR d1 CO-(CH2) n4 -; R a1 NH-(CH2) n5 -; R b1 OC(=O)-(CH2) n6 -; R c1 C≡C-(CH2OCH2) m3 -CONR d2 -(CH2) n7 -; R a1 , R b1 , R c1 , R c2 and R d1 To R d5 Each independently is H or C 1-8 An alkyl group, and n1 to n15 and m1 to m11 are each independently an integer of 0 to 10.

17. The compound comprising a self-destructing group according to claim 1, wherein The compound represented by Formula 1 is selected from the group consisting of compounds represented by the following formulae: In the above formula, A, L 1 , k, V, R 1 , R 2 , R 3 , R 11 , R 12 and PL is as defined in any one of claims 1 to 16.

18. The compound comprising a self-destructing group according to claim 1, wherein The compound represented by Formula 1 is a compound represented by the following Formula 1-1: [Formula 1-1] In formula 1-1, A has the same meaning as A defined in claim 1; U 1 and U 2 Each has the same meaning as U defined in claim 1, wherein U 1 and U 2 the same as or different from one another; j is 1 to 10; L 11 and L 12 Each having L as defined in claim 1 1 The same meaning, where L 11 and L 12 the same as or different from one another; L 1a and L 1b are each independently selected from a direct bond; And R e H or C 1-8 Alkyl; and q1, q2 and q3 are each independently an integer from 0 to 10, and q4 is an integer from 1 to 10; The condition is that if L 1a for Then q2 is not 0, and if L 1b for Then q3 is not 0.

19. The compound comprising a self-destructing group according to claim 18, wherein In the above formula 1-1 Select from the following structures: wherein q2 and q3 are as defined in claim 18.

20. The linker compound according to claim 18, wherein L 11 and L 12 Each independently selected from: -(CH2) n1 -;-(CH2CH2O) m1 -(CH2) n2 -;-(CH2CH2O) m2 -(CH2) n3 -NR d1 CO-(CH2) n4 -;-(CH2CH2O) m10 -(CH2) n15 -CONR d5 -(CH2) n14 -and R d1 and R d5 Each independently is H or C 1-8 Alkyl, and n1, n2, n3, n4, n8, n14 and n15 and m1, m2, m8 and m10 are each independently an integer of 1 to 8.

21. The compound according to claim 18, wherein The compound represented by formula 1-1 is selected from the group consisting of compounds represented by the following formulae: In the above formula, q1 to q4, U 1 and U 2 As defined in claim 18, R d1 , R d5 and R e Each independently is H or C 1-8 Alkyl, and n1, n3, n4, n8, n14 and n15, m2, m8 and m10 are each independently an integer of 1 to 8.

22. A ligand-drug conjugate represented by the following formula 2, or a pharmaceutically acceptable salt thereof: [Formula 2] In formula 2, E is a ligand or protein with receptor binding properties; A' is a divalent linking group derived from the binding functional group of A; n is a real number from 1 to 10; and A, U, L 1 , k and j are as defined in claim 1.

23. The ligand-drug conjugate according to claim 22, wherein The ligand is selected from the group consisting of peptides, tumor cell-specific peptides, tumor cell-specific aptamers, tumor cell-specific carbohydrates, tumor cell-specific monoclonal or polyclonal antibodies and antibody fragments; and The protein is selected from the group consisting of C 1-20 A group consisting of hydrocarbon groups, oligopeptides, polypeptides, fragments of antigenic polypeptides and artificial antibodies (Repebodies).

24. The ligand-drug conjugate according to claim 23, wherein The antibody is selected from the group consisting of intact polyclonal antibodies, intact monoclonal antibodies, antibody fragments, single-chain Fv (scFv) mutants, multispecific antibodies, bispecific antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising the antigenic determining portion of antibodies, and other modified immunoglobulin molecules comprising an antigen recognition site.

25. The ligand-drug conjugate according to claim 23, wherein The antibody is selected from the group consisting of: muromonab-CD3, abciximab, rituximab, daclizumab, palivizumab, infliximab, trastuzumab (Herceptin), etanercept, basiliximab, gemtuzumab, alemtuzumab, ibritumomab tiuxetan, adalimumab, afacept, omalizumab, efalizumab, tositumomab-I131, cetuximab, bevacizumab, natalizumab, ranibizumab, panitumumab, eculizumab, rilonacept, certolizumab pegol, romiplostim , AMG-531 (lomiperol), CNTO-148 (golimumab), CNTO-1275 (ustekinumab), ABT874 (briakinumab), LEA-29Y (belatacept), belimumab, TACI-Ig (transmembrane active agent and calcium regulator and cyclophilin ligand interactor-immunoglobulin), second-generation anti-CD20, ACZ-885 (canakinumab), tocilizumab, atelizumab, mepolizumab, pertuzumab, Humax CD20 (ofatumumab), tremelimumab (CP-675206), tesimumab, MDX-010 (ipilimumab), IDEC-114 (galiximab), ointuzumab, HuMax EGFR (zalumab), aflibercept (VEGF Trap-Eye), HuMax-CD4 (zalumab), Ala-Ala (hOKT3gamma1), otelizumab (ChAglyCD3; TRX4), catumaxomab, MT-201 (adelimumab), Pregovomab, CH-14.18 (detucilizumab), WXG250 (gemtuximab), AMG-162 (denosumab), AAB-001 (bapinetumab), motavizumab, MEDI524 (motavizumab), ifengumab, Rabikal, third-generation anti-CD20, LY2469298 (Ocatatumumab), and Veltuzumab.

26. The ligand-drug conjugate according to claim 22, wherein E is an antibody, and the E-A' binding structure in Formula 2 includes Wherein * is the remaining part of the antibody.

27. The ligand-drug conjugate according to claim 22, wherein The conjugate represented by Formula 2 is selected from the following conjugates: In the above formula, mAb represents the antibody portion; m5, m6, m9, m10, n9, n10, n13, n14 and n15 are each independently an integer from 1 to 10; R d3 and R d5 Each independently is H or C 1-8 alkyl; Z 1 Selected from NR 3 , O, S and Se heteroatoms, R 3 H or C 1-8 Hydrocarbon; PL is part of the active agent; and n is a real number from 1 to 10.

28. The ligand-drug conjugate according to claim 27, wherein The ligand-drug conjugate represented by Formula 2 is selected from the group consisting of conjugates represented by the following formulae: In the above formula, mAb is a part of an antibody, and n is a real number from 1 to 10.

29. The ligand-drug conjugate according to claim 22, wherein The compound represented by Formula 2 is a compound represented by the following Formula 2-1: [Formula 2-1] In formula 2-1, E is a ligand or protein with receptor binding properties; A' is a divalent linking group derived from the binding functional group of A defined in claim 1; U 1 and U 2 Each has the same meaning as U defined in claim 1, wherein U 1 and U 2 the same as or different from one another; j is 1 to 10; L 11 and L 12 Each having L as defined in claim 1 1 The same meaning, where L 11 and L 12 the same as or different from one another; L 1a and L 1b are each independently selected from a direct bond; And R e H or C 1-8 Alkyl; and q1, q2 and q3 are each independently an integer from 0 to 10, and q4 is an integer from 1 to 10; The condition is that if L 1a for Then q2 is not 0, and if L 1b for Then q3 is not 0; and n is a real number from 1 to 10.

30. The ligand-drug conjugate according to claim 29, wherein The conjugate represented by formula 2-1 is selected from the group consisting of conjugates represented by the following formulae: In the above formula, mAb is the antibody portion; q1 to q4, n, U 1 and U 2 As defined in claim 29; R d1 , R d5 and R e Each independently is H or C 1-8 Alkyl, and n1, n3, n4, n8, n14 and n15, m2, m8 and m10 are each independently an integer of 1 to 8.

31. The ligand-drug conjugate according to claim 29, wherein The conjugate represented by formula 2-1 is selected from the group consisting of conjugates represented by the following formulae: In the above formula, mAb is a part of an antibody, and n is a real number from 1 to 10.

32. A pharmaceutical composition comprising the ligand-drug conjugate according to any one of claims 22 to 31, and a pharmaceutically acceptable carrier or excipient.

33. The pharmaceutical composition according to claim 32, wherein The pharmaceutical composition is used for treating or preventing proliferative diseases, autoimmune diseases or infectious diseases.

34. An imaging composition comprising the ligand-drug conjugate according to any one of claims 22 to 31.

35. A composition for detection comprising the ligand-drug conjugate according to any one of claims 22 to 31.

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