Bioactive molecular conjugates, their preparation methods and uses

Abstract

This invention provides bioactive molecular conjugates, methods for preparing them, and their uses. [Solution] This disclosure provides novel bioactive molecular conjugates obtained by improving the coupling of the drug and targeting moiety in antibody-drug conjugates (ADCs) or small molecule drug conjugates (SMDCs), as well as methods for preparing the same and their use in the manufacture of pharmaceuticals for treating diseases associated with abnormal cell activity.

Description

Field of Invention

[0001] This disclosure relates to the field of medical technology, and concerns bioactive molecular conjugates, methods for preparing them, and, but not limited to, use in the prevention and / or treatment of neoplastic diseases, including abnormal Regarding the use of cellular activity and related diseases in the prevention and / or treatment of such diseases. [Background technology]

[0002] Chemotherapy was once the standard treatment for cancer, but it has a high biological activity that kills bacteria Sex molecules can mistakenly kill normal cells, leading to serious side effects. Targeted therapy can target Due to its sexual properties and antitumor activity, it has become a noteworthy research topic in the field of oncology. 2 Since the 20th century, biomolecular drugs (e.g., therapeutic antibodies or antibody fragments) and targeted small molecule ligans have been developed. Significant progress has been made in the development of antitumor drugs and tumor-targeted therapies using [unclear / unclear]. However, However, despite their high targeting potential, biomolecular drugs have limited effectiveness against solid tumors. It only has a mild healing effect; in addition, bioactive molecules often have no bioactive effect on cancer cells. Despite its high effectiveness in killing sex molecules, it lacks targeting ability and unintentionally damages normal cells, causing serious harm. It causes toxicity and side effects.

[0003] Recent research has shown that therapeutic antibodies can be linked to bioactive molecules to form antibody-drug conjugates. It has been found that ADCs can form antibodies (ADCs). ADCs have the target effect of antibodies and biological activity components. Because the activity of this molecule is combined, the bioactive molecule is called a "biological missile (biolog It is called an "ical missile." The ADC is guided by the antibody and targets It binds to cells, is then internalized by the cells, and releases the drug, thereby causing related diseases. The antibody's application value depends on its specificity and targeting ability to tumor cell-related targets. Not only is this reflected in the drug, but it also serves as an ideal carrier for targeted drug delivery, and the side effects of the drug are reduced. It reduces. Small molecule drug conjugates (SMDCs) are antibody-drug conjugates (A The same principle as in DC, namely, selection of receptors on the surface of tumor cells by a chemical process. It binds precisely, thereby targeting effector molecules (bioactive molecules) to tumor cells. Designed based on the coupling of bioactive molecules with several small molecule ligands that can be improved. The difference between SMDC and ADC is that SMDC uses small molecule ligands instead of antibodies. Furthermore, there are currently no SMDCs available on the market.

[0004] Currently, there are four commercially available ADCs: Mylotarg (Gemtsuz) Mab-ozogamicin), Adcetris (brentuximab-vedo) Chin, CD30 monoclonal antibody (MMAE), Kadcyla (tiger) Stuzumab-emtansine and Besponsa (Inotuzumab-ozo Gamycin, CD22 monoclonal antibody (calicheamicin). ADCs are generally antibodies It consists of a bioactive molecule and a linker. The bioactive molecule is covalently bonded to the antibody via the linker. They are coupled to each other; the antibody (e.g., monoclonal antibody) is applied to the surface of tumor cells. Because it can specifically recognize specific targets on the surface, ADCs can reach the surface of cancer cells. It guides ADCs to reach their destination and enter cancer cells through intracellular uptake. This enables the release of bioactive molecules in cancer cells, damaging normal tissue cells. It achieves the effect of specifically killing cancer cells without any other means.

[0005] Lysine is the most common binding site in antibodies, and the ε-amino group of lysine is phosphorus. It can react with the activated carboxyl group of Kerr to form an amide bond. Site-specific coupling Techniques for this are currently available, namely, the activation of the linker's carboxyl group. Then, it forms an amide bond with the specific lysine ε-amino group in the antibody, and the cup The ring is completed. However, such amide bonds are not affected by the action of enzymes in vivo. Under these conditions, they are easily hydrolyzed, and as a result, bioactive molecules and antibodies are dissolved before they can reach target cells. Release leads to increased toxicity, while simultaneously losing the targeting ability of ADCs.

[0006] The thio group of antibody cysteine ​​usually exists in the form of a disulfide bond. When the phido bond is opened, multiple free sulfhydryl groups can be obtained as coupling sites. Yes, it is possible. One method of coupling the free sulfhydryl group of the antibody is to... Michael addition reaction between hydryl groups and maleimide, or the formation of sulfur crosslinking bonds with a unique structure. This is achieved through two Michael addition reactions between a specific substrate and the free sulfhydryl group of the antibody. Yes, it exists. However, many studies describe AD obtained by the thiol-Michael addition method. It has been reported that C undergoes reverse Michael addition in the systemic circulation, which may lead to toxic reactions. International Publication No. 2016142049 describes amatoxy as a bioactive molecule. Bioactive compounds having the structure of methylsulfonyl-substituted benzobisoxadiazole The structure including the parent and linker is disclosed, but the details of coupling with the antibody are not specifically provided. Not described. (Details of the invention)

[0007] This invention relates to a coupling method for drug and targeting portion in ADC or SMDC. We disclose a novel bioactive molecule conjugate obtained by improving the aforementioned The denjugate offers high stability, extremely high coupling efficiency (90%), and high DAR. (5-8) is included. This disclosure is based on the above findings. Through vigorous research, the AD of the present invention has been found. C, for example, BT001021 (Example 32), showed biological activity in tumors after intravenous administration. Exposure to small molecule toxins was significantly higher in plasma, while Immu-132 was present at the same dose. Surprisingly, it was found that the plasma exposure was significantly higher than the tumor exposure under the pathway. Therefore, the ADC of this invention has a better therapeutic window than Immu-132. The researchers also found that the ADC of the present invention is effective in animal models of gastric cancer, breast cancer, and non-small cell lung cancer. I was surprised to find that it had higher efficacy than Immu-132.

[0008] A first aspect of this disclosure relates to a compound represented by formula (I) below or a pharmaceutically acceptable salt thereof. To provide. T-[L1-(L2)] m1 -(L3) m2 -(L4) m3 -E]-G Formula (I) [In the formula, T is a fragment of a bioactive molecule, preferably a fragment of a molecule having antitumor bioactivity; L1 is an amino acid, a peptide composed of 2 to 10 amino acids, an oligosaccharide, -(C H2) t1 -,-(CH2CH2O) t1 -(CH2) t2 -, [ka] Selected from; Here, each of R, R', R1 and R2 is independently H (hydrogen), deuteriu m), halogen, carboxylic acid group, sulfonic acid group, cyano, C 1~6 alkyl, halogenated C 1~6 alkyl (e.g., -CF3), cyano-substituted C 1~6 alkyl (for exa mple, -CH2CN), C 1~6 alkoxy, C 2~10 alkenyl, C 2~10 alkyn yl, C 3~6 cycloalkyl, 6- to 10-membered aryl or 5- to 12-membered heteroaryl, and each Z1 is independently an amino acid or a peptide composed of 2 to 10 amino acids, each of t1 and t2 is independently 0, 1, 2, 3, 4, 5 or 6, and each of x1 and x2 is independently 0, 1, 2, 3, 4, 5 or 6, each x3 is independently 0, 1, 2, 3 or 4, and L1 is bonded to T at position 1 of L1; L2 is selected from an amino acid, a peptide composed of 2 to 10 amino acids, an oligosaccharide, -(C H2) t1 -, -(CH2CH2O) t1 -(CH2) t2 -,

Chemical Formula

[0009] In some preferred embodiments, L1 is Val, Cit, Phe, Lys, D-Val Leu, Gly, Ala, Asn, peptides composed of 2-5 amino acids, [ka] Selected from; each of R, R', R1 and R2 is independently H (hydrogen), D (hydrogen) Rium), C 1~6 Alkyl, C 2~10 Alkenil, C 2~10 Alkinyl or C 3~ It is a 6-cycloalkyl group, and Z1 is Val, Cit, Phe, Lys, D-Val, Le u, Gly, Ala, Asn, Val-Cit, Cit-Val, Cit-Ala, Va l-Ala, Lys-Val, Val-Lys(Ac), Phe-Lys, Phe-Ly s(Ac), D-Val-Leu-Lys, Gly-Gly-Arg or Ala-Ala -Asn, where x1 is 0, 1, 2, or 3, and x3 is 0, 1, 2, 3, or 4.

[0010] In some preferred embodiments, L1 is Val, Cit, Phe, Lys, D-Val , Leu, Gly, Ala, Asn, Cit-Val, Val-Ala, Lys-Val , Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-L eu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, [ka] Selected from; R, R', and R1 are independently H (hydrogen) and D (deuterium). ), C 1~6 Alkyl, C 2~10 Alkenil, C 2~10 Alkinyl or C 3~6 Shik It is a rhalkyl, and Z1 is Val, Cit, Phe, Lys, D-Val, Leu, G ly, Ala, Asn, Val-Cit, Cit-Val, Cit-Ala, Val-A la, Lys-Val, Val-Lys(Ac), Phe-Lys, Phe-Lys(A c), D-Val-Leu-Lys, Gly-Gly-Arg or Ala-Ala-As n is such that x1 and x3 are independently 0, 1, 2, or 3.

[0011] In some preferred embodiments, L1 is Lys, Cit, Cit-Val, Val-A la, Lys-Val, [ka] Selected from; R, R', and R1 are independently H (hydrogen) and D (deuterium). ) or C 1~4 It is alkyl, and Z1 is Cit, Lys, Cit-Val, Cit-A la, Val-Ala, or Lys-Val, and x1 and x3 are each independently 0. It is either 1 or 2.

[0012] In some preferred embodiments, L1 is Lys, Cit, Cit-Val, Val-A la, Lys-Val, [ka] Selected from.

[0013] In some preferred embodiments, L1 is [ka] Selected from.

[0014] In some preferred embodiments, L2 is Val, Cit, Phe, Lys, D-Val Leu, Gly, Ala, Asn, peptides composed of 2-5 amino acids, [ka] Selected from; R3, R4, R5 and R6 each contain H (hydrogen) and D (hydrogen). (Mu), halogen, carboxylic acid group, sulfonic acid group, CF3, CN, CH2CN, C 1~4 a Lu kill, C 1~4 Alkoxy, C 2~6 Alkenil, C 2~6 Alkinyl or C 3~6 C Independently selected from chloroalkyls, y1 and y2 are independently 0, 1, 2, 3, 4 , 5, 6, 7 or 8, and L2 is connected to L1 at position 1 of L2; m1 is 0, 1, 2, or 3.

[0015] In some preferred embodiments, L2 is Val, Cit, Phe, Lys, D-Val , Leu, Gly, Ala, Asn, Val-Cit, Cit-Val, Val-Ala , Lys-Val, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac) , D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, [ka] Selected from; R3, R4, R5 and R6 each contain H (hydrogen) and D (hydrogen). (Mu), halogen, carboxylic acid group, sulfonic acid group, CF3, CN, CH2CN, C 1~4 a Lu kill, C 1~4 Alkoxy, C 2~6 Alkenil, C 2~6 Alkinyl or C3~6 C Independently selected from chloroalkyls, y1 and y2 are independently 0, 1, 2, 3, 4 , 5, 6, 7 or 8, and L2 is connected to L1 at position 1 of L2; m1 is 0, 1, or 2.

[0016] In some preferred embodiments, L2 is [ka] Selected from; R3, R4, R5 and R6 each contain H (hydrogen) and D (hydrogen). Mu) or C 1~4 Independently selected from alkyl groups, y1 and y2 are independently 0 and 1, respectively. , 2, 3, 4, 5, 6, 7 or 8, and L2 is connected to L1 at position 1 of L2; m1 is 1.

[0017] In some preferred embodiments, L2 is [ka] Selected from.

[0018] In some preferred embodiments, L2 is [ka] Selected from.

[0019] In some preferred embodiments, L3 may be replaced by one or more R7s. The bases: amino, 3-8 membered cycloalkylene, 3-8 membered aliphatic heterocyclylene, 6-12 6-12 member cross-linked heterocyclylene, 6-12 member spiroheterocyclylene, 6-12 member condensed heterocyclylene Criylene, 6-10 membered arylene, 5-12 membered heteroarylene, or 3-8 membered cycloal Selected from kylene-W-; W is oxygen or NR8, and R7 is H (hydrogen), D (hydrogen). Uterium, halogen, =O, CF3, CN, CH2CN, carboxyl, sulfonic acid group, C 1~4 Alkyl, C 1~4 Alkoxy, C 2~6 Alkenyl or C 2~6 Alkini Independently selected from the following; preferably 3-8 membered aliphatic heterocyclylenes, 6-12 membered crosslinked Heterocyclylene, 6-12 member spiroheterocyclylene, or 6-12 member condensed heterocyclylene Len has one or more nitrogen atoms; preferably a 3-8 membered aliphatic heterocyclylene. 6-12 member cross-linked heterocyclylene, 6-12 member spiroheterocyclylene, or 6-12 member contracted heterocyclylene The heterocyclylene has one or more quaternary nitrogen atoms; preferably, a 3-8 membered resin. Alliphatic heterocyclylene, 6-12 membered cross-linked heterocyclylene, 6-12 membered spiroheterocycline Lylene or 6-12 membered condensed heterocyclylene has one or more nitrogen atoms, and at least One nitrogen atom is substituted with =O; R8 is H (hydrogen), D (deuterium) , C 1~6 Alkyl, C 2~6 Alkenil, C 3~6 Alkinyl, C 3~6 Cycloalkyl Ru, C 1~6 Alkoxy or cyanoC 1~2 Selected independently from alkyl; m2 is 0, 1, 2, or 3.

[0020] In some preferred embodiments, L3 may be replaced by one or more R7s. The base is selected from amino, 3-6 member aliphatic heterocyclylene, or 5-10 member heteroarylene. Selected; R7 is H (hydrogen), D (deuterium), halogen, =O, CF3, CN, CH2CN, carboxyl, sulfonic acid group, C 1~4 Alkyl, C 1~4 Alkoxy, C 2~6 Alkenyl or C 2~6 Independently selected from alkynyls; preferably, 3-6 membered lipids. Aliphatic heterocyclylenes have one or more nitrogen atoms; preferably, 3-6 membered aliphatic heterocyclylenes. Telocyclylene has one or more quaternary nitrogen atoms; preferably 3-6 membered aliphatic atoms. Heterocyclylene has one or more nitrogen atoms, and at least one nitrogen atom is =O. It has been replaced; m2 is 0, 1, or 2.

[0021] In some preferred embodiments, L3 may be replaced by one or more R7s. The base is selected from amino or 5-6 member heteroarylenes; R7 is H (hydrogen), D (di Deuterium, halogen, =O, CF3, CN, CH2CN, carboxyl, sulfone acid group, C 1~4 Alkyl, C 1~4 Alkoxy, C 2~6 Alkenyl or C 2~6 Alki Selected independently of nil; m2 is either 0 or 1.

[0022] In some preferred embodiments, L3 may be replaced by one or more R7s. The base is selected from amino, N-methylpiperidylene, pyrazolylene, or triazolylene. R7 is H (hydrogen), D (deuterium), halogen, =O, CF3, CN, CH2 CN, carboxyl, sulfonic acid group, C 1~4 Alkyl, C 1~4 Alkoxy, C 2~6 Alkenyl or C 2~6 Selected independently from alkinyl; m2 is 0 or 1.

[0023] In some preferred embodiments, L3 is selected from triazolylene; m2 is 0 or 1 .

[0024] In some preferred embodiments, L3 is

Chemical Formula

[0025] In some preferred embodiments, L3 is the following group optionally substituted with one or more R7 : amino,

Chemical Formula

[0026] In some preferred embodiments, L3 is

Chemical Formula

[0027] In some preferred embodiments, L3 is [ka] Selected from.

[0028] In some preferred embodiments, L4 is [ka] Selected from, Z4 is a 6-10 member aryl or a 5-6 member heteroaryl; R 10 teeth H (hydrogen) or C 1~6 Selected from alkyl; Z2 is C 1~6 Alkylene, C 2~1 0 Alkenylene, C 2~10 Alkinirene and C 3~8 Selected from cycloalkylenes; R9 is either H (hydrogen) or C 1~6 Selected from alkyl groups; Z3 is absent or C 1~6 Selected from alkylene; or R9 and Z3 together with the nitrogen atom bonded to them, 4 It forms an 8-membered heterocyclene; α is independently 0, 1, 2, 3, 4, 5, or 6. L4 is bonded to E at position 2; m3 is 0, 1, 2, or 3.

[0029] In some preferred embodiments, L4 is [ka] Selected from, Z4 is a benzene ring, and R 10 H (hydrogen) and C 1~6 From alkyl Selected; Z2 is C 1~6 Alkylene, C 2~10 Alkenylene, C2~10 Alkini Len or C 3~8 Selected from cycloalkylenes; R9 is H (hydrogen) or C 1~6 Alki Selected from the list; Z3 does not exist or C 1~6 Selected from alkylenes, or R 9 and Z3, together with the nitrogen atom bonded to them, form a 4- to 8-membered heterocyclene. α is independently 0, 1, 2, 3, 4, 5, or 6, and L4 is bonded to E at position 2 of L4. L4 is bonded to E at position 2; m3 is 0, 1, 2, or 3.

[0030] In some preferred embodiments, L4 is [ka] Selected from, Z4 is a 5-6 member heteroarylene; R 10 is H (hydrogen) or C 1~ Selected from 6 alkyl groups; Z2 is C 1~6 Alkylene, C 2~10 Alkenylene, C2 ~10 Alkinirene and C 3~8 Selected from cycloalkylenes; R9 is H (hydrogen) or C 1~6 Selected from alkyl groups; Z3 is absent or C 1~6 Selected from alkylene R9 and Z3, together with the nitrogen atom bonded to them, form a 4-8 member heterozygous molecule. It forms rylene; α is independently 0, 1, 2, 3, 4, 5 or 6; L4 is L4 It is coupled to E in second place; m3 is 0, 1, 2, or 3.

[0031] In some preferred embodiments, L4 is [ka] Selected from; m3 is 1.

[0032] In some preferred embodiments, L4 is [ka] Selected from; m3 is 1.

[0033] In some preferred embodiments, L4 [ka] Selected from; m3 is 1.

[0034] In some preferred embodiments, E is one or more R 12 5 may be replaced with Selected from ~10-membered heteroarylenes; R 12 H (hydrogen), D (deuterium) , halogen, CN, nitro, C 1~4 Alkyl or halogenated C 1~4 Independent from alkyl Selected.

[0035] In some preferred embodiments, E is one or more R 12 The next one may be replaced by The base is selected from pyrimidilene, quinolylene, or pyrrolo[2,3-d]pyrimidilene; R 12 H (hydrogen), D (deuterium), halogen, CN, nitro, C 1~2 Al Kill or halogenated C 1~2 It is selected independently of alkyl.

[0036] In some preferred embodiments, E is one or more R 12 P may be replaced with Selected from limidinil; R 12 is independently selected from H (hydrogen) or D (deuterium). It will be done.

[0037] In some preferred embodiments, G is a halogen, OMs, OTs, OTf, nitro, or is one or more R 13 The following groups may be substituted: alkylthio, arylthio O, heteroarylthio, alkylsulfinyl, arylsulfinyl, heteroaryl Lusulfinyl, alkylsulfonyl, arylsulfonyl, or heteroarylsulfonyl One of the Honils is selected; R 13 H (hydrogen), D (deuterium), H Logen, CN, Nitro, C 1~6 Alkyl, halogenated C 1~6 Alkyl, C 1~6 Al The coxy, 6-10 membered aryl, or 5-12 membered heteroaryl is independently selected.

[0038] In some preferred embodiments, G is F, Cl, Br, I, OMs, OTs, OTf, Methylsulfonyl, ethylsulfonyl, p-toluenesulfonyl, or naphthalenesulfonyl Selected from the options.

[0039] In some preferred embodiments, G is F, Cl, Br, OMs, OTs, methyl sulfon Selected from nyl or p-toluenesulfonyl.

[0040] In some preferred embodiments, G is selected from Cl or methylsulfonyl.

[0041] In some preferred embodiments, [ka] In this, G is preferably methylsulfonyl and E is preferably pyrimidylene. m3 is 1.

[0042] In some preferred embodiments, [ka] teeth [ka] Here, m4 is preferably an integer from 0 to 6, and methylsulfonyl is in the pyrimidine ring. It is a substituent on a carbon atom adjacent to a nitrogen atom.

[0043] In some preferred embodiments, [ka] teeth [ka] Here, m5 is preferably an integer from 0 to 6, and methylsulfonyl is in the pyrimidine ring. It is a substituent on a carbon atom adjacent to a nitrogen atom.

[0044] In some preferred embodiments, [ka] teeth [ka] Here, m6 is preferably an integer from 0 to 6, and methylsulfonyl is in the pyrimidine ring. It is a substituent on a carbon atom adjacent to a nitrogen atom.

[0045] In some preferred embodiments, [[Chem.]] is [[Chem.]] , m7 is an integer of 1 to 5, and methylsulfonyl is attached to a nitrogen atom in the pyrimidine ring as a substituent on an adjacent carbon atom.

[0046] In some preferred embodiments, [[Chem.]] is [[Chem.]] , m8 is an integer of 1 to 5, and methylsulfonyl is attached to a nitrogen atom in the pyrimidine ring as a substituent on an adjacent carbon atom.

[0047] In some preferred embodiments, [[Chem.]] is [[Chem.]] , m9 is an integer of 1 to 5, and R 13 is selected from hydrogen or C 1~6 alkyl, methylsulfonyl is a substituent on a carbon atom adjacent to a nitrogen atom in the pyrimidine ring.

[0048] In some preferred embodiments, [[Chem.]] is [[Chem.]] , and m 10 is an integer of 0 to 6, and Z4 is selected from 5- to 6-membered heteroarylene; methylsulfonyl is a substituent on a carbon atom adjacent to a nitrogen atom in the pyrimidine ring.

[0049] In some preferred embodiments,

Chemical Structure

Chemical Structure

[0050] In some preferred embodiments,

Chemical Structure

Chemical Structure

[0051] In some preferred embodiments,

Chemical Structure

Chemical Structure

[0052] In some preferred embodiments, [ka] teeth [ka] And, m 10 Z4 is an integer from 0 to 6, and Z4 is selected from 6 to 10 members of arylene; Methyl The sulfonyl substituent is a substituent on a carbon atom adjacent to a nitrogen atom. More preferably, m 10 is an integer between 0 and 6.

[0053] In some preferred embodiments, [ka] teeth [ka] And, m 10 Z is an integer between 0 and 6, and Z4 is a benzene ring.

[0054] In some preferred embodiments, [ka] teeth [ka] That is the case.

[0055] In some preferred embodiments, the formula (I) [ka] The following fragments are selected. [ka] [ka] [ka] [ka]

[0056] In some preferred embodiments, T is a fragment of a bioactive molecule. In this state, the bioactive molecule is a platinum metal complex (e.g., oxaliplatin) or a gold metal complex, etc. Metal complexes; such as bleomycin or pingyangmycin. Glycopeptide antibiotics; topoisomerase I inhibitors (e.g., camptothecin, hidol) Roxycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, topote Can, bellotencian, or rubitecan, or topoisomera -ase II inhibitors (e.g., actinomycin D, doxorubicin, duocalmycin, DNA (such as daunorubicin, mitoxantrone, podophyllotoxin, or etoposide) Topoisomerase inhibitors; methotrexate, 5-fluorouracil, cytarabine, gem Cytabine, mercaptopurine, pentostatin, fludarabine, cladribine or narabi Drugs that interfere with DNA synthesis, such as narabine; tubulin inhibitors, pymebrane Rustin alkaloids, vincristine, vinblastine, paclitaxel, docetaxel Drugs that act on structural proteins such as iodine or cabazitaxel; serine / threonine kinase Histidine kinase inhibitors, tyrosine kinase inhibitors, aspart kinase inhibitors, or histidine kinase inhibitors Inhibitors such as tumor cell signaling pathway inhibitors; proteasome inhibitors; histone desaturates Histone deaceylase inhibitors; tumor angiogenesis inhibitors; cysts Clin inhibitors; meitansine derivatives; calicheamycin derivatives; auristatin derivatives; Pyrrolobenzodiazepine dimer (PBD) derivatives; melphalan; mitomycin C; Lorambucil; and inhibitors of tumor cell proliferation, and promotes tumor cell apoptosis or necrosis. It is selected from other active substances.

[0057] In some preferred embodiments, the bioactive molecule is [ka] [In the formula, R 14 R 15 Selected from acyl or sulfonyl substituted with R 15 C 1~6 Alkyl, halogenated C 1~6 Alkyl, 6-10 membered aryl, or 5-12 membered aryl Selected from member heteroaryls; R 16 These are H (hydrogen), D (deuterium), and C 1~ 6 alkyl, or R 17 C is replaced by 1~6 Selected from alkyl, R 17 This These are not limited to, but include selected aryl or heteroaryls containing phenyl and pyridyl. Selected, m 11 [This value is 0, 1, or 2.] Selected from.

[0058] In some preferred embodiments, the bioactive molecule is [ka] [In the formula, R 14 R 15 Selected from acyl or sulfonyl substituted with R 15 C 1~6 Alkyl, halogenated C 1~6 Alkyl, 6-10 membered aryl, or 5-12 membered aryl Selected from member heteroaryls; R 16 These are H (hydrogen), D (deuterium), and C 1~ 6 alkyl, R 17 C is replaced by 1~6 Selected from alkyl, R 17 These Not limited to, but selected from aryl or heteroaryl phenyl or pyridyl. re, m 11 [This value is 0, 1, or 2.] Selected from.

[0059] In some preferred embodiments, the bioactive molecule is [ka] Selected from.

[0060] In some preferred embodiments, the bioactive molecule is [ka] Selected from.

[0061] In some preferred embodiments, the bioactive molecule is [ka] Selected from.

[0062] In some preferred embodiments, the bioactive molecule is [ka] Selected from.

[0063] In some preferred embodiments, the bioactive molecule [ka] Selected from.

[0064] In some preferred embodiments, T is [ka] [ka] [ka] Selected from.

[0065] In some preferred embodiments, T is [ka] [ka] [ka] Selected from.

[0066] In some preferred embodiments, T is [ka] Selected from.

[0067] In some preferred embodiments, T is [ka] Selected from.

[0068] In some preferred embodiments, T is [ka] Selected from.

[0069] In some preferred embodiments, the compound represented by formula (I) is [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] Selected from.

[0070] In some preferred embodiments, the compound is [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] Selected from.

[0071] In a second aspect, the disclosure includes a bioactive molecule, a linker and a targeting portion. It provides a denjugate. The targeting portion is activated via an active group (e.g., a thiol group). It is connected to a linker to form a conjugate.

[0072] In some preferred embodiments, the structure of the conjugate is the structure shown in formula (II) below. That is the case. {T-[L1-(L2) m1 -(L3) m2 -(L4) m3 -E]} γ -A Equation (I I) [In the formula, A is the targeting portion (e.g., small molecule ligand, protein, polypeptide or non- It is a protein reagent (e.g., sugar, RNA, or DNA); γ is an integer or small integer between 1 and 10. It is a number; preferably, γ is an integer or decimal between 5 and 8 (for example, 5, 6, 7, or 8). ; The remaining components are as described in the first aspect of this disclosure.

[0073] In some preferred embodiments, the target of A is epidermal growth factor, Trop-2, CD37, HER2, CD70, EGFRvIII, Mesothelin, Folic Acid Folate receptor 1, Mucin 1, CD138, C D20, CD19, CD30, SLTRK6, Nectin4, tissue factor ( Tissue factor), mucin 16, endothelin receptor (En dothelin receptor), STEAP1, SLC39A6, guanylate receptor), STEAP1, SLC39A6, Clase (Guanylylcyclase) C, PSMA, CCD79b, CD22, Sodium phosphate cotransporter ter)2B, GPNMB, Trophoblast glycoprotein (Trophoblast g lycoprotein), AGS-16, EGFR, CD33, CD66e, CD74 CD56, PD-L1, TACSTD2, DR5, E16, 0772P, MPF, Na pi3b, Sema 5b, PSCA hlg, ETBR, MSG783, STEAP2 TrpM4, CRIPTO, CD21, CD79b, FcRH2, NCA, MDP, I L20Rα レカン(Brevican), EphB2R, ASLG659, PSC A, GEDA, BAFF-R, CD79a, CXCR5, HLA-DOB, P2X5 D72、LY64、FcRH1、IRTA2、TENB2、イテグリン(integr in) α5β6, α4β7, FGF2, FGFR2, Her3, CA6, DLL3, DL L4, P-cadherin, EpCAM, pCAD, CD223, LY PD3, LY6E, EFNA4, ROR1, SLITRK6, 5T4, ENPP3, クロ Claudin (Claudin)18.2, BMPR1B, Tyro7, c-Met, Apo E, CD1 lc, CD40, CD45(PTPRC), CD49D(ITGA4), C D80, CSF1R, CTSD, GZMB, Ly86, MS4A7, PIK3AP1, P.S IK3CD, CCR5, IFNG, IL10RA1, IL-6, ACTA2, COL7A 1. LOX, LRRC15, MCPT8, MMP10, NOG, SERPINEl, ST AT1, TGFBR1, CTSS, PGF, VEGFA, C1QA, C1QB, ANGP TL4, EGLN, EGLN3, BNIP3, AIF1, CCL5, CXCL10, CX CL11, IFI6, PLOD2, KISS1R, STC2, DDIT4, PFKFB3 PGK1, PDK1, AKR1C1, AKR1C2, CADM1, CDH11, COL 6A3, CTGF, HMOX1, KRT33A, LUM, WNT5A, IGFBP3, M MP14, CDCP1, PDGFRA, TCF4, TGF, TGFB1, TGFB2, C D1 lb, ADGRE1, EMR2, TNFRSF21, UPK1B, TNFSF9, Choose from MMP16, MFI2, IGF-1R, RNF43, NaPi2b, or BCMA. It will be done.

[0074] In some preferred embodiments, A is a folic acid derivative, a glutamate urea derivative, or somatos Tatin derivatives, aryl sulfonamide derivatives (e.g., carbonic anhydrase IX inhibitors), two Polyenes, cyanine dyes, or IR-783 or their derivatives that link aliphatic indoles. It is a small molecule ligand, such as a conductor.

[0075] In some preferred embodiments, A is [ka] Selected from.

[0076] In some preferred embodiments, A is an antibody such as a monoclonal antibody or its antigen binding. A fragment, where the monoclonal antibody or its antigen-binding fragment is Fab, Fab', F(ab')2, Fd, Fv, dAb, complementation-determining fragment, single-chain antibody (e.g., scFv) Non-human antibodies, humanized antibodies, chimeric antibodies, fully humanized antibodies, probodies, bispecific antibodies or includes multispecific antibodies.

[0077] In some preferred embodiments, A is an anti-Her2 bloc such as trastuzumab or pertuzumab. Monoclonal antibodies; or anti-Trop-2 monoclonal antibodies such as sacituzumab. .

[0078] In some preferred embodiments, A is an anti-Trop-2 antibody such as M1, M2, or M3. It is a noclonal antibody. [Table 1]

[0079] The designation of amino acids in each region or domain is by Chothia & Lesk (19 87) J.Mol.Biol.196:901~917;Chothia et al. (1989) This can be according to the definition in Nature 342:878-883. 1. Heavy and light chain sequences of the hydrophobic modified antibody M1 Amino acid sequence of the heavy chain variable region of M1: (121aa) QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVK QAPGQGLKWMGWINTDSGEPTYTDDFKGRFAFSLDTSVST AYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVT VSS (Sequence ID 11) Amino acid sequence of the light chain variable region of M1: (107aa) DIQLTQSPSSLSASVGDRVSITCKASQDVSSAVAWYQQ KPGKAPKLLIYSASYRYTGVPDRFSGSGSGTDFTLTISSL QPEDFAVYYCQQHYSTPLTFGAGTKVEIK(Sequence ID 12) 2. Heavy and light chain sequences of the hydrophobic modified antibody M2 Amino acid sequence of the heavy chain variable region of M2: (121aa) QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVK QAPGQGLKWMGWINTDSGEPTYTDDFKGRFAFSLDTSVST AYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVT VSS (Sequence ID 13) Amino acid sequence of the M2 light chain variable region: (107aa) DIQLTQSPSSLSASVGDRVSITCKASQDVSSAVAWYQQ KPGKAPKLLIYSASYRYTGVPDRFSGSGSGTDFTLTISSL QPEDFAVYYCQQHYITPLTFGAGTKVEIK(Sequence ID 14) 3. Heavy and light chain sequences of hydrophobic modified antibody M3 Amino acid sequence of the heavy chain variable region of M3: (121aa) QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVK QAPGQGLKWMGWINTDSGEPTYTDDFKGRFAFSLDTSVST AYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVT VSS (Sequence ID 15) Amino acid sequence of the light chain variable region of M3: (107aa) DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQ KPGKAPKLLIYSASYRYTGVPDRFSGSGSGTDFTLTISSL QPEDFAVYYCQQHYSTPLTFGAGTKVEIK (Sequence ID 16) Sequence of the light chain constant regions M1, M2, and M3: (107aa) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC (Sequence ID 9) Sequence of the heavy chain constant regions of M1, M2, and M3: (330aa) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK (Sequence ID 10)

[0080] The terminal lysine molecules of the heavy chain are easily deleted, but such deletions do not affect biological activity. . Dick, LW et al., Biotechnol.Bioeng., 100:1132~ See 1143. The above monoclonal antibody M1, in which Lys is deleted at the end of the heavy chain, M2, M3 and their sequences or fragments are all M1, M2, M3 monoclonas of the present invention. It corresponds to a ru antibody.

[0081] In some preferred embodiments, A recognizes the cell surface integrin receptor RGD. Petit; a compound that recognizes cell surface growth factor receptors such as EGF, PDGF, or VEGF. Growth factors; or functional cell surface plasminogen activator, bombesin, bradykinin, Selected from somatostatin or peptides capable of recognizing prostate-specific membrane antigen receptors. .

[0082] In some preferred embodiments, A is CD40 ligand, CD30 ligand, OX40 Ligands, PD-1 ligand, ErbB ligand, Her2 ligand, TACSTD2 ligand Selected from either Gand or DR5 ligand.

[0083] In some preferred embodiments, the conjugate is selected from the following: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [In the formula, γ is an integer or decimal between 1 and 10, and mAb is anti-Trop-2 monoclonal anti- It is a monoclonal antibody or an anti-Her2 monoclonal antibody; preferably, an anti-Trop-2 monoclonal antibody. The antibody is selected from sacituzumab, M1, M2, or M3 antibodies, and is an anti-Her2 monoclonal antibody. The ophthalmic antibody is trastuzumab or pertuzumab; preferably, γ is 5-8 (e.g.) It is an integer or decimal number of 5, 6, 7, or 8.

[0084] In some preferred embodiments, the conjugate is selected from the following: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [In the formula, γ is an integer or decimal between 1 and 10, and mAb is anti-Trop-2 monoclonal anti- It is a monoclonal antibody or an anti-Her2 monoclonal antibody; preferably, an anti-Trop-2 monoclonal antibody. The anti-Her2 monoclonal antibody was selected from sacituzumab, and the anti-Her2 monoclonal antibody was selected from trastuzumab or The γ is selected from pertuzumab; preferably, γ is 5-8 (e.g., 5, 6, 7, or 8) It is an integer or a decimal number.

[0085] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [ka] [ka] [In the formula, A1 is sacituzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between ~7.5, 6.5~8, 7~8, or 7.5~8.

[0086] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [In the formula, A1 is sacituzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between ~7.5, 6.5~8, 7~8, or 7.5~8.

[0087] In some preferred embodiments, the conjugate is as follows: [ka] [In the formula, A1 is sacituzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between ~7.5, 6.5~8, 7~8, or 7.5~8.

[0088] In some preferred embodiments, the conjugate is as follows: [ka] [In the formula, A1 is a fragment of sacituzumab, and γ is an integer or decimal between 1 and 10; preferred γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7. It is an integer or decimal between 6.5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8.

[0089] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [ka] [ka] [In the formula, A2 is trastuzumab, and γ is an integer or decimal number between 1 and 10; preferably] γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6. It is an integer or decimal between 5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8.

[0090] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [In the formula, A2 is trastuzumab, and γ is an integer or decimal number between 1 and 10; preferably] γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6. It is an integer or decimal between 5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8.

[0091] In some preferred embodiments, the conjugate is as follows: [ka] [In the formula, A2 is trastuzumab, and γ is an integer or decimal number between 1 and 10; preferably] γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6. It is an integer or decimal between 5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8.

[0092] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [ka] [ka] [In the formula, A3 is pertuzumab, and γ is an integer or decimal number between 1 and 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between ~7.5, 6.5~8, 7~8, or 7.5~8.

[0093] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [In the formula, A3 is pertuzumab, and γ is an integer or decimal number between 1 and 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between ~7.5, 6.5~8, 7~8, or 7.5~8.

[0094] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [ka] [ka] [In the formula, A4 is antibody M1, and γ is an integer or decimal number between 1 and 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 0.5, 6.5-8, 7-8, or 7.5-8.

[0095] In some preferred embodiments, the conjugate is as follows: [ka] [In the formula, A4 is antibody M1, and γ is an integer or decimal number between 1 and 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 0.5, 6.5-8, 7-8, or 7.5-8.

[0096] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [ka] [ka] [In the formula, A5 is antibody M2, and γ is an integer or decimal number between 1 and 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 0.5, 6.5-8, 7-8, or 7.5-8.

[0097] In some preferred embodiments, the conjugate is as follows: [ka] [In the formula, A5 is antibody M2, and γ is an integer or decimal number between 1 and 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 0.5, 6.5-8, 7-8, or 7.5-8.

[0098] In some preferred embodiments, the conjugate is as follows: [ka] [ka] [ka] [ka] [In the formula, A6 is antibody M3, and γ is an integer or decimal number between 1 and 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 0.5, 6.5-8, 7-8, or 7.5-8.

[0099] In some preferred embodiments, the conjugate is as follows: [ka] [In the formula, A6 is antibody M3, and γ is an integer or decimal number between 1 and 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 0.5, 6.5-8, 7-8, or 7.5-8.

[0100] In another aspect, the present disclosure relates to a method for preparing a conjugate of a second aspect. The step involves coupling the linker of the compound of formula (I) with the active group of the targeting moiety. Provides a method that includes the pu.

[0101] In some preferred embodiments, the method reduces the disulfide bond of the targeting portion. The process includes the step of opening the cells with an agent (e.g., TCEP) to obtain a sulfhydryl group.

[0102] In some preferred embodiments, the method involves linking the compound of formula (I) with a targeting The process includes the step of forming a CS bond between the portion and the sulfhydryl group.

[0103] In some preferred embodiments, the targeting portion is an anti-Her2 monoclonal antibody. (For example, trastuzumab, pertuzumab) or anti-Trop-2 monoclonal antibody (For example, sacituzumab, M1, M2, or M3), or their active fragments or variants ru.

[0104] In some preferred embodiments, the molar ratio of formula (I) of the targeting portion to the compound The ratio is 1:(1~20); preferably, the coupling is carried out in water and / or an organic solvent. Preferably, the organic solvent is N,N-dimethylformamide, dimethyl sulfoxide , N-methylpyrrolidone, nitrile (e.g., acetonitrile), alcohol (e.g., me Selected from ethanol, ethanol, or any combination thereof.

[0105] In some preferred embodiments, the method further extends the step of purifying the coupling product. The coupling product is preferably chromatographic (e.g., ion exchange chromatographic). Chromatography, hydrophobic chromatography, reverse-phase chromatography, or affinity chromatography It is purified by one or more methods of chromatography.

[0106] In another aspect, the Disclosure may relate to the compounds of the first aspect of the Disclosure or their pharmaceutically acceptable properties. A pharmaceutical composition comprising a salt or a conjugate of the second embodiment, and one or more pharmaceutical excipients. To provide goods.

[0107] In another aspect, the disclosure relates to treating diseases associated with abnormal cell activity (e.g., cancer). In the manufacture of pharmaceuticals, the compound of the first embodiment or a pharmaceutically acceptable salt thereof A second embodiment of the use of conjugates is provided.

[0108] In some preferred embodiments, the cancer is esophageal cancer (e.g., esophageal adenocarcinoma, esophageal squamous cell carcinoma). Squamous cell carcinoma), brain tumors, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), squamous cell carcinoma, Bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, Endometrial cancer, colorectal cancer, liver cancer, kidney cancer, non-Hodgkin lymphoma, central nervous system tumors Surgery (e.g., glioma, glioblastoma multiforme, glioma, or sarcoma), prostate cancer, and thyroid cancer These are solid tumors or non-solid tumors.

[0109] In another aspect, the disclosure relates to the treatment of diseases associated with abnormal cell activity (e.g., cancer). or a compound of the first embodiment or a pharmaceutically acceptable salt thereof or a conjugate of the second embodiment The use of a steroid or pharmaceutical composition is provided.

[0110] In another aspect, this disclosure relates to a method for treating diseases associated with abnormal cell activity (e.g., cancer). A law that specifies an effective amount of the compound in the first embodiment or a pharmaceutically acceptable salt thereof or the second A step of administering the conjugate or pharmaceutical composition of the form to an individual that requires it. This provides a method that includes this.

[0111] Unless otherwise specified, all scientific and technical terms used in this disclosure are intended for those skilled in the art. Therefore, it has a generally understood meaning. Furthermore, the cell culture and molecular genetic information used herein All experimental procedures in genetics, nucleic acid chemistry, and immunology are widely used in their respective fields. This is a simple step. In addition, the definitions and explanations of related terms are provided for a better understanding of this disclosure. For this purpose, see below.

[0112] In this disclosure, pharmaceutical excipients refer to excipients and additives used in pharmaceutical and formulation processes. It is a substance that has been evaluated as reasonable in terms of safety, and is included in pharmaceutical formulations in addition to the active ingredient. In addition to being used as excipients, carriers, and stability enhancers, pharmaceutical excipients are also used. Furthermore, it possesses important functions such as solubilization and sustained release, which affect the quality, safety, and efficacy of the drug. It is an important component that can have an effect. Pharmaceutical excipients are natural substances, semi-synthetic substances, depending on the source. They can be divided into crystalline and total synthetic substances; depending on their function and use, they are used as solvents, propellants, and solubilizers. Agents, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators Stabilizers, flow enhancers, flavoring agents, preservatives, suspending agents, coating substances, fragrances, anti-adhesion agents. Agents, antioxidants, chelating agents, permeability enhancers, pH adjusters, buffers, plasticizers, surfactants, Foaming agents, defoaming agents, thickeners, encapsulating agents, wetting agents, absorbents, diluents, flocculants and descaling agents, They can be divided into release adjuvants and release delayants; depending on the route of administration, they can be administered orally, by injection, or via smear. It can be classified into membrane, transdermal or topical administration, nasal or oral inhalation, and ocular administration. Additives can be used for pharmaceutical formulations with different routes of administration, resulting in different effects and uses. It holds.

[0113] The pharmaceutical composition is available in tablet, capsule, granule, oral solution, oral suspension, and via the route of administration. Oral emulsions, powders, tinctures, syrups, injections, suppositories, ointments, creams, pastes ophthalmic preparations, pills, subdermal preparations, aerosols, powders and sprays, etc. It can be formulated into various appropriate dosage forms. The pharmaceutical composition or appropriate dosage form is disclosed herein. The compound or its pharmaceutically acceptable salt or conjugate 0.01 mg to 1000 mg mg, appropriately 0.1 mg to 800 mg, preferably 0.5 to 500 mg, preferably 0 It may contain 0.5 to 350 mg, and particularly preferably 1 to 1 to 250 mg.

[0114] The pharmaceutical composition is administered in the form of an injectable preparation, including an injectable solution, a sterile powder for injection, and a concentrated solution for injection. It can be provided. Acceptable carriers and solvents include water, Ringer's solution, and isotonic sodium chloride. It contains a lye solution. In addition, it also contains sterile, non-volatile oils such as monoglycerides or diglycerides. It can be used as a solvent or suspension medium.

[0115] In this disclosure, the term “individual” includes human individuals or non-human animals. Human individuals include human individuals (referred to as patients) who have a disease (for example, a disease described herein). This includes individuals that are (or are normal). The term "non-human animal" in this disclosure refers to non-mammalian animals. non-human primates (e.g., birds, amphibians and reptiles), livestock, and / or domestic animals ( For example, all vertebrates, such as mammals (e.g., sheep, dogs, cats, cows, and pigs), are included. Born.

[0116] In this disclosure, the term “effective dose” means the amount of one or more symptoms of the treated disease after administration. This refers to the amount of compound that reduces the effect to a certain extent.

[0117] In this disclosure, the term “conjugate” means a bioactive molecule with a targeting moiety. This refers to a substance obtained by linking. In some embodiments of this disclosure, a bioactive molecule This is linked to the targeting portion via a linker. The linker is in a specific environment (for example, In an intracellular low pH environment, or under specific action (e.g., action of lysosomal proteases). This allows for cleavage, thereby dissociating the bioactive molecule from the targeting region. In some embodiments of this disclosure, the linker is a peptide or a disulfide bond, etc. Includes units that can be cleaved or cannot be cleaved. In some embodiments of this disclosure, a bioactive molecule is used. It is cleaved in a different environment or under different conditions, thereby separating the bioactive molecule from the targeting portion. It is directly linked to the targeting portion by a separable covalent bond.

[0118] In this disclosure, the terms "bioactive substance" and "bioactive molecule" refer to substances that inhibit or inhibit cellular function. This refers to a substance that inhibits and / or causes cell death or destruction. (Some embodiments of this disclosure) Therefore, the bioactive substance or bioactive molecule in the conjugate is a molecule that has antitumor bioactivity. For example: At211, I131, I125, Y90, Re186, Re188, Radioactive isotopes such as Sm153, Bi212, P32, Pb212, or radioactive isotopes of Lu Body; metal complexes such as platinum metal complexes, gold metal complexes, or oxaliplatin; bleomycin or These include glycopeptide antibiotics such as pinyanmycin; topoisomerase I inhibitors, for example. Camptothecin, Hydroxycamptothecin, 9-Aminocamptothecin, SN-38 Irinotecan, topothecan, berotensian, rubitecan, or topoisomer Ze II inhibitors, for example, actinomycin D, doxorubicin, duocalmycin, da DNA topois such as unorubicin, mitoxantrone, podophyllotoxin, and etoposide Somerase inhibitors; methotrexate, 5-fluorouracil, cytarabine, gemcitabine , mercaptopurine, pentostatin, fludarabine, cladribine or narabine, etc. Drugs that interfere with DNA synthesis; tubulin inhibitors, vinblastine alkaloids, vin Cristine, vinblastine, paclitaxel, docetaxel, or cabazitaxel, etc. Drugs that act on structural proteins; serine / threonine kinase inhibitors, tyrosine kinase inhibitors Tumor cell signals such as inhibitors, aspart kinase inhibitors, or histidine kinase inhibitors Inhibitors of signaling pathways; also, proteasome inhibitors; histone decayase inhibitors; tumor Angiogenesis inhibitors; cyclin inhibitors; meitansine derivatives; calicheamycin derivatives; o - Listatin derivatives; pyrrolobenzodiazepine (PBD) derivatives; melphalan; mito Mycin C; chlorambucil; and inhibitors of tumor cell proliferation, tumor cell apoptosis Other active substances that promote necrosis or other necrotic substances; enzymes such as nucleolytic enzymes and their fragments; antibiotics; anesthetics Small molecule toxins derived from bacteria, fungi, plants or animals, including fragments and / or variants thereof This includes toxins such as enzymatically active toxins; growth inhibitors; and drug molecules. The term refers to a substance that has a harmful effect on cell growth or proliferation.

[0119] In this disclosure, the term "small molecule" refers to a small molecule drug that has biological activity.

[0120] In this disclosure, the term "linker" refers to a linker that connects a bioactive molecule to a targeting moiety. It refers to a fragment.

[0121] In this disclosure, the term “targeting portion” refers to a target on the cell surface (or one of the targets). This refers to the portion of a conjugate that can specifically bind to the target. The interaction between the target portion allows for delivery to specific cell populations.

[0122] In this disclosure, if the targeting portion of the conjugate is an antibody, the conjugate This may be referred to as a "drug-antibody conjugate." In this disclosure, "drug-antibody conjugate" "Immune conjugate" and "immune conjugate" are interchangeable.

[0123] In this disclosure, the term "antibody" in its broadest sense refers to antibodies that perform the necessary biological activity. If available, complete monoclonal antibodies, polyclonal antibodies, and at least two complete This is interpreted as including multispecific antibodies (e.g., bispecific antibodies) formed from all antibodies. In this disclosure, the terms “antibody” and “immunoglobulin” are interchangeable.

[0124] In this disclosure, the term “monoclonal antibody” refers to an antibody from a substantially homogeneous group of antibodies. It refers to the body. In other words, with the exception of a few possible natural variations, the antibodies that make up the group are identical. Monoclonal antibodies have high specificity for a single determinant (epitope) of an antigen. However, polyclonal antibodies, which are compared to this, have various determinants (epitopes) It contains antibodies. In addition to specificity, monoclonal antibodies are synthesized by other antibodies. It has the advantage of being uncontaminated. Here, the modifier "monoclonal" means This shows that the antibody is characterized by originating from a substantially homogeneous group of antibodies. It should not be interpreted as being prepared by a special method.

[0125] In some embodiments of this disclosure, monoclonal antibodies also include chimeric antibodies in particular. Well, if the antibody has the required biological activity, then the heavy chain and / or part of the light chain Some antibodies are the same as or identical to a certain type, class, or subclass of antibodies, while the rest are of a different type. , the same as or identical to an antibody of another class or another subclass (e.g., U.S. 4, Issues 816, 567; and Morrison et al., 1984, PNAS, 81:6851-6 See 855). The chimeric antibodies available in this disclosure are from non-human primates (e.g., ancient primates). Contains variable region antigen-binding sequences and human constant region sequences derived from native monkeys or orangutans. It contains primate-like antibodies.

[0126] The term "antibody fragment" refers to a portion of an antibody, preferably the antigen-binding region or the variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, Fd, Fv, dAb, and complementary It includes sex-determining fragments, diabodies, linear antibodies, and single-chain antibody molecules.

[0127] The term "bispecific antibody," also known as "bifunctional antibody conjugate," is used in the context of Formed by the first antibody (fragment) and the second antibody (fragment) via the plucking arm This refers to a conjugate that maintains the activity of each antibody, and therefore bifunctional. It possesses potential and dual specificity properties.

[0128] The term "multispecific antibody" refers to, for example, an antibody that has three different antigen-binding specificities. Triple-specific antibodies, and quadruple-specific antibodies, which have four different antigen-binding specificities. It contains sex antibodies.

[0129] The term "complete antibody" refers to the antigen-binding variable region, the light chain constant region (CL), and the heavy chain constant region. This refers to antibodies containing the regions (CH1, CH2, and CH3). The constant region is the natural sequence (e.g., It may be a human natural constant region sequence or an amino acid sequence variant thereof. Complete antibodies are preferred. Alternatively, it is a complete antibody that has one or more effector functions.

[0130] The term "probody" refers to a group that can specifically bind to its target, with masked bases and cups. A modified antibody containing an antibody or antibody fragment that can be ringed, and the masked group is, here , the cleavage constant (cleavage const) related to the binding ability of an antibody or antibody fragment to a target An antibody (ant) that is not coupled with a masked group to its target, or an antibody that is not bonded to the target. At least 100 times, 1000 times, or 10000 times the cleavage constant for the bonding ability of the piece. It refers to something being high.

[0131] In this disclosure, the "humanization" form of non-human (e.g., mouse) antibodies is described as minimal non-human immunotherapy. This refers to chimeric antibodies containing a robulin sequence. Most humanized antibodies are human recipient antibodies. Residues in the hypervariable region of epidemic globulin possess the necessary specificity, affinity, and function of non-human ( For example, residues in the hypervariable region (donor antibody) of mice, rats, rabbits, or non-human primates. It is replaced by the frame region of human immunoglobulin in some embodiments. The residues in (FR) are also substituted with non-human residues. Furthermore, the humanized antibody is a recipe. It may also contain residues that are not present in the donor antibody or antibody. Such modifications may affect the antibody. Humanized antibodies are created to further optimize physical performance. Humanized antibodies generally have at least one capable A variable region, typically comprising two variable regions, where all or almost all superimposed. The variant loop corresponds to non-human immunoglobulins, but all or almost all FRs are human. It is an immunoglobulin sequence. Humanized antibodies are immunoglobulin constant regions (Fc, normally) It may contain at least a portion of human immunoglobulin (Fc). For details, see For example, Jones et al., 1986, Nature, 321:522~525; Rich mann et al., 1988, Nature, 332:323~329; and Presta, 1 See 992, Curr Op Struct Bwl 2:593-596.

[0132] Complete antibodies can be classified into different "groups" according to the amino acid sequence of the heavy chain constant region. The five groups are IgA, IgD, IgE, IgG, and IgM, and some of them are Also, different types such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2 They can be further classified into subclasses (isotypes). Different groups of the constant region of the heavy chain of antibodies are These are called α, β, ε, γ, and μ, respectively. The subunit structure of immunoglobulins and 3 Various classes of D configurations are well known in this field.

[0133] In this disclosure, amino acid substitutions in antibodies are often substituted with L-amino acids. However, the embodiments are not limited thereto. In some embodiments, the antibody peptide chain may be one or multiple It may contain several D-amino acids. Peptides containing D-amino acids can be found in the oral cavity, intestinal tract, or blood. In plasma, it is more stable and less degradable than peptides containing only L-amino acids. There is.

[0134] The monoclonal antibodies used in this disclosure can be produced by a number of methods. For example, the monoclonal antibodies used in this disclosure are effective against a wide range of species (mice, hamsters, etc.). Hybridoma methods using (including rat and human cells) (e.g., Kohler) See also Nature, 1975, 256:495) or by recombination. This can be obtained by DNA techniques (see, for example, U.S. Patent No. 4,816,567). It can be done, or it can be isolated from a phage antibody library (for example, Cla ckson et al., 1991, Nature, 352:624~628; and Marks et al., 1991, Journal of Molecular Biology, 222:58 See 1-597). Monoclonal antibodies that can be used in this disclosure include: This list includes, but is not limited to, anti-Her2 monoclonal compounds such as trastuzumab and pertuzumab. A nal antibody, or sacituzumab (i.e., isactuzumab or This includes anti-Trop-2 monoclonal antibodies such as hRS7 antibody, M1, M2, or M3. It can be done.

[0135] In some preferred embodiments, the target of A is epidermal growth factor, Trop-2, CD37, HER2, CD70, EGFRvIII, Mesothelin, Folic Acid Folate receptor 1, Mucin 1, CD138, C D20, CD19, CD30, SLTRK6, Nectin4, tissue factor ( Tissue factor), mucin 16, endothelin receptor (En dothelin receptor), STEAP1, SLC39A6, guanylate receptor), STEAP1, SLC39A6, Clase (Guanylylcyclase) C, PSMA, CCD79b, CD22, Sodium phosphate cotransporter ter)2B, GPNMB, Trophoblast glycoprotein (Trophoblast g lycoprotein), AGS-16, EGFR, CD33, CD66e, CD74 , CD56, PD-L1, TACSTD2, DR5, E16, 0772P, MPF, Na pi3b, Sema 5b, PSCA hlg, ETBR, MSG783, STEAP2 TrpM4, CRIPTO, CD21, CD79b, FcRH2, NCA, MDP, I L20Rα レカン(Brevican), EphB2R, ASLG659, PSC A, GEDA, BAFF-R, CD79a, CXCR5, HLA-DOB, P2X5 D72、LY64、FcRH1、IRTA2、TENB2、イテグリン(integr in) α5β6, α4β7, FGF2, FGFR2, Her3, CA6, DLL3, DL L4, P-cadherin, EpCAM, pCAD, CD223, LY PD3, LY6E, EFNA4, ROR1, SLITRK6, 5T4, ENPP3, クロ Claudin (Claudin)18.2, BMPR1B, Tyro7, c-Met, Apo E, CD1 lc, CD40, CD45(PTPRC), CD49D(ITGA4), C D80, CSF1R, CTSD, GZMB, Ly86, MS4A7, PIK3AP1, P.S IK3CD, CCR5, IFNG, IL10RA1, IL-6, ACTA2, COL7A 1. LOX, LRRC15, MCPT8, MMP10, NOG, SERPINEl, ST AT1, TGFBR1, CTSS, PGF, VEGFA, C1QA, C1QB, ANGP TL4, EGLN, EGLN3, BNIP3, AIF1, CCL5, CXCL10, CX CL11, IFI6, PLOD2, KISS1R, STC2, DDIT4, PFKFB3 PGK1, PDK1, AKR1C1, AKR1C2, CADM1, CDH11, COL 6A3, CTGF, HMOX1, KRT33A, LUM, WNT5A, IGFBP3, M MP14, CDCP1, PDGFRA, TCF4, TGF, TGFB1, TGFB2, C.S D1 lb, ADGRE1, EMR2, TNFRSF21, UPK1B, TNFSF9, Choose from MMP16, MFI2, IGF-1R, RNF43, NaPi2b, or BCMA. It will be done.

[0136] In some embodiments of this disclosure, the target of targeting portion A is cell surface integrins. RGD peptides that recognize receptors; cell surface growth factors such as EGF, PDGF, or VEGF. Growth factors that recognize progenitor receptors; and functional cell surface plasminogen activators, Bombe Peptides that can recognize synthin, bradykinin, somatostatin, or prostate-specific membrane antigen receptors Selected from D.

[0137] In some embodiments of this disclosure, the target of targeting portion A is CD40 ligand, C D30 ligand, OX40 ligand, PD-1 ligand, ErbB ligand, Her2 ligand Selected from Gand, TACSTD2 ligand, and DR5 ligand.

[0138] In some embodiments of this disclosure, the targeting portion A is trastuzumab or per Anti-Her2 monoclonal antibodies such as tuzumab; or sacituzumab, M1, M2 or These are anti-Trop-2 monoclonal antibodies such as M3.

[0139] In some embodiments of this disclosure, the targeting portion is trastuzumab or pertuzumab. Trastuzumab is an anti-Her2 monoclonal antibody, and its amino acid sequence is as follows: This information is publicly known to the vendor; for an overview of the sequence, please refer to, for example, CN103319599. .

[0140] In some embodiments of this disclosure, the terminal Lys of the heavy chain of the targeting portion is easily deleted. However, such deletions do not affect biological activity. Dick, LW et al., Bio See technol.Bioeng., 100:1132~1143. For example, The targeting portion is an anti-Trop- such as sacituzumab, which has a deletion of the terminal Lys of the heavy chain. 2 monoclonal antibodies, M1, M2, or M3, for example, the targeting portion is heavy Anti-Her2 monoclonal drugs such as trastuzumab or pertuzumab with deletion of terminal lysine chains. It is a nal antibody.

[0141] Exemplary heavy and light chain sequences of trastuzumab are shown in SEQ ID NOs: 17 and 18. See [reference]. This disclosure refers to the heavy and light chain combinations of trastuzumab referenced or relating to this disclosure. The columns are described using the sequences shown in sequence numbers 17 and 18, respectively. For exemplary heavy and light chain sequences of tuzumab, see U.S. Patent No. 7560111. See 16 and Sequence ID 15.

[0142] Sequence ID 17 (heavy chain sequence) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQ APGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTA YLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPG(K)

[0143] Sequence ID 18 (light chain sequence) DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQK PGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFLTISSLQ PEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

[0144] In some embodiments of this disclosure, the targeting portion of the anti-Trop-2 antibody is a U.S. patent RS7 as described in Patent No. 7,517,964 (i.e., sacituzumab as disclosed herein); and US Patent hRS7 described in Patent Application Publication No. 2012 / 0237518 (i.e., the terms of this disclosure) (Mab) The anti-Trop-2 antibodies available in this disclosure are, by screening, The carrier design, construction, and antibody disclosed in Japanese Patent No. 103476941A are displayed It can also be obtained by constructing a library of antibodies, or by Sorrento T Screening the G-MAB (registered trademark) library from herapeutics, Inc. You can also obtain it by doing so.

[0145] The heavy chain sequence and light chain amino acid sequence of the monoclonal antibody sacituzumab are as follows: For example, see Sequence ID 19 and Sequence ID 20.

[0146] Sequence ID 19 (heavy chain sequence) QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQ APGQGLKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTA YLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG(K)

[0147] The terminal K (or lys) of the heavy chain is readily deleted, but such deletions affect biological activity. It does not affect. Dick, LW et al., Biotechnol. Bioeng., 10 Please refer to 0:1132~1143.

[0148] Sequence ID 20 (light chain sequence) DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQK PGKAPKLLIYSASYRYTGVPDRFSGSGSGTFTLTISSLQ PEDFAVYYCQQHYITPLTFGAGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

[0149] In this disclosure, ErbB2 and Her2 / neu are interchangeable, and both are natural sequences. Human Her2 protein (Genebank CAS number X03363, e.g., Se mba et al., 1985, PNAS, 82:6497~6501; and Yamamoto et al. See Nature, 1986, 319:230-234) and amino acid sequence This refers to functional derivatives such as rianto. ErbB2 is the gene that codes for human Her2. Representing the offspring, neu represents the gene encoding rat p185neu. Some embodiments Therefore, the compounds or conjugates of this disclosure are used in breast cancer cells, ovarian cancer cells, gastric cancer cells, Endometrial cancer cells, salivary gland cancer cells, lung cancer cells, kidney cancer cells, colon cancer cells, thyroid Cancer cells, such as pancreatic cancer cells, bladder cancer cells, or liver cancer cells, express the ErbB2 receptor. It can inhibit or kill cells.

[0150] In this disclosure, Trop-2 or TROP2 refers to TACSTD2, M1S1, GA733 -1, also known as EGP-1, refers to human trophoblast cell surface antigen 2, which is found in many humans. Tumors (e.g., breast cancer, colorectal cancer, lung cancer, pancreatic cancer, ovarian cancer, prostate cancer and children) It is a cell surface receptor expressed in cervical cancer. In some embodiments, the compound of the present disclosure or The conjugate is found in breast cancer cells, colorectal cancer cells, lung cancer cells, pancreatic cancer cells, and ovarian cancer cells. Cells that express the TROP2 receptor, such as cancer cells, prostate cancer cells, or cervical cancer cells. It can be inhibited or killed.

[0151] When used herein, the conjugate of the present invention contains [ka] When the targeting portion is an antibody, the sulfhydryl group and linker in the antibody are involved. This shows a specific linking pattern.

[0152] When used in this specification, "C 1~6 The term "alkyl" is, for example, "C 1~4 a "Lukiru" and "C 1~3 A linear or branched chain containing 1 to 6 carbon atoms, including an alkyl group. This refers to alkyl groups. Specific examples include, but are not limited to, methyl, ethyl, and n-pro groups. Pill, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n -Pentyl, iso-pentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl Nthyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethyl It contains rubyl and 1,2-dimethylpropyl.

[0153] When used in this specification, "C 2~6 The term "Alkenil" is, for example, "C 2~4 A linear chain containing at least one double bond and 2 to 6 carbon atoms, including an "alkenyl". , refers to branched or cyclic alkenyls. 2~6 Examples of "Alkenil" are limited to these. However, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1, 3-Butadienyl, 1-Pentenyl, 2-Pentenyl, 3-Pentenyl, 1,3-Penta Dienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl , 1,4-hexadienyl, cyclopentenyl, 1,3-cyclopentadienyl, cyclo This includes hexenyl and 1,4-cyclohexadienyl.

[0154] When used in this specification, "C 2~6 The term "alkynyl" is, for example, "C 2~4 A linear chain containing at least one triple bond and 2 to 6 carbon atoms, including an "alkynyl" Or it refers to branched alkynyl. 2~6 Examples of "alkinyl" include, but are not limited to, these. Ethinyl, propynyl, 2-butynyl, 2-pentynyl, 3-pentynyl, 4-methyl -2-pentinyl, 2-hexinyl, 3-hexinyl and 5-methyl-2-hexinyl It is included.

[0155] As used herein, the term "halogen" includes fluorine, chlorine, bromine, and iodine. It contains elements.

[0156] When used herein, "3-8 membered cycloalkyl" or "C 3~8 Cycloalkyl The term "cycloalkyl" is used, for example, to refer to "3-6 member cycloalkyl" and "4-6 member cycloalkyl." 3 to 8 carbon atoms containing "5-7 membered cycloalkyl" or "5-6 membered cycloalkyl" This refers to saturated cyclic alkyl groups containing a molecule. Specific examples include, but are not limited to, cyclic alkyl groups. Lopropanyl, cyclobutylalkyl, pentanyl, cyclohexyl, cycloheptyl and It contains bicyclooctadecyl.

[0157] When used in this specification, "C 1~6 The term "alkoxy" is C 1~6 Alkyl- This refers to a group having an O- structure, where C 1~6 Alkyl is defined as previously. Specific examples include, but are not limited to, methoxy, ethoxy, propoxy, and isopropyl compounds. Poxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy It contains toxic, pentoxy, and hexyloxy.

[0158] As used herein, the term "3- to 8-membered aliphatic heterocyclyl" means 3 to 8 The ring-forming atoms (at least one of the ring-forming atoms is a nitrogen atom, an oxygen atom, or a sulfur atom) This refers to a cyclic group containing heteroatoms (such as ).Optionally, the ring-forming element in the cyclic structure. The atom (for example, a carbon atom, a nitrogen atom, or a sulfur atom) may be replaced by oxygen. "8-membered aliphatic heterocyclyls" include, for example, oxyranil, o Xocyclobutyl, pyrrolidinyl, tetrahydrofuranil, piperidinyl, piperazinyl , containing tetrahydropyranil and homopiperazinil, "3-8 member nitrogen-containing aliphatic heterozygotes "Krill", "3-8 member oxygen-containing aliphatic heterocyclyl", "3-6 member aliphatic heterocyclyl "Lu", "3-6 member oxygen-containing aliphatic heterocycline", "4-7 member aliphatic heterocycline" "4-6 member aliphatic heterocyclyl", "5-7 member aliphatic heterocyclyl", "5-6 member This includes "aliphatic heterocyclines" and "5-6 member nitrogen-containing aliphatic heterocyclines."

[0159] As used herein, the term "6-12 member spirocyclyl" means 6-12 member spirocyclyl It contains ring-forming carbon atoms and is formed by two or more cyclic structures sharing one carbon atom. This refers to a cyclic structure. Optionally, carbon atoms in the cyclic structure may be substituted with oxygen. "6-12 member spirocyclils" include, for example, "6-11 member spirocyclils" and "6-1 "0-membered spirocyclyl", "7-10 membered spirocyclyl", "7-9 membered spirocyclyl", "7-8 member spirocyclyl", "9-10 member spirocyclyl", and "3-10 member spirocyclyl This includes "krill". Specific examples are not limited to these, but include: [ka] It includes.

[0160] As used herein, the term "6-12 membered cyclyl" means a ring with 6 to 12 members. Formed by two or more cyclic structures containing a forming carbon atom and sharing two adjacent carbon atoms. This refers to a cyclic structure. Optionally, carbon atoms in the cyclic structure may be substituted with oxygen. "6-12 member bridged cyclils" include, for example, "6-11 member bridged cyclils" and "5-10 "7-10 member cross-linked cyclil", "7-9 member cross-linked cyclil", "7-8 This includes "9-10 member cross-linked cyclyl", "9-10 member cross-linked cyclyl", and "3-10 member cross-linked cyclyl". Examples include, but are not limited to, these. [ka] It includes.

[0161] As used herein, the term "6-12 member condensed cyclyl" means "6-11 member condensed cyclyl". Synthetic cyclyl, 6-10 member condensed cyclyl, 6-8 member condensed cyclyl, 10-12 A ring-forming carbon atom containing 6 to 12 carbon atoms, including a "membered condensed cyclyl" and a "7-10 membered condensed cyclyl". A cyclic structure formed by two or more cyclic structures that contain and share two adjacent atoms. This refers to "6-12 member condensed cyclyls," but examples are not limited to these. [ka] It includes.

[0162] As used herein, the term "6-12 member spiroheterocyril" means 6-1 Two ring-forming carbon atoms (at least one of which is a nitrogen atom, an oxygen atom, or a sulfur atom) It contains heteroatoms such as and has a cyclic structure in which two or more ring-forming atoms share one ring-forming atom. This refers to the cyclic structure formed by this process. Optionally, the ring-forming atoms in the cyclic structure (e.g., carbon) are also included. Atoms (nitrogen atoms or sulfur atoms) may be replaced with oxygen. 6-12 member spirohete For example, "rocyclyl" includes "6-11 member spiroheterocyclyl" and "5-10 member spiro "Heterocyclyl", "7-11 member spiroheterocyclyl", "7-10 member spiroheterocyclyl "Krill", "7-9 member spiroheterocycline", "7-8 member spiroheterocycline", This includes "9-10 member spiroheterocyclines" and "3-10 member spiroheterocyclines". Specific examples include, but are not limited to, [ka] It includes.

[0163] As used herein, the term "6-12 membered cross-linked heterocyclyl" means 6-12 a number of ring-forming atoms (at least one of which is a nitrogen atom, oxygen atom or sulfur atom, etc.) It contains a heteroatom and has a cyclic structure in which two or more non-adjacent ring-forming atoms are shared. This refers to the cyclic structure formed by this process. Optionally, the ring-forming atoms in the cyclic structure (e.g., carbon) are also included. The atoms (nitrogen atoms or sulfur atoms) may be substituted with oxygen. 6-12 membered cross-linked hetero For example, "cyclyl" includes "6-11 member cross-linked heterocyclyl" and "6-9 member cross-linked heterocyclyl." Krill, 6-10 membered cross-linked heterocyclyl, 7-10 membered cross-linked heterocyclyl, "7-9 member cross-linked heterocyclyl", "7-8 member cross-linked heterocyclyl", "8 member cross-linked heterocyclyl "Krill", "9-10 member cross-linked heterocyclyl", and "3-10 member cross-linked heterocyclyl" It is included. Specific examples include, but are not limited to, these. [ka] It includes.

[0164] As used herein, the term "6-12 membered condensed heterocyclyl" means 6-12 a number of ring-forming atoms (at least one of which is a nitrogen atom, oxygen atom or sulfur atom, etc.) It contains heteroatoms and has a cyclic structure in which two or more adjacent atoms are shared. This refers to the cyclic structure formed by the following. Optionally, the ring-forming atoms in the cyclic structure (for example, carbon atoms) are also included. The nitrogen atom or sulfur atom may be replaced with oxygen. 6-12 member condensed heterosulfide "Lil" includes, for example, "6-11 member condensed heterocyclyl" and "5-10 member condensed heterocyclyl" "Lil", "7-10 member condensed heterocyclyl", "3-10 member condensed heterocyclyl", "3 ~10-membered nitrogen-containing condensed heterocyclyl", "9-10 membered condensed heterocyclyl", "9-1 Contains "0-membered nitrogen-containing condensed heterocyclines" and "6-12 membered oxygen-containing condensed heterocyclines". It is found. Specific examples include, but are not limited to, tetrahydroimidazo[4,5-c Pyridyl, 3,4-dihydroquinazolinyl, 1,2-dihydroquinoxalinyl, benzo [d][1,3]dioxolyl, 1,3-dihydroisobenzofuranyl, 4H-1,3- Benzoxazinyl, 4,6-dihydro-1H-fl[3,4-d]imidazolyl, 3a ,4,6,6a-tetrahydro-1H-fl[3,4-d]imidazolyl,4,6-dihy Dro-1H-thieno[3,4-d]imidazolyl, 4,6-dihydro-1H-pyrrolo[3 ,4-d]imidazolyl, benzimidazolidinyl, octahydro-benzo[d]imida Zolyl, Decahydroquinolyl, Hexahydrothienomidazolyl, Hexahydrofloimi Dazolyl, 4,5,6,7-tetrahydro-1H-benzo[d]imidazolyl, octahi Drocyclopentano[c]pyrrolyl, dihydroindolyl, dihydroisoindolyl, Nzooxazolidinyl, benzothiazolidinyl, 1,2,3,4-tetrahydroisoquino Linyl, 1,2,3,4-tetrahydroquinolinyl and 4H-1,3-benzoxazinyl It contains the letter.

[0165] As used herein, the term "aryl" refers to a 6-20 member aryl, a 6-10 member aryl, and a 6-10 member aryl. Aromatic monocyclic or polycyclic hydrocarbols such as 5-8 membered aryls and 5-8 membered aryls It refers to nyl. Specific examples include, but are not limited to, phenyl, naphthyl, and anthracite. This includes senyl and phenanthryl groups. "6-20 membered aryl" groups form 6-20 rings. This refers to an aryl group containing an atom.

[0166] As used herein, the term "heteroaryl" means at least one ring formation. Aromatic cyclic groups whose atoms are heteroatoms such as nitrogen, oxygen, or sulfur atoms. This refers to the ring-forming atoms in the cyclic structure (e.g., carbon atoms, nitrogen atoms, or sulfur atoms). The child may be replaced with oxygen. Specific examples include, but are not limited to, frills. , thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, Isooxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2,3-tri Zolyl, 1,2,4-Triazolyl, 1,2,3-Oxadiazolyl, 1,2,4-Ox Sadiazolyl, 1,2,5-Oxadiazolyl, 1,3,4-Oxadiazolyl, Pyrid Lu, 2-pyridone, 4-pyridone, pyrimidinyl, 1,4-dioxacyclohexadienyl , 2H-1,2-oxazinyl, 4H-1,2-oxazinyl, 6H-1,2-oxazinyl Dinyl, 4H-1,3-oxazinyl, 6H-1,3-oxazinyl, 4H-1,4-oxazinyl Xazazinyl, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl Azinyl, 1,2,4,5-tetradinyl, azacycloheptatrienyl, 1,3-dia 5-10 member heterozygotes such as xacycloheptatrienyl and azacyclooctatetraenyl Reel, 5-10 member nitrogen-containing heteroaryl, 6-10 member oxygen-containing heteroaryl, 6- It contains 8-membered nitrogen-containing heteroaryl compounds and 5- to 8-membered oxygen-containing heteroaryl compounds.

[0167] Beneficial effects of the invention This disclosure relates to a coupling method between the drug and the targeting portion in an ADC or SMDC. By improving the law, we can obtain certain novel bioactive molecule conjugates. In some embodiments of this disclosure, the bioactive molecule conjugate is a free sulfur in the antibody. Obtained by nucleophilic substitution of heteroaryl rings on ADC linkers with hydryls. The conjugate obtained by this coupling has the following technical effects: At least one can be achieved: (1) High stability; (2) High DAR; in some embodiments, the DAR value of the conjugate can reach 5-8. ; (3) Extremely high coupling efficiency; in some embodiments, the coupling efficiency is 90% It can reach; (4) The conjugate obtained by coupling effectively stabilizes the drug in circulation. This can improve the situation and reduce unexpected drug dissociation in non-target cells; (5) Conjugates also increase the effective release of bioactive molecules in cells, thus reducing toxicity. The objectives of reducing [amount] and increasing effectiveness can be achieved; (6) The conjugate has good tumor tissue targeting properties; and (7) Conjugates have high efficacy in animal models of tumors.

[0168] In addition, the coupling method described herein has a broad range of applications, and can be used to convert biologically active molecules into antibodies. Alternatively, it can be widely used when coupling with targeted small molecule ligands. [Brief explanation of the drawing]

[0169] [Figure 1] This shows the TIC (Total Ion Chromatogram) of BT001002. [Figure 2] This shows the deconvolution diagram of the coupled light chain of BT001002. [Figure 3] This shows the deconvolution diagram of the coupled heavy chain of BT001002. [Figure 4] This shows the TIC (Total Ion Chromatogram) of BT001004. [Figure 5] This shows the deconvolution diagram of the coupled light chain of BT001004. [Figure 6] This shows the deconvolution diagram of the coupled heavy chain of BT001004. [Figure 7] This shows the SEC chromatogram for BT001002. [Figure 8] This shows the SEC chromatogram of the molecular weight marker for BT001002. [Figure 9] This shows the SEC chromatogram for BT001004. [Figure 10] This shows the deconvolution diagram of the coupled light chain of BT001012. [Figure 11] This shows the deconvolution diagram of the coupled heavy chain of BT001012. [Figure 12] This shows the deconvolution diagram of the coupled light chain of BT001013. [Figure 13] This shows the deconvolution diagram of the coupled heavy chain of BT001013. [Figure 14] This shows the deconvolution diagram of the coupled light chain of BT001018. [Figure 15] This shows the deconvolution diagram of the coupled heavy chain of BT001018. [Figure 16] This shows the deconvolution diagram of the coupled light chain of BT001021. [Figure 17] This shows the deconvolution diagram of the coupled heavy chain of BT001021. [Figure 18] This shows the deconvolution diagram of the coupled light chain of BT001023. [Figure 19] This shows the deconvolution diagram of the coupled heavy chain of BT001023. [Figure 20] This shows the deconvolution diagram of the coupled light chain of BT001040. [Figure 21] This shows the deconvolution diagram of the coupled heavy chain of BT001040. [Figure 22] This shows the deconvolution diagram of the coupled light chain of BT001041. [Figure 23] This shows the deconvolution diagram of the coupled heavy chain of BT001041. [Figure 24] This shows the deconvolution diagram of the coupled light chain of BT001042. [Figure 25] This shows the deconvolution diagram of the coupled heavy chain of BT001042. [Figure 26] This shows the deconvolution diagram of the coupled light chain of BT001043. [Figure 27] This shows the deconvolution diagram of the coupled heavy chain of BT001043. [Figure 28] This shows the deconvolution diagram of the coupled light chain of BT001044. [Figure 29] This shows the deconvolution diagram of the coupled heavy chain of BT001044. [Figure 30] This shows the deconvolution diagram of the coupled light chain of BT001046. [Figure 31] This shows the deconvolution diagram of the coupled heavy chain of BT001046. [Figure 32] This shows the deconvolution diagram of the coupled light chain of BT001047. [Figure 33] This shows the deconvolution diagram of the coupled heavy chain of BT001047. [Figure 34] This shows the SEC chromatogram for BT001012. [Figure 35] This shows the SEC chromatogram of BT001013. [Figure 36] This shows the SEC chromatogram of BT001018. [Figure 37] This shows the SEC chromatogram for BT001021. [Figure 38] This shows the SEC chromatogram for BT001023. [Figure 39] This shows the SEC chromatogram of BT001042. [Figure 40] This shows the SEC chromatogram of BT001043. [Figure 41] This shows the SEC chromatogram of BT001044. [Figure 42] This shows the SEC chromatogram for BT001046. [Figure 43] This shows the SEC chromatogram for BT001047. [Figure 44] The changes in tumor growth volume in each mouse group in the NCI-N87 human gastric cancer model are shown. [Figure 45] The weight changes of each mouse group in the NCI-N87 human gastric cancer model are shown. [Figure 46] The changes in tumor growth volume in each mouse group in the HCC1806 human breast cancer model are shown. [Figure 47A] The changes in tumor growth volume in each mouse group in a xenograft model of HCC827 human non-small cell lung cancer are shown. [Figure 47B] The weight changes in each mouse group in a xenograft model of HCC827 human non-small cell lung cancer are shown. [Figure 48A] The changes in tumor growth volume in each mouse group in a xenograft model of NCI-N87 human gastric cancer are shown. [Figure 48B] The weight changes of each mouse group in the NCI-N87 human gastric cancer xenograft model are shown. [Figure 49A] The changes in tumor growth volume in each mouse group in the MDA-MB-231 human breast cancer tumor-bearing mouse model are shown. [Figure 49B] The weight changes of each mouse group in the MDA-MB-231 human breast cancer tumor-bearing mouse model are shown. Specific methods for carrying out the invention

[0170] This disclosure will be further illustrated in combination with specific embodiments, but this disclosure is not an embodiment of those embodiments. Not limited to the present state. Those skilled in the art will not deviate from the basic idea and scope of this disclosure. It should be understood that various changes or improvements may be made in accordance with the teachings in this disclosure.

[0171] The abbreviations used in this invention have the following meanings: [Table 2]

[0172] Preparation solution The structure of the compound described in the following example is subjected to nuclear magnetic resonance ( 1 H NMR) or mass spectrometry (MS ) was decided by.

[0173] Nuclear magnetic resonance ( 1 ¹H NMR is performed using a Bruker 400MHz NMR spectrometer. This was determined by: Deuterated methanol (CD3OD), Deuterated chloroform ( The solvent for determination is either CDCl3 or deuterated dimethyl sulfoxide (DMSO-D6). Therefore, tetramethylsilane (TMS) was used as the internal standard.

[0174] The abbreviations for nuclear magnetic resonance (NMR) spectra used in the examples are shown below. s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quartet double line, ddd: double double double line, ddt: double double triple line, dddd: double double double double line Multiline, m: multiple lines, br: broad, J: coupling constant, Hz: Hertz, DMSO-d6: heavy Dimethyl hydrogenated sulfoxide. δ value is expressed in ppm.

[0175] Mass spectrum (MS) obtained from Agilent (ESI) mass spectrometer (model: Agil The determination was made using ent 6120B).

[0176] Preparative liquid chromatography: Method A: Chromatography column: Daisogel C18 10μm 100×25 0mm Mobile phase A: Water; Mobile phase B: Acetonitrile [Table 3]

[0177] Method B: Chromatography column: Daisogel C18 10μm 50×250 mm Mobile phase A: Water; Mobile phase B: Acetonitrile [Table 4]

[0178] Method C: Chromatography column: Daisogel C18 10μm 50×250 mm Mobile phase A: Water containing 0.1% trifluoroacetic acid; Mobile phase B: Acetonitrile [Table 5]

[0179] Method D: Chromatography column: Waters SunFire C18 5μm 1 9 x 250 mm Mobile phase A: Acetonitrile; Mobile phase B: Water containing 0.05% formic acid Time: 0min~16min; Mobile phase A: 10%~90%; Flow rate: 28mL / min

[0180] I. Synthesis of Bioactive Molecules Example 1: (2S)-N-((3R,4S,5S)-1-((2S)-2-((1R, 2R)-3-((1-((4-aminobenzyl)amino)-1-oxo-3-phenyl Ropan-2-yl)amino)-1-methoxy-2-methyl-3-propionyl)pyrrolidine (1-yl)-3-methoxy-5-methyl-1-heptanoyl-4-yl)-2-(( S)-2-(dimethylamino)-3-methylbutyrylamino)-N,3-dimethylbutyl Synthesis of amide (T001) [ka]

[0181] Step 1: tert-butyl(4-((2-((2R,3R)-3-((S)-1 -((3R,4S,5S)-4-((S)-2-((S)-2-(dimethylamino)-3 -methylbutyrylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5methyl (Heptanoyl)pyrroridine-2-yl)-3-methoxy-2-methylpropionamide Synthesis of )-3-phenylpropionamide)methyl)phenyl)carbamate 1-hydroxybenzotriazole (2.0 mg, 14.74 μmol) was incubated at room temperature in N, Dissolve in N-dimethylformamide (4 mL), cool to 0°C, then tert-butyl 4-Methylaminobenzylcarbamate (4.0 mg, 16.1 μmol), N,N-di Isopropylethylamine (8.5 mg, 66.8 μmol), ((2R, 3R)-3- ((S)-1-((3R,4S,5S)-4-((S)-2-((S)-2-(dimethyl Amino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino)-3-meth Xy-5-methylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methyl Lopionyl-L-phenylalanine (10.0 mg, 13.5 μmol, commercially available) administered sequentially It was added to the target. After stirring for 5 minutes, 1H-benzotriazole-1-yl-oxytripy Loridinophosphonium hexafluorophosphate (10.0 mg, 20.1 μmol) The mixture was added to it and stirred at 0°C for 1 hour. The reaction of the starting materials was observed using high-performance liquid chromatography. The reaction was monitored by quantitative analysis. After the raw materials were consumed, the reaction solution was separated by preparative liquid chromatography. The title compound (9.0 mg of a white solid) was obtained by purification using Fee (Method D). ESI -MS(m / z):950.5[M+H] + .

[0182] Step 2: (2S)-N-((3R,4S,5S)-1-((2S)-2-((1 R,2R)-3-((1-((4-aminobenzyl)amino)-1-oxo-3-pheny Lupropan-2-yl)amino)-1-methoxy-2-methyl-3-propionyl)pyro Lysine-1-yl)-3-methoxy-5-methyl-1-heptanoyl-4-yl)-2- ((S)-2-(dimethylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino Synthesis of tanamides tert-butyl(4-((2-((2R,3R)-3-((S)-1-((3 R,4S,5S)-4-((S)-2-((S)-2-(dimethylamino)-3-methyl Butyrylamino)-N,3-dimethylbutyryl)-3-methoxy-5-methylheptanoyl (Pyrrolidine-2-yl)-3-methoxy-2-methylpropionyl)-3-phenyl Propanamide (methyl)phenyl)carbamate (9.0 mg, 0.02 mmol) Dissolve in 1,4-dioxane (0.5 mL), cool to 0°C, and then the salt in the dioxane... Hydrogenated hydrogen solution (1 mL, 4.0 M) was added, and the mixture was reacted at room temperature for 3 hours with stirring. The response was monitored by high-performance liquid chromatography-mass spectrometry. The raw materials were consumed. Next, the solvent is evaporated under reduced pressure, and the crude product is obtained by preparative liquid chromatography (Method C). The compound was purified to obtain the trifluoroacetate salt of the title compound (5.0 mg of a white solid). ESI -MS(m / z):850.5[M+H] + .

[0183] Example 2: (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R )-3-((S)-1-((4-aminobenzyl)amino)1-oxo-3-phenyl Ropa-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine (-1-yl)-3-methoxy-5-methyl-1-oxoheptyl-4-yl)-2-( (S)-2-(dimethylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino Synthesis of Luamide (T011) [ka]

[0184] Step 1: tert-butchi(S)-(4-((2-(((((9H-fluoren- 9-yl)methoxy)carbonyl)amino)-3-phenylpropionamide)methyl) Synthesis of phenyl(L)carbamate At 0°C, 4-aminobenzylamine (222 mg, 1.0 mmol) and N-methylmethylamine Ruforin (306 mg, 1.5 mmol) in N,N-dimethylformamide (5 mL) (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino) Add to a solution of -3-phenylpropionic acid (387 mg, 1.0 mmol), then 1 -Hydroxybenzotriazole (203 mg, 1.5 mmol) and 1-(3-dimethicone) (288 mg, 1.5 mg) 3-ethylcarbodiimide hydrochloride (288 mg, 1.5 mg) (mol) was added continuously. The resulting mixture was reacted overnight at 0°C. The reaction solution was then diluted with water. When poured into 50 mL of water, a white solid precipitated. The solid was filtered, and the filter cake was placed in water (2 mL). Washed with 0 mL x 3). The solid was purified by silica gel column chromatography. The title compound (380 mg of a white solid) was obtained. ESI-MS (m / z): 592.3[M +H] + .

[0185] Step 2: tert-butyl(S)-(4-((2-amino-3-phenylpropyl Synthesis of ionamide methyl phenyl carbamate Dissolve lithium hydroxide monohydrate (21 mg, 0.51 mmol) in water (1 mL) and t ert-butyl(S)-(4-((2-((((9H-fluorene-9-yl)methoxy )carbonyl)amino)-3-phenylpropionamide)methyl)phenyl)carbamyl) A solution of tetrahydrofuran (2 mL) containing 102 mg of tetrahydrofuran (0.17 mmol) was added to the solution. The resulting mixture was reacted at room temperature for 2 hours. Water (20 mL) was added to the reaction solution, and acetic acid was added. Extraction was performed with ethyl acetate (30 mL x 4). The organic phase was combined with saturated physiological saline (30 mL x 2). The material was washed and dried on anhydrous sodium sulfate. The drying agent was then removed by filtration, and the solvent was removed. The solution is evaporated under reduced pressure, and the residue is purified by preparative liquid chromatography (Method D). The title compound (65 mg of a white solid) was obtained. ESI-MS (m / z): 370.2[M+ H] + .

[0186] Step 3: (4-((S)-2-((2R,3R)-3-((S)-1-((3R ,4S,5S)-4-((S)-3-((methylamino)-3-methylbutyrylamino) -N,3-dimethylbutyrylamino)-3-methoxy-5-methylheptanoyl) pylori (Zin-2-yl)-3-methoxy-2-methylpropionamide)-3-phenylpropionamide Synthesis of ionamide methyl phenyl carbamate At 0°C, tert-butyl(S)-(4-((2-amino-3-phenylpropionate Mid(methyl)phenyl)carbamate (15 mg, 0.04 mmol) and N-methyl Morpholine (12 mg, 0.12 mmol) in N,N-dimethylformamide (2 mL) (2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)-2- ((S)-2-(dimethylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino (Tyrylamino)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl-3- Add to a solution of methoxy-2-methylpropionic acid (24 mg, 0.04 mmol), then 1-hydroxybenzotriazole (8 mg, 0.06 mmol) and 1-(3-di Methylaminopropyl)-3-ethylcarbodiimide hydrochloride (12 mg, 0.0 6 mmol) was added continuously. The resulting mixture was reacted overnight at 0°C. The reaction solution The title compound (white solid 24m) was purified by preparative liquid chromatography (Method D). g) was obtained. ESI-MS (m / z): 950.6 [M+H] + .

[0187] Step 4: (S)-N-((3R,4S,5S)-1-((S)-2-((1R, 2R)-3-((S)-1-((4-aminobenzyl)amino)-1-oxo-3-fe Nylpropa-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl) (Loridine-1-yl)-3-methoxy-5-methyl-1-oxoheptyl-4-yl)- 2-((S)-2-(dimethylamino)-3-methylbutyrylamino)-N,3-dimethyl Synthesis of butylamide Add trifluoroacetic acid (0.5 mL) to dichloromethane (1.5 mL) (4-((S) -2-((2R,3R)-3-((S)-1-(3R,4S,5S)-4-((S)-3 -(methylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino) -3-Methoxy-5-methylheptanoyl)pyrroridine-2-yl)-3-Methoxy-2 -Methylpropanamide)-3-phenylpropionamide)methyl)phenyl)carb It was added to a solution of Maat (14.0 mg, 0.015 mmol). The resulting mixture was stored at room temperature. The reaction was allowed to proceed for 1 hour. Then, the solvent was evaporated under reduced pressure, and the residue was collected by preparative liquid chromatography. The trifluoroacetate of the title compound was purified by method C (a white solid). mg was obtained. ESI-MS (m / z): 850.6 [M+H] + .

[0188] The following molecules can be synthesized by a similar synthetic method. [Table 6]

[0189] Example 3: (S)-N-(2-(4-ethyl-4-hydroxyl-3,14-dione- 3,4,12,14-tetrahydro-1H-pyrano[3',4',6,7]indo[1, 2-b] Synthesis of quinoline-11-yl)ethyl)-N-isopropylacetamide [ka]

[0190] Berotecan hydrochloride (1.0g, 2.13 mmol) and triethylamine (0.65g, Dissolve 0.9 mL of dichloromethane (50 mL) at room temperature, and add acetic anhydride (0.22 g, 2 (0.13 mmol) was slowly added dropwise. The resulting mixture was reacted at room temperature for 1 hour. The organic phase was washed with water (10 mL x 2) and dried on anhydrous sodium sulfate. Insoluble substances Remove by filtration, evaporate the solvent, and the residue is subjected to silica gel column chromatography (di The title compound (1 g) was obtained by purification using chloromethane / methanol = 50 / 1). ESI-MS (m / z): 476.2 [M+H] + .

[0191] Example 4: (S)-N-(2-(4-ethyl-4-hydroxyl-3,14-dione -3,4,12,14-tetrahydro-1H-pyrano[3',4',6,7]indridi No[1,2-b]quinoline-11-yl(ethyl)-N-isopropylmethanesulfone Mido synthesis [ka]

[0192] Methylsulfonyl chloride (462 mg, 12.77 mmol, purity: approximately 70%) is dichloromethylsulfonyl chloride. berotecan hydrochloride (3g, 6.38 mmol) and triethyl alcohol in lomethane (40 mL) It was added dropwise to a solution of mine (2.58 g, 25.54 mmol). The resulting mixture was left at room temperature. The mixture was allowed to react for 2 hours. The mixture was then filtered by suction, and the filter cake was washed three times with dichloromethane (3 mL). After purification, the title compound (2.2g) was obtained.

[0193] The structural characteristics data is as follows: 1 H NMR(400MHz,DMSO-d6)δ8.32(d,J=8.4Hz,1 H), 8.20(dd,J=8.4,1.2Hz,1H), 7.93~7.84(m,1 H), 7.79(t,J=7.6Hz,1H), 7.35(s,1H), 6.56(s, 1H), 5.44(d,J=9.2Hz,4H), 3.98(p,J=6.7Hz,1H ), 3.50(t,J=8.0Hz,2H), 3.42~3.35(m,2H), 3.0 0(s,3H), 1.93~1.82(m,2H), 1.15(d,J=6.7Hz,6 H), 0.88(t,J=7.3Hz,3H). ESI-MS(m / z):512.2[ M+H] + . [α]D 20 is +28.19°(c=0.101g / 100mL,CH3C N)

[0194] The remaining bioactive molecules whose synthesis methods are not described are either commercially available or are based on prior art. It can be prepared by the method of disclosure.

[0195] II. Synthesis of Bioactive Molecules and Linker-Containing Compounds Example 5: (S)-2-((S)-2-(4-(4-chloro-7H-pyrrolo[2,3- d] Pyrimidine-7-yl)butyrylamide)-3-methylbutyrylamide)-N-(4 -(((S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4 -((S)-2-((S)-2-(dimethylamino)-3-methylbutyrylamino)-N ,3-dimethylbutyrylamino)-3-methoxy-5-methylheptanoyl)pyrrolidine (-2-yl)-3-methoxy-2-methylpropionamide)-3-phenylpropion Synthesis of amide)methyl)phenyl)-5-ureidobarrelamide [ka]

[0196] Step 1: tert-butyl4-(4-chloro-7H-pyrrolo[2,3-d] Synthesis of Midine-7-yl)butyrate (compounds 1-2) At room temperature, compound 1-1 (500 mg, 3.27 mmol) was dissolved in N,N-dimethylforma Dissolve in 10 mL of iodine, and add sodium hydride (130 mg, 3.27 mmol) to the solution. It was slowly added in batches. The resulting mixture was stirred at room temperature for 10 minutes, followed by 4-B 725 mg of t-butyl romobylate (3.27 mmol) was added dropwise, and then at room temperature for 2 hours. The reaction was allowed to proceed. The reaction was quenched with saturated ammonium chloride aqueous solution, and ethyl acetate (50 mL) was added. Extraction was performed using (3 x 3). Next, the organic phase was combined and washed with saturated physiological saline (50 mL x 3). The material was dried on anhydrous sodium sulfate. The drying agent was removed by filtration, and the solvent was evaporated under reduced pressure. Then, the title compound (500 mg) was obtained. ESI-MS (m / z): 296.1[M+H ] + .

[0197] Step 2: 4-(4-chloro-7H-pyrrolo[2,3-d]pyrimidine-7-yl) Synthesis of butyric acid (compounds 1-3) At room temperature, compound 1-2 (500 mg, 1.69 mmol) was dissolved in dichloromethane (6 mL). It was dissolved in [a solution], trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 4 hours. Then, it was dissolved in [another solution]. The medium was evaporated under reduced pressure to obtain the title compound (400 mg). ESI-MS (m / z): 240.1[M+H] + .

[0198] Step 3: (9H-fluoren-9-yl)-methyl-((S)-1-(((S)- 1-((4-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino )-1-oxo-5-ureidopentyl-2-yl)amino)-3-methyl-1-oxo Synthesis of butyl-2-yl)-carbamates (compounds 1-5) 4-(N-Boc-aminomethyl)-aniline (6.0 g, 27 mmol) at room temperature, Compound 1-4 (3.35g, 6.75mmol) and 2-ethioxyl-1-ethoxycal Boxyl-1,2-dihydroquinoline (3.34g, 13.5mmol) is sterilized with dichlorometh Dissolve in a mixed solvent of 140 mL of ethanol and 70 mL of methanol, then heat at 45°C. It was heated and reacted at that temperature for 8.0 hours. After cooling to room temperature, a large amount of solid precipitated, and The mixture was subjected to suction filtration to obtain the title compound (3.65 g). ESI-MS (m / z): 70 1.4[M+H] + .

[0199] Step 4: (9H-fluoren-9-yl)-methyl-((S)-1-(((S) -1-((4-(aminomethyl)phenyl))amino)-1-oxo-5-5-ureido Pentyl-2-yl)amino)-3-methyl-1-oxobutyl-2-yl)carbama Synthesis of compounds (1-6) At room temperature, add trifluoroacetic acid (15 mL) to compound 1-5 (3.0 g, 4.29 mmol). It was added and stirred at room temperature for 1.0 hour. Then the solvent was evaporated under reduced pressure to obtain a yellow oily substance. The result was obtained. Upon adding anhydrous diethyl ether (20 mL), a large amount of solid precipitated. 0. After vigorous stirring for 5 hours, suction filtration was performed to obtain the trifluoroacetate (3) of the title compound. 0.6g was obtained. ESI-MS (m / z): 601.3 [M+H] + .

[0200] Step 5: (9H-fluoren-9-yl)-methyl-((S)-1-(((S) -1-((4-(((R)-2-((t-butyloxycarboryl)amino)-3-fe Nylpropionamide)methyl)phenyl)amino-1-oxo-5-ureidopentyl -2-yl)amino)-3-methyl-1-oxobutyl-2-yl)-carbamate(chemical Synthesis of compound 1-7) Boc-D-phenylalanine (1.1 g, 4.2 mmol) and compound 1-6 at room temperature The trifluoroacetate (3.0g, 4.2 mmol) is mixed with N,N-dimethylformamide ( Dissolve in 40 mL, cool to 0°C, then 1-(3-dimethylaminopropyl)-3- Ethylcarbodiimide hydrochloride (1.2g, 6.3mmol), 1-hydroxybe Nzotriazole (0.9g, 6.3mmol) and N-methylmorpholine (1.7g, 16.8 mmol was added continuously. The reaction system was stirred at that temperature for 1.0 hour. Next Then, the reaction solution is added dropwise to ice water (400 mL) and stirred vigorously for 0.5 hours, and a large amount A solid precipitate formed. The title compound (3.3g) was obtained by suction filtration. ESI-MS (m / z):848.4[M+H] + .

[0201] Step 6: (9H-fluoren-9-yl)-methyl-((S)-1-(((S) -1-((4-(((R)-2-amino-3-phenylpropionamide)methyl)fe Nyl)amino)-1-oxo-5-ureidopentyl-2-yl)amino)-3-methyl Synthesis of -1-oxobutyl-2-yl)-carbamates (compounds 1-8) At room temperature, dissolve compounds 1-7 (3.0 g, 3.3 mmol) in trifluoroacetic acid (30 mL). The solution was dissolved and stirred at room temperature for 1.0 hour. The solvent was evaporated under reduced pressure to obtain a yellow oily substance. Adding 100 mL of water-diethyl ether and stirring vigorously for 0.5 hours results in a large amount of solid. A precipitate formed. The trifluoroacetate salt (2.1 g) of the title compound was obtained by suction filtration. ESI-MS (m / z): 748.4 [M+H] + .

[0202] Step 7: (9H-fluoren-9-yl)-methyl-((S)-1-(((S) -1-((4-(((S)-2-((2R,3R)-3-((S)-1-((3R,4S ,5S)-4-((S)-2-((S)-2-(dimethylamino)-3-butyrylamide )-N,3-dimethylamino)-3-methoxy-5-methylheptanoyl)pyro-2-y (Lu)-3-methoxy-2-methylpropionamide)-3-phenylpropionamide) Methyl(phenyl)amino)-1-oxo-5-ureidopenta-2-yl)amino)- Synthesis of 3-methyl-1-oxobutan-2-yl)carbamate (compounds 1-9) (2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)- 2-((S)-(dimethylamino)-3-butyrylamino)-N,3-dimethylbutyryl Amino)-3-methoxy-5-methylheptanoyl)pyro-2-yl)-3-methoxy- 2-Methylpropionic acid (1.3g, 2.17mmol) and trifluor of compounds 1-8 1.8g, 2.17 mmol of bisacetate in N,N-dimethylformamide (20 mL) Dissolve in [a solution], cool to 0°C, then 1-hydroxybenzotriazole (440 mg, 3. (26 mmol) and N-methylmorpholine (658 mg, 6.51 mmol) administered sequentially. Add and finally 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydro Chloride (624 mg, 1.38 mmol) was added. The reaction solution was stirred at 0°C for 5 hours. The title compound (1.8g) was purified by preparative liquid chromatography (Method D). Got it. ESI-MS(m / z):1329.2[M+H] + .

[0203] Step 8: (S)-2-((S)-2-amino-3-butyrylamino)-N-(4 -(((S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4 -((S)-2-((S)-2-(dimethylamino)-3-butyrylamino)-N,3- Dimethylbutyrylamino)-3-methoxy-5-methylheptanoyl)pyro-2-yl) (-3-Methoxy-2-methylpropionamide)-3-phenylpropionamide)Meth Synthesis of phenyl-5-ureidobarrelamide (compounds 1-10) At room temperature, compounds 1-9 (500 mg, 0.38 mmol) were dissolved in N,N-dimethylforma. Dissolve in mid (5 mL), add piperidine (324 mg, 3.8 mmol), and at room temperature. The mixture was stirred for 3 hours. Then, purification was performed by preparative liquid chromatography (Method D) to obtain the title compound. A substance (350 mg) was obtained. ESI-MS (m / z): 1107.2 [M+H] + .

[0204] Step 9: (S)-2-((S)-2-(4-(4-chloro-7H-pyrrolo[2, 3-d]Pyrimidine-7-yl)butyrylamide)-3-methylbutyrylamide)-N- (4-(((S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S) -4-((S)-2-((S)-2-(dimethylamino)-3-methylbutyrylamino) -N,3-dimethylbutyrylamino)-3-methoxy-5-methylheptanoyl) pylori (Zin-2-yl)-3-methoxy-2-methylpropionamide)-3-phenylpropionamide Compound 5-Ureidobarrelamide (compound TL001) Growth Compounds 1-10 (60 mg, 0.054 mmol) and 4-(4-chloro-7H) are collected at room temperature. -Pyrrolo[2,3-d]pyrimidine-7-yl)butyrate (26 mg, 0.066 mmol) Dissolve in N,N-dimethylformamide (3 mL), cool to 0°C, and N,N-diisopropyl alcohol. Ropyrethylamine (105 mg, 0.81 mmol) and 1H-benzotriazole- 1-Oxytripyrrolidinophosphonium hexafluorophosphate (281 mg, 0. 54 mmol was added continuously. The reaction system was stirred at room temperature for 3 hours. Then the purified solution was separated. The title compound (30 mg) was obtained by liquid chromatography (Method D). ESI-M S(m / z): 664.5[M / 2+H] + .

[0205] Example 6: (S)-N-(4-(((S)-2-((2R,3R)-3-((S)-1 -((3R,4S,5S)-4-((S)-2-((S)-2-(dimethylamino)-3 -methylbutyrylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5-methylbutyrylamino) (Tylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropionamide (d)-3-phenylpropionamide)methyl)phenyl)-2-((S)-3-methyl -2-(4-(4-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidine-7 -yl)-butyrylamide)-butyrylamide)-5-ureidobarrelamide [ka]

[0206] Step 1: 4-(4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine- Synthesis of 7-yl)butyric acid (compound 2-2) At room temperature, 4-(4-chloro-7H-pyrrolo[2,3-d]pyrimidine-7-yl)butyrate Dissolve (300 mg, 1.25 mmol) in methanol (8 mL) and sodium methane Thiol (351 mg, 5.02 mmol) is added in one dose, and then the mixture is heated to 50°C. The reaction was carried out overnight. Purification was performed by preparative liquid chromatography (Method D) to obtain the title compound (1 20 mg was obtained. ESI-MS (m / z): 252.1 [M+H] + .

[0207] Step 2: (S)-N-(4-(((S)-2-((2R,3R)-3-((S) -1-((3R,4S,5S)-4-((S)-2-((S)-2-(dimethylamino) -3-methylbutyrylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5 -Methylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropion (amide)-3-phenylpropionamide)methyl)phenyl)-2-((S)-3-methyl Tyl-2-(4-(4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine- 7-Iyl)-Butyrylamide)-Butyrylamide)-5-Ureidobarrelamide (compound) 2-3) Synthesis 4-(4-chloro-7H-pyrrolo[2,3-d]pyrimidine-7-yl)butyrate 4-( Replaced with 4-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-7-yl)butyric acid Except for the fact that the procedure was carried out in the same manner as described in step 9 of Example 5, the purification was performed. The title compound (20 mg) was obtained using preparative liquid chromatography (Method D). ESI-MS (m / z): 670.5 [M / 2 + H] + .

[0208] Step 3: (S)-N-(4-(((S)-2-((2R,3R)-3-((S) -1-((3R,4S,5S)-4-((S)-2-((S)-2-(dimethylamino) -3-methylbutyrylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5 -Methylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropion (amide)-3-phenylpropionamide)methyl)phenyl)-2-((S)-3-methyl Tyl-2-(4-(4-(methylsulfonyl)-7H-pyrrolo[2,3-d]pyrimidine -7-yl)-butyrylamide)-butyrylamide)-5-ureidobarrelamide(compound) Synthesis of material TL002) At room temperature, compound 2-3 (20 mg, 0.015 mmol) was dissolved in dichloromethane (2 mL). Dissolve in [a solution] and add m-chloroperoxybenzoic acid (4.0 mg, 0.022 mmol). The resulting mixture was reacted at room temperature for 2 hours. Purification was performed by preparative liquid chromatography (method The title compound (5.0 mg) was obtained by method D). ESI-MS (m / z): 686.5 [M / 2+H] + .

[0209] Example 7: N-((S)-1-(((S)-1-((4-(((S)-2-((2R, 3R)-3-((S)-1-((3R,4S,5S)-4-((S)-2-((S)-2 -(dimethylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino )-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3-methoxy- 2-Methylpropionamide-3-phenylpropionamide)methyl)phenyl)ami (no)-1-oxo-5-ureido-2-yl)amino)-3-methyl-1-oxybutane -2-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)-5-hexine Amido [ka]

[0210] Step 1: Methyl 6-(2-(methylthio)pyrimidine-5-yl)-5-hexyl Synthesis of Noart (compound 3-2) At room temperature, methyl 5-hexinoate (500 mg, 3.97 mmol) and 5-bromo -2-methylthiopyrimidine is dissolved in N,N-dimethylformamide (3 ml), and then The solution contains triethylamine (3 ml), copper iodide (75 mg, 0.4 mmol), and bis(tri) Phenylphosphine palladium(II) dichloride (279 mg, 0.4 mmol) The mixture was added continuously. The resulting mixture was heated to 95°C under nitrogen protection and reacted with stirring for 6 hours. The solution was quenched with water and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and saturated physiological solution was extracted. Washed with saline solution (20 mL x 2) and dried on anhydrous sodium sulfate. The desiccant was filtered. The residue was removed, and the solvent was evaporated under reduced pressure. Purification was performed by preparative liquid chromatography (Method D). The title compound (300 mg) was obtained. ESI-MS (m / z): 251.3 [M+H ] + .

[0211] Step 2: 6-(2-(methylthio)pyrimidine-5-yl)-5-hexic acid ( Synthesis of compound 3-3) At room temperature, compound 3-2 (200 mg, 0.8 mmol) is dissolved in tetrahydrofuran and water ( Dissolve in a 4 mL / 4 mL mixed solution of lithium hydroxide monohydrate (235 mg, 5.6 mL). Add (mol), stir, and react at room temperature for 4 hours, then dilute with water and ethyl acetate ( Extraction was performed using 20 ml x 2. The aqueous phase was adjusted to pH=3 with 1N hydrochloric acid, and ethyl acetate (20 ml) was used. Extract with ×3), then combine the organic phase and wash with saturated physiological saline (20 mL × 2), It was dried on anhydrous sodium sulfate. The desiccant was removed by filtration, and the solvent was evaporated under reduced pressure. Then, the title compound (120 mg) was obtained.

[0212] Step 3: 6-(2-(methylsulfonyl)pyrimidine-5-yl)-5-hexy Synthesis of nic acid (compounds 3-4) At room temperature, compound 3-3 (20 mg, 0.085 mmol) is dissolved in dichloromethane (4 mL). Dissolve in [the solution], add m-chloroperoxybenzoic acid (22 mg, 0.127 mmol), The reaction was carried out overnight at room temperature under stirring. Purification was performed by preparative liquid chromatography (Method D). The title compound (20 mg) was obtained. ESI-MS(m / z):269.1[M+H] + .

[0213] Step 4: N-((S)-1-(((S)-1-((4-(((S)-2-((2 R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)-2-((S) -2-(dimethylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamino Mino)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3-methoxy C-2-methylpropionamide)-3-phenylpropionamide)methyl)phenyl )amino)-1-oxo-5-ureido-2-yl)amino)-3-methyl-1-oxy Butan-2-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)-5-yl Kixinamide (compound TL003) 4-(4-chloro-7H-pyrrolo[2,3-d]pyrimidine-7-yl)butyrate 6-( The substitution was made with 2-(methylsulfonyl)pyrimidine-5-yl)-5-hexic acid. Except for the above, the same procedure as described in step 9 of Example 5 was carried out. Purification was performed using a preparative liquid. The title compound (14m) was obtained using chromatography (Method D). ESI-MS (m / z): 679.0 [M / 2 + H] + .

[0214] Example 8: (S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamide (d)-ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyra No[3',4',6,7]-Indrizino[1,2-b]-Quinoline-4-yl(4-( (S)-42-(2-(methylsulfonyl)pyrimidine-5-yl)-4,8,37-to Rioxo-2-(3-ureidopropyl)-6,12,15,18,21,24,27, 30,33-Nonoxy-3,9,36-Azatetracosan-41-amide)benzyl) Carbonato [ka]

[0215] Step 1: Methyl(S)-(1-((4-(hydroxymethyl)phenyl)amino )-1-oxo-5-ureidopentane-2-yl)-(9H-fluorenyl)carbama Synthesis of compound 19-2 Fmoc-L-citrulline (5.0g, 12.58 mmol), p-aminobene at room temperature Diyl alcohol (6.20g, 50.32mmol) and 2-ethoxy-1-ethoxycarbon Ruboxyl-1,2-dihydroquinoline (6.22g, 25.16mmol) is chloro The solution was dissolved in methane (100 mL), heated to 45°C, and reacted for 6 hours. The reaction solution was then cooled under reduced pressure. Concentrate with , stir with anhydrous diethyl ether (100 mL), and the title compound (6.0 g) Got it. ESI-MS(m / z):503.3[M+H] + .

[0216] Step 2: (S)-2-amino-N-(4-(hydroxymethyl)phenyl)-5 - Synthesis of ureidobarrelamide (compound 19-3) Compound 19-2 (1.0 g, 1.99 mmol) was dissolved in N,N-dimethylformamine at room temperature. Dissolve in (8 mL) and steep at room temperature for 30 minutes in a solution containing piperidine (339 mg, 3.98 mmol). The mixture was added dropwise and allowed to react. Then dichloromethane (10 mL) was added, followed by 10 minutes. The mixture was stirred. The reaction solution was concentrated under reduced pressure and analyzed by flash column chromatography. The compound was purified to obtain the title compound (400 mg). ESI-MS (m / z): 281.2[M +H] + .

[0217] Step 3: (S)-2-(32-azid-5-oxo-3,9,12,15,18 ,21,24,27,30-nonyloxa-6-azatriacetamide)-N-(4-( Synthesis of hydroxymethyl)phenyl)-5-ureidobarrelamide (compound 19-4) Compound 19-3 (150 mg, 0.54 mmol) and 32-azido-5-oxo-3 ,9,12,15,18,21,24,27,30-nonoxy-6-azatricylo Dissolve ndecanoic acid (296 mg, 0.54 mmol) in dichloromethane (10 mL), Cool to 0°C, then 2-ethoxy-1-ethoxycarboxyl-1,2-dihydroquinone Phosphorus (145 mg, 0.58 mmol) was added. The resulting mixture was allowed to rise to room temperature and incubated overnight. The reaction solution was then concentrated under reduced pressure and purified by flash column chromatography. The compound was prepared to obtain the title compound (200 mg). ESI-MS (m / z): 817.5[M+ H] + .

[0218] Step 4: 4-((S)-35-Azid-4,8-Dioxo-2-(3-Ureid Propyl)-6,12,15,18,21,24,27,30,33-nonoxy-3, 9-Azatetracosane)benzyl((S)-4-ethyl-11-(2-(N-isopropyl Methylsulfonylamino(ethyl)-3,14-dioxo-3,4,12,14-teto Lahydro-1H-pyrano[3',4',6,7]indridino[1,2-b]quinoline- Synthesis of 4-yl)carbonate (compound 19-5) At room temperature, (S)-N-(2-(4-ethyl-4-hydroxy-3,14-dione-3,4 ,12,14-tetrahydro-1H-pyrano[3',4',6,7]indolidino[1, 2-b]Quinoline-11-yl)ethyl)-N-isopropylmethanesulfonamide(2 Dissolve 00 mg (0.39 mmol) in dichloromethane (10 mL), cool to 0°C, 4-dimethylaminopyridine (573 mg, 4.69 mg) in dichloromethane (1.0 ml) Add a solution of mmol) and then add triphosgene (1) in dichloromethane (1.0 ml). A 16 mg (0.39 mmol) solution was slowly added dropwise. The resulting mixture was then stirred. Below, the reaction was carried out at 0°C for 1 hour. Compound 19-4 (15) in dichloromethane (2.0 mL) A 9 mg (0.18 mmol) solution was added to the reaction solution and reacted at room temperature for 1 hour. Purification The title compound (160 mg) was obtained by preparative high-performance liquid chromatography (Method D). ESI-MS (m / z): 678.0 [M / 2 + H] + .

[0219] Step 5: 4-((S)-35-amino-4,8-dioxo-2-(3-ureido Propyl)-6,12,15,18,21,24,27,30,33-nonoxy-3, 9-Azatetracosane)benzyl((S)-4-ethyl-11-(2-(N-isopropyl Methylsulfonylamino(ethyl)-3,14-dioxo-3,4,12,14-teto Lahydro-1H-pyrano[3',4',6,7]indridino[1,2-b]quinoline- Synthesis of 4-yl)carbonate (compound 19-6) Compound 19-5 (80 mg, 0.059 mmol) was dissolved at room temperature in tetrahydrofuran (1. Dissolve in (0 ml), cool to 0°C, then add 4-dichloromethane (1.0 ml) Add a solution of thylaminopyridine (573 mg, 4.69 mmol) and platinum dioxide (1 Add 5 mg (0.059 mmol) in one batch under nitrogen protection, then add air and hydrogen. The reaction was carried out three times by substitution and allowed to proceed at room temperature for 6 hours. The reaction solution was filtered, and the filtrate was concentrated to obtain the crude product. This was obtained. This was purified by preparative high-performance liquid chromatography (Method D) to obtain the title compound. (40 mg) was obtained. ESI-MS (m / z): 665.0 [M / 2 + H] + .

[0220] Step 6: (S)-4-ethyl-11-(2-(N-isopropylmethylsulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4',6,7]Indrizino[1,2-b]Quinoline-4-yl(4-(( S)-42-(2-(methylsulfonyl)pyrimidine-5-yl)-4,8,37-tri Oxo-2-(3-ureidopropyl)-6,12,15,18,21,24,27,3 0,33-nonoxy-3,9,36-azatetracosan-41-amide)benzyl) Synthesis of lubonate (compound TL019) Compound 19-6 (30 mg, 0.016 mmol) and 6-(2-methylsulfonyl Limidine-5-yl)-5-hexic acid (6.4 mg, 0.024 mmol) N,N- Dissolve in dimethylformamide (1 mL), cool to 0°C, then benzotriazole- 1-Iloxytripyrrolidinyl hexafluorophosphate (16.5 mg, 0.0 32 mmol), N,N-diisopropylethylamine (6.2 mg, 0.047 mmol) l) was added continuously. The resulting mixture was reacted at room temperature for 2 hours. The purified solution was then separated and extracted at high performance. The title compound (10 mg) was obtained by chromatography (Method D). ESI-MS (m / z): 790.0 [M / 2 + H] + .

[0221] Example 9: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfonamide (I)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3',4',6,7]Indrizino[1,2-b]Quinoline-4-yl-(4-((S )-2-((S)-3-methyl-2-(6-(2-(methylsulfonyl)pyrimidine-5 -Il))-5-hexinamide)butylamide-5-ureidovalerylamide)benzyl Carbonato [ka]

[0222] Step 1: Methyl((S)-1-(((S)-1-((4-(hydroxymethyl) Phenyl)amino)-1-oxo-5-ureidovalerylamido-2-yl)amino)) (-3-methyl-butylamido-2-yl)-(9H-fluorenyl)carbamate Step 1 of Example 8 is the same except that compound 19-1 is replaced with compound 28-1. The title compound (310 mg) was obtained by performing the same procedure as described. ESI-MS ( m / z): 602.3[M+H] + .

[0223] Step 2: (S)-2-((S)-2-amino-3-methylbutylamide)-N- (4-(hydroxymethyl)phenyl)-5-ureidovaleramide (compound 28-2) synthesis Step 2 of Example 8 was followed, except that compound 19-2 was replaced with compound 28-2. The title compound (150 mg) was obtained by performing the same procedure as described. ESI-MS ( m / z): 380.3 [M+H] + .

[0224] Step 3: N-((S)-1-(((S)-1-((4-hydroxymethyl)fe Nyl)amino)-1-oxo-5-ureidopenta-2-yl)amino)-3-methyl- 1-Oxobutan-2-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl) Synthesis of )-5-hexinamide (compound 28-4) Benzotriazole-1-yloxytripyrrolidinyl hexafluorophosph at room temperature Art (313 mg, 0.6 mmol) and N,N-diisopropylethylamine (19 4 mg (1.50 mmol) in N,N-dimethylformamide (10 mL) 6-(2 -Methylsulfonylpyrimidine-5-yl)-5-hexic acid (135 mg, 0.5 mm ol) and (2S)-2-(((2S)-2-amino-3-methyl-butyryl)amino) -N-(4-(hydroxymethyl)phenyl)-5-ureido-valleamide(190m The reaction solution was added to a 0.5 mmol solution and stirred at room temperature for 3 hours. The title compound (78 mg) was obtained by purification using high-performance liquid chromatography (Method D). ESI-MS (m / z): 630.3 [M+H] + .

[0225] Step 4: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4',6,7]Indrizino[1,2-b]Quinoline-4-yl-(4-( (S)-2-((S)-3-methyl-2-(6-(2-(methylsulfonyl)pyrimidine -5-yl))-5-hexinamide)butylamide-5-ureidovalerylamide) Synthesis of dyl carbonate (compound TL028) Step 4 of Example 8 was carried out with the exception that compound 19-4 was replaced with compound 28-4. The title compound (1.76 mg) was obtained by performing the same procedure as described. ESI-MS (m / z): 1167.4[M+H] + .

[0226] Example 10: (S)-4-ethyl-11-(2-(N-isopropylmethylsulfonate Mido(ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyra No[3',4',6,7]Indrizino[1,2-b]Quinoline-4-yl-(4-(( 2S,5S)-5-isopropyl-38-(4-((6-(2-(methylsulfonyl) Limidine-5-yl)-5-hexinamide)methyl)-1H-1,2,3-triazole Ru-1-yl)-4,7,11-trioxo-2-(3-ureidopropyl)-9,15 ,18,21,24,27,30,33,36-nonoxy-3,6,12-triazoto Riacontylamide (benzyl)carbonate [ka]

[0227] Step 1: (S)-2-((S)-35-azido-2-isopropyl-4,8-di Oxo-6,12,15,18,21,24,27,30,33-Nonoxy-3,9- Azatetracosyl)-N-(4-(hydroxymethyl)phenyl)-5-ureidovaler Synthesis of amides (compound 29-1) Step 3 of Example 8 was carried out with the exception that compound 19-3 was replaced with compound 28-3. The title compound (180 mg) was obtained by performing the same procedure as described. ESI-MS ( m / z): 916.5 [M+H] + .

[0228] Step 2: 4-((2S,5S)-38-azido-5-isopropyl-4,7,1 1-Trioxo-2-(3-ureidopropyl)-9,15,18,21,24,27, 30,33,36-nonoxy-3,6,12-triazatriacontylamide)benzyl Lu((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamide)eth (Lu)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3', [4',6,7]Indolidino[1,2-b]Quinoline-4-yl)carbonate (compound) Synthesis of 29-2) Step 4 of Example 8 was carried out with the exception that compound 19-4 was replaced with compound 29-1. The title compound (30 mg) was obtained by performing the same procedure as described. ESI-MS (m / z):727.5[M / 2+H]+ .

[0229] Step 3: (S)-4-ethyl-11-(2-(N-isopropylmethylsulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4',6,7]Indrizino[1,2-b]Quinoline-4-yl-(4-( (2S,5S)-5-isopropyl-38-(4-((6-(2-(methylsulfonyl) Pyrimidine-5-yl)-5-hexinamide)methyl)-1H-1,2,3-triazo (L-1-yl)-4,7,11-trioxo-2-(3-ureidopropyl)-9,1 5,18,21,24,27,30,33,36-nonoxy-3,6,12-triazo Synthesis of triacontylamide benzyl carbonate (compound TL029) At room temperature, compound 29-2 (20 mg, 0.014 mmol) and 6-(2-(methyls Rufonyl)pyrimidine-5-yl)-N-(2-propyne n-1-yl)-5-hexine Amid (4.3 mg, 0.014 mmol) in a mixed solvent of dimethyl sulfoxide and water ( Dissolve in 1 mL / 0.25 mL, then add copper bromide (3.95 mg, 0.027 mmol). ) was added and the mixture was reacted under stirring for 1 hour. Purification was performed by preparative high-performance liquid chromatography (Method D). The title compound (15 mg) was obtained by ESI-MS (m / z): 880.0 [M / 2+H] + .

[0230] Example 11: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfonate Mido(ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyra No[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-((2 S,5S)-5-isopropyl-45-(2-(methylsulfonyl)pyrimidine-5-I Lu)-4,7,11,40-tetraoxo-2-(3-ureidopropyl)-9,15, 18,21,24,27,30,33,36-nonoxy-3,6,12,39-tetra Azapentatetracontane-44-carbamoyl(benzyl)carbonate [ka]

[0231] Step 1: (S)-2-((S)-35-amino-2-isopropyl-4,8-di Oxo-6,12,15,18,21,24,27,30,33-Nonoxy-3,9- Diazapentatriacontoamide)-N-(4-(hydroxymethyl)phenyl)-5- Synthesis of ureidobarrelamide Compound 29-1 (400 mg, 0.44 mmol) is dissolved in methanol and tetrahydrochloride at 20°C. It was dissolved in Drofuran (2.0 mL:4.0 mL). After dissolution was complete, platinum dioxide (40 mL) was added. g) was added in one batch under nitrogen protection, and the mixed solution was then subjected to hydrogen exchange three times. The reaction was carried out at 20°C for 2 hours. The reaction solution was filtered. The filtered cake was washed with methanol. The filtrate was concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Method D). The title compound (200 mg) was prepared. ESI-MS (m / z): 890.4[M+H ] + .

[0232] Step 2: N-((6S,9S)-1-amino-6-((4-(hydroxymethyl )phenyl)carbamoyl)-9-isopropyl-1,8,11,15-tetraoxo- 13,19,22,25,28,31,34,37,40-nonoxy-2,7,10, 16-Tetraazadotetraconto-42-yl)-6-(2-(methylsulfonyl)pyri Synthesis of Midine-5-yl)hexa-5-inylamide At 20°C, compound 22-1 (250 mg, 0.28 mmol) was dissolved in N,N-dimethylform Dissolve in Muamido (1.0 mL), then add HATU (160 mg, 0.42 mmol) and Then, N,N-diisopropylethylamine (109 mg, 0.84 mmol) is added sequentially. The mixture was then stirred overnight at room temperature. Purification was performed by preparative high-performance liquid chromatography (Method D). The title compound (250 mg) was obtained.

[0233] Step 3: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-(( 2S,5S)-5-isopropyl-45-(2-(methylsulfonyl)pyrimidine-5- Il)-4,7,11,40-tetraoxo-2-(3-ureidopropyl)-9,15 ,18,21,24,27,30,33,36-nonoxy-3,6,12,39-teto Laazapentatetracontane-44-carbamoyl(benzyl)carbonate(compound T Synthesis of L022) (S)-N-(2-(4-ethyl-4-hydroxy-3,14-dione-3, 4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indridino[1 ,2-b]Quinoline-11-yl)ethyl)-N-isopropylmethanesulfonamide( Dissolve 70 mg (0.14 mmol) in dichloromethane (4.0 mL) and cool to 0°C. Next, p-dimethylaminopyridine (200 mg) in dichloromethane (1.0 ml) Add a solution of 1.64 mmol of triphosgene in dichloromethane (1.0 ml). A solution of (40.6 mg, 0.14 mmol) was slowly added dropwise. The resulting mixture The mixture was reacted at 0°C for 1 hour under stirring. Nitrogen was blown into the unreacted triphosgene, and dichlorohydrate was added. React with a solution of compound 22-2 (139 mg, 0.12 mmol) in methane (2.0 mL). The solution was added and reacted with stirring at 0°C for 1 hour. The purified solution was then subjected to preparative high-performance liquid chromatography. The title compound (1.5 mg) was obtained using method D. ESI-MS (m / z): 83 9.5 [M / 2+H] + .

[0234] Example 12: 4-((S)-2-(4-aminobutyl)-42-(2-(methylsulfon Nyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12,15,18,2 1,24,27,30,33-nonanoxa-3,9,36-triazadedotetracontyl -41-Alkynamide)benzyl-((S)-4-ethyl-11-(2-(N-isopropyl Ropyrmethylsulfonyl)ethyl)-3,14-dioxo-3,4,12,14-tetra Hydro-1H-pyrone[3',4':6,7]indolidino[1,2-b]quinoline-4 -Il Carbonato [ka]

[0235] Step 1: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Ron[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-(( S)-2-(4-(((4-methoxyphenyl)benzhydryl)amino)butyl)-4 2-(2-(methylsulfonyl)pyrimidine-5-yl)-4,8,37-trioxo- 6,12,15,18,21,24,27,30,33-nononaoxa-3,9,36-to Synthesis of riazadotetracontyl-41-alkynamide)benzylcarbonate At room temperature, 6-(-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-yne Dissolve the acid (12 mg, 0.045 mmol) in dichloromethane (2 mL), then 2 -(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexaful Olophosphat (21.2 mg, 0.056 mmol) and N,N-diisopropyl alcohol Add thylamine (8.6 mg, 0.067 mmol), stir for 10 minutes, and compound 24 -1 (35 mg, 0.022 mmol) was added and the mixture was reacted with stirring for 1 hour. The purified solution was then separated. The title compound (20 mg) was obtained by high-performance liquid chromatography (Method B). ESI -MS(m / z):1821.8[M+H] + .

[0236] Step 2: 4-((S)-2-(4-aminobutyl)-42-(2-(methylsulfur Honyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12,15,18, 21,24,27,30,33-nonanoxa-3,9,36-triazadedotetraconti L-41-alkynamide)benzyl-((S)-4-ethyl-11-(2-(N-iso Propylmethylsulfonyl(ethyl)-3,14-dioxo-3,4,12,14-teto Lahydro-1H-pyrone[3',4':6,7]indolidino[1,2-b]quinoline- Synthesis of 4-yl)carbonate (compound TL024) At room temperature, compound 24-2 (20 mg, 0.011 mmol) was dissolved in acetonitrile (1 mL). Dissolve in (0.5 ml) trifluoroacetic acid (0.5 ml) in acetonitrile (0.5 ml) and steep for 20 minutes. The solution from (1) was added dropwise and stirred. Purification was performed by preparative high-performance liquid chromatography (Method C). The trifluoroacetate salt (12 mg) of the title compound was obtained using ESI-MS (m / z). :1549.6[M+H] + .

[0237] Example 13: (S)-4-ethyl-11-(2-(N-isopropylmethylsulfonate Mido(ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyra No[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl-4-((2 S,5S)-5-isopropyl-2-methyl-38-(4-((6-(2-(methylsulfur Honyl)pyrimidine-5-yl)yl)hexa-5-inamide)methyl)-1H-1, 2,3-Triazole-1-yl)-4,7,11-Trioxo-9,15,18,21 ,24,27,30,33,36-nonoxy-3,6,12-triazatriacontour Synthesis of Zanamide (benzyl)carbonate [ka]

[0238] Step 1: (S)-(9H-fluoren-9-yl)-methyl(1-((4-(H) Droxymethyl(phenyl(amino)-1-oxopropyl-2-yl)carbamate preparation At room temperature, 2-ethoxy-1-ethoxycarboxyl-1,2-dihydroquinoline (1. 31g, 5.30 mmol) and p-aminobenzyl alcohol (593mg, 4.82 ( mmol) compound 30-1 (1.5 g, 4.82 ml) in dichloromethane (35 mL) The solution was added to (ol) and reacted under stirring for 3 hours. Purification was performed by silica gel column chromatography. The title compound (1.8g) was obtained by ESI-MS (m / z): 417.2 M+H] +

[0239] Step 2: (S)-2-amino-N-(4-(hydroxymethyl)phenyl)pro Preparation of pionamide At room temperature, ethylenediamine (5 mL) is dissolved in dichloromethane (20 mL) with compound 30- It was added to solution 2 (1.8g, 4.32 mmol) and reacted for 2 hours. Purification was performed using silica gel. The title compound (820 mg) was obtained by 2-column chromatography. ESI-MS ( m / z): 195.1[M+H]+

[0240] Step 3: (9H-fluoren-9-yl)-methyl((S)-1-(((S)- 1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl Preparation of amino)-3-methyl-1-oxobutan-2-yl)carbamate At room temperature, (2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)- 3-Methylbutyrate (875 mg, 2.58 mmol), O-benzotriazolyl-tetra Methyluronium hexafluorophosphate (1.45 g, 3.83 mmol), N, N-diisopropylethylamine (1.00 g, 7.74 mmol) and 1-hydroxy Benzotriazole (525 mg, 3.89 mmol) in dichloromethane (2 mL) The compound 30-3 (503 mg, 2.58 mmol) was continuously added to the solution and stirred for 4 hours. The reaction was carried out. Purification was performed by silica gel column chromatography to obtain the title compound (1.1 g) was obtained. ESI-MS (m / z): 516.2[M+H]+

[0241] Step 4: (S)-2-amino-N-((S)-1-((4-(hydroxymethyl Preparation of phenyl)amino)-1-oxopropa-2-yl)-3-methylbutanamide At room temperature, ethylenediamine (2 mL) is dissolved in dichloromethane (8 mL) in compound 30-4 (1.1g, 2.13 mmol) was added to a solution and reacted with stirring for 1 hour. Purification was performed using silicone. The title compound (610 mg) was obtained by Kagel column chromatography. ESI-M S(m / z):294.2[M+H] +

[0242] Step 5: (S)-2-(32-azid-5-oxo-3,9,12,15,18 ,21,24,27,30-nonoxy-6-diazapentatriacontoamide)-N- ((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropane Preparation of -2-yl)-3-methylbutylamide At room temperature, O-benzotriazolyl-tetramethyluronium hexafluorophosph (160 mg, 0.42 mmol), 1-hydroxybenzotriazole (57 mg, 0.42 mmol), N,N-diisopropylethylamine (109 mg, 0.84 mg) ol) and 32-azido-5-oxo-3,9,12,15,18,21,24,27, 30-Nonoxy-6-Azatricyclodecano-1-Acid (156 mg, 0.28 mmol) ) dissolved in dichloromethane (3 mL) compound 30-5 (84 mg, 0.28 mmol) The solution was added and reacted under stirring for 4 hours. Purification was performed by silica gel column chromatography. The title compound (163 mg) was obtained. ESI-MS (m / z): 830.4 [M+H] +

[0243] Step 6: 4-((2S,5S)-38 azido-5-isopropyl-2-methyl- 4,7,11-Trioxo-9,15,18,21,24,27,30,33,36-No ((S)-4-ethyl) -11-(2-(N-isopropylmethylsulfonylamino)ethyl)-3,14-diode Kiso-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]india Preparation of Lisino[1,2-b]quinoline-4-yl)carbonate Under nitrogen protection, at 0°C, triphosgene (16 mg, 0.0) is found in dichloromethane (0.3 mL). A solution of 4-dimethylaminopyridine (0.5 mmol) in dichloromethane (0.7 mL) (65 mg, 0.53 mmol) and (S)-N-(2-(4-ethyl-4-hydroxy -3,14-Dioxo-3,4,12,14-Tetrahydro-1H-Pyrano[3',4' :6,7]Indolidino[1,2-b]quinoline-11-yl)ethyl)-N-isopropyl Add dropwise to a mixed solution of pyrmethanesulfonamide (45 mg, 0.09 mmol), 0 The reaction was carried out at °C for 1 hour. Then, compound 30-6 (73m) was reacted in dichloromethane (1 mL). A solution of (g, 0.09 mmol) was added dropwise to the reaction solution and reacted at 0°C for 1 hour. The title compound (33 mg) was obtained by silica gel column chromatography. ESI -MS(m / z):1367.6[M+H] +

[0244] Step 7: (S)-4-ethyl-11-(2-(N-isopropylmethylsulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4':6,7Indrizino[1,2-b]Quinoline-4-yl-4-((2 S,5S)-5-isopropyl-2-methyl-38-(4-((6-(2-(methylsulfur Honyl)pyrimidine-5-yl)yl)hexa-5-inamide)methyl)-1H-1, 2,3-Triazole-1-yl)-4,7,11-Trioxo-9,15,18,21 ,24,27,30,33,36-nonoxy-3,6,12-triazatriacontour Preparation of Zanamide benzylcarbonate (compound TL030) At room temperature, copper bromide (5 mg, 0.04 mmol) and compound 30-7 (20 mg, 15 µg) (mol) in water and N,N-dimethylformamide (0.2 ml:0.8 ml) 6- (2-(methylsulfonyl)pyrimidine-5-yl)-N-(propa-2-in-1-yl) Add dropwise to a solution of (Lu)-hexa-5-inylamide (9 mg, 0.007 mmol), The mixture was reacted under stirring for 4 hours. Purification was performed by preparative high-performance liquid chromatography (Method D) and labeled. The title compound (4.15 mg) was obtained. ESI-MS(m / z):1672.7[M+H] +

[0245] Example 14: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2- (Methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)methyl)-1H -1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18, 21,24,27,30,33-nonoxa-3,9-diazapentatriacontoamide )benzyl((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamide (I)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl)carbonate [ka]

[0246] Step 1: 6-(2-(methylsulfonyl)pyrimidine-5-yl)-N-(pro Synthesis of P-2-in-1-yl)hexa-5-inamide At 25℃, propa-2-inyl-1-amine (189 mg, 3.4 mmol) and compound Dissolve substance 3-4 (800 mg, 2.83 mmol) in dichloromethane (10 mL), and then N,N-diisopropylethylamine (738 mg, 5.67 mmol) and O-( 7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluroni Muhexafluorophosphate (1.63 g, 4.25 mmol) was added continuously and stirred. The mixture was stirred and reacted for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was collected in a flash silica gel column. Purified by chromatography (ethyl acetate / petroleum ether = 3 / 1), the title compound A substance (700 mg) was obtained. ESI-MS (m / z): 306.1 [M+H] + .

[0247] Step 2: 4-((S)-35-azido-2-(4-(((4-methoxyphenyl )Benzhydryl)amino)butyl)-4,8-dioxo-6,12,15,18,21 ,24,27,30,33-non-azo-3,9-diazapentatricontoamino)ben Zyl((S)-4-ethyl-11-(2-(N-isopropylmethanesulfonamide) (Chill)-3,14-dioxo-3,4,12,14-tetrahydro-2H-pyrano[2, 3-b]-1H-Pyrano[3',4':6,7]Indrizino[1,2-b]Quinoline- Synthesis of 4-yl)carbonate Under nitrogen protection, T-030 (250 mg, 0.49 mmol) is dissolved in dichloromethane at 25°C. Dissolve in (10 mL), cool to 0°C, then 4-dimethicone in dichloromethane (3 mL) Add a solution of diaminopyridine (478 mg, 3.91 mmol), followed by dichloro Slowly dissolve triphosgene (72 mg, 0.24 mmol) in methane (10 mL). The mixture was added dropwise and reacted with stirring at 0°C for 20 minutes. Nitrogen was blown into the reaction solution for 20 minutes. Next, (S)-2-(32-azido-5-oxo-3, in dichloromethane (7 mL) 9,12,15,18,21,24,27,30-nonoxa-6-azatriacetami (d)-N-(4-(hydroxymethyl)phenyl)-6(((4-methoxyphenyl)be Add a solution of acetamide (518 mg, 0.49 mmol) with hydrylaminoacetamide. The reaction was carried out at 0°C for 1 hour under stirring. The reaction solution was concentrated under reduced pressure, and the residue was separated into high-performance liquid crystals. The title compound (500 mg) was obtained by purification using chromatography (Method A). ES I-MS (m / z): 1597.5 [M+H] + .

[0248] Step 3: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-(( S)-2-(4-(((4-methoxyphenyl)diphenylmethyl)amino)butyl)- 35-(4-((6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5- (Inamide)methyl)-1H-1,2,3-triazol-1-yl)-4,8-dioxy So-6,12,15,18,21,24,27,30,33-nonoxy-3,9-dia Synthesis of Zapenta Triacontoamide (benzyl) Carbonate At room temperature, compound 33-1 (14 mg, 0.05 mmol) is dissolved in dimethyl sulfoxide and Dissolve in water (2.0 mL:0.5 mL), then add copper bromide (11 mg, 0.08 mmol) ) was added and the mixture was reacted under stirring for 1 hour. Purification was performed by preparative high-performance liquid chromatography (Method B). The title compound (30 mg) was obtained by ESI-MS (m / z): 815.9[(M (-273) / 2+H] + .

[0249] Step 4: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2 -(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)methyl)-1 H-1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18 ,21,24,27,30,33-nonoxa-3,9-diazapentatriacontoami (D)benzyl((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonate Mido(ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyra [3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl)Carbonate Synthesis of compound TL033 Compound 33-2 (30 mg, 0.02 mmol) was dissolved in dichloromethane (1.0 mL). Dissolve the mixture, add trifluoroacetic acid (0.2 mL) to the reaction solution, and allow to react at room temperature for 30 minutes. The trifluorocarbon of the title compound was purified by preparative high-performance liquid chromatography (Method C). Acetate (20.0 mg) was obtained. The identification of the title compound is as follows: 1 H NMR(400MHz,DMSO-d6)δ10.18(s,1H), 9.10 (s,2H), 8.38(t,J=5.56Hz,1H), 8.32(d,J=8.40 Hz,1H), 8.22~8.20(m,2H), 8.09(t,J=5.68Hz,1 H), 7.91~7.87(m,2H), 7.82~7.78(m,1H),7.69( brs,3H), 7.61(d,J=8.56Hz,2H), 7.32(d,J=8.5 6Hz,2H), 7.06(s,1H), 5.56(d,J=16.96Hz,1H), 5.51(d,J=16.96Hz,1H), 5.47(d,J=19.28Hz,1H ), 5.42(d,J=19.28Hz,1H), 5.14(d,J=12.20Hz, 1H), 5.07(d,J=12.16Hz,1H), 4.48(t,J=5.24Hz ,2H), 4.46~4.43(m,1H), 4.29(d,J=5.60Hz,2H) , 4.08~3.95(m,5H), 3.79(t,J=5.28Hz,2H), 3.5 1~3.43(m,32H), 3.40(s,3H), 3.39~3.35(m,2H) , 3.30~3.26(m,2H), 3.00(s,3H), 2.82~2.74(m, 2H), 2.56(t,J=7.08Hz,2H), 2.29(t,J=7.36Hz, 2H), 2.23~2.13(m,2H), 1.82(p,J=7.24Hz,2H), 1.78~1.63(m,2H), 1.61~1.49(m,2H), 1.42~1.2 7(m,2H), 1.15(d,J=6.80Hz,3H), 1.13(d,J=6.7 6Hz,3H), 0.90(t,J=7.32Hz,3H). ESI-MS(m / z): 816.0[M / 2+H] + . [α] D 20 is -19.55°(c=1.000g / 10 It is 0 mL (CH3CN).

[0250] Example 15: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2- (Methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)methyl)-1H -1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18, 21,24,27,30,33-nonoxy-3,9-diazapentatriacontoamide )benzyl((S)-11-diethyl-9-hydroxy-3,14-dioxo-3,4, 12,14-Tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2 -b]Quinoline-4-carbonate [ka]

[0251] Step 1: 4-((S)-35-azido-2-(4-(((4-methoxyphenyl )diphenylmethyl)amino)butyl)-4,8-dioxo6,12,15,18,21 ,24,27-nonoxy-((S)-9-((tert-butyldimethylsilyl)oxy C)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro- 1,2,3,4-Tetrahydroquinoline-1H-pyrano[3',4':6,7]indri Synthesis of dino[1,2-b]quinoline-4-yl)carbonate Compound 34-1 (100 mg, 0.2 mmol) was dissolved in anhydrous dichloromethane (2 ml) at room temperature. Dissolve in ) under nitrogen protection, then cool to 0°C, followed by anhydrous dichloromethane (0.5 ml) Add the solution of 4-dimethylaminopyridine (144 mg, 1.18 mmol) inside, then Triphosgene (41 mg, 0.14 mmol) in anhydrous dichloromethane (0.5 ml) The solution of ) was slowly added dropwise. The resulting mixture was reacted with stirring at 0°C for 1 hour. Next, (S)-2-(32-azido-5-o) in anhydrous dichloromethane (0.5 mL) Kiso-3,9,12,15,18,21,24,27,30-nonanoxa-6-azatri Acetamide)-N-(4-(hydroxymethyl)phenyl)-6(((4-Methoxymethyl) Dissolved in phenyl(benzhydryl)amino(acetamide) (160 mg, 0.15 μmol) The solution was added to the reaction solution and reacted at room temperature for 1 hour. Purification was performed by preparative high-performance liquid chromatography. The title compound (60 mg) was obtained using method B. ESI-MS (m / z): 159 2.7[M+H] + .

[0252] Step 2: (S)-9-(tert-butyldimethylsilyl)oxy)-4,11 -Diethyl-3,14-Dioxo-3,4,12,14-Tetrahydro-1H-Pyrano[ 3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl-4-((S)- 2-(4-(((6-2-(methylsulfonyl)pyrimidine-5-yl)-35-(4- ((6-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide) (Tyl)-1H-1,2,3-triazole-1-yl)-dioxo6,12,15,18 ,21,24,27,30,33-nonoxy-3,9-diazapentatriacontoami (D) Synthesis of carbonate At room temperature, compound 34-2 (40 mg, 0.03 mmol) and 6-(2-(methylsulfur Honyl)pyrimidine-5-yl)-N-(propa-2-in-1-yl)hexa-5-yl Nylamide (11.50 mg, 0.04 mmol) is mixed with dimethyl sulfoxide and water (0. Dissolve in 5 ml (0.1 ml) and add copper bromide (9.01 mg, 0.06 mmol). The resulting mixture was reacted under stirring for 1 hour. Purification was performed by preparative high-performance liquid chromatography ( The title compound (20 mg) was obtained using method B). ESI-MS (m / z): 1897. 5[M+H].

[0253] Step 3: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2 -(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)methyl)-1 H-1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18 ,21,24,27,30,33-nonoxy-3,9-diazapentatriacontoami (D)benzyl((S)-11-diethyl-9-hydroxy-3,14-dioxo-3,4 ,12,14-tetrahydro-1H-pyrano[3',4':6,7]indridino[1, 2-b] Synthesis of quinoline-4-carbonate (compound TL034) At room temperature, compound 34-3 (30 mg, 0.018 mmol) is dissolved in acetonitrile and water ( Dissolve in 0.4 mL:0.1 mL, then add trifluoroacetic acid and acetonitrile (0 A mixed solution of 0.5 mL and 0.5 mL was added dropwise, and the mixture was stirred at room temperature for 2 hours. Purified solution was then separated into portions. The trifluoroacetate of the title compound (12m) was obtained by fast liquid chromatography (Method C). g) was obtained. ESI-MS (m / z): 1511.5 [M+H] + .

[0254] Example 16: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2- (Methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)methyl)-1H -1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18, 21,24,27,30,33-nonoxa-3,9-diazapentatriacontoamide )benzyl((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamide (I)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl)carbonate synthesis [ka]

[0255] Step 1: ((S)-35-azido-2-(4-(((4-methoxyphenyl)di Phenylmethyl)amino)butyl)-4,8-dioxo-6,12,15,18,21, 24,27,30,33-nonoxy-3,9-diazapentatriacontoamide)ben Zyl((S)-4-ethyl-11-(2-(N-isopropylacetamido)ethyl)- 3,14-Dioxo-3,4,12,14-Tetrahydro-1,2,3,6-Triazaci Chloheptane-1H-Pyrano[3',4':6,7]Indrizino[1,2-b]Quinori Synthesis of 4-yl carbonate Step 1 of Example 15, except that compound 34-1 was replaced with compound 35-1. The title compound (60 mg) was obtained by performing the same procedure as described above. ESI-MS ( m / z): 1561.5 [M+H] + .

[0256] Step 2: (S)-4-ethyl-11-(2-(N-isopropylacetamide) Ethyl)-3,14-dioxo3,4,12,14-tetrahydro-1H-pyrano[3' ,4':6,7]Indrizino[1,2-b]Quinoline-4-yl)-4-((S)-2 -(4-(((4-methoxyphenyl)diphenylmethyl)amino)butyl)-35-( 4-((6-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide )methyl)-1H-1,2,3-triazol-1-yl)-4,8-dioxo-6,1 2,15,18,21,24,27,30,33-nonoxy-3,9-diazapentato Synthesis of riacontamide carbonate Step 2 of Example 15, except that compound 34-2 was replaced with compound 35-2. The title compound (20 mg) was obtained by employing a synthesis method similar to that described in ESI. -MS(m / z):1866.5[M+H].

[0257] Step 3: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2 -(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)methyl)-1 H-1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18 ,21,24,27,30,33-nonoxa-3,9-diazapentatriacontoami (D) Benzyl ((S)-4-ethyl-11-(2-(N-isopropylacetamide) (Chill)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3' ,4':6,7]Indridino[1,2-b]Quinoline-4-yl)carbonate (compound) Synthesis of material TL035) Step 3 of Example 15, except that compound 34-3 was replaced with compound 35-3. Using a synthesis method similar to the one described, the trifluoroacetate of the title compound (4. 9 mg was obtained. ESI-MS (m / z): 1594.5 [M+H] + .

[0258] Example 17: 4-((S,Z)-2-(4-aminobutyl)-42-(2-(methyls Rufonyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12,15,18 ,21,24,27,30,33-nonanoxa-3,9,36-triazadotetraconti Ru-41-Alkenamide)benzyl-((S)-4-ethyl-11-(2-(N-iso Propylacetamido(ethyl)-3,14-dioxo-3,4,12,14-tetrahydroxy Dro-1H-pyrone[3',4':6,7]indrizino[1,2-b]quinoline-4- Synthesis of yl carbonate [ka]

[0259] Step 1: (Z)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hex Synthesis of 5-enoic acid At 20°C, dissolve compounds 3-4 (200 mg, 0.67 mmol) in methanol (8.0 mL) Dissolve in ), add Lindler catalyst (20 mg) under nitrogen protection, then hydrogen purging of the solution The process was repeated three times. Hydrogenation was carried out at 20°C for 3 hours. After filtration, the filtrate was put through a dehydrator to obtain the title compound. (150 mg) was obtained. ESI-MS(m / z):271.1[M+H] + .

[0260] Step 2: (S)-4-ethyl-11-(2-(N-isopropylacetamide) Ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrone[3 ',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-((S,Z) -2-(4-(((4-methoxyphenyl)benzhydryl)amino)butyl)-42- (2-(methylsulfonyl)pyrimidine-5-yl)-4,8,37-trioxo-6, 12,15,18,21,24,27,30,33-nonanoxa-3,9,36-tria Synthesis of zadotetracontyl-41-alkeneamide)benzylcarbonate At room temperature, compound 45-2 (8 mg, 0.030 mmol) is dissolved in dichloromethane (2 mL). Dissolve in, then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyl Tyluronium hexafluorophosphate (14.9 mg, 0.039 mmol) and N,N-diisopropylethylamine (8.8 mg, 0.068 mmol) was added. The reaction solution was stirred at room temperature for 10 minutes, and then compound 48-1 (30 mg, 0.020 mm) was added. Add (ol) and react at room temperature for 1 hour with stirring. Purify by preparative high-performance liquid chromatography. The title compound (30 mg) was obtained using method B. ESI-MS (m / z): 17 87.8 [M+H] + .

[0261] Step 3: 4-((S,Z)-2-(4-aminobutyl)-42-(2-(methyl Sulfonyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12,15,1 8,21,24,27,30,33-nonanoxa-3,9,36-triazadedotetraco Nthyl-41-alkenamide)benzyl-((S)-4-ethyl-11-(2-(N- Isopropylacetamide)ethyl)-3,14-dioxo-3,4,12,14-teto Lahydro-1H-pyrone[3',4':6,7]indolidino[1,2-b]quinoline- Synthesis of 4-yl)carbonate (compound TL045) At room temperature, compound 45-3 (30 mg, 0.017 mmol) was dissolved in acetonitrile (1 ml). Dissolve in (0.5 ml) trifluoroacetic acid (0.5 ml) in acetonitrile (0.5 ml) The solution was added dropwise. The reaction solution was stirred at room temperature for 20 minutes. The purified solution was collected using preparative high-performance liquid chromatography. The trifluoroacetate salt (9 mg) of the title compound was obtained by following the Fee method (Method C). ESI- MS (m / z): 1515.6 [M+H] + .

[0262] Example 18: 4-((S)-2-(4-aminobutyl)-42-(2-(methylsulfon Nyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12,15,18,2 1,24,27,30,33-nonanoxa-3,9,36-triazadedotetracontyl -41-Alkynamide)benzyl-((S)-4-ethyl-11-(2-(N-isopropyl Ropyracetamide)ethyl)-3,14-dioxo-3,4,12,14-tetrahydr Ro-1H-pylon[3',4':6,7]indridino[1,2-b]quinoline-4-i Carbonato [ka]

[0263] Step 1: (S)-4-ethyl-11-(2-(N-isopropylacetamide) Ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrone[3 ',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-((S)-2 -(4-(((4-methoxyphenyl)benzhydryl)amino)butyl)-42-(2 -(methylsulfonyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12 ,15,18,21,24,27,30,33-nonaoxa-3,9,36-triazado Synthesis of tetracontyl-41-alkynamide)benzylcarbonate Step 1 of Example 12, except that compound 24-1 was replaced with compound 48-1. The title compound (15 mg) was obtained by employing a synthesis method similar to the one described in ESI. -MS(m / z):1785.8[M+H] + .

[0264] Step 2: 4-((S)-2-(4-aminobutyl)-42-(2-(methylsulfur Honyl)pyrimidine-5-yl)-4,8,37-trioxo-6,12,15,18, 21,24,27,30,33-nonanoxa-3,9,36-triazadedotetraconti L-41-alkynamide)benzyl-((S)-4-ethyl-11-(2-(N-iso Propylacetamido(ethyl)-3,14-dioxo-3,4,12,14-tetrahydroxy Dro-1H-pyrone[3',4':6,7]indrizino[1,2-b]quinoline-4- Synthesis of yl carbonate (compound TL048) Step 2 of Example 12, except that compound 24-2 was replaced with compound 48-2. Using a synthesis method similar to the one described, the trifluoroacetate (11 0.35 mg was obtained. ESI-MS (m / z): 1513.7 [M+H] + .

[0265] Example 19: 4-((S)-2-(4-aminobutyl)-35-(4-((2-(2- ((methylsulfonyl)pyrimidine-5-yl)thiazole-4-carboxamide) (Lu)-1H-1,2,3-triazole-1-yl)-4,8-dioxo-6,12,1 5,18,21,24,27,30,33-nonaoxa-3,9-diazapentatrico (N-isopropylmethyl)benzyl((S)-4-ethyl-11-(2-(N-isopropylmethyl) (Sulfonamide)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1 H-pyrone[3',4':6,7]indrizino[1,2-b]quinoline-4-yl) Lubonart [ka]

[0266] Step 1: 2-(2-(methylthio)pyrimidine-5-yl)thiazole-4-ca Synthesis of rubonic acid Compound 49-1 (100 mg, 0.40 mmol), 2-bromo-4-thiazole Glucoside acid (99.01 mg, 0.48 mmol), potassium carbonate (137.03 mg, 0. 99 mmol) and [1,1'-bis(diphenylphosphino)ferrocenyl]paradiul Mudicloride (29.02 mg, 0.04 mmol) is added to N,N-dimethylformamide ( Dissolve the solution in 4 mL of water (1 mL) under nitrogen protection, heat the reaction system to 100°C, and stir for 4 hours. Next, the reaction solution was cooled to room temperature and added dropwise to water. After filtration, the filtrate was collected and acetic acid was added. Extraction was performed using chill (10 mL x 3). The aqueous phase was collected and the pH was adjusted to 3 with dilute hydrochloric acid to extract the solid. The mixture was precipitated and filtered. The filtration cake was collected to obtain the title compound (70 mg). ESI-M S(m / z):254.0[M+H] + .

[0267] Step 2: 2-(2-(methylsulfonyl)pyrimidine-5-yl)thiazole- Synthesis of 4-carboxylic acids Compound 49-2 (73 mg, 0.29 mmol) was dissolved in dichloromethane (15 mL). And m-chloroperoxybenzoic acid (175.53 mg, 0.87 mmol, 85%) The solution was added. The reaction system was stirred overnight at room temperature. The solvent was concentrated under reduced pressure. The purified solution was then separated into preparative high-performance liquids. The title compound (20 mg) was obtained by chromatography (Method D). ESI-MS ( m / z): 286.0 [M+H] + .

[0268] Step 3: 2-(2-(methylsulfonyl)pyrimidine-5-yl)-N-(pro Synthesis of Pa-2-in-1-yl)thiazo-4-carboxamide Compound 49-3 (20 mg, 0.07 mmol) was dissolved in dichloromethane (2 mL). O-(7-benzotriazole)-N,N,N,N-tetramethyluronium hexaf Luorophosphate (39.98 mg, 0.11 mmol) was added. The resulting reaction system Cool to 0°C, then add N,N-diisopropylethylamine (22.65 mg, 0.1 Add 8 mmol) and propargylamine (4.63 mg, 0.09 mmol) to the reaction system. The reaction solution was stirred at room temperature for 3 hours. Purification was performed by preparative high-performance liquid chromatography ( The title compound (10 mg) was obtained by method D). ESI-MS (m / z): 323.0[ M+H] + .

[0269] Step 4: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Ron[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl-(4-( (S)-2-(4-(((4-methoxyphenyl)benzhydryl)amino)butyl)- 35-(4-((2-(2-((methylsulfonyl)pyrimidine-5-yl)thiazole- 4-Carboxamide)methyl)-1H-1,2,3-triazol-1-yl)-4,8 -Dioxo-6,12,15,18,21,24,27,30,33-Nonazeza-3,9 - Synthesis of diazapentatriaconeamino(benzyl)carbonate At room temperature, compound 33-1 (30 mg, 0.02 mmol) and compound 49-4 (9.0 Dissolve 8 mg (0.03 mmol) in dimethyl sulfoxide and water (2 mL / 0.5 mL). The mixture was dissolved, copper bromide (5.39 mg, 0.04 mmol) was added, and the mixture was reacted under stirring for 2 hours. After filtration, the filtrate is separated and purified by high-performance liquid chromatography (Method B) to obtain the title compound. (20 mg) was obtained. ESI-MS (m / z): 1647.3 [M + H - 273] + .

[0270] Step 5: 4-((S)-2-(4-aminobutyl)-35-(4-((2-(2 -((methylsulfonyl)pyrimidine-5-yl)thiazole-4-carboxamide) (Tyl)-1H-1,2,3-triazole-1-yl)-4,8-dioxo-6,12, 15,18,21,24,27,30,33-nonaoxa-3,9-diazapentatoria Contamino)benzyl((S)-4-ethyl-11-(2-(N-isopropylmethyl Sulfonamide)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro- 1H-pyrone[3',4':6,7]indrizino[1,2-b]quinoline-4-yl) Synthesis of carbonate At room temperature, compound 49-5 (20 mg, 0.01 mmol) is dissolved in dichloromethane (2 mL). The solution was dissolved in [a certain solution] and trifluoroacetic acid (0.2 mL) was added dropwise. The resulting reaction solution was heated at room temperature. The mixture was stirred for 20 minutes. The reaction solution was then concentrated. The residue was then subjected to preparative high-performance liquid chromatography. The trifluoroacetate salt (8 mg) of the title compound was purified by (Method C). ESI -MS(m / z):1647.9[M+H] + .

[0271] Example 20: 4-((S)-2-(4-aminobutyric acid)-35-(4-((2-(2-( Methylsulfonyl pyrimidine-5-yl)oxazole-4-formylamino)meth (Lu)-1H-1,2,3-triazole-1-yl)-4,8-dioxo-6,12,1 5,18,21,24,27,30,33-nonoxy-3,9-diazapentatrico (N-isopropylmethyl)benzyl-((S)-4-ethyl-11-(2-(N-isopropylmethyl Sulfonamide)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro- 1H-Pyrano[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl) Carbonato [ka]

[0272] Step 1: Ethyl 2-(2-(methylthio)pyrimidine-5-yl)oxazole Synthesis of -4-carboxylate At 25℃, ethyl 2-bromooxazole-4-carboxylate (100mg, 0. 45 mmol) and compound 49-1 (126 mg, 0.50 mmol) are 1,4-dioxy Dissolve in a mixed solvent of san and water (4 mL / 2 mL), then potassium carbonate (125 mg, 0.9 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]paradiul Mudicloride (33 mg, 0.05 mmol) was continuously added under N2 protection, and the mixture was 9 The mixture was heated to 0°C and reacted for 3 hours. The reaction solution was filtered through diatomaceous earth. The filtrate was then diluted with water. Diluted with 50 mL and extracted with ethyl acetate (30 mL x 3). Combined the organic phases and dried. The drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product. This was then separated. Purified and labeled by thin-layer chromatography (petroleum ether / ethyl acetate = 2 / 1). A compound (40 mg) was obtained. ESI-MS(m / z):266.1[M+H] + .

[0273] Step 2: 2-(2-(methylthio)pyrimidine-5-yl)oxazole-4- Synthesis of carboxylic acids Compound 50-1 (50 mg, 0.19 mmol) is dissolved in tetrahydrofuran and water at 25°C. Dissolve in a mixed solvent (4 mL / 2 mL), and after dissolution is complete, lithium hydroxide monohydrate (40 (mg, 0.94 mmol) was added to it and reacted at 25°C for 1 hour. The reaction solution was diluted with water ( The solution was diluted with 15 mL and extracted with ethyl acetate (20 mL x 2). The aqueous phase was pH-controlled with 1N dilute hydrochloric acid. Adjust the ratio to 2-3, then add the mixed solvent of dichloromethane / methanol (v:v=10:1). Extraction was performed using (20 mL x 3). The organic phase was combined and washed with saturated physiological saline (30 mL x 1). The material was then dried on anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated to obtain the standard The target compound (40 mg) was obtained. This was used directly in further reactions without purification. ES I-MS (m / z): 238.1 [M+H] + .

[0274] Step 3: 2-(2-(methylsulfonyl)pyrimidine-5-yl)oxazole Synthesis of -4-carboxylic acids Compound 50-2 (40 mg, 0.17 mmol) is dissolved in dichloromethane (6 mL) at 25°C. Dissolve in [a solution], and after dissolution is complete, m-chloroperoxybenzoic acid (29 mg, 0.17 mmol) ) was added to it, and the mixture was stirred and reacted at 25°C for 14 hours. The reaction solution was concentrated, and the residue was separated. The title compound (20 mg) was purified by high-performance liquid chromatography (Method D). Got it. ESI-MS(m / z):269.9[M+H] + .

[0275] Step 4: 2-(2-(methylsulfonyl)pyrimidine-5-yl)-N-(pro Synthesis of Pa-2-in-1-yl)-oxazole-4-formamide At 25°C, compound 50-3 (20 mg, 0.07 mmol) was dissolved in dichloromethane (4 mL). Dissolve in ) and then O-(7-azabenzotriazol-1-yl)-N,N,N',N '-Tetramethyluronium hexafluorophosphate (42 mg, 0.11 mmol) ) and N,N-diisopropylethylamine (19 mg, 0.15 mmol) administered sequentially It was added and stirred for 5 minutes. Then propargylamine (5.0 mg, 0.09 mmol) was added. The mixture was then stirred at room temperature for 30 minutes. The reaction solution was concentrated, and the residue was removed. The compound was purified by preparative high-performance liquid chromatography (Method D) to obtain the title compound (5.0 ml). g) was obtained. ESI-MS (m / z): 306.9 [M+H]+ .

[0276] Step 5: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl4-((S )-2-(4-(((4-methoxyphenyl)diphenylmethyl)amino)butyl)-3 5-(4-((2-(2-((methylsulfonyl)pyrimidine-5-yl)oxazole- 4-Formylamino)methyl)-1H-1,2,3-triazol-1-yl)-4,8 -Dioxo-6,12,15,18,21,24,27,30,33-Nonoxy-3, Synthesis of 9-diazapentatriacontoamide)benzyl)carbonate At 25℃, compound 50-4 (6.0 mg, 0.02 mmol) and compound 33-1 (3 (0 mg, 0.02 mmol) in a mixed solvent of dimethyl sulfoxide and water (2 mL / 0.5 The solution was dissolved in (mL), and copper bromide (5.0 mg, 0.04 mmol) was added in a single batch. The resulting mixture was reacted at room temperature for 2 hours. The reaction solution was filtered and preparative high-performance liquid chromatography was performed. The title compound (25 mg) was purified by Raffy (Method B). ESI-MS ( m / z): 1631.3 [(M-273+H] + .

[0277] Step 6: 4-((S)-2-(4-aminobutyric acid)-35-(4-((2-(2- (Methylsulfonyl)pyrimidine-5-yl)oxazole-4-formylamino)meth (Lu)-1H-1,2,3-triazole-1-yl)-4,8-dioxo-6,12,1 5,18,21,24,27,30,33-nonoxy-3,9-diazapentatrico (N-isopropylmethyl)benzyl-((S)-4-ethyl-11-(2-(N-isopropylmethyl Sulfonamide)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro- 1H-Pyrano[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl) Synthesis of carbonate Compound 50-5 (20 mg, 0.01 mmol) was dissolved in dichloromethane (2.0 ml) at 25°C. It was dissolved in (L). After dissolution was complete, trifluoroacetic acid (0.2 mL) was added to the reaction mixture. The reaction was carried out at 25°C for 10 minutes. The reaction solution was concentrated, and the residue was separated and subjected to high-performance liquid chromatography. The trifluoroacetate salt (3.0 mg) of the title compound was obtained by purification using method C. Ta. ESI-MS(m / z):816.5[M / 2+H] + .

[0278] Example 21: N-((1-((6S,9S)-1-amino-6-((4-(((S)- 2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)-2 -((S)-2-(dimethylamino)-3-methylbutanamide)-N,3-dimethylbutanamide Tanamid)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3- Methoxy-2-methylpropanamide)-3-phenylpropanamide)methyl)pheni Carbamoyl-9-isopropyl-1,8,11,15-tetraoxy-13,1 9,22,25,28,31,34,37,40-nononaoxa-2,7,10,16-te Traazaanthracene-42-yl)-1H-1,2,3-triazole-4-yl) (Tyl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)-5-hexinamide [ka]

[0279] Step 1: 9-Fluorenylmethyl((S)-1-(((S)-1-((4-(( (S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-(( S)-2-((S)-2-(dimethylamino)-3-butyrylamide)-N,3-Dimeth Rubyrylamide)-3-methoxy-5-methylheptanoyl)pyro-2-yl)-3- Methoxy-2-methylpropionamide)-3-phenylpropionamide)methyl) phenyl)amino)-1-oxo-5-ureidopenta-2-yl)amino)-3-methyl Synthesis of (1-oxobutan-2-yl)carbamate Compound 51-1 (100 mg, 0.17 mmol) and 9-fluorenylmethyl at room temperature ((S)-1-(((S)-1-((4-(((S)-2-amino-3-phenylpropane (amino)methyl)phenyl)amino)-1-oxo-5-pentylureido-2-yl (amino)-3-methyl-1-oxobutan-2-yl)carbamate trifluoroacetate Tart (144 mg, 0.17 mmol) in N,N-dimethylformamide (2 mL) Dissolve, cool to 0°C, then 1-hydroxybenzotriazole (34 mg, 0.25 mmol), N-methylmorpholine (51 mg, 0.51 mmol) and 1-(3-dimethylmorpholine) Tylaminopropyl)-3-ethylcarbodiimide hydrochloride (48 mg, 0.25 mmol was added continuously. After the addition, the reaction solution was stirred at 0°C for 5 hours. The sample was poured into water (20 mL) to allow the white solid to precipitate, and then filtered by suction. The resulting filter cake was then removed. The sample was washed and dried to obtain the title compound (200 mg). ESI-MS (m / z): 13 29.2[M+H] + .

[0280] Step 2: (S)-2-((S)-2-amino-3-butyrylamino)-N-(4 -(((S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4 -((S)-2-((S)-2-(dimethylamino)-3-butyrylamino)-N,3- Dimethylbutyrylamino)-3-methoxy-5-methylheptanoyl)pyro-2-yl) (-3-Methoxy-2-methylpropionamide)-3-phenylpropionamide)Meth Synthesis of phenyl-5-ureidobarrelamide At room temperature, compound 51-2 (200 mg, 0.12 mmol) was dissolved in N,N-dimethylform. The amide was dissolved in 5 mL, and piperidine (0.5 mL) was added. The reaction solution was left at room temperature for 2 minutes. After stirring for a period of time, the solution was purified by preparative high-performance liquid chromatography (Method D). The compound (65 mg) was obtained. ESI-MS(m / z):1107.2[M+H] + .

[0281] Step 3: (S)-2-((S)-35-azido-2-isopropyl-4,8-di Oxo-6,12,15,18,21,24,27,30,33-nonoxa-3,9- Diazapentatriacintoamino)-N-(4-(((S)-2-((2R,3R)-3 -((S)-1-((3R,4S,5S)-4-((S)-2-((S)-2-(Jimechi (Dimethylbutanamide)-N,3-dimethylbutanamide)-3-methylbutanamide C-5-methylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methylpro Panamide)-3-phenylpropanamide)methyl)phenyl)-5-ureidobarrel Synthesis of amides 32-Azid-5-Oxo-3,9,12,15,18,21,24,27,30-No Naoxa-6-azatricarboxylic acid (33.1 mg, 0.06 mmol) is N,N-dimethyl Dissolve in thylformamide (5 mL), then O-(7-azabenzotriazole)-N N,N,N-tetramethyluronium hexafluorophosphate (38 mg, 0.1 0 mmol) and N,N-diisopropylethylamine (26 mg, 0.20 mmol) The compound 51-3 was added. The reaction solution was stirred at room temperature for 10 minutes, then cooled to 0°C. (55 mg, 0.05 mmol) was added. The reaction solution was stirred at room temperature for 2 hours, and preparative high-speed fertilization was performed. The title compound (56 mg) was obtained by purification using liquid chromatography (Method D). ESI-MS (m / z): 821.8 [M / 2 + H] + .

[0282] Step 4: N-((1-((6S,9S)-1-amino-6-((4-(((S) -2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)- 2-((S)-2-(dimethylamino)-3-methylbutanamide)-N,3-dimethyl Butanamide)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3 -Methoxy-2-methylpropanamide)-3-phenylpropanamide)methyl)fe Nyl)carbamoyl)-9-isopropyl-1,8,11,15-tetraoxy-13, 19,22,25,28,31,34,37,40-nonanoxa-2,7,10,16- Tetraazaanthracen-42-yl)-1H-1,2,3-triazole-4-yl) Methyl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)-5-hexineamide do's synthesis At room temperature, compounds 51-4 (56 mg, 0.04 mmol) and 6-(2-(methylsulfur Honyl)pyrimidine-5-yl)-N-(propa-2-in-1-yl)-5-hexine Amid (16 mg, 0.05 mmol) is mixed with dimethyl sulfoxide and water (2 ml). It was dissolved in L / 0.5 mL and copper bromide (10 mg, 68.17 umol) was added. The mixture was stirred for 2 hours and then filtered. The filtrate was separated and subjected to high-performance liquid chromatography ( The title compound (50 mg) was purified by method D). ESI-MS (m / z): 9 74.3 [M / 2+H] + .

[0283] Example 22: 4-((2S,5S)-5-isopropyl-38-(4-((6-(2- (Methylsulfonyl)pyrimidine-5-yl)-5-hexinamide)methyl)-1H- 1,2,3-Triazole-1-yl)-4,7,11-Trioxo-2-(3-urei) Dopropyl)-9,15,18,21,24,27,30,33,36-nonoxa-3 ,6,12-Triazadotetracontyl)benzyl-((S)-1-(((S)-1-( ((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R )-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2- Methyl-3-oxypropyl)pyrrolidine-1-yl)-3-methoxy-5-methyl-1 -oxyheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2 -yl)(methyl)carbamate [ka]

[0284] Step 1: 4-((2S,5S)-38-azido-5-isopropyl-4,7,1 1-Trioxo-2-(3-ureidopropyl)-9,15,18,21,24,27, 30,33,36-nononaoxa-3,6,12-triazadotetracontyl)benzyl- ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1 R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropane-2-i (Lu)amino)-1-methoxy-2-methyl-3-oxypropyl)pyrrolidine-1-yl )-3-methoxy-5-methyl-1-oxyheptan-4-yl)(methyl)amino)- 3-Methyl-1-oxobutan-2-yl)(methyl)carbamate At room temperature, compound 53-1 (100 mg, 0.09 mmol) was dissolved in N,N-dimethylform. Dissolve in amide (3 mL), then 1-hydroxybenzotriazole (13 mg, 0. 0.9 mmol), N,N-diisopropylethylamine (36 mg, 0.28 mmol) Compound 52-1 (67 mg, 0.09 mol) was added. The reaction solution was left at room temperature for 16 hours. The mixture was stirred, then purified by preparative high-performance liquid chromatography (Method D), and finally labeled. A compound (120 mg) was obtained. ESI-MS(m / z):830.1[M / 2+H] + .

[0285] Step 2: 4-((2S,5S)-5-isopropyl-38-(4-((6-(2 -(methylsulfonyl)pyrimidine-5-yl)-5-hexinamide)methyl)-1H -1,2,3-triazole-1-yl)-4,7,11-trioxo-2-(3-ure) Idopropyl)-9,15,18,21,24,27,30,33,36-nonoxa- 3,6,12-Triazadotetracontyl)benzyl-((S)-1-(((S)-1- (((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2 R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2 -methyl-3-oxypropyl)pyrrolidine-1-yl)-3-methoxy-5-methyl- 1-Oxyheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan- Synthesis of 2-yl(methyl)carbamate At room temperature, compound 52-2 (22 mg, 0.07 mmol) was dissolved in dimethyl sulfoxide and Dissolve in a water mixture (3 mL / 0.3 mL), then add copper bromide (18 mg, 0.13 mL). (mol) was added and stirred for 1 hour. The reaction solution was filtered. The filtrate was separated and processed using high-performance liquid chromatography. The title compound (92 mg) was purified by fluoroscopy (Method D). ESI-MS ( m / z): 982.8 [M / 2 + H] + .

[0286] Example 23: (S)-2-((2R,3R)-3-((2S)-1-((3R,4S, 5S)-4-((S)-N,3-dimethyl-2-((S)-3-methyl2-((methyl(( (4-((S)-2-((S)-3-methyl-2-(32-(4-((6-(2-(Meth (Sulfonyl)pyrimidine-5-yl)hexa-5-carbamoyl)methyl)-1H-1 ,2,3-triazole-1-yl)-5-oxo-3,9,12,15,18,21, 24,27,30-nonoxy-6-azatriacontoamino)butyrylamino)-5- Ureidovalerylamino)benzyl)oxy)carbonyl)amino)butyrylamino)- 3-Methoxy-5-methylheptyl)pyrrolidine-2-yl)-3-Methoxy-2-methyl Synthesis of lupropionyl-L-phenylalanine [ka]

[0287] Step 1: 4-((2S,5S)-38-azido-5-isopropyl-4,7,1 1-Trioxo-2-(3-ureidopropyl)-9,15,18,21,24,27, 30,33,36-nonoxy-3,6,12-triazaoctatricontoamino)be Synthesis of nzyl-(4-nitrophenyl)-carbonate At 25°C, compound 29-1 (500 mg, 0.55 mmol) was converted to N,N-dimethylform Dissolve in Muamido (10 mL) and N,N-diisopropylethylamine (141 mg, 1 Add 0.09 mmol) and then di(p-nitrobenzo) in dichloromethane (1 mL). A solution of carbonate (332 mg, 1.09 mmol) was added dropwise. After addition, The mixture was reacted at 25°C for 3 hours under stirring. The reaction solution was then subjected to reverse column (C18) chromatography. The title compound (400 mg) was purified using Raffy (acetonitrile / water = 1:2). Got it. ESI-MS(m / z):1081.9[M+H] + .

[0288] Step 2: (S)-2-((2R,3R)-3-((2S)-1-((3R,4S) ,5S)-4-((S)-2-((S)-2-((((4-((S)-2-((S)-2 -(32-Azid-5-Oxo-3,9,12,15,18,21,24,27,30- Nonoxy-6-azatriacontoamide)-3-methylbutyrylamino)-5-urei Dopentanoylamino)benzyl)oxy)carbonyl)(methyl)amino)-3-meth (Rubutyrylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5-methyl Butyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropionyl)-L-Fe Nylalanine synthesis At 25℃, compound 53-1 (60 mg, 0.06 mmol) and ((2R)-3-((2 S)-1-((3R,5S)-4-((S)-N,3-dimethyl-2-((S)-3-dimethyl- Tyl-2-(methylamino)butyrylamino)butyrylamino)-3-methoxy-5-methylamino (Tylheptyl)pyrroridine-2-yl)-3-methoxy-2-methipropionyl)-L- Phenylalanine (41 mg, 0.06 mmol) is added to N,N-dimethylformamide (2 It was dissolved in (mL). After dissolution was complete, 1-hydroxybenzotriazole (8 mg, 0.0 mL) was added. 6 mmol) was added. After the addition, the mixture was stirred at 25°C for 16 hours. The reaction solution was then divided. The title compound (38 mg) was obtained by purification using high-performance liquid chromatography (Method D). ESI-MS (m / z): 837.2 [M / 2 + H] + .

[0289] Step 3: (S)-2-((2R,3R)-3-((2S)-1-((3R,4S) ,5S)-4-((S)-N,3-dimethyl-2-((S)-3-methyl2-(methyl( ((4-((S)-2-((S)-3-methyl-2-(32-(4-((6-(2-(Me (Tylsulfonyl)pyrimidine-5-yl)hexa-5-carbamoyl)methyl)-1H- 1,2,3-Triazole-1-yl)-5-oxo-3,9,12,15,18,21 ,24,27,30-nonoxy-6-azatriacontoamino)butyrylamino)-5 -Ureidovalerylamino)benzyl)oxy)carbonyl)amino)butyrylamino) -3-Methoxy-5-methylheptyl)pyrroridine-2-yl)-3-Methoxy-2-methylheptyl Synthesis of tylpropionyl-L-phenylalanine At 25℃, 2-(2-(methylsulfonyl)pyrimidine-5-yl)-N-(propa-2 -I-1-yl)-oxazole-4-formamide (9 mg, 0.03 mmol) and Compound 53-2 (50 mg, 0.03 mmol) is mixed with dimethyl sulfoxide and water (1 ml). It was dissolved in a mixed solvent of (L / 0.25 mL). After dissolution was complete, copper bromide (11 mg, 0.08 mmol was added. After addition, the mixture was stirred under N2 protection for 1 hour. Then, filtration was performed. The filtrate is purified by preparative high-performance liquid chromatography (Method D) to obtain the title compound ( 25 mg was obtained. ESI-MS (m / z): 989.9 [M / 2 + H] + .

[0290] Example 24: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2- (Methylsulfonyl)pyrimidine-5-yl)-5-hexinamide)methyl)-1H- 1,2,3-Triazol-1-yl)-4,8-Dioxo-6,12,15,18,2 1,24,27,30,33-nonoxa-3,9-diazapentatricontoamino) Benzyl-((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)- 2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropane (-2-yl)amino)-1-methoxy-2-methyl-3-oxypropyl)pyrrolidine -1-yl)-3-methoxy-5-methyl-1-oxyheptan-4-yl)(methyl) (amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-ox Sobutan-2-yl)(methyl)carbamate [ka]

[0291] Step 1: (S)-4-(35-azido-2-(4-(((4-methoxyphenyl )Benzhydryl)amino)butyryl)-4,8-dioxo-6,12,15,18,2 1,24,27,30,33-nonoxa-3,9-diazapentatricontoamino) Synthesis of benzyl-(4-nitrophenyl)-carbonate At room temperature, add 1 g (0.95 mmol) of compound 54-1 to 20 ml of dichloromethane. Dissolve, then add N,N-diisopropylethylamine (488 mg, 3.77 mmol). Add the di-(p-nitrophenyl)-carbone in dichloromethane (10 mL), followed by di-(p-nitrophenyl)-carbone in dichloromethane (10 mL). A solution of Nat (860 mg, 2.83 mmol) was added dropwise. The resulting reaction solution was then heated in a room. Stir at warm for 6 hours, then perform silica gel column chromatography (dichloromethane / methanol). The title compound (900 mg) was purified by ESI-MS (m / ). z):953.0[M+H-273] + .

[0292] Step 2: 4-((S)-35-azido-2-(4-(((4-methoxyphenyl )Benzhydryl)amino)butyryl)-4,8-dioxo-6,12,15,18,2 1,24,27,30,33-nonoxa-3,9-diazapentatricontoamino) Benzyl ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2 -((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropane -2-yl)amino)-1-methoxy-2-methyl-3-oxypropyl)pyrrolidine- 1-yl)-3-methoxy-5-methyl-1-oxyheptan-4-yl)(methyl) (amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxo Synthesis of butan-2-yl(methyl)carbamate At room temperature, dissolve 1-hydroxybenzotriazole (33 mg, 0) in compound 54-2 (2 ml). 0.25 mmol), N,N-diisopropylethylamine (48 mg, 0.37 mmol) ), then compound 52-1 (88 mg, 0.12 mmol) was added. The resulting reaction solution The solution was stirred at room temperature for 16 hours, and then subjected to preparative high-performance liquid chromatography (Method B). The compound was purified to obtain the title compound (150 mg). ESI-MS (m / z): 1803.6[ M+H] + .

[0293] Step 3: 4-((S)-2-(4-(((4-methoxyphenyl)benzhydr (L)amino)butyryl)-35-(4-((6-(2-(methylsulfonyl)pyrimidine -5-yl)-5-hexinamide)methyl)-1H-1,2,3-triazole-1- Il)-4,8-Dioxo-6,12,15,18,21,24,27,30,33-No Naoxa-3,9-diazapentatriaconeamino)benzyl((S)-1-(((S )-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-((( 1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-meth Xy-2-methyl-3-oxypropyl)pyrrolidine-1-yl)-3-methoxy-5- Methyl-1-oxyheptan-4-yl)(methyl)amino)-3-methyl-1-oxo Butan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl) Rubamart synthesis At room temperature, compound 54-3 (100 mg, 0.06 mmol) and 6-(2-(methylsulfate Rufonyl)pyrimidine-5-yl)-N-(propa-2-in-1-yl)-5-hexy Namide (26 mg, 0.08 mmol) mixed with dimethyl sulfoxide (2 mL) and water (0 Dissolve in 0.5 mL, then add copper bromide (16 mg, 0.11 mmol) and let stand for 2 hours. The mixture was stirred. Next, it was filtered, and the filtrate was separated and subjected to high-performance liquid chromatography (Method B). The compound was purified to obtain the title compound (70 mg). ESI-MS (m / z): 1936.6 [M+H-273] + .

[0294] Step 4: 4-((S)-2-(4-aminobutyl)-35-(4-((6-(2 -(methylsulfonyl)pyrimidine-5-yl)-5-hexinamide)methyl)-1H -1,2,3-triazol-1-yl)-4,8-dioxo-6,12,15,18, 21,24,27,30,33-nonoxa-3,9-diazapentatriacontoamino )benzyl-((S)-1-(((S)-1-(((3R,4S,5S)-1-((S) -2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpro Pan-2-yl)amino)-1-methoxy-2-methyl-3-oxypropyl)pyrrolidine (I-1-yl)-3-methoxy-5-methyl-1-oxyheptan-4-yl)(methyl )amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-o Synthesis of xobutan-2-yl(methyl)carbamate At room temperature, compound 54-4 (70 mg, 0.04 mmol) is dissolved in dichloromethane (2 mL). The solution was dissolved in [a certain solution] and trifluoroacetic acid (0.2 mL) was added dropwise. The resulting reaction solution was heated at room temperature. Stir for 20 minutes, then concentrate, and separate the residue by high-performance liquid chromatography (Method C). The compound was purified to obtain trifluoroacetate (55 mg) of the title compound. ESI-MS (m / z):918.8[M / 2+H] + .

[0295] Example 25: 4-((S)-2-(4-aminobutyl)-35-(4-((4-(2- (Methylsulfonyl)pyrimidine-5-yl)benzamide)methyl)-1H-1,2, 3-Triazol-1-yl)-4,8-Dioxo-6,12,15,18,21,24 ,27,30,33-nonoxy-3,9-diazapentatriacontoamide)benzyl ((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamide)ethyl )-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4 Synthesis of indridino[1,2-b]quinoline-4-yl)carbonate ':6,7] [ka]

[0296] Step 1: Methyl 4-(2-(methylthio)pyrimidine-5-yl)benzoate synthesis Compound 49-1 (252 mg, 1.0 mmol), water (3 mL), Pd (dp) at 25°C pf)Cl2 (40 mg, 0.05 mmol) and potassium carbonate (277 mg, 2.0 ml) mol) in 1,4-dioxane (5 mL) containing methyl p-bromobenzoate (215 mg, The solution was continuously added to a 1.0 mmol solution and stirred at 80°C for 4 hours. The reaction solution was then mixed with acetate. Extraction was performed using chill (30 mL x 3). The organic phase was combined, dried, and then the insoluble substances were filtered out. The residue was removed by silica gel column chromatography, and the title compound was removed. A substance (220 mg) was obtained. ESI-MS (m / z): 261.0 [M+H] + .

[0297] Step 2: Synthesis of 4-(2-(methylthio)pyrimidine-5-yl)benzoic acid At 25°C, prepare lithium hydroxide monohydrate (322 mg, 7.68 mmol) and water (3 ml ) are each compound 55-1 (500 mg, 1.92 mg) in tetrahydrofuran (3 ml). The solution was added to a solution of mmol) and stirred for 4 hours. The reaction solution was adjusted to pH 3-4 with 1N hydrochloric acid. The mixture was then extracted with ethyl acetate (20 mL x 3). The organic phases were combined and dried. Insoluble substances were removed. Remove by filtration, and purify the residue by preparative high-performance liquid chromatography (Method D). , the title compound (430 mg) was obtained. ESI-MS(m / z):246.9[M+H] + .

[0298] Step 3: Compound 4-(2-(methylsulfonyl)pyrimidine-5-yl)benzoic acid Growth At 25℃, m-chloroperoxybenzoic acid (420mg, 2.44 mmol) is dichloro Add compound 55-2 (200 mg, 0.81 mmol) to a solution in lomethane (5 ml). Then, after stirring for 5 hours, it was purified by silica gel column chromatography, and the title The compound (180 mg) was obtained. ESI-MS(m / z):279.0[M+H] + .

[0299] Step 4: 4-(2-(methylsulfonyl)pyrimidine-5-yl)-N-(pro Synthesis of per-2-in-1-yl)benzamide At 25℃, benzotriazole-N,N,N',N'-tetramethyluronium hexa Fluorophosphate (100 mg, 0.26 mmol) mixed with dichloromethane (10 mL) Add to the solution of compound 55-3 (50 mg, 0.18 mmol) and stir for 30 minutes. Next, propynylamine (10 mg, 0.2 mmol) and N,N-diisopropyl ester Add thylamine (70 mg, 0.5 mmol) to the reaction solution and stir for 2.5 hours. The reaction solution was purified by silica gel column chromatography to obtain the title compound (2 0 mg was obtained. ESI-MS (m / z): 316.0 [M+H] + .

[0300] Step 5: (S)-4-ethyl-11-(2-(N-isopropylmethanesulfone) (Amido)ethyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-p Lano[3',4':6,7]Indrizino[1,2-b]Quinoline-4-yl(4-(( S)-2-(4-(((4-methoxyphenyl)diphenylmethyl)amino)butyl) -35-(4-((4-(2-(methylsulfonyl)pyrimidine-5-yl)benzamide (D)methyl)-1H-1,2,3-triazol-1-yl)-4,8-dioxo-6, 12,15,18,21,24,27,30,33-nonoxy-3,9-diazapenta Synthesis of Triacontamide (benzyl) Carbonate Under N2 protection, at 25°C, copper iodide (10 mg, 0.05 mmol) and water (2 mL) Compound 55-4 (16 mg, 0.05 mmol) and Compound 33-1 (80 mg, 0.0 Add the solution to a 5 mmol dimethyl sulfoxide solution (2 mL) continuously and stir for 1 hour. The reaction was carried out. Purification (Method B) was performed to obtain the title compound (79 mg). ESI-MS (m / z):1641.5[M-273+H] + .

[0301] Step 6: 4-((S)-2-(4-aminobutyl)-35-(4-((4-(2 -(methylsulfonyl)pyrimidine-5-yl)benzamide)methyl)-1H-1,2 ,3-triazol-1-yl)-4,8-dioxo-6,12,15,18,21,2 4,27,30,33-nonoxy-3,9-diazapentatriacontoamide)benzyl Lu((S)-4-ethyl-11-(2-(N-isopropylmethylsulfonamide)eth (Lu)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3', Synthesis of 4':6,7] indridino[1,2-b]quinoline-4-yl)carbonate At 25°C, compound 55-5 (55 mg, 0.029 mmol) is dissolved in water / acetonitrile. Add to trifluoroacetic acid (0.5 mL) in mixed solvent (0.1 mL / 0.5 mL) and stir. The mixture was stirred and reacted for 15 minutes. The reaction solution was separated and analyzed by high-performance liquid chromatography (Method C). The trifluoroacetate salt (42 mg) of the title compound was purified using ESI-MS (m / z). ):821.0[M / 2+H] + .

[0302] Example 26: N-((1-((6S,9S)-1-amino-6-((4-((S)-3 -Azid-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-( (S)-2-((S)-2-(dimethylamino)-3-methylbutyrylamino)-N,3 (-dimethylbutyrylamino)-3-methoxy-5-methylheptyl)pyrrolidine-2-I (L)-3-methoxy-2-methylpropionamide)propyl)phenyl)carbamoyl )-9-isopropyl-1,8,11,15-tetraoxo-13,19,22,25, 28,31,34,37,40-nonoxy-2,7,10,16-tetraazadotetra Conta-42-yl)-1H-1,2,3-triazol-4-yl)methyl)-6-( 2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inylamide [ka]

[0303] Step 1: 32-(4-((6-(2-(methylsulfonyl)pyrimidine-5-i (L)Hexa-5-inamide)methyl)-1H-1,2,3-triazole-1-yl) -5-Oxo-3,9,12,15,18,21,24,27,30-Non-oxy-6- Synthesis of azadotriacontanoic acid At 20℃, compound 56-1 (750 mg, 1.28 mmol) and 6-(2-(methyl Sulfonyl pyrimidine-5-yl)-N-(propa-2-in-1-yl)hexa-5 - Inylamide (496 mg, 1.54 mmol) to dimethyl sulfoxide (10 mL) It was dissolved in [a solution] and copper bromide (465 mg, 3.21 mmol) was added in a single batch. After addition... The mixture was reacted under stirring for 12 hours. The reaction solution was filtered, and the filtrate was separated and subjected to high-performance liquid chromatography. The title compound (500 mg) was obtained by purification using Graph (Method D). ESI-M S(m / z):860.4[M+H] + .

[0304] Step 2: (9H-fluoren-9-yl)methyl((S)-1-(((S)-1 -((4-((S)-3-Azid-2-((2R,3R)-3-((S)-1-((3R ,4S,5S)-4-((S)-2-((S)-2-(dimethylamino)-3-methyl (Tyrylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5-methylhept (Pyrrolidine-2-yl)-3-methoxy-2-methylpropionamide)propyl) Phenyl)amino)-1-oxo-5-ureidopentanoylamino-2-yl)amino Synthesis of )-3-methyl-1-oxobutan-2-yl)carbamate At 25℃, (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R )-3-(((S)-1-(4-aminophenyl)-3-azidopropyl-2-yl) Mino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine-1-yl)-3 -Methoxy-5-methyl-1-oxoheptyl-4-yl)-2-((S)-2-(Dimeth (Tylamino)-3-methylbutyrylamino)-N,3-dimethylbutyrylamine(185 Dissolve mg (0.24 mmol) in N,N-dimethylformamide (5 mL), then Add HATU (137 mg, 0.36 mmol), stir for 5 minutes, then (S)- 2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amine (131 mg, 0.26 mg)-3-methylbutyrylamino)-5-ureidopentanoic acid (131 mg, 0.26 mg) (mol) was added. The mixture was stirred at room temperature for 30 minutes. The reaction solution was subjected to further reactions. Used as is. ESI-MS (m / z): 626.0 [M / 2 + H] + .

[0305] Step 3: (S)-2-((S)-2-amino-3-methylbutyrylamino)-N -(4-((S)-3-Azid-2-((2R,3R)-3-((S)-1-((3R, 4S,5S)-4-((S)-2-((S)-2-(dimethylamino)-3-methylbutyrate (Lylamino)-N,3-dimethylbutyrylamino)-3-methoxy-5-methylheptyl )pyrrolin-2-yl)-3-methoxy-2-methylpropionamide)propyl) Synthesis of henyl-5-ureidobarrelamide Add diethylamine (0.5 mL) to the reaction solution obtained in step 2 at 25°C. The reaction was allowed to proceed by stirring for 30 minutes after addition. The reaction solution was separated and subjected to high-performance liquid chromatography (Method D). The title compound (70 mg) was purified by ) and ESI-MS (m / z): 515. 0[M / 2+H] + .

[0306] Step 4: N-((1-((6S,9S)-1-amino-6-((4-((S)- 3-Azid-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4- ((S)-2-((S)-2-(dimethylamino)-3-methylbutyrylamino)-N, 3-Dimethylbutyrylamino)-3-Methoxy-5-methylheptyl)pyrrolidine-2- Iyl-3-methoxy-2-methylpropionamide)propyl)phenyl)carbamyl (Lu)-9-isopropyl-1,8,11,15-tetraoxo-13,19,22,25 ,28,31,34,37,40-nonoxy-2,7,10,16-tetraazadotheto Laconta-42-yl)-1H-1,2,3-triazole-4-yl)methyl)-6- Synthesis of (2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inylamide At 25℃, (S)-2-((S)-2-amino-3-methylbutyrylamino)-N-( 4-((S)-3-Azid-2-((2R,3R)-3-((S)-1-((3R,4S ,5S)-4-((S)-2-((S)-2-(dimethylamino)-3-methylbutyryl (amino)-N,3-dimethylbutyrylamino)-3-methoxy-5-methylheptyl) (Loridine-2-yl)-3-methoxy-2-methylpropionamide)propyl)phen (Lu)-5-Ureidobarrelamide (95 mg, 0.092 mmol) and 32-(4-( (6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide) (Til)-1H-1,2,3-triazole-1-yl)-5-oxo-3,9,12,1 5,18,21,24,27,30-nonoxy-6-azadotricontanoic acid (79m Dissolve g (0.092 mmol) in N,N-dimethylformamide (4 mL) and HAT U (70 mg, 0.184 mmol) was added in a single batch. The mixture was left at room temperature for 1 hour. The mixture was stirred. The reaction solution was purified by preparative high-performance liquid chromatography (Method D) and the title was obtained. The compound (30 mg) was obtained. ESI-MS (m / z): 935.8 [M / 2 + H] + .

[0307] III. Coupling of compounds containing bioactive molecules and linkers with antibodies Example 27: Preparation of BT001002 0.3 mL of the antibody sacituzumab (anti-Trop-2, 33.5 mg / mL) was administered at 20 mM. A solution containing PB, 150 mM NaCl, and 20 mM sodium edetate (pH 7). 6) Dilute with 0.25 ml, and add 20 mM PB and 150 mM NaCl. 0.45 ml of the solution (pH 7.6) was added and mixed uniformly. The mixture was then mixed with 1 M K2HP. Adjust the pH to 7.4 with O4 solution, then add 10 mM TCEP (Tris(2-carboxymethylcellulose)). A thio(phospho) solution was added and mixed uniformly, then left at room temperature for 30 minutes. Add 15 equivalents of TL003 dissolved in dimethyl sulfoxide to the liquid system and mix uniformly. The mixture was combined and left at room temperature for 2 hours. After addition, 6.1 μl of 100 mM cysteine ​​was added and the mixture was turned over. The response was stopped. Finally, the buffer was transferred to a G-25 gel column at pH 6.44 for 20 minutes. Replace with M PB buffer solution to generate coupling between TL003 and sacituzumab. I obtained the object and named it BT001002. [ka]

[0308] Example 28: Preparation of BT001004 Sacituzumab 0.285 mL (anti-Trop-2, 17.6 mg / mL) diluted with 0.0 mg / mL of diluent. 95 mL (20 mM PB, 150 mM NaCl and 20 mM sodium edetate, p The solution was diluted with a solution containing H7.6. Then, the diluted solution was treated with a 1M Na2HPO4 solution. Adjust the pH to 7.4, add 10 mM TCEP solution, mix uniformly, and leave at room temperature for 30 minutes. It was left standing. To this solution system, 9 equivalents of TL019 dissolved in dimethyl sulfoxide were added. Add the mixture, mix thoroughly, and leave at room temperature for 2 hours. Finally, transfer the buffer to a G-25 gel column. Replace the PBS buffer solution with pH 6.5, and add TL019 and sacituzumab. We obtained a coupling product, which we named BT001004. [ka]

[0309] Example 29: Preparation of BT001012 Example 27, except that TL003 was replaced with the trifluoroacetate of TL024. Using a method similar to the one described in [reference], coupling generation of TL024 and sacituzumab I obtained the object and named it BT001012. [ka]

[0310] Example 30: Preparation of BT001013 Example 27, except that TL003 was replaced with the trifluoroacetate of TL048. Using a method similar to the one described in [reference], coupling generation of TL048 and sacituzumab I obtained the object and named it BT001013. [ka]

[0311] Example 31: Preparation of BT001018 The method is the same as the method in Example 27, except that TL003 was replaced with TL030. By employing this method, a coupling product of TL030 and sacituzumab was obtained, and this was then used in BT0 I named it 01018. [ka]

[0312] Example 32: Preparation of BT001021 Sacituzumab 0.3 mL (anti-Trop-2, 33.5 mg / mL), 20 mM PB A solution containing 150 mM NaCl and 20 mM sodium edetate (pH 7.6) Dilute with 0.25 ml, then add a solution containing 20 mM PB and 150 mM NaCl. 0.45 mL of solution (pH 7.6) was added and mixed uniformly. The mixture was then combined with 1 M Na2HPO4. Adjust the pH to 7.4 with solution 4, then add 10 mM TCEP(tris(2-carboxy)). Ethyl phosphine solution was added and mixed uniformly, then left at room temperature for 30 minutes. Add 10 equivalents of TL033 trifluoroacetate dissolved in dimethyl sulfoxide to the system. It was added, mixed uniformly, and left at room temperature for 2 hours. Then, 100 mM cysteine ​​6.1 The reaction was stopped by adding μl of buffer. Finally, the buffer was passed through a G-25 gel column. Replace with H6.5 PBS buffer solution and couple TL033 with sacituzumab. We obtained a compound product and named it BT001021. [ka]

[0313] Example 33: Preparation of BT001022 Example 27, except that TL003 was replaced with the trifluoroacetate of TL034. Using a method similar to the one described in [reference], coupling generation of TL034 and sacituzumab I obtained the object and named it BT001022. [ka]

[0314] Example 34: Preparation of BT001023 Example 27, except that TL003 was replaced with the trifluoroacetate of TL035. Using a method similar to the one described in [reference], coupling generation of TL035 and sacituzumab I obtained the object and named it BT001023. [ka]

[0315] Example 35: Preparation of BT001032 Example 27, except that TL003 was replaced with the trifluoroacetate of TL045. Using a method similar to the one described in [reference], coupling generation of TL045 and sacituzumab I obtained the object and named it BT001032. [ka]

[0316] Example 36: Preparation of BT001033 Replace TL003 with the trifluoroacetate of TL033, and replace sacituzumab with antibody M1. Except for the substitution, the same method as described in Example 27 was employed to produce TL033 We obtained a coupling product with antibody M1, which we named BT001033. [ka]

[0317] Example 37: Preparation of BT001034 Replace TL003 with the trifluoroacetate of TL033, and replace sacituzumab with antibody M2. Except for the substitution, the same method as described in Example 27 was employed to produce TL033 We obtained a coupling product with antibody M2, which we named BT001034. [ka]

[0318] Example 38: Preparation of BT001035 Antibody M3 0.3mL (anti-Trop-2, 33.5mg / mL) in 20mM PB, 1 A solution containing 50 mM NaCl and 20 mM sodium edetate (pH 7.6) 0. Dilute with 25 ml, then prepare a solution containing 20 mM PB and 150 mM NaCl ( (pH 7.6) 0.45 mL was added and mixed uniformly. The mixture was dissolved in 1 M Na2HPO4. Adjust the pH to 7.4 with the solution, then add 10 mM TCEP(tris(2-carboxyethyl acetate)). Add the phosphine solution and mix uniformly, then leave at room temperature for 30 minutes. Add 10 equivalents of TL033 trifluoroacetate dissolved in methyl sulfoxide. The mixture was then uniformly mixed. The resulting mixture was left at room temperature for 2 hours. Then, a 100 mM system solution was added. The reaction was stopped by adding 6.1 μl of phosphate solution. Finally, the buffer was transferred to a G-25 gel column. Replace with PBS buffer solution at pH 6.5, and then combine TL033 and antibody M3. We obtained a plucking product, which we named BT001035. [ka]

[0319] Example 39: Preparation of BT001036 Replace TL003 with the trifluoroacetate of TL033, and replace sacituzumab with trastuzumab Except for replacing mab, the same method as described in Example 27 was employed, We obtained a coupling product of 033 and trastuzumab, and named it BT001036. Ta. [ka]

[0320] Example 40: Preparation of BT001040 Example 27, except that TL003 was replaced with the trifluoroacetate of TL049. Using a method similar to the one described in [reference], coupling generation of TL049 and sacituzumab I obtained the object and named it BT001040. [ka]

[0321] Example 41: Preparation of BT001041 Example 27, except that TL003 was replaced with the trifluoroacetate of TL050. Using a method similar to the one described in [reference], coupling between TL050 and sacituzumab is generated. I obtained the object and named it BT001041. [ka]

[0322] Example 42: Preparation of BT001042 The method is the same as the method in Example 27, except that TL003 was replaced with TL051. By employing this method, a coupling product of TL051 and sacituzumab was obtained, and this was converted to BT0 I named it 01042. [ka]

[0323] Example 43: Preparation of BT001043 The method is the same as the method in Example 27, except that TL003 was replaced with TL052. By employing this method, a coupling product of TL052 and sacituzumab was obtained, and this was converted to BT0 I named it 01043. [ka]

[0324] Example 44: Preparation of BT001044 The method is the same as the method in Example 27, except that TL003 was replaced with TL053. By employing this method, a coupling product of TL053 and sacituzumab was obtained, and this was converted to BT0 I named it 01044. [ka]

[0325] Example 45: Preparation of BT001045 Example 27, except that TL003 was replaced with the trifluoroacetate of TL054. Using a method similar to the one described in [reference], TL054 was coupled with sacituzumab to generate [a specific product]. I obtained the object and named it BT001045. [ka]

[0326] Example 46: Preparation of BT001046 Example 27, except that TL003 was replaced with the trifluoroacetate of TL055. Using a method similar to the one described in [reference], coupling generation of TL055 and sacituzumab I obtained the object and named it BT001046. [ka]

[0327] Example 47: Preparation of BT001047 The method is the same as the method in Example 27, except that TL003 was replaced with TL056. By employing this method, a coupling product of TL056 and sacituzumab was obtained, and this was converted to BT0 I named it 01047. [ka]

[0328] Example 48: Determination of the molecular weight of BT001002 by LC-MS The molecular weight of BT001002 obtained by coupling was analyzed by LC-MS. Ta. LC conditions: Liquid chromatography column: ACQUITU UPLC(registered trademark) Prot ein BEH C4 1.7μm,2.1mm×100mm; Mobile phase A: 0.1%FA / 98%H2O / 2%ACN; Mobile phase B: 0.1%FA / 2 %H2O / 98%ACN; Flow rate: 0.25 mL / min; Sample chamber temperature: 8°C; Column temperature: 60°C; Sample Iz: 1 μg; [Table 7] MS conditions: Mass spectrometer model: Triple TOF 5600+; GS1 60;GS2 60;CUR30;TEM600;ISVF5000;DP 300;CE10 m / z600-5000; The results are shown in Figures 1-3.

[0329] Theoretical molecular weight and measured molecular weight of BT001002 [Table 8]

[0330] In the table, mAb represents a monoclonal antibody; LC represents the light chain of the antibody; and HC represents the antibody. Represents the heavy chain of the body; DAR1 contains one antibody light / heavy chain and one bioactive molecule. DAR2 represents a conjugate consisting of one antibody light / heavy chain and two bioactive molecules. DAR3 represents a conjugate containing one antibody's light / heavy chain and three organisms. DAR4 represents a conjugate containing an active molecule; DAR4 is the light / heavy chain of one antibody and 4 The term "conjugate" refers to a conjugate containing two bioactive molecules; the "glyco" type consists of two heavy chains. The structure of lycan is represented: G0F represents fucosylation lacking galactosylation. The following mAb, LC, HC, DAR1, DAR2, DAR3, DAR4, and G0 are used in this context. F is as described above.

[0331] As can be seen from Figures 1-3, the molecular weights of both the light and heavy chains of the antibody are TL003 and K After coupling, it changes, where the light chain is coupled with one bioactive molecule. The heavy chain was coupled with three bioactive molecules. Therefore, it provides resistance to the bioactive molecules. It can be inferred that the body's DAR was 8.

[0332] Example 49: Determination of the molecular weight of BT001004 by LC-MS The molecular weight of BT001004 obtained by coupling was analyzed by LC-MS. Ta. LC conditions: Liquid chromatography column: ACQUITU UPLC(registered trademark) Prot ein BEH C18 1.7μm,2.1mm×100mm; Mobile phase A: 0.1%FA / 98%H2O / 2%ACN; Mobile phase B: 0.1%FA / 2 %H2O / 98%ACN; Flow rate: 0.25 mL / min; Sample chamber temperature: 8°C; Column temperature: 60°C; Sample Iz: 1 μg; [Table 9] MS conditions: Mass spectrometer model: Triple TOF 5600+; GS1 60;GS2 60;CUR30;TEM 350;ISVF5500;D P300;CE10;m / z600~5000; The results are shown in Figures 4-6.

[0333] Theoretical molecular weight and measured molecular weight of BT001004 [Table 10] LC represents the light chain of the antibody; HC represents the heavy chain of the antibody.

[0334] As can be seen from Figures 4-6, in BT001004, the antibody light chain has 0-1 biological activity. It was coupled with molecules (LC and DAR1 accounted for 14% and 86%, respectively), and The chain was coupled with 1 to 3 bioactive molecules (DAR1, DAR2 and DAR3 are (These accounted for 13%, 19%, and 68%, respectively.) Therefore, antibodies against biologically active molecules The DAR can be calculated as 7.0.

[0335] Example 50: Determination of the molecular weight of BT001012 by LC-MS The same method as described in Example 48 was employed, and the results are shown in Figures 10 and 11.

[0336] Light chain of BT001012 obtained by coupling TL024 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated from the main glyco-type G0Fs) are shown in the table below. . [Table 11]

[0337] As can be seen from Figures 10 and 11, in BT001012, the antibody light chain has 0 to 1 toxin. The element was coupled with (LC and DAR1 accounted for 12.9% and 87.1%, respectively). ), the heavy chain was coupled with 1 to 3 toxins (DAR1, DAR2 and DAR3 are... (These accounted for 3.4%, 10.8%, and 75.8%, respectively.) Therefore, antibodies against toxins The DAR can be calculated as 7.0.

[0338] Example 51: Determination of the molecular weight of BT001013 by LC-MS The same method as described in Example 48 was employed, and the results are shown in Figures 12 and 13.

[0339] Light chain of BT001013 obtained by coupling TL048 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 12]

[0340] As can be seen from Figures 12 and 13, in BT001013, the antibody light chain has 0 to 1 toxin. Coupled with the original (LC and DAR1 accounted for 6.8% and 93.2%, respectively) The heavy chain was coupled with 1 to 4 toxins (DAR1, DAR2, DAR3 and DA R4 accounted for 12.8%, 12.8%, 64.9%, and 9.5%, respectively. Therefore, the DAR of the antibody against the toxin can be calculated to be 7.3.

[0341] Example 52: Determination of the molecular weight of BT001018 by LC-MS The same method as described in Example 48 was employed, and the results are shown in Figures 14 and 15.

[0342] Light chain of BT001018 obtained by coupling TL030 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 13]

[0343] As can be seen from Figures 14 and 15, in BT001018, the antibody light chain has 0 to 1 toxin. The elements were coupled together (LC and DAR1 accounted for 55.3% and 44.7%, respectively). The heavy chain was coupled with 1 to 3 toxins (DAR1, DAR2, DAR3 and D AR4 accounted for 19.6%, 23.3%, and 49.6%, respectively. Therefore, toxins The DAR of the antibody against can be calculated as 5.2.

[0344] Example 53: Determination of the molecular weight of BT001021 by LC-MS The molecular weight of the coupled BT001021 was analyzed by LC-MS. LC conditions: Liquid chromatography column: ACQUITU UPLC(registered trademark) Prot ein BEH C4 1.7μm,2.1mm×100mm; Mobile phase A: 0.1%FA / 98%H2O / 2%ACN; Mobile phase B: 0.1%FA / 2 %H2O / 98%ACN; Flow rate: 0.25 mL / min; Sample chamber temperature: 8°C; Column temperature: 60°C; Sample Iz: 1 μg; [Table 14] MS conditions: Mass spectrometer model: Triple TOF 5600+; GS1 60;GS2 60;CUR30;TEM600;ISVF5000;DP 300;CE10 m / z600-5000; The results are shown in Figures 16 and 17.

[0345] Light chain of BT001021 obtained by coupling TL033 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 15]

[0346] As can be seen from Figures 16 and 17, in BT001021, the antibody light chain has 0 to 1 toxin. Coupled with the original (LC and DAR1 accounted for 4.5% and 95.5%, respectively) The heavy chain was coupled with 1 to 3 toxins (DAR1, DAR2, DAR3 and DA R4 accounted for 15.3%, 17.6%, and 67.1%, respectively. Therefore, the toxins The DAR of the antibody can be calculated to be 6.9.

[0347] Example 54: Determination of the molecular weight of BT001023 by LC-MS The same method as described in Example 48 was employed, and the results are shown in Figures 18 and 19.

[0348] Light chain of BT001023 obtained by coupling TL035 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 16]

[0349] As can be seen from Figures 18 and 19, in BT001023, the antibody light chain has 0 to 1 toxin. It was coupled with the element (LC and DAR1 accounted for 15% and 85% respectively), and the heavy chain It was coupled with 0-3 toxins (HC, DAR1, DAR2 and DAR3 are...) (These accounted for 6.7%, 16.7%, 12.7%, and 63.9%, respectively.) Therefore, the toxins The DAR of the antibody can be calculated to be 6.4.

[0350] Example 55: Determination of the molecular weight of BT001040 by LC-MS The molecular weight of BT001040 obtained by coupling was analyzed by LC-MS. Ta. Liquid chromatography column: Thermo MabPac™ RP 4μm 3.0mm x 100mm Mobile phase A: 0.1%FA / 98%H2O / 2%ACN; Mobile phase B: 0.1%FA / 2 %H2O / 98%ACN Flow rate: 0.25 mL / min; Sample chamber temperature: 8°C; Column temperature: 60°C; Sample Iz: 1 μg [Table 17] MS conditions: Mass spectrometer model: Triple TOF 5600+ GS1 35;GS2 35;CUR30;TEM350;ISVF5000;DP 250; m / z 600~5000

[0351] Light chain of BT001040 obtained by coupling TL049 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 18]

[0352] As can be seen from Figures 20 and 21, in BT001040, the antibody light chain has 0 to 1 living It is coupled with the phytoactive molecule (LC and DAR1 make up 4.9% and 95.1%, respectively). The heavy chains were coupled with 1-4 bioactive molecules (DAR1, DAR2, D AR3 and DAR4 accounted for 16.5%, 14.3%, 52.6%, and 16.6%, respectively. Therefore, the DAR of the antibody against the biologically active molecule can be calculated as 7.3.

[0353] Example 56: Determination of the molecular weight of BT001041 by LC-MS The same method as described in Example 55 was employed, and the results are shown in Figures 22 and 23.

[0354] Light chain of BT001041 obtained by coupling TL050 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 19]

[0355] As can be seen from Figures 22 and 23, in BT001041, the antibody light chain has 0 to 1 living light chain. Coupled with the phytoactive molecule (LC and DAR1 are 10.5% and 89.5%, respectively) The heavy chain (occupied) was coupled with 1 to 4 bioactive molecules (DAR1, DAR2) DAR3 and DAR4 were 21.3%, 14.8%, 57.9%, and 6.0%, respectively. (Occupied). Therefore, the DAR of the antibody against the biologically active molecule can be calculated as 6.8.

[0356] Example 57: Determination of the molecular weight of BT001042 by LC-MS The same method as described in Example 55 was employed, and the results are shown in Figures 24 and 25.

[0357] Light chain of BT001042 obtained by coupling TL051 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 20]

[0358] As can be seen from Figures 24 and 25, in BT001042, the antibody light chain has 0 to 1 living Coupled with the phytoactive molecule (LC and DAR1 were 14.9% and 85.1%, respectively) The heavy chain (which occupied a portion of the total) was coupled with 1 to 3 bioactive molecules (DAR1, DAR2) (And DAR3 accounted for 19.7%, 9.4%, and 70.9%, respectively). Therefore, The DAR of an antibody against a biologically active molecule can be calculated to be 6.7.

[0359] Example 58: Determination of the molecular weight of BT001043 by LC-MS The same method as described in Example 55 was employed, and the results are shown in Figures 26 and 27.

[0360] Light chain of BT001043 obtained by coupling TL052 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 21]

[0361] As can be seen from Figures 26 and 27, in BT001043, the antibody light chain has 0 to 1 living It is coupled with the phytoactive molecule (LC and DAR1 make up 9.1% and 90.9%, respectively). The heavy chain was coupled with 1 to 3 bioactive molecules (DAR1, DAR2, and (B and DAR3 accounted for 20.1%, 11.4%, and 68.4%, respectively). Therefore, The DAR of an antibody against a biologically active molecule can be calculated to be 6.8.

[0362] Example 59: Determination of the molecular weight of BT001044 by LC-MS The same method as described in Example 55 was employed, and the results are shown in Figures 28 and 29.

[0363] Light chain of BT001044 obtained by coupling TL053 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 22]

[0364] As can be seen from Figures 28 and 29, in BT001044, the antibody light chain has 0 to 1 living light chain. Coupled with the phytoactive molecule (LC and DAR1 are 23.0% and 77.0%, respectively) The heavy chain (which occupied a portion of the total) was coupled with 1 to 3 bioactive molecules (DAR1, DAR2) (And DAR3 accounted for 19.4%, 11.4%, and 69.3%, respectively). Therefore, the DAR of the antibody against the biologically active molecule can be calculated to be 6.5.

[0365] Example 60: Determination of the molecular weight of BT001046 by LC-MS The same method as described in Example 55 was employed, and the results are shown in Figures 30 and 31.

[0366] Light chain of BT001046 obtained by coupling TL055 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 23]

[0367] As can be seen from Figures 30 and 31, in BT001046, the antibody light chain has 0 to 1 living light chain. Coupled with the phytoactive molecule (LC and DAR1 accounted for 33.8% and 66.2%, respectively) The heavy chains (which occupied a large portion of the total) were coupled with 0-3 bioactive molecules (DAR0, DAR1) DAR2 and DAR3 accounted for 21.9%, 6.1%, 9.6%, and 62.3%, respectively. Therefore, the DAR of the antibody against the biologically active molecule can be calculated as 5.6.

[0368] Example 61: Determination of the molecular weight of BT001047 by LC-MS The same method as described in Example 55 was employed, and the results are shown in Figures 32 and 33.

[0369] Light chain of BT001047 obtained by coupling TL056 and antibody. The theoretical and measured molecular weights of the heavy chains (calculated based on the main glyco-type G0F) are shown in the table below. did. [Table 24]

[0370] As can be seen from Figures 32 and 33, in BT001047, the antibody light chain has 0 to 1 living light chain. Coupled with the phytoactive molecule (LC and DAR1 were 13.7% and 86.3%, respectively) The heavy chain (which occupied a portion of the total) was coupled with 1 to 3 bioactive molecules (DAR1, DAR2) (And DAR3 accounted for 22.2%, 13.5%, and 64.3%, respectively). Therefore, the DAR of the antibody against the biologically active molecule can be calculated to be 6.6.

[0371] Example 62: Size exclusion chromatography analysis The coupling reaction is monitored by SEC-HPLC, and the conjugate is converted to SEC. Therefore, the test was conducted. Chromatography conditions: Liquid chromatography column: TOSOH TSKgel SuperSW m Ab, 4μm, 7.8mm × 300mm; Mobile phase: 100mmol / L Na2HPO4,100mmol / L NaCl,5 % isopropanol, pH 7.0; Flow rate: 0.5 ml / min; Detection wavelength: 280 nm; Column temperature: Room temperature; Sample room temperature :8℃; Sample size: 30 μg; Calculation time: 30 minutes.

[0372] SEC chromatogram of BT001002 obtained by coupling TL003 with antibody The molecular weight chromatogram and molecular weight marker SEC chromatogram are shown in Figures 7 and 8, respectively. According to the quantitative marker, the molecular weight of the main peak of the coupling product is approximately 150 kD. In other words, in BT001002 obtained by coupling TL003 with an antibody, It was confirmed that the light and heavy chains had not dissociated, and the antibody still maintained its complete structure. It can be done.

[0373] SEC chromatogram of BT001004 obtained by coupling TL019 with antibody The togram is shown in Figure 9. According to the retention time and peak area ratio in SEC, the main cup The molecular weight of the ring product is approximately 150 kD, i.e., the coupling between TL019 and the antibody. The BT001004 obtained by the process still maintains the complete structure of the antibody. It is recognized.

[0374] SEC chromatogram of BT001012 obtained by coupling TL024 with antibody The togram is shown in Figure 34. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL024 and the antibody. The BT001012 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0375] SEC chromatogram of BT001013 obtained by coupling TL048 with antibody The togram is shown in Figure 35. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL048 and the antibody. The BT001013 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0376] SEC chromatogram of BT001018 obtained by coupling TL030 with antibody The togram is shown in Figure 36. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL030 and the antibody. The BT001018 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0377] SEC chromatogram of BT001021 obtained by coupling TL033 with antibody The togram is shown in Figure 37. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL033 and the antibody. The BT001021 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0378] SEC chromatogram of BT001023 obtained by coupling TL035 with antibody The togram is shown in Figure 38. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL035 and the antibody. The BT001023 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0379] SEC chromatogram of BT001042 obtained by coupling TL051 with antibody The togram is shown in Figure 39. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL051 and the antibody. The BT001042 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0380] SEC chromatogram of BT001043 obtained by coupling TL052 with antibody The togram is shown in Figure 40. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL052 and the antibody. The BT001043 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0381] SEC chromatogram of BT001044 obtained by coupling TL053 with antibody The togram is shown in Figure 41. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL053 and the antibody. The BT001044 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0382] SEC chromatogram of BT001046 obtained by coupling TL055 with antibody The togram is shown in Figure 42. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL055 and the antibody. The BT001046 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0383] SEC chromatogram of BT001047 obtained by coupling TL056 with antibody The togram is shown in Figure 43. According to the retention time and peak area ratio in SEC, the main cuts The molecular weight of the pulling product is approximately 150 kD, i.e., the coupling between TL056 and the antibody. The BT001047 obtained by the ring still maintains the complete structure of the antibody. Confirmed.

[0384] Example 63: Bioactive molecules and antibody-drug conjugates for in vitro cell activity Testing of the inhibitory effect of t First, tumor cells MDA-MB-468 (Trop-2 positive cell line) and HCC180 Cell 6 (Trop-2 positive cell line) was cultured. The bioactive molecules and ADC molecules disclosed herein. The tumor cells were co-cultured with the CCK8 reagent (Dojindo Molecula). r Technologies, Inc., Cat: CK04, Lot: JJ744) Added. Mitochondrial dehydrogenase activity was measured using a microplate reader. (Manufacturer: Molecular Devices, model: SpectraMax The test was conducted using the reading from M2 (the detection wavelength was 450 nm) to determine the effect on cell proliferation. The inhibitory effect of ADCs was evaluated. The sources of tumor cells are shown in Table 1. [Table 25]

[0385] In vitro cell activity testing: Bioactive molecules or ADCs are tested in the corresponding test medium (2% FBS). Diluted with (12 concentration gradients). Tumor cells were treated with trypsin by conventional methods. The samples were psin-treated, collected, counted, and then re-cured in the corresponding test medium (containing 2% FBS). The solution was turbid. Diluted bioactive molecules or ADCs were added to a 96-well plate, and then the cells were... Added. Add 20 μL of CCK8 reagent to each well and allow to react for 4 hours, then measure the reading (detection wavelength). The wavelength (450 nm) was obtained from a microplate reader. Experimental conditions and test results This is shown in Tables 2 and 3.

[0386] [Table 26]

[0387] The test results showed that all of the bioactive molecules had a cell-killing effect against tumor cells. .

[0388] [Table 27]

[0389] The test results showed that ADC molecules obtained by the novel coupling method kill tumor cells. This demonstrated that it had a decompression effect. This is the ADC formed by the novel coupling method. It has a death effect on tumor cells, and a novel coupling method is used in the synthesis of ADC molecules. This indicates that it was feasible.

[0390] Example 64 Pharmacodynamic studies of antibody-drug conjugates and bioactive molecules in vivo Test drug Drug name, source, and preparation method: BT001021, liquid alicot is stored at a concentration of 5.44 mg / ml at -20°C. The test solution was prepared by diluting it to the appropriate dose with physiological saline before use. Immu-132 (prepared according to Example 2 of International Publication No. 2015 / 012904A2) (Also described as DAR=5.4, IMMU-132), liquid alicot at -20°C for 1 Store at a concentration of 3.158 mg / ml, and dilute with physiological saline to the appropriate dose before use to create the test solution. Obtained; The solid powder of T-030 was diluted with 100% DMSO (Sigma) to a concentration of 5.2 mg / mL. Prepare the solution, store the liquid alicot at -20°C, and dilute with physiological saline to the desired volume before use. The test solution was obtained by dilution; Solid powder of SN-38 (also written as SN38) is mixed with 100% DMSO (Sigma). Prepare a solution with a concentration of 3.23 mg / ml, store the liquid alicot at -20°C, and use it. The test solution was previously obtained by diluting it to the prescribed dose with physiological saline. Note: The toxin was prepared and administered in equimolar ratios to the ADC sample.

[0391] The structures of T-030, SN-38, and Immu-132 were as follows: [ka]

[0392] Experimental animals and cell lines Balb / c-nu mouse (Beijing Vital River Labora Tory Animal Technology Co., Ltd., Production License Number: SCXK (Beijing) 2016-0011); Gastric cancer cell line NCI-N87 (AT CC), breast cancer cell line HCC1806 (COBIOER Nanjing).

[0393] Experimental grouping and evaluation methods 100-200mm 3 Tumor-bearing mice (6 mice / group) with a tumor volume were randomly assigned to a group. The number of groups was determined according to the sample size. The dose volume was 10 ml / kg. The administration route was intravein injection. Mice were given the drug twice a week, and the tumor diameter was measured with calipers. The ulcer volume was calculated based on the following formula: V = 0.5a × b 2 [In the formula, a and b are respectively This represents the longest and shortest diameters of the tumor. Animal deaths were observed and recorded daily.

[0394] The tumor growth inhibition rate (TGI) (%) is calculated using the following formula to determine the tumor inhibition of the antibody-drug conjugate. The harmful effects were evaluated. TGI(%)=[1-(V Tend -V Tstart ) / (V Cend -V Csta rt )] × 100% [In the formula, V Tend : Average tumor volume at the end of the experiment in the treatment group V Tstart : Mean tumor volume at the start of administration in the treatment group V Cend : Average tumor volume at the end of the experiment in the control group V Cstart [Mean tumor volume at the start of treatment in the control group]

[0395] In the following experimental examples 1 and 2, tumor-bearing tumors were constructed using subcutaneous xenografts of human tumor cells. The inhibition of tumor growth in Ussica albicans by the antibody conjugate BT001021 was evaluated. In experimental examples 1 and 2, the tumor-bearing mouse model was the human gastric cancer cell line NCI-N87 or Constructed using subcutaneous xenografts of the human triple-negative breast cancer cell line HCC1806. Tumor volume Approximately 100mm3 After that, the mice were randomly grouped and given BT001021 twice a week. The drug was administered intravenously a total of six times. Changes in tumor volume and animal body weight were measured twice a week. The efficacy (tumor inhibitory effect) of antibody-drug conjugates against ulcerated mice was evaluated.

[0396] Experimental Example 1 Inhibition of NCI-N87 by antibody-drug conjugates and bioactive molecules. Experimental method: NCI-N87 cells were cultured in 1640 culture medium containing 10% fetal bovine serum at 37°C and NCI-N87 cells were cultured in 5% CO2. Exponential growth phase NCI-N87 cells were collected and properly cultured in PBS. The solution was resuspended to the correct concentration and subcutaneously inoculated into female Balb / c-nu mice to construct a gastric cancer model. The average tumor volume was approximately 90 mm. 3 When this happens, mice are given physiological salt according to tumor size. Water group, BT001021 (3 mg / kg, IV, BIW x 3W) group, positive drug immunoassay -132 (3 mg / kg, IV, BIW × 3W) group, T030 group, and SN38 group were randomly assigned. The subjects were grouped together, and subsequently, the corresponding drug was administered via tail vein injection twice a week for a total of six times. Subsequently, the tumor volume and body weight of the mice were observed and measured periodically. Specific results are shown in Table 4 and Figure 4. This is shown in 4 and 45.

[0397] Conclusion: In the experimental example, the human gastric cancer cell line NCI-N87 was used to study subcutaneous xenografts of human gastric cancer. Dell constructed BT00102 in a mouse model of NCI-N87 human gastric cancer. The effectiveness of option 1 was evaluated.

[0398] The experimental results showed that BT001021 (3 mg / kg, IV, BIW × 3W) was NCI-N8 7. Significantly inhibited tumor growth in xenograft model mice for gastric cancer, and tumor regression occurred at the end of administration. It was shown that shrinkage can occur, and the antitumor activity was greater than that of the positive control Immu-132. It was excellent. During the observation period, in all treatment groups, there was no significant animal death or significant animal weight loss. No adverse reactions occurred, indicating that BT001021 did not possess significant toxicity.

[0399] [Table 28]

[0400] Experimental Example 2 Inhibition of HCC1806 by antibody-drug conjugates. Experimental method: HCC1806 cells were cultured in 1640 culture medium containing 10% fetal bovine serum at 37°C. HCC1806 cells were cultured in 5% CO2. Exponential growth phase HCC1806 cells were collected and properly incubated in PBS. The solution was resuspended at the correct concentration and subcutaneously inoculated into female Balb / c-nu mice to construct a breast cancer model. The average tumor volume is approximately 130 mm. 3 When this happens, mice are given physiological salt according to tumor size. Water group, BT001021 (10 mg / kg, IV, BIW × 3W) group and positive drug Im Students were randomly assigned to the mu-132 (10 mg / kg, IV, BIW × 3W) group, followed by the opposing group. The corresponding drug was administered via tail vein injection twice a week for a total of five times. After administration, the tumor volume of the mice was measured. The parameters were observed and measured regularly. The specific results are shown in Table 5 and Figure 46.

[0401] Conclusion: In the experimental example, the human breast cancer cell line HCC1806 was used to study subcutaneous xenografts of human breast cancer. Dell constructs BT00102 in a mouse tumor-bearing model of HCC1806 human breast cancer. The effectiveness of option 1 was evaluated.

[0402] The experimental results showed that BT001021 (10 mg / kg, IV, BIW × 3W) was HCC18 06 We demonstrated that it can significantly inhibit tumor growth in a xenograft model mouse for breast cancer, thus demonstrating antitumor properties. The activity was superior to that of the positive control Immu-132, which exhibited better antitumor activity.

[0403] [Table 29]

[0404] According to Tables 4 and 5 and Figures 44-46, the antibody drug BT001021 of the present invention is NCI- BT001021 significantly inhibited tumor growth in the N87 mouse model. It is significantly superior to Immu-132 in terms of dosage, and does not cause significant weight loss or significant drug toxicity. In the HCC1806 mouse model, the dosage was reduced due to the high malignancy of the tumor. When the dosage was increased to 10 mg / kg, Immu-132 did not show significant inhibitory activity. However, BT001021 was able to significantly inhibit tumor growth. The result is that BT00102 of the present invention This demonstrated that 1 had high efficacy and excellent safety.

[0405] In the subcutaneous xenograft models of Experimental Examples 1 and 2, the antitumor activity of BT001021 was: At the same dosage, it showed significantly superior antitumor activity compared to Immu-132, which is the case with BT00. 1021 has the potential to treat solid tumors and clinically treats more patients than Immu-132. This suggested that it could be predicted to be beneficial to the individuals involved.

[0406] Experimental Example 3. Inhibition of HCC827 by antibody-drug conjugates Using Experimental Example 3, subcutaneous xenograft human tumor cells from HCC827 non-small cell lung cancer were used. BT001021 and BT001035 for the proliferation of tumor-bearing mouse models constructed using [method / tool] The inhibitory effect was evaluated. Specifically, in the experiment, a tumor-bearing mouse model was used to test for human non-small cell lung Constructed using subcutaneous xenografts of the HCC827 cell line. Tumor volume approximately 100 mm². 3 to After that, the mice were randomly grouped and given BT001021 and BT001035 twice a week. The drug was administered intravenously a total of six times. Subsequently, changes in tumor volume and animal body weight were monitored twice a week. The efficacy of BT001021 and BT001035 against tumor-bearing mice was measured multiple times. The ulcer-inhibiting effect was calculated.

[0407] Experimental method: HCC827 cells were cultured in 1640 culture medium containing 10% fetal bovine serum at 37°C and HCC827 cells were cultured in 5% CO2. Exponential growth phase HCC827 cells were collected and incubated in PBS at an appropriate concentration. The solution was resuspended and subcutaneously inoculated into female Balb / c-nu mice to create a xenograft model for lung cancer. A structure was constructed with an average tumor volume of approximately 80 mm². 3 When this happens, mice are raised according to tumor size. Saline solution group, positive drug Immu-132 (10 mg / kg, IV, BIW x 3W) group , BT001021 (10 mg / kg, IV, BIW × 3W) group and BT001035 ( Students were randomly assigned to one of the following groups (10 mg / kg, IV, BIW x 3 weeks), followed by weekly administration of the corresponding drug. The drug was administered via intravein injection in two separate doses, for a total of six doses. After administration, the tumor volume and body weight of the mice were monitored regularly. The parameters were observed and measured. The results are shown in Table 6, Figure 47A, and Figure 47B.

[0408] Conclusion: The experimental results showed that BT001021 and BT001035 were associated with HCC827 non-small cell lung cancer. It significantly inhibits tumor growth in xenograft model mice, and tumor regression may occur at the end of administration. This demonstrated that the antitumor activity was superior to that of the positive control Immu-132 group. During the observation period, no animal deaths or significant weight loss occurred in any of the treatment groups. No significant drug toxicity was observed. During the treatment period, the mice were given all the drugs evaluated. It showed good tolerability.

[0409] [Table 30]

[0410] According to Table 6, Figures 47A and 47B, BT001021 and BT001035 are both Both showed significant inhibitory activity against tumor growth during the evaluation period, but they were administered at the same dose. It is significantly superior to the inhibitory activity of Immu-132. During treatment, in all groups, significant No weight loss or significant drug toxicity was observed. The results were for BT001021 and BT00 Both 1035 compounds demonstrated excellent antitumor activity.

[0411] In the subcutaneous xenograft model, both BT001021 and BT001035 exhibit antitumor activity. The effect was significantly superior to that of Immu-132 in terms of antitumor activity at the same dose, and BT001 Both 021 and BT001035 have the potential to treat solid tumors, and clinically I This suggests that it may be beneficial to a greater number of patients than mmu-132.

[0412] Experimental Example 4 Inhibition of NCI-N87 by antibody-drug conjugates. Using Experimental Example 4, tumor-bearing mice constructed with subcutaneous xenografts of human tumor cells The inhibition of tumor growth by the antibody-drug conjugate BT001036 was evaluated. Specifically In the experiment, a tumor-bearing mouse model was subjected to subcutaneous xenografts of the human gastric cancer cell line NCI-N87. Therefore, it was constructed. The tumor volume was approximately 140 mm². 3 After that, the mice were randomly grouped and BT 001036 was administered intravenously twice a week for a total of six times. Tumor volume and animal body weight were measured. Changes were measured twice a week to assess the effectiveness of antibody-drug conjugates against tumor-bearing mice (tumor inhibition). The harmful effects were evaluated.

[0413] Experimental method: NCI-N87 cells were cultured in 1640 culture medium containing 10% fetal bovine serum at 37°C and NCI-N87 cells were cultured in 5% CO2. Exponential growth phase NCI-N87 cells were collected and properly cultured in PBS. The solution was resuspended at the appropriate concentration and subcutaneously inoculated into female Balb / c-nu mice to create a xenograft model for gastric cancer. A structure was constructed with an average tumor volume of approximately 140 mm². 3 When this happens, mice are used according to tumor size. The physiological saline group, the BT001036 (1.5 mg / kg, IV, BIW × 3W) group and B Students were randomly assigned to the T001036 (3 mg / kg, IV, BIW × 3W) group, and then, The corresponding drug was administered via tail vein injection twice a week for a total of six times. After administration, the tumor volume of the mice was measured. In addition, their weight was regularly observed and measured. The specific results are shown in Table 7, Figure 48A, and Figure 48B. did.

[0414] [Table 31]

[0415] Conclusion: In the experimental example, a subcutaneous xenograft model of human gastric cancer was created using the human gastric cancer cell line NCI-N87. Constructed using lower xenografts, B in a mouse model of NCI-N87 human gastric cancer The effectiveness of T001036 was evaluated.

[0416] The experimental results showed that high and low doses (1.5 mg / kg, 3 mg / kg) of BT001036 were effective. Both of them exhibited excellent antitumor activity in the tumors of NCI-N87 gastric cancer xenograft model mice. It demonstrated that growth was significantly inhibited and tumor regression may occur at the end of administration. During the observation period, No animal deaths or significant weight loss occurred in any of the treatment groups, and no significant drug toxicity was observed. It was not done. During the treatment period, the mice showed good tolerability for all the drugs evaluated. I showed it.

[0417] Experimental Example 5 Inhibition of MDA-MB-231 by antibody-drug conjugates. Using Experimental Example 5, subcutaneous xenograft human tumor cells of MDA-MB-231 breast cancer were used. The inhibitory effect of BT001021 on tumor growth in a constructed mouse model was evaluated. Specifically, in the experiment, a tumor-bearing mouse model was subjected to the skin of the human breast cancer cell line MDA-MB-231. Constructed using a lower xenograft. Tumor volume was approximately 130 mm². 3 After that, the mice were randomly selected. The patients were grouped together and administered BT001021 intravenously twice a week for a total of six times. Subsequently, the tumor The effectiveness of BT001021 in tumor-bearing mice was determined by measuring changes in tumor volume and animal body weight. The effect (tumor inhibitory effect) was calculated.

[0418] Experimental method: NCI-MDA-MB-231 cells in RPMI164 containing 10% fetal bovine serum. MDA-MB-231 cells were cultured in culture medium at 37°C and 5% CO2. The cells were collected, resuspended to the appropriate concentration in PBS, and subcutaneously affixed to female Balb / c-nu mice. We then constructed a xenograft model of breast cancer. When the average tumor volume reached approximately 130 mm³... The mice were divided into two groups according to tumor size: a saline group and a BT001021 (3 mg / kg) group. Participants were randomly assigned to groups, and then administered the corresponding drug via tail vein injection twice a week for a total of six times. The tumor volume and body weight of the mice were observed and measured regularly after administration. See Table 8 and Figure 4 for the results. This is shown in 9A and Figure 49B.

[0419] Conclusion: The results showed that BT001021 was used in a xenograft model mouse of MDA-MB-231 breast cancer. It demonstrated that tumor growth was significantly inhibited and tumor regression may occur at the end of administration. During the observation period, In all treatment groups, no animal deaths or significant animal weight loss occurred, and no significant drug toxicity was observed. No adverse reactions were observed. During the treatment period, the mice showed good tolerance to all drugs evaluated. It showed tolerance.

[0420] [Table 32]

[0421] In a subcutaneous xenograft model, BT001021 exhibited significant antitumor activity. Observation period Throughout the period, no animal deaths or significant animal weight loss occurred in any of the treatment groups, and no significant drug activity was observed. No physical toxicity was observed. During the treatment period, the mice showed no adverse reactions to all drugs evaluated. They showed good tolerance.

[0422] Example 65: In vivo pharmacokinetic studies of antibody-drug conjugates and bioactive molecules. Using Experimental Example 6, we investigated the pharmacokinetics of antibody-drug conjugates and bioactive molecules in vivo. This was evaluated. Specifically, in the experiment, the tumor-bearing mouse model was compared with Balb / c-nu mice. Constructed by subcutaneous xenografting of the human gastric cancer cell line NCI-N87. Tumor volume was 100 ~200mm 3 After that, the mice were randomly grouped into BT001021 and T-030. A single dose was administered intravenously. The concentrations of T-030 in tumor tissue and serum were determined, and the tumor was identified. In mice with ulcers, the antibody conjugate BT001021 and the bioactive molecule T-030 The pharmacokinetics of mvivo were evaluated.

[0423] Test drug Drug name and preparation method: BT001021, Store liquid alicot at -20°C at a concentration of 20 mg / ml before use. The test solution was obtained by diluting it to the desired dose with physiological saline beforehand; Prepare T-030 with dimethyl sulfoxide to 1 mg / ml and add to physiological saline solution. The test solution was obtained by diluting it to the desired dose. Experimental animals and cell lines: Balb / c-nu mouse (Beijing Vital River Labora Tory Animal Technology Co., Ltd., Production License Number: S CXK(Beijing)2016-0011); Gastric cancer cell line NCI-N87(ATCC ). Experimental group and evaluation method: 100-200mm 3 Tumor-bearing mice (4 mice / group) with a tumor volume were randomly assigned to a group. The group size was determined according to the sample size, and the administration route was a single tail vein injection. .

[0424] Experimental Example 6. Drug activity of BT001021 and T-030 in tumor-bearing mice in vivo Test of attitude Experimental method: NCI-N87 cells were cultured in 1640 culture medium containing 10% heat-inactivated fetal bovine serum. NCI-N87 cells were cultured at 37°C and 5% CO2. Exponential growth phase NCI-N87 cells were collected and incubated in PBS. The mixture was resuspended to the appropriate concentration and subcutaneously inoculated into Balb / c-nu mice to induce xenografts of gastric cancer. Dell was constructed. When the average tumor volume reached approximately 100-200 mm³, the mice were subjected to tumor treatment. Depending on size, the group is divided into the physiological saline group, the T-030 (0.23 mg / kg, IV, single dose) group, and Participants were randomly assigned to either the BT001021 (10 mg / kg, IV, single dose) group, followed by... The corresponding drug was administered by tail vein injection. In the T-030 group, serum and tumor tissue were administered 1 hour after administration. Data was collected at intervals of 2 hours, 4 hours, 8 hours, 24 hours, and 72 hours after administration (T-030 was administered). It was not detected in serum and tumor tissue 72 hours later, therefore, serum and tumor tissue were administered (No samples were collected 168 hours after administration). In the BT001021 group, serum and tumor tissue were collected. The substance was absorbed 1 hour, 2 hours, 4 hours, 8 hours, 24 hours, 72 hours, and 168 hours after administration. The samples were collected, and the concentration of T-030 in serum and tumors was tested by LC-MS / MS. The results are shown in Table 9. The dosage of T-030 (0.23 mg / kg) was compared to the molar dose. I switched to BT001021 (10mg / kg).

[0425] [Table 33]

[0426] Conclusion: AUC of the drug in tumors and serum in the BT001021 (10 mg / kg) dose group last These were 850.1h × ng / ml and 174.97h × ng / ml, respectively, but T AUC of tumors and serum drugs in the -030 treatment group last These are 3.85h × ng respectively. The values ​​were / ml and 5.58h × ng / ml. The comparison was made with the T-0 of the BT001021 dose group. The study showed that the exposure level of 30 was significantly increased compared to the exposure level of the T-030 administration group. Therefore, the exposure level of the bioactive molecule T-030 in tumors in the BT001021 treatment group was compared to the exposure level in serum. Although significantly higher than the dose, the exposure levels of active biomolecules in serum and tumors in the T-030 administration group were below the baseline. Essentially the same, this is because antibody-drug conjugate (BT001021) is highly effective in tumor groups. It demonstrated that it possessed woven targeting properties.

[0427] Bioactive molecule T-03 in tumors and serum in the BT001021 (10 mg / kg) administration group 0 C max The values ​​were 7.82 ng / ml and 11.7 ng / ml, respectively, but T- C of the bioactive molecule T-030 in tumors and serum in the 030 administration group max These are 1.2 The levels were 0 ng / ml and 1.81 ng / ml, which are due to the antibody-drug conjugate (BT0). 01021) showed higher concentrations of the bioactive molecule (T-030) in tumor tissue and serum. It was shown that it possessed this characteristic.

[0428] In tumors treated with BT001021 (10 mg / kg), the bioactive molecule T-030 was observed. T 1 / 2 The average time was 93.14 hours, but the bioactive molecule T in tumors in the T-030 administration group -030 T 1 / 2 This is 2.55 hours, and this is due to the antibody-drug conjugate (BT0 This study showed that 01021) had a longer half-life in tumor tissue.

[0429] In conclusion, BT001021 has advantages compared to its corresponding bioactive molecule (T-030). It exhibited significant tumor tissue targeting properties and favorable pharmacokinetic characteristics.

[0430] Experimental Example 7. In vivo study of antibody-drug conjugates BT001021 and Immu-132 Pharmacokinetic studies. In the experiment, a tumor-bearing mouse model was used to transfer human gastric cancer cell lines (NCI) to Balb / c-nu mice. - Constructed using subcutaneous xenografts of N87. Tumor volume 100-200 mm 3 It became Next, mice were randomly grouped and administered single doses of BT001021 and Immu-132 intravenously. It was administered intravenously. The bioactive molecule T-030 corresponds to BT001021 in tumor tissue and serum. The concentrations of SN-38 and Immu-132 were measured, respectively, to determine the in vivo role of Pharmacokinetics of antibody conjugates BT001021 and Immu-132 in tumor mice I evaluated it.

[0431] Test drug Drug name and preparation method: BT001021, Store liquid alicot at -20°C at a concentration of 20 mg / ml before use. The test solution was obtained by diluting it to the desired dose with physiological saline beforehand; Immu-132 was diluted with physiological saline to the desired dose to obtain the test solution. Experimental animals and cell lines: Balb / c-nu mouse (Beijing Vital River Labora Tory Animal Technology Co., Ltd., Production License Number: S CXK(Beijing)2016-0011); Gastric cancer cell line NCI-N87(ATCC ). Experimental group and evaluation method: 100-200mm 3Tumor-bearing mice (4 mice / group) with a tumor volume were randomly selected. The subjects were grouped (the number of groups was determined according to the sample size), and the route of administration was a single tail vein injection. Ta.

[0432] Experimental method: NCI-N87 cells were cultured in 1640 culture medium containing 10% heat-inactivated fetal bovine serum. NCI-N87 cells were cultured at 37°C and 5% CO2. Exponential growth phase NCI-N87 cells were collected and incubated in PBS. The mixture was resuspended to the appropriate concentration and subcutaneously inoculated into Balb / c-nu mice to induce xenografts of gastric cancer. Dell was constructed. When the average tumor volume reached approximately 100-200 mm³, the mice were subjected to tumor treatment. Depending on size, the BT001021 (5 mg / kg, IV, single dose) group and Immu-1 Participants were randomly assigned to one of 32 groups (5 mg / kg, IV, single dose), followed by the administration of the corresponding drug. It was administered by intravenous injection. Serum and tumor tissue were measured at 2 hours, 24 hours, and 48 hours after administration, respectively. After 72 hours, samples were collected and LC-MS / MS was used to determine T-030 or S in serum and tumors. The concentration of N-38 was tested.

[0433] [Table 34]

[0434] Conclusion: AUC of small toxin molecules in tumors and serum in the BT001021 treatment group last Each The values ​​were 427.2h × ng / ml and 115.3h × ng / ml, but Immu-1 AUC of small toxin molecules in tumors and serum in 32 treatment groups last These are 116.8 each. The values ​​were h × ng / ml and 422.7 h × ng / ml. Tumors in the BT001021 treatment group. The small toxin molecules inside are C maxThe level was 6.8 ng / ml, but in the Immu-132 administration group... Small toxin molecules in tumors max The result was 2.8 ng / ml. 1021 has better tumor tissue targeting and better pharmacokinetics compared to Immu-132. It demonstrated improved behavioral characteristics and a better treatment window.

[0435] Although the specific methods for carrying out the present invention have been described in detail, various changes have been made to the details. Modifications and substitutions may be made in accordance with all published teachings, and such changes may be made in accordance with the present invention. Those skilled in the art should understand that the scope of protection is within the scope of the invention. The entire scope of the invention is covered by the attached patent. The claim and any equivalent thereof are given.

Claims

1. The compound shown in formula (I) below, or a pharmaceutically acceptable salt thereof. T-[L １ -(L ２ ) ｍ１ -(L ３ ) ｍ２ -(L ４ ) ｍ３ -E]-G Formula (I) [In the formula, T is a fragment of a bioactive molecule, preferably a fragment of a molecule having antitumor bioactivity; L １ These include amino acids, peptides composed of 2 to 10 amino acids, oligosaccharides, and -(C) H ２ ) ｔ１ -、-(CH ２ CH ２ O) ｔ１ -(CH) ２ ) ｔ２ - 【Chemistry 1】 Selected from; R, R', R １ and R ２ Each of these independently consists of H (hydrogen), D (deuterium), and H chlorogenic acid group, carboxylic acid group, sulfonic acid group, cyano, C １～６ Alkyl, halogenated C １～６ C substituted with alkyl or cyano １～６ Alkyl (e.g., -CH) ２ CN), C １～６ Alkoxy, C ２～１０ Alkenil, C ２～１０ Alkinyl, C ３～６ Cycloalkyl, Each Z is a 6-10 member aryl or 5-12 member heteroaryl. １ These are independently amino acids or It is a peptide composed of 2 to 10 amino acids, １ and t ２ Each of them is independent x is 0, 1, 2, 3, 4, 5 or 6, １ and x ２ Each of them is independently 0, 1, 2, 3, 4, 5 or 6, and each x ３ L is independently 0, 1, 2, 3 or 4. １ is L １ 1 It is connected to a T at position; L ２ These include amino acids, peptides composed of 2 to 10 amino acids, oligosaccharides, and -(C) H ２ ) ｔ１ -、-(CH ２ CH ２ O) ｔ１ -(CH) ２ ) ｔ２ - 【Chemistry 2】 Selected from; R ３ , R ４ , R ５ and R ６ Each of them is H (hydrogen), D (deuterated hydrogen) (Mu), halogen, carboxylic acid group, sulfonic acid group, CN, C １～６ Alkyl, halogenated C １～６ C substituted with alkyl or cyano １～６ Alkyl, C １～６ Alkoxy, C ２ ～１０ Alkenil, C ２～１０ Alkinyl or C ３～６ Selected independently from cycloalkyl groups either be or R ３ / R ４ , R ５ / R ６ or R ３ / R ５ The carbon atoms bonded to it and They form a 3-8 member ring together, t １ and t ２ Each of them is independently 0, 1, 2, 3, 4 , 5 or 6, y １ and y ２ Each of them independently corresponds to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, L ２ is L ２ In first place １ It is connected to; L ３ This is one or more R ７ The following groups may be substituted: amino, 3- to 8-membered cyclo Alkylene, 3-8 member aliphatic heterocyclylene, 6-12 member bridged heterocyclylene, 6- 12-membered spiroheterocyclylene, 6-12 membered condensed heterocyclylene, 6-10 membered allile Selected from 5-12 member heteroarylenes or 3-8 member cycloalkylenes-W-; W is oxygen or NR ８ And R ７ H (hydrogen), D (deuterium), halogen, = O CN, carboxyl, sulfonic acid group, C １～６ Alkyl, halogenated C １～６ Alkyl , C substituted with cyano １～６ Alkyl, C １～６ Alkoxy, C ２～１０ Alkeni Lu or CC ２～１０ Selected independently from Alkinnil, R ８ H (hydrogen), D (hydrogen) Hmm), C １～６ Alkyl, C ３～６ Cycloalkyl, C １～６ Alkoxy or cyanoC １～２ Independently selected from alkyl, L ３ is L ３ In first place ２ It is connected to; L ４ teeth 【Transformation 3】 Selected from, Z ５ Preferably, C ２～６ Alkenil, C ２～６ Alkinyl, amide The base, sulfuryl, sulfinyl, 6-10 membered arylene, or 5-6 membered heteroarylene. selected from; Z ２ is C １～６ alkylene, C ２～１０ alkenylene, C ２～１０ alkyl Niren, C ３～８ Cycloalkylene, 6-10 membered arylene, or 5-14 membered heteroaryl Selected from Ren; R ９ is H (hydrogen) or C １～６ Selected from alkyl groups; Z ３ It exists not C １～６ alkylene, halogenated C １～６ alkylene or alkoxy-substituted The replaced C １～６ Selected from alkylenes; or R ９ and Z ３ It is connected to it It forms a 4- to 8-membered heterocycline together with the nitrogen atom; α is independently 0, 1, 2 , 3, 4, 5 or 6; L ４ is L ４ It is joined to E in second place; E is one or more R １２ The following groups may be substituted: pyrimidylene, quinolylene Selected from n or pyrolo[2,3-d]pyrimidylene; R １２ H (hydrogen), D (di Deuterium, halogen, CN, nitro, C １～６ Alkyl or halogenated C １～６ a Chosen to be independent from Lukil; G is a leaving group for nucleophilic substitution; I understand １ , m ２ and m ３ Each of them independently corresponds to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or It is 10.

2. L １ が、6a、、、、、、、、、、、、、、、、、、、、、、、、、、 sn, a peptide composed of 2 to 5 amino acids, 【Chemistry 4】 Selected from; R, R', R １ and R ２ Each of them independently consists of H (hydrogen) and D (hydrogen). Rium), C １～６ Alkyl, C ２～１０ Alkenil, C ２～１０ Alkinyl or C ３～ ６ It is a cycloalkyl, Z １ But Val, Cit, Phe, Lys, D-Val, Le u, Gly, Ala, Asn, Val-Cit, Cit-Val, Cit-Ala, Va l-Ala, Lys-Val, Val-Lys(Ac), Phe-Lys, Phe-Ly s(Ac), D-Val-Leu-Lys, Gly-Gly-Arg or Ala-Ala -Asn, x １ x is 0, 1, 2 or 3, ３ is 0, 1, 2, 3 or 4, The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

3. L １ が、6a、、、、、、、、、、、、、、、、、、、、、、、、、、 sn, Cit-Val, Val-Ala, Lys-Val, Val-Lys(Ac), P he-Lys, Phe-Lys (Ac), D-Val-Leu-Lys, Gly-Gly -Arg, Ala-Ala-Asn, 【Transformation 5】 Selected from; R, R' and R １ Each of them independently consists of H (hydrogen) and D (deuterium). ), C １～６ Alkyl, C ２～１０ Alkenil, C ２～１０ Alkinyl or C ３～６ Shik It is a chloroalkyl, Z １ But Val, Cit, Phe, Lys, D-Val, Leu, G ly, Ala, Asn, Val-Cit, Cit-Val, Cit-Ala, Val-A la, Lys-Val, Val-Lys (Ac), Phe-Lys, Phe-Lys (A c), D-Val-Leu-Lys, Gly-Gly-Arg or Ala-Ala-As n is x １ and x ３ Each of them is independently 0, 1, 2, or 3, according to claim 1 or 2. The compound or a pharmaceutically acceptable salt thereof.

4. L １ Yes, Lys, Cit, Cit-Val, Val-Al, Lys-Val, 【Transformation 6】 Selected from; R, R' and R １ Each of them independently consists of H (hydrogen) and D (deuterium). ) or C １～４ It is alkyl, Z １ But Cit, Lys, Cit-Val, Cit-A la, Val-Ala or Lys-Val, x １ and x ３ Each of them is independently 0, The compound according to any one of claims 1 to 3, which is 1 or 2, or the pharmaceutically acceptable thereof. Salt.

5. L １ Yes, Lys, Cit, Cit-Val, Val-Al, Lys-Val, 【Transformation 7】 A compound selected from any one of claims 1 to 4 or a pharmaceutically acceptable compound thereof Salt.

6. L １ but 【Transformation 8】 A compound selected from any one of claims 1 to 5 or a pharmaceutically acceptable compound thereof Salt.

7. L ２ が、6a、、、、、、、、、、、、、、、、、、、、、、、、、、 sn, a peptide composed of 2 to 5 amino acids, 【Chemistry 9】 Selected from; R ３ , R ４ , R ５ and R ６ Each of them is H (hydrogen), D (deuterohydrate) (Mu), halogen, carboxylic acid group, sulfonic acid group, CF ３ , CN, CH ２ CN, C １～４ a Lukil, C １～４ Alkoxy, C ２～６ Alkenil, C ２～６ Alkinyl or C ３～６ C Independently selected from chloroalkyl, y １ and y ２ Each of them independently becomes 0, 1, 2, 3, 4 , 5, 6, 7 or 8, L ２ is L ２ In first place １ It is connected to; I understand １ is 0, 1, 2, or 3. A compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof.

8. L ２ が、6a、、、、、、、、、、、、、、、、、、、、、、、、、、 sn, Val-Cit, Cit-Val, Val-Ala, Lys-Val, Val-L ys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys , Gly-Gly-Arg, Ala-Ala-Asn, 【Chemistry 10】 Selected from; R ３ , R ４ , R ５ and R ６ Each of them is H (hydrogen), D (deuterohydrate) (Mu), halogen, carboxylic acid group, sulfonic acid group, CF ３ , CN, CH ２ CN, C １～４ a Lukil, C １～４ Alkoxy, C ２～６ Alkenil, C ２～６ Alkinyl or C ３～６ C Independently selected from chloroalkyl, y １ and y ２ Each of them independently becomes 0, 1, 2, 3, 4 , 5, 6, 7 or 8, L ２ is L ２ In first place １ It is connected to; I understand １ is 0, 1, or 2. A compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof.

9. L ２ but, 【Chemistry 11】 Selected from; R ３ , R ４ , R ５ and R ６ Each of them is H (hydrogen), D (deuterohydrate) Mu) or C １～４ Selected independently from alkyl, y １ and y ２ Each of them is independently 0, 1 , 2, 3, 4, 5, 6, 7 or 8, L ２ is L ２ In first place １ It is connected to; I understand １ is 1, A compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof.

10. L ２ but, 【Chemistry 12】 A compound selected from any one of claims 1 to 9 or a pharmaceutically acceptable compound thereof Salt.

11. L ２ but 【Chemistry 13】 A compound selected from any one of claims 1 to 10 or a pharmaceutically acceptable compound thereof Salt.

12. L ３ However, one or more R ７ The following groups may be substituted: amino, 3- to 8-membered cyclo Alkylene, 3-8 member aliphatic heterocyclylene, 6-12 member bridged heterocyclylene, 6- 12-membered spiroheterocyclylene, 6-12 membered condensed heterocyclylene, 6-10 membered allile Selected from 5-12 member heteroarylenes or 3-8 member cycloalkylenes-W-; W is oxygen or NR ８ And R ７ However, H (hydrogen), D (deuterium), halogen, = O CF ３ , CN, CH ２ CN, carboxyl, sulfonic acid group, C １～４ Alkyl, C １～ ４ Alkoxy, C ２～６ Alkenyl or C ２～６ Selected independently from Alkinnil; preferred k is the aforementioned 3-8 member aliphatic heterocyclylene, 6-12 member cross-linked heterocyclylene, 6-1 One or more nitrogen atoms in a 2-membered spiroheterocyclylene or a 6- to 12-membered condensed heterocyclylene. Having atoms; preferably, the 3-8 member aliphatic heterocyclylene, 6-12 member bridged heterocyclylene Cyclylene, 6-12 member spiroheterocyclylene, or 6-12 member condensed heterocyclylene Having one or more quaternary nitrogen atoms; preferably, the 3-8 membered aliphatic heterocycline Len, 6-12 membered cross-linked heterocyclylene, 6-12 membered spiroheterocyclylene or 6-1 A two-membered condensed heterocyclylene has one or more nitrogen atoms, and at least one nitrogen atom ga is replaced by =O; R ８ However, H (hydrogen), D (deuterium), C １～６ Alki Ru, C ２～６ Alkenil, C ３～６ Alkinyl, C ３～６ Cycloalkyl, C １～６ Al Coxy or cyanoC １～２ Independently selected from alkyl; I understand ２ is 0, 1, 2, or 3. A compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof.

13. L ３ However, one or more R ７ The following groups may be substituted: amino, 3-6 membered aliphatic groups Selected from heterocyclylene or 5-10 membered heteroarylene; R ７ However, H (hydrogen), D (deuterium), halogen, =O, CF ３ , CN, CH ２ CN, carboxyl, s ulfonic acid group, C １～４ Alkyl, C １～４ Alkoxy, C ２～６ Alkenyl or C ２～６ Independently selected from alkynyls; preferably, the 3-6 membered aliphatic heterocyclylene is 1 It has one or more nitrogen atoms; preferably, one of the 3-6 membered aliphatic heterocyclylenes or having multiple quaternary nitrogen atoms; preferably, the 3-6 membered aliphatic heterocycline It has one or more nitrogen atoms, and at least one nitrogen atom is substituted with =O; I understand ２ is 0, 1, or 2. A compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof.

14. L ３ However, one or more R ７ Select from 5-6 member heteroarylenes which may be substituted with Selected; R ７ However, H (hydrogen), D (deuterium), halogen, =O, CF ３ , CN, CH ２ CN, carboxyl, sulfonic acid group, C １～４ Alkyl, C １～４ Alkoxy, C ２～６ Alkenyl or C ２～６ Selected independently from Alkinnil; I understand ２ is 1, A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof.

15. L ３ However, one or more R ７ The following groups may be substituted: amino, N-methylpiper Selected from lysylene, pyrazolylene, or triazolylene; R ７ However, H (hydrogen), D (di Deuterium, halogen, =O, CF ３ , CN, CH ２ CN, carboxyl, sulfone acid group, C １～４ Alkyl, C １～４ Alkoxy, C ２～６ Alkenyl or C ２～６ Alki Selected independently from Nil; I understand ２ is 0 or 1, A compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof.

16. L ３ Selected from triazolylene; I understand ２ is 0 or 1, A compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

17. L ４ but, 【Chemistry 14】 Selected from, Z ４ R is a 6-10 member arylene or a 5-6 member heteroarylene; １ ０ is H (hydrogen) or C １～６ Selected from alkyl groups; Z ２ However, C １～６ Alkylene, C ２ ～１０ Alkenylene, C ２～１０ Alkinylene or C ３～８ Selected from cycloalkylenes re; R ９ is H (hydrogen) or C １～６ Selected from alkyl groups; Z ３ Either it does not exist or C １ ～６ Selected from alkylenes; or R ９ and Z ３ However, the nitrogen atom bonded to it and Together they form a 4- to 8-membered heterocyclene; α is independently 0, 1, 2, 3, 4, 5 It is 6, L ４ is L ４ It is bonded to E in the second position; L ４ is L ４ It was in second place and joined to E. the law of nature; I understand ３ is 0, 1, 2, or 3. A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

18. L ４ but, 【Chemistry 15】 Selected from, Z ４ R is a benzene ring, １０ is H (hydrogen) or C １～６ From alkyl Selected; Z ２ However, C １～６ Alkylene, C ２～１０ Alkenylene, C ２～１０ Alkini Ren or C ３～８ Selected from cycloalkylenes; R ９ is H (hydrogen) or C １～６ Alki Selected from; Z ３ Either it does not exist or C １～６ Selected from alkylene, or R ９ and Z ３ However, together with the nitrogen atom bonded to it, it forms a 4- to 8-membered heterocyclene. And; α is independently 0, 1, 2, 3, 4, 5 or 6, L ４ is L ４ It was combined with E in second place. L ４ is L ４ It is joined to E in second place; I understand ３ is 0, 1, 2, or 3. The compound according to claim 17 or a pharmaceutically acceptable salt thereof.

19. L ４ but, 【Chemistry 16】 Selected from, Z ４ It is a 5-6 member heteroarylene; R １０ is H (hydrogen) or C １～ ６ Selected from alkyl groups; Z ２ However, C １～６ Alkylene, C ２～１０ Alkenylene, C ２ ～１０ Alkinylene or C ３～８ Selected from cycloalkylenes; R ９ is H (hydrogen) or C １～６ Selected from alkyl groups; Z ３ Either it does not exist or C １～６ Selected from alkylene can be; or R ９ and Z ３ However, together with the nitrogen atom bonded to it, 4-8 member heterozygous It forms rylene; α is independently 0, 1, 2, 3, 4, 5 or 6; L ４ is L ４ of It is tied to E in second place; I understand ３ is 0, 1, 2, or 3. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof.

20. L ４ but, 【Chemistry 17】 Selected from; m ３ A compound according to any one of claims 1 to 19, wherein the value is 1. Pharmaceutically acceptable salts.

21. L ４ but, [Chemistry 18] Selected from; m ３ A compound according to any one of claims 1 to 20, wherein is 1 Pharmaceutically acceptable salts.

22. L ４ but 【Chemistry 19】 Selected from; m ３ A compound according to any one of claims 1 to 21, wherein is 1 Pharmaceutically acceptable salts.

23. E is one or more R １２ Selected from pyrimidilenes which may be substituted with; R １ ２ any of claims 1 to 22, wherein is independently selected from H (hydrogen) or D (deuterium) Any compound listed in item one, or a pharmaceutically acceptable salt thereof.

24. G is a halogen, OMs, OTs, OTf, nitro, or one or more R １３ Place The following groups may be substituted: alkylthio, arylthio, heteroarylthio, alkyl Lusulfinyl, arylsulfinyl, heteroarylsulfinyl, alkylsulfonyl Selected from yl, arylsulfonyl, or heteroarylsulfonyl; R １３ However, H ( Hydrogen, D (deuterium), halogen, CN, nitro, C １～６ Alkyl, halogen cation C １～６ Alkyl, C １～６ Alkoxy, 6-10 membered aryl, or 5-12 membered heteroalkyl A compound or drug according to any one of claims 1 to 23, independently selected from the reel. A scientifically acceptable salt.

25. G is F, Cl, Br, I, OMs, OTs, OTf, methylsulfonyl, ethylsulfonyl Selected from honyl, p-toluenesulfonyl, or naphthalenesulfonyl, claims 1 to A compound or a pharmaceutically acceptable salt thereof as described in any one of item 24.

26. G is F, Cl, Br, OMs, OTs, methylsulfonyl, or p-toluenesulfonyl A compound selected from the above, according to any one of claims 1 to 25, or a pharmaceutically acceptable compound thereof. Salt.

27. The claim is as described in any one of claims 1 to 26, wherein G is selected from Cl or methylsulfonyl. A compound of or a pharmaceutically acceptable salt thereof. 【Request Item 28】 【Chemistry 20】 In this, G is preferably methylsulfonyl and E is preferably pyrimidylene. , m ３ The compound according to any one of claims 1 to 27, or the pharmaceutically acceptable compound thereof, wherein the ratio is 1. Salt. 【Request Item 29】 【Chemistry 21】 but 【Chemistry 22】 And, m ４ Preferably, the integer is between 0 and 6, and the methylsulfonyl is in the pyrimidine ring. A substituent on a carbon atom adjacent to a nitrogen atom, according to any one of claims 1 to 28. A compound or a pharmaceutically acceptable salt thereof. 【Request Item 30】 【Chemistry 23】 but 【Chemistry 24】 And, m ５ Preferably, the integer is between 0 and 6, and the methylsulfonyl is in the pyrimidine ring. A substituent on a carbon atom adjacent to a nitrogen atom, according to any one of claims 1 to 29. A compound or a pharmaceutically acceptable salt thereof. 【Request Item 31】 【Chemistry 25】 but 【Chemistry 26】 And, m ６ Preferably, the integer is between 0 and 6, and the methylsulfonyl is in the pyrimidine ring. A substituent on a carbon atom adjacent to a nitrogen atom, according to any one of claims 1 to 30. A compound or a pharmaceutically acceptable salt thereof. 【Request Item 32】 【Chemistry 27】 but 【Chemistry 28】 And, m ７ The integer is between 1 and 5, and methylsulfonyl is present on the nitrogen atom in the pyrimidine ring. A substituent on an adjacent carbon atom, the compound according to any one of claims 1 to 31 or The pharmaceutically acceptable salt. 【Request Item 33】 【Chemistry 29】 but 【Transformation 30】 And, m ８ The integer is between 1 and 5, and methylsulfonyl is present on the nitrogen atom in the pyrimidine ring. A substituent on an adjacent carbon atom, the compound according to any one of claims 1 to 32 or The pharmaceutically acceptable salt. 【Request Item 34】 【Chemistry 31】 but 【Chemistry 32】 And, m ９ R is an integer between 1 and 5. １３ is hydrogen or C １～６ Selected from alkyl groups, Methylsulfonyl is a substituent on a carbon atom adjacent to the nitrogen atom in the pyrimidine ring. A compound according to any one of claims 1 to 33 or a pharmaceutically acceptable salt thereof. 【Request Item 35】 【Chemistry 33】 but 【Transformation 34】 And, m １０ If is an integer from 0 to 6, Z ４ These are selected from 5-6 member heteroarylenes; Methylsulfonyl is a substituent on a carbon atom adjacent to the nitrogen atom in the pyrimidine ring. A compound according to any one of claims 1 to 34 or a pharmaceutically acceptable salt thereof. 【Request Item 36】 【Chemistry 35】 but 【Transformation 36】 Z ４ However, pyridylene, pyrimidiylene, pyrazolylene, thiazolyylene, oxazoli Selected from lene or triazolylene; methylsulfonyl is the nitrogen atom in the pyrimidine ring. A substituent on a carbon atom adjacent to the compound according to any one of claims 1 to 35 or This is the pharmaceutically acceptable salt. 【Request Item 37】 【Chemistry 37】 but 【Chemistry 38】 Z ４ is selected from oxazolylene or thiazoylene, and methylsulfonyl is, A substituent on a carbon atom adjacent to a nitrogen atom in the limidine ring, any one of claims 1 to 36. The compounds described in item 1 or their pharmaceutically acceptable salts. [Request Item 38] [Chemistry 39] but 【Chemistry 40】 The compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt thereof.

39. - [L １ - (L ２ ) ｍ１ - (L ３ ) ｍ２ - (L ４ ) ｍ３ -E]-G is the next fragment: 【Chemistry 41-1】 【Chemistry 41-2】 【Chemistry 41-3】 【Chemistry 41-4】 A compound according to claim 1 or a pharmaceutically acceptable salt thereof, selected from the above.

40. T is a fragment of a bioactive molecule, and the bioactive molecule is a platinum metal complex (e.g., oxali Metal complexes such as platinum or gold metal complexes; bleomycin or pinyanmycin, etc. Glycopeptide antibiotics; topoisomerase I inhibitors (e.g., camptothecin, hydro Xycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, topotheca (e.g., verotensian or rubitecan) or topoisomerase II inhibitors (e.g., activator) Nomycin D, Adriamycin, Doxorubicin, Duocalmycin, Daunorubicin DNA topoisomers such as mitoxantrone, podophyllotoxin, or etoposide. Fluorose inhibitors; methotrexate, 5-fluorouracil, cytarabine, gemcitabine, Lucaptopurine, pentostatin, fludarabine, cladribine, or narabine, etc. A. Drugs that interfere with synthesis: tubulin inhibitors, vinblastine alkaloids, vincris Structural compounds such as tin, vinblastine, paclitaxel, docetaxel, or cabazitaxel Drugs that act on protein; serine / threonine kinase inhibitors, tyrosine kinase inhibitors tumor cell signaling, such as aspart kinase inhibitors or histidine kinase inhibitors. Pathway inhibitors; proteasome inhibitors; histone desayalase inhibitors; tumor angiogenesis inhibitors Drugs; cyclin inhibitors; meitansine derivatives; calicheamycin derivatives; auristatin Derivatives; pyrrolobenzodiazepine dimer (PBD) derivatives; melphalan; maitomycete C; chlorambucil; or anything that inhibits the proliferation of tumor cells, or inhibits apoptosis or destruction of tumor cells. A compound according to any one of claims 1 to 39, selected from other active substances that promote death A substance or its pharmaceutically acceptable salt.

41. The aforementioned biologically active molecule 【Chemistry 42】 [In the formula, R １４ R １５ Selected from acyl or sulfonyl substituted with R １５ C １～６ Alkyl, halogenated C １～６ Alkyl, 6-10 membered aryl, or 5-12 membered aryl Selected from member heteroaryls; R １６ These are H (hydrogen), D (deuterium), and C １～ ６ Alkyl, or R １７ C is replaced by １～６ Selected from alkyl groups, R １７ This These are not limited to, but include selected aryl or heteroaryls containing phenyl and pyridyl. Selected, m １１ [This value is 0, 1, or 2.] A compound selected from any one of claims 1 to 40 or a pharmaceutically acceptable compound thereof Salt.

42. The aforementioned biologically active molecule 【Chemistry 43】 [In the formula, R １４ R １５ Selected from acyl or sulfonyl substituted with R １５ C １～６ Alkyl, halogenated C １～６ Alkyl, 6-10 membered aryl, or 5-12 membered aryl Selected from member heteroaryls; R １６ These are H (hydrogen), D (deuterium), and C １～ ６ Alkyl, R １７ C is replaced by １～６ Selected from alkyl groups, R １７ is Ari or selected from heteroaryls, m １１ [This value is 0, 1, or 2.] A compound selected from any one of claims 1 to 41 or a pharmaceutically acceptable compound thereof Salt.

43. The aforementioned biologically active molecule 【Chemistry 44】 A compound selected from any one of claims 1 to 42 or a pharmaceutically acceptable compound thereof Salt.

44. The aforementioned biologically active molecule 【Chemistry 45】 A compound selected from any one of claims 1 to 43 or a pharmaceutically acceptable compound thereof Salt.

45. The aforementioned biologically active molecule 【Chemistry 46】 A compound selected from any one of claims 1 to 44 or a pharmaceutically acceptable compound thereof Salt.

46. The aforementioned biologically active molecule 【Chemistry 47】 A compound selected from any one of claims 1 to 45 or a pharmaceutically acceptable compound thereof Salt.

47. The aforementioned biologically active molecule 【Chemistry 48】 A compound selected from any one of claims 1 to 46 or a pharmaceutically acceptable compound thereof Salt.

48. T, 【Chemistry 49-1】 【Chemistry 49-2】 【Chemistry 49-3】 A compound selected from any one of claims 1 to 47 or a pharmaceutically acceptable compound thereof Salt.

49. T, 【Chemistry 50-1】 【Chemistry 50-2】 A compound selected from any one of claims 1 to 48 or a pharmaceutically acceptable compound thereof Salt.

50. T, 【Chemistry 51】 A compound selected from any one of claims 1 to 49 or a pharmaceutically acceptable compound thereof Salt.

51. T, 【Chemistry 52】 A compound selected from any one of claims 1 to 50 or a pharmaceutically acceptable compound thereof Salt.

52. T 【Chemistry 53】 A compound selected from any one of claims 1 to 51 or a pharmaceutically acceptable compound thereof Salt.

53. The aforementioned compound, 【Chemistry 54-1】 【Chemistry 54-2】 【Chemistry 54-3】 【Chemistry 54-4】 【Chemistry 54-5】 【Chemistry 54-6】 【Chemistry 54-7】 【Chemistry 54-8】 【Chemistry 54-9】 【Chemistry 54-10】 A compound selected from any one of claims 1 to 52 or a pharmaceutically acceptable compound thereof Salt.

54. The aforementioned compound, 【Chemistry 55-1】 【Chemistry 55-2】 【Chemistry 55-3】 【Chemistry 55-4】 【Transformation 55-5】 【Chemistry 55-6】 【Chemistry 55-7】 【Transformation 55-8】 A compound selected from any one of claims 1 to 53 or a pharmaceutically acceptable compound thereof Salt.

55. It comprises a bioactive molecule, a linker and a targeting moiety, wherein the targeting moiety is It is linked to the linker via an active group (e.g., a thiol group) to form a conjugate. It is a conjugate.

56. The conjugate according to claim 55, having the structure shown in formula (II) below. {T-[L １ -(L ２ ) ｍ１ -(L ３ ) ｍ２ -(L ４ ) ｍ３ -E]} γ -A Formula (I I) [In the formula, A is the targeting portion (e.g., small molecule ligand, protein, polypeptide or non It is a protein reagent (e.g., sugar, RNA, or DNA); γ is an integer or small integer between 1 and 10. It is a number; preferably, γ is an integer or decimal between 5 and 8 (for example, 5, 6, 7, or 8). ； The remaining bases are as described in any one of claims 1 to 54.

57. The targets of A are epidermal growth factor, Trop-2, CD37, HER2, CD70, and EGFR. vIII, Mesothelin, Folate eceoptor 1, Mucin 1 , CD138, CD20, CD19, CD30, SLTRK6, Nectin 4, Ti ssue factor, Mucin16, Endothelin receptor, STEAP1, SLC39A6, Guanylylcyclase C, PSMA, CC D79b, CD22, Sodium phosphate cotransporter 2B, GPNMB, Trophoblast glycoprotein, AGS-1 6, EGFR, CD33, CD66e, CD74, CD56, PD-L1, TACSTD 2, DR5, E16, STEAP1, 0772P, MPF, Napi3b, Sema 5 b, PSCA hlg, ETBR, MSG783, STEAP2, TrpM4, CRIP TO, CD21, CD79b, FcRH2, NCA, MDP, IL20Rα, Brevi can, EphB2R, ASLG659, PSCA, GEDA, BAFF-R, CD22 , CD79a, CXCR5, HLA-DOB, P2X5, CD72, LY64, FcRH 1, IRTA2, TENB2, integrin α5β6, α4β7, FGF2, FG FR2, Her3, CD70, CA6, DLL3, DLL4, P-cadherin, E pCAM, pCAD, CD223, LYPD3, LY6E, EFNA4, ROR1, SL ITRK6, 5T4, ENPP3, SLC39A6, Claudin18.2, BMPR 1B, E16, STEAP1, Tyro7, 0772P, MPF, Napi3b, Sem a 5b, PSCA hlg, ETBR, MSG783, STEAP2, TrpM4, C RIPTO, CD21, CD79b, FcRH2, NCA, MDP, IL20Rα, Br evican, EphB2R, ASLG659, PSCA, GEDA, CD22, CD7 9a, CXCR5, HLA-DOB, P2X5, CD72, LY64, FcRH1, IR TA2, c-Met, ApoE, CD1 lc, CD40, CD45 (PTPRC), C D49D (ITGA4), CD80, CSF1R, CTSD, GZMB, Ly86, MS 4A7, PIK3AP1, PIK3CD, CCR5, IFNG, IL10RA1, IL- 6, ACTA2, COL7A1, LOX, LRRC15, MCPT8, MMP10, NO G,SERPINEL,STAT1,TGFBR1,CTSS,PGF,VEGFA,C 1QA, C1QB, ANGPTL4, EGLN, ANGPTL4, EGLN3, BNIP 3, AIF1, CCL5, CXCL10, CXCL11, IFI6, PLOD2, KIS S1R, STC2, DDIT4, PFKFB3, PGK1, PDK1, AKR1C1, A KR1C2, CADM1, CDH11, COL6A3, CTGF, HMOX1, KRT3 3A, LUM, WNT5A, IGFBP3, MMP14, CDCP1, PDGFRA, T CF4, TGF, TGFB1, TGFB2, CD1 lb, ADGRE1, EMR2, T NFRSF21, UPK1B, TNFSF9, MMP16, MFI2, IGF-1R, R Claim 55 or 56, selected from NF43, NaPi2b, BCMA, or TENB2 The conjugate described above.

58. A is a folic acid derivative, a glutamate urea derivative, a somatostatin derivative, and an aryl sulfonate. Inamide derivatives (e.g., carbonic anhydrase IX inhibitors) link two aliphatic indoles. Polyenes, cyanine dyes, or small molecule ligands such as IR-783 or its derivatives a conjugate according to any one of claims 55 to 57.

59. A, 【Transformation 56】 A conjugate according to any one of claims 55 to 58, selected from the above.

60. A is an antibody such as a monoclonal antibody or an antigen-binding fragment thereof, and the monoclonal The antibody or its antigen-binding fragment is Fab, Fab', F(ab') ２ , Fd, Fv, dA b. Complementarity-determining fragments, single-chain antibodies (e.g., scFv), non-human antibodies, humanized antibodies, chimeric antibodies Claim comprising an antibody, a fully humanized antibody, a probody, a bispecific antibody, or a multispecific antibody. A conjugate as described in any one of items 55 to 59.

61. A is an anti-Her2 monoclonal antibody or an anti-Trop-2 monoclonal antibody, More specifically, the anti-Trop-2 monoclonal antibody is sacituzumab, M1, M2, or M Selected from 3 antibodies; preferably, the anti-Her2 monoclonal antibody is trastuzma Selected from either bu or pertuzumab; The heavy chain of sacituzumab has the amino acid sequence described in SEQ ID NO: 19; the light chain has the amino acid sequence described in SEQ ID NO: Having the amino acid sequence described in 20; The heavy chain variable region of antibody M1 has the amino acid sequence described in SEQ ID NO: 11; the light chain variable region is Having the amino acid sequence described in SEQ ID NO: 12; The heavy chain variable region of antibody M2 has the amino acid sequence described in SEQ ID NO: 13; the light chain variable region is Having the amino acid sequence described in SEQ ID NO: 14; The heavy chain variable region of antibody M3 has the amino acid sequence described in SEQ ID NO: 15; the light chain variable region is Having the amino acid sequence described in SEQ ID NO: 16; The heavy chain constant regions of antibodies M1, M2, and M3 have the amino acid sequence described in SEQ ID NO: 10; The light chain constant region has the amino acid sequence described in SEQ ID NO:

9. The conjugate according to any one of claims 55 to 60.

62. A is an anti-Her2 monoclonal antibody or an anti-Trop-2 monoclonal antibody, In other words, the anti-Trop-2 monoclonal antibody is selected from sacituzumab, and the anti The Her2 monoclonal antibody is selected from trastuzumab or pertuzumab, claim. A conjugate as described in any one of items 55 to 61.

63. A is an RGD peptide that recognizes cell surface integrin receptors; EGF, PDGF, or or growth factors that recognize cell surface growth factor receptors such as VEGF; or functional cell surface growth factors Rasminogen activator, bombesin, bradykinin, somatostatin, or prostate Selected from peptides capable of recognizing specific membrane antigen receptors, any one of claims 55 to 62 The conjugate described in item 1.

64. A contains CD40 ligand, CD30 ligand, OX40 ligand, PD-1 ligand, ErbB ligand, Her2 ligand, TACSTD2 ligand, or DR5 ligand A conjugate according to any one of claims 55 to 63, selected from among them. 【Request Item 65】 【Chemistry 57-1】 【Chemistry 57-2】 【Chemistry 57-3】 【Chemistry 57-4】 【Chemistry 57-5】 【Chemistry 57-6】 【Chemistry 57-7】 【Chemistry 57-8】 【Chemistry 57-9】 [In the formula, γ is an integer or decimal between 1 and 10, and mAb is anti-Trop-2 monoclonal anti It is either a hermetic antibody or an anti-Her2 monoclonal antibody; preferably, the anti-Trop-2 monoclonal antibody. The monoclonal antibody is selected from sacituzumab, M1, M2, or M3, and the anti-Her2 monoclonal antibody is selected from the anti-Her2 monoclonal antibody. The ronal antibody is selected from trastuzumab or pertuzumab; preferably, γ is 5 to 8 It is an integer or decimal number (for example, 5, 6, 7, or 8). A conjugate according to any one of claims 55 to 64, selected from the above. 【Request Item 66】 【Chemistry 58-1】 【Chemistry 58-2】 【Chemistry 58-3】 【Chemistry 58-4】 【Chemistry 58-5】 【Chemistry 58-6】 【Chemistry 58-7】 [In the formula, γ is an integer or decimal between 1 and 10, and mAb is anti-Trop-2 monoclonal anti It is either a hermetic antibody or an anti-Her2 monoclonal antibody; preferably, the anti-Trop-2 monoclonal antibody. The monoclonal antibody is selected from sacituzumab, and the anti-Her2 monoclonal antibody is trast Selected from zumab or pertuzumab; preferably γ is 5 to 8 (e.g., 5, 6, 7 or 8) is an integer or decimal number. A conjugate according to any one of claims 55 to 65, selected from the above. 【Request Item 67】 【Chemistry 59-1】 【Chemistry 59-2】 【Chemistry 59-3】 【Chemistry 59-4】 [In the formula, A1 is sacituzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal number between 5 and 8. A conjugate according to any one of claims 55 to 66, selected from the above. 【Request Item 68】 【Chemistry 60-1】 【Chemistry 60-2】 [In the formula, A1 is sacituzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 67, selected from the above. 【Request Item 69】 【Chemistry 61】 [In the formula, A1 is sacituzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 68, selected from the above. 【Request Item 70】 【Chemistry 62-1】 【Chemistry 62-2】 【Chemistry 62-3】 【Chemistry 62-4】 [In the formula, A2 is trastuzumab, and γ is an integer or decimal number from 1 to 10; preferably] γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6. It is an integer or decimal between 5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 69, selected from the above. 【Request Item 71】 【Chemistry 63-1】 【Chemistry 63-2】 [In the formula, A2 is trastuzumab, and γ is an integer or decimal number from 1 to 10; preferably] γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6. It is an integer or decimal between 5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 70, selected from the above. 【Request Item 72】 【Chemistry 64】 [In the formula, A2 is trastuzumab, and γ is an integer or decimal number from 1 to 10; preferably] γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6. It is an integer or decimal between 5 and 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 71, selected from the above. 【Request Item 73】 【Chemistry 65-1】 【Chemistry 65-2】 【Chemistry 65-3】 【Chemistry 65-4】 [In the formula, A3 is pertuzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 72, selected from the above. 【Request Item 74】 【Chemistry 66-1】 【Chemistry 66-2】 [In the formula, A3 is pertuzumab, and γ is an integer or decimal number from 1 to 10; preferably, γ is an integer or decimal between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 It is an integer or decimal between 7.5, 6.5 and 8, 7 and 8, or 7.5 and 8. A conjugate according to any one of claims 55 to 73, selected from the above. 【Request Item 75】 【Chemistry 67-1】 【Chemistry 67-2】 【Chemistry 67-3】 【Chemistry 67-4】 [In the formula, A4 is antibody M1, and γ is an integer or decimal number from 1 to 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 5, 6.5-8, 7-8, or 7.5-8. A conjugate according to any one of claims 55 to 74, selected from the above. 【Request Item 76】 【Transformation 68】 [In the formula, A4 is antibody M1, and γ is an integer or decimal number from 1 to 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 5, 6.5-8, 7-8, or 7.5-8. A conjugate according to any one of claims 55 to 75, selected from the above. 【Request Item 77】 【Chemistry 69-1】 【Chemistry 69-2】 【Chemistry 69-3】 【Chemistry 69-4】 [In the formula, A5 is antibody M2, and γ is an integer or decimal number from 1 to 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 5, 6.5-8, 7-8, or 7.5-8. A conjugate according to any one of claims 55 to 76, selected from the above. 【Request Item 78】 【Chemistry 70】 [In the formula, A5 is antibody M2, and γ is an integer or decimal number from 1 to 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 5, 6.5-8, 7-8, or 7.5-8. A conjugate according to any one of claims 55 to 77, selected from the above. 【Request Item 79】 【Chemistry 71-1】 【Chemistry 71-2】 【Chemistry 71-3】 【Chemistry 71-4】 [In the formula, A6 is antibody M3, and γ is an integer or decimal number from 1 to 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 5, 6.5-8, 7-8, or 7.5-8. A conjugate according to any one of claims 55 to 78, selected from the above. [Request Item 80] [Chemistry 72] [In the formula, A6 is antibody M3, and γ is an integer or decimal number from 1 to 10; preferably, γ is , integers or decimals between 5 and 8, for example, 6 to 7, 6 to 7.5, 6 to 8, 6.5 to 7, 6.5 to 7 It is an integer or decimal between 5, 6.5-8, 7-8, or 7.5-8. A conjugate according to any one of claims 55 to 79, selected from the above.

81. A method for preparing a conjugate according to any one of claims 55 to 80. The linker of the compound of formula (I) is coupled with the active group of the targeting portion. A method that includes steps.

82. The linker of the compound of formula (I) is coupled with the active group of the targeting portion to C The method according to claim 81, comprising the step of forming an S bond.

83. The targeting portion of the conjugate is an anti-Her2 monoclonal antibody or or an anti-Trop-2 monoclonal antibody or its active fragment or variant; preferred or the anti-Trop-2 monoclonal antibody is sacituzumab, M1, M2, or M3 Selected from antibodies, the anti-Her2 monoclonal antibody is trastuzumab or pertuzumab The method according to claim 81 or 82, selected from B.

84. The targeting portion of the conjugate is an anti-Her2 monoclonal antibody or or an anti-Trop-2 monoclonal antibody or its active fragment or variant; preferred The anti-Trop-2 monoclonal antibody is selected from sacituzumab, and the anti-He Claim 81, the r2 monoclonal antibody is selected from trastuzumab or pertuzumab. The method described in any one of items ~83.

85. The molar ratio of the targeting portion of the conjugate to the compound of formula (I) is 1 : (1 to 20); preferably, the coupling is carried out in water and / or organic solvents. Preferably, the organic solvent is N,N-dimethylformamide, dimethyl sulfopropyl alcohol. Sides, N-methylpyrrolidone, nitriles (e.g., acetonitrile), alcohols (e.g., Claim 81, selected from methanol, ethanol, or any combination thereof The method described in any one of items ~84.

86. The process further comprises a step of purifying the coupling product; preferably, the coupling The product is purified by chromatography; preferably, the chromatography is Ion exchange chromatography, hydrophobic chromatography, reversed-phase chromatography or any of claims 81 to 85, comprising one or more affinity chromatography The method described in any one of the items.

87. A compound according to any one of claims 1 to 54 or a pharmaceutically acceptable salt thereof A conjugate according to any one of claims 55 to 80, and one or more pharmaceuticals A pharmaceutical composition containing additives.

88. In the manufacture of pharmaceuticals for treating diseases associated with abnormal cell activity (e.g., cancer) , the compound described in any one of claims 1 to 54 or a pharmaceutically acceptable salt thereof or The conjugate according to any one of claims 55 to 80 or the medical device according to claim 87 Use of pharmaceutical compositions.

89. Claims 1 to 54 for treating diseases associated with abnormal cell activity (e.g., cancer) Any compound described in one of the claims or a pharmaceutically acceptable salt thereof, or the compound described in any of claims 55 to 80 Use of the conjugate described in any one of the claims or the pharmaceutical composition described in claim 87.

90. The aforementioned cancers include esophageal cancer (e.g., esophageal adenocarcinoma, esophageal squamous cell carcinoma), brain tumors, lung cancer ( For example, small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer Hmm, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer Liver cancer, kidney cancer, non-Hodgkin lymphoma, central nervous system tumors (e.g., glioma, polycystic tumors) Solid tumors or non-solid tumors such as glioblastoma, glioma, or sarcoma, prostate cancer, and thyroid cancer. Use according to claim 88 or 89, selected from tumors.

91. A method for treating abnormal cell activity and associated diseases (e.g., cancer), and a claim for an effective amount. A compound described in any one of items 1 to 54 or a pharmaceutically acceptable salt thereof, or claim The conjugate described in any one of claims 55 to 80 or the pharmaceutical composition described in claim 87 A method comprising the step of administering a substance to an individual that needs it.

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