Novel linker compounds and ligand-drug conjugates thereof

Abstract

The present invention provides novel linker compounds and ligand-drug conjugates thereof. The composition can stably deliver an active ingredient to a target site and rapidly release the active ingredient at the target site, so that the efficacy of the active ingredient can be increased and side effects of the active ingredient at positions other than the target site can be prevented.

Description

Technical Field

[0001] The present invention relates to a novel linker compound and a ligand-drug conjugate thereof, and specifically to a novel linker compound 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 diagnosis.

[0003] Meanwhile, since bioactive substances may be toxic to non-target biological materials, technologies have been proposed to suppress drug side effects while allowing drug effects to appear selectively on target cells.

[0004] For example, antibody-drug conjugate (ADC) is a targeted technology that combines drugs or toxins with antibodies that bind to receptors in the body, and then selectively releases the drugs or toxins from target cells to exhibit the desired drug effect. Since 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 the in vivo circulation process, and then separates the drug 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 an important influence 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] As non-cleavable linkers, thioethers are mainly used. In cells, the bond between the drug and the linker does not dissociate, but the bond between the linker and the antibody dissociates, and the drug bound to the linker separates from the antibody. Thiol-maleimide linkers are mainly used, but they have the disadvantages of 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] As linkers with a chemical separation mechanism, linkers composed of disulfide bonds, hydrazone bonds, or oxime bonds are generally used. However, chemically separated linkers may cause the drug to dissociate at a position unrelated to the target site depending on the conditions in the blood or cells, thereby causing toxic side effects.

[0010] In order to solve this problem, linkers that selectively hydrolyze in target cells through enzymatic reactions are being developed. For example, linkers that hydrolyze through enzymatic reactions are not directly connected to the drug, but are connected through a self-immolative group (SIG) between the drug and the linker. After the drug is hydrolyzed through enzymatic reactions, it dissociates through 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 linker compound of Formula 1, Formula 1-1 or Formula B, 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, Formula 1-1 or Formula B, 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. That is to say, all combinations of the above-mentioned 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, a linker compound represented by the following Formula 1, or a pharmaceutically acceptable salt thereof is provided:

[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, L1 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 the binding functional group of A.

[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 a receptor binding property.

[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] PL is an active agent which is bonded to L via a heteroatom selected from N, O and S. 2 or with benzene ring and A-(L 1 ) k - the carbon atom to which it is attached;

[0031] L 2 is a self-eliminating joint, which is selected so that L 2 With benzene ring and A-(L 1 ) k - The cleavage of the bond between the bound carbon atoms promotes L 2 Cleavage of the bond with PL;

[0032] W is an optional substituent on the benzene ring;

[0033] Z 1 and Z 3 One of the following is selected from NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 are each independently CH or N, and -(V) h If present, each independently replaces the H in NH or CH;

[0034] Indicates that A-(L 1 ) k - bonds formed;

[0035] R 3 is H or a C1-C8 hydrocarbon group;

[0036] V is an electron-withdrawing group, an electron-donating group or -CH2-(L 2 ) l -PL-;

[0037] T is a trigger group capable of initiating the release of PL and L2 if present, via a 1,6-elimination reaction upon cleavage;

[0038] L 3 is an optional self-destructive spacer group, which, if present, is cleaved sequentially when T is cleaved;

[0039] X and Y are each independently selected from -O-, -NH- and S; and

[0040] h, i, l, x and y are each independently 0 or 1, and p is an integer from 0 to 2.

[0041] Linker compound of formula 1

[0042] Contains fused ring core unit

[0043] Formula 1(A-(L 1 ) k -U j ) in which U is represented by formula A:

[0044] [Formula A]

[0045]

[0046] 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 or, if L 1 Contains branched or dendritic structures (i.e., if L 1 is a multivalent linking group), then multiple U can be connected with L 1 In this case, with L 1 The number j of U bound may be from 1 to 10. In one embodiment, j is from 1 to 5. In one embodiment, j is 1.

[0047] In the above formula A, Z 1 and Z 3 One of the following is selected from NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 Each is independently CH or N. -(V) hIf present, each independently replaces the H in NH or CH. In this case, R 3 It is H or C1-C8 hydrocarbon group.

[0048] In the above formula A, connected to the benzene ring and (L 2 ) l -PL is bound to the carbon atom Indicates the A-(L 1 ) k -bonds formed.

[0049] In one embodiment, Z 1 and Z 3 One of the following is selected from NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 Each independently represents C or N bonded to V. 3 is H or C1-C8 alkyl. 3 Can be H, C 1- C4 alkyl or C 1- C3 alkyl.

[0050] 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.

[0051] In one embodiment, Z 1 and Z 3 One of the following can be selected from NR 3 , O, S and Se, and the other and Z 2 In this case, any CH can be replaced by V.

[0052] In one embodiment, Z 1 and Z 3 One of the following can be selected from NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 Can be N, and Z 1 and Z 3 The remaining one of them can be CH or C combined with V.

[0053] In one embodiment, when Z1 To Z 3 When any one or more of is CH, V may 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 or Formula 2 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 choose from the group consisting of:

[0056]

[0057] The benzene ring in the fused ring structure of the above formula A may be substituted by an optional substituent, 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 may be selected from the group consisting of: H, C1-C 12 Saturated or unsaturated hydrocarbon group, halogen, halogenated-C1-C8 alkyl, CN, NO2, OH, C1-C8 alkoxy, hydroxy-C1-C8 alkyl, C1-C8 alkoxy-C1-C8 alkyl, SH, C1-C8 alkylthio, mercapto-C1-C8 alkyl, amino, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, amino-C1-C8 alkyl, C1-C8 monoalkylamino-C1-C8 alkyl, C1-C8 dialkylamino- C1-C8 alkyl, carboxyl, C1-C8 alkoxycarbonyl, C1-C8 alkoxycarbonyloxy, carboxyl-C1-C8 alkyl, C1-C8 alkoxycarbonyl-C1-C8 alkyl, carbamoyl, mono-C1-C8 alkylcarbamoyl, di-C1-C8 alkylcarbamoyl, carbamoyl-C1-C8 alkyl, mono-C1-C8 alkylcarbamoyl-C1-C8 alkyl and di-C1-C8 alkylcarbamoyl-C1-C8 alkyl. In Formula 1 and Formula 2, p may be an integer from 0 to 2. In one embodiment, p may be 0.

[0058] The fused ring of formula A contains Z 1 , Z 2 and Z 3 The 5-membered ring may be optionally substituted by V.

[0059] In formula A, V is an electron-withdrawing group, an electron-donating group, -CH2-(L 2 ) l -PL-. h and l are each 0 or 1. For example, V can be an electron withdrawing group. For example, V can be -CH2-(L 2) l -PL-.

[0060] In some embodiments, V can be halogen, CN, NO2, formyl, C1-C8 alkylcarbonyl, carboxyl, C1-C8 alkoxycarbonyl, carboxyl-C1-C8 alkyl, carbamoyl, mono-C1-C8 alkylcarbamoyl, di-C1-C8 alkylcarbamoyl, C1-C8 alkyl, C1-C8 alkenyl, OH, C1-C8 alkoxy, SH, C1-C8 alkylthiol, NH2, mono-C1-C8 alkylamino, di-C1-C8 alkylamino or C6-C8 alkyl. 18 In one embodiment, V may be -CH2-PL or -CH2-L 2 -PL.

[0061] In some embodiments, V can be carboxyl, C1-C8 alkoxycarbonyl, carboxyl-C1-C8 alkyl, NH2, mono-C1-C8 alkylamino, or di-C1-C8 alkylamino. For example, V can be carboxyl, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, carboxymethyl, carboxyethyl, carboxypropyl, amino, aminomethyl, aminoethyl, or aminopropyl.

[0062] In one embodiment, when V is -CH2-(L 2 ) l -PL, L 2 , PL and l may each independently have L associated with Formula A below 2 , PL and l have the same definition. In this case, formula A comprises -(L 2 ) l- Two instances of PL, each instance can be identical or different from each other.

[0063] Self-eliminating joints

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

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

[0066] In one embodiment, L 2 It may be at least one selected from -OC(=O)-, -S(=O)2-, In this case, R 10 To R 12 Each of them may be independently H, C1-C8 alkyl, amino-C1-C8 alkyl, C1-C8 alkyl substituted with mono-(C1-C8 alkyl)amino or di-(C1-C8 alkyl)amino, 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- The C8 alkyl group may be a C1-C6 alkyl group, a C1-C4 alkyl group or a C1-C3 alkyl group. 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. PL can be linked to L via a heteroatom selected from N, O and S in the active agent. 2 or with benzene ring and A-(L 1 ) k - is connected to the carbon atom to which it is bonded. In addition, when V is -CH2-(L 2 ) l When -PL, the heteroatom selected from N, O and S in the active agent is combined with -CH2-(L 2 ) l -linked. In this case, the moiety of formula A can carry two active agent moieties. When T is cleaved by an enzymatic reaction or a chemical reaction, PL is released from the compound of formula 1 by 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 (N, O or S) contained in the drugs, toxins, fluorophores, affinity ligands, diagnostic substances and detection probes can be combined with -CH2-(L 2 ) l -bond. Therefore, as long as it contains a heteroatom capable of forming the above-mentioned bond, or it can further introduce a functional group containing such a heteroatom, it can be used 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 of a primary or secondary amine group, the H of a hydroxyl group, or the H of a carboxyl group is removed, a functional group in which the lone pair of electrons of a nitrogen atom of a tertiary amine group is given, or a functional group connected to a nitrogen atom by an addition reaction of an imine group.

[0070] The drug may be selected from, but is not limited to, the group consisting of: 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 (SCH 66336); sorafenib (BAY43-9006; Bayer Labs.); gefitinib (IRESSA; Astrazeneca); AG1478, AG1571 (SU 5271; Sugen); alkylating agents (e.g., thiotepa or cyclophosphamide); alkyl sulfonates (e.g., busulfan, improsulfan, or zinsulfan); aziridines (e.g., benzodopa, carboquinone, metodepa, or uredepa); ethyleneimine, methylmelamine, hexamethylmelamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphamide, trimethylolmelamine; acetogenins (e.g., bratacin or bratacinone); camptothecins including synthetic mimetics of topotecan; bryostatin; callystatin; CC-1065 (including its acetogenins); dolexin, carzelesin or biezelesin synthetic analogs); cryptophycins (e.g., cryptophycin 1 or cryptophycin 8); dolastatin; duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; panratistatin; sarcodictyin; spongistatin; nitrogen mustards (e.g., chlorambucil, naphthyl mustard, chlorophosphamide, estramustine, ifosfamide amine, nitrogen mustard, nitrogen mustard oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trolofosamide or uracil mustard); nitrosoureas (e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine or ranimnustine); antibiotics (e.g., enediyne antibiotics, calicheamicin selected from calicheamicin gamma and calicheamicin omega, or dynemicins, including dynemicin A); bisphosphonates (e.g., clodronate); emphosphonates (e.g., clodronate); esperamicin, neospermicide chromophore, or related chromoprotein enediyne antibiotic chromophore, aclacinomycins, actinomycins, anramycins, azaserine, bleomycins, cactinomycins, carabicin, carninomycin, carbomycin, chromomycin, dactinomycin, daunorubicin, detorubucin, 6-diazo-5-oxo-L-norleucine, ADRLIMYCIN;doxorubicin (e.g., morpholino doxorubicin, cyanomorpholino doxorubicin, 2-pyrroline-doxorubicin, liposomal doxorubicin, or deoxydoxorubicin), epirubicin, esorubicin, mexilomycin, mitomycin (e.g., mitomycin C, mycophenolic acid, nogamycin, olivetomycin, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptomycin, streptozocin, tuberculocin, ubenimex, zinostatin, or zorubicin); antimetabolites (e.g., 5-fluorouracil (5-FU)); folic acid analogs (e.g., folinic acid, methotrexate, pteropterin, or trimetrexate); purine analogs (e.g., fludarabine, 6-fluorouracil, 5-fluorouracil ... -mercaptopurine, thioether purine, or thioguanine); pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, or floxuridine); androgens (e.g., captestosterone, drostanolone propionate, cyclothiodine, melastosane, or testolactone); antiadrenal agents (e.g., aminoglutethimide, mitotane, or trilostane); folic acid supplements (e.g., folinic acid); aceglucuronolide; aldophosphamide glycosides; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; anthracene; edatraxate; defofamine; demecil; diaziquone; elfornithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansine-like (e.g., maytansine or ansamitocin); trichothecenes (e.g., T-2 toxin, verracurin A, baculosporin A, or anguidine); mitoxantrone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; methylprocarbazine; polysaccharides; razoxane; rhizoxin; siroxan; spirogermanium; tenuazonic acid; triazine quinone; 2,2',2"-trichlorotriethylamine; trichothecenes (particularly, T-2 toxin, verracurin A, baculocin A and anguidine); urethanes; vindesine; dacarbazine; mannitol mustard; dibromomannitol; dibromodulanol; piperobroman; gacytosine; cytarabine ('Ara-C'); cyclophosphamide; thiotepa; taxanes (e.g. Paclitaxel (TAXOL; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANET™ 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; daunomycin; 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) anti-hormonal agents that regulate 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) anti-androgen agents, such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and troxacitabine (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 ribozymes 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 for cell signaling secreted by many cells, which can be signaling molecules widely used for intercellular communication. Cytokines include mononuclear cytokines, lymphocyte factors, 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 may be selected from the group consisting of aminocaproic acid, azathioprine, bromocriptine, chloroquine, chlorambucil, cyclosporine, cyclosporine 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, taxol, 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, exotecan, belotecan, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone , asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, chlorambucil, melphalan, carmustine, lolimus, busulfan, sulfan, dacarba, etoposide, teniposide, topotecan, 9-aminocamptothecin, crestofol, mitomycin C, trimetrexate, mycophenolic acid, thiazofurine, ribavirin, EICAR (5-ethynyl-1-β-D-ribofuranosyl imidazolyl-4-carboxamide), hydroxyurea, deferoxamine, floxuridine, doxifluridine, raltitrexed, cytarabine (arabinoside) C), cytosine arabinoside, fludarabine, tamoxifen, raloxifene, megestrol acetate, goserelin, leuprolide 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, pelocycin, epirubicin, pirarubicin, doxyrubicin, mitoxantrone, verapamil and thapsigargin. The antiviral agent can be selected from the group consisting of: pencicyclovir, valacyclovir, gancicyclovir, foscarnet, ribavirin, iodoxuridine, 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, candidacin, 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, Peru balsam, 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 in living cells or organisms. A toxin can be a small molecule, peptide or protein, which can interact with biological macromolecules (such as enzymes or cell receptors) after contacting or being absorbed by body tissues to cause disease. In addition, "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, bilaterian 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 (EP 1391213, ACR 2008, 41, 98-107), calicheamicin (US 2009105461, Cancer Res. 1993, 53, 3336-3342), daunomycin, doxorubicin, methotrexate, vinblastine, SG2285 (Cancer Res. 2010, 70 (17), 6849-6858), dolastatin, dolastatin analogs auristatin (US563548603), cryptophycin, camptothecin, root toxin derivatives, CC-1065 analogs or derivatives, multicarmycin, enediyne antibiotics, esperamicin, epothilone, 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 may include molecules that can form complexes with target biomolecules. Affinity ligands may be molecules that transmit signals by binding to a predetermined position on a target protein. Affinity ligands may be substrates, inhibitors, stimulants, neurotransmitters, or radioisotopes.

[0079] The term "detection probe" may 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 may include 32P, 35S, fluorescent dyes, electron-dense reagents, enzymes (e.g., enzymes commonly used in ELISA), biotin-streptavidin, digoxigenin, haptens, and proteins for which antiserum or monoclonal antibodies are available, or nucleic acid molecules with sequences complementary to the target. Detection probes are generally capable of generating measurable signals, such as radioactive, chromogenic or fluorescent signals, which can be used to quantify the amount of detectable moieties bound in a sample. Quantification of signals 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 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 (e.g., 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 can 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] In one embodiment, PL may be selected from a moiety represented by the following formula:

[0087]

[0088] Trigger unit

[0089] 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.

[0090] T can be selectively cleaved in vivo by chemical reaction or enzymatic reaction, 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.

[0091] 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.

[0092] 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.

[0093] β-galactoside and β-glucuronide can be connected to the benzene ring via the oxygen atom connected to the C1 position of the β-galactoside part and the β-glucuronide part, and the valine-citrulline derivative and the valine-alanine derivative can be connected to the benzene ring via the nitrogen atom of the valine part, that is, the -β-galactoside, -β-glucuronide, -valine-citrulline derivative and -valine-alanine derivative herein can be a part having the following structure:

[0094]

[0095] R t1 and R t2 It can be C1-C8 alkyl, Ar 1 and Ar2 Can be C5-C 20 In addition, R t3 and R t4 and s. In one embodiment, the arylene group may be phenyl or naphthyl. In one embodiment, the heteroarylene group may contain 1 to 3 heteroatoms (N, O or S). For example, the heteroarylene group may be furanyl, thienyl or pyrrolyl.

[0096] R t5 It can be OH, mono-C1-C8 alkylamino, di-C1-C8 alkylamino or -NH(CH2CH2O) f R t6 , where R t6 It can be H or C1-C4 alkyl, and f can be an integer from 1 to 10.

[0097] 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 cracked under reducing conditions.

[0098] 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.

[0099] Optional self-destruct spacer unit

[0100] 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 that can transfer the electrons generated when T is cleaved to the benzene ring of formula A and PL.

[0101] In one embodiment, -(X) x -L 3 - can be Here, R 8 and R 9 can be independently selected from the group consisting of H, halogen, C1-C8 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.

[0102] In one embodiment, Z 1 and Z 3 One of them can be O, and -(X) x -L 3 - can be

[0103] In formula A of the present invention, i may be 0 or 1. In one embodiment, if i is 0, the self-destructive spacer may not exist, and -(Y)yT may be directly connected to the benzene ring.

[0104] In one embodiment, in formula A of the present invention, i can be 0 (i.e., there is no self-destructive spacer group), and the self-destructive linker L 2 It may be -O(C=O)-, in which case Formula A of the present invention can be represented by the following Formula A-1.

[0105] [Formula A-1]

[0106]

[0107] In formula A-1, Z 1 , Z 2 , Z 3 , V, Y, T, W, PL, h, y and p are as defined above for Formula A.

[0108] Combination unit

[0109] 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.

[0110] 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. 1That 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 (not present) 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 linker, and it should be noted that the binding functional group of the present invention is not limited to the specific functional groups exemplified herein. For example, the binding functional group can be bound to a ligand or protein (E) having receptor binding properties, or a linker precursor, by a click chemistry reaction. In one embodiment, when L in Formula 1 1 When L 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 combined with an additional linker precursor to form an extended linker, which can then bind to a ligand having receptor binding properties through the functional group contained in the additional linker precursor.

[0111] 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 a receptor binding property, and can be, for example, an intermediate 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 should not bind to a ligand having a receptor binding property. The complex can be used for various purposes, such as changing the properties of the active agent (e.g., water solubility), targeting, etc.

[0112] In one embodiment, the binding functional group may include or consist of a functional group selected from the group consisting of halogen, OH, C1-C8 alkoxy, hydroxyamino, COH, C1-C8 alkylcarbonyl, carboxyl, C1-C8 alkoxycarbonyl, toluenesulfonyl, toluenesulfonate, amino, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, NHNH2, N3, haloacetamide, maleimido, succinimidyl, SH, SO3H, C1-C8 alkylsulfonyl, C1-C8 alkoxysulfonyl, 2-pyridyl disulfide, PO3H2, OPO3H2, -N≡C, -NCS, C4-C 10Dienyl, C2-C8 alkenyl, C2-C8 alkynyl, C4-C 10 Cycloalkynyl and C2-C8 alkynylcarbonyl. f Each of A and A can be independently H or C1-C8 alkyl. In one embodiment, A can be maleimide, hydroxyamino, carboxyl, amino, N3, C2-C8 alkynyl or For example, a maleimide group may refer to a nitrogen atom of maleimide and L 1 or U. In one embodiment, the C1-C8 alkyl group may be a C1-C6 alkyl group, a C1-C4 alkyl group or a C1-C3 alkyl group. In one embodiment, the C2-C8 alkynyl group may be an ethynyl group, a propynyl group (propargyl group) or a butynyl group.

[0113] In Formula 1 of the present invention, L 1 L is a linking group connecting A and the part U represented by formula A, and k is 0 or 1. 1 The position of attachment to the moiety U is represented in formula A by express.

[0114] In one embodiment, when k in Formula 1 is 0, A can be directly combined with the moiety U represented by Formula A. In this case, the position where A is connected to the moiety U is in Formula A. express.

[0115] In one embodiment, AL in Formula 1 1 It can be made by the precursor of A and L 1 The precursors of A and L 1 The bonding 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.

[0116] 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 chemical reaction with the 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.

[0117] In one embodiment, L 1 Can be C1-C 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. 200 The alkylene group can be C1-C 150 Alkylene, C1-C 100 Alkylene, C1-C 80 Alkylene, C1-C 60 Alkylene, C1-C 50 Alkylene, C1-C 40 Alkylene, C1-C 30 Alkylene, C1-C 20 Alkylene or C1-C 10 Alkylene. A sugar-derived group refers to any chemical structure formed between a sugar molecule and another group by a covalent bond. 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 on a core. In the field of ligand-drug conjugates, various dendrimer structure linkers 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.

[0118] In one embodiment, when L 1 When it contains multivalent functional groups, that is, when L 1 When it contains 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.

[0119] In one embodiment, L 1 The structure of the -U bond (e.g., carboxyl, aminocarbonyl, amino, etc.) can be specified as L 1 A substituent of the carbon atom at the position to which the U moiety represented by formula A is bonded.

[0120] In one embodiment, L 1 Can include C1-C 10 Alkylene, oxyethylene, amide, triazole ring, tetrazole ring, ether, carbonyl or a combination thereof.

[0121] In one embodiment, L 1 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 It may be H or a C1-C8 alkyl group, and 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.

[0122] In one embodiment, in Formula 1, k may be 1, and AL 1 - may be selected from the group consisting of: R a1 C≡C-(CH2) n1 -; N3-(CH2) n2 -; In this case, R a1 , R b1 , R f1 , R f2 , R i1 and R d1 To R d10 can be independently H or C1-C8 alkyl. c1 , R e1 , R g1 , R h1 and R h2 Each of the above groups may be independently H, C1-C8 alkyl, C1-C8 alkylcarbonyl (e.g., acetyl), C1-C8 alkoxycarbonyl (e.g., tert-butyloxycarbonyl), or C6-C 12 Aryl-C1-C4 alkoxycarbonyl (e.g., benzyloxycarbonyl). n1 to n18 and m1 to m14 may each independently be an integer from 0 to 10. In one embodiment, n1 to n18 and m1 to m14 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 n18 and m1 to m14 may each independently be an integer from 1 to 8, an integer from 1 to 6, or an integer from 1 to 5.

[0123] Exemplary compounds of Formula 1

[0124] In one embodiment, the compound represented by Formula 1 of the present invention may be a compound represented by the following formula.

[0125]

[0126]

[0127] In the above formula, A, L 1 , k, R 3 and PL is as described in relation to Formula 1. In the above formula, V" represents an electron withdrawing group, for example, V" may be a carboxyl group, a carboxyl-C1-C8 alkyl group or a C1-C8 alkoxycarbonyl group, for example, V" may be a carboxyl group, a carboxymethyl group, a carboxyethyl group, a carboxypropyl group, a methoxycarbonyl group, an ethoxycarbonyl group or a propyloxycarbonyl group.

[0128] The structure A-(L 1 ) k -、V”、L 2 and-(Y) y The combination of -T- is only an example, and based on the embodiments described below and the disclosure of the present 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 the present application.

[0129] 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 compounds shown in Table A attached hereto.

[0130] 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 Table B attached.

[0131] DAR4 type compound (Formula 1-1)

[0132] 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 combining two or more active agents can be provided. In one embodiment, the compound represented by Formula 1 of the present invention can be a compound represented by the following Formula 1-1.

[0133] [Formula 1-1]

[0134]

[0135] In Formula 1-1, A has the same meaning as in Formula 1.

[0136] 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 May be the same as or different from each other.

[0137] In formula 1-1, L 11 and L 12 Each has the same L as in Formula 1 1 The same meaning, where L 11 and L 12 May be the same as or different from each other.

[0138] In Formula 1-1, j is 1 to 10. In one embodiment, j is 1 to 5. For example, j may be 1.

[0139] 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 .

[0140] In formula 1-1, L 1a and L 1b may each independently be selected from a direct bond; In this case, R e It can be H or C1-C8 alkyl.

[0141] 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 C1-C 200 The alkylene group, the middle of which chain optionally contains a divalent or multivalent functional group selected from amide, sulfonamide, amino, ether, carbonyl, triazole, tetrazole, sugar-derived group, sulfonated ester and dendrimer, can be directly bonded to the central N atom.

[0142] 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.

[0143] In one embodiment, in the above formula 1-1 You can choose from the following structures: (where q2 and q3 are defined as above).

[0144] In one embodiment, A in the above formula 1-1 may include a functional group selected from the group consisting of halogen, OH, C1-C8 alkoxy, hydroxyamino, COH, C1-C8 alkylcarbonyl, carboxyl, C1-C8 alkoxycarbonyl, toluenesulfonyl, toluenesulfonate, amino, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, NHNH2, N3, haloacetamide, maleimide, succinimide, SH, SO 3H , C1-C8 alkylsulfonyl, C1-C8 alkoxysulfonyl, 2-pyridyl disulfide, PO3H2, OPO3H2, -N≡C, -NCS, C4-C 10 Dienyl, C2-C8 alkenyl, C2-C8 alkynyl, C4-C 10 Cycloalkynyl and C2-C8 alkynylcarbonyl. f Each of them can be independently H or C1-C8 alkyl. For example, in Formula 1-1, A can be maleimide, hydroxyamino, carboxyl, amino, N3, C2-C8 alkynyl or

[0145] In one embodiment, L 11 and L 12 Can be independently C1-C 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. 200 The alkylene group can be C1-C 150 Alkylene, C1-C 100 Alkylene, C1-C 80 Alkylene, C1-C 60 Alkylene, C1-C 50 Alkylene, C1-C 40 Alkylene, C1-C 30 Alkylene, C1-C 20 Alkylene or C1-C 10 Alkylene.

[0146] 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.

[0147] 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 of 1 to 8, an integer of 1 to 6 or an integer of 1 to 5. In addition, R d1 and R d5 Each is independently H or C1-C8 alkyl.

[0148] In one embodiment, the compound represented by Formula 1-1 (wherein j is 1) may be a compound represented by the following formula:

[0149]

[0150] (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 is independently H or C1-C8 alkyl. )

[0151] In one embodiment, the compound represented by Formula 1-1 may be a compound represented by the following formula:

[0152]

[0153]

[0154] In the above formula, q1 to q4, n1, n8 and m8 are as described above with respect to 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. 3 is H or C1-C8 hydrocarbon group. In the above formula, V" represents an electron withdrawing group, for example, V" can be a carboxyl group, a carboxyl-C1-C8 alkyl group or a C1-C8 alkoxycarbonyl group, for example, V" can be a carboxyl group, a carboxymethyl group, a carboxyethyl group, a carboxypropyl group, a methoxycarbonyl group, an ethoxycarbonyl group or a propyloxycarbonyl group. In the above formula, the -O-CO- group in the -O-CO-PL group is an optional self-destructing spacer group, and may not exist (i.e., the -PL group is directly connected to the -CH2- group), and may also be replaced by the functional group described above with respect to the "optional self-destructing spacer group".

[0155] In one embodiment, the compound represented by Formula 1-1 can be selected from the compounds listed in Table C attached hereto.

[0156] Method for preparing the compound of formula 1

[0157] Based on the examples herein and the technical knowledge of ordinary technicians in the field of organic synthesis, the novel linker compounds of Formula 1 of the present invention can be easily prepared by selecting appropriate solvents, starting materials, intermediates, reaction conditions, etc.

[0158] In one embodiment, the compound of formula 1 having a benzothiophene core and a β-galactoside trigger group can be prepared according to the following reaction scheme 1, wherein A-(L 1 ) k - is propagyl (HCCCH2-).

[0159] [Reaction Scheme 1]

[0160]

[0161] In step 1 of reaction scheme 1, a benzothiophene-based starting material in which the para position of the benzene ring is substituted with COH and OH can be prepared, and the starting material is reacted with galactose in which the hydroxyl group is protected by a protecting group (-PG). For example, a protected galactoside group, such as acetylated galactose or acetyl bromide-α-D-galactose, 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 1, 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 1 can be carried out under 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.

[0162] In step 2, a propargyl group may be introduced to the COH connected to the benzene ring. For example, the product of step 1 may be reacted with a propargyl halide (HCCCH2-Hal; Hal is a halogen). At this time, additives such as 1,2-diiodoethane and zinc powder may be used. The reaction in step 2 may be carried out at a temperature of -10°C to 40°C, -10°C to 30°C, 0°C to 40°C, or 0°C to 30°C, accompanied by ultrasonic stirring.

[0163] In step 3, a self-eliminating linker -OCO- can be introduced. The precursor of the self-eliminating linker -OCO- can use bis(4-nitrophenyl) carbonate, 4-nitrophenyl chloroformate, etc. In the reaction of step 3, additives such as DIPEA and pyridine can be used. Step 3 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.

[0164] In step 4, PL may be introduced. For example, PL may 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 4, additives such as HOBt, pyridine, DIPEA, etc. may be used depending on the type of PL. Step 4 may be performed 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.

[0165] In step 5, 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 5 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.

[0166] Each step in reaction scheme 1 can be carried out in a suitable solvent, and the solvent is 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.

[0167] 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.

[0168] Linker compound of formula B

[0169] In one aspect of the present invention, a novel linker compound represented by the following formula B, or a pharmaceutically acceptable salt thereof is provided:

[0170] [Formula B]

[0171]

[0172] In the above formula B, V' is -CH2-(L 2 ) l -PL.

[0173] In the above formula B, L 2 is a self-eliminating linker, which is selected so that the -CH2- of V' is 2 The cleavage of the bond between L 2 Cleavage of the bond with PL.

[0174] In the above formula B, PL is a heteroatom selected from N, O and S and L of V' 2 or -CH2- linked active agents.

[0175] In the above formula B, Z a and Z b One selected from N, NR 3 , O, S and Se, Z a and Z b The other is CH or N, R 3 It is H or C1-C8 hydrocarbon group.

[0176] In the above formula B, T is a trigger group capable of initiating the release of PL and L2, if present, via a 1,6-elimination reaction upon cleavage.

[0177] In the above formula B, L 3 is an optional self-immolative spacer group which, if present, is cleaved sequentially when T is cleaved.

[0178] In the above formula B, X and Y are each independently selected from -O-, -NH- and -S-, and i, l, x and y are each independently 0 or 1, and p is an integer from 0 to 2.

[0179] In the above formula B, L 2 , PL, X, Y, L 3 The specific embodiments of and T are as described above with respect to Formula 1, and therefore, if applicable, the contents described above with respect to Formula 1 may also apply to Formula B.

[0180] In one embodiment, the compound represented by the above formula B can be selected from the group consisting of compounds represented by the following formulae:

[0181]

[0182] The novel linker compound represented by the above formula B is a complex containing an active agent (PL) that should not bind to a ligand having receptor binding properties. The complex can be used for various purposes, such as changing the properties of the active agent (e.g., water solubility), targeting, etc.

[0183] Ligand-drug conjugate represented by formula 2

[0184] In one aspect of the present invention, a ligand-drug conjugate represented by the following Formula 2 is provided:

[0185] [Formula 2]

[0186]

[0187] In Formula 2 of the present invention, E is a ligand or protein having receptor binding properties.

[0188] 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, a portion 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 -NH2 of unnatural amino acids, can be used as a ligand. The binding functional groups of the compound of formula 1 can be mutually bound. 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.

[0189] 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.

[0190] In one embodiment, the protein can be selected from the group consisting of C1-C 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.

[0191] In one embodiment, the antibody can be selected from the group consisting of an intact polyclonal antibody, an intact monoclonal antibody, an antibody fragment, a single-chain Fv (scFv) mutant, a multispecific antibody, a bispecific antibody, a chimeric antibody, a humanized antibody, a human antibody, a fusion protein comprising an antigenic determining portion of an antibody, and a modified immunoglobulin molecule comprising an antigen recognition site.

[0192] 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, rilanacept, 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 (Inotuzumab), HuMax EGFR (zalumab), aflibercept (VEGF Trap-Eye), HuMax-CD4 (zalumab), Ala-Ala (hOKT3gamma1), Otelixizumab (ChAglyCD3; TRX4), Catumaxomab, MT-201 (adelimumab), Pregovomab, CH-14.18 (detucillinumab), WXG250 (gemtuximab), AMG-162 (denosumab), AAB-001 (bapinetumab), motavizumab, MEDI524 (motavizumab), ifengumab, Rabikal, third-generation anti-CD20, LY2469298 (Ocatatumumab), and Veltuzumab.

[0193] In one embodiment, the antibody can be a monoclonal antibody (mAb).

[0194] 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 about the binding functional group (A) of formula 1 may also apply to A'.

[0195] 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 necessary, a functional group for the above binding may be introduced into the antibody, and the type of such functional group and the method of introducing the functional group are known in the art.

[0196] 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, 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 -NH2 of unnatural amino acids. part.

[0197]

[0198] 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.

[0199] 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 , k and j. 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.

[0200] In one embodiment, the compound represented by the above formula 2 may be selected from the group consisting of compounds represented by the following formulae:

[0201]

[0202] In the above formula, mAb represents a portion of an antibody. In the above formula, m8 and n8 may each independently be an integer from 1 to 10. In one embodiment, m8 and n8 may 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. In the above formula, Z 1 Selected from NR 3 , O, S and Se heteroatoms. 3is H or C1-C8 hydrocarbon group. In the above formula, PL has the same meaning as PL in Formula 1. V" represents an electron withdrawing group, for example, V" can be a carboxyl group, a carboxyl-C1-C8 alkyl group or a C1-C8 alkoxycarbonyl group, for example, V" can be a carboxyl group, a carboxymethyl group, a carboxyethyl group, a carboxypropyl group, a methoxycarbonyl group, an ethoxycarbonyl group or a propyloxycarbonyl group. 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-, Substitution (in the above formula, R 10 To R 12 Each is independently H, C1-C8 alkyl, amino-C1-C8 alkyl, C1-C8 alkyl substituted with mono-(C1-C8 alkyl) or di-(C1-C8 alkyl)amino, or -(CH2CH2O) g R 13 , R 13 is H or C1-C4 alkyl, and g is an integer from 1 to 10).

[0203] 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.

[0204] In one embodiment, the conjugate represented by Formula 2 may be selected from the group consisting of conjugates represented by the following formulae:

[0205]

[0206]

[0207] 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.

[0208] Exemplary DAR2-type ligand-drug conjugates

[0209] 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. In one embodiment, n can be a real number of 1 to 8, a real number of 1 to 6, a real number of 1 to 4, or a real number of 1 to 2.

[0210] DAR4-type ligand-drug conjugate (Formula 2-1)

[0211] 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 1When 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:

[0212] [Formula 2-1]

[0213]

[0214] 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 contents described in Formula 2 are also applicable to Formula 2-1.

[0215] 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 or different from each other; in addition, L 11 and L 12 With the same L as in Formula 1 1 The same meaning, where L 11 and L 12 May be the same as or different from each other.

[0216] In Formula 2-1, j is 1 to 10. In one embodiment, j is 1 to 5. For example, j is 1.

[0217] 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 respectively. 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 respectively.

[0218] In formula 2-1, L 1a and L 1b may each independently be selected from a direct bond; In this case, R e It can be H or C1-C8 alkyl.

[0219] 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.

[0220] 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.

[0221] About L 1a and L 1b , q1, q2, q3 and q4, the contents described in Formula 1-1 can be similarly applied to Formula 2-1 respectively.

[0222] 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.

[0223] In one embodiment, the ligand-drug conjugate represented by Formula 2-1 may be a conjugate represented by the following formula:

[0224]

[0225] (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. )

[0226] In one embodiment, the ligand-drug conjugate represented by Formula 2-1 may be a conjugate represented by the following formula:

[0227]

[0228]

[0229] In the above formula, mAb is part of an antibody. In the above formula, q1 to q4, n1, n8 and m8 are as described above with respect to 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. 3is H or C1-C8 hydrocarbon group. In the above formula, V" represents an electron withdrawing group, for example, V" can be a carboxyl group, a carboxyl-C1-C8 alkyl group or a C1-C8 alkoxycarbonyl group, for example, V" can be a carboxyl group, a carboxylmethyl group, a carboxyethyl group, a carboxypropyl group, a methoxycarbonyl group, an ethoxycarbonyl group or a propyloxycarbonyl group. In the above formula, the -O-CO- group in the -O-CO-PL group is an optional self-destructing spacer group, which may not exist (i.e., the -PL group is directly connected to the -CH2- group), or it may be replaced by the functional group described above with respect to the "optional self-destructing spacer group".

[0230] 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.

[0231] definition

[0232] 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. The hydrocarbyl may include an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, etc. Non-limiting examples of the hydrocarbyl may include methyl, ethyl, propyl, butyl, vinyl, propenyl, butenyl, ethynyl, propynyl, butynyl, etc.

[0233] 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-C8 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.

[0234] The term "alkenyl" includes straight chain 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 and the like.

[0235] The term "alkynyl" refers to a straight or branched hydrocarbon chain having at least one triple bond. Alkynyl is preferably a straight or branched chain having 2 to 8 carbon atoms, and 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, etc.

[0236] The term "alkoxy" refers to an alkyl group bonded 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. An alkoxy group may be a methoxy group, an ethoxy group, or a propoxy group.

[0237] 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, etc.

[0238] 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, etc.

[0239] 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 by heteroatoms (e.g., N).

[0240] The term "diene group" refers to an unsaturated branched or unbranched C4-C 10 Aliphatic substituents. Examples include 2,4-pentadienyl, 2,4-hexadienyl, 4-methyl-2,4-pentadienyl, and the like, but are not limited thereto.

[0241] The term "halogen" atom refers to an atom belonging to Group 17 of the periodic table. Halogen atoms include fluorine, chlorine, bromine, iodine, and the like.

[0242] The term "haloalkyl" refers to an alkyl group substituted with one or more halogen atoms.

[0243] The term "hydroxy" refers to an OH functional group (hydroxyl group).

[0244] The term "thiol" refers to a SH functional group.

[0245] The term "cyano" is CN and refers to a functional group consisting of a triple bond between a carbon atom and a nitrogen atom.

[0246] The term "oxo" refers to =0 and "substituted with oxo" means that a carbon atom has a =0 substituent of the form -C(=O)-.

[0247] The term "nitro" refers to NO2.

[0248] The term "amino" refers to -NH2.

[0249] 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 of the above-mentioned alkyl groups, including monoalkylamino and dialkylamino, and the two alkyl groups in the dialkylamino group may be the same or different. Specifically, a mono-C1-C8 alkylamino group may be a group in which one hydrogen atom of the amino group (-NH2) is replaced by a C1-C8 alkyl group, and a di-C1-C8 alkylamino group may be a group in which two hydrogen atoms of the amino group (-NH2) are replaced by the same or different C1-C8 alkyl groups. For example, a mono-C1-C8 alkylamino group (-NH(C1-C8 alkyl)) may include methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, sec-butylamino, tert-butylamino, pentylamino, hexylamino, etc. The di-C1-C8 alkylamino group (-N(C1-C8 alkyl)2) may include, for example, dimethylamino, diethylamino, dipropylamino, methylethylamino, methylpropylamino, methylisopropylamino, methylbutylamino, methylisobutylamino, ethylpropylamino, ethylisopropylamino, ethylisobutylamino, isopropylisobutylamino, methylhexylamino, ethylhexylamino, and the like.

[0250] The term "carboxyl" refers to -COOH.

[0251] The term "carbamyl" refers to -CONH2.

[0252] The terms "N-mono-C1-C8 alkylcarbamoyl" and "N,N-di-C1-C8 alkylcarbamoyl" refer to a carbamoyl group (-CONH2) in which one or two hydrogen atoms bonded to the nitrogen atom are replaced by a C1-C8 alkyl group. The two C1-C8 alkyl groups in the N,N-di-C1-C8 alkylcarbamoyl group may be the same or different.

[0253] 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.

[0254] 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.

[0255] The terms "cyano-C1-C8 alkyl", "halo-C1-C8 alkyl", "hydroxy-C1-C8 alkyl", "C1-C8 alkoxy-C1-C8 alkyl", "C1-C8 alkoxy-C1-C8 alkoxy-C1-C8 alkyl", "carboxy-C1-C8 alkyl", "amino-C1-C8 alkyl", "carbamoyl-C1-C8 alkyl", "N-mono-C1-C8 alkylcarbamoyl-C1-C8 alkyl" and "N,N-di-C1-C8 alkylcarbamoyl-C1-C8 alkyl" refer to C1-C8 alkyl substituted at the terminal or in the middle with cyano, halogen, hydroxy, alkoxy, carboxyl, amino, carbamoyl, N-mono-C1-C8 alkylcarbamoyl and N,N-di-C1-C8 alkylcarbamoyl, respectively.

[0256] The term "glycosidic group" refers to a functional group formed by a condensation reaction of a sugar molecule.

[0257] 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.

[0258] 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.

[0259] The term "heterocyclyloxy" refers to a functional group in which an oxygen atom is directly attached to a heterocyclic ring.

[0260] 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. The heteroaryl group can contain, for example, 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 or 2 heteroatoms. The heteroaryl group can contain 5 to 10 ring elements, 5 to 7 ring elements, or 5 or 6 ring elements. The heteroaryl group can be a 5 to 6-membered heteroaryl group containing one or two N, O or S. The heteroaryl group can be a monocyclic group, a bicyclic group, or a tricyclic group. The bicyclic group can be a spirocyclic group, a bridged ring group, and a condensed ring group.

[0261] 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 ... 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, etc.

[0262] 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.

[0263] The term "moiety" as used herein refers to the portion of the compound of the parent compound that corresponds to the moiety when the parent compound is combined with the compound of Formula 1 or the conjugate of Formula 2. Even when a portion of the entire compound is referred to herein as a compound or active agent, it should be understood from the context that it refers to the "portion" of the compound or active agent.

[0264] In the present invention, the "precursor" refers to a compound that is a reactant that ultimately provides a desired part through a chemical reaction or the like.

[0265] In the present invention, "linker precursor" refers to a compound that forms a desired linker structure or a part thereof by chemical reaction. For example, as PEG linkers, various substances such as hydroxyl-PEG linkers, alkynyl-PEG linkers, bromine-PEG linkers, DBCO--PEG linkers, azido-PEG linkers, amino-PEG linkers, and maleimide-PEG linkers are known. 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 field. For another example, as a linker having a DBCO (dibenzocyclooctyne) group, there are known 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 other substances. The linker precursor of the present invention includes various linkers known in the ligand-conjugate field, and is not limited to the linker structure exemplified herein. Available linker precursors, preparation methods, reaction conditions, etc. are well known in the relevant field.

[0266] The term "dendrimer" refers to an ordered three-dimensional molecular structure having branched units centered around a core.

[0267] 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).

[0268] In one embodiment, when β-galactose is cleaved by β-galactosidase, the compound of Formula 1 having a benzothiophene core and a β-galactoside trigger group can release PL- or PL-H through a 1,6-elimination reaction.

[0269] [Reaction Scheme A]

[0270]

[0271] 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.

[0272] [Reaction Scheme B]

[0273]

[0274] In addition, when the compound of Formula 1 contains a self-eliminating linker (eg, -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.

[0275] [Reaction Scheme C]

[0276]

[0277] In one embodiment, when the compound of formula B containing a benzothiophene core and a β-galactoside trigger group comprises a -OCO- self-immolative linker (L 2 ), β-galactose and CO2 can separate to release PL - or PL-H, for example, occurs via a 1,6-elimination reaction as shown in Reaction Scheme D below.

[0278] [Reaction Scheme D]

[0279]

[0280] 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 under mild conditions such as room temperature and normal pressure using modular reactants. 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 olefins and azides, reverse demand Diels-Alder reactions of olefins and tetrazines, photo-click reactions of olefins 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-cyclooctenes, nitrones, nitrile oxides, azides, conjugated dienes, dienophiles and cycloalkynes such as cyclooctynes, cyclononynes, dibenzocyclooctynes ​​(DIBO), BARAC (bisaryl azacyclooctynes ​​ketones), ALO (aryl octynes), DIFO (difluorinated cyclooctynes), MOFO (monofluorinated), DIBAC (dibenzoazacyclooctynes) and DIMAC (dimethoxyazacyclooctynes), but are not limited thereto. For example, click chemistry functional groups may include acetylene, trans-cyclooctynes, cyclooctynes, diaryl cyclooctynes, methyl ester phosphine, norbornene, tetrazine, methylcyclopropene, azetidine, cyanide, azide, dibenzocyclooctynes, etc.

[0281] Medical uses and pharmaceutical compositions

[0282] The ligand-drug conjugate of the compound of formula 1 or formula 2 of the present invention has excellent stability in various blood environments. In other words, the active agent can maintain a stable binding state in the plasma environment of mice, rats, dogs and humans, thereby minimizing blood toxicity.

[0283] In addition, the ligand-drug conjugate 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 compound of the present invention can quickly dissociate and release the active agent by reacting with enzymes such as β-galactosidase and glucuronidase under pH 4 to 5, so the compound of the present invention can effectively release the active agent in the target cell intracellular environment (such as tumor lysosome).

[0284] In one aspect of the present invention, a pharmaceutical composition is provided, comprising a linker compound of Formula 1, Formula 1-1 or Formula B 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.

[0285] In another aspect of the present invention, an imaging composition or a detection composition is provided, which comprises a linker compound of Formula 1, Formula 1-1 or Formula B or a ligand-drug conjugate of Formula 2 or Formula 2-1 or a pharmaceutically acceptable salt thereof.

[0286] The linker compound of Formula 1, Formula 1-1 or Formula B and the ligand-drug conjugate of Formula 2 or Formula 2-1 of the present invention may be mixed with a solvent and provided as a composition.

[0287] The composition can be prepared as a liquid solution or suspension in an injectable form. In addition, the composition can be prepared in a solid form suitable for injection, as an emulsion or a polypeptide encapsulated in a liposome. The compound of the present invention or the ligand-drug conjugate 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.

[0288] The composition may contain a diluent, such as water, saline, glycerol, ethanol, etc. Auxiliary substances may be added to the composition, such as a wetting agent or emulsifier, a pH buffer, etc. The composition may be administered parenterally by injection, or may be administered by subcutaneous or intramuscular injection. Additional formulations may be provided, such as suppositories or oral formulations. Oral compositions may be provided as solutions, suspensions, tablets, pills, capsules, or sustained-release formulations.

[0289] The composition can be administered in a manner compatible with the dosage form. The composition comprises a therapeutically effective amount of the compound of the present invention or the 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 according to the type of active agent contained in the compound of the present invention or the ligand-drug conjugate and / or the type of ligand or protein bound 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.

[0290] 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.

[0291] 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., virus, bacteria, fungus, parasite, etc.). These methods include administering to a mammal a therapeutic or preventive 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.

[0292] 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).

[0293] As used herein, the term "pharmaceutically acceptable salt" includes organic salts and inorganic salts. Examples thereof 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, sugar acid salt, 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.

[0294] 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.

[0295] Effects of the Invention

[0296] The compound or ligand-drug conjugate 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 sites.

[0297] 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 a tumor microenvironment. BRIEF DESCRIPTION OF THE DRAWINGS

[0298] Figures 1 to 3 The results of measuring the enzymatic cleavage rates of compounds A-12, A-16, and A-36 using Escherichia coli β-galactosidase or Escherichia coli β-glucuronidase are shown respectively.

[0299] Figure 4 The results of the measurements of chemical stability and plasma stability of Compound A-36 are shown.

[0300] Figure 5 and Figure 6 The results of in vivo activity analysis of ADC-1 to ADC-4 and ADC-10 to ADC-13 are shown.

[0301] Figure 7 Shown are the results of in vivo pharmacokinetic studies of ADC-3 in rats. DETAILED DESCRIPTION

[0302] 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.

[0303] 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:

[0304] Ac:Acetyl

[0305] AcOH: acetic acid

[0306] EA: Ethyl acetate

[0307] MC: dichloromethane

[0308] DMF: dimethylformamide

[0309] EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0310] EDC: N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

[0311] HOBt: 1-Hydroxybenzotriazole hydrate

[0312] ACN: Acetonitrile

[0313] THF: Tetrahydrofuran

[0314] DCC: N,N'-dicyclohexylcarbodiimide

[0315] DMAP: 4-dimethylaminopyridine

[0316] NHS: N-hydroxysuccinimide

[0317] DIPEA: diisopropylethylamine

[0318] TEA: triethylamine

[0319] Boc: tert-butyloxycarbonyl

[0320] LAH: Lithium Aluminum Hydride

[0321] TFA: trifluoroacetic acid

[0322] AgOTf: Silver trifluoromethanesulfonate

[0323] KO t Bu: Potassium tert-butoxide

[0324] MMAF-OMe: monomethyl auristatin F methyl ester

[0325] PPTS: Pyridinium p-toluenesulfonate

[0326] TBAI: Tetrabutylammonium iodide

[0327] DIBAL-H: Diisobutylaluminum hydride

[0328] TEMPO: 2,2,6,6-tetramethylpiperidinyl 1-oxyl, 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical

[0329] MMAE: Monomethyl auristatin E

[0330] NCS: N-chlorosuccinimide

[0331] Preparation Example 1: Preparation of Linker P-1

[0332]

[0333] Compound P-1a (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. tBu (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 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), filtered with diatomaceous earth, and extracted by adding distilled water (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 obtain connector P-1 (820 mg, 66%).

[0334] 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).

[0335] Preparation Example 2: Preparation of Linker P-2

[0336]

[0337] Compound P-2a (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, and then 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 added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, EA (300 mL), distilled water (250 mL) and 2N-hydrochloric acid aqueous solution (50 mL) were added, and the organic layer was extracted 5 times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain the linker P-2 (2.32 g, quantitatively obtained). The obtained linker P-2 was used for the next reaction without additional purification process.

[0338] 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).

[0339] Preparation Example 3: Preparation of Linker P-3

[0340]

[0341] Step 1: Preparation of compound P-3b

[0342] Compound P-3a (tetraethylene glycol, Daejung Chemicals & Metals, CAS No.: 112-60-7, 10 g, 51.49 mmol) was dissolved in MC (200 mL) at 0 ° C under a nitrogen atmosphere, and then 4-methylbenzenesulfonyl chloride (24.54 g, 128.72 mmol) and potassium hydroxide (8.67 g, 154.46 mmol) were added, and the mixture was stirred at room temperature for 15 hours. After the reaction was completed, MC (100 mL) and distilled water (200 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 residue was subjected to column chromatography to obtain compound P-3b (18.98 g, 73.3%) in the form of a colorless oil.

[0343] 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); MS m / z:503[M+H] + .

[0344] Step 2: Preparation of compound P-3c

[0345] Compound P-3b (18.98 g, 37.76 mmol) was dissolved in DMF (95 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (7.36 g, 113.36 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 saturated sodium bicarbonate aqueous solution (200 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 residue was subjected to column chromatography to obtain compound P-3c (8.96 g, 97.1%) in the form of a colorless liquid.

[0346] 1 H-NMR (400MHz, CDCl3) δ3.69–3.66(m,12H),3.40–3.37(m,4H); MS m / z:267[M+Na] + .

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

[0348] Compound P-3c (4.17 g, 17.06 mmol) was dissolved in EA (32 mL), ether (32 mL) and 5% aqueous hydrochloric acid solution (64 mL) at 0°C under a nitrogen atmosphere, and then triphenylphosphine (4.47 g, 17.06 mmol) was added, 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, MC (100 mL) was added to the aqueous layer, and the aqueous layer was washed three times. The obtained aqueous layer was concentrated under reduced pressure to obtain a colorless liquid linker P-3 (4.12 g, 95%).

[0349] 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).

[0350] Preparation Example 4: Preparation of Linker P-4

[0351]

[0352] Step 1: Preparation of compound P-4b

[0353] Compound P-3a (tetraethylene glycol, Daejung Chemicals & Metals, 10 g, 51.49 mmol) was dissolved in MC (200 mL) at 0 ° C under a nitrogen atmosphere, and then 4-methylbenzenesulfonyl chloride (3.27 g, 17.16 mmol) and potassium hydroxide (963 mg, 17.16 mmol) were added, and the mixture was stirred at room temperature for 15 hours. After the reaction was completed, MC (100 mL) and distilled water (200 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 residue was subjected to column chromatography to obtain compound P-4a (2.46 g, 41.1%) in the form of a colorless oil.

[0354] 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); MS m / z:349[M+H] + ,371[M+Na] + .

[0355] Step 2: Preparation of linker P-4

[0356] Compound P-4a (2.46 g, 7.05 mmol) was dissolved in DMF (15 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (690 mg, 10.58 mmol) was added, and the mixture was stirred at 60 ° C for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, EA (50 mL) and distilled water (100 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 residue was subjected to column chromatography to obtain compound P-4 (1.37 g, 88.4%) in the form of a colorless oil.

[0357] 1 H-NMR (400MHz, CDCl3) δ3.73–3.66(m,12H),3.62–3.60(m,2H),3.41–3.39(m,2H),2.42(t,J=6.0Hz,1H); MS m / z:220[M+H] + ,242[M+Na] + .

[0358] Preparation Example 5: Preparation of Linker P-5

[0359]

[0360] Step 1: Preparation of compound P-5

[0361] Compound P-3a (tetraethylene glycol, Daejung Chemicals & Metals, 5 g, 25.74 mmol) was dissolved in THF (60 mL) at room temperature under a nitrogen atmosphere, and then sodium hydride (NaH 60% dispersion in mineral oil, 16.5 mg, 0.41 mmol) was 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 the reaction was completed, saturated sodium chloride aqueous solution (20 mL) was added to terminate the reaction, and the organic solvent was removed by concentrating under reduced pressure. MC (60 mL) was added to the remaining sodium chloride aqueous solution, the organic layer was extracted, and the resulting organic layer was then dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was subjected to column chromatography to obtain compound P-5a (1.39 g, 42%) in the form of a light pink oil.

[0362] 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]+ .

[0363] Step 2: Preparation of compound P-5b

[0364] Compound P-5a (1.39 g, 4.31 mmol) was dissolved in ACN (7.5 mL) at 0 ° C under a nitrogen atmosphere, and then pyridine (4 mL) was slowly added. Then, 4-methylbenzenesulfonyl chloride (p-toluenesulfonyl chloride, 1.1 g, 5.77 mmol) solution dissolved in ACN (10.5 mL) was slowly added over 30 minutes, and the mixture was stirred at room temperature for 6.5 hours. After the reaction was completed, 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 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-5b (1.78 g, 86%) in the form of a colorless oil.

[0365] 1 H-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] + .

[0366] Step 3: Preparation of compound P-5c

[0367] Compound P-5b (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 obtained organic layer was washed three times by adding a saturated sodium chloride aqueous solution (200 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained compound P-5c (1.25 g, 96%) in the form of a colorless oil by column chromatography.

[0368] 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] + .

[0369] Step 4: Preparation of linker P-5

[0370] Compound P-5c (80.5 mg, 0.232 mmol) was dissolved in MC (2.5 mL) at 0°C under a nitrogen atmosphere, and then TFA (2.5 mL) was slowly added, and the mixture was stirred at room temperature for 1.5 hours. After the reaction was completed, the mixture was diluted by adding MC (20 mL) and concentrated under reduced pressure to obtain linker P-5 (67 mg, 99%) in the form of an oil.

[0371] 1 H-NMR (400MHz, CDCl3) δ7.13(brs,1H),3.78(t,J=6.0Hz,2H),3.75–3.60(m,14H),3.40(t,J=4.8Hz,2H),2.64(t,J=6.0Hz,2H); EI-MS m / z:314[M+Na] + .

[0372] Preparation Example 6: Preparation of Linker P-6

[0373]

[0374] Step 1: Preparation of compound P-6a

[0375] Compound P-4 (648 mg, 2.96 mmol) was dissolved in MC (11.5 mL) at 0 ° C under a nitrogen atmosphere, and then 4-methylbenzenesulfonyl chloride (1.13 g, 5.91 mmol) and potassium hydroxide (332 mg, 5.91 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, MC (50 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 residue was subjected to column chromatography to obtain compound P-6a (963 mg, 87.5%) in the form of a colorless oil.

[0376] 1 MS m / z:374[M+H] + ,396[M+Na] + .

[0377] Step 2: Preparation of linker P-6

[0378] Compound P-6a (487.2 mg, 1.30 mmol) was dissolved in ACN (20 mL) at room temperature under a nitrogen atmosphere, and then N-Boc-hydroxylamine (Merck, CAS No.: 36016-38-3, 208 mg, 1.56 mmol) and DBU (0.39 mL, 2.60 mmol) were added in sequence, and the mixture was stirred at 80 ° C for 16 hours. 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 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 obtained residue was subjected to column chromatography to obtain the connector P-6 (83.0 mg, 19.0%) in the form of a colorless liquid.

[0379] 1 H-NMR (400MHz, CDCl3) δ7.53(brs,1H),4.02(t,J=4.8Hz,2H),3.72(t,J=4.8Hz,2H),3.71–3.65(m,10H),3.39(t,J=4.8Hz,2H),1.48(s,9H); EI-MS m / z:357[M+Na] + ,235[M-BOC] + .

[0380] Preparation Example 7: Preparation of Compound PL-1

[0381]

[0382] Step 1: Preparation of compound PL-1b

[0383] 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 the reaction solution at 0°C under a nitrogen atmosphere, and 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 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 PL-1b (3.7 g, 70%).

[0384] 1H-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).

[0385] Step 2: Preparation of compound PL-1d

[0386] 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 by adding 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.

[0387] 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).

[0388] Step 1: Preparation of compound PL-1

[0389] The 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 for 2 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was diluted with EA (100 mL), filtered with a diatomaceous earth filter, and the solution was concentrated under reduced pressure to obtain a colorless oil compound PL-1 (264.3 mg, 99%).

[0390] 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).

[0391] Preparation Example 8: Preparation of Core C-1

[0392]

[0393] Step 1: Preparation of compound C-1b

[0394] 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, and then 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) were slowly added. The mixture was stirred at -30 ° C for 3 hours, and then 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 to obtain compound C-1b (7.74 g, 97%). The obtained compound C-1b was used for the next step without additional purification.

[0395] Step 2: Preparation of compound C-1c

[0396] Compound C-1b (7.74 g, 25.02 mmol) was dissolved in xylene (200 mL) at room temperature under nitrogen atmosphere, and then the mixture was stirred at 150°C for 3 hours. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound C-1c (510.8 mg, 22.7%).

[0397] 1 H-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).

[0398] Step 3: Preparation of compound C-1d

[0399] Compound C-1c (1.14 g, 4.05 mmol) was dissolved in DMF (35 mL) at room temperature under a nitrogen atmosphere, and potassium carbonate (1.68 g, 12.15 mmol) and iodomethane (0.65 mL, 10.53 mmol) were added in sequence, and the mixture was stirred for 12 hours. After the reaction was completed, EA (100 mL) and distilled water (100 mL) were added to extract the organic layer. Distilled water (300 mL) was added to extract the obtained organic layer, and the 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-1d (912.7 mg, 76%).

[0400] 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).

[0401] Step 4: Preparation of compound C-1e

[0402] C-1d (800 mg, 2.70 mmol) was dissolved in MC (40 mL) at -50 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (735 μL, 8.10 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 8.1 mL, 8.1 mmol) were slowly added in sequence, and the mixture was stirred for 2 hours. 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 C-1e (392 mg, 44.8%).

[0403] 1 H-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).

[0404] Step 5: Preparation of Core C-1

[0405] 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.

[0406] 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).

[0407] Preparation Example 9: Preparation of Core C-2

[0408]

[0409] Step 1: Preparation of compound C-2b

[0410] Compound C-2a (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 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 and removed with ether (10 mL), and 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-2b (460 mg, 61.6%).

[0411] 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).

[0412] Step 2: Preparation of Core C-2

[0413] Compound C-2b (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 0.4 N-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, and 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 obtain core C-2 (210 mg, 58%).

[0414] 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).

[0415] Preparation Example 10: Preparation of Core C-3

[0416]

[0417] Step 1: Preparation of compound C-3b

[0418] Compound C-3a (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 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-3b (1.87 g, 99%) in the form of a light orange oil.

[0419] 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).

[0420] Step 2: Preparation of compound C-3c

[0421] Compound C-3b (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, and then n-butyl lithium 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 for another 30 minutes at the same temperature. Distilled water (300 mL) was added to the mixture at the same temperature to terminate the reaction, and then 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 residue obtained by column chromatography gave compound C-3c (1.36 g, 65%) in the form of a light yellow oil.

[0422] 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).

[0423] Step 3: Preparation of Core C-3

[0424] Compound C-3c (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, and EA (100 mL) and distilled water (100 mL) were added at room temperature 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 core C-3 (174 mg, 87.4%) in the form of a light yellow-brown solid.

[0425] 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).

[0426] Preparation Example 11: Preparation of Core C-4

[0427]

[0428] Step 1: Preparation of compound C-4a

[0429] The compound C-3c (1.36 g, 5.09 mmol) prepared in step 2 of Preparation Example 10 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 obtained residue was subjected to column chromatography to obtain compound C-4a (1.43 g, 94%) in the form of an ivory solid.

[0430] 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).

[0431] Step 2: Preparation of compound C-4b

[0432] Compound C-4a (240 mg, 0.80 mmol) was dissolved in MC (15 mL) at -78 ° C under nitrogen protection, 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 obtain compound C-4b (206.7 mg, 79%).

[0433] 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).

[0434] Step 3: Preparation of Core C-4

[0435] Compound C-4b (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 warming 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-4 (32.8 mg, 55%) in the form of an ivory solid.

[0436] 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).

[0437] Preparation Example 12: Preparation of Core C-5

[0438]

[0439] Step 1: Preparation of compound C-5a

[0440] The compound C-3c (2.1 g, 7.82 mmol) prepared in step 2 of Preparation Example 10 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-5a (2.5 g, 95%) in the form of a white solid.

[0441] 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).

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

[0443] Compound C-5a (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 the reaction was completed, the solution was filtered through diatomaceous earth and concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound C-5b (4.3 g, 91%) in the form of a white solid.

[0444] 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).

[0445] Step 3: Preparation of compound C-5c

[0446] Compound C-5b (485 mg, 1.42 mmol) was dissolved in MC (30 mL) at -78 °C under a nitrogen atmosphere, and then 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) 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 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-5c (290 mg, 55%).

[0447] 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).

[0448] Step 4: Preparation of Core C-5

[0449] Compound C-5c (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 then 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 with 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-5 (150 mg, 66%) in the form of a white solid.

[0450] 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).

[0451] Preparation Example 13: Preparation of Core C-6

[0452]

[0453] Step 1: Preparation of compound C-6b

[0454] Compound C-6a (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, and then 5-hexenoic acid methyl ester (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 added in sequence, and the mixture was stirred at room temperature for 1 hour. The mixture was stirred at 60°C for 16 hours, and then EA (100 mL) and 2N-hydrochloric acid aqueous solution (100 mL) were added to extract the organic layer, and 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-6b (340 mg, 34.2%).

[0455] 1H-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).

[0456] Step 2: Preparation of compound C-6c

[0457] Compound C-6b (690 mg, 2.94 mmol) was dissolved in ACN (20 mL) at 0 ° C under a nitrogen atmosphere, and then potassium carbonate (1.01 g, 7.36 mmol) and benzyl bromide (420 μL, 3.53 mmol) were slowly added, and the mixture was stirred at 0 ° C for 30 minutes, and then the mixture was stirred at room temperature for 16 hours. After the reaction was completed, EA (300 mL) and distilled water (300 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-6c (894 mg, 93%) in the form of a yellow oil.

[0458] 1 H-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).

[0459] Step 3: Preparation of compound C-6d

[0460] Compound C-6c (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 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 residue was subjected to column chromatography to obtain compound C-6d (511 mg, 53%) in the form of a yellow oil.

[0461] 1H-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).

[0462] Step 4: Preparation of Core C-6

[0463] Compound C-6d (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-6 (200.2 mg, 52%) in the form of a white solid.

[0464] 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).

[0465] Preparation Example 14: Preparation of Core C-7

[0466]

[0467] Step 1: Preparation of compound C-7b

[0468] Compound C-7a (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-7b (6.6 g, 54%) in the form of a yellow solid.

[0469] 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).

[0470] Step 2-1: Preparation of compound C-7c

[0471] Compound C-7b (1.0 g, 4.99 mmol) was dissolved in ACN (25 mL) at room temperature under a nitrogen atmosphere, and then chloromethyl methyl ether (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 obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained compound C-7c (908 mg, 74%) in the form of a white solid was obtained by column chromatography.

[0472] 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).

[0473] Step 3-1: Preparation of compound C-7d

[0474] Compound C-7c (900 mg, 3.68 mmol) was added to THF (30 mL) at room temperature under a nitrogen atmosphere, the mixture was cooled to -40 ° C, and n-butyl lithium solution (2.5 M n-BuLi in hexane, 1.76 mL, 4.42 mmol) was slowly added dropwise. The mixture was warmed to -20 ° C, DMF (0.85 mL) was slowly added dropwise, slowly warmed 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 with EA (30 mL), 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 obtain compound C-7d (300 mg, 30%) in the form of a white solid.

[0475] 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).

[0476] Step 4-1: Preparation of compound C-7g (Method 1)

[0477] Compound C-7d (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 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-7g (150 mg, 60%) in the form of a white solid.

[0478] 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).

[0479] Step 2-2: Preparation of compound C-7e

[0480] Compound C-7b (22 g, 109.8 mmol) was dissolved in MC (200 mL) at room temperature under a nitrogen atmosphere, and then 3,4-dihydro-2H-pyran (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 C-7e (23.9 g, 76%) in the form of a white solid.

[0481] 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).

[0482] Step 3-2: Preparation of compound C-7f

[0483] Compound C-7e (7.7 g, 27.0 mmol) was dissolved in THF (200 mL) at room temperature under a nitrogen atmosphere, and the mixture was then 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 the mixture was 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 with EA (500 mL), 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 obtain compound C-7f (2.5 g, 27%) in the form of a white solid.

[0484] 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).

[0485] Step 4-2: Preparation of compound C-7g (Method 2)

[0486] Compound C-7f (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, the organic layer was extracted twice, and 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-7g (2.6 g, 89%) in the form of a white solid.

[0487] 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).

[0488] Step 5: Preparation of compound C-7h

[0489] Compound C-7g (2.5 g, 10.9 mmol) was dissolved in DMF (50 mL) at room temperature under a nitrogen atmosphere, and then ethyl chloroacetate (1.40 mL, 13.1 mmol) and potassium carbonate (3.02 g, 21.9 mmol) were added, and the mixture was stirred at 80 ° 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, the organic layer was extracted twice, and a sodium chloride aqueous solution was added to the obtained organic layer, and the organic layer was extracted again. The organic layer after extraction was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue obtained by column chromatography obtained compound C-7h (2.7 gmg, 77%) in the form of a white solid.

[0490] 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).

[0491] Step 6: Preparation of compound C-7i

[0492] Compound C-7h (2.7 g, 8.59 mmol) was dissolved in DMF (50 mL) at room temperature under a nitrogen atmosphere, and potassium carbonate (1.78 g, 12.9 mmol) was then added, 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. The organic layer was extracted twice from the reaction solution with EA (300 mL), and the resulting 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 residue obtained was obtained by column chromatography as a white solid compound C-7i ((2.1 g mg, 82%).

[0493] 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).

[0494] Step 1: Preparation of compound C-7j

[0495] Compound C-7i (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 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 obtain compound C-7j (270 mg, 26.7%).

[0496] 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).

[0497] Step 8: Preparation of Core C-7

[0498] Compound C-7j (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, and EA (100 mL) and distilled water (100 mL) were added at room temperature 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 core C-7 (150 mg, 83%) in the form of a white solid.

[0499] 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).

[0500] Preparation Example 15: Preparation of Core C-8

[0501]

[0502] Step 1: Preparation of compound C-8a

[0503] LAH (2.65 g, 69.77 mmol) was added to THF (350 mL) at 0 ° C under a nitrogen atmosphere, and compound C-7i (8.27 g, 27.91 mmol) 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, and the reaction solution was diluted with EA (300 mL) and filtered with diatomaceous earth. The filtrate was concentrated under reduced pressure at low temperature to reduce the amount of solvent containing THF by half, and then EA (500 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-8a (6.99 g, 98%) in the form of a white solid.

[0504] 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).

[0505] Step 2: Preparation of compound C-8b

[0506] Dess-Martin periodinane (7.02 g, 16.56 mmol) was added to MC (25 mL) at 0 ° C under a nitrogen atmosphere, and then the compound C-8a (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), filtered with diatomaceous earth, and then distilled water (200 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 C-8b (3.31 g, 95%) in the form of a light yellow solid.

[0507] 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).

[0508] Step 3: Preparation of Core C-8

[0509] Compound C-8b (1 g, 3.96 mmol) 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 was subjected to column chromatography to obtain core C-8 (580 mg, 90%) in the form of a white solid.

[0510] 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).

[0511] Preparation Example 16: Preparation of Core C-9

[0512]

[0513] Step 1: Preparation of compound C-9b

[0514] Compound C-9a (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, and then benzyl bromide (400 μL, 3.35 mmol) and potassium carbonate (1.1 g, 7.98 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, EA (100 mL) and distilled water (100 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-9b (750 mg, 95%) in the form of a white solid.

[0515] 1 H-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).

[0516] Step 2: Preparation of compound C-9c

[0517] 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 n-butyl lithium solution (2.5M n-BuLi in hexane, 1.42 mL, 3.56 mmol) was slowly added dropwise. The mixture was stirred at 0 ° C for 40 minutes, compound C-9b (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, the mixture 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. 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-9c (900 mg, 80%) in the form of a light yellow solid.

[0518] 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).

[0519] Step 3: Preparation of compound C-9d

[0520] Compound C-9c (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 for the next reaction without additional purification.

[0521] Step 4: Preparation of compound C-9e

[0522] 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 was subjected to column chromatography to obtain compound C-9e (840 mg, 90%) in the form of a white solid.

[0523] 1H-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] + .

[0524] Step 5: Preparation of compound C-9f

[0525] Compound C-9e (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 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 obtain compound C-9f (310 mg, 72%).

[0526] 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] + .

[0527] Step 6: Preparation of Core C-9

[0528] Compound C-9f (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 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-9 (185 mg, 80%) in the form of a white solid.

[0529] 1H-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).

[0530] Preparation Example 17: Preparation of Core C-10

[0531]

[0532] Step 1: Preparation of compound C-10b

[0533] Compound C-10a (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, and then benzyl chloride (15.78 mL, 137.12 mmol) and potassium carbonate (K2CO3, 22.56 g, 163.24 mmol) were added, and the mixture was stirred at 80°C for 16 hours. After the reaction was completed, EA (200 mL) and distilled water (300 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-10b (31.76 g, 99%) in the form of a light yellow oil.

[0534] 1 H-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).

[0535] Step 2: Preparation of compound C-10c

[0536] Will KO t Bu (15.35 g, 136.82 mmol) was dissolved in THF (100 mL) and 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 added, and the mixture was stirred for 15 minutes. Compound C-10b (31.7 g, 31.7 mmol) was dissolved in THF (50 mL), slowly added to the reaction solution, and the mixture was stirred at room temperature for 19 hours, and then stirred at 80°C for another 3 hours. After the reaction was completed, the reaction solution was cooled to room temperature, ether (200 mL) was added dropwise, the resulting precipitate was filtered, washed with ether (100 mL), and then dried to obtain compound C-10c (41.5 g, 83%) in the form of a light orange solid.

[0537] 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).

[0538] Step 3: Preparation of compound C-10d

[0539] Iron powder (57.1 g, 1.02 mol) was slowly added to acetic acid (200 mL) at room temperature under a nitrogen atmosphere, and compound C-10c (39 g, 102.24 mmol) dissolved in acetic acid (150 mL) was added while stirring, and the mixture was stirred at 80 ° C for 6 hours. After the reaction was completed, the temperature of the reaction solution was reduced to 45 ° C, EA (500 mL) was added dropwise, and the mixture was stirred for another 30 minutes, and the filtrate obtained by filtration through diatomaceous earth was concentrated under reduced pressure. EA (300 mL) was added to the concentrated solution, and the resulting precipitate was filtered again. The filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound C-10d (22.18 g, 73%) in the form of a light yellow solid.

[0540] 1 H-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).

[0541] Step 4: Preparation of Core C-10

[0542] Compound C-10d (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. Then, the mixture was stirred for 3 hours at 60°C to complete the reaction. The temperature of the reaction solution was cooled to room temperature, EA (200 mL) and distilled water (400 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 core C-10 was used for the next step reaction without further purification (6.28 g, 99%).

[0543] 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).

[0544] Preparation Example 18: Preparation of Linker Q-1

[0545]

[0546] The connector P-3 (164 mg, 0.64 mmol) was dissolved in a saturated sodium bicarbonate aqueous solution (3.5 mL) at 0°C under a nitrogen atmosphere, and the mixture was stirred at 0°C for 15 minutes. Compound 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 obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure at a temperature of 15°C or less. The residue thus obtained was subjected to column chromatography to obtain connector Q-1 (85.3 mg, 44%) in the form of a colorless oil.

[0547] 1 H-NMR (400MHz, CDCl3) δ6.70 (s, 2H), 3.74–3.61 (m, 14H), 3.39 (t, J = 5.2Hz, 2H); EI-MS m / z: 321 [M+Na]+ .

[0548] Preparation Example 19: Preparation of Linker Q-2

[0549]

[0550] Step 1: Preparation of compound Q-2b

[0551] Compound Q-2a (bis(2-chloroethyl)amine hydrochloride, CAS No.: 821-48-7, 2 g, 11.20 mmol) was dissolved in distilled water (50 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (1.7 g, 25.77 mmol) was added, and the mixture was stirred at 60 ° C for 48 hours. After the reaction was completed, the mixture was cooled to room temperature, concentrated under reduced pressure, and EA (50 mL) was added to the residue for filtration. The filtrate was concentrated under reduced pressure to obtain compound Q-2b (1.37 g, 78%) in the form of a colorless liquid. The obtained compound C-2b was used for the next step reaction without additional purification process.

[0552] 1 H-NMR (400MHz, CD3OD) δ3.44 (t, J = 6Hz, 4H), 2.78 (t, J = 6Hz, 4H).

[0553] Step 2: Preparation of compound Q-2c

[0554] Compound Q-2b (287 mg, 1.85 mmol) and linker P-2 (487 mg, 1.85 mmol) were dissolved in DMF (10 mL) at 0°C under a nitrogen atmosphere, and then EDCI·HCl (532 mg, 2.77 mmol), HOBt (250 mg, 1.85 mmol), DMAP (22 mg, 0.18 mmol) and DIPEA (1 mL, 5.55 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 solution (50 mL). The organic layer was washed twice by adding a saturated sodium chloride 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 Q-2c (432 mg, 58%) as a colorless oil.

[0555] 1H-NMR (400MHz, CDCl3) δ5.03(brs,1H),4.28(s,2H),3.72–3.68(m,2H),3.67–3.63(m,2H),3.60–3.48(m,10H),3.35–3.30(m,2H),1.45(s,9H); EI-MS m / z:401[M+H] + .

[0556] Step 3: Preparation of compound Q-2b

[0557] Compound Q-2c (432 mg, 1.08 mmol) was dissolved in MC (2 mL) at 0 ° C under a nitrogen atmosphere, and then 4N-hydrochloric acid solution (4M-HCl in dioxane, 4 mL) was added, and the mixture was stirred at 0 ° C for 1 hour and then at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain compound Q-2d (369 mg, 99%) in the form of a light yellow solid. The obtained compound C-2d was used for the next step without additional purification.

[0558] 1 H-NMR(400MHz, CDCl3)δ8.39(brs,2H),4.37(s,2H),3.92–3.87(m,2H),3.77–3.71(m,4H),3. 63–3.56(m,4H),3.54–3.48(m,2H),3.48–3.44(m,2H),3.29–3.21(m,2H); EI-MSm / z:301[M+H] + .

[0559] Step 4: Preparation of linker Q-2

[0560] Compound Q-2d (48 mg, 0.14 mmol) was dissolved in a saturated sodium bicarbonate aqueous solution (1 mL) at 0°C under a nitrogen atmosphere, and the mixture was stirred at 0°C for 15 minutes. Compound Q-1a (TCI, 22 mg, 0.14 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 10°C or lower. The residue thus obtained was subjected to column chromatography to obtain connector Q-2 (17.2 mg, 32%) in the form of a colorless oil.

[0561] 1H-NMR (400MHz, CDCl3) δ6.71(s,2H),4.26(s,2H),3.76–3.72(m,2H),3.67–3.61(m,6H),3.59–3.54(m,6H),3.52–3.48(m,2H).

[0562] Preparation Example 20: Preparation of Linker Q-3

[0563]

[0564] Compound Q-3a (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, and then NHS (49.2 mg, 0.427 mmol) and DCC (88.2 mg, 0.427 mmol) were added, and the mixture was stirred for 15 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, the resulting precipitate was removed, and the filtrate was concentrated under reduced pressure to obtain a joint Q-3 (139 mg, 99%).

[0565] 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] + .

[0566] Preparation Example 21: Preparation of Linker Q-4

[0567]

[0568] Step 1: Preparation of compound Q-4a

[0569] Compound P-4 (4.05 g, 18.47 mmol) was dissolved in ethanol (30 mL) at room temperature under a nitrogen atmosphere, and then 5% palladium on carbon (5% Pd / C, 236 mg, 0.11 mmol) was added, and the mixture was stirred for 16 hours under a hydrogen atmosphere at room temperature. After the reaction was completed, the reaction solution was diluted with EA (250 mL) and filtered with a diatomaceous earth filter to obtain a yellow liquid mixture containing compound Q-4a (3.48 g). The resulting mixture containing Q-4a was used for the next step of the reaction without additional purification.

[0570] 1H-NMR (400MHz, CDCl3) δ3.74–3.69(m,4H),3.69–3.59(m,24H),2.80(t,J=5.2Hz,4H); EI-MS m / z:370[M+H] + .

[0571] Step 2: Preparation of compound Q-4b

[0572] Compound Q-4a (3.48 g, 9.42 mmol) was dissolved in MC (60 mL) at 0 ° C under a nitrogen atmosphere, TEA (4 mL, 28.26 mmol) and di-tert-butyl dicarbonate (Boc anhydride, 3.08 g, 14.13 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 three times with MC (150 mL), distilled water (100 mL) and saturated sodium chloride 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 obtain Q-4b (1.32 g, 32%, 2-step yield) in the form of a colorless oil.

[0573] 1 H-NMR (400MHz, CDCl3) δ3.76–3.69(m,4H),3.69–3.54(m,24H),3.50–3.40(m,4H),2.77(brs,2H),1.45(s,9H); EI-MS m / z:470[M+H] + .

[0574] Step 3: Preparation of compound Q-4c

[0575] Compound Q-4b (595 mg, 1.27 mmol) was dissolved in MC (16 mL) at 0 ° C under a nitrogen atmosphere, and then TEA (1.8 mL, 5.07 mmol) and DMAP (31 mg, 0.25 mmol) were added, and the mixture was stirred at the same temperature for 10 minutes. 4-Methylbenzenesulfonyl chloride (966 mg, 5.07 mmol) was added to the reaction solution, and the mixture was stirred at 0 ° C for 10 minutes and then at room temperature for 3.5 hours. After the reaction was completed, the organic layer was washed three times by adding MC (100 mL) and saturated sodium bicarbonate aqueous solution (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 Q-4c (866 mg, 88%) in the form of a light yellow oil.

[0576] 1H-NMR (400MHz, CDCl3) δ7.79 (d, J = 8Hz, 4H), 7.34 (d, J = 8Hz, 4H), 4.16 (t, J = 4.8Hz, 4H), 3.6 8(t,J=4.8Hz,4H),3.61–3.52(m,20H),3.46–3.38(m,4H),2.44(s,6H),1.44(s,9H); EI-MS m / z:778[M+H] + .

[0577] Step 3: Preparation of compound Q-4d

[0578] Compound Q-4c (866 mg, 1.11 mmol) was dissolved in DMF (10 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (289 mg, 4.46 mmol) was added, and the mixture was stirred at 60 ° C for 16 hours. After the reaction was completed, the organic layer was washed seven times by adding MC (150 mL) and 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 residue was subjected to column chromatography to give Q-4d (519 mg, 90%) in the form of a colorless oil.

[0579] 1 H-NMR (400MHz, CDCl3) δ3.71–3.53 (m, 24H), 3.47–3.41 (m, 4H), 3.39 (t, J = 4.8Hz, 4H), 1.45 (s, 9H).

[0580] Step 5: Preparation of linker Q-4e

[0581] Compound Q-4d (99 mg, 0.19 mmol) was dissolved in MC (1.2 mL) at 0°C under a nitrogen atmosphere, and then a 4M-hydrochloric acid solution (4M-HCl in 1.4-dioxane, 0.7 mL) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the process of adding MC (200 mL), diluting, and concentrating under reduced pressure was repeated 5 times to obtain compound Q-4e (86 mg, 99%) in the form of an orange liquid.

[0582] 1 H-NMR (400MHz, CDCl3) δ9.33(brs,1H),3.93(t,J=4.8Hz,4H),3.73–3.65(m,20H),3.41(t,J=4.8Hz,4H),3.28–3.22(m,4H); EI-MS m / z:420[M+H] + .

[0583] Step 6: Preparation of linker Q-4

[0584] Compound Q-4e (47 mg, 0.103 mmol) and compound Q-3 (47.5 mg, 0.134 mmol) were dissolved in THF (5 mL) at 0 ° C under a nitrogen atmosphere, and then DIPEA (53.9 μL, 0.309 mmol) was added, and the mixture was stirred at 0 ° C for 1.5 hours. Pyridine (24.9 μL, 0.309 mmol) was added to the reaction solution, and the mixture was warmed to room temperature and stirred for 16 hours. After the reaction was completed, the organic layer was extracted three times 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 at 10 ° C or lower to obtain compound Q-4 (67 mg, 99%) in the form of a colorless oil. The obtained compound Q-4 was used for the next step without additional purification process; EI-MS m / z: 659 [M+H] + .

[0585] Example I-1: Preparation of Compound A-1

[0586]

[0587] Step 1: Preparation of compound A-1a

[0588] 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 aqueous sodium hydroxide 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-1a (131.3 mg, 55%) in the form of an ivory solid.

[0589] 1H-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).

[0590] Step 2: Preparation of compound A-1b

[0591] Compound A-1a (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 obtain compound A-1b (101.5 mg, 77%) in the form of a yellow solid.

[0592] 1 H-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).

[0593] Step 3: Preparation of compound A-1c

[0594] Compound A-1b (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 (300 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 obtained residue was subjected to column chromatography to obtain compound A-1c (44.1 mg, 36%).

[0595] 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).

[0596] Step 4: Preparation of Compound A-1

[0597] Compound A-1c (37.2 mg, 0.054 mmol) was dissolved in methanol (1.5 mL) and acetonitrile (1.5 mL) at 0°C under a nitrogen atmosphere, and potassium carbonate (52.4 mg, 0.38 mmol) was then 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 purified by preparative HPLC and lyophilized to obtain compound A-1 (5 mg, 18%) as a white solid.

[0598] 1H-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).

[0599] Example I-2: Preparation of Compound A-2

[0600]

[0601] Step 1: Preparation of compound A-2a

[0602] Core C-2 (160 mg, 0.89 mmol) was dissolved in acetonitrile (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 with 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 obtain compound A-2a (400 mg, 87.7%) in solid form.

[0603] 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).

[0604] Step 2: Preparation of compound A-2b

[0605] Compound A-2a (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-2b (137.2 mg, 75%) in the form of a white solid.

[0606] 1 H-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).

[0607] Step 3: Preparation of compound A-2c

[0608] Compound A-12b (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 the temperature was slowly raised from 0 ° C to room temperature. After the reaction was completed, EA (50 mL) and distilled water (50 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-2c (114.7 mg, 63%) in the form of a white solid.

[0609] 1H-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).

[0610] Step 4: Preparation of compound A-2d

[0611] Compound A-2c (64 mg, 0.094 mmol) was dissolved in DMF (3 mL) at 0 ° C under a nitrogen atmosphere, and 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, it was extracted with EA (50 mL) and 1N-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-2d (49 mg, 68.3%).

[0612] 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] + .

[0613] Step 5: Preparation of compound A-2

[0614] Compound A-2d (49 mg, 0.065 mmol) was dissolved in methanol (2 mL) at 0°C, and then 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 purified by preparative HPLC and freeze-dried to obtain compound A-2 (28.5 mg, 74%) in the form of a white solid.

[0615] 1 H-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] + .

[0616] Example I-3: Preparation of Compound A-3

[0617]

[0618] In the same manner as in Example I-2, compound A-3 (10.4 mg, 65%; EI-MS m / z: 1101 [M+H]) was obtained as a white solid. + ), except that MMAF-OMe was used instead of compound PL-1 in step 4 of Example I-2.

[0619] Example I-4: Preparation of Compound A-4

[0620]

[0621] Using core C-4 as a starting material, compound A-4 (11.7 mg, 64%; EI-MS m / z: 1145 [M+H]) was obtained in the same manner as Example I-2 as a white solid. + ), except that MMAF-OMe was used instead of compound PL-1 in step 4 of Example I-2.

[0622] Example I-5: Preparation of Compound A-5

[0623]

[0624] Using core C-5 as a starting material, compound A-5 (7.7 mg, 60%; EI-MS m / z: 1172 [M+H]) was obtained in the same manner as Example I-2 as a white solid. + ), except that MMAF-OMe was used instead of compound PL-1 in step 4 of Example I-2.

[0625]

[0626] Step 1: Preparation of compound A-6a

[0627] Core C-3 (174 mg, 0.976 mmol) was dissolved in MC (30 mL) at room temperature under a nitrogen atmosphere, and then 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) were added in sequence. 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, 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-6a (250 mg, 50.4%).

[0628] 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).

[0629] Step 2: Preparation of compound A-6b

[0630] Compound A-6a (250 mg, 0.49 mmol) was dissolved in THF (10 mL) at 0 ° C under a nitrogen atmosphere, and then 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 slowly added dropwise to terminate the reaction, and EA (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 obtain compound A-6b (230 mg, 92%).

[0631] 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).

[0632] Step 3: Preparation of compound A-6c

[0633] Compound A-6b (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, and the organic layer was extracted. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain compound A-6c (300 mg, 96.7%) in the form of a white solid; MS m / z: 529 [M+H] + .

[0634] Step 4: Preparation of compound A-6d

[0635] Compound A-6c (300 mg, 0.56 mmol) was dissolved in DMF (15 mL) at room temperature under a nitrogen atmosphere, and then sodium azide (55 mg, 0.84 mmol) was added, and the mixture was stirred at 60 ° C for 2 hours. After the reaction was completed, EA (100 mL) and distilled water (200 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-6d (230 mg, 75.6%).

[0636] 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] + .

[0637] Step 5: Preparation of compound A-6e

[0638] A-6d (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 2 hours. 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-6e (130 mg, 53.7%) in the form of a white solid.

[0639] 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] + .

[0640] Step 6: Preparation of compound A-6f

[0641] Compound A-6e (130 mg, 0.23 mmol) was dissolved in THF (10 mL) at 0 ° C under a nitrogen atmosphere, and then sodium borohydride (22 mg, 0.575 mmol) was added, and the mixture was stirred for 1 hour. 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 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-6f (120 mg, 92.3%).

[0642] 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).

[0643] Step 7: Preparation of compound A-6g

[0644] Compound A-6f (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 obtain compound A-6g (116.3 mg, 75%).

[0645] 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).

[0646] Step 8: Preparation of compound A-6h

[0647] Compound A-6g (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 obtain compound A-6h (17.9 mg, 86%) in the form of a white solid.

[0648] 1 H-NMR (400MHz, CDCl3) δ7.37-7.32(m,1H),7.28–7.22(m,2H),7.19–6.85(m,5H),5.6 1(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] + .

[0649] Step 9: Preparation of compound A-6

[0650] Compound A-6h (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 then 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 purified by preparative HPLC and freeze-dried to obtain compound A-6 (7.5 mg, 53%) in the form of a white solid; EI-MS m / z: 644 [M+H] + .

[0651] Example I-7: Preparation of Compound A-7

[0652]

[0653] Steps 1 and 2: Preparation of Compound A-7b

[0654] In the same manner as Example I-6, compound A-7b (22 mg, 71%; EI-MS m / z: 1156 [M+H]) was obtained as a white solid. + ), except that MMAF-OMe is used instead of compound PL-1 in step 8 of Example I-6, and lithium hydroxide monohydrate is used instead of potassium carbonate in step 9.

[0655] Step 3: Preparation of compound A-7

[0656] Compound A-7b (17.8 mg, 0.0154 mmol) was dissolved in THF (1 mL) at room temperature under a nitrogen atmosphere, and then 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-sodium hydroxide aqueous solution (10 μL) was added to the above reaction solution at room temperature, and the mixture was stirred for 30 minutes, and then 2N-hydrochloric acid aqueous solution was slowly added dropwise to adjust the pH of the reaction solution to 3. The reaction solution was purified by preparative HPLC and then freeze-dried to obtain compound A-7 (11.3 mg, 65%); MS m / z: 1130 [M+H] + .

[0657] Example I-8: Preparation of Compound A-8

[0658]

[0659] Compound A-7b (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 containing a linker P-1 (16.7 mg) dissolved in DMSO (1 mL) was added to the reaction. In addition, distilled water (2 mL), DMSO (0.4 mL), copper 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), purified by preparative HPLC, and then lyophilized to obtain compound A-8 (3.6 mg, 72%); MS m / z: 1357.79 [M+H] + .

[0660] Example I-9: Preparation of Compound A-9

[0661]

[0662] Step 1: Preparation of compound A-9a

[0663] Compound A-4a (114 mg, 0.2 mmol) was dissolved in THF (2 mL) at room temperature under a nitrogen atmosphere, and propargyl bromide (35.8 μL, 0.4 mmol), 1,2-diiodoethane (85 mg, 0.3 mmol) and zinc (Zn powder, 32.9 mg, 0.5 mmol) were added in sequence, and then ultrasonicated for 30 minutes. The mixture was cooled to 0 ° C and then stirred at 0 ° C for 30 minutes. The mixture was warmed to room temperature and stirred for 5 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and filtered through a diatomaceous earth filter. The filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-9a (76.9 mg, 63%) in the form of a white solid.

[0664] 1 H-NMR (400MHz, CDCl3) δ8.12(s,1H),7.45(m,1H),7.01(m,1H),5.62(dd,J=10.4,8Hz,1H),5.48(d,J=3.2Hz,1H),5.16–5.13(m,3H),4.2 7–3.95(m,3H),3.95(s,3H),2.82–2.80(m,2H),2.61(m,1H),2.21(s,3H),2.11(t,J=2.8Hz,1H),2.09(s,3H),2.07(s,3H),2.04(s,3H).

[0665] Step 2: Preparation of compound A-9b

[0666] Compound A-9a (76.9, 0.127mmol) was dissolved in MC (2mL) at 0°C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (51.1mg, 0.253mmol) and DIPEA (66.2μL, 0.38mmol) 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 with EA (100mL) and distilled water (100mL). 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-9b (53.4mg, 54%).

[0667] 1 H-NMR(400MHz, CDCl3)δ8.25(d,J=9.2Hz,2H),8.13(s,1H),7.50(d,J=8.4Hz,1H), 7.36(d,J=9.2Hz,2H),7.02(d,J=8.4Hz,1H),6.05(t,J=6.4Hz,1H),5.62(dd,J=10 .4,8Hz,1H),5.50(d,J=3.2Hz,1H),5.20–5.13(m,2H),4.27–4.09(m,3H),3.95(s, 3H),3.12–2.99(m,2H),2.21(s,3H),2.09(m,4H),2.07(s,3H),2.04(s,3H); EI-MS m / z:794[M+Na] + .

[0668] Step 3: Preparation of compound A-9c

[0669] Compound A-9b (23.8 mg, 0.031 mmol) and compound PL-1 (8.1 mg, 0.037 mmol) were dissolved in DMF (250 μL) at 0 ° C under a nitrogen atmosphere, and then HOBt (6.2 mg, 0.046 mmol), pyridine (250 μL) and DIPEA (13.4 μL, 0.077 mmol) were added in sequence, and the mixture was stirred at room temperature for 5 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 obtain compound A-9c (23.9 mg, 90%) in the form of a white solid; EI-MS m / z: 853 [M + H] + .

[0670] Step 4: Preparation of compound A-9

[0671] Compound A-9c (23.9 mg, 0.028 mmol) was dissolved in methanol (2 mL) and THF (1 mL) at 0°C, and then lithium hydroxide hydrate (11.7 mg, 0.28 mmol) dissolved in distilled water (0.4 mL) was slowly added dropwise, and the mixture was stirred for 4.5 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 purified by preparative HPLC and freeze-dried to obtain compound A-9 (8 mg, 42%) in the form of a white solid; EI-MS m / z: 671 [M+H] + .

[0672] Example I-10: Preparation of Compound A-10

[0673]

[0674] In the same manner as Example I-9, compound A-10 (9.8 mg, 64%; EI-MS m / z: 699 [M+H]) was obtained as a white solid. + ), except that compound A-5a is used instead of the core material in Example I-9.

[0675] Example I-11: Preparation of Compound A-11

[0676]

[0677] Step 1: Preparation of compound A-11a

[0678] Compound A-9b (31.6 mg, 0.041 mmol) was dissolved in DMF (700 μL) at 0 ° C under a nitrogen atmosphere, and MMAF-OMe (monomethyl auristatin F methyl ester, CAS No.: 863971-12-4, 27.8 mg, 0.037 mmol), HOBt (7.5 mg, 0.056 mmol), pyridine (700 μL) and DIPEA (16.2 μL, 0.093 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 obtain compound A-11a (37.2 mg, 72%) in the form of a white solid; EI-MS m / z: 1379 [M+H] + .

[0679] Step 2: Preparation of compound A-11

[0680] Compound A-11a (37.2 mg, 0.027 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C, and then lithium hydroxide hydrate (17 mg, 0.405 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 purified by preparative HPLC and freeze-dried to obtain compound A-11 (20.4 mg, 64%) in the form of a white solid; EI-MS m / z: 1183 [M+H] + .

[0681] Example I-12: Preparation of Compound A-12

[0682]

[0683] Step 1: Preparation of compound A-12a

[0684] Compound A-10b (40 mg, 0.049 mmol) was dissolved in DMF (1 mL) at room temperature under a nitrogen atmosphere, MMAF-OMe (36.7 mg, 0.049 mmol) was added, and the mixture was cooled to 0 ° C. HOBt (10.0 mg, 0.073 mmol), pyridine (1 mL) and DIPEA (21.0 μL, 0.012 mmol) were added in sequence, the mixture was warmed to room temperature and stirred for 16 hours. After the reaction was completed, the reaction solution was diluted with EA (20 mL), and the organic layer was extracted twice with distilled water (20 mL) and 2N-hydrochloric acid aqueous solution (5 mL). 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 A-12a (19 mg, 27%); EI-MS m / z: 1421 [M+H] + .

[0685] Step 2: Preparation of compound A-12

[0686] Compound A-12a (19.7 mg, 0.014 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C, and then lithium hydroxide hydrate (8.7 mg, 0.208 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), and then purified by preparative HPLC and freeze-dried to obtain compound A-12 (11.7 mg, 70%) in the form of a white solid; EI-MS m / z: 1211 [M+H] + .

[0687] Example I-13: Preparation of Compound A-13

[0688]

[0689] Step 1: Preparation of compound A-13b

[0690] Core C-6b (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 (Merck, CAS No.: 30668-32-4, 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 slowly warming 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-13b (1.017 g, 74%) in solid form.

[0691] 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).

[0692] Step 2: Preparation of compound A-13c

[0693] Compound A-13b (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, and the mixture was stirred at -10 ° C 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 obtain compound A-13c (190 mg, 25.8%).

[0694] 1 H-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).

[0695] Steps 3 to 6: Preparation of Compound A-13

[0696] In the same manner as Example I-9, compound A-13 (5.8 mg, 27%; EI-MS m / z: 719 [M+Na]) was obtained as a white solid. + ), except that compound A-13c is used instead of compound A-4a in step 1 of example I-9.

[0697] Example I-14: Preparation of Compound A-14

[0698]

[0699] Using core C-7, compound A-14 (3.5 mg, 15%; EI-MS m / z: 655 [M+Na]) was obtained in a similar manner to Example I-13 as a white solid. + ).

[0700] Example I-15: Preparation of Compound A-15

[0701]

[0702] Compound A-15 (20.4 mg, 46%; EI-MS m / z: 1167 [M+Na]) was obtained in the same manner as in Example I-11 as a white solid. + ), except that compound A-14c is used instead of compound A-9b in step 1 of example I-11.

[0703] Example I-16: Preparation of Compound A-16

[0704]

[0705] In the same manner as Example I-11, compound A-16 (9.3 mg, 61%; EI-MS m / z: 1209 [M+H]) was obtained as a white solid. + ), except that compound A-13e is used instead of compound A-9b in step 1 of example I-11.

[0706] Example I-17: Preparation of Compound A-17

[0707]

[0708] Step 1: Preparation of compound A-17b

[0709] Compound A-17a (120 mg, 0.19 mmol) was dissolved in anhydrous THF (3 mL) at room temperature under a nitrogen atmosphere, and then Zn powder (CAS No.: 7440-66-6, DAEJUNG, 62.5 mg, 0.95 mmol), 1,2-diiodoethane (CAS No.: 624-73-7, Alfa Aesar, 53.9 mg, 0.19 mmol) and propargyl bromide (80% in toluene, 27.1 μL, 0.285 mmol) were added in sequence, and the mixture was ultrasonically reacted in an ultrasonic cleaner (powersonic 410) for 30 minutes. After the reaction was completed, the reaction solution was diluted with EA (50 mL), filtered with diatomaceous earth, and extracted by adding distilled water (50 mL) and 2N-hydrochloric acid aqueous solution (0.7 mL) to the filtered 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 compound A-17b (115 mg, 90%) as a white solid.

[0710] 1H-NMR (400MHz, CDCl3) δ8.38 (d, J=2.4Hz, 1H), 7.29 (m, 1H), 7.04 (dd, J=8, 1.2Hz, 1H ),5.62(dd,J=9.2,8Hz,1H),5.47(d,J=3.6Hz,1H),5.15-5.08(m,3H),4.38(q,J=7. 2Hz,2H),4.29-4.23(m,1H),4.20-4.11(m,2H),2.80-2.71(m,2H),2.63(d,J=2.8Hz ,1H),2.21(s,3H),2.11(m,3H),2.07(m,3H),2.03(s,3H),1.40(t,J=7.2Hz,3H);MS m / z:690[M+H] + .

[0711] Step 2: Preparation of compound A-17c

[0712] Compound A-17b (115 mg, 0.17 mmol) was dissolved in MC (3 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (69.5 mg, 0.34 mmol), pyridine (41.6 μL, 051 mmol) and DIPEA (30 μL, 0.17 mmol) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL). The organic layer was extracted by adding distilled water (100 mL) and 2N-hydrochloric acid aqueous solution (10 mL), dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give A-17c (113 mg, 79%).

[0713] 1 H-NMR (400MHz, CDCl3) δ8.39(s,1H),8.25(d,J=9.2Hz,2H),7.42(d,J=8Hz,1H),7.36(d,J= 9.2Hz,2H),7.07(d,J=8Hz,1H),5.96(t,J=6.4Hz,1H),5.62(dd,J=10.4,8Hz,1H),5.48(d, J=3.2Hz,1H),5.15-5.13(m,2H),4.37(m,2H),4.27-4.12(m,1H),4.19-4.08(m,2H),3.13- 2.95(m,2H),2.21(s,3H),2.10(s,3H),2.07(s,3H),2.03(s,3H),1.41(t,J=7.2Hz,3H); MS m / z:855[M+Na] + .

[0714] Step 3: Preparation of compound A-17d

[0715] Compound A-17c (45 mg, 0.054 mmol) was dissolved in DMF (3 mL) at 0 ° C under a nitrogen atmosphere, and then MMAF-OMe (59.3 mg, 0.081 mmol), HOBt (73 mg, 0.054 mmol), pyridine (300 μL) and DIPEA (19 μL, 0.108 mmol) were added in sequence, and the mixture was stirred for 30 minutes. In addition, the reaction mixture was stirred at room temperature for 20 hours, and then extracted with EA (50 mL) and 1N-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 A-17d (47 mg, 60.4%); EI-MS m / z: 1440 [M+H] + .

[0716] Step 4: Preparation of compound A-17

[0717] Compound A-17d (47 mg, 0.032 mmol) was dissolved in methanol (2 mL) and THF (1 mL) at 0°C, and then lithium hydroxide hydrate (27.4 mg, 0.64 mmol) dissolved in distilled water (0.6 mL) was added dropwise, and the mixture was stirred for 5 hours. After the reaction was completed, 2N-hydrochloric acid aqueous solution (100 μL) was added to terminate the reaction, and the reaction solution was diluted with ACN (1 mL) containing 5% formic acid and distilled water (1 mL), and then purified by preparative HPLC and freeze-dried to obtain compound A-17 (34 mg, 84%) in the form of a white solid; EI-MS m / z: 1230 [M+H] + .

[0718] Example I-18: Preparation of Compound A-18

[0719]

[0720] Compound A-18 (12.3 mg, 59%; EI-MS m / z: 589 [M+H]) was obtained in the same manner as steps 3 to 5 of Example I-2 as a white solid. + ), except that compound A-6b is used instead of compound A-2b in step 3 of example I-2.

[0721] Example I-19: Preparation of Compound A-19

[0722]

[0723] Using core A-13c as a starting material, compound A-19 (8 mg, 58%; EI-MS m / z: 1171 [M+H]) was obtained in the same manner as Example I-2 as a white solid. + ), except that MMAF-OMe was used instead of compound PL-1 in step 4 of Example I-2.

[0724] Example I-20: Preparation of Compound A-20

[0725]

[0726] Using core A-14a as a starting material, compound A-20 (12.1 mg, 68%; EI-MS m / z: 1129 [M+H]) was obtained in the same manner as Example I-2 as a white solid. + ), except that MMAF-OMe was used instead of compound PL-1 in step 4 of Example I-2.

[0727] Example I-21: Preparation of Compound A-21

[0728]

[0729] Using core C-8 as a starting material, compound A-21 (17.2 mg, 74%; EI-MS m / z: 1085 [M+H]) was obtained in the same manner as Example I-2 as a white solid. + ), except that MMAF-OMe was used instead of compound PL-1 in step 4 of Example I-2.

[0730] Example I-22: Preparation of Compound A-22

[0731]

[0732] Steps 1 to 7: Preparation of Compound A-22g

[0733] Compound A-22g (21.3 mg, 61%; EI-MS m / z: 1140 [M+H]) was obtained in the same manner as steps 3 to 8 of Example I-6 as a white solid. + ), except that compound A-21b is used instead of compound A-6b in step 3 of example I-6, and MMAF-OMe is used instead of compound PL-1 in step 8 of example I-6.

[0734] Step 8: Preparation of Compound A-22

[0735] Compound A-22g (15.7 mg, 0.013 mmol) was dissolved in THF (1 mL) at room temperature under a nitrogen atmosphere, and then 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-sodium hydroxide aqueous solution (10 μL) was added to the above reaction solution at room temperature, and the mixture was stirred for another hour, and then 2N-hydrochloric acid aqueous solution was slowly added dropwise to adjust the pH of the reaction solution to 3. The reaction solution was purified by preparative HPLC and then freeze-dried to obtain compound A-22 (10.5 mg, 65%) in the form of a white solid; MS m / z: 1113 [M+H] + .

[0736] Example I-23: Preparation of Compound A-23

[0737]

[0738] Using core C-9 as a starting material, compound A-23 (15.9 mg, 64%; EI-MS m / z: 1192 [M+H]) was obtained in the same manner as Example I-2 as a white solid. + ), except that IPA and chloroform were used as solvent instead of compound THF in step 2 of Example I-2, silica gel was added together with sodium borohydride, and MMAF-OMe was used instead of compound PL-1 in step 4.

[0739] Example I-24: Preparation of Compound A-24

[0740]

[0741] Step 1: Preparation of compound A-24a

[0742] Lithium aluminum hydride (144 mg, 3.8 mmol) was added to THF (15 mL) at 0°C under a nitrogen atmosphere, and compound C-7j (352 mg, 1.08 mmol) dissolved in THF (50 mL) was slowly added, and the mixture was stirred at 0°C for 3 hours. After the reaction was completed, distilled water (100 mL) was added to terminate the reaction, and the reaction solution was diluted with EA (100 mL) and filtered with diatomaceous earth. The obtained solution was concentrated under reduced pressure to remove the organic layer, and then 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-24a (241 mg, 78%) in the form of an ivory solid.

[0743] 1H-NMR (400MHz, CDCl3) δ7.47–7.33(m,5H),7.17(d,J=8Hz,1H),6.84(s,1H),6.70(d,J=8Hz,1H),5 .20(s,2H),4.92(d,J=6Hz,2H),4.78(d,J=6.4Hz,2H),1.87(t,J=6.4Hz,1H),1.79(t,J=6Hz,1H).

[0744] Step 2: Preparation of Compound A-24b

[0745] DMP (Dess-Martin periodinane, 1.3 g, 3.08 mmol) was added to MC (10 mL) at 0 ° C under a nitrogen atmosphere, and then the compound A-24a (350 mg, 1.23 mmol) dissolved in MC (10 mL) and ACN (20 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 (50 mL), filtered with diatomaceous earth, and then distilled water (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 obtain compound A-24b (292 mg, 85%) in the form of an ivory solid.

[0746] 1 H-NMR (400MHz, CDCl3) δ10.40(s,1H),9.92(s,1H),8.01(d,J=8.4Hz,1H),7.75(s,1H),7.48–7.39(m,5H),6.91(d,J=8.4Hz,1H),5.32(s,2H); EI-MS m / z:281[M+H] + .

[0747] Step 3: Preparation of compound A-24c

[0748] Compound A-24b (1.90 mg, 0.68 mmol) was dissolved in MC (15 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 2.03 mL, 2.03 mmol) was slowly added, and the mixture was stirred at -78 ° C for 1.5 hours. After the reaction was completed, distilled water (50 mL) was slowly added dropwise to terminate the reaction, and EA (50 mL) was added to extract the organic layer three times. The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure at a temperature of 15 ° C or lower. The obtained residue was subjected to column chromatography to obtain compound A-24c (56 mg, 43%) in the form of a yellow solid.

[0749] 1 H-NMR (400MHz, DMSO-d6) δ10.09(s,1H),9.84(s,1H),8.07(s,1H),7.95(d,J=8.4Hz,1H),6.88(d,J=8.4Hz,1H).

[0750] Step 4: Preparation of compound A-24d

[0751] Compound A-24c (71.4 mg, 0.37 mmol) was dissolved in MC (13 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (185.3 mg, 0.45 mmol) and benzyltributylammonium chloride (117.1 g, 0.37 mmol) were added. 5N-sodium hydroxide aqueous solution (0.3 mL, 1.5 mmol) was slowly added to the reaction solution, and the mixture was stirred at 0 ° C for 30 minutes and then at room temperature for 8 hours. After the reaction was completed, MC (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. The obtained residue was subjected to column chromatography to obtain compound A-24d (71 mg, 36%) in the form of a white solid.

[0752] 1 H-NMR (400MHz, CDCl3) δ10.46(s,1H),9.37(s,1H),8.03(d,J=8.4Hz,1H),7.63(s,1H),6.99(d,J=8.4Hz,1H),5.62(dd,J=10.4,8Hz,1H), 5.52(m,1H),5.31(d,J=8.0Hz,1H),5.19(dd,J=10.4,3.4Hz,1H),4.17(m,3H),2.21(s,3H),2.08(s,3H),2.07(s,3H),2.04(s,3H); EI-MS m / z:543[M+Na] + .

[0753] Step 5: Preparation of Compound A-24e

[0754] Compound A-24d (71 mg, 0.13 mmol) was dissolved in THF (7 mL) at 0 ° C under a nitrogen atmosphere, and then sodium borohydride (23 mg, 0.61 mmol) was added, and the mixture was stirred at 0 ° C for 2.5 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 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-24e (35.5 mg, 50%) in the form of a white solid.

[0755] 1 H-NMR (400MHz, CDCl3) δ7.21 (d, J = 8Hz, 1H), 6.82 (d, J = 8Hz, 1H), 6.71 (s, 1H), 5.56 (dd, J = 10. 4,8Hz,1H),5.48(d,J=2.8Hz,1H),5.13(dd,J=10.4,3.6Hz,1H),5.10(d,J=7.6Hz,1H),4.94( d,J=6Hz,2H),4.77(d,J=6.4Hz,2H),4.26(dd,J=11.2,6.8Hz,1H),4.18–4.08(m,2H),2.20(s ,3H),2.06(s,3H),2.05(s,3H),2.03(s,3H),2.01(t,J=6.4Hz,1H),1.86(t,J=6Hz,1H); EI-MS m / z:547[M+Na] + .

[0756] Step 6: Preparation of Compound A-24f

[0757] Compound A-24e (35.5 mg, 0.067 mmol) was dissolved in MC (4 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (54.6 mg, 0.271 mmol), pyridine (32.7 μL, 0.406 mmol) and DIPEA (35 μL, 0.203 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), distilled water (50 mL) and 2N-hydrochloric acid aqueous solution (10 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 A-24f (39.7 mg, 69%) in the form of a white viscous gel.

[0758] 1H-NMR(400MHz, CDCl3)δ8.29–8.25(m,4H),7.41–7.37(m,5H),6.97(s,1H),6 .87(d,J=8.4Hz,1H),5.62–5.55(m,3H),5.50(d,J=2.8Hz,1H),5.41(m,2H), 5.17(d,J=7.6Hz,1H),5.15(dd,J=10.4,3.2Hz,1H),4.26(dd,J=11.2,6.8Hz ,1H),4.19–4.09(m,2H),2.20(s,3H),2.07(s,3H),2.05(s,3H),2.03(s,3H).

[0759] Step 7: Preparation of Compound A-24g

[0760] Compound A-24f (12.4 mg, 0.014 mmol) was dissolved in DMF (500 μL) at 0 ° C under a nitrogen atmosphere, and MMAF-OMe (25 mg, 0.033 mmol), HOBt (6 mg, 0.043 mmol), pyridine (500 μL) and DIPEA (12.6 μL, 0.072 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 obtain A-24g (20.7 mg, 70%) in the form of a white solid; EI-MS m / z: 2068 [M+H] + .

[0761] Step 8: Preparation of Compound A-24

[0762] Compound A-24g (20.7 mg, 0.01 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C, and then lithium hydroxide hydrate (6.3 mg, 0.15 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), and then purified by preparative HPLC and freeze-dried to obtain compound A-24 (12 mg, 64%) in the form of a white solid; EI-MS m / z: 1873 [M+H] + .

[0763] Example I-25: Preparation of Compound A-25

[0764]

[0765] Step 1: Preparation of compound A-25a

[0766] Compound A-4b (180 mg, 0.31 mmol) was dissolved in MC (20 mL) at 0 ° C under a nitrogen atmosphere, and then thionyl chloride (48.2 μL, 0.65 mmol) was slowly added, and the mixture was stirred at 0 ° C for 2 hours. After the reaction was completed, the mixture was diluted by adding MC (50 mL), and 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-25a in the form of a white solid. The obtained compound was used for the next step reaction without additional purification process (185 mg, 99%).

[0767] 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] + .

[0768] Steps 2 and 3: Preparation of Compound A-25

[0769] Compound A-25a (30 mg, 0.051 mmol) was dissolved in DMF (3 mL) at 0 ° C under a nitrogen atmosphere, and 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 to the reaction in sequence, and the mixture was stirred at 40 ° C for 16 hours. After the reaction was completed, the generated compound A-25b (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) and 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, and the reaction solution was diluted with ACN (1 mL) and distilled water (1 mL), then purified by preparative HPLC, and then freeze-dried to obtain compound A-25 (1.1 mg, 2%); EI-MS m / z: 1115 [M+H]+ .

[0770] Example I-26: Preparation of Compound A-26

[0771]

[0772] Step 1: Preparation of compound A-26a

[0773] Compound A-25a (135 mg, 0.229 mmol) was dissolved in acetonitrile (5 mL) at room temperature under a nitrogen atmosphere, and then 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 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-26a (140 mg, 97%).

[0774] 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] + .

[0775] Step 2: Preparation of compound A-26b

[0776] 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 then compound A-26a (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 concentrated under reduced pressure to obtain compound A-26b in the form of a white solid, which was used for the next step reaction without additional purification (35 mg, 24%).

[0777] Steps 3 and 4: Preparation of Compound A-26

[0778] Compound A-26b (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 in sequence, 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-26c. The obtained compound A-26c was dissolved by adding methanol (2 mL) and THF (1 mL), 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-hydrochloric acid aqueous 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 purified by preparative HPLC and lyophilized to obtain compound A-26 (0.4 mg, 1%); EI-MS m / z: 825 [M+H] + .

[0779] Example I-27: Preparation of Compound A-35

[0780]

[0781] Step 1: Preparation of compound A-35a

[0782] Compound C-7j (904 mg, 2.79 mmol) was dissolved in THF (40 mL) at room temperature under a nitrogen atmosphere, and zinc (Zn powder, 1.3 g, 19.88 mmol), 1,2-diiodoethane (1.3 g, 4.61 mmol) and propargyl bromide (760 μL, 6.82 mmol) were added in sequence, followed by ultrasonic treatment for 4.5 hours. After the reaction was completed, the reaction solution was diluted with EA (200 mL) and filtered with diatomaceous earth. Distilled water (150 mL) and 2N-hydrochloric acid aqueous solution (50 mL) were added to the filtered residue, 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-35a (770 mg, 76%) in the form of an ivory solid.

[0783] 1H-NMR (400MHz, CDCl3) δ7.67 (s, 1H), 7.49–7.45 (m, 3H), 7.43–7.38 (m, 2H), 7.38–7.33 (m, 1H), 6.76 (d, J = 8Hz, 1H), 5.39 (m, 1H), 5.2 0(s,2H),4.41(q,J=7.2Hz,2H),2.91(m,1H),2.83(m,1H),2.58(d,J=4.8Hz,1H),2.07(t,J=2.8Hz,1H),1.41(t,J=7.2Hz,3H); EI-MS m / z:364[M+H] + ,387[M+Na] + ,347[M-OH] + .

[0784] Step 2: Preparation of Compound A-35b

[0785] Compound A-35a (770mg, 2.11mmol) was dissolved in MC (9mL) and toluene (27mL) at -78°C under a nitrogen atmosphere, and then DIBAL-H (diisobutylaluminum hydride, 1M in toluene, 6.3mL, 6.33mmol) was slowly added, and the mixture was insulated and stirred for 4 hours. After the reaction was completed, methanol (20mL) was added at the same temperature, and the mixture was stirred for 15 minutes, and then distilled water (10mL) and 2N sodium hydroxide aqueous solution (10mL) were added to terminate the reaction. The reaction solution was diluted by adding EA (250mL) and filtered with diatomaceous earth. Saturated potassium sodium tartrate (200mL) was added to the filtered residue, 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 obtained compound A-35b (507mg, 74%) in the form of a yellow solid by column chromatography.

[0786] 1 H-NMR (400MHz, CDCl3) δ7.48–7.43(m,2H),7.42–7.36(m,2H),7.36–7.31(m,1H),7.27–7.23(m,1H),6.81(s,1H),6.69(d,J=8.4Hz,1H), 5.27(m,1H),5.18(s,2H),4.73(d,J=6Hz,2H),2.91–2.79(m,2H),2.68(d,J=4.4Hz,1H),2.22(t,J=6Hz,1H),2.06(t,J=2.8Hz,1H); EI-MS m / z:323[M+H] + ,345[M+Na] + ,305[M-OH]+ .

[0787] Step 3: Preparation of compound A-35c

[0788] Compound A-35b (507 mg, 1.57 mmol) was dissolved in MC (20 mL) at room temperature under a nitrogen atmosphere, and then TBAI (58 mg, 0.16 mmol) and TEMPO (25 mg, 0.16 mmol), sodium bicarbonate (840 mg, 10 mmol) and potassium carbonate (138 mg, 1 mmol) were dissolved in an aqueous solution in distilled water (20 mL), N-chlorosuccinimide (NCS, 231 mg, 1.73 mmol) were added in sequence, and the mixture was stirred for 3 hours. After the reaction was completed, the organic layer was extracted twice with MC (100 mL) and distilled water (100 mL), and washed twice by adding a saturated sodium chloride aqueous solution (100 mL) to the obtained organic layer. 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-35c (401 mg, 80%) in the form of an ivory solid.

[0789] 1 H-NMR (400MHz, CDCl3) δ9.81(s,1H),7.72(s,1H),7.56(d,J=8.4Hz,1H),7.49–7.34(m,5H),6.79(d,J=8.4Hz ,1H),5.39(m,1H),5.23(s,2H),2.93(m,1H),2.81(m,1H),2.57(d,J=4.4Hz,1H),2.07(t,J=2.8Hz,1H); EI-MS m / z:321[M+H] + ,343[M+Na] + ,303[M-OH] + .

[0790] Step 4: Preparation of Core A-35d

[0791] Compound A-35c (502 mg, 1.57 mmol) was dissolved in MC (65 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 4.7 mL, 4.7 mmol) was slowly added, and the mixture was then insulated and stirred for 2.5 hours. After the reaction was completed, distilled water (50 mL) was slowly added dropwise at -50 ° C to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize. MC (100 mL) and distilled water (100 mL) were continued to be added to the mixture, the organic layer was extracted twice, EA (100 mL) was added to the distilled water layer, and the organic layer was extracted once again. 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-35d (131 mg, 36%) in the form of a yellow solid.

[0792] 1 H-NMR (400MHz, DMSO-d6) δ10.52(brs,1H),9.78(s,1H),7.97(s,1H),7.43(d,J=8Hz, 1H), 6.70 (d, J=8Hz, 1H), 5.59 (d, J=4.4Hz, 1H), 5.08 (m, 1H), 2.75–2.59 (m, 3H); EI-MS m / z:231[M+H] + ,253[M+Na] + ,213[M-OH] + .

[0793] Step 5: Preparation of Core A-35e

[0794] Compound A-35d (131 mg, 0.57 mmol) was dissolved in MC (20 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (281 mg, 0.68 mmol) and benzyltributylammonium chloride (178 mg, 0.54 mmol) were added. 5N sodium hydroxide aqueous solution (0.34 mL, 1.71 mmol) was slowly added to the reaction solution, and the mixture was stirred at 0 ° C for 30 minutes and then stirred at room temperature for 4.5 hours. After the reaction was completed, 2N hydrochloric acid aqueous solution was added to neutralize, and then MC (100 mL) and distilled water (100 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-35e (168 mg, 52%) in the form of a yellow gum.

[0795] 1H-NMR (400MHz, CDCl3) δ9.84 (s, 1H), 7.63–7.56 (m, 2H), 6.90 (m, 1H), 5.59 (dd, J = 10.4, 8Hz, 1H), 5.50 (m, 1H), 5.40 (m, 1 EI-MS m / z:583[M+Na] + .

[0796] Step 6: Preparation of Compound A-35f

[0797] Compound A-35e (168 mg, 0.3 mmol) was dissolved in THF (5 mL) at 0 ° C under a nitrogen atmosphere, and then sodium borohydride (28 mg, 0.75 mmol) was added, and the mixture was stirred for 2.5 hours. After the reaction was completed, distilled water (50 mL) was added at the same temperature 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-35f (119 mg, 70%) in the form of a white solid.

[0798] 1 H-NMR (400MHz, CDCl3) δ7.31(m,1H),6.83(m,1H),6.71(s,1H),5.56(dd,J=10.4,8Hz,1H),5.48(d,J=3.2Hz,1H),5.31(m,1H),5.17–5.09 EI-MS m / z:585[M+Na] + .

[0799] Step 7: Preparation of Compound A-35g

[0800] Compound A-35f (50 mg, 0.089 mmol) was dissolved in MC (2 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (72 mg, 0.356 mmol) and pyridine (43 μL, 0.533 mmol), DIPEA (46.4 μL, 0.267 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 with EA (50 mL) and 2N-hydrochloric acid aqueous solution (50 mL). The organic layer after extraction 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-35g (27 mg, 34%) in solid form.

[0801] 1 H-NMR (400MHz, CDCl3) δ8.33–8.23(m,4H),8.12–8.08(m,1H),7.45–7.34(m,4H),7.01–6.95(m,2H),6.35–6.26(m,1H),5.61–5.53(m,1H EI-MS m / z:915[M+Na] + .

[0802] Step 8: Preparation of Compound A-35h

[0803] Compound A-35g (27 mg, 0.03 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAF-OMe (CAS No.: 863971-12-4, 34 mg, 0.045 mmol), HOBt (12 mg, 0.091 mmol), DIPEA (26.3 μL, 0.151 mmol) and pyridine (1 mL) 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 purified by preparative HPLC and freeze-dried to obtain compound A-35h (16.8 mg, 26%) in the form of a solid; EI-MS m / z: 2107 [M+H] + , 10541 / 2[M+H] + .

[0804] Step 9: Preparation of Compound A-35

[0805] Compound A-35h (16.8 mg, 0.008 mmol) was dissolved in methanol (1 mL) and THF (0.5 mL) at 0°C, and then lithium hydroxide hydrate (5 mg, 0.119 mmol) dissolved in distilled water (0.2 mL) was slowly added dropwise, and the mixture was stirred at 0°C for 1 hour and stirred at room temperature for another 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), and then purified by preparative HPLC and freeze-dried to obtain compound A-35 (12.5 mg, 82%) in the form of a white solid; EI-MS m / z: 1911 [M+H] + ,956 1 / 2[M+H] + .

[0806] Example I-28: Preparation of Compound A-36

[0807]

[0808] Step 1: Preparation of compound A-36a

[0809] Compound C-9e (5.42 mg, 15.09 mmol) was dissolved in MC (60 mL) and toluene (180 mL) at -78 ° C under a nitrogen atmosphere, and then DIBAL-H (diisobutylaluminum hydride, 1 M in toluene, 37.7 mL, 37.7 mmol) was slowly added, and the mixture was insulated and stirred for 2.5 hours. After the reaction was completed, methanol (50 mL) was added and stirred for 15 minutes at the same temperature, and then distilled water (50 mL) and 2N sodium hydroxide aqueous solution (50 mL) were added at 0 ° C to terminate the reaction. The reaction solution was diluted with EA (800 mL) and filtered with diatomaceous earth. Saturated potassium sodium tartrate (300 mL) was added to the filtered residue, 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-36a (4.52 mg, 94%) in the form of an ivory solid.

[0810] 1 H-NMR(400MHz, CDCl3)δ7.65(s,1H),7.50–7.44(m,3H),7.44–7.37(m,2H),7.37–7.32(m,1H),7.18 (t,J=8Hz,1H),6.82(d,J=8Hz,1H),5.18(s,2H),4.92(d,J=5.2Hz,2H),1.96(t,J=5.2Hz,1H);EI-MS m / z:317[M+H] + ,299[M-OH] + .

[0811] Step 2: Preparation of Compound A-36b

[0812] Compound A-36a (4.52 g, 14.25 mmol) was dissolved in MC (92 mL) at room temperature under a nitrogen atmosphere, and then TBAI (526 mg, 1.42 mmol) and TEMPO (223 mg, 1.42 mmol), sodium bicarbonate (3.86 mg, 46 mmol) and potassium carbonate (636 mg, 4.6 mmol) were dissolved in distilled water (92 mL) in sequence. An aqueous solution, N-chlorosuccinimide (NCS, 2.09 g, 15.67 mmol) was added, and the mixture was stirred for 3 hours. After the reaction was completed, the organic layer was extracted twice with MC (500 mL) and distilled water (500 mL), and washed twice by adding a saturated sodium chloride aqueous solution (400 mL) to the obtained organic layer. The washed organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was obtained by column chromatography to obtain compound A-36b (4.13 g, 92%) in the form of a light orange solid.

[0813] 1 H-NMR (400MHz, CDCl3) δ9.94 (s, 1H), 8.53 (s, 1H), 7.53–7.46 (m, 3H), 7.46–7.40 (m, 2H), 7.40–7.34 (m, 2H), 6.87 (d, J = 8Hz, 1H), 5.23 (s, 2H); EI-MS m / z:315[M+H] + .

[0814] Step 3: Preparation of compound A-36c

[0815] Compound A-36b (4.13 g, 13.12 mmol) was dissolved in MC (65 mL) at room temperature under nitrogen atmosphere, (carboethoxymethylene)triphenylphosphorane (combi-blocks, CAS No.: 1099-45-2, 9.14 g, 26.23 mmol) was added, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-36c (4.81 g, 95%) in the form of a pink solid.

[0816] 1H-NMR (400MHz, CDCl3) δ7.93 (s, 1H), 7.85 (d, J = 15.2Hz, 1H), 7.49–7.44 (m, 2H), 7.44–7.38 (m, 3H), 7.38–7.33 (m, 1H), 7.27 EI-MS m / z:385[M+H] + .

[0817] Step 4: Preparation of compound A-36d

[0818] Compound A-36c (4.81 g, 12.48 mmol) was dissolved in EA (140 mL) and methanol (420 mL) at room temperature under a nitrogen atmosphere, and then 5% palladium on carbon (5% Pd / C, 5.1 g) was added, and the mixture was stirred for 2 days under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered through diatomaceous earth and concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-36d (3.87 g, 80%) in the form of a light yellow oil.

[0819] 1 H-NMR (400MHz, CDCl3) δ7.51–7.44(m,3H),7.44–7.37(m,3H),7.37–7.32(m,1H),7.14(t,J=8Hz,1H),6.80(d,J=8Hz ,1H),5.18(s,2H),4.16(q,J=7.2Hz,2H),3.27(t,J=7.6Hz,2H),2.72(t,J=7.6Hz,2H),1.25(t,J=7.2Hz,3H); EI-MS m / z:387[M+H] + .

[0820] Step 5: Preparation of Compound A-36e

[0821] Compound A-36d (3.87 g, 9.99 mmol) was dissolved in MC (285 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (2.74 mL, 29.97 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 29.9 mL, 29.97 mmol) were slowly added in sequence, and the mixture was stirred for 2 hours. After the reaction was completed, distilled water (700 mL) was slowly added dropwise to terminate the reaction, and MC (600 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-36e (2.64 g, 64%).

[0822] 1 H-NMR (400MHz, CDCl3) δ10.04(s,1H),7.72(d,J=8.4Hz,1H),7.58(s,1H),7.51–7.34(m,5H),6.98(d,J=8.4Hz,1 EI-MS m / z:415[M+H] + .

[0823] Step 6: Preparation of Compound A-36f

[0824] Compound A-36e (2.64 g, 6.35 mmol) was dissolved in MC (250 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 19 mL, 19.07 mmol) was slowly added, and the mixture was then insulated and stirred for 2.5 hours. After the reaction was completed, distilled water (150 mL) was slowly added dropwise at -50 ° C to terminate the reaction, and a 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize the reaction solution. MC (250 mL) and distilled water (150 mL) were continued to be 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-36f (1.15 g, 55%) in the form of a light yellow solid.

[0825] 1H-NMR (400MHz, CDCl3) δ10.00 (s, 1H), 7.62 (d, J = 8Hz, 1H), 7.51 (s, 1H), 6.96 (s, 1H), 6.87 (d, J = 8Hz, 1H), 4.19 (q, J = 7.2Hz, 2H), 3.33 (t, J = 7.2Hz, 2H), 2.79 (t, J = 7.2Hz, 2H), 1.27 (t, J = 7.2Hz, 3H); EI-MS m / z:325[M+H] + .

[0826] Step 7: Preparation of Compound A-36g

[0827] Compound A-36f (481 mg, 1.48 mmol) was dissolved in MC (50 mL) at 0°C under a nitrogen atmosphere, and then acetyl bromide-α-D-glucuronic acid methyl ester (TCI, CAS No.: 21085-72-3, 705 mg, 1.77 mmol), benzyltributylammonium chloride (461 mg, 1.48 mmol) and 5N-sodium hydroxide aqueous solution (0.89 mL, 4.43 mmol) were slowly added, and the mixture was stirred at 0°C for 30 minutes, and stirred at room temperature for another 16 hours. After the reaction was completed, 2N hydrochloric acid aqueous solution was added to neutralize the reaction solution, and MC (200 mL) and distilled water (200 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-36g (238 mg, 25%) in the form of an ivory solid.

[0828] 1 H-NMR (400MHz, CDCl3) δ10.10(s,1H),7.74(d,J=8Hz,1H),7.39(s,1H),7.09(d,J=8Hz,1H),5.48–5.37(m,4H),4.32–4.27(m,1H),4.18(q,J =7.2Hz,2H),3.71(s,3H),3.31(t,J=7.6Hz,2H),2.74(t,J=7.6Hz,2H),2.09(s,3H),2.07(s,3H),2.06(s,3H),1.27(t,J=7.2Hz,3H); EI-MS m / z:663[M+Na] + .

[0829] Step 8: Preparation of Compound A-36h

[0830] Compound A-36g (67.6 mg, 0.105 mmol) was dissolved in THF (3 mL) at room temperature under a nitrogen atmosphere, and zinc (Zn powder, 51 mg, 0.79 mmol), 1,2-diiodoethane (59.6 mg, 0.21 mmol) and propargyl bromide (37.4 μL, 0.42 mmol) were added in sequence, and then ultrasonicated for 2.5 hours. After the reaction was completed, the reaction solution was diluted with EA (50 mL) and filtered with diatomaceous earth. Distilled water (30 mL) and saturated sodium chloride aqueous solution (20 mL) were added to the filtered residue, 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-36h (61.8 mg, 86%) in the form of a white gum.

[0831] 1 H-NMR (400MHz, CDCl3) δ7.34(s,1H),7.20–7.15(m,1H),6.93(d,J=8Hz,1H),5.43–5.33(m,3H),5.22–5.18(m,1H),5.01(m,1H),4.22–4.09(m, 3H),3.73(s,3H),3.25(t,J=7.6Hz,2H),2.82–2.68(m,4H),2.52(m,1H),2.11(t,J=2.8Hz,1H),2.10–2.02(m,9H),1.27(t,J=7.2Hz,3H); EI-MS m / z:681[M+H] + ,704[M+Na] + ,663[M-OH] + .

[0832] Step 9: Preparation of Compound A-36i

[0833] Compound A-36h (65.1 mg, 0.095 mmol) was dissolved in MC (3 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (57.7 mg, 0.286 mmol), DIPEA (75 μL, 0.43 mmol), pyridine (23.1 μL, 0.286 mmol), DMAP (2.3 mg, 0.019 mmol) and DMF (20 μL) were added in sequence, and the mixture was stirred for 24 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 (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 obtain compound A-36i (62.6 mg, 77%) in the form of a white solid.

[0834] 1 H-NMR (400MHz, CDCl3) δ8.25 (d, J = 9.2Hz, 2H), 7.39–7.33 (m, 3H), 7.27 (m, 1H ),6.95(d,J=8.4Hz,1H),5.91(m,1H),5.42–5.34(m,3H),5.26–5.21(m,1H), 4.25–4.05(m,3H),3.73(s,3H),3.26(t,J=7.2Hz,2H),3.04(m,1H),2.95(m, 1H),2.72(t,J=7.2Hz,2H),2.09–2.03(m,10H),1.27(t,J=7.2Hz,3H); EI-MS m / z:869[M+Na] + .

[0835] Step 10: Preparation of Compound A-36j

[0836] Compound A-36i (43.5 mg, 0.051 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAF (monomethyl auristatin E, CAS No.: 474645-27-7, 36.9 mg, 0.051 mmol), HOBt (10.4 mg, 0.077 mmol), DIPEA (22.4 μL, 0.128 mmol) and pyridine (1 mL) 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 obtain A-36j (48.5 mg, 66%) in the form of a white gum; EI-MS m / z: 1426 [M+H] + .

[0837] Step 11: Preparation of Compound A-36

[0838] Compound A-36j (48.5 mg, 0.034 mmol) was dissolved in THF (0.34 mL), methanol (0.85 mL) and distilled water (0.22 mL) at -20 ° C, and then lithium hydroxide hydrate (10.7 mg, 0.255 mmol) dissolved in distilled water (0.63 mL) was slowly added dropwise, and the mixture was stirred for 4.5 hours while the temperature was slowly raised from -20 ° C to -5 ° C. The reaction solution was stirred for another 3 hours at 0 ° C. 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 (2 mL) and distilled water (2 mL), and then purified by preparative HPLC and freeze-dried to obtain compound A-36 (29.8 mg, 70%) in the form of a white solid; EI-MS m / z: 1258 [M+H] + .

[0839] Example I-29: Preparation of Compound A-37

[0840]

[0841] Step 1: Preparation of compound A-37a

[0842] Compound A-36i (32.5 mg, 0.038 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAF-OMe (28.6 mg, 0.038 mmol), HOBt (7.8 mg, 0.057 mmol), DIPEA (16.7 μL, 0.096 mmol) and pyridine (1 mL) 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 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 obtain compound A-37a (41.3 mg, 74%) in solid form; EI-MS m / z: 1454 [M+H] + .

[0843] Step 2: Preparation of compound A-37

[0844] Compound A-37a (41.3 mg, 0.028 mmol) was dissolved in THF (0.35 mL), methanol (0.71 mL) and distilled water (0.2 mL) at -20 ° C, and then lithium hydroxide hydrate (8.9 mg, 0.213 mmol) dissolved in distilled water (0.51 mL) was slowly added dropwise, and the mixture was stirred for 2.5 hours while the temperature was slowly raised from -20 ° C to -5 ° C. The reaction solution was stirred for another 3 hours at 0 ° C. 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 (2 mL) and distilled water (2 mL), and then purified by preparative HPLC and freeze-dried to obtain compound A-37 (24.9 mg, 69%) in the form of a white solid; EI-MS m / z: 1272 [M+H] + .

[0845] Example I-30: Preparation of Compound A-38

[0846]

[0847] Step 1: Preparation of compound A-38a

[0848] Compound A-17c (70 mg, 0.084 mmol) was dissolved in DMF (3 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (112.4 mg, 0.10 mmol), HOBt (11.3 mg, 0.084 mmol), pyridine (300 μL) and DIPEA (29.3 μL, 0.168 mmol) were added in sequence, and the mixture was stirred for 30 minutes. The reaction mixture was stirred at room temperature for another 15 hours, and then extracted with EA (50 mL) and 1N-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 A-38a (87 mg, 73.3%); EI-MS m / z: 1412 [M+H] + ,1434[M+Na] + .

[0849] Step 2: Preparation of compound A-38

[0850] Compound A-38a (87 mg, 0.061 mmol) was dissolved in methanol (3 mL) and THF (1.5 mL) at 0 ° C, and then lithium hydroxide hydrate (51.7 mg, 1.22 mmol) dissolved in distilled water (0.6 mL) was added dropwise, and the mixture was stirred for 30 minutes. The reaction mixture was stirred for another 5 hours at room temperature, and then 2N-hydrochloric acid aqueous solution was slowly added to adjust the pH of the reaction solution to 3. The reaction solution was diluted with ACN (5 mL) containing 0.1% formic acid and distilled water (5 mL), and then purified by preparative HPLC and freeze-dried to obtain compound A-38 (59.1 mg, 79%) in the form of a white solid; EI-MS m / z: 1216 [M+H] + .

[0851] Example I-31: Preparation of Compound A-39

[0852]

[0853] Step 1: Preparation of compound A-39b

[0854] THF (12.0 mL) was added dropwise to sodium hydride (NaH, 60% dispersion in mineral oil, 724.0 mg, 18.1 mmol) at 0°C under a nitrogen atmosphere, and compound A-39a (Alfa Aesar, CAS No.: 24677-78-9, 1.0 g, 7.24 mmol) dissolved in THF (8.0 mL) was slowly added to the reaction solution, and the mixture was stirred at room temperature for 1 hour. Benzyl bromide (0.86 mL, 7.24 mmol) was slowly added to the reaction solution, and the mixture was stirred for another 16 hours. After the reaction was completed, distilled water (50 mL), 2N-hydrochloric acid aqueous solution (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. Ethanol (30.0 mL) was added to the residue, and the obtained solid was then filtered to obtain compound A-39b (680.0 mg, 41%) in the form of a yellow solid.

[0855] 1 H-NMR(400MHz, CDCl3)δ11.07(s,1H),9.92(s,1H),7.45–7.31(m,5H),7.19(dd,J =7.6Hz, J=1.2Hz, 1H), 7.13 (d, J = 7.6Hz, 1H), 6.89 (t, J = 7.6Hz, 1H), 5.19 (s, 3H).

[0856] Step 2: Preparation of compound A-39c

[0857] Compound A-39b (680.0 mg, 2.97 mmol) was dissolved in DMF (5.0 mL) under a nitrogen atmosphere, and ethyl bromoacetate (0.38 mL, 3.57 mmol) and potassium carbonate (1.03 g, 7.44 mmol) were added in sequence, and the mixture was stirred at 80 ° C for 2 hours. After the reaction was completed, distilled water (50 mL) and EA (50 mL) were added to extract the organic layer. The obtained organic layer was washed by adding a saturated sodium chloride aqueous solution (50 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was obtained by column chromatography as a white solid compound A-39c (870.0 mg, 93%).

[0858] 1 H-NMR (400MHz, CDCl3) δ10.63(s,1H),7.47(d,J=8.0Hz,1H),7.44–7.34(m,5H),7.19(d,J=8.0Hz, 1H),7.11(t,J=8.0Hz,1H),5.13(s,2H),4.85(s,2H)4.15(q,J=7.2Hz,2H),1.22(t,J=7.2Hz,3H).

[0859] Step 3: Preparation of compound A-39d

[0860] Compound A-39c (870.0 mg, 2.76 mmol) was dissolved in DMF (10.0 mL) under a nitrogen atmosphere, potassium carbonate (1.03 g, 7.44 mmol) was then added, and the mixture was stirred at 100 ° C for 3 hours. After the reaction was completed, distilled water (100 mL) and EA (100 mL) were added to extract the organic layer. The obtained organic layer was washed by adding a saturated aqueous sodium chloride solution (100 mL). The 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-39d (640.0 mg, 78%) in the form of a white solid.

[0861] 1 H-NMR (400MHz, CDCl3) δ7.51–7.49(m,3H),7.40–7.30(m,3H),7.25–7.23(m,1H),7.15(t,J=8.0 Hz, 1H), 6.94 (d, J = 8.0Hz, 1H), 5.33 (s, 2H), 4.44 (q, J = 7.2Hz, 2H), 1.42 (t, J = 7.2Hz, 3H); EI-MS m / z:297[M+H] + .

[0862] Step 4: Preparation of Compound A-39e

[0863] Compound A-39d (640.0 mg, 2.16 mmol) was dissolved in MC (20.0 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (0.57 mL, 6.48 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 6.48 mL, 6.48 mmol) were slowly added in sequence, and the mixture was stirred for 2 hours. 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-39e (420.0 mg, 60%) in the form of a white solid.

[0864] 1 H-NMR (400MHz, CDCl3) δ10.03(s,1H),8.22(s,1H),7.69(d,J=8.0Hz,1H),7.50–7.48(m,2H),7.42– 7.34(m,3H),7.04(d,J=8.0Hz,1H),5.42(s,2H),4.45(q,J=7.2Hz,2H),1.43(t,J=7.2Hz,3H); EI-MS m / z:325[M+H] + .

[0865] Step 5: Preparation of Compound A-39f

[0866] Compound A-39e (420.0 mg, 1.29 mmol) was dissolved in MC (13.0 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 3.88 ml, 3.88 mmol) was slowly added, and the mixture was then insulated and stirred for 2 hours. After the reaction was completed, distilled water (50.0 mL) was slowly added dropwise at -50 ° C to terminate the reaction, and EA (100 mL) and distilled water (100 mL) were added at room temperature 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-39f (145.0 mg, 47%) in the form of an ivory solid.

[0867] 1 H-NMR (400MHz, CDCl3) δ10.05 (s, 1H), 8.23 ​​(s, 1H), 7.73 (d, J = 8.0Hz, 1H), 7.1 0(d,J=8.0Hz,1H),6.51(s,1H),4.46(q,J=7.2Hz,2H),1.44(t,J=7.2Hz,3H).

[0868] Step 6: Preparation of Compound A-39g

[0869] Compound A-39f (140.0 mg, 0.59 mmol) was dissolved in ACN (20.0 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (Merck, 295.0 mg, 0.71 mmol) and silver (I) oxide (346.0 mg, 1.49 mmol) were added in sequence, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL), filtered with diatomaceous earth, and washed by adding saturated sodium chloride aqueous solution (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 obtained by column chromatography as a white solid compound A-39g (303.0 mg, 89%).

[0870] 1 H-NMR (400MHz, CDCl3) δ10.10(s,1H),8.23(s,1H),7.73(d,J=8.0Hz,1H),7.27(d,J=8.0Hz,1H),5.60–5.59(m,2H),5.50(d,J=3.2Hz,1H),5 .21–5.17(m,1H),4.45(q,J=7.2Hz,2H),4.23–4.17(m,3H),2.19(s,3H),2.14(s,3H),2.03(s,3H),1.98(s,3H),1.43(t,J=7.2Hz,3H); EI-MS m / z:586[M+Na] + .

[0871] Step 7: Preparation of Compound A-39h

[0872] Compound A-39g (100.0 mg, 0.2 mmol) was dissolved in THF (2.0 mL) at room temperature under a nitrogen atmosphere, and propargyl bromide (23.6 μL, 0.265 mmol), 1,2-diiodoethane (57.8 mg, 0.177 mmol) and zinc (Zn powder, 57.8 mg, 0.885 mmol) were added in sequence, and then ultrasonicated for 2 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and filtered with diatomaceous earth. The filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-39h (85.0 mg, 79%) as a white solid.

[0873] 1H-NMR (400MHz, CDCl3) δ7.75(d,J=4.0Hz,1H),7.25(d,J=4.0Hz,1H),7.20(s,1H),5.57(dd,J=10.4Hz,J =8.0Hz,1H),5.47(d,J=3.2Hz,1H),5.27(dd,J=8.0Hz,J=2.0Hz,1H),5.15(dd,J=10.4Hz,J=3.2Hz,2H), 4.43(q,J=7.2Hz,2H),4.21–4.18(m,1H)4.11–4.06(m,1H),2.75–2.71(m,2H),2.51(d,J=3.2Hz,1H),2. 19(s,6H),2.10(t,J=3.2Hz,1H),2.02(s,3H),2.01(s,3H),1.42(t,J=7.2Hz,3H),1.25(t,J=7.2Hz,1H).

[0874] Step 8: Preparation of Compound A-39i

[0875] Compound A-39h (85.0 mg, 0.14 mmol) was dissolved in MC (3.0 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (42.5 mg, 0.21 mmol) and DIPEA (25.0 μL, 0.14 mmol) and pyridine (34.0 μL, 0.42 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, EA (50 mL) and distilled water (50 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-39i (56.0 mg, 51%) in the form of a white solid.

[0876] 1H-NMR (400MHz, CDCl3) δ8.25(d,J=9.2Hz,2H),7.72(d,J=0.8Hz,1H),7.34(d,J=9.2Hz,2H),7.31–7.21(m,2 H),6.07(t,J=7.2Hz,1H),5.59(dd,J=10.4Hz,J=8.0Hz,1H),5.48(d,J=3.2Hz,1H),5.33(dd,J=8.0Hz,J=2.0 Hz,1H),5.15(dd,J=10.4Hz,J=3.2Hz,1H),4.44(t,J=7.2Hz,2H),4.22–4.18(m,2H),4.15–4.09(m,2H),3.08 –3.02(m,1H),2.93–2.87(m,1H),2.19(s,3H),2.17(s,3H),2.03(s,3H),2.01(s,3H),1.43(t,J=7.2Hz,3H).

[0877] Step 9: Preparation of Compound A-39j

[0878] Compound A-39i (56.0 mg, 0.072 mmol) was dissolved in DMF (2.0 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (52.2 mg, 0.072 mmol), HOBt (14.7 mg, 0.109 mmol), DIPEA (31.7 μL, 0.181 mmol) and pyridine (1.0 mL) were added in sequence, and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, 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 obtain compound A-39j (56.0 mg, 57%) in the form of a white solid; EI-MSm / z: 1349 [M+H] + .

[0879] Step 10: Preparation of Compound A-39

[0880] Compound A-39j (29.0 mg, 0.021 mmol) was dissolved in methanol (1.0 mL) at 0°C under a nitrogen atmosphere, and then lithium hydroxide (6.8 mg, 0.161 mmol) dissolved in distilled water (1.0 mL) 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 purified by preparative HPLC and freeze-dried to obtain compound A-39 (18.0 mg, 72%) in the form of a white solid; EI-MS m / z: 1153 [M+H] + .

[0881] Example I-32: Preparation of Compound A-40

[0882]

[0883] Step 1: Preparation of compound A-40a

[0884] Core C-5 (500 mg, 1.79 mmol) was dissolved in MC (60 mL) at 0°C under a nitrogen atmosphere, and then acetyl bromide-α-D-glucuronic acid methyl ester (TCI, 856 mg, 2.15 mmol), benzyltributylammonium chloride (560 mg, 1.79 mmol) and 5N-sodium hydroxide aqueous solution (1.08 mL, 5.39 mmol) were slowly 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, 2N hydrochloric acid aqueous solution was added to neutralize the reaction solution, and MC (100 mL) and distilled water (100 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-40a (88.7 mg, 8%) in the form of an ivory solid.

[0885] 1 H-NMR (400MHz, CDCl3) δ10.08(s,1H),7.76(d,J=8Hz,1H),7.16(s,1H),7.05(d,J=8Hz,1H),5.46–5.37(m,4H),4.32–4.28(m,1H),4.16(q,J =7.2Hz,2H),3.71(s,3H),3.27(t,J=7.6Hz,2H),2.76(t,J=7.6Hz,2H),2.09(s,3H),2.07(s,3H),2.06(s,3H),1.27(t,J=7.2Hz,3H); EI-MS m / z:594[M+H] + .

[0886] Step 2: Preparation of Compound A-40b

[0887] Compound A-40a (110 mg, 0.185 mmol) was dissolved in THF (3 mL) at room temperature under a nitrogen atmosphere, and zinc (Zn powder, 60.5 mg, 0.925 mmol), 1,2-diiodoethane (104.2 mg, 0.37 mmol) and propargyl bromide (46.3 μL, 0.416 mmol) were added in sequence, and then ultrasonicated for 3.5 hours. After the reaction was completed, the reaction solution was diluted with EA (50 mL) and filtered with diatomaceous earth. The organic layer was extracted by adding distilled water (30 mL) and saturated sodium chloride aqueous solution (20 mL). The obtained organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was obtained by column chromatography to give compound A-40b (93.1 mg, 79%) in the form of a white solid.

[0888] 1 H-NMR (400MHz, CDCl3) δ7.29(m,1H),7.12(s,1H),6.91(d,J=8.4Hz,1H),5.43–5.33(m,3H),5.24–5.19(m,1H),5.10(m,1H),4.24–4.11(m,3H),3. 73(s,3H),3.22(t,J=7.6Hz,2H),2.81–2.77(m,2H),2.73(t,J=7.6Hz,2H ),2.52(t,J=3.2Hz,1H),2.12–2.02(m,10H),1.27(t,J=7.2Hz,3H); EI-MS m / z:634[M+H] + ,656[M+Na] + ,616[M-OH] + .

[0889] Step 3: Preparation of Compound A-40c

[0890] Compound A-40b (93.1 mg, 0.147 mmol) was dissolved in MC (3 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (59.1 mg, 0.293 mmol) and DIPEA (38.3 μL, 0.22 mmol) and pyridine (35.4 μL, 0.44 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 30 minutes. After the reaction was completed, 2N-hydrochloric acid aqueous solution (10 mL) was added at 0 ° C to acidify the reaction solution, and then the organic layer was extracted once with MC (50 mL) and 2N-hydrochloric acid aqueous solution (40 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 compound A-40c (84.9 mg, 72%) in the form of a white solid.

[0891] 1 H-NMR (400MHz, CDCl3) δ8.25(d,J=9.2Hz,2H),7.36(d,J=9.2Hz,2H),7.32(d,J=8.4Hz,1H ),7.14(s,1H),6.93(d,J=8.4Hz,1H),6.01(t,J=6.4Hz,1H),5.44–5.34(m,3H),5.28–5.2 3(m,1H),4.26–4.09(m,3H),3.73(s,1.5H),3.72(s,1.5H),3.23(t,J=7.6Hz,2H),3.05(m ,1H),2.98(m,1H),2.74(t,J=7.6Hz,2H),2.09–2.03(m,10H),1.26(t,J=7.2Hz,3H); EI-MS m / z:821[M+Na] + .

[0892] Step 4: Preparation of Compound A-40d

[0893] Compound A-40c (47.5 mg, 0.059 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAF-OMe (44.3 mg, 0.059 mmol), HOBt (12 mg, 0.089 mmol), DIPEA (25.8 μL, 0.148 mmol) and pyridine (1 mL) 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), and washed by adding saturated sodium chloride 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 A-40d (69.2 mg, 83%) in the form of a white gum; EI-MS m / z: 1407 [M+H]+ .

[0894] Step 5: Preparation of Compound A-40

[0895] Compound A-40d (69.2 mg, 0.049 mmol) was dissolved in THF (0.6 mL), methanol (1.2 mL) and distilled water (0.2 mL) at -20 ° C, and then lithium hydroxide hydrate (20.6 mg, 0.492 mmol) dissolved in distilled water (1 mL) was slowly added dropwise, and the mixture was stirred for 2.5 hours while the temperature was slowly raised from -20 ° C to -5 ° C. The reaction solution was stirred at 0 ° C for another 4 hours. 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 (3 mL) and distilled water (3 mL), and then purified by preparative HPLC and freeze-dried to obtain compound A-40 (45.5 mg, 75%) in the form of a white solid; EI-MS m / z: 1224 [M+H] + .

[0896] Example I-33: Preparation of Compound A-41

[0897]

[0898] Step 1: Preparation of compound A-41a

[0899] Compound A-40c (37.7 mg, 0.047 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (33.8 mg, 0.047 mmol), HOBt (9.6 mg, 0.071 mmol), DIPEA (20.5 μL, 0.118 mmol) and pyridine (1 mL) 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), and washed by adding saturated sodium chloride 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-41a (48 mg, 74%) in the form of a white gum; EI-MS m / z: 1379 [M+H] + .

[0900] Step 2: Preparation of compound A-41

[0901] THF (0.35 mL) and methanol (0.87 mL) and distilled water (0.2 mL) were added to compound A-41a (48 mg, 0.035 mmol) at -20 ° C to dissolve the compound, and then lithium hydroxide hydrate (11 mg, 0.261 mmol) dissolved in distilled water (0.63 mL) was slowly added dropwise, and the mixture was stirred for 2.5 hours while the temperature was slowly raised from -20 ° C to -5 ° C. The reaction solution was stirred at 0 ° C for another 4.5 hours. 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 purified by preparative HPLC and freeze-dried to obtain compound A-41 (33.2 mg, 79%) in the form of a white solid; EI-MS m / z: 1210 [M+H] + .

[0902] Example I-34: Preparation of Compound A-42

[0903]

[0904] Step 1: Preparation of compound A-42b

[0905] Compound A-42a (Sigma Aldrich, CAS No.: 54881-49-1, 3.0 g, 17.6 mmol) was dissolved in MC (60.0 mL) at -78 ° C under a nitrogen atmosphere, and then a boron tribromide solution (1M-BBr3 in MC, 53.0 mL, 53.0 mmol) was slowly added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (500 mL) and distilled water (500 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-42b (1.8 g, 65%) in the form of a pink solid.

[0906] 1 H-NMR (400MHz, CDCl3) δ10.39 (s, 1H), 7.52 (dd, J = 7.2Hz, 2.0Hz, 1H), 7.35–7.27 (m, 2H), 5.81 (s, 1H).

[0907] Step 2: Preparation of compound A-42c

[0908] Compound A-42b (1.8 g, 11.5 mmol) was dissolved in ACN (60.0 mL) at 0 ° C under a nitrogen atmosphere, and potassium carbonate (3.98 g, 28.7 mmol) and benzyl bromide (2.05 mL, 1.72 mmol) were slowly added, and the mixture was stirred at 0 ° C for 10 minutes. The reaction temperature was raised to room temperature and the mixture was stirred for another 16 hours. After the reaction was completed, EA (200 mL) and distilled water (200 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-42c (2.0 g, 71%) in the form of a white solid.

[0909] 1 H-NMR (400MHz, CDCl3) δ10.54(s,1H),7.54(dd,J=7.6Hz,1.2Hz,1H),7.48–7.27(m,5H),7.19(dd,J=7.6Hz,J=1.2Hz,1H),5.21(s,2H).

[0910] Step 3: Preparation of compound A-42d

[0911] Compound A-42c (650.0 mg, 2.63 mmol) was dissolved in DMF (25.0 mL) under a nitrogen atmosphere, and then methyl thioglycolate (Merck, CAS No.: 2365-48-2, 0.47 mL, 5.27 mmol) and potassium carbonate (728.0 mg, 5.27 mmol) were added in sequence, and the mixture was stirred at room temperature for 30 minutes. The reaction temperature was raised to 80 ° C, and the mixture was stirred for another 3 hours. After the reaction was completed, EA (250 mL) and distilled water (250 mL) were added to extract the organic layer. The obtained organic layer was washed by adding a saturated sodium chloride aqueous solution (250 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain compound A-42d in the form of a yellow solid, which was used for the next step reaction without additional purification (230.0 mg, 29%).

[0912] Step 4: Preparation of Compound A-42e

[0913] Compound A-42d (400.0 mg, 1.34 mmol) was dissolved in MC (30.0 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (0.71 mL, 8.04 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 8.04 mL, 8.04 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 2 hours. After the reaction was completed, cooled distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (150 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-42e (125.0 mg, 28%) in the form of a white solid.

[0914] 1 H-NMR (400MHz, CDCl3) δ10.08(s,1H),9.01(s,1H),7.84(d,J=8.0Hz,1H),7.53-7.35(m,5H),7.04(d,J=8.0Hz,1H),5.38(s,2H),3.97(s,3H).

[0915] Step 5: Preparation of Compound A-42f

[0916] Compound A-42e (400.0 mg, 1.34 mmol) was dissolved in MC (10.0 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 1.14 ml, 1.14 mmol) was slowly added, and the mixture was then insulated and stirred for 1 hour. After the reaction was completed, distilled water (50.0 mL) was slowly added dropwise at -50 ° C to terminate the reaction, and EA (100 mL) and distilled water (50 mL) were added at room temperature 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-42f (40.0 mg, 44%) in the form of a white solid.

[0917] 1 H-NMR (400MHz, CDCl3) δ10.09(s,1H),9.03(s,1H),7.81(d,J=8.0Hz,1H),6.96(d,J=8.0Hz,1H),5.93(s,1H),3.98(s,3H).

[0918] Step 6: Preparation of Compound A-42g

[0919] Compound A-42f (40.0 mg, 0.169 mmol) was dissolved in ACN (10.0 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (Merck, 83.0 mg, 0.203 mmol) and silver (I) oxide (98.0 mg, 0.423 mmol) were added in sequence, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was diluted with EA (50 mL), filtered with diatomaceous earth, and washed by adding saturated sodium chloride aqueous solution (50 mL) to the solution. The reaction solution was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue obtained was obtained by column chromatography as a white solid compound A-42g (60.0 mg, 62%).

[0920] 1 H-NMR (400MHz, CDCl3) δ10.14(s,1H),9.00(s,1H),7.86(d,J=8.0Hz,1H),7.21(d,J=8.0Hz,1H),5.66(dd,J=10.4Hz,J=8.0Hz,1H),5.51(d, 3.2Hz,1H),5.29(d,8.0Hz,1H),5.17(dd,J=10.4Hz,J=3.2Hz,1H),4.29–4.19(m,3H),2.22(s,3H),2.108(s,3H),2.100(s,3H),2.04(s,3H).

[0921] Step 7: Preparation of Compound A-42h

[0922] Compound A-42g (59 mg, 0.104 mmol) was dissolved in THF (3.0 mL) at room temperature under a nitrogen atmosphere, and propargyl bromide (14.0 μL, 0.156 mmol), 1,2-diiodoethane (29.3 mg, 0.104 mmol) and zinc (zinc powder, 34.0 mg, 0.52 mmol) were added in sequence, and then ultrasonicated for 2 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and filtered with diatomaceous earth. The obtained filtrate was concentrated under reduced pressure. The obtained residue obtained by column chromatography gave compound A-42h (55.0 mg, 87%) in the form of a white solid.

[0923] 1H-NMR (400MHz, CDCl3) δ8.30 (d, J=9.6Hz, 1H), 7.44 (t, J=9.6Hz, 1H), 7.11 (dd, J=8.0Hz, J= 4.0Hz,1H),5.61(dd,J=10.4Hz,J=8.0Hz,1H),5.49(d,J=3.2Hz,1H),5.34–5.29(m,1H),5. 16–5.11(m,2H),4.29–4.25(m,1H),4.21–4.17(m,1H),4.14–4.09(m,1H),3.95(s,3H),2.7 7–2.75(m,2H),2.51(t,J=4.0Hz,1H),2.21(s,3H),2.13(s,3H),2.08(s,3H),2.03(s,3H).

[0924] Step 8: Preparation of compound A-42i

[0925] Compound A-42h (55.0 mg, 0.090 mmol) was dissolved in MC (3.0 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (27.5 mg, 0.136 mmol) and DIPEA (15.7 μL, 0.090 mmol) and pyridine (22.0 μL, 0.27 mmol) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, EA (50 mL) and distilled water (50 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-42i (32.0 mg, 45%) in the form of a white solid.

[0926] 1 H-NMR (400MHz, CDCl3) δ8.29(s,1H),8.25(d,J=9.2Hz,2H),7.47(d,J=8.0Hz,1H),7.34(d,J=9.2 Hz,2H),7.13(d,J=8.0Hz,1H),5.63(dd,J=10.4Hz,J=8.0Hz,1H),5.50(d,J=3.2Hz,1H),5.20–5. 12(m,2H),4.28–4.25(m,1H),4.21–4.16(m,1H),4.14–4.11(m,1H),3.96(s,3H),3.10–3.04(m,1 H),2.96–2.91(m,1H),2.21(s,3H),2.12(s,3H),2.08(s,3H),2.06(t,J=4.0Hz,1H),2.04(s,3H).

[0927] Step 9: Preparation of Compound A-42j

[0928] Compound A-42i (32.0 mg, 0.041 mmol) was dissolved in DMF (1.0 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (29.7 mg, 0.041 mmol), HOBt (8.3 mg, 0.061 mmol), DIPEA (18.0 μL, 0.102 mmol) and pyridine (1.0 mL) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, it was extracted with distilled water (10 mL), 2N-hydrochloric acid aqueous solution (5 mL) and EA (15 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 compound A-42j (31.0 mg, 55%) in the form of a white solid; EI-MS m / z: 1351 [M+H] + .

[0929] Step 10: Preparation of Compound A-42

[0930] Compound A-42j (30.0 mg, 0.022 mmol) was dissolved in methanol (1.0 mL) at 0°C under a nitrogen atmosphere, and then lithium hydroxide (7.0 mg, 0.166 mmol) dissolved in distilled water (1.0 mL) was slowly added dropwise, and the mixture was stirred at room temperature 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 purified by preparative HPLC and freeze-dried to obtain compound A-42 (14.8 mg, 68%) in the form of a white solid; EI-MS m / z: 1169 [M+H] + .

[0931] Example I-35: Preparation of Compound A-43

[0932]

[0933] Step 1: Preparation of compound A-43a

[0934] Selenium powder (640.0 mg, 8.10 mmol) was added to THF (27.0 mL) at room temperature under a nitrogen atmosphere, the mixture was cooled to 0 ° C, and n-butyl lithium solution (2.5 M n-BuLi in hexane, 3.56 mL, 8.91 mmol) was slowly added dropwise. The mixture was stirred at 0 ° C for 40 minutes, compound A-42c (2.0 g, 8.10 mmol) was dissolved in DMF (5.4 mL) and added, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, distilled water (100 mL) was slowly added dropwise to terminate the reaction. EA (100 mL) was added to the reaction solution, and the organic layer was then 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 A-43a (2.03 g, 72%) in the form of a yellow liquid.

[0935] 1 H-NMR (400MHz, CDCl3) δ10.71(s,1H),7.54(d,J=8.0Hz,1H),7.50–7.48(m,2H),7.42–7.32(m,4H),7.14(d,J =8.0Hz,1H),5.20(s,2H),2.89(t,J=7.6Hz,2H),1.71–1.50(m,2H),1.39–1.31(m,2H),0.81(t,J=7.6Hz,3H).

[0936] Step 2: Preparation of Compound A-43b

[0937] Compound A-43a (2.0 g, 5.75 mmol) was dissolved in DMF (25.0 mL) under a nitrogen atmosphere, and then ethyl bromoacetate (1.6 mL, 13.4 mmol) and potassium carbonate (1.85 g, 13.4 mmol) were added in sequence, and the mixture was stirred at 120 ° C for 16 hours. After the reaction was completed, distilled water (250 mL) and EA (250 mL) were added to extract the organic layer. The obtained organic layer was washed by adding a saturated aqueous sodium chloride solution (250 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was obtained by column chromatography as a white solid compound A-43b (1.5 g, 69%).

[0938] 1H-NMR (400MHz, CDCl3) δ10.63 (s, 1H), 7.56 (d, J = 8.0Hz, 1H), 7.50–7.49 (m, 2H), 7.44–7.39 (m, 3H), 7.37–7 .33(m,1H),7.18(d,J=8.0Hz,1H),5.22(s,1H),3.95(q,J=7.2Hz,2H),3.53(s,2H),1.05(t,J=7.2Hz,3H).

[0939] Step 3: Preparation of compound A-43c

[0940] Compound A-43b (1.5 g, 3.97 mmol) was dissolved in DMF (20.0 mL) at 0 ° C under a nitrogen atmosphere, and potassium carbonate (824.0 mg, 5.96 mmol) was then added, and the mixture was stirred at 120 ° C for 2 hours. After the reaction was completed, EA (200 mL) and distilled water (200 mL) were added to extract the organic layer. The obtained organic layer was washed by adding a saturated sodium chloride aqueous solution (200 mL). The 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-43c (1.2 g, 84%) in the form of a white solid.

[0941] 1 H-NMR (400MHz, CDCl3) δ8.30 (s, 1H), 7.51 (d, J = 8.0Hz, 1H), 7.48–7.46 (m, 2H), 7.42–7.38 (m, 2H), 7.3 5–7.32(m,2H),6.87(d,J=8.0Hz,1H),5.27(s,2H),4.38(q,J=7.2Hz,2H),4.38(t,J=7.2Hz,3H); EI-MS m / z:360[M+H] + .

[0942] Step 4: Preparation of Compound A-43d

[0943] Compound A-43c (1.2 g, 3.34 mmol) was dissolved in MC (30.0 mL) at 0 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (1.19 mL, 13.36 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 13.36 mL, 13.36 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 3 hours. After the reaction was completed, cooled distilled water (150 mL) was slowly added dropwise to terminate the reaction, and EA (150 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-43d (1.09 g, 89%) in the form of a white solid.

[0944] 1 H-NMR (400MHz, CDCl3) δ10.04(s,1H),9.32(s,1H),7.84(d,J=8.0Hz,1H),7.48–7.35(m ,5H),7.00(d,J=8.0Hz,1H),5.37(s,2H),4.41(q,J=7.2Hz,2H),1.42(t,J=7.2Hz,3H).

[0945] Step 5: Preparation of Compound A-43e

[0946] Compound A-43d (420.0 mg, 1.11 mmol) was dissolved in MC (20.0 mL) at -78 ° C under nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 3.33 ml, 3.33 mmol) was slowly added, and the mixture was then insulated and stirred for 2 hours. After the reaction was completed, distilled water (50 mL) was slowly added dropwise at -50 ° C to terminate the reaction, and EA (150 mL) and distilled water (100 mL) were added at room temperature 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-43e (300.0 mg, 90%) in the form of a white solid.

[0947] 1 H-NMR (400MHz, CDCl3) δ10.05 (s, 1H), 9.33 (s, 1H), 7.81 (d, J = 8.0Hz, 1H), 6.9 2(d,J=8.0Hz,1H),5.98(s,1H),4.41(q,J=7.2Hz,2H),1.43(t,J=7.2Hz,3H).

[0948] Step 6: Preparation of Compound A-43f

[0949] Compound A-43e (150.0 mg, 0.50 mmol) was dissolved in ACN (30.0 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (Merck, 250.0 mg, 0.60 mmol) and silver (I) oxide (290.0 mg, 1.25 mmol) were added in sequence, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was diluted with EA (50 mL), filtered with diatomaceous earth, and washed by adding saturated sodium chloride aqueous solution (80 mL) to the solution. The reaction solution was dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue obtained was obtained by column chromatography as a white solid compound A-43f (290.0 mg, 91%).

[0950] 1 H-NMR (400MHz, CDCl3) δ10.11(s,1H),9.30(s,1H),7.87(d,J=8.0Hz,1H),7.15(d, J=8.0Hz,1H),5.63(dd,J=10.4Hz,8.0Hz,1H),7.52(d,J=3.2Hz,1H),5.28(d,J=8. 0Hz,1H),5.17(dd,J=10.4Hz,3.2Hz,1H),4.42(d,J=7.2Hz,1H),4.31–4.26(m,1H) ,4.22–4.14(m,2H),2.22(s,3H),2.10(s,6H),2.04(s,3H),1.43(t,J=7.2Hz,3H).

[0951] Step 7: Preparation of Compound A-43g

[0952] Compound A-43f (52.0 mg, 0.082 mmol) was dissolved in THF (3.0 mL) at room temperature under a nitrogen atmosphere, and propargyl bromide (11.1 μL, 0.124 mmol), 1,2-diiodoethane (23.0 mg, 0.082 mmol) and zinc (zinc powder, 27.0 mg, 0.414 mmol) were added in sequence, and then ultrasonicated for 3 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and filtered with diatomaceous earth. The filtrate was concentrated under reduced pressure. The resulting residue obtained was subjected to column chromatography to give compound A-43g (24.0 mg, 44%) as a white solid.

[0953] 1H-NMR (400MHz, CDCl3) δ8.55 (d, J=9.2Hz, 1H), 7.51–7.46 (m, 1H), 7.07–7.03 (m, 1H), 5.60 (dd, J=10.4Hz,8.0Hz,1H),5.49(d,J=3.2Hz,1H),5.34–5.29(m,1H),5.17–5.12(m,2H),4.40(q,J= 7.2Hz,2H),4.29–4.25(m,1H),4.21–4.17(m,1H),4.14–4.09(m,1H),2.77–2.75(m,2H),2.50– 2.48(m,1H),2.21(s,3H),2.12(s,3H),2.09(s,3H),2.03(s,3H),1.42(t,J=7.2Hz,3H); EI-MS m / z:690[M+Na] + .

[0954] Step 8: Preparation of Compound A-43h

[0955] Compound A-43g (22.0 mg, 0.033 mmol) was dissolved in MC (3.0 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (10.0 mg, 0.049 mmol), DIPEA (15.0 μL, 0.085 mmol) and pyridine (8.0 μL, 0.099 mmol) were added in sequence, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, EA (50 mL) and distilled water (50 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-43h (10.0 mg, 37%) in the form of a white solid.

[0956] 1H-NMR (400MHz, CDCl3) δ8.54(s,1H),8.25(d,J=9.2Hz,2H),7.51–7.49(m,1H),7.35(d,J=9.2Hz,2H),7.0 8–7.06(m,1H),6.25(t,J=7.2Hz,2H),5.60(dd,J=10.4Hz,J=8.0Hz,1H),5.50(d,J=3.2Hz,1H),5.20–5.1 2(m,2H),4.41(t,J=7.2Hz,2H),4.30–4.25(m,1H),4.21–4.09(m,2H),3.08–3.02(m,1H),2.96–2.90(m,1 EI-MS m / z:855[M+Na] + .

[0957] Step 9: Preparation of Compound A-43i

[0958] Compound A-43h (10.0 mg, 0.012 mmol) was dissolved in DMF (1.0 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (8.6 mg, 0.012 mmol), HOBt (2.43 mg, 0.018 mmol), DIPEA (5.23 μL, 0.03 mmol) and pyridine (1.0 mL) were added in sequence, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, it was extracted with distilled water (20 mL), 2N-hydrochloric acid aqueous solution (5 mL) and EA (25 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 compound A-43i (10.0 mg, 58%) in the form of a white solid; EI-MS m / z: 1412 [M+H] + .

[0959] Step 10: Preparation of Compound A-43

[0960] Compound A-43i (10.0 mg, 0.007 mmol) was dissolved in methanol (1.0 mL) at 0 ° C under a nitrogen atmosphere, and then lithium hydroxide (2.23 mg, 0.053 mmol) dissolved in distilled water (1.0 mL) was slowly added dropwise, and the mixture was stirred at room temperature for 5 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 purified by preparative HPLC and freeze-dried to obtain compound A-43 (5.2 mg, 60%) in the form of a white solid; EI-MS m / z: 1216 [M+H] + .

[0961] Example I-36: Preparation of Compound A-44

[0962]

[0963] Step 1: Preparation of compound A-44a

[0964] Compound C-8b (8.5 g, 33.69 mmol) was dissolved in MC (168 mL) at room temperature under nitrogen atmosphere, (carboethoxymethylene)triphenylphosphorane (combi-blocks, CAS No.: 1099-45-2, 23.5 g, 67.46 mmol) was added, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-44a (9.96 g, 92%) in the form of a white solid.

[0965] 1 H-NMR (400MHz, CDCl3) δ7.53–7.33(m,6H),7.28–7.24(m,1H),7.11–7.07(m,2H),6.71(d,J=8H z,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] + .

[0966] Step 2: Preparation of Compound A-44b

[0967] Compound A-44a (9.96 g, 30.9 mmol) was dissolved in THF (250 mL) and methanol (750 mL) at room temperature under a nitrogen atmosphere, and then 5% palladium on carbon (5% Pd / C, 5.1 g) was added, and the mixture was stirred for 2 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered with diatomaceous earth. The filtered solution was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to obtain compound A-44b (7 g, 97%) as a white solid.

[0968] 1 H-NMR (400MHz, CDCl3) δ7.10–7.01(m,2H),6.59(d,J=7.6Hz,1H),6.49(s,1H),5.05(s,1H),4. 16(q,J=7.2Hz,2H),3.10(t,J=7.6Hz,2H),2.75(t,J=7.6Hz,2H),1.26(t,J=7.2Hz,3H); EI-MS m / z:235[M+H] + .

[0969] Step 3: Preparation of Compound A-44c

[0970] Compound A-44b (7 g, 29.88 mmol) was dissolved in ACN (230 mL) at room temperature under a nitrogen atmosphere, and potassium carbonate (10.3 g, 74.7 mmol) and benzyl bromide (4.6 mL, 38.85 mmol) were then added, and the mixture was stirred at 80 ° C for 2.5 hours. After the reaction was completed, EA (1 L), distilled water (500 mL) and 2N-hydrochloric acid aqueous solution (200 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 A-44c (9.63 g, 99%) in the form of a colorless oil.

[0971] 1 H-NMR (400MHz, CDCl3) δ7.49–7.44(m,2H),7.42–7.36(m,2H),7.36–7.30(m,1H),7.12(t,J=8Hz,1H),7.05(d,J=8Hz,1H),6.68(d,J= 8Hz,1H),6.57(s,1H),5.18(s,2H),4.16(q,J=7.2Hz,2H),3.10(t,J=7.6Hz,2H),2.74(t,J=7.6Hz,2H),1.25(t,J=7.2Hz,3H); EI-MS m / z:325[M+H] + .

[0972] Step 4: Preparation of Compound A-44d

[0973] Compound A-44c (9.63 g, 29.69 mmol) was dissolved in MC (600 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (8 mL, 89.06 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 89 mL, 89.06 mmol) were slowly added in sequence, and the mixture was insulated and stirred for 2 hours. After the reaction was completed, distilled water (800 mL) was slowly added dropwise to terminate the reaction, and MC (400 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-44d (5.1 g, 49%) in the form of a yellow solid.

[0974] 1 H-NMR (400MHz, CDCl3) δ10.25(s,1H),7.68(d,J=8.4Hz,1H),7.48–7.33(m,5H),6.80(d,J=8.4Hz,1H),6.63(s ,1H),5.26(s,2H),4.17(q,J=7.2Hz,2H),3.18(t,J=7.6Hz,2H),2.78(t,J=7.6Hz,2H),1.25(t,J=7.2Hz,3H).

[0975] Step 5: Preparation of Compound A-44e

[0976] Compound A-44d (1.5 g, 4.26 mmol) was dissolved in MC (85 mL) at -78 ° C under a nitrogen atmosphere, and then pentamethylbenzene (Alfa aesar, CAS No.: 700-12-9, 1.89 g, 12.74 mmol) and boron trichloride solution (1M-BCl3 in MC, 12.8 mL, 12.8 mmol) were slowly added, and the mixture was stirred at -78 ° C for 2 hours. After the reaction was completed, distilled water (160 mL) was slowly added dropwise to terminate the reaction, and 2N-sodium hydroxide aqueous solution was added at 0 ° C to neutralize the reaction solution. EA (200 mL) and distilled water (100 mL) were added to the mixture to extract the organic layer. The obtained organic layer was washed by adding saturated sodium chloride aqueous solution (100 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-44e (954 g, 85%) in the form of a beige solid.

[0977] 1H-NMR(400MHz,DMSO-d6)δ11.13(brs,1H),10.05(s,1H),7.61(d,J=8.4Hz,1H),6.75–6.71(m,2 H), 4.09 (q, J = 7.2Hz, 2H), 3.07 (t, J = 7.2Hz, 2H), 2.76 (t, J = 7.2Hz, 2H), 1.18 (t, J = 7.2Hz, 3H); MS m / z:263[M+H] + .

[0978] Step 6: Preparation of Compound A-44f

[0979] Compound A-44e (433mg, 1.65mmol) is dissolved in ACN (24mL) and THF (6mL) at 0°C under a nitrogen atmosphere, then acetyl bromide-α-D-glucuronic acid methyl ester (TCI, 787mg, 1.98mmol) is added, and the mixture is insulated and stirred for 20 minutes. In addition, silver oxide (I) (957mg, 4.13mmol) and molecular sieves (200mg) are added and stirred for 10 minutes, and then the mixture is stirred at room temperature for 2 hours. After the reaction is completed, the reaction solution is diluted with EA (200mL) and filtered with a diatomaceous earth filter, and then the solution is concentrated under reduced pressure. The obtained residue obtains compound A-44f (90mg, 9%) in the form of a brown solid through column chromatography.

[0980] 1 H-NMR (400MHz, CDCl3) δ10.31(s,1H),7.71(d,J=8.4Hz,1H),6.88(d,J=8.4Hz,1H),6.51(s,1H),5.40–5.35(m,4H),4.28–4.26( MS m / z:579[M+H] + .

[0981] Step 7: Preparation of Compound A-44g

[0982] Compound A-44f (88 mg, 0.15 mmol) was dissolved in THF (5 mL) at room temperature under a nitrogen atmosphere, and propargyl bromide (25.4 μL, 0.29 mmol), 1,2-diiodoethane (43 mg, 0.15 mmol) and zinc (zinc powder, 50 mg, 0.76 mmol) were added in sequence, and then ultrasonicated for 1.5 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and filtered with diatomaceous earth. Distilled water (100 mL) was added to the filtered solution, and the organic layer was extracted. The obtained organic layer was washed by adding a saturated sodium chloride aqueous solution (100 mL), then dried over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained residue was subjected to column chromatography to obtain compound A-44g (70 mg, 74%) in the form of a beige solid.

[0983] 1 H-NMR (400MHz, CDCl3) δ7.24–7.23 (m, 1H), 6.80 (d, J = 8.4Hz, 1H), 6.46 (s, 1H),5.36–5.35(m,3H),5.28–5.25(m,1H),5.19–5.18(m,1H),4.20–4.15(m ,3H),3.74(s,3H),3.11(t,J=7.6Hz,2H),2.88–2.77(m,2H),2.73(t,J=7.6 Hz,2H),2.52–2.49(m,1H),2.06(s,6H),2.05(s,3H),1.29–1.25(m,3H); MS m / z:640[M+Na] + .

[0984] Step 8: Preparation of Compound A-44h

[0985] Compound A-44g (67 mg, 0.11 mmol) was dissolved in MC (2 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrobenzene chloroformate (44 mg, 0.22 mmol), pyridine (26.2 μL, 0.33 mmol) and DIPEA (28.3 μL, 0.16 mmol) were added in sequence, and the mixture was stirred at 0 ° C for 30 minutes. After the reaction was completed, the reaction solution was diluted with EA (15 mL), 2N-hydrochloric acid aqueous solution (15 mL) was added, and the organic layer was extracted. The organic layer was washed by adding saturated sodium chloride aqueous solution (15 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-44h (74.4 mg, 88%) in the form of a white solid.

[0986] 1H-NMR (400MHz, CDCl3) δ8.26(d,J=8.8Hz,2H),7.37(d,J=8Hz,2H),7.27(m,1H),6.81(d,J=8.4Hz,1H),6.49(s,1H),6.25–6.22(m,1H),5.37–5.3 6(m,3H),5.24–5.23(m,1H),4.21–4.15(m,3H),3.74(s,3H),3.15–3.03 (m,4H),2.75(t,J=7.6Hz,2H),2.07–2.05(m,10H),1.29–1.25(m,3H); MS m / z:805[M+Na] + .

[0987] Step 9: Preparation of Compound A-44i

[0988] Compound A-44h (40 mg, 0.051 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (37 mg, 0.052 mmol), HOBt (10 mg, 0.074 mmol), DIPEA (22.2 μL, 0.128 mmol) and pyridine (1 mL) 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 (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 obtained residue was subjected to column chromatography to obtain compound A-44i (36 mg, 52%) in the form of a white solid; EI-MS m / z: 1363 [M + H] + .

[0989] Step 10: Preparation of Compound A-44

[0990] Compound A-44i (36 mg, 0.026 mmol) was dissolved in a mixture of methanol (0.8 mL), distilled water (0.2 mL) and THF (0.15 mL) at -20 ° C, and then lithium hydroxide hydrate (11.1 mg, 0.264 mmol) dissolved in distilled water (0.8 mL) was added dropwise, and the mixture was stirred at -20 ° C for 1.5 hours and at room temperature for 5.5 hours. 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 purified by preparative HPLC and freeze-dried to obtain compound A-44 (17.8 mg, 56%) in the form of a white solid; EI-MS m / z: 1195 [M+H] + .

[0991] Example I-37: Preparation of Compound A-45

[0992]

[0993] Step 1: Preparation of compound A-45a

[0994] Compound A-44h (45 mg, 0.057 mmol) was dissolved in DMF (1 mL) at 0 ° C under a nitrogen atmosphere, and then MMAF-OMe (CAS No.: 863971-12-4, 43 mg, 0.058 mmol), HOBt (12 mg, 0.089 mmol), DIPEA (25 μL, 0.144 mmol) and pyridine (1 mL) 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 (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 obtained residue was subjected to column chromatography to give A-45a (47.7 mg, 60%) in the form of a white solid; EI-MS m / z: 1391 [M+H] + .

[0995] Step 2: Preparation of Compound A-45

[0996] Compound A-45a (47 mg, 0.034 mmol) was dissolved in a mixture of methanol (0.9 mL), distilled water (0.1 mL) and THF (0.17 mL) at -20°C, and then lithium hydroxide hydrate (14.2 mg, 0.338 mmol) dissolved in distilled water (0.8 mL) was added dropwise, and the mixture was stirred at -20°C for 2 hours, and then stirred for 5.5 hours while slowly warming to 0°C. 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 purified by preparative HPLC and freeze-dried to obtain compound A-45 (25.4 mg, 62%) in the form of a white solid; EI-MS m / z: 1209 [M+H] + .

[0997] Example I-38: Preparation of Compounds A-46, A-47 and A-48

[0998]

[0999] Step 1: Preparation of compound A-46a

[1000] Core C-10 (2.0 g, 6.46 mmol) was dissolved in MC (100.0 mL) at -78 ° C under a nitrogen atmosphere, and then dichloromethyl methyl ether (1.73 mL, 19.4 mmol) and titanium tetrachloride solution (1M-TiCl4 in MC, 19.4 mL, 19.4 mmol) were slowly added in sequence, and the mixture was stirred for 2 hours. After the reaction was completed, cooled distilled water (100 mL) was slowly added dropwise to terminate the reaction, and EA (300 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-46a (1.32 g, 60%) in the form of a white solid.

[1001] 1 H-NMR (400MHz, CDCl3) δ10.04(s,1H),8.04(s,1H),7.56(d,J=8.0Hz,1H),7.48–7.36 (m,5H),6.87(d,J=8.0Hz,1H),5.29(s,2H),4.39–4.34(m,5H),1.41(t,J=7.2Hz,3H).

[1002] Step 2: Preparation of Compound A-46b

[1003] Compound A-46a (1.32 g, 3.91 mmol) was dissolved in MC (80.0 mL) at -78 ° C under a nitrogen atmosphere, and then a boron trichloride solution (1M-BCl3 in MC, 11.7 mL, 11.7 mmol) was slowly added, and the mixture was then insulated and stirred for 3 hours. After the reaction was completed, distilled water (100.0 mL) was slowly added dropwise at -50 ° C to terminate the reaction, and EA (300 mL) and distilled water (200 mL) were added at room temperature 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-46b (550.0 g, 56%) in the form of an ivory solid.

[1004] 1 H-NMR (400MHz, CDCl3) δ10.03(s,1H),8.03(s,1H),7.49(d,J=8.0Hz,1H),6.68(d,J =8.0Hz,1H),5.62(s,1H),4.43(s,1H),4.38(q,J=7.2Hz,2H),1.42(t,J=7.2Hz,3H).

[1005] Step 3: Preparation of Compound A-46c

[1006] Compound A-46b (550.0 mg, 2.23 mmol) was dissolved in MC (50.0 mL) at 0 ° C under a nitrogen atmosphere, and then acetyl bromide-α-D-galactose (Merck, 1.0 g, 2.45 mmol) and benzyltributylammonium chloride (694.0, 2.23 mmol) were added. 5N sodium hydroxide aqueous solution (1.33 mL, 6.69 mmol) was slowly added to the reaction solution, and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, MC (150 mL) and distilled water (150 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-46c (910.0 mg, 70%) in the form of an ivory solid.

[1007] 1 H-NMR (400MHz, CDCl3) δ10.10 (s, 1H), 8.03 (s, 1H), 7.55 (d, J = 8.0Hz, 1H), 6.96 ( d,J=8.0Hz,1H),5.66(dd,J=10.4Hz,J=8.0Hz,1H),5.51(d,J=3.2Hz,1H),5.39(d J=9.2Hz,1H)5.18(dd,J=10.4Hz,J=3.2Hz,1H),4.38(t,J=7.2Hz,2H),4.30(s,3H),4.27–4.20(m, 1H),4.19–4.16(m,2H),2.20(s,3H),2.08(s,3H),2.07(s,3H),2.03(s,3H),1.42(t,J=7.2Hz,3H).

[1008] Step 4: Preparation of compound A-46d

[1009] Compound A-46c (900.0 mg, 1.56 mmol) was dissolved in THF (40.0 mL) at room temperature under a nitrogen atmosphere, and propargyl bromide (0.2 mL, 2.33 mmol), 1,2-diiodoethane (440.0 mg, 1.56 mmol) and zinc (zinc powder, 508.0 mg, 7.79 mmol) were added in sequence, and then ultrasonicated for 6 hours, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was diluted with EA (100 mL) and filtered with diatomaceous earth. The filtrate was concentrated under reduced pressure. The resulting residue was subjected to column chromatography to give compound A-46d (825.0 mg, 85%) in the form of an ivory solid.

[1010] 1H-NMR (400MHz, CDCl3) δ7.38(d,J=9.2Hz,1H),7.08(t,J=9.2Hz,1H),6.88–6.85(m,1H),5.62( dd,J=10.4Hz,J=8.0Hz,1H),5.48(d,J=3.2Hz,1H),5.26–5.20(m,2H),5.15(dd,J=10.4Hz,J=3. 2Hz,2H),4.36(q,J=7.2Hz,2H),4.26(s,3H)4.24–4.09(m,3H),2.78–2.77(m,2H),2.41(t,J=3 .2Hz,1H),2.21(s,3H),2.06(s,6H),2.02(s,3H),1.41(t,J=7.2Hz,3H),1.25(t,J=7.2Hz,1H).

[1011] Step 5: Preparation of Compound A-46e

[1012] Compound A-46d (100.0 mg, 0.162 mmol) was dissolved in MC (10.0 mL) at 0 ° C under a nitrogen atmosphere, and then 4-nitrophenyl chloroformate (49.0 mg, 0.243 mmol), DIPEA (70.0 μL, 0.405 mmol) and pyridine (40.0 μL, 0.486 mmol) were added in sequence, and the mixture was stirred for 3 hours while the temperature was slowly raised from 0 ° C to room temperature. After the reaction was completed, EA (50 mL) and distilled water (50 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-46e (56.0 mg, 44%) in the form of a white solid.

[1013] 1H-NMR (400MHz, CDCl3) δ8.24(d,J=9.2Hz,2H),7.38(s,1H),7.34(d,J=9.2Hz,2H),7.11(d,J=8.0Hz,2H) ,6.88(d,J=8.0Hz,1H),6.14(t,J=7.2Hz,1H),5.63(dd,J=10.4Hz,J=8.0Hz,1H),5.49(d,J=3.2Hz,1H), 5.29–5.23(m,1H),5.15(dd,J=10.4Hz,J=3.2Hz,1H),4.44(t,J=7.2Hz,2H),4.27(s,3H),4.24–4.09(m, 3H),3.09–3.04(m,1H),2.97–2.90(m,1H),2.21(s,3H),2.06(s,6H),2.01(s,3H),1.42(t,J=7.2Hz,3H).

[1014] Step 6: Preparation of Compound A-46f

[1015] Compound A-46e (56.0 mg, 0.071 mmol) was dissolved in DMF (1.0 mL) at 0 ° C under a nitrogen atmosphere, and then MMAE (51.3 mg, 0.071 mmol), HOBt (14.5 mg, 0.107 mmol), DIPEA (31 μL, 0.178 mmol) and pyridine (1.0 mL) were added in sequence, and the mixture was stirred at room temperature for 3 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 purified by preparative HPLC and freeze-dried to obtain compound A-46f (4.0 mg, 4.1%) in the form of a white solid; EI-MS m / z: 1361 [M+H] + .

[1016] Step 7: Preparation of compounds A-46, A-47 and A-48

[1017] Compound A-46f (4.0 mg, 0.0014 mmol) was dissolved in methanol (0.5 mL) at 0°C under a nitrogen atmosphere, and then lithium hydroxide (0.5 mg, 0.011 mmol) dissolved in distilled water (0.5 mL) 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 purified by preparative HPLC, and freeze-dried to obtain compounds A-46 (acid form), A-47 (methyl ester form) and A-48 (ethyl ester form) in the form of white solids, respectively.

[1018] (A-46:0.5mg, 14%; EI-MS m / z:1166[M+H] + ), (A-47:0.4mg, 11%; MS m / z:1179[M+H] + ), (A-48: 1.0mg, 28%; MS m / z: 1193[M+H] + ).

[1019] Example I-39: Preparation of Compound A-49

[1020]

[1021] Compound A-36 (2.0 mg, 0.0015 mmol) was dissolved in DMSO (1.5 mL) at 0 ° C under a nitrogen atmosphere, and then a solution of linker P-5 (1.0 mg) dissolved in DMSO (500 μL) was added to the reaction. In addition, distilled water (2 mL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (1 mg) were added to the reaction solution in sequence, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was diluted with distilled water (3 mL), purified by preparative HPLC, and then freeze-dried to obtain compound A-49 (1.7 mg, 69%) in the form of a white solid; MS m / z: 1549 [M+H] + .

[1022] Example I-40: Preparation of Compound A-50

[1023]

[1024] Compound A-50a was obtained in a similar manner to Example I-39, except that linker P-6 was used instead of linker P-5 in Example I-39. Then, compound A-50a was subjected to Boc deprotection to obtain compound A-50 (1.2 mg, 51%); MS m / z: 1492 [M+H] + .

[1025] Example I-41: Preparation of Compound A-51

[1026]

[1027] In a similar manner to Example I-39, compound A-51 (1.5 mg, 63%; EI-MS m / z: 1502 [M+H]) was obtained as a white solid. + ), except that compound P-3c is used instead of linker P-5 in Example I-39.

[1028] Example I-42: Preparation of Compound A-52

[1029]

[1030] Compound A-52 (1.2 mg, 51%; EI-MS m / z: 1476 [M+H]) was obtained in a similar manner to Example I-39 as a white solid. + ), except that compound P-3 is used instead of linker P-5 in Example I-39.

[1031] Example II-1: Preparation of Compound B-1

[1032]

[1033] Compound A-15 (3.1 mg, 0.0026 mmol) was dissolved in DMSO (200 μL) at 0 ° C under a nitrogen atmosphere, and then 300 μL of a solution of linker Q-1 (5 mg) dissolved in DMSO (1 mL) was added to the reaction. In addition, distilled water (2 mL), ethanol (20 μL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (1 mg) were added to the reaction solution in sequence, and the mixture was stirred at room temperature for 20 minutes. After the reaction was completed, the reaction solution was diluted with distilled water (1 mL), purified by preparative HPLC, and then freeze-dried to obtain compound B-1 (1.9 mg, 48.8%) in the form of a white solid; MS m / z: 1465 [M+H] + .

[1034] Example II-2: Preparation of Compound B-2

[1035]

[1036] Compound A-11 (3.0 mg, 0.0025 mmol) was dissolved in DMSO (200 μL) at 0 ° C under a nitrogen atmosphere, and then 150 μL of a solution of linker Q-1 (7.5 mg) dissolved in DMSO (1 mL) was added to the reaction. In addition, distilled water (2 mL), ethanol (20 μL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (1 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), purified by preparative HPLC, and then freeze-dried to obtain compound B-2 (2.9 mg, 77.3%) in the form of a white solid; MS m / z: 1481 [M+H] + .

[1037] Example II-3: Preparation of Compound B-3

[1038]

[1039] Compound A-16 (3.9 mg, 0.0032 mmol) was dissolved in DMSO (200 μL) at 0 ° C under a nitrogen atmosphere, and then 150 μL of a solution of linker Q-1 (18 mg) dissolved in DMSO (1.8 mL) was added to the reaction. In addition, distilled water (2 mL), ethanol (20 μL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (1 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), purified by preparative HPLC, and then freeze-dried to obtain compound B-3 (2.9 mg, 59.6%) in the form of a white solid; MS m / z: 1507 [M+H] + .

[1040] Example II-4: Preparation of Compound B-4

[1041]

[1042] Compound A-12 (3.6 mg, 0.0029 mmol) was dissolved in DMSO (200 μL) at 0 ° C under a nitrogen atmosphere, and then 150 μL of a solution of linker Q-1 (14 mg) dissolved in DMSO (1 mL) was added to the reaction. In addition, distilled water (2 mL), ethanol (20 μL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (1 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), purified by preparative HPLC, and then freeze-dried to obtain compound B-4 (3.0 mg, 66.9%) in the form of a white solid; MS m / z: 1508 [M+H] + .

[1043] Example II-5: Preparation of Compound B-5

[1044]

[1045] Compound A-17 (9 mg, 0.0073 mmol) was dissolved in DMSO (200 μL) at 0°C under a nitrogen atmosphere, and then 320 μL of a solution of linker Q-1 (18 mg) dissolved in DMSO (1.8 mL) was added to the reaction. In addition, distilled water (2 mL), ethanol (20 μL), copper sulfate (II) pentahydrate (CuSO4·5H2O, 1 mg) and sodium ascorbate (2 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), purified by preparative HPLC, and then freeze-dried to obtain compound B-5 (4.4 mg, 40%) in the form of a white solid; MS m / z: 1528 [M+H] + .

[1046] Example II-6: Preparation of Compound B-6

[1047]

[1048] Compound A-12 (4.2 mg, 0.0034 mmol) was dissolved in DMSO (200 μL) at 0 ° C under a nitrogen atmosphere, and then 550 μL of a solution of connector Q-2 (12 mg) dissolved in DMSO (1 mL) was added to the reaction. In addition, distilled water (2 mL), DMSO (400 μl), copper sulfate (II) pentahydrate (CuSO4 · 5H2O, 1 mg) and sodium ascorbate (1.4 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) and ACN (1 mL), purified by preparative HPLC, and then freeze-dried to obtain compound B-6 (3.1 mg, 66%) in the form of a white solid; EI-MS m / z: 1400 1 / 2 [M+H] + ,934 1 / 3[M+H] + .

[1049] Example II-7: Preparation of Compound B-7

[1050]

[1051] Compound A-17 (5 mg, 0.004 mmol) was dissolved in DMSO (1.5 mL) and ethanol (200 μL) at 0 ° C under a nitrogen atmosphere, and then 100 μL of a solution of linker Q-2 (12 mg) dissolved in DMSO (1.2 mL) was added to the reaction. In addition, distilled water (2 mL), ethanol (20 μL), copper (II) sulfate pentahydrate (CuSO4·5H2O, 1 m...

Claims

1. A linker compound represented by the following formula 1, or a pharmaceutically acceptable salt thereof: [Formula 1] A—(L 1 ) k -U j in, 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, PL is an active agent which is bonded to L via a heteroatom selected from N, O and S. 2 or with benzene ring and A-(L 1 )k-bonded carbon atom connection; L 2 is a self-eliminating joint, which is selected so that L 2 With benzene ring and A-(L 1 )k-bonded carbon atoms to promote the cleavage of the bond between L 2 Cleavage of the bond with PL; W is an optional substituent on the benzene ring; Z 1 and Z 3 One of the following is selected from NR 3 , O, S and Se, and Z 1 and Z 3 Another one in Z 2 each independently represents CH or N, and -(V)h, if present, each independently replaces H in NH or CH; Indicates that A-(L 1 )k-formed bonds; R 3 is H or a C1-C8 hydrocarbon group; V is an electron-withdrawing group, an electron-donating group or -CH2-(L 2 )l-PL-; 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 linker compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The ring in formula A Selected from the following groups:

3. The linker compound according to claim 1, wherein V is selected from the group consisting of halogen, CN, NO2, formyl, C1-C8 alkylcarbonyl, carboxyl, C1-C8 alkoxycarbonyl, carbamoyl, mono-C1-C8 alkylcarbamoyl, di-C1-C8 alkylcarbamoyl, C1-C8 alkyl, C1-C8 alkenyl, OH, C1-C8 alkoxy, SH, C1-C8 alkylthiol, NH2, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, C6-C 18 Aryl, -CH2-PL and -CH2-L 2 -PL.

4. The linker compound according to claim 1, wherein W is selected from the group consisting of: H, C1-C 12 Saturated or unsaturated hydrocarbon group, halogen, halogenated-C1-C8 alkyl, CN, NO2, OH, C1-C8 alkoxy, hydroxy-C1-C8 alkyl, C1-C8 alkoxy-C1-C8 alkyl, SH, C1-C8 alkylthio, mercapto-C1-C8 alkyl, amino, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, amino-C1-C8 alkyl, C1-C8 monoalkylamino-C1-C8 alkyl, C1-C8 dialkylamino- C1-C8 alkyl, carboxyl, C1-C8 alkoxycarbonyl, C1-C8 alkoxycarbonyloxy, carboxyl-C1-C8 alkyl, C1-C8 alkoxycarbonyl-C1-C8 alkyl, carbamoyl, mono-C1-C8 alkylcarbamoyl, di-C1-C8 alkylcarbamoyl, carbamoyl-C1-C8 alkyl, mono-C1-C8 alkylcarbamoyl-C1-C8 alkyl and di-C1-C8 alkylcarbamoyl-C1-C8 alkyl.

5. The linker compound 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 Including the form in which the -OH group is 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 is OH, mono-C1-C8 alkylamino, di-C1-C8 alkylamino or -NH(CH2CH2O) f R t6 , where R t6 is H or C1-C4 alkyl; f is an integer of 1-10, and r and s are integers of 1-5 each.

6. The linker compound according to claim 1, wherein -(X) x -L 3 -for and R 8 and R 9 Each is independently selected from the group consisting of H, halogen, C1-C8 alkyl, CN and NO2, and o is an integer from 0 to 2.

7. The linker compound 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 linker compound 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 linker compound according to claim 1, wherein The PL is selected from the group consisting of:

10. The linker compound according to claim 1, wherein L 2 Selected from -OC(=O)-, -S(=O)2-, The group composed of R 10 To R 12 Each is independently H, C1-C8 alkyl, amino-C1-C8 alkyl, C1-C8 alkyl substituted with mono-(C1-C8 alkyl)amino or di-(C1-C8 alkyl)amino, or -(CH2CH2O) g R 13 ,in R 13 is H or a C1-C4 alkyl group, and g is an integer from 1 to 10.

11. The linker compound 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 linker compound according to claim 1, wherein The binding functional group includes a functional group selected from the group consisting of halogen, OH, C1-C8 alkoxy, hydroxyamino, COH, C1-C8 alkylcarbonyl, carboxyl, C1-C8 alkoxycarbonyl, toluenesulfonyl, toluenesulfonate, amino, mono-C1-C8 alkylamino, di-C1-C8 alkylamino, NHNH2, N3, haloacetamide, maleimide, succinimide, SH, SO3H, C1-C8 alkylsulfonyl, C1-C8 alkoxysulfonyl, 2-pyridyl disulfide, PO3H2, OPO3H2, -N≡C, -NCS, C4-C 10 Dienyl, C2-C8 alkenyl, C2-C8 alkynyl, C4-C 10 cycloalkynyl and C2-C8 alkynylcarbonyl, and R f It is H or C1-C8 alkyl.

13. The linker compound 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 linker compound according to claim 1, wherein L 1 C1-C 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 linker compound 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 is H or C1-C8 alkyl, and na and ma to mc are each independently an integer of 0 to 10.

16. The linker compound according to claim 1, wherein AL 1 -Selected from the group consisting of: R a1 C≡C-(CH2) n1 -; N3-(CH2) n2 -; R a1 , R b1 , R f1 , R f2 , R h1 , R h2 , R i1 and R d1 To R d10 are each independently H or C1-C8 alkyl, and R c1 , R e1 , R g1 , R h1 and R h2 Each is independently H, C1-C8 alkyl, C1-C8 alkylcarbonyl, C1-C8 alkoxycarbonyl or C6-C 12 Aryl-C1-C4 alkoxycarbonyl, n1 to n18 and m1 to m14 are each independently an integer of 0 to 10.

17. The linker compound according to claim 1, wherein The compound represented by Formula 1 is selected from the group consisting of compounds represented by the following formulae: Among them, A and L 1 , k, R 3 and PL is as defined in claim 1, and V" represents an electron withdrawing group.

18. The linker compound according to claim 1, wherein Formula A is represented by the following formula A-1: [Formula A-1] Where Z 1 , Z 2 , Z 3 , V, Y, T, W, PL, h, y and p are as defined in claim 1.

19. The linker compound 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, 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 is H or C1-C8 alkyl; 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.

20. The linker compound according to claim 19, wherein In the above formula 1-1 Select from the following structures: wherein q2 and q3 are as defined in claim 19.

21. The linker compound according to claim 19, 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 are each independently H or C1-C8 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.

22. The linker compound according to claim 19, wherein The compound represented by formula 1-1 is selected from the group consisting of compounds represented by the following formulae: Among them, q1 to q4, U 1 and U 2 As defined in claim 19, R d1 , R d5 and R e are each independently H or C1-C8 alkyl, and n1, n3, n4, n8, n14 and n15, m2, m8 and m10 are each independently an integer of 1 to 8.

23. A linker compound represented by the following formula B, or a pharmaceutically acceptable salt thereof: [Formula B] in, V' is -CH2-(L 2 ) l -PL; L 2 is a self-eliminating linker, which is selected so that the -CH2- of V' is 2 The cleavage of the bond between L 2 Cleavage of the bond with PL; PL is a heteroatom selected from N, O and S and L of V' 2 or -CH2-linked active agents; Z a and Z b One selected from N, NR 3 , O, S and Se, Z a and Z b The other of is CH or N; R 3 is H or a C1-C8 hydrocarbon 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 i, l, x and y are each independently 0 or 1, and p is an integer from 0 to 2.

24. The linker compound according to claim 23, wherein The compound is selected from the group consisting of compounds represented by the following formula:

25. A ligand-drug conjugate represented by the following formula 2, or a pharmaceutically acceptable salt thereof: [Formula 2] in, 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.

26. The ligand-drug conjugate according to claim 25, 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 C1-C 20 A group consisting of hydrocarbon groups, oligopeptides, polypeptides, fragments of antigenic polypeptides and artificial antibodies (Repebodies).

27. The ligand-drug conjugate according to claim 26, 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 an antibody, and modified immunoglobulin molecules comprising an antigen recognition site.

28. The ligand-drug conjugate according to claim 26, 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.

29. The ligand-drug conjugate according to claim 25, wherein E is an antibody, and the E-A' binding structure in Formula 2 includes Wherein * is the remaining part of the antibody.

30. The ligand-drug conjugate according to claim 25, wherein The conjugate represented by Formula 2 is selected from the following conjugates: Wherein mAb represents the antibody portion; V” is an electron-withdrawing group; m8 and n8 are each independently an integer from 1 to 10; Z 1 Selected from NR 3 , O, S and Se heteroatoms, R 3 is H or a C1-C8 hydrocarbon group; PL is part of the active agent; and n is a real number from 1 to 10.

31. The ligand-drug conjugate according to claim 30, wherein The ligand-drug conjugate represented by Formula 2 is selected from the group consisting of conjugates represented by the following formulae: where mAb is a portion of an antibody, and n is a real number from 1 to 10.

32. The ligand-drug conjugate according to claim 25, wherein The compound represented by Formula 2 is a compound represented by the following Formula 2-1: [Formula 2-1] in, 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 Each independently selected from And R e is H or C1-C8 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; n is a real number from 1 to 10.

33. The ligand-drug conjugate according to claim 32, wherein The conjugate represented by formula 2-1 is selected from the group consisting of conjugates represented by the following formulae: Wherein mAb is the antibody portion; q1 to q4, n, U 1 and U 2 As defined in claim 32; R d1 , R d5 and R e are each independently H or C1-C8 alkyl; and n1, n3, n4, n8, n14 and n15, m2, m8 and m10 are each independently an integer of 1 to 8.

34. The ligand-drug conjugate according to claim 32, wherein The conjugate represented by formula 2-1 is selected from the group consisting of conjugates represented by the following formulae: where mAb is a portion of an antibody, and n is a real number from 1 to 10.

35. A pharmaceutical composition comprising the ligand-drug conjugate according to any one of claims 25 to 34, and a pharmaceutically acceptable carrier or excipient.

36. The pharmaceutical composition according to claim 35, wherein The pharmaceutical composition is used for treating or preventing proliferative diseases, autoimmune diseases or infectious diseases.

37. An imaging composition comprising the ligand-drug conjugate according to any one of claims 25 to 34.

38. A composition for detection comprising the ligand-drug conjugate according to any one of claims 25 to 34.

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