Conjugates of a cell-binding molecule with camptothecin analogs

Abstract

This invention relates to a conjugate of a camptothecin analogue and a cell-binding molecule of formula (I), wherein R1, R2, R3, R4, R5, X, L, n, m, T and as defined. The invention also relates to methods of using compositions containing the conjugate of a camptothecin analogue and a cell-binding molecule in the treatment of cancer, autoimmune diseases, and infectious diseases.

Description

Technical Field

[0001] This invention relates to conjugates of camptothecin analogues and cell surface receptor-binding molecules for targeted therapy. The invention also relates to the use of compositions containing conjugates of camptothecin analogues and cell-binding molecules in the treatment of cancer, autoimmune diseases, and infectious diseases. Prior Technology

[0002] Targeted cancer therapy aims to minimize or eliminate side effects by better targeting drugs to tumors and avoiding healthy tissue. One strategy is antibody-drug conjugates (ADCs), which use conditionally stable linkers to bind antibodies that precisely recognize tumors to highly potent cytotoxic drugs (the payload), thereby increasing the local concentration of the latter in the tumor (several times higher than in healthy tissue). Over the past four decades, the pharmaceutical industry has invested heavily in and conducted in-depth research on ADCs. The US FDA has approved eight ADC drugs, namely Mylotarg (gemtuzumab ozogamicin), Adcetris (brentuximab vedotin), Kadcyla (ado-trastuzumab emtansine), Besponsa (inotuzumab ozogamicin), Polivy (polatuzumab vedotin-piiq), Enhertu (fam-trastuzumab deruxtecan-nxki), Padcev (enfortumab vedotin-ejfv), and Trodelvy (sacituzumab govitecan). More than 100 new ADC drugs are in clinical development (Chau, CH et al., Lancet. 2019, 394, 793-804).

[0003] It is well known that the payload-linker components in an ADC complex have a significant impact on the homogeneity of the ADC, its stability in the circulatory system, its pharmacokinetic characteristics, tolerability, and overall therapeutic efficacy (Zhao, RY et al. (2011) J. Med. Chem. 54, 3606; Acchionea, M. et al. (2012) mAbs, 4, 362; Doronina, S. et al. (2006) Bioconjug Chem. 17, 114; Hamann, P. et al. (2005) Bioconjug Chem. 16, 346). Despite extensive research efforts to improve these parameters of ADCs, the chosen payloads to date remain largely derived from maytansin (DM1 and DM4), auristatin (MMAE and MMAF), calicheamicin, pyrrolo[2,1-c][1,4]benzodiazepine (PBD) dimer, camptothecin, duocarmysin, and tubulysin (Leung, D., et al., Antibodies (Basel). 2020, 9: E2. doi: 10.3390 / antib9010002; Khongorzul, P., et al., Mol Cancer Res. 2020, 18: 3-19. doi: 10.1158 / 1541-7786.MCR-19-0582; Chau, CH, et al., Lancet. 2019, 394:793-804. doi: 10.1016 / S0140-6736(19)31774-X).

[0004] Among these payloads, ADCs constructed with camptothecin (CPT) have been shown to have a wider therapeutic index (TI). Numerous clinical trials have demonstrated that two of these ADC compounds, Enhertu (fam-trastuzumab deruxtecan-nxki or DS-8201a) and Sacituzumab govitecan (IMMU-132 or hRS7-SN-38), have significant clinical benefits (PFS and OS) in the treatment of solid tumors (Pondé, N., et al., Curr Treat Options Oncol. 2019 Apr 1;20(5):37. doi: 10.1007 / s11864-019-0633-6; Kaplon, H., et al., MAbs. 2020, 12(1): 1703531, 10.1080 / 19420862.2019.1703531). Camptothecin (CPT) is a potent antitumor agent, successfully isolated from an extract of the native Chinese tree *Camptotheca acuminata* in 1958, and has been widely used in traditional Chinese medicine for centuries. Camptothecin can induce cell death by interacting with the DNase topoisomerase I to form a reversible enzyme-camptothecin-DNA ternary complex (Wu Du, Tetrahedron 59 (2003) 8649-8687). Many camptothecin analogues have been disclosed in the past fifty years, as shown below: Camptothecin, 20-Deoxycamptothecin 10-Methoxycamptothecin, 9-Methoxycamptothecin, 10-Hydroxycamptothecin, 19-Dehydrocamptothecin, Topotecan, Irinotecan (camtosar) Simmitecan SN-38 Silatecan Karenitecin Iciten, Lurtotecan GI-149893 9-Glycine-nitroglycerin camptothecin hydrochloride (MLS000756803) 10,11-Methylenedioxycamptothecin (Mdo-CPT) Gemmanotecan Belotecan Rubitecan or IDEC-132 analogues Elomotecan (BN-80927) 9-Aminocamptothecin (9-AC) 10-Aminocamptothecin (10-AC) 11-Cyanocamptothecin [] [(]NSC609951), 7-Acetyl-camptothecin (SCHEMBL6851483) 7-Hydroxymethylcamptothecin, 10-Amino-11-hydroxycamptothecin [(]BDBM50285230) Diflomotecan, Camptothecin 20-O-propionate (CZ-48) Camptothecin chloroacetate (DL5FFZ93W4) Camptothecin Lysine Ester (NSC610457) Sacituzumab govitecan (Immu-132) (mAb is a Trop2-targeting monoclonal antibody) Trastuzumab deruxtecan (DS-8201a) (mAb is a Her2-targeting monoclonal antibody, trastuzumab) (Jeffrey S. et al. US2019 / 0343828), (Li W. et al., ACS Med Chem Lett. 2019, 10(10): 1386–1392).

[0005] Because camptothecin (CPT) and most of its derivatives are extremely poorly soluble in physiological buffer solutions, the rate of adverse drug reactions was very high in the initial clinical trials conducted since the 1970s. The aggregation of camptothecin-based ADCs can be as high as 80% (Burke, P. et al. Bioconjugate Chem. 2009, 20, 6, 1242-1250), thus limiting its large-scale production. The systemic side effects caused by the aggregates also limit the conduct of clinical trials. To date, the US FDA has approved only three water-soluble CPT derivatives for cancer chemotherapy, including topotecan, irinotecan, and berotetane (Palakurthi, S., Expert Opin Drug Deliv. 2015;12(12):1911-21; Shang, XF et al., Med Res Rev. 2018, 38(3):775-828) and one water-soluble CPT derivative-conjugated ADC, Enhertu (fam-trastuzumab deruxtecan-nxki or DS-8201a), for targeted immunotherapy of Her2 solid tumors (Modi S et al., N Engl J Med.2020, 382(7): 610-621. doi: 10.1056 / NEJMoa1914510; Keam, SJ, Drugs. 2020 Apr;80(5):501-508). doi: 10.1007 / s40265-020-01281-4). We have been researching water-soluble CPT derivative ADCs for some time, using hydrophilic branched linkers to couple with CPT analogs (see PCT / CN2019 / 092614) to expand the therapeutic window of the generated ADC. This invention discloses a water-soluble CPT derivative ADC, wherein the C-10 position of the CPT analog is connected to O or NH, which is crucial for improving water solubility, while the C-11 position is connected to an electron-withdrawing group or a larger group, which can maintain high cytotoxicity comparable to that of natural CPT. Summary of the Invention

[0006] This invention includes conjugates of camptothecin analogues and cell-binding molecules, camptothecin analogue-linkers, and camptothecin analogues, methods for their preparation and use, and intermediates used in their preparation. The camptothecin analogues of this invention are water-soluble and stable in the blood circulation. Once the metabolites of the camptothecin analogue or camptothecin analogue-linker on the conjugate are released from or near disordered cell growth, they can induce cell death.

[0007] In one embodiment, the conjugate of camptothecin analogue has a structure as shown in formula (I): (I) Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polymorphic crystal structure thereof; or an isotope, optical isomer, racemate, non-mirror image isomer, or mirror image isomer thereof; Where T is the target or binding ligand; L is the releasable linker; It is a bond that independently connects L to the atoms of R1, R2, R3, or R5 within the parentheses; n is 1-30, m is 1-10; The part in parentheses is an active camptothecin analogue, in which: R1 and R2 are independently H; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium), aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy, alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently H, C(O)NH, C(O)O, SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, C(O)R6, C(O)NHR6; C1-C6 straight-chain or branched alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl acetylamine, oxime; C5-C12 glycoside, NH2 or OH; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NHR6, N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is H, C(O)O, C(O)NH, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide, or amino acid; or pharmaceutically acceptable salts; Furthermore, R1, R2, R3, and R6 can be omitted independently, and together with R2, R3, X, C-10, and C-9, they can form 5-, 6-, or 7-membered heterocycles.

[0008] In another embodiment, the linker L of the camptothecin analogue and the binding molecule has the following structure: --Ww-(Aa)r-Vv--; where: --W-- is an extension unit; w is 0 or 1; each -Aa- is independently an amino acid unit; r is independently an integer from 0 to 12; --V-- is a spacer unit; v is 0, 1, or 2. The extension W independently comprises a self-destructing spacer, a peptide unit, a hydrazone bond, a disulfide bond, or a thioether bond.

[0009] In another embodiment, the cell surface binding molecule T can be any type of cell-binding ligand, currently known or soon to be known, such as peptides and non-peptides. Typically, the binding molecule T is an antibody; a single-chain antibody; an antibody fragment binding to a target cell; a monoclonal antibody; a single-chain monoclonal antibody; a monoclonal antibody fragment binding to a target cell; a chimeric antibody; a chimeric antibody fragment binding to a target cell; a domain antibody; a domain antibody fragment binding to a target cell; adnectin-like antibodies; DARPins; lymphocyte factors; hormones; vitamins; growth factors; colony-stimulating factors; nutrient transport molecules (transferrin); and peptides, proteins, antibodies, or small molecules bound to albumin, polymers, dendritic macromolecules, liposomes, nanoparticles, vesicles, or (viral) capsids. A preferred binding molecule T is a monoclonal antibody.

[0010] In another case, the compound of formula (I) or its pharmaceutically acceptable salt or solvate may be used to treat cancer, autoimmune diseases or infectious diseases in humans or animals. Simple Explanation of the Diagram

[0011] Figure 1 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 2 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 3 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 4 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 5 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 6 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 7 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 8 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 9 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 10 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 11 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 12 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 13 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 14 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 15 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 16 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 17 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 18 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 19 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 20 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 21 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 22 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 23 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 24 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 25 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 26 shows the synthesis of camptothecin analogues containing conjugable linkers and their conjugation with antibodies. Figure 27 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 28 shows the synthesis of the coupling linker used to prepare camptothecin analogue couplings. Figure 29-1 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 29-2 shows the structure of camptothecin analogues used in coupling agents. Figure 30 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 31 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 32 shows the synthesis of camptothecin analogues containing coupling linkers. Figure 33 shows the structure of the antibody-camptothecin analog conjugate. Figure 34 shows a comparison of the in vivo antitumor effects of antibody-CPT derivative conjugates targeting Her2, C1-031, C1-238, C1-397, C1-407, C1-411, C1-414, C1-424 and C1-428, with T-DM1 in a human gastric tumor N87 cell model, administered intravenously at a dose of 6 mg / kg. Figure 35 shows a comparison of antibody-CPT derivative conjugates targeting Her2, C1-031, C1-226, C1-238, C1-397, C1-407, C1-411, C1-414, C1-424, and C1-428, with T-DM1, administered intravenously at a dose of 150 mg / kg. Figure 36 shows the antitumor effects of EGFR-targeting antibody-CPT derivative conjugates C1-031, C1-200, C1-214, C1-226, C1-305, C1-306, C1-311, C1-362, C1-402, C-407 and C1-419 in a human NSCLC tumor HCC827 cell model, administered intravenously at a dose of 6 mg / kg. Implementation

[0012] definition "Alkyl" refers to an aliphatic hydrocarbon group or monovalent group formed by removing one or two hydrogen atoms from an alkane. It can be straight-chain or branched, and has C1-C8 (1-8 carbon atoms) in the chain. "Branched" means that one or more low-carbon alkyl groups, such as methyl, ethyl, or propyl, are attached to a straight-chain alkyl group. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tributyl, n-pentyl, 3-pentyl, octyl, nonyl, decyl, cyclopentyl, cyclohexyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 3,3-dimethylpentyl, 2,3,4-trimethylpentyl, 3-methylhexyl, 2,2-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 3,5-dimethylhexyl, 2,4-dimethylpentyl, 2-methylheptyl, 3-methylheptyl, n-heptyl, isoheptyl, n-octyl, and isooctyl. C1-C8 alkyl groups may be unsubstituted or substituted with one or more groups, including but not limited to C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH 2, -C(O)NHR', -C(O)N(R') 2, -NHC(O)R', -SR', -S(O) 2R', -S(O)R', -OH, -halogen, -N 3, -NH 2, -NH(R'), -N(R') 2 and -CN; wherein each R' is independently selected from the group consisting of C1-C8 alkyl and aryl groups.

[0013] "Halogen" refers to fluorine, chlorine, bromine, or iodine atoms; preferably fluorine and chlorine atoms.

[0014] "Heteroalkyl" refers to a C2-C8 alkyl group in which one to four carbon atoms are independently replaced by heteroatoms selected from the group of O, S and N.

[0015] "Carbon ring" refers to a saturated or unsaturated monocyclic ring containing 3 to 8 carbon atoms, or a saturated or unsaturated bicyclic ring containing 7 to 13 carbon atoms. Monocyclic carbon rings have 3 to 6 ring atoms, typically 5 or 6. Bicyclic carbon rings have 7 to 12 ring atoms, forming bicyclic systems of [4,5], [5,5], [5,6], or [6,6], or have 9 or 10 ring atoms, forming bicyclic systems of [5,6] or [6,6]. Representative C3-C8 carbon rings include, but are not limited to: -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptanetrienyl, -cyclooctyl, and -cyclooctadienyl.

[0016] "C3-C8 carbon ring" refers to a 3-, 4-, 5-, 6-, 7-, or 8-membered saturated or unsaturated non-aromatic carbon ring. The C3-C8 carbon ring may be unsubstituted or substituted with one or more groups, including but not limited to C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2, -NHC(O)R', -SR', -S(O)R', -S(O)2R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN; wherein each R' is independently selected from the group consisting of C1-C8 alkyl and aryl groups.

[0017] "Alkenyl" refers to a straight-chain or branched aliphatic hydrocarbon group containing carbon-carbon double bonds, with 2-8 carbon atoms in the chain. Exemplary alkenyl groups include vinyl, propenyl, n-butenyl, isobutenyl, 3-methylbut-2-enyl, n-pentenyl, hexenyl, heptenyl, and octenyl.

[0018] "Alynyl" refers to an aliphatic hydrocarbon group containing a carbon-carbon triple bond, which can be a straight chain or a branched chain having 2 to 8 carbon atoms. Exemplary alkynyl groups include ethynyl, propynyl, n-butynyl, 2-butynyl, 3-methylbutynyl, 5-pentynyl, n-pentynyl, hexynyl, heptenyl, and octyynyl.

[0019] "Alkyl group" refers to a saturated branched, straight, or cyclic hydrocarbon group containing 1-18 carbon atoms, and carrying two monovalent free radicals formed by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkane. Typical alkyl groups include, but are not limited to: methylene (-CH 2-), 1,2-ethyl (-CH 2CH 2-), 1,3-propyl (-CH 2CH 2CH 2-), 1,4-butyl (-CH 2CH 2CH 2CH 2-), etc.

[0020] "Alkenyl" refers to an unsaturated branched, straight, or cyclic hydrocarbon group containing 2-18 carbon atoms, and carrying two monovalent free radicals generated by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkene. Typical alkenyl groups include, but are not limited to: 1,2-ethylene (-CH=CH-).

[0021] "Imyynyl" refers to an unsaturated branched, straight, or cyclic hydrocarbon group containing 2-18 carbon atoms, and carrying two monovalent free radicals formed by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkynyl group. Typical ynylyl groups include, but are not limited to: acetylene, propynyl, and 4-pentynyl.

[0022] "Aryl" or "aromatic group" refers to an aromatic or heteroaromatic group consisting of one or more rings, containing three to fourteen carbon atoms, preferably six to ten carbon atoms. The term "heteroaromatic group" refers to a group formed by replacing one or more carbon atoms, preferably one, two, three or four carbon atoms, of an aromatic group with oxygen (O), nitrogen (N), silicon (Si), selenium (Se), phosphorus (P) or (S), preferably with oxygen, sulfur and nitrogen. The term "aryl" or "aromatic group" also refers to an aromatic group formed by the independent substitution of one or more hydrogen atoms by -R', halogen, -OR', -SR', -NR'R'', -N=NR', -N=R', -NR'R'', -NO2, -S(O)R', -S(O)2R', -S(O)2OR', -OS(O)2OR', -PR'R'', -P(O)R'R'', -P(OR')(OR''), -P(O)(OR')(OR''), or -OP(O)(OR')(OR''). R' and R'' are independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, aralkyl, carbonyl, or their pharmaceutically acceptable salts.

[0023] "Heterocyclic" refers to a ring structure in which one to four carbon atoms are independently replaced by heteroatoms such as O, N, S, Se, B, Si, or P. O, N, and S are preferred heteroatoms. A description of heterocyclic compounds can also be found on pages 225-226 of the 78th edition of the *Handbook of Chemistry and Physics* (CRC Press, Inc., 1997-1998), which is cited here for reference. Preferred non-aryl heterocyclic compounds include epoxy, aziridinyl, thiopropyl, pyrrolidinyl, pyrazolidinyl, imidazoridinyl, ethylene oxide, tetrahydrofuranyl, dioxalyl, tetrahydropyranyl, dioxalyl, dioxalyl, piracetinyl, piracetinyl, morpholinyl, piracetinyl, imidazolinyl, pyrrolidinyl, pyrazolidinyl, thiazolinyl, tetrahydrothiaranyl, dithiaranyl, thiomorpholinyl, dihydropiracetinyl, tetrahydropiracetinyl, dihydropiracetinyl, tetrahydropyridinyl, dihydropyridinyl, tetrahydropyrimidinyl, dihydrothiaranyl, azirheptanyl, and fused ring systems obtained by condensing the above groups with phenyl groups.

[0024] The term "heteroaryl" or "aryl heterocycle" refers to an aromatic heterocycle containing 3 to 14, preferably 5 to 10 atoms, comprising monocyclic, bicyclic, or polycyclic compounds. Examples include pyrroleyl, pyridyl, pyrazolyl, thiopheneyl, pyrimidinyl, pyrazinyl, tetrazolyl, indolyl, quinolinyl, purinyl, imidazolyl, thiopheneyl, thiazolyl, benzothiazolyl, furanyl, benzofuranyl, 1,2,4-thiadiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoquinolinyl, benzothiapheneyl, isobenzofuranyl, pyrazolyl, zozolyl, benzimidazolyl, isoxazolyl, pyridyl-N-oxide, and fused-ring systems obtained by condensing the above groups with phenyl groups.

[0025] "alkyl", "cycloalkyl", "alkenyl", "alkynyl", "aryl", "heteroaryl", "heterocyclic", etc., also include their respective "extrinyl", "cycloalkylene", "alkenyl", "alkynylene", "aryl", "heteroaryl", "heterocyclic", etc. For the sake of convenience, this patent application does not make a complete distinction between them.

[0026] "Aryl group" refers to a class of noncycloalkyl groups in which one of the hydrogen atoms bonded to a carbon atom (usually the terminal or sp3 carbon atom) is replaced by an aryl group. Typical aryl groups include benzyl, 2-phenylethyl-1-yl, 2-phenylvinyl-1-yl, naphthylmethyl, 2-naphthylethyl-1-yl, 2-naphthylethyl-1-yl, naphthobenzyl, 2-naphthylphenyl-1-yl, etc.

[0027] "Heteroarylene" refers to a class of noncyclic alkyl groups in which one of the hydrogen atoms bonded to a carbon atom (usually the terminal or sp3 carbon atom) is replaced by a heteroaryl group. Examples of heteroarylene groups include 2-benzimidazolylmethyl and 2-furanylethyl.

[0028] Examples of “hydroxyl protecting groups” include methoxymethyl ethers, 2-methoxyethoxymethyl ethers, tetrahydroguanylate ethers, benzyl ethers, p-methoxybenzyl ethers, trimethylsilyl ethers, triethylsilyl ethers, triisopropylsilyl ethers, tributyldimethylsilyl ethers, triphenylmethylsilyl ethers, acetates, substituted acetates, pivalate esters, benzoate esters, methanesulfonate esters, and p-toluenesulfonate esters.

[0029] A “leaving group” refers to a functional group that can be replaced by another functional group. These leaving groups are well-known in the industry, and examples include halides (such as chlorides, bromides, and iodides), methanesulfonyl, p-toluenesulfonyl, and trifluoromethanesulfonyl. Preferred leaving groups are selected from the group consisting of: nitrophenoxy; N-hydroxysuccinimino (NHS); phenoxy; dinitrophenoxy; pentafluorophenoxy; tetrafluorophenoxy; difluorophenoxy; monofluorophenoxy; pentachlorophenoxy; trifluoromethanesulfonyl; imidazolyl; chlorophenoxy; tetrachlorophenoxy; 1-hydroxybenzotriazolyl; toluenesulfonyl; methanesulfonyl; 2-ethyl-5-phenylisoxazole-3'-sulfonyl; anhydrides or anhydrides formed by the reaction of anhydrides with other anhydrides, such as acetic anhydride or formic anhydride; or intermediates formed by the reaction of polypeptide condensation reagents or Mitsunobu reagents.

[0030] The following abbreviations are used in this invention and are defined as follows: Boc, tributoxycarbonyl; BroP, bromotetradecylphosphonium hexafluorophosphate; CDI, 1,1'-carbonyldiimidazole; DCC, dicyclohexylcarbodiimide; DCE, dichloroethane; dichloromethane; DEAD, diethyl azodicarbonate; DIAD, diisopropyl azodicarbonate; DIBAL-H, diisobutylaluminum hydride; DIPEA or DEA, diisopropylethylamine; DEPC, diethylcyanophosphate; DMA, N,N-dimethylacetamide; DMAP, 4-(N,N-dimethylamino)pyridine; DMF, N,N-dimethylmethamide; DMSO, dimethyl sulfoxide; DTPA, diethylenetriaminepentaacetic acid; DTT, dithiothreitol; EDC, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; ESI-MS, electrospray ionization mass spectrometry; ethyl acetate; Fmoc, N-(9-pyromethoxycarbonyl); HATU, O- (7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate; HOBt, 1-hydroxybenzotriazole; HPLC, high performance liquid chromatography; NHS, N-hydroxysuccinimide; MeCN, acetonitrile; methanol; MMP, 4-methylmorpholine; PAB, p-aminobenzoic acid; PBS, phosphate buffer (pH 7.0-7.5); Ph, phenyl; Phe, L-phenylalanine; PyBrop, bromo-tri-pyrrolidine-phosphonium hexafluorophosphate; PEG, polyethylene glycol; SEC, size exclusion chromatography; TCEP, tris(2-carboxyethyl)phosphine; trifluoroacetic acid; THF, tetrahydrofuran; Val, valine; TLC, thin layer chromatography; UV, ultraviolet light.

[0031] "Amino acids" can be natural or non-natural, with α-amino acids being preferred. Natural amino acids can be encoded by the genetic code; they are alanine, arginine, aspartic acid, cysteine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine, tryptophan, and valine. Non-natural amino acids are derivatives of protein amino acids, including hydroxyproline, lanethionine, 2-aminoisobutyric acid, dehydroalanine, γ-aminobutyric acid (a neurotransmitter), ornithine, citrulline, β-alanine (3-aminopropionic acid), γ-carboxyglutamic acid, selenocysteine ​​(found in many non-eukaryotic and most eukaryotic cells, but not directly encoded by DNA), pyrrolidone (found only in some archaea and one bacterium), N-methoxymethionine (often the original amino acid in proteins in bacteria, mitochondria, and chloroplasts), 5-hydroxytryptophan, L-dihydroxyphenylalanine, triiodothyronine, L-3,4-dihydroxyphenylalanine (DOPA), and O-phosphoserine. The term "amino acid" also includes amino acid analogs and simulants. Analogs are compounds having the same general structural formula as natural amino acids, H₂N(R)CHCO₂H, where R is in the natural amino acid. Examples of analogues include homoserine, ortholeucine, 3-aminopropionic acid, 4-aminobutyric acid, 5-aminovaleric acid, 6-aminohexanoic acid, 7-aminoheptanoic acid, methionine-sulfuric acid, and methionine methyl strontium. More preferred are amino acid mimics, which are compounds with different chemical structures from α-amino acids but similar modes of action. Natural amino acids are mostly in the "L" stereochemical configuration, while "non-natural amino acids" are also used to represent amino acids in the "D" configuration. When using 1 to 8 amino acids in this patent application, their sequences are preferably sequences recognizable by proteases. Many hydrolases have known recognition sequences in the field; see: Matayoshi et al. Science 247: 954 (1990); Dunn et al. Meth. Enzymol. 241: 254 (1994); Seidah et al. Meth. Enzymol. 244: 175 (1994); Thornberry, Meth. Enzymol. 244: 615 (1994); Weber et al. Meth. Enzymol. 244: 595 (1994); Smith et al. Meth. Enzymol. 244: 412 (1994); and Bouvier et al. Meth. Enzymol. 248: 614 (1995); cited herefor reference.Specifically, the following sequences are selected from the following groups: Val-Cit, Ala-Val, Ala-Ala, Val-Val, Val-Ala-Val, Lys-Lys, Ala-Asn-Val, Val-Leu-Lys, Cit-Cit, Val-Lys, Ala-Ala-Asn, Asp-Lys, Asp-Glu, Glu-Lys, Lys, Cit, Ser, and Glu.

[0032] A glycoside is a molecule formed by the bonding of a sugar to another group on its anomeric carbon via a glycosidic bond. Glycosides can be linked by O- (to form O-glycosides), N- (to form glycosylamines), S- (to form thioglycosides), or C- (to form C-glycosides) glycosidic bonds. Its experimental formula is C m(H 2O) n (where m can be different from n, m and n < 36). The glycosides in this invention include glucose (dextrose), fructose (levose), allose, arbutin, mannose, gulose, idose, galactose, tarose, galactosamine, glucosamine, sialic acid, N-acetylglucosamine, sulfoquinolate (6-deoxy-6-sulfo-D-guananoglucose), ribose, arabinose, xylose, lysose, sorbitol, mannitol, sucrose, lactose, maltose, trehalose, maltodextrin, raffinose, glucuronic acid (glucuronic acid), and stachyose. It can be in the configuration of D or L, in the form of a 5-atom cyclic furanose, in the form of a 6-atom cyclic guananose, or in an acyclic form, an α-isomer (where the -OH of the anomeric carbon is below the plane of the carbon atom in the Haworth projection), or a β-isomer (where the -OH of the anomeric carbon is above the plane of the carbon atom in the Haworth projection). In this invention, it is also referred to as a monosaccharide, disaccharide, polyol, or oligosaccharide containing 3-6 sugar units.

[0033] In this invention, "antibody" refers to a full-length immunoglobulin molecule or the immunoactive portion of a full-length immunoglobulin molecule, such as a molecule containing an antigen-binding site that specifically binds to a target antigen or a portion of a target antigen. Targets include, but are not limited to, cancer cells or cells that produce autoantibodies associated with autoimmune diseases. The immunoglobulins in this invention can be any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulins. Immunoglobulins can be derived from any species, but are preferably derived from humans, mice, or rabbits. The antibodies of the present invention are preferably monoclonal antibodies, including but not limited to polyclonal, monoclonal, bispecific, human, humanized or chimeric antibodies, single-chain antibodies, Fv, Fab fragments, F(ab') fragments, F(ab')2 fragments, fragments generated from Fab expression libraries, anti-idiotypic (anti-Id) antibodies, CDRs and any antigenic determinant binding fragments of the above structures that can immunely and specifically bind to cancer cell antigens, viral antigens or microbial antigens.

[0034] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a largely homogeneous population of antibodies, meaning that the individual antibodies constituting the population are identical, except for the possible presence of small amounts of naturally occurring mutations. Monoclonal antibodies exhibit high specificity against a single antigenic site. The modifier "monoclonal" indicates the characteristic of antibodies obtained from a largely homogeneous population of antibodies and should not be interpreted as requiring the production of antibodies by any particular method.

[0035] "Complete antibody" is an antibody that contains an antigen-binding variable region appropriate to its antibody class, as well as a light chain constant domain (CL) and heavy chain constant domains CH1, CH2, CH3, and CH4. The constant domain can be a natural sequence constant domain (e.g., a human natural sequence constant domain) or a variant of its amino acid sequence.

[0036] An "antibody fragment" is a part of a complete antibody, containing its antigen-binding region or variable region. Examples of antibody fragments include Fab, Fab', F(ab')sub2 and Fv fragments, biantibodies, triantibodies, tetraantibodies, linear antibodies, single-chain antibody molecules, scFv, scFv-Fc, multispecific antibody fragments formed from antibody fragments, fragments generated from Fab expression libraries, or antigenic determinant binding fragments of any of the above, which immune-specifically bind to target antigens (e.g., cancer cell antigens, viral antigens, or microbial antigens).

[0037] An "antigen" is an entity that an antibody specifically binds to.

[0038] The terms "specific binding" and "specifically binding" refer to the ability of an antibody or antibody derivative to bind to the corresponding antigenic determinant of its target antigen in a highly selective manner, without binding to a variety of other antigens. Typically, the antibody or antibody derivative binds to a specific antigen with an affinity of at least about 1 × 10⁻⁷ M, preferably 10⁻⁸ M to 10⁻⁹ M, 10⁻¹⁰ M, 10⁻¹¹ M, or 10⁻¹² M, which is at least twice the affinity for binding to nonspecific antigens other than the specific antigen or closely related antigens (e.g., BSA, casein).

[0039] A "mirror isomer," also known as an "optical isomer," is one of two stereoisomers that are mirror images of each other and cannot overlap (are not identical), much like a person's left and right hands. They cannot overlap unless flipped along a plane (changing their orientation alone cannot make them overlap). A single chiral atom or similar structural feature in a compound gives the compound two possible configurations that cannot overlap and are mirror images of each other. The presence of multiple chiral features in a compound increases the number of possible configurations, some of which may be mirror images of each other. An enantiomerically pure compound is a sample that possesses only the same chirality within the capability of a detection method. In a symmetrical environment, two enantiomers can cause plane-polarized light (+ / -) to rotate equally in opposite directions (polarized light can be considered an asymmetric mediator), and otherwise have the same chemical and physical properties. For this reason, they are sometimes called optical isomers. A mixture of an optically active isomer and its equal amounts of enantiomers is called a racemic mixture, which has no net rotation of plane-polarized light because each positive rotation (+) is completely canceled out by a negative rotation (-). Typically, two mirror-image isomers react differently with other mirror-image isomers. Since many biomolecules are enantiomers, the effects of two enantiomers on a biological organism can sometimes differ significantly. For example, in pharmaceuticals, usually only one enantiomer can exert the desired physiological effect, while the other enantiomer may have lower or no activity, or even produce adverse reactions. Based on this finding, drugs composed of only one enantiomer (“enantiomerically pure”) can be developed to enhance pharmacological potency and sometimes eliminate some side effects.

[0040] An isotope is a nuclide of a specific chemical element that has the same number of protons but different numbers of neutrons. All isotopes of the same element have the same number of protons. An atomic number designates a specific element, but is not an isotope; atoms of a specific element can have different numbers of neutrons. The number of nucleons (protons and neutrons) is the mass number of an atom, and each isotope of a specific element has a different mass number. For example, carbon-12, carbon-13, and carbon-14 are three isotopes of carbon, with mass numbers of 12, 13, and 14, respectively. Carbon has an atomic number of 6, meaning that each carbon atom has 6 protons, so these isotopes have 6, 7, and 8 neutrons, respectively. Hydrogen atoms have three isotopes: protium (¹H), deuterium (²H), and tritium (³H). Deuterium has twice the mass of protium, and tritium has three times the mass of protium. Through kinetic isotope effects, isotope substitution experiments can be used to determine the mechanisms of chemical reactions. Isotope substitution experiments can also be used to study how the body reacts to exogenous compounds after they enter the body through absorption and distribution mechanisms, metabolic changes in the body (e.g., through the action of metabolic enzymes such as cytochrome P450 or glucuronyl transferase), and the excretion pathways of drug metabolites; this is known as pharmacokinetic (PK) studies. Isotope substitution experiments can also be used to study the biochemical and physiological effects of drugs, including their effects on animals (including humans), microorganisms, or biological combinations (e.g., infections); this is known as pharmacodynamic (PD) studies. Both types of studies influence the dosage, benefits, and adverse effects of drugs. Isotopes can contain stable (non-radioactive) or unstable elements. Isotope-substituted drugs may have different therapeutic effects than the original drug. Isotope labeling of the compounds disclosed in this invention can be used for drug and / or individual tissue distribution determination. Tritium (3H) and carbon-14 (14C) labeled compounds are particularly preferred because they are easy to prepare and detect. Furthermore, substitution with heavier isotopes such as deuterium (2H) can provide certain therapeutic advantages due to greater metabolic stability, such as a longer in vivo half-life or reduced dose requirements, and may therefore be a preferred option in some cases. The isotopically labeled compounds disclosed in this invention, including their pharmaceutical salts, esters, and prodrugs, can be prepared by any method known in the art. The same benefits can also be obtained by using 12C instead of natural 12C.

[0041] "Pharmaceutical acceptable" or "pharmaceuticalally acceptable" means that when the molecular entity and composition are properly administered to animals or humans, they will not produce adverse, allergic, or other adverse reactions.

[0042] "Pharmaceutically acceptable solvate" or "solvent" means a combination of one or more solvent molecules with a compound disclosed in this invention. Examples of solvents used to prepare solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

[0043] "Pharmaceutically acceptable excipients" include any carriers, diluents, adjuvants, or others such as preservatives or antioxidants, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, solvents, dispersants, coating agents, antibacterial and antifungal agents, isotonic agents, and absorption delay agents. The use of these mediators and agents for the active pharmaceutical ingredient is well-known in the industry. Any conventional mediator or agent, unless incompatible with the active ingredient, may also be considered for use in therapeutic compositions. Supplemental active ingredients may also be incorporated into the composition to create a suitable therapeutic combination.

[0044] As used herein, "pharmaceutically acceptable salt" refers to a derivative of the compound disclosed in this invention, obtained by preparing an acid salt or base salt of the parent compound. Pharmaceutically acceptable salts include conventionally non-toxic salts or quaternary ammonium salts formed from a non-toxic inorganic or organic acid and the parent compound. For example, conventionally non-toxic salts include salts derived from inorganic acids (e.g., hydrochloric acid, hydrobromide, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, etc.); and salts prepared from organic acids (e.g., acetic acid, propionic acid, succinic acid, tartaric acid, citric acid, methanesulfonic acid, benzenesulfonic acid, glucuronic acid, glutamic acid, benzoic acid, salicylic acid, toluenesulfonic acid, oxalic acid, fumaric acid, maleic acid, and lactic acid, etc.). Other addition salts include ammonium salts, such as salts of trimethylamine, meglumine, glycerol, etc., and metal salts, such as sodium, potassium, calcium, zinc, or magnesium salts.

[0045] The pharmaceutically acceptable salts of this invention can be synthesized from a parent compound containing a basic or acidic moiety using conventional chemical methods. Generally, these salts can be prepared by adding an equimolar amount of a suitable base or acid to the parent compound in water or an organic solvent, or a mixture of both. Generally, diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred non-aqueous media. A list of suitable salts is available in Remington's Pharmaceutical Sciences, published by Mack Publishing in 1985, the contents of which are incorporated herein by reference.

[0046] The term "therapeutic effective dose" refers to the amount of a conjugate that is effective in treating a disease or symptom in mammals. In cancer treatment, a therapeutically effective dose of a conjugate can reduce the number of cancer cells; reduce the size of the tumor; inhibit (i.e., to some extent slow down, ideally stop) the infiltration of cancer cells into peripheral organs; inhibit (i.e., to some extent slow down, ideally stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent. In terms of the extent to which a drug can inhibit growth and / or kill existing cancer cells, it may be cytoseptic and / or cytotoxic. For cancer treatment, efficacy can be assessed, for example, by evaluating time to disease progression (TTP) and / or determining the response rate (RR).

[0047] As used herein, the term "pharmaceutically acceptable form" refers to the form of the disclosed compound, including but not limited to pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs, and isotopically labeled derivatives. In one embodiment, "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, prodrugs, and isotopically labeled derivatives. In some embodiments, "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable isomers and their stereoisomers, prodrugs, and isotopically labeled derivatives.

[0048] The term “substantial” or “essentially” means the majority of a mixture or sample, i.e., >50%, preferably more than 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0049] The combinations of substituents and variables contemplated in this invention refer only to those combinations that can form stable compounds. The term "stable" as used herein means stable during manufacturing and maintaining the integrity of the compound for the period of time used for the purposes detailed herein (e.g., therapeutic or preventative administration to an individual).

[0050] The compounds of the present invention are, after preparation, preferably isolated and purified to obtain a composition with an content equal to or greater than 95% (“pure”), which is then used or formulated as described herein. In some embodiments, the compounds of the present invention have a purity exceeding 99%.

[0051] The term "cytotoxic activity" refers to the cytotoxic effect of a drug, a camptothecin conjugate, or a camptothecin conjugate's intracellular metabolite. Cytotoxic activity can be expressed as the IC50 value, which is the concentration (moles or mass / unit volume) at which half of the cells survive.

[0052] The term "cell inhibitory activity" refers to the antiproliferative effect of a drug or camptothecin analogue conjugate or its intracellular metabolites.

[0053] As used herein, the term "cytotoxic agent" refers to a substance that has cytotoxic activity and causes cell destruction. This term is intended to include chemotherapeutic agents and toxins, such as small molecule toxins or enzyme-active toxins derived from bacteria, fungi, plants, or animals, including their synthetic analogues and derivatives.

[0054] "Administer" or "place" means to transfer, deliver, introduce, or transport a drug or other agent to an individual by any means. These means include oral, local contact, intravenous, intraperitoneal, intramuscular, intralesional, intranasal, subcutaneous, or intrathecal administration. The present invention also contemplates the use of devices or instruments for administering the agent. Such devices may be active or passive delivery methods and may be sustained-release or rapid-release delivery devices.

[0055] In the context of cancer, the term "treatment" includes any or all of the following: stopping the growth and replication of tumor cells or cancer cells, reducing the overall tumor mass, and improving one or more symptoms associated with the disease.

[0056] In the context of autoimmune diseases, the term "treatment" includes any or all of the following: stopping the replication of cells associated with the autoimmune disease, including but not limited to cells capable of producing autoimmune antibodies, reducing the amount of autoimmune antibodies, and improving one or more symptoms of the autoimmune disease.

[0057] In the context of infectious diseases, the term "treatment" includes any or all of the following: preventing the growth, proliferation, or replication of the pathogen that causes the infectious disease, and improving one or more symptoms of the infectious disease.

[0058] The terms “cancer” and “cancer-related” refer to or describe a physiological condition or symptom in mammals, typically characterized by unregulated cell growth. A “tumor” includes one or more cancer cells.

[0059] As used in this article, "autoimmune disease" refers to a disease or condition caused by and targeting an individual's own tissues or proteins.

[0060] As used herein, "patient" refers to an individual who has applied the camptothecin conjugate of the present invention. Patients include, but are not limited to, humans, rats, mice, guinea pigs, non-human primates, pigs, goats, cattle, horses, dogs, cats, birds, and poultry. Typically, patients are rats, mice, dogs, humans, or non-human primates, especially humans.

[0061] Examples of “mammals” or “animals” include, but are not limited to, humans, rats, house rats, guinea pigs, monkeys, pigs, goats, cows, horses, dogs, cats, birds, and poultry.

[0062] Unless the context otherwise requires, the term "treatment" or "curative therapy" refers to therapeutic and preventative treatment aimed at suppressing or slowing (alleviating) undesirable physiological changes or disorders, such as the development or spread of cancer. For the purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, relief of symptoms, reduction of disease severity, stable (i.e., non-deterioration) of the disease, delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can also refer to extended survival compared to expected survival without treatment. Those in need of treatment include those who already have the condition or disorder and those who are susceptible to it.

[0063] Drug-linker-binding ligand conjugates

[0064] As described above, the present invention provides conjugates of cell surface-binding molecules—camptothecin analogues—as shown in formula (I): (I) Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; Where T is the target or binding ligand; L is the releasable linker; It is a bonding bond where L is independently connected to the atoms R1, R2, R3, or R5 within the parentheses; n is 1-30, m is 1-10; The part in parentheses is an active camptothecin analogue, wherein: R1 and R2 are independently H; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy, alkylcarboxylic acid, or carbonyl; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently H, C(O)NH, C(O)O, SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, C(O)R6, C(O)NHR6; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy, alkylcarboxylic acid, or carbonyl; C2-C 6. Heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkylamide, aminoalkylamide, oxime; NH₂ or OH; R4 halogens (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NHR6, N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is H, C(O)O, C(O)NH, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether of C2-C6, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), or alkyl carboxylic acid; heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl C2-C6 amide or amino acid; or a pharmaceutically acceptable salt; Furthermore, R1, R2, R3, and R6 can be omitted independently, and together with R2, R3, X, C-10, and C-9, they can form 5-, 6-, or 7-membered heterocycles.

[0065] In another embodiment, the conjugate of camptothecin analogue has the structure of formula (II): (II) Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; Where T is the targeting or binding ligand; L is the releasable linker; n is 1-30, and m is 1-10; The part in parentheses is an effective camptothecin analogue, wherein: R1 is a straight-chain or branched C1-C6 alkyl, alkoxy, alkylamino (including first, second, third amino or quaternary ammonium), oxycarbonyl, aminocarbonyl, aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, oxycycloalkyl, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkylamide, aminoalkylamide, oxime; NH, or O; R 2 is H, a straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium), aminoalkyl alcohol, aminoalkylamine, oxyalkyl alcohol, oxyalkylamine, aminoalkyl, oxyalkyl, or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxime; NH 2 or OH; R3 is independently H; R6NHC(O), R6OC(O), SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, R6C(O), C(O)NR6R6'; linear or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl acetylamine, oxime; C5-C12 glycoside; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NHR6, N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R 5 is H, C(O)OR 6, C(O)NHR 6, R 6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide, or amino acid; or a pharmaceutically acceptable salt; In addition, R1 can be omitted and C-7 can be directly connected to L, and R2, R3, X, C-10 and C-9 can together form a 5-, 6- or 7-membered heterocyclic ring.

[0066] The exemplary compound within the parentheses of formula (II) has the following structure: II-01, II-02, II-03, II-04, II-05, II-06, II-07, II-08, II-09, II-10, II-11, II-12, II-13, II-14, II-15, II-16, II-17, II-18, II-19, II-20, II-21, II-22, II-23, II-24, II-25, II-26, II-27, II-28, II-29, II-30, II-31, II-32, II-33, II-34, II-35, II-36, II-37, II-38, II-39, II-40, II-41, II-42, II-43, II-44, II-45, II-46, II-47, II-48, II-49, II-50, II-51, II-52, II-53, II-54, II-55, II-56, II-57, II-58, II-60, II-61, Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; The definitions of R6 and R6' are the same as those in the previous text. in" “ is the connection point of connector L connected to equation (II).

[0067] In another specific embodiment, the conjugate of the cell-binding molecule-camptothecin analog has the structure of formula (III): (III) Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; Where T is the targeting or binding ligand; L is the releasable linker; n is 1-30, and m is 1-10; The part in parentheses is an active camptothecin analogue, in which: R 1 is a straight-chain or branched C1-C6 alkyl, alkoxy, alkylamino (including first, second, and third amino groups or quaternary ammonium groups), oxycarbonyl, aminocarbonyl, aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy, or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, oxycycloalkyl, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkyl ether, aminoalkyl ether, oxyalkyl ester, aminoalkyl ester, oxyalkylamide, aminoalkylamide, oxime; NH or O; R2 is NH, NR6, -N+R6R6'-, O, S, straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium), aminoalkyl alcohol, aminoalkylamine, oxyalkyl alcohol, oxyalkylamine, aminoalkyl, oxyalkyl or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxime; oxyalkyl ether, aminoalkyl ether, oxyalkyl ester, aminoalkyl ester, oxyalkylamide, aminoalkylamide; R3 is H, R6NHC(O), R6OC(O), SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, R6C(O), C(O)NR6R6'; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl acetylamine, oxime; C5-C12 glycoside; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NHR6, N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is H, C(O)OR6, C(O)NHR6, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide, or amino acid; or a pharmaceutically acceptable salt; Furthermore, R2 is independently defaulted and C-9 is directly connected to L, and R2, R3, X, C-10, and C-9 can together form 5-, 6-, or 7-membered heterocyclic rings.

[0068] The exemplary compound within the parentheses of formula (III) has the following structure: III-1, III-2, III-3, III-4, III-5, III-6, III-7, III-8, III-9, III-10, III-11, III-12, III-13, III-14 III-15 III-16 III-17, III-18, III-19, III-20, III-21, III-22, III-23, III-24, III-25, III-26, III-27, III-28, III-29, III-30, III-31, III-32, III-33, III-34, III-35, III-36, III-37, III-38, III-39, III-40, III-41, III-41, III-42, III-43, III-44, III-45, III-46, III-47, III-48, III-49, III-50, III-51, III-52, Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or the polycrystalline structure of these compounds; or their isomers, optical isomers, racemic mixtures, non-mirror isomers, or mirror isomers; R6, and R6' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide, or amino acid; or a pharmaceutically acceptable salt;

[0069] In another embodiment, the conjugate of the cell-binding molecule-camptothecin analog has formula (IV): (IV) Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; Where T is the target or binding ligand; L is the releasable linker; n is 1-30 and m is 1-10; The part in parentheses is an effective camptothecin analogue, wherein: R1 and R2 are independently H, NR6R6', -N+R6R6'R6'', OH, SH, C1-C6 straight-chain or branched alkyl, alkoxy, alkylamino (including first, second, third amino or quaternary ammonium), oxycarbonyl, aminocarbonyl, aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, oxycycloalkyl, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkyl ether, aminoalkyl ether, oxyalkyl ester, aminoalkyl ester, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently -NHC(O)-, -C(O)-, SO2-, -SO2NH-, -NR6SO2-, R6NHC(O), R6OC(O), SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, R6C(O), C(O)NR6R6'; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including primary, secondary, and tertiary amines) or alkyl carboxylic acid; C2-C 6. Heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxime; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NH(R6)S(O2)R6', N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is H, C(O)OR6, C(O)NHR6, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide, or amino acid; or a pharmaceutically acceptable salt; In another embodiment, R3 is independently defaulted and X of C-10 is directly connected to L, and R2, R3, X, C-10 and C-9 can together form a 5-, 6- or 7-membered heterocyclic ring.

[0070] The exemplary compound within parentheses of formula (IV) has the following structure: IV-1, IV-2, IV-3 IV-4, IV-5, IV-6, IV-7, IV-8, IV-9, IV-10, IV-11, IV-12, IV-13, IV-14, IV-15, IV-16, IV-17, IV-18, IV-19, IV-20, IV-21, IV-22, IV-23, IV-24, IV-25, IV-26, IV-27, IV-28, IV-29, IV-30, IV-31, IV-32, IV-33, IV-34, IV-35, IV-36, IV-37, IV-38, IV-39, IV-40, IV-41, IV-42, IV-43, IV-44, IV-45, IV-46, IV-47, Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; wherein “ The definitions of R6 and R6' are as described above;

[0071] In another embodiment, the conjugate of the cell-binding molecule-camptothecin analog has the structure of formula (V): (V) Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; Where T is the target or binding ligand; L is the releasable linker; n is 1-30, m is 1-10; The part in parentheses is an active camptothecin analogue, wherein: R1 and R2 are independently H, NR6R6', -N+R6R6'R6'', OH, SH, straight-chain or branched C1-C6 alkyl, alkoxy, alkylamino (including first, second, third amino or quaternary ammonium), oxycarbonyl, aminocarbonyl, aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, oxycycloalkyl, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkyl ether, aminoalkyl ether, oxyalkyl ester, aminoalkyl ester, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently R6NHC(O)-, R6C(O)-, R6SO2, -SO2NHR6, R6OC(O), R6'SO2R6-, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, R6C(O), C(O)NR6R6'; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including primary, secondary, and tertiary amines) or alkyl carboxylic acid; C2-C 6. Heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxime; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NH(R6)S(O2)R6', N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is C(O)O, C(O)NH, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide, or amino acid; or a pharmaceutically acceptable salt; In addition, R5 is independently defaulted and the O of C-20 is directly connected to L, and R2, R3, X, C-10 and C-9 can together form 5-, 6- or 7-membered heterocycles.

[0072] The exemplary compound within the parentheses of formula (V) has the following structure: V-01, V-02, V-03, V-04, V-05, V-06, V-07, V-08, V-09, V-10, V-11, V-12, V-13, V-14, V-15, V-16, V-17, V-18, V-19, V-20, V-21, V-22, V-23, V-24, V-25, V-26, V-27, V-28, V-29、 V-30、 V-31、 V-32、 V-33、 V-34、 V-35、 V-36、 V-37、 V-38、 V-39、 V-40、 V-41、 V-42、 V-43、 V-44、 V-45、 V-46、 V-47、 V-48、 V-49、 V-50、 V-51、 V-52, V-53, V-54 V-55, V-56 V-57 V-58 V-59 V-60 V-61, Or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of these compounds; or their isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers; wherein “ R6 and R6' are as defined above; In another embodiment, the route for synthesizing camptothecin analogues and their conjugates with the cell surface receptor binding molecules of the present invention is included, but is not limited to, those shown in Figures 1-32.

[0073] In another embodiment, the releasable linker (L) is a chain-like structure composed of atoms selected from the group consisting of C, N, O, S, Si, and P, covalently linking a cell surface-bound ligand (T) to an effective camptothecin analog. The linker can have various lengths, for example, in the range of about 2 to about 100 atoms. The atoms used to form the linker can be combined in any chemical manner, for example forming alkyl, alkenyl and alkyneyl groups, ethers, polyoxyethylene, esters, amines, imines, polyamines, hydrazines, amides, ureas, aminoureas, carbapenems, alkoxyamines, carbapenems, alkoxyamines, carbamates, amino acids, acetoamines, isohydroxamic acids, etc. Furthermore, it should be noted that the atoms forming the releasable linker (L) can be saturated or unsaturated, or can be free radicals, or can be cyclic to form a divalent cyclic structure, including cycloalkanes, cyclic ethers, cyclic amines, aryl groups, heteroaryl groups, etc.

[0074] The term "releasable linker" refers to a linker containing at least one bond that can be broken under physiological conditions, such as pH, acidity, alkalinity, oxidation, metabolism, biochemistry, or enzymatic instability. It should be understood that bond breaking is not necessarily a biological or metabolic process, but may be a chemical reaction, such as hydrolysis or substitution. Examples of such physiological conditions include: endosomes with a pH lower than the intracellular pH; and malignant cell memory cells containing large concentrations of glutathione, which can undergo disulfide exchange reactions with intracellular thiols.

[0075] The releasable linker L of the coupling can have the following structure: --Ww-(Aa)r-Vv--; where: --W-- is an extension unit; w is 0 or 1; each -Aa- is independently an amino acid unit; r is independently an integer from 0 to 12; --V-- is a spacer unit; v is 0, 1 or 2.

[0076] When the extended body –Ww – is present, it serves to connect the cell-binding molecule (T) and the amino acid unit –Aa – or the spacer –V – (when –Aa – is absent). The extended body W can independently contain a self-destructing spacer, peptide unit, hydrazone bond, disulfide bond, or thioether bond. In this case, the cell-binding molecule (T) contains a group capable of bonding with the corresponding functional group on the extended body. Functional groups inherent to the cell-binding molecule or generated chemically for connection include, but are not limited to: thiol (--SH), amino group, hydroxyl group, carbonyl group, anomeric hydroxyl group of sugar, and carboxyl group. Preferred functional groups are thiol, carboxyl, and amino groups. Thiol groups can be generated by reducing the disulfide bonds within the cell-binding molecule. Thiol groups can also be generated by reacting the amino group of lysine in cell-binding molecules with 2-iminothione (Traut reagent), thiolactone, or other thiol-generating reagents. For example, cell-binding molecule T can be modified with a linker or thioester containing a disulfide bond, and then thiol groups can be generated by reduction or hydrolysis, respectively.

[0077] An example of W connected to T is shown below: ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; , , , , , , , , , , , , , , , , , , , , , R20 and R21 are selected from the following groups of composition: –C1-C9 alkyl-, -C1-C7 carbon ring-, -O-(C1-C8 alkyl)-, -arylene-, -C1-C9 alkyl-arylene-, -arylene, -C1-C9 alkyl-, -C1-C9 alkyl-(C1-C8 carbon ring)-, -(C3-C7 carbon ring)-C1-C9 alkyl-, -C3-C8 heterocyclic, -C1-C10 alkyl-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C9 alkyl-, -(CH2CH2O)k-, -(CH(CH3)CH2O)k- and -(CH2CH2O)k-CH 2-; k is an integer from 1 to 20; R' and R'' are independently H or CH 3.

[0078] In another embodiment, the covalent connection of W and T as described above can be achieved through various chemical reactions.

[0079] Examples of amide bond formation are as follows: The extended unit contains a reactive site E, which can react with the first or second amine on the cell-binding molecule to form an amide bond. Examples of E include, but are not limited to, hydroxysuccinimide esters (NHS and S-NHS, etc.), 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters (including S-tetrafluorophenyl esters), acid anhydrides, amide chlorides, sulfonamide chlorides, isocyanates, and isothiocyanates.

[0080] Examples of linkages using sulfides or disulfides: The extended unit contains a site that can react with a thiol group, and can form a thioether or disulfide bond with the cell-binding molecule T. To form a disulfide bond, the thiol group on the cell-binding molecule T can be generated either by reducing the intramolecular disulfide bond or by modifying the cell-binding molecule T through other chemical processes.

[0081] In another embodiment of the invention, the extension may also have an active group capable of reacting with an aldehyde group (-CHO) or a ketone group (-C(=O)R) to modify the aldehyde or ketone group on the cell-binding molecule T. For example, the sugar in the cell-binding molecule T can be oxidized with an oxidizing agent such as sodium periodate to generate an aldehyde or ketone group (-C(=O)R). Furthermore, the amine on the N-terminal amino acid of an antibody (or protein or polypeptide) can react with pyridoxal 5-phosphate (PLP) to introduce a ketone group (Scheck & Francis, ACS Chem. Biol. 2007, 2, 247-251). These aldehyde or ketone groups (-C=O) can react with active reactive groups on the extension, such as acehydrazide, oxime, a first or second amine, hydrazine, thiourea, hydrazide carboxylate, or arylhydrazine, thereby linking the two together.

[0082] Examples of linkages using hydrazones, oximes, or imines are as follows: R20 and R21 are defined as described above, and R25 refers to a substituent of an amino acid.

[0083] In another embodiment of the invention, the extension (which may contain an insulator V and / or an amino acid) can first be linked to a cell-binding molecule (T), and then this cell-binding molecule-extension fragment can be conjugated with a camptothecin analog. This conjugation process can be carried out in a buffered aqueous solution. Below are some examples of two-step coupling in this manner (drugs linked to R 16 are omitted): E includes, but is not limited to, hydroxysuccinimide esters (NHS and S-NHS, etc.), 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters (including S-tetrafluorophenyl esters), acid anhydrides, acetyl chloride, sulfonyl chloride, isocyanates, and isothiocyanates, etc. R' and R'' are H or CH3, respectively; R20, R16, and Ar are defined as above; R26 is H, F, or NO2; J is F, Cl, Br, I, toluenesulfonate group (TsO), or methanesulfonate group (MsO), wherein... Contains at least one camptothecin analogue, such as As shown.

[0084] In another embodiment of the invention, the extension can be first linked to a camptothecin analogue, and then coupled to a cell-binding molecule (T) in a buffered aqueous solution (containing up to 50% organic solvent) at a pH of 3-10 (preferably 5.0-8.5). An example of this two-step coupling is as follows: E includes, but is not limited to, hydroxysuccinimide esters (NHS and S-NHS, etc.), 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters (including S-tetrafluorophenyl esters), acid anhydrides, acetyl chloride, sulfonyl chloride, isocyanates, and isothiocyanates, etc. R' and R'' are H or CH3, respectively; R20, R16, and Ar are defined as above; R26 is H, F, or NO2; J is F, Cl, Br, I, toluenesulfonate group (TsO), or methanesulfonate group (MsO), wherein... It contains at least one camptothecin analogue.

[0085] When the amino acid unit (--Aa--) is present, it connects the extended body and the isolated body when the isolated body is present; when the isolated body is absent, it connects the extended body and the camptothecin analog; if both the extended body and the isolated body are absent, it directly connects the cell-binding molecule T and the camptothecin analog. --(Aa)r- can be a natural or non-natural amino acid, and can be a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide unit, where r is an integer between 0 and 12. Here, amino acid generally refers to aminoalkyl carboxylic acid, where the alkyl group can be an alkyl group substituted with groups such as alkyl, acetyl, hydroxyalkyl, mercaptoalkyl, aminoalkyl, or carboxylalkyl. The structures of natural and non-natural amino acids and peptides are detailed in *Amino Acid and Peptide* by GC Barrett and DT Elmore (Cambridge University Press, 2004). In addition, amino acids also refer to β-amino acids, γ-amino acids, and long-chain amino acids containing methyl, benzyl, hydroxymethyl, mercaptomethyl, carboxyl, carboxymethyl, or guanidinopropyl groups within their molecules. Preferred amino acids include arginine, aspartic acid, aspartic acid, citrulline, cysteine, glycine, glutamic acid, leucine, lysine, glutamine, serine, ornithine, phenylalanine, threonine, tyrosine, and valine.

[0086] The amino acid unit can be cleaved by one or more enzymes, including tumor-associated proteolytic enzymes, to release a camptothecin analog. In one embodiment, the camptothecin analog is protonated in vivo after release.

[0087] When the insulator (-V-) is present, it links the amino acid unit to the camptothecin analogue; when the amino acid unit is missing, it links the extension unit to the camptothecin analogue. When both the amino acid unit and the extension unit are missing, the insulator links the camptothecin analogue to the cell-binding molecule (T). The insulator may contain functional groups to improve the water solubility, biotransport, and optimal renal clearance, uptake, absorption, biodistribution, and bioavailability of the conjugate. There are two basic types of insulators: self-destructive and non-self-destructive. Non-self-destructive insulators refer to those where the amino acid unit of the camptothecin analogue-linker-cell-binding molecule or camptothecin analogue-linker is broken (especially after enzymatic hydrolysis), with some or all of the insulator remaining attached to the camptothecin analogue. Self-destructive compounds include aromatic compounds with electronic structures similar to p-aminobenzylaminomethyl (PAB), 2-aminoimidazolium-5-methanol derivatives, heterocyclic PAB derivatives, β-glucuronic acid, ortho- or p-aminobenzyl acetals, or one of the following structures: , , , . , (*) indicates an insulator, a releasable linker unit, an amino acid (Aa)r, a camptothecin analog, or a cell-binding molecule (T) linker; X, Y, and Z3 are independently NH, O, or S; Z2 is independently H, NH, O, or S; v is 0 or 1; Q is independently H, OH, C1-C6 alkyl, (OCH2CH2)n, F, Cl, Br, I, OR17, or SR17, NR17R18, N=NR17, N=R17, NR17R18, NO2, SOR17R18, SO2R17, SO3R17, OSO3R17, PR17R18, POR17R18, PO2R17R18, OPO(OR17)(OR18) 18), or OCH 2PO(OR 17)(OR 18), wherein R 17 and R 18 are independently H, C 1-C 8 alkyl, C 2-C 8 alkenyl, alkyne, heteroalkyl, C 3-C 8 aryl, heterocyclic, carbocyclic, cycloalkyl, heterocyclic, heteroaryl, alkylcarbonyl, or pharmaceutically acceptable ionic salt; v is an integer from 1 to 20.

[0088] Examples of non-self-destructive intervals (-V-): ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 6-Maliimidohexyl (MC) Maleiminopropyl (MP) val-citrulline (val-cit) Alanine-phenylalanine (ala-phe) Lysine-phenylalanine (lys-phe) p-Aminobenzyloxy-carbonyl (PAB) 4-Thiopentanoic acid (SPP) 4-Thiobutyryl (SPDB), 4-(N-maleiminomethyl)cyclohexane-1-acryl (MCC) Maleimide ethyl (ME) 4-Thio-2-hydroxysulfonyl-butyryl (2-Sulfo-SPDB), Arylthiols (PySS) (4-Acetyl)aminobenzoyl (SIAB) ,oxybenzyl thio, Aminobenzylthio, Dioxybenzyl thio, Diaminobenzylthio, Aminooxybenzyl sulfide group, Alkoxyamine (AOA) Ethylene oxide (EO), 4-Methyl-4-dithiovalerate (MPDP) Triazole, disulfide, Alkyl sulfonyl, Alkylsulfonamide, sulfonamide, Phosphine diamide, Alkylphosphonamide, Phosphine, N-methylphosphonoamine, N,N'-Dimethylphosphonoamine, N,N'-Dimethylphosphine diamide, hydrazine, Ethamidin; Oxime, Diacetylhydrazine, aminoethylamine, Aminoethyl-aminoethylamine, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or L- or D-, or natural or non-natural peptides containing 1-20 amino acids; Among them, "*" and " " is a spacer or releasable linker, a linking point for camptothecin analogs and / or bound molecules; m is 1-10; n is 1-20; X2, X3, X4, X5 or X6 are independently selected from the following groups of composition: NH, NHNH, N(R12), N(R12)N(R12'), O, S, C1-C6 alkyl, C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; CH2OR12, CH2SR12, CH2NHR12 or 1-8 amino acids; wherein R12 and R12' are independently H, C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, ester, ether or amide with 1-8 carbon atoms; or polyethylene oxide units of the formula (OCH 2CH 2) p or (OCH 2CH(CH 3)) p, wherein p is an integer from 0 to about 1000, or combinations thereof.

[0089] The breakable linker L contains at least one bond that is broken under physiological conditions or is unstable under pH, acid, base, oxidation, metabolic, biochemical, or enzymatic conditions, and has one of the following structures: -(CR15R16)m(Aa)r(CR17R18)n(OCH2CH2)t-, -(CR15R16)m(CR17R18)n(Aa)r(OCH2CH2)t-, -(Aa)r-(CR15R16)m(CR17R18)n(OCH2CH2)t-, -(CR15R16)m(CR17R18)n(OCH2CH2)r(Aa)t-, -(CR15R16)m-(CR17=CR18)(CR19R20)n(Aa)t(OCH2CH2)r-, -(CR15R16)m(NR11CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m(Aa)t(NR21CO)(CR19R20)n(OCH2CH2)r-, -(CR15R16)m(OCO)(Aa)t(CR19R20)n(OCH2-CH2)r-, -(CR15R16)m(OCNR17)(Aa)t(CR19R20)n(OCH2CH2)r-, -(CR15R16)m-(CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m(NR21CO)(Aa)t(CR19R20)n(OCH2CH2)r-, -(CR15R16)m(OCO)(Aa)t-(CR19R20)n(OCH2CH2)r-, -(CR15R16)m(OCNR17)(Aa)t(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m-(CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m-phenyl-CO(Aa)t-(CR17R18)n-, -(CR15R16)m-furan-CO(Aa)t(CR17R18)n-, -(CR15R6)m-oxazole-CO(Aa)t(CR17R18)n-, -(CR15R16)mthiazolylCO-(Aa)t(CCR17R18)n-, -(CR15R16)t-thiophene-CO(CR17R18)n-,-(CR 15R 16) t-imidazolium-CO-(CR 17R 18) n-, -(CR 15R 16) t-morpholine-CO(Aa) t(CR 17R 18) n-, -(CR 15R 16) t-piperazine-CO(Aa) t(CR 17R 18) n-, -(CR 15R 16) tN-methylpiperazine-CO(Aa) t(CR 17R 18) n-, -(CR 15R 16) m-(Aa) tphenyl-, -(CR 15R 16) m-(Aa) tfuran-, -(CR 15R 16) m-oxazole(Aa) t-, -(CR 15R 16) m-thiazolyl(Aa) t-, -(CR 15R 16) m-thiophene-(Aa) t-, -(CR 15R 16) m-imidazolium(Aa) t-, -(CR 15R 16) m-morpholine-(Aa) t-, -(CR 15R 16) m-piperazine-(Aa) t-, -(CR 15R 16) mN-methylpiperazine-(Aa) t-, -K(CR 15R 16) m(Aa)r(CR 17R 18) n(OCH 2CH 2) t-, -K-(CR 15R 16) m(CR 17R 18) n(Aa) r(OCH 2CH 2) t-, -K(Aa) r-(CR 15R 16) m(CR 17R 18) n(OCH 2CH 2) t-, ‑K-(CR 15R 16) m(CR 17R 18) n(OCH 2CH 2) r(Aa) t-, ‑K(CR 15R 16) m(CR 17=CR 18)(CR 19R 20) n(Aa) t(OCH 2-CH 2) r, ‑K(CR 15R 16) m(NR 11CO)(Aa) t-(CR 19R 20) n(OCH 2CH 2) r-, ‑K(CR 5R 6) m(Aa) t(NR 21CO)-(CR 19R 20) n(OCH 2CH 2) r-, ‑K(CR 15R 16) m-(OCO)(Aa) t(CR 19R 20) n-(OCH 2CH 2) r-, ‑K(CR 15R 16) m(O-CNR 17)(Aa) t(CR 19R 20) n-(OCH 2CH 2) r-, ‑K(CR 15R 16) m(CO)(Aa) t-(CR 19R 20) n(OCH 2CH 2) r-,‑K(CR 15R 16) m(NR 21CO)(Aa) t(CR 19R 20) n(OCH 2CH 2) r-, ‑K(CR 15R 16) m-(OCO)(Aa) t(CR 19R 20) n(O-CH 2CH 2) r-, ‑K(CR 15R 16) m(OCNR 17)(Aa) t(CR 19R 20) n(OCH 2CH 2) r-, ‑K-(CR 15R 16) m(CO)(Aa) t-(CR 19R 20) n(OCH 2CH 2) r-, ‑K(CR 15R 16) m-phenyl‑CO(Aa) t(CR 17R 18) n-, -K-(CR 15R 16) m-furan-CO-(Aa) t(CR 17R 18) n-, -K(CR 15R 16) m-oxazol-CO(Aa) t(CR 17R 18) n-, -K(CR 15R 16) m-thiophene-CO(Aa) t-(CR 17R 18) n-, -K(CR 15R 16) t-thiophene-CO(CR 17R 18) n-, -K(CR 15R 16) timidazol-CO-(CR 17R 18) n-, -K-(CR 5R 6) toxazine-CO(Aa) t-(CR 17R 18) n-, -K(CR 15R 16) t-piperazine-CO(Aa) t(CR 17R 18) n-, ―K-(CR 15R 16) tN-methylpiperazine-CO(Aa) t(CR 17R 18) n-, ―K(CR 15R 16) m(Aa) tphenyl, ―K(CR 15R 16) m-(Aa) tfuran-, -K(CR 15R 16) m-oxazol-(Aa) t-, ―K(CR 15R 16) m-thiophene(Aa) t-, ―K(CR 15R 16) m-thiophene-(Aa) t-, ―K(CR 15R 16) m-imidazol(Aa) t-, ―K(CR 15R 16) m-oxazine(Aa) t-, ―K(CR 15R 16) m-piperazine(Aa) tG, -K(CR 5R 6)mN-methyl-piperazine(Aa) t-; where Aa, m, n, are defined above; t and r are independently 0–100; R 13, R 14, R 15, R 16, R 17, R 18, R 19, R 20, R 21 are independently selected from the following groups H: halogens; C1~C8 alkyl or heteroalkyl groups.C2~C8 aryl, alkenyl, alkynyl, ether, ester, amine, or amide; C3~C8 aryl, these groups can be replaced by one or more halogens, CN, NR 12R 12', CF 3, OR 12, aryl, heterocyclic, S(O)R 12, SO 2R 12, -CO 2H, -SO 3H, -OR 12, -CO 2R 12, -CONR 12, -PO 2R 12R 13, -PO 3H or P(O)R 12R 12'R 13; K is NH, NR 12, -SS-, -C(=O)-, -C(=O)NH-, -C(=O)O-, -C=NH-O-, -C=N-NH-, -C(=O)NH-NH-, O, S, Se, B, Het (a heterocyclic or heteroaromatic ring with C3-C12); or a peptide containing 1-20 amino acids, either the same or different.

[0090] In another embodiment of the invention, linker L preferably contains an amino group, a sulfonylurea, a phosphatidylamine, or an amino acid group, wherein formula (Iq) can be attached to linker L as a branch. The amino acid in linker L is preferably free from the group consisting of aspartic acid, glutamic acid, lysine, ornithine, or tyrosine, wherein one or both of the functional groups such as amino, carboxyl, or phenoxy groups can be attached to a long branch as shown in formula (Iq): (Iq); in This is the site that connects to a sulfonylurea, phosphate, amino, or carbonyl group in the linker, preferably to the carbonyl, amino, or phenoxy group of aspartic acid, glutamic acid, lysine, ornithine, or tyrosine; G1 is NH, NHNH, C(=O), NHNHC(O), C(=O)NH, C(=NH)NH, CH2, CH2C(O), C(O)O, NHC(O)NH or (Aa)r, (r = 1-12); G2 is NH, NHNH, C(=O), NHNHC(O), C(=O)NH, C(=NH)NH, CH2, C(O)O, NHC(O)NH, O, S, B, P(O)(OH), NHP(O)(OH), NHP(O)(OH)NH, CH2P(O)(OH)NH, OP(O)(OH)O, CH2P(O)(OH)O, NHS(O) 2. NHS(O)₂NH, CH₂S(O)₂NH, OS(O)₂O, CH₂S(O)₂O, Ar, ArCH₂, ArO, ArNH, ArS, ArNR₁, (Aa)r, (r = 1-12); X₁ and X₂ are independently O, CH₂, S, NH, N(R₁₂), +NH(R₁₂), +N(R₁₂)(R₁₃), C(O), OC(O), OC(O)O, NHSO₂NH, NHP(O)(NH)₂, SO₂NH, P(O)(NH)₂, NHS(O)NH, NHP(O)(OH)(NH), OC(O)NH, NHC(O)NH; Y₂ is O, NH, NR₁, CH₂S, Ar;G 3 is OH, SH, OR 1, SR 1, OC(O)R 1, NHC(O)R 12, C(O)R 12, CH 3, NH 2, NR 12, +NH(R 12), +N(R 12)(R 13), C(O)OH, C(O)NH 2, NHC(O)NH 2, BH 2, BR 12R 13. P(O)(OH) 2. NHP(O)(OH) 2. NHP(O)(NH 2) 2. S(O) 2(OH), (CH 2) q1C(O)OH, (CH 2) q1P(O)(OH) 2. C(O)(CH 2) q1C(O)OH, OC(O)(CH 2) q1C(O)OH, NHC(O)(CH 2) q1C(O)OH, CO(CH 2) q1P(O)(OH) 2. NHC(O)O(CH 2) q1-C(O)OH, OC(O)NH-(CH 2) q1C(O)OH, NHCO(CH 2) q1P(O)(OH) 2. NHC(O)(NH)(CH 2) q1C(O)OH, CONH(CH 2) q1P(O)(OH) 2, NHS(O) 2(CH 2) q1C(O)OH, CO(CH 2) q1S(O) 2(OH), NHS(O) 2NH-(CH 2) q1C(O)OH, OS(O) 2NH(CH 2) q1C(O)OH, NHCO(CH 2) q1S(O) 2(OH), NHP(O)(OH)(NH)-(CH 2) q1C(O)OH, CONH(CH 2) q1S(O)(OH), OP(O)(OH) 2, (CH 2) q1P(O)(NH) 2, NHS(O) 2(OH), NHS(O) 2NH 2, CH 2S(O) 2NH 2, OS(O) 2OH, OS(O) 2OR 1, CH 2S(O) 2OR 1, Ar, ArR 12, ArOH, ArNH 2, ArSH, ArNHR 12, or (Aa) q1; p 1, p 2 and p 3 are independently 0 -30 but not simultaneously 0; q 1 and q 2 are independently 0 -24;Preferably, G3 is a linear or branched C2-C50 polycarboxylic acid or C2-C50 polyalkylamine, C6-C90 oligosaccharide or polysaccharide, C6-C50 zwitterion containing quaternary ammonium cations and carboxylic acid anions or zwitterion containing quaternary ammonium cations and sulfonic acid anions (PSB), or a biodegradable polymer such as polylactic acid / glycolic acid (PLGA), poly(acrylate), deacetylated chitosan, copolymer of N-(2-hydroxypropyl)methacrylamide, poly[2-(methacryloxy)ethylphosphocholine] (PMPC), poly-L-glutamic acid, poly(lactide-co-glycolic acid) (PLG), poly(ethylene glycol) (PEG), or poly(propylene glycol) (PPG). Poly(ethylene glycol) modified peptides, poly(ethylene glycol) modified liposomes, poly(ethylene glycol) modified alkyl carboxylic acids, poly(ethylene glycol) modified alkylamines, hyaluronic acid (HA) (glycosaminoglycans), heparin or acetylheparin sulfate (HSGAG), chondroitin sulfate or ceremol sulfate (CSGAG), poly(ethylene glycol) modified alkyl sulfates, poly(ethylene glycol) modified alkyl phosphates, or poly(ethylene glycol) modified alkyl quaternary ammonium salts.

[0091] More preferably, formula (Iq) is selected from the following groups: (I-q01), (I-q02), (I-q03), (I-q04), (I-q04), (Iq-05), (I-q06), (I-q07), (I-q08) (Iq-09), (Iq-10), (Iq-11), (Iq-12), (Iq-13), (Iq-14), Among them, G1, p1, p2, p3, Aa, r, X2, q1, and m1 are the same as those defined above.

[0092] Alternatively, the binding molecule (T) of the present invention can be any kind of molecule that is currently known or forthcoming, capable of binding, compounding, or reacting with portions of cells that are therapeutically significant or can be biologically modified. The function of the binding molecule is to deliver camptothecin analogues to specific target cell populations that react with the binding molecule (T).

[0093] In another embodiment of the invention, the conjugates of CPT derivatives and cell-binding molecules have the following structures: (IIq-1), (IIq-2), (IIq-3), (IIq-4), (IIq-5), (IIq-6), (IIq-7), (IIq-8). (IIq-1), (IIq-2), (IIq-3), (IIq-4), (IIq-5), (IIq-6), (IIq-7), (IIq-8), Where R' and R'' are independently H, Me, Et, iPr, iBu, Bz (CH 2C 6H 5), CH 2COOH, CH 2CH 2COOH, CH 2CONH 2, CH 2CH 2CONH 2, CH 2CH 2CH 2NH 2, CH 2CH 2SCH 3, CH 2OH, CH 2CH 2CH 2NHC(=NH)NH 2, CH(OH)CH 3, CH 2C 6H 4OH, CH 2C 3N 2H 3; p 1 and p 2 are independently 0 ~ 24; q 1 is 1 ~ 18; q 3 is 0 ~ 6; q 4 is 0 ~ 4; m' and m'' are independently 0 ~ 6; m''' is 0 or 1; mAb is a cell-binding molecule, preferably an antibody; NH-drugs are compounds II-1~II-61, III-1~III-51, IV-1~IV-47 and V-1~V-61 listed above; n is 1~20; It is the site that is linked to NH-Drug.

[0094] Conjugates of formulas (IIq-1), (IIq-2), (IIq-3), (IIq-4), (IIq-5), (IIq-6), (IIq-7), and (IIq-8) are prepared by reacting the following formulas (IIq-9), (IIq-10), (IIq-11), (IIq-12), (IIq-13), (IIq-14), (IIq-15), and (IIq-16) with thiol-containing cell-binding molecules: (IIq-9), (IIq-10), (IIq-11), (IIq-12), (IIq-13), (IIq-14), (IIq-15), (IIq-8), Where R' and R'' are independently H, Me, Et, iPr, iBu, Bz (CH 2C 6H 5), CH 2COOH, CH 2CH 2COOH, CH 2CONH 2, CH 2CH 2CONH 2, CH 2CH 2CH 2CH 2NH 2, CH 2CH 2SCH 3, CH 2OH, CH 2CH 2CH 2NHC(=NH)NH 2, CH(OH)CH 3, CH 2C 6H 4OH, CH 2C 3N 2H 3; p 1 and p 2 are independently 0 ~ 24; q 1 is 1 ~ 18; q 3 is 0 ~ 6; q 4 is 0 ~ 4; m' and m'' are independently 0 ~ 6; m''' is 0 or 1; NH-Drug are compounds II-1 ~ II-61 listed above. III-1 ~ III-51, IV-1 ~ IV-47, and V-1 ~ V-61; It is the site that is linked to NH-Drug.

[0095] Free sulfide groups in proteins, particularly antibodies, can be generated by reducing interchain disulfide atoms. Reducing agents selected from the group consisting of dithiothreitol (DTT), dithioerythritol (DTE), L-glutathione (GSH), tris(2-carboxyethyl)phosphine (TCEP), 2-mercaptoethylamine (β-MEA), and / or β-mercaptoethanol (β-ME, 2-ME), are used in a buffer solution at pH 5.0–8.5. The reduction of two or more disulfide bonds in a protein with TCEP can be carried out before or simultaneously with coupling reactions of formulas (IIq-9), (IIq-10), (IIq-11), (IIq-12), (IIq-13), (IIq-14), (IIq-15), or (IIq-16). Sulfide groups can also be generated by reacting amino groups (lysine residues) on proteins with Traut's reagent (2-iminothione) or γ-thiobutyrolactone. The reaction of proteins with Traut reagent (2-iminothiacyclopentane) or γ-thiobutyrolactone compounds can be carried out in a one-pot (simultaneously) coupling reaction with compounds of the same formula (IIq-9), (IIq-10), (IIq-11), (IIq-12), (IIq-13), (IIq-14), (IIq-15), or (IIq-16).

[0096] Cell-binding agents (T) include, but are not limited to, high molecular weight proteins, such as intact antibodies (polyclonal antibodies, monoclonal antibodies); single-chain antibodies; antibody fragments such as Fab, Fab', F(ab')2, Fv (Parham, J. Immunol. 1983, 131, 2895-2902), fragments generated from Fab expression libraries, anti-idiotypic (anti-Id) antibodies; CDRs; antigenic determinant binding fragments of any of the above antibodies, capable of immune-specific binding to cancer cell antigens, viral antigens, microbial antigens; interferons (such as types I, II, and III); polypeptides; lymphocyte factors such as IL-2, IL-3, IL-4, and IL-6. GM-CSF, interferon-γ (IFN-γ); hormones such as insulin, TRH (thyrotropin-releasing hormone), MSH (melanocyte-stimulating hormone), steroid hormones such as androgens and estrogens; growth factors and colony-stimulating factors, such as epidermal growth factor (EGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), transforming growth factor (TGF), such as TGFα, TGFβ, insulin and insulin-like growth factor (IGF-I, IGF-II), G-CSF, M-CSF and GM-CSF (Burgess, Immunology Today 1984, 5, 155-158); Vaccinium growth factor (VGF); fibroblast growth factor (FGF); small molecular weight proteins; polypeptides; peptides and peptide hormones, such as bufotalin, gastrin, gastrin-releasing peptide; platelet-derived growth factors; interleukins and cytokines, such as interleukin-2 (IL-2), interleukin-6 (IL-6), leukemia suppressor factor, granulocyte-macrophage colony-stimulating factor (GM-CSF); vitamins, such as folic acid; apoproteins and glycoproteins, such as transferrin (O'Keefe et al., J. Biol. Chem. 1985 260 932-937); glycoproteins or lipoproteins, such as lectins; cellular nutrient transport molecules; small molecule inhibitors, such as prostate-specific membrane antigen (PSMA) inhibitors and small molecule tyrosine kinase inhibitors (TKIs); non-peptide or any other cell-binding molecules or substances, such as bioactive polymers (Dhar et al., Proc. Natl.). Acad. Sci. 2008, 105, 17356-61), bioactive dendritic polymers (Lee, et al., Nat. Biotechnol. 2005, 23, 1517-26; Almutairi, et al.; Proc. Natl. Acad. Sci. 2009, 106, 685-90), nanoparticles (Liong, et al., ACS Nano, 2008, 19, 1309-12; Medarova, et al., Nat .... Sci. 2009, 106, 685-90), nanoparticles (Liong, et al., ACS Nano, 2008, 19, 1309-12; Medarova, et al., Nat. Sci. 2008, 19, 1309-12; Medarova, et al., Nat. Sci. 2008, 105, 17356Med. 2007, 13, 372-7; Javier, et al., Bioconjugate Chem. 2008, 19, 1309-12), liposomes (Medinai, et al., Curr. Phar. Des. 2004, 10, 2981-9), and viral capsids (Flenniken, et al., Viruses Nanotechnol. 2009, 327, 71-93). Generally, if a suitable monoclonal antibody is available, the preferred monoclonal antibody is used as a cell surface binding agent.

[0097] Preferably, T is selected from the group of antibodies consisting of: single-chain antibodies, antibody fragments that bind to target cells, monoclonal antibodies, single-chain monoclonal antibodies, or monoclonal antibody fragments that bind to target cells, or chimeric antibodies, chimeric antibody fragments that bind to target cells, domain antibodies, domain antibody fragments that bind to target cells, adnectins of antibody mimics, DARPins, lymphocyte factors, hormones, vitamins, growth factors, colony stimulating factors, nutrient transport molecules (transferrin), cell-binding peptides, proteins, or small molecules attached to albumin, polymers, dendritic polymers, liposomes, nanoparticles, vesicles, or (viral) capsids.

[0098]

[0099] In another specific embodiment, the cell-binding molecule may be a ligand or receptor agonist selected from the group consisting of: folic acid derivatives (proteins that bind to folic acid receptors and are overexpressed in ovarian cancer and other malignancies) (Low, PS et al. 2008, Acc. Chem. Res. 41, 120-9); glutamate derivatives (proteins that bind to prostate-specific membrane antigens and are surface markers for prostate cancer cells) (Hillier, SM et al. 2009, Cancer Res. 69, 6932-40); somatostatin (also known as growth hormone-inhibiting hormone (GHIH) or growth hormone-releasing inhibitory factor (SRIF) or growth hormone-releasing inhibitory hormone) and analogues such as octreotide (Sandostatin) and somatuline (particularly for neuroendocrine tumors, GH-producing pituitary adenomas, paragangliomas, nonfunctional pituitary adenomas, pheochromocytomas) (Ginj, M. et al. 2006, Proc. Natl. Acad. Sci. USA). 103,16436-41), the following tumors contain somatostatin receptor subtypes (SST1, SST2, SST3, SST4, and SST5): GH-secreting pituitary adenomas (Reubi JC, L and olt, AM 1984 J. Clin. Endocrinol Metab 59: 1148–51; Reubi JC, L and olt AM 1987 J Clin Endocrinol Metab 65: 65–73; Moyse E, et al., J Clin Endocrinol Metab 61: 98–103), gastrointestinal and pancreatic tumors (Reubi JC, et al., 1987 J Clin Endocrinol Metab 65: 1127–34; Reubi, J. C, et al., 1990 Cancer Res 50: 5969–77), pheochromocytomas (Epelbaum J, et al., 1995). J Clin Endocrinol Metab 80:1837–44; Reubi JC et al., 1992 J Clin Endocrinol Metab 74: 1082–9), neuroblastoma (Prevost G, 1996 Neuroendocrinology 63:188–197; Moertel, C. L, et al., 1994 Am J Clin Path 102:752–756), medullary thyroid carcinoma (Reubi, J.C, et al., 1991 Lab Invest 64:567–573), small cell lung cancer (Sagman U, et al., 1990 Cancer 66:2129–2133), meningioma, medulloblastoma or glioma (Reubi JC, et al., 1986 J Clin Endocrinol Metab 63: 433–8; Reubi JC, et al., 1987 Cancer Res 47: 5758–64; Fruhwald, M. C, et al., 1999 Pediatr Res 45: 697–708), breast cancer (Reubi JC, et al., 1990 Int J Cancer 46: 416–20; Srkalovic G, et al., 1990 J Clin Endocrinol Metab 70: 661–669), lymphoma (Reubi JC, et al., 1992, Int J Cancer 50: 895–900), renal cell carcinoma (Reubi JC, et al. 1992, Cancer Res 52: 6074–6078), mesenchymal tumor (Reubi JC, et al. 1996 Cancer Res 56: 1922–31), prostate cancer (Reubi JC, et al. 1995, J. Clin. Endocrinol Metab 80: 2806–14; et al. 1989, Prostate 14:191–208; Halmos G, et al. J. Clin. Endocrinol Metab 85: 2564–71), ovarian cancer (Halmos, G, et al. 2000 J Clin Endocrinol Metab 85: 3509–12; Reubi JC, et al. 1991 Am J Pathol 138:1267–72), gastric cancer (Reubi JC, et al. 1991, Am J Pathol 138:1267–72), gastric cancer (Reubi JC, et al. 1992, Cancer Res 52: 6074–6078), mesenchymal tumor (Reubi JC, et al. 1996, Cancer Res 56: 1922–31), prostate cancer (Reubi JC, et al. 1995, J. Clin. Endocrinol Metab 80: 2806–14; Halmos G, et al. 1989, Prostate 14:191–208; Halmos G, et al. J. Clin. Endocrinol Metab 85: 2564–71), ovarian cancer (Halmos G, et al. 2000, J. Clin. Endocrinol Metab 85: 3509–1 JC, et al. 1999, Int J Cancer 81: 376–86; Miller, G. V, 1992 Br J Cancer 66: 391–95), hepatocellular carcinoma (Kouroumalis E, et al. 1998 Gut 42: 442–7; Reubi JC, et al. 1999 Gut 45: 66–774) and nasopharyngeal carcinoma (Loh K. S, et al. 2002 Virchows Arch 441: 444–8); aromatic sulfonamides (carbonic anhydrase IX specific) (markers of hypoxia and renal cell carcinoma) (Neri, D., et al., Nat. Rev. Drug Discov).2011,10,767-7); Pituitary adenylate cyclase initiating peptide (PACAP) (PAC1) (for pheochromocytoma and paraganglioma); Angioactive intestinal peptide (VIP) and its receptor subtypes (VPAC1, VPAC2); α-melanocyte-stimulating hormone (α-MSH) receptor; Cholecystokinin (CCK) or gastrin receptor and its receptor subtypes (CCK1 (formerly known as CCK-A) and CCK2); Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2) or gastrin-releasing peptide (GRP) and its receptor subclasses (BB1, GRP receptor subclasses (BB2), BB3 and BB4) (Ohlsson, B., et al., 1999, Scand. J. Gastroenterology 34(12): 1224–9; Weber, HC, 2009, Cur. Opin. Endocri. Diab. Obesity 16(1): 66–71, Gonzalez N, et al., 2008, Cur. Opin. Endocri. Diab. Obesity 15(1), 58–64); neurotensin receptors and their receptor subtypes (NTR1, NTR2, NTR3); substance P receptors and their receptor subtypes (e.g., NK1 receptor for gliomas, Hennig IM, et al., 1995, Int. J. Cancer 61, 786–792); neuropeptide Y (NPY) receptors and their receptor subtypes (Y1–Y6); homing peptides include RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimer and polycyclic RGD peptides (e.g., cRGDfV) (Laakkonen P, Vuorinen K. 2010, Integr Biol (Camb). 2(7–8): 326–337; Chen K, Chen X. 2011, Theranostics. 1:189–200; Garanger E, et al., Anti-Cancer Agents Med Chem. 7 (5): 552–558; Kerr, JS et al., Anticancer Research, 19(2A), 959-968; Thumshirn, G, et al., 2003 Chem. Eur. J. 9, 2717-2725), and TAASGVRSMH or LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor) and F3 peptide (31 amino acid peptides that bind to nucleolar receptors expressed on the cell surface) (Zitzmann, S.,2002 Cancer Res.,62,18,5139–5143; Temminga,K.,2005,Drug Resistance Updates, 8,381–402; P. Laakkonen and K. Vuorinen,2010 Integrative Biol,2(7-8),326–337; MA Burg,1999 Cancer Res.,59(12),2869–2874; K. Porkka et al.,2002,Proc. Nat. Acad. Sci. USA 99(11),7444-9); Cell-penetrating peptides (CPPs) (Nakase I, et al.,2012,J. Control Release.159(2), 181–188); peptide hormones, such as agonists and antagonists of luteinizing hormone-releasing hormone (LHRH), gonadotropin-releasing hormone (GnRH) agonists, which act by targeting follicle-stimulating hormone (FSH) and luteinizing hormone (LH) as well as testosterone, such as buserreline (Pyr-His-Trp-Ser-Tyr-D-Ser (OtBu)-Leu-Arg-Pro-NHEt), gosoreline (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2), and gosoreline (Pyr-His-Trp-Ser-Tyr-D-Ser (OtBu)-Leu-Arg-Pro-AzGly-NH2). 2) Histamine resorcinol (Pyr-His-Trp-Ser-Tyr-D-His(N-Bn)-Leu-Arg-Pro-NHEt), Leuprorelin (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), Nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH 2), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH 2), Nafarelin, Dilorine, Aberelin (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropylLys-Pro-DAla-NH) 2), Cetrorelix (Ac-D-2Nal-D-4-chloro-Phe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH 2), Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carba-moyl)-Leu-isopropylLys-Pro-D-Ala-NH 2) And Gannel (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9,N10-diethyl)-homoArg-Leu-(N9,N10-diethyl)-homoArg-Pro-D-Ala-NH 2) (Thundimadathil,J. ,J. Amino Acids,2012,967347,doi:10.1155 / 2012 / 967347; Boccon-Gibod, L.; et al., 2011, Therapeutic Advances in Urology 3(3): 127–140; Debruyne, F., 2006, Future Oncology, 2(6), 677–696; Schally A. V; Nagy, A. 1999 Eur J Endocrinol 141:1–14; Koppan M, et al. 1999 Prostate 38:151–158); Pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), C-type lectins, and nodular receptors (NLRs) (Fukata, M. et al., 2009, Semin. Immunol. 21, 242–253; Maisonneuve, C. et al., 2014, Proc. Natl. Acad. Sci. USA 111, 1–6; Botos, I. et al., 2011, Structure 19, 447–459; Means, TK et al., 2000, Life Sci. 68, 241–258), whose molecular weight ranges from small molecules (imiquimod, guanine, and adenosine analogs) to large and complex biomolecules such as lipopolysaccharide (LPS), nucleic acid (CpG DNA, polyI:C), and lipopeptide (Pam3CSK4) (Kasturi, SP et al., 2011, Nature 470, 543–547; Lane, T., 2001, JR Soc. Med. 94, 316; Hotz, C., and Bourquin, C., 2012, Oncoimmunology 1, 227–228; Dudek, AZ et al., 2007, Clin. Cancer Res. 13,7119–25); Calcitonin receptor, a 32-amino acid neuropeptide, primarily regulates calcium levels through its action on osteoclasts and the kidneys (Zaidi M, et al., 1990 Crit Rev Clin Lab Sci 28, 109–174; Gorn, AH, et al., 1995 J Clin Invest 95: 2680–91); Integrin receptors and their subclasses (such as αVβ1, αVβ3, αVβ5, αVβ6, α6β4, α7β1, αLβ2, αIIbβ3, etc.) typically play important roles in angiogenesis and are expressed on the surface of various cells, particularly osteoclasts, endothelial cells, and tumor cells (Ruoslahti, E.).[References: Albelda, SM et al., 1994 Cell 77, 477-8; Albelda, SM et al., 1990 Cancer Res., 50, 6757-64]. Short peptides, GRGDSPK and cyclic RGD pentapeptides, such as cyclic (RGDfV) (L1) and its derivatives (cyclic (-N(Me)R-GDfV), cyclic (R-Sar-DfV), cyclic (RG-N(Me)D-fV), cyclic (RGD-N(Me)fV), cyclic (RGDf-N(Me)V-)(Silengitide)), have high affinity for integrin receptors (Dechantsreiter, MA et al., 1999 J. Med. Chem. 42, 3033-40; Goodman, SL et al., 2002 J. Med. Chem. 45, 1045-51).

[0100] Cell-binding molecules or ligands or cell receptor agonists can be Ig-based or non-Ig-based protein molecules. Ig-based framework structures can be selected from, but are not limited to, the following groups of components: nanobodies (derives of VHH (cameloid Ig)) (Muyldermans S., 2013 Annu Rev Biochem. 82, 775–97); domain antibodies (dAb, derivatives of VH or VL domains) (Holt, L. J, et al., 2003, Trends Biotechnol. 21, 484–90); bispecific T-cell adapters (BiTE, bispecific biantibody) (Baeuerle, P. A, et al., 2009, Curr. Opin. Mol. Ther. 11, 22–30); dual affinity repositioning agents (DART, bispecific biantibody) (Moore PA P et al., 2011, Blood 117(17), 4542–51); tetravalent tandem antibodies (TandAb, dimeric bispecific biantibody) (Cochlovius, B et al.). 2000, Cancer Res. 60(16):4336–4341).Non-Ig framework structures can be selected from the following groups of components, but are not limited to: Antiticalin (a derivative of Lipocalins) (Skerra A. 2008, FEBS J., 275(11): 2677–83; Beste G et al., 1999 Proc. Nat. Acad. USA. 96(5):1898–903; Skerra, A. 2000 Biochim Biophys Acta, 1482(1-2): 337–50; Skerra, A. 2007, Curr Opin Biotechnol. 18(4): 295–304; Skerra, A. 2008, FEBS J. 275(11):2677–83); Adnectin (the 10th FN3 (fibronectin)) (Koide, A et al., 1998 J. Mol. Biol., 284(4):1141). –51; Batori V, 2002, Protein Eng. 15(12): 1015–20; Tolcher, A. W, 2011, Clin. Cancer Res. 17(2): 363–71; Hackel, B. J, 2010, Protein Eng. Des. Sel. 23(4): 211–19); Designed ankyrin repeat proteins (DARPins) (derived from ankyrin repeat (AR) proteins) (Boersma, YL et al., 2011 Curr Opin Biotechnol. 22(6): 849–57), such as DARPin C9, DARPin Ec4 and DARPin E69_LZ3_E01 (Winkler J et al., 2009 Mol Cancer Ther. 8(9), 2674–83; Patricia MK. M. et al., Clin Cancer Res. 2011; 17(1):100–10; Boersma Y. L et al., 2011 J. Biol. Chem. 286(48),41273–85); Avimer (domain A / low-density lipoprotein (LDL) receptor)(Boersma Y. L., 2011 J. Biol. Chem. 286(48): 41273–41285; Silverman J et al., 2005 Nat. Biotechnol.,23(12):1556–61).

[0101] Examples of small molecule structures of cell-binding molecules / ligands or cell receptor agonists in patent applications are as follows: LB01 (folic acid), LB02 (PMSA ligand), LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (somatostatin), LB06 (somatostatin), LB07 (octreotide, somatostatin analog), LB08 (lanreotide, somatostatin analog), LB09 (Sanvar, somatostatin analog), LB10 (CAIX ligand), LB11 (CAIX ligand), LB12 (gastrin-releasing peptide receptor (GRPr), MBA), LB13 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligand), LB14 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligand), LB15 ( GnRH antagonists (Abarelix), LB16 (cobalamin, vitamin B12 analog), LB17 (cobalamin, vitamin B12 analog), LB18 (for αvβ3 integrin receptor, cyclic RGD pentapeptide), LB19 (VEGF receptor isodivalent peptide ligand), LB20 (neuromycin B), LB21 (froglin, acting on G protein-coupled receptors), LB22 (TLR2, acting on Toll-like receptors), LB23 (acting on androgen receptors), LB24 (silengiptide or cyclic (-RGDfV-) conjugate, acting on αvβ3 integrin receptors). (v integrin receptor), LB23 (flucodone), LB25 (rifabutin analog), LB26 (rifabutin analog), LB27 (rifabutin analog), LB28 (fludrocortisone), LB29 (dexamethasone), LB30 (fluticasone propionate), LB31 (beclomethasone propionate), LB32 (triamcinolone acetate), LB33 (prednisolone), LB34 (prednisolone), LB35 (methylprednisolone), LB36 (betamethasone), LB37 (irinotecan analog), LB38 (crizotinib analog), LB39 The following analogues are listed: LB40 (bortezomib analogue), LB41 (carfilzomib analogue), LB42 (leuprorelin analogue), LB43 (triptorelin analogue), LB44 (clindamycin), LB45 (liraglutide analogue), LB46 (vincristine analogue), LB47 (retapalline analogue), LB48 (butulin analogue), LB49 (vincristine analogue), LB50 (lixisentide analogue), LB51 (osidinib analogue), LB52 (nucleoside analogue), LB53 (erlotinib analogue), and LB54 (lapatinib analogue). Their structures are shown below: LB01 (Folic Acid), LB02 (PMSA ligand), LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (Somatostatin), LB06 (Somatostatin), LB07 (octreotide, a somatostatin analogue) LB08 (lanopeptide, somatostatin analogue), LB09 (amino peptide (Sanvar), a somatostatin analogue), LB10 (CAIX ligand), LB11 (CAIX ligand), LB12 (gastrin-releasing peptide receptor (GRPr), MBA) LB13 (LH-RH and GnRH ligands), LB14 (LH-RH and Gonadotropin-releasing hormone GnRH ligand), LB15 (GnRH antagonist, abaric), R 19 is 5'-deoxyadenosine, Me, OH, CN; LB16 (cobalamin, a vitamin B12 analog). R 19 is 5'-deoxyadenosine, Me, OH, CN; LB17 (cobalamin, a vitamin B12 analog). LB18 (cyclic RGD pentapeptide, acting on αvβ3 integrin receptor), LB19 (a heterodivalent peptide ligand conjugate that acts on the vascular endothelial growth factor (VEGF) receptor) LB20 (neuromucoid B), LB21 (frog dermalin conjugate, acting on G protein-coupled receptors) LB22 (TLR2 conjugate, acting on Toll receptor-like receptors), LB23 (androgen receptor), LB24 (Cyrenogen / cyclic (-RGDfV-) conjugate, acting on αv integrin receptor) LB25 (Rifabutin analogue), LB26 (Rifabutin analogue), LB27 (Rifabutin analogue), LB28 (Fluordrocortisone), LB29 (Dexamethasone) LB30 (fluticasone propionate), LB31 (beclomethasone propionate), LB32 (triamcinolone acetonide), LB33 (Prednisone) LB34 (Prednisolone) LB35 (methylprednisolone) LB36 (Beflumethasone), LB37 (irinotecan analogue), LB38 (crizotinib analogue), LB39 (bortezomib analogue), wherein Y5 is N, CH, C(Cl), C(CH3) or C(COOR1); R1 is H, C1-C6 alkyl, C3-C8 aryl; LB40 (cafizzomi analogue), LB41 (cafizzomi analogue), LB42 (leuprolide analogue), LB43 (triptorelin analogue), LB44 (Clindamycin), LB45 (liraglutide analogue), LB46 (Semaglutide analogue), LB47 (Ritaparin analogue), LB48 (Dimbulin analogue), LB49 (vincrine analogue), LB50 (Lixisen peptide analog), LB51 (Ositinide analogue), LB52 (nucleoside analogue), LB53 (erlotinib analogue), LB54 (lapatinib analogue), in" “ represents the connection position of the linker; X4 and Y1 are independently O, NH, NHNH, NR1, S, C(O)O, C(O)NH, OC(O)NH, OC(O)O, NHC(O)NH, NHC(O)S, OC(O)N(R1), N(R1)C(O)N(R1), CH2, C(O)NHNHC(O) and C(O)NR1; X1 is H, CH2, OH, O, C(O), C(O)NH, C(O)N(R1), R1, NHR1, NR1, C(O)R1 or C(O)O; X5 is H, CH3, F or Cl; M1 and M2 are independently H, Na, K, Ca, Mg, NH4, N(R1R1'R2R3); R 1. R1', R2 and R3 are defined in the same way as equation (I).

[0102] Preparation of couplings

[0103] In another embodiment of the invention, preferably, the synthesized camptothecin analogue contains a linker L and a reactive group Lv, as shown in formulas (VI), (VII), (VIII), (IX), and (X), and can readily react with cell-binding molecule T or a modified cell-binding molecule T to form conjugates as shown in formulas (I), (II), (III), (IV), and (V), respectively: (VI) (VII) (VIII) (IX) (X) Where R1, R2, R3, R4, R5, L, X and m are the same as those defined in equation (I) above; Lv is a reactive functional group that can react with thiols, amines, carboxylic acids, selenools, phenols, or hydroxyl groups on cellular molecules. It includes, but is not limited to, halogens (such as fluorine, chlorine, bromine, iodine); maleimino; methanesulfonyl; toluenesulfonyl; trifluoromethanesulfonyl; nitrophenoxy; N-hydroxysuccinimino (NHS); phenoxy; dinitrophenoxy; pentafluorophenoxy; tetrafluorophenoxy; trifluorophenoxy; difluorophenoxy; monofluorophenoxy; pentachlorophenoxy; imidazole; monochlorophenoxy; dichlorophenoxy; trichlorophenoxy; tetrachlorophenoxy Phenoxy; N-(benzotriazolyl)oxy; 2-ethyl-5-phenylisooxazolyl-3′-sulfonyl; phenoxydiazolylsulfonyl; 2-ethyl-5-phenylisooxazolyl; phenoxydiazolyl; oxadiazolyl; unsaturated carbon (double or triple bonds between carbon-carbon, carbon-nitrogen, carbon-sulfur, carbon-phosphorus, sulfur-nitrogen, phosphorus-nitrogen, oxygen-nitrogen, or carbon-oxygen); or intermediates formed by reaction with Mitsunobu reagent. Examples of condensing agents are as follows: 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC), dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), N-cyclohexyl-N'-(2-morpholino-ethyl)carbodiimide methyl p-toluenesulfonate (CMC or CME-CDI), 1,1'-carbonyldiimidazole (CDI), (O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU), N,N,N',N'-tetramethyl-oxo-(1H-benzotriazol-1-yl)-ammonium hexafluorophosphate (HBTU), ( Benztriazol-1-yloxy)tris(dimethylamino)-hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinylhexafluorophosphate (PyBOP), diethyl cyanophosphonate (DEPC), chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine-1-oxohexafluorophosphate (HATU), 1-[(dimethylamino)(morpholino)methylene]-1H-[1,2,3]triazolo[4,5-b]pyridine-1-onthium-3-oxohexafluorophosphate (HDMA), 2-chloro-1,3-dimethyl-imidazolium hexafluorophosphate (CIP), chloropyrrolidone-onthium hexafluorophosphate (PyCloP), fluorine- N,N,N',N'-bis(tetramethylene)formamidinium hexafluorophosphate (BTFFH), N,N,N',N'-tetramethyl-S-(1-sideoxy-2-pyridyl)thiourea hexafluorophosphate, O-(2-sideoxy-1(2H)pyridyl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TPTU), S-(1-sideoxy-2-pyridyl)N,N,N',N'-tetramethylthiourea tetrafluoroborate, O-[(ethoxycarbonyl)-cyanomethylamino]-N,N,N',N'-Tetramethylurea hexafluorophosphate (HOTU), (1-cyano-2-ethoxy-2-sideoxyethylaminooxy)dimethylamino-morpholino-hexafluorophosphate (COMU), O-(benzotriazol-1-yl)-N,N,N',N'-bis(tetramethylene)hexafluorophosphate (HBPyU), N-benzyl-N'-cyclohexyl-carbodiimide (with or without polymer binding), dipyrrolidyl(N-succiniminooxy)carbomonium hexafluorophosphate (HSPyU), dichloropyrrolidyl hexafluorophosphate (PyClU), 2-chloro-1,3-dimethylimidazolium tetrafluoroborate (CIB), (benzotriazol-1-yloxy)diguanidine carbomonium hexafluorophosphate (HBPipU), O-(6-chlorobenzotriazol-1-yl)- N,N,N',N'-Tetramethylurea tetrafluoroborate (TCTU), bromo(dimethylamino)-hexafluorophosphate (BroP), propylphosphonic anhydride (PPACA, T3P), 2-morpholinoethyl isocyanate (MEI), N,N,N',N'-tetramethyl-oxo-(N-succinimino)hexafluorophosphate (HSTU), 2-bromo-1-ethyl-pyridinium tetrafluoroborate (BEP), O-[(ethoxycarbonyl)cyano-methyleneamino]-N,N,N',N'-tetramethylurea tetrafluoroborate (TOTU), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine chloride (MMTM, DMTMM), N,N,N',N'-tetramethyl-oxo-(N-succinimino) Urea tetrafluoroborate (TSTU), O-(3,4-dihydro-4-sideoxy-1,2,3-benzotriazine-3-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TDBTU), 1,1'-(azodicarbonyl)-diguanidine (ADD), di-(4-chlorobenzyl)azodicarboxylate (DCAD), ditert-butyl azodicarboxylate (DBAD), diisopropyl azodicarboxylate (DIAD), and diethyl azodicarboxylate (DEAD). Additionally, Lv can be an anhydride, an anhydride formed by the self-condensation of an acid or by the reaction of an acid with other C1-C8 anhydrides; Preferred Lv compounds include the following: halides (such as fluorides, chlorides, bromides, and iodides), mesyl, tosyl, trifluoromethylsulfonyl, trifluoromethylsulfonate, nitrophenoxy, N-succinimino (NHS), phenoxy; dinitrophenoxy; pentafluorophenoxy, tetrafluorophenoxy, trifluorophenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, 1H-imidazol-1-yl, monochlorophenoxy, dichlorophenoxy, trichlorophenoxy, tetrachlorophenoxy, N-(benzotriazolyl)oxy, 2-ethyl-5-phenylisoxazole-3′- Sulfoyl, phenyloxadiazole-sulfoyl, 2-ethyl-5-phenylisooxazolyl, phenyloxadiazole, oxadiazole, unsaturated carbon (double or triple bonds between carbon-carbon, carbon-nitrogen, carbon-sulfur, carbon-phosphorus, sulfur-nitrogen, phosphorus-nitrogen, oxygen-nitrogen, or carbon-oxygen), or one of the following structures: Disulfides; Halogenated acetylated; Brine; N-hydroxysuccinimide ester; Maleimide; Monosubstituted maleic anhydride; Disubstituted maleic anhydride; Monosubstituted succinimides; Disubstituted maleic anhydride; Substitute maleic acid; -CHO aldehyde; Ethylenesulfonyl; Acrylic; 2-(Toluenesulfonoxy)acetyl; 2-(methanesulfonyloxy)acetyl; 2-(nitrophenoxy)acetyl; 2-(dinitrophenyl)acetyl; 2-(Fluorophenoxy)-acetyl; 2-(difluorophenoxy)-acetyl; 2-((trifluoromethylsulfonyl)oxy)acetyl; Ketones or aldehydes 2-(pentafluorophenoxy)acetyl; Methylphenyl oxadiazole (ODA); , acid anhydride, Alkoxyamine; Azide group, Alkyne group, or Azoxyhydrazine; wherein X1' is F, Cl, Br, I, or Lv3; X2' is O, NH, N(R1), or CH2; R3 is H, aryl, or heteroaryl, wherein one or more H atoms may be independently substituted by -R1, -halogen, -OR1, -SR1, -NR1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1; Lv3 is a leaving group selected from the group consisting of F, Cl, Br, I; nitrophenoxy; N-hydroxysuccinylimine (NHS); phenoxy; dinitrophenoxy; pentafluorophenoxy; tetrafluorophenoxy; difluorophenoxy; monofluorophenoxy; pentachlorophenoxy; trifluoromethanesulfonyl; imidazolyl; dichlorophenoxy; tetrachlorophenoxy; 1-Hydroxybenzotriazolyl; toluenemethanesulfonyl; methanesulfonyl; 2-ethyl-5-phenylisoxazole-3′-sulfonyl, anhydride or anhydride formed by reaction with other anhydrides, such as acetic anhydride, formic anhydride; or intermediate formed by reaction with polypeptide condensation reagents, Mitsunobu reagent.

[0104] During the coupling process, prior to coupling with camptothecin analogs, cell-binding molecules can be modified by using linkers containing bifunctional groups to link more specific peptides, proteins, drug molecules, or other functional molecules. These linkers containing bifunctional groups include linkers linking amines to non-selective functional groups (succinimide ester (NHS ester)-diazacyclopropene (SDA), succinimide ester-azide), linkers linking amines to thiol groups (NHS ester-maleimide, NHS ester-pyridinedithio, NHS ester-haloacetyl), linkers linking thiols to sugars (maleimide-acetylhydrazine, pyridinedithio-acetylhydrazine), linkers linking hydroxyl to thiol groups (isocyanate-maleimide), linkers linking amines to DNA (NHS ester / psoralen), and linkers linking amines to carboxyl groups (carbodiimide), etc.

[0105] When using the succinimide (NHS)-diazacyclopropene (SDA) linker, the NHS ester on the linker first reacts with the amine on the cellular binding molecule backbone (in a buffer solution at pH 6-9) to form a stable amide bond. Then, under long-wavelength irradiation at 330-370 nm, the diazacyclopropene is activated to produce a carbene reactive intermediate, which then reacts with the amine on a specific peptide, protein, or other functional molecule. The order of these two reactions can also be altered: first, the amine on the functional molecule reacts with the NHS ester on the linker, and then chemically reacts with the cellular binding molecule under light (330-370 nm) irradiation. The succinimide (NHS)-diazacyclopropene (SDA) linker can also be cleavable (like the SDAD linker, containing internal disulfide bonds).

[0106] When using NHS ester / azide linkers, the NHS ester can first react with the amine on the cellular binding molecular backbone in a buffer solution at pH 6-9 to form a stable amide bond. Then, via the Huisgen azido-alkyne cycloaddition reaction, the alkynyl group on a specific polypeptide, protein, or other functional molecule reacts with the azide group at the other end of the crosslinker to form a 1,2,3-triazole (click chemistry). Alternatively, the NHS ester can also first react with the amine on the functional molecule in a buffer solution at pH 6-9 to form a stable amide bond, and then the alkynyl group on the binding molecule undergoes a Huisgen azido-alkyne cycloaddition reaction with the azide group at the other end of the linker to form a 1,2,3-triazole linker.

[0107] In amine-thiol linkage, NHS esters can first react with the amine on the cellular binding molecule backbone in a buffer solution at pH 6–9 to form a stable amide-amine bond. Then, at pH 4.5–8.5, the thiol groups on more specific peptides, proteins, or other functional molecules react with the maleimine, pyridine dimercapto, or haloacetyl group at the other end of the linker to form a thioether or disulfide bond. The order of the coupling reaction can also be altered. For example, the amine group on the functional molecule can first react with the linker to form an amide-amine bond, and then react with the thiol group on the cellular binding molecule. Alternatively, at pH 4.5–7.0, the thiol group on the functional molecule can first react with the linker to form a thioether or disulfide bond, and then react with the amine group on the cellular binding molecule at pH 6–9 to form an amide-amine bond.

[0108] In thiol-glycan linkages, under pH conditions of 4.5–8, the thiol group on the cell-binding molecule can first react with maleimide or pyridine dimercapto to form a thioether or disulfide bond. Then, the carbonyl group (aldehyde / ketone) on the functional molecule reacts with acehydrazine to form a hydrazone bond. Alternatively, under pH conditions of 4.5–8, the thiol group on the functional molecule can also first react with the linker to form a thioether or disulfide bond, and then react with the sugar, oxidized sugar, or carbonyl group (aldehyde / ketone) on the cell-binding molecule to form a hydrazone bond.

[0109] In the case of hydroxyl-thiol linkage, under conditions of pH 6–8, the thiol group on the cell-binding molecule can first react with maleimide or pyridine dimercapto to form a thioether or disulfide bond. Then, under conditions of pH 8–9, the hydroxyl group on the functional molecule reacts with isocyanate to form a carbamate. Alternatively, the thiol group on the functional molecule can also first react with the linker under conditions of pH 6–8 to form a thioether or disulfide bond. Then, it reacts with the hydroxyl group on the cell-binding molecule under conditions of pH 8–9 to form a carbamate.

[0110] In another embodiment of the invention, the production of antibodies used in the invention comprises in vivo or in vitro methods or combinations thereof. Methods for producing polyclonal anti-receptor peptide antibodies are well known in the art, for example, as described in U.S. Patent 4,493,795. Monoclonal antibodies are typically prepared by fusing myeloma cells with spleen cells from mice already immunized with the desired antigen (Köhler, G.; Milstein, C. Nature 1975, 256: 495-497). Detailed procedures are described in “Antibodies—A Laboratory Manual, Harlow and Lane, eds., Cold Spring Harbor Laboratory Press, New York (1988),” which is incorporated herein by reference. Specifically, antibodies can be prepared by immunizing mice, rats, hamsters, or any other mammals with target antigens, such as antigens isolated from target cells, intact viruses, inactivated whole viruses, and viral proteins. Polyethylene glycol (PEG) 6000 is typically used to fuse spleen cells with myeloma cells. Hybrid cells are screened by their sensitivity to HAT (hypoxanthine-aminopterin-thymine). Hybridomas carrying out the monoclonal antibodies of this invention can be identified by their ability to receive immune-specific receptors or to inhibit receptor activity on target cells.

[0111] The production of the monoclonal antibodies used in this invention is carried out in a monoclonal hybridoma culture, which contains a nutrient medium and a hybridoma capable of secreting antibody molecules with suitable antigen specificity. The culture is maintained under suitable conditions for a sufficient period of time to allow the hybridoma to secrete antibody molecules into the medium. The medium containing the antibody is then collected. The antibody molecules are further separated using known techniques, such as protein A affinity chromatography, anionic, cationic, hydrophobic, or size exclusion chromatography (particularly by protein A affinity chromatography and size exclusion chromatography), centrifugation, differential solubility, or any other standard technique for purifying proteins.

[0112] Culture media used to prepare these compositions are well known in the industry and are commercially available, including synthetic media. An example of a synthetic media is Dulbecco Minimal Essential Medium (DMEM; Dulbecco et al., Virol. 1959, 8, 396), supplemented with 4.5 g / ml glucose, 20 mM glutamine, 20% fetal bovine serum, and antifoaming agents such as polyoxyethylene-polyoxypropylene block copolymers.

[0113] In addition, antibody-producing cell lines can also be obtained through techniques other than fusion, such as transplanting tumorigenic DNA into B lymphocytes, or transfecting with tumorigenic viruses, such as Epstein-Barr virus (EBV, also known as human herpesvirus 4 (HHV-4)) or Kaposi's sarcoma-associated herpesvirus (KSHV), as described in U.S. Patents 4,341,761; 4,399,121; 4,427,783; 4,444,887; 4,451,570; 4,466,917; 4,472,500; 4,491,632; and 4,493,890. Monoclonal antibodies can also be prepared from antireceptor peptides or peptides containing terminal carboxyl groups, as is well known in the field. See Niman et al., Proc. Natl. Acad. Sci. USA, 1983, 80: 4949-4953; Geysen et al., Proc. Natl. Acad. Sci. USA, 1985, 82: 178-182; Lei et al. Biochemistry 1995, 34(20): 6675-6688. Typically, as immunogens for generating antireceptor peptide monoclonal antibodies, antireceptor peptides or peptide analogs can be used alone or linked to an immunogenic carrier.

[0114] The monoclonal antibodies used as binding molecules in this invention can also be obtained using other techniques known in the industry. A particularly useful method is the production of fully human antibodies. One method is phage display, which uses affinity enhancement to select human antibodies that specifically bind to the antigen. Phage display technology is described in detail in the literature, and the construction and screening of phage display libraries are well-known in the industry. See Dente et al., Gene. 1994, 148(1):7-13; Little et al., Biotechnol Adv. 1994, 12(3):539-55; Clackson et al., Nature 1991, 352:264-628; Huse et al., Science 1989, 246:1275-1281.

[0115] Monoclonal antibodies produced by hybridomas fused with non-human cells, such as mouse cells, can be humanized to avoid the production of human anti-mouse antibodies. Common antibody humanization methods include complementarity-determining region (CDR) transplantation, which have been described in detail in U.S. Patents 5,859,205 and 6,797,492; Liu et al., Immunol Rev. 2008, 222:9-27; Almagro et al., Front Biosci. 1; 2008, 13: 1619-33; Lazar et al., Mol Immunol. 2007, 44(8):1986-98; Li et al., Proc. Natl. Acad. Sci. US A. 2006, 103(10):3557-62, which are incorporated herein by reference. Complete human antibodies can also be prepared by immunizing transgenic mice, rabbits, monkeys, or other mammals carrying most of the human globulin heavy and light chains with an immunogen. Examples of these mice include Xenomouse (Abgenix, Inc.), HuMAb-Mouse (Medarex / BMS), and VelociMouse (Regeneron), see U.S. Patents 6,596,541,6,207,418,6,150,584,6,111,166,6,075,181,5,922,545,5,661,016,5,545,806,5,436,149, and 5,569,825. For human treatment, the variable region from mice and the constant region from humans can also be fused to form a "chimeric antibody," which exhibits significantly lower immunogenicity in humans than mouse monoclonal antibodies (Kipriyanov et al., Mol Biotechnol. 2004, 26:39-60; Houdebine, Curr Opin Biotechnol. 2002, 13:625-9). Furthermore, site-directed mutations in the variable regions of antibodies can lead to higher affinity and specificity (Brannigan et al., Nat Rev Mol Cell Biol. 2002, 3:964-70; Adams et al., J Immunol Methods. 1999, 231:249-60), and alterations in the constant regions of mAbs can enhance their effector functions in mediating binding and cytotoxicity.

[0116] Immunospecific antibodies against malignant tumor cell antigens can be obtained commercially or produced by any known method, such as chemical synthesis or recombinant expression techniques. The nucleotide sequences encoding antibodies with immunospecificity against malignant tumor cell antigens are commercially available, for example, from GenBank or similar databases, literature publications, or from known cloning and sequencing.

[0117] Besides antibodies, a peptide or protein that interacts with antigenic determinants or corresponding receptors on target cells (binding, blocking, targeting, or other types of interaction) can also serve as a binding molecule. These peptides or proteins can be any random peptide or protein that has an affinity for antigenic determinants or corresponding receptors and does not necessarily have to be members of the immunoglobulin family. These peptides can be isolated using techniques similar to phage display of antibodies (Szardenings, J Recept Signal Transduct Res. 2003; 23(4):307-49). Similar to antibodies and antibody fragments, peptides obtained from random peptide libraries can be coupled or linked to macromolecules or other substances, including but not limited to albumins, polymers, liposomes, nanoparticles, and dendritic molecules. Such linkages should preserve the antigen-binding specificity of the peptide or protein.

[0118] Examples of antibodies in camptothecin analogue conjugates used to treat cancer, autoimmune diseases, and / or infectious diseases include, but are not limited to, 3F8 (anti-GD2), abamumab (anti-CA-125), abciximab (anti-CD41, integrin α-IIb), adalimumab (anti-TNF-α), adecatumumab (anti-EpCAM, CD326), afimomumab (anti-TNF-α), afutuzumab (anti-CD20), alacizumab (anti-VEGFR2), ALD518 (anti-IL-6), alemtuzumab (Campath, MabCampath, anti-CD52), attumomab (anti-CEA), anatumomab (anti-TAG-72), anrukinzumab (IMA-638, anti-IL-13), and apolizumab. (Anti-HLA-DR), Azithromycin (anti-CEA), Asceticumab (anti-L-selectin, CD62L), Atlizumab (tocilizumab, Actemra, RoActemra, anti-IL-6 receptor), Atorolimumab (anti-Rhesus factor), Bapineuzumab (anti-β-amyloid), Basiliximab (Simulect, anti-CD25 (α chain of IL-2 receptor)), Bavituximab (anti-phosphatidylserine), Bectomomab (LymphoScan, anti-CD22), Benlysta (LymphoStat-B, anti-BAFF), Benralizumab (anti-CD125), Bertilimumab (anti-CCL11 (eotaxin-1)), Besilesomab (Scintimun, anti-CEA-associated antigen) Bevacizumab (Avastin, anti-VEGF-A), Biciromab (FibriScint, anti-fibrin II β chain), Bivatuzumab (anti-CD44v6), Blinatumomab (BiTE, anti-CD19), Brentuximab (cAC10, anti-CD30 TNRSF8), Briakinumab (anti-IL-12, IL-23), Canakinumab (Ilaris, anti-IL-1), Cantuzumab (C242, anti-CanAg), Capromab, Catumaxomab (Removab, anti-EpCAM, anti-CD3), CC49 (anti-TAG-72), Cedelizumab (anti-CD4), Certolizumab monoclonal antibody (Cimzia, anti-TNF-α).Cetuximab (Erbitux, IMC-C225, anti-EGFR), Citatuzumab bogatox (anti-EpCAM), Cixutumumab (anti-IGF-1), Clenoliximab (anti-CD4), Clivatuzumab (anti-MUC1), Conatumumab (anti-TRAIL-R2), CR6261 (anti-influenza A hemagglutinin), Dacetuzumab (anti-CD40), Daclizumab (Zenapax, anti-CD25 (IL-2 receptor α chain)), Daratumumab (anti-CD38 (cyclic ADP ribohydrolase)), Denosumab (Prolia, anti-RANKL), Detumomab (anti-B lymphoma) Cellular), Dorlimomab, Dorlixizumab, Ecromeximab (anti-GD3 ganglioside), Eculizumab (Soliris, anti-C5), Edobacomab (anti-endotoxin), Edrecolomab (Panorex, MAb17-1A, anti-EpCAM), Efalizumab (Raptiva, anti-LFA-1 (CD11a)), Efungumab (Mycograb, anti-Hsp90), Elotuzumab (anti-SLAMF7), Elsilimomab (anti-IL-6), Enlimomab Monoclonal antibodies (anti-ICAM-1 (CD54)), Epitumomab (anti-episialin), Etazobumab (anti-CD22), Erlizumab (anti-ITGB2 (CD18)), Ertumaxomab (Rexomun, anti-HER2 / neu, CD3), Etazobumab (Abegrin, anti-integrin αvβ3), Exbivirumab (anti-hepatitis B surface antigen), Fanolesomab (NeutroSpec, anti-CD15), Faralimomab (anti-interferon receptor), Farletuzumab (anti-folate receptor 1), Felvizumab (anti-respiratory syncytial virus), Fezakinumab (Anti-IL-22), Figitumumab (anti-IGF-1 receptor), Fontolizumab (anti-IFN-γ), Foravirumab (anti-rabies virus glycoprotein), Fresolimumab (anti-TGF-β), Galiximab (anti-CD80), Gantenerumab (anti-β-amyloid), Gavilimomab (anti-CD147 (basigin)), Gemtuzumab (anti-CD33), Girentuximab (anti-carbonic anhydrase 9).Glembatumumab (CR011, anti-GPNMB), Golimumab (Simponi, anti-TNF-α), Gomiliximab (anti-CD23 (IgE receptor)), anti-HLA-DR antibody, Ibalizumab (anti-CD4), Ibritumomab (anti-CD20), Igovomab (Indimacis-125, anti-CA-125), Imciromab (Myoscint, anti-myosin), Infliximab (Remicade, anti-TNF-α), Intetumumab (anti-CD51), Inolimomab (anti-CD25 (IL-2 receptor α chain), Izumab (anti-CD22), Ipilimumab (anti-CD152), Iratumumab (anti-CD30 (TNFRSF8)), Keliximab (anti-CD4), Labetuzumab (CEA-Cide, anti-CEA), Lebrikizumab (Anti-IL-13), Lemalesomab (anti-NCA-90 (granulocyte antigen)), Lerdelimumab (anti-TGF) β2), Lexatumumab (anti-TRAIL-R2), Libivirumab (anti-hepatitis B surface antigen), Lintuzumab (anti-CD33), Lumilimumab (anti-CD40), Lumilimumab (anti-CD23 (IgE receptor)), Mapatumumab (anti-TRAIL-R1), Massimumab (anti-T-cell receptor), Matozumab (anti-EGFR), Mepolizumab (Bosatria, anti-IL-5), Metelimumab (anti-TGFβ1), Milatuzumab (anti-CD74), Minretumomab (anti-TAG-72), Mitumomab (BEC-2, anti-GD3 ganglioside), Morolimumab (anti-rhesus monkey factor), Motavizumab (Numax, anti-respiratory syncytial virus), Muromonab-CD3 (Orthoclone OKT3, anti-CD3), Nacolomab (anti-C242), Naptumomab (Anti-5T4), Natazumab (Tysabri, anti-integrin α4), Nabatabumab (anti-endotoxin), Necitumumab (anti-EGFR), Nerelimomab (anti-TNF-α), Nimotuzumab (Theracim, Theraloc, anti-EGFR), Nofetumomab, Ocrelizumab (anti-CD20), Afolimomab (anti-LFA-1 (CD11a)).Ofatumumab (Arzerra, anti-CD20), Olaratumab (anti-PDGF-R α), Omalizumab (Xolair, anti-IgE Fc region), Oportuzumab (anti-EpCAM), Oregovomab (OvaRex, anti-CA-125), Otelixizumab (anti-CD3), Pagibaximab (anti-lipoteichoic acid), Palivizumab (Synagis, Abbosynagis, anti-respiratory syncytial virus), Panitumumab (Vectibix, ABX-EGF, anti-EGFR), Panobacumab (anti-Pseudomonas aeruginosa) ), Percoizumab (anti-IL-4), Pemtumomab (Theragyn, anti-MUC1), Pertuzumab (Omnitarg, 2C4, anti-HER2 / neu), Pexelizumab (anti-C5), Pintumomab (anti-adenocarcinoma antigen), Priliximab (anti-D4), Pritumumab (anti-vimentin), PRO140 (anti-CCR5), Racotumomab (1E10, anti-N-hydroxyacetylneuraminic acid (NeuGc, NGNA)-ganglioside GM3), Rafivirumab (anti-rabies virus glycoprotein), Ramucirumab (anti-VEGFR2), Ranibizumab (Lucentis, anti-VEGF-A), Raxibacumab (Anti-anthrax toxin, protective antigen), Regavirumab (anti-cytomegalovirus glycoprotein B), Reslizumab (anti-IL-5), Rilotumumab (anti-HGF), Rituximab (MabThera, Rituxanmab, anti-CD20), Robatumumab (anti-IGF-1 receptor), Rontalizumab (anti-IFN-α), Rovelizumab (LeukAr-rest, anti-CD11, CD18), Ruplizumab (Antova, anti-CD154 (CD40L)), Satumomab (anti-TAG-72), Sevirumab (anti-cytomegalovirus), Sibrotuzumab (anti-FAP), Siframumab (anti-IFN-α), Siltuximab (anti-IL-6), Siplizumab (anti-CD2), MI 95 (anti-CD33), Solanezumab (anti-β-amyloid), Sonepizumab (sphingosine 1-phosphate), Sontuzumab (anti-episialin),Stamulumab (anti-myostatin), Sulesomab (LeukoScan, anti-NCA-90 (granulocyte antigen)), Tacatuzumab (anti-alpha-fetoprotein), Tadocizumab (anti-integrin αIIbβ3), Talizumab (anti-IgE), Tanezumab (anti-NGF), Taplitumomab (Anti-CD19), Tefibazumab (Aurexis, (anti-agglutination factor A)), Telimomab, Tenatumomab (anti-tenosynovin C), Teneliximab (anti-CD40), Teplizumab (anti-CD3), TGN1412 (anti-CD28), Ticilimumab (Tremelimumab, anti-CTLA-4), Tigatuzumab (anti-TRAIL-R2), TNX-650 (anti-IL-13), Tocilizumab (Atlizumab, Actemra, RoActemra, anti-IL-6 receptor), Toralizumab (anti-CD154 (CD40L)), Tositumomab (anti-CD20), Trastuzumab (Herceptin, anti-HER2 / neu), Tremelimumab (anti-CTLA-4), Tucotuzumab celmoleukin (anti-EpCAM) Tuvirumab (anti-hepatitis B virus), Urtoxazumab (anti-E. coli), Ustekinumab (Stelara, anti-IL-12, IL-23), Vapaliximab (anti-AOC3 (VAP-1)), Vedolizumab (anti-integrin α4β7), Vedolizumab (anti-CD20), Vepalimomab (anti-AOC3 (VAP-1)), Visilizumab (Nuvion, anti-CD3), Vitaxin (anti-angiotensin avb3), Volociximab (anti-integrin α5β1), Votumumab (HumaSPECT, anti-tumor) Antigen CTAA16.88), Zalutumumab (HuMax-EGFR, anti-EGFR), Zanolimumab (HuMax-CD4, anti-CD4), Ziralimumab (anti-CD147 (basigin)), Zolimomab (anti-CD5), Etanercept (Enbrel®), Alefacept (Amevive®), Abatacept (Orencia®), Rilonacept (Arcalyst), 14F7 (anti-IRP-2 (ferrregular protein 2)), 14G2a (anti-GD2 ganglioside,(Derived from Nat. Cancer Institute, for the treatment of melanoma and solid tumors), J591 (anti-PSMA, derived from Weill Cornell Medical College, for the treatment of prostate cancer), 225.28S (anti-HMW-MAA (high molecular weight melanoma-associated antigen)), Sorin Radiofarmaci SRL (derived from Milan, Italy, for the treatment of melanoma), COL-1 (anti-CEACAM3, CGM1, derived from Nat. Cancer Institute, for the treatment of colorectal and gastric cancer), CYT-356 (Oncoltad®, for the treatment of prostate cancer), HNK20 (OraVax Inc., for the treatment of respiratory syncytial virus infection), ImmuRAIT (derived from Immunomedics, for the treatment of NHL), Lym-1 (anti-HLA-DR10, Peregrine) Pharm (for cancer treatment), MAK-195F (anti-TNF (tumor necrosis factor, TNFA, TNF-α, TNFSF2), derived from Abbott / Knoll, for the treatment of septic shock), MEDI-500 (T10B9, anti-CD3, TRαβ (T cell receptor α / β), derived from MedImmune Inc, for the treatment of graft-versus-host disease), RING SCAN (anti-TAG 72 (tumor-associated glycoprotein 72), derived from Neoprobe). Corp., for the treatment of breast, colon, and rectal cancer), Avicidin (anti-EPCAM (epithelial cell adhesion molecule)), anti-TACSTD1 (tumor-associated calcium signaling 1), anti-GA733-2 (gastrointestinal tumor-associated protein 2), anti-EGP-2 (epithelial glycoprotein 2), anti-KSA, KS1 / 4 antigen, M4S, tumor antigen 17-1A, CD326 (from NeoRx, for the treatment of colon, ovarian, prostate, and NHL), anti-Trop2 humanized antibody hRS7, LymphoCide (from Immunomedics), Smart ID10 (from Protein Design Labs), Oncolym (from Techniclone Inc.), Allomune (from BioTransplant), anti-VEGF (from Genentech), CEAcide (from Immunomedics), IMC-1C11 (from ImClone Systems), and Cetuximab (from ImClone).

[0119] Other antibodies that can act as ligands include, but are not limited to, antibodies against the following antigens: aminopeptidase N (CD13), annexin A1, B7-H3 (CD276, various cancers), CA125 (ovarian cancer), CA15-3 (various cancers), CA19-9 (various cancers), L6 (various cancers), Lewis Y (various cancers), Lewis X (various cancers), alpha-fetoprotein (various cancers), CA242 (colorectal cancer), placental alkaline phosphatase (various cancers), prostate-specific antigen (prostate cancer), prostate acid phosphatase (prostate cancer), epidermal growth factor (various cancers), CD2 (Hodgkin's disease, NHL lymphoma, multiple myeloma), CD3ε (T-cell lymphoma, lung cancer, breast cancer, gastric cancer, ovarian cancer, autoimmune diseases, malignant ascites), CD19 CD20 (B-cell malignancies), CD20 (non-Hodgkin's lymphoma), CD22 (leukemia, lymphoma, multiple myeloma, SLE), CD30 (Hodgkin's lymphoma), CD33 (leukemia, autoimmune diseases), CD38 (multiple myeloma), CD40 (lymphoma, multiple myeloma, leukemia (CLL)), CD51 (metastatic melanoma, sarcoma), CD52 (leukemia), CD56 (small cell lung cancer, ovarian cancer, meckel's cell carcinoma, and liquid tumors, multiple myeloma), CD66e (various cancers), CD70 (metastatic renal cell carcinoma and non-Hodgkin's lymphoma), CD74 (multiple myeloma), CD80 (lymphoma), CD98 (various cancers), mucin (various cancers), CD221 (solid tumors), CD227 (breast cancer, ovarian cancer), CD262 (Non-small cell lung cancer and other cancers), CD309 (ovarian cancer), CD326 (solid tumors), CEACAM3 (colorectal cancer, gastric cancer), CEACAM5 (carcinoembryonic antigen, CEA, CD66e) (breast, colorectal cancer, and lung cancer), DLL4 (Δ-like-4), EGFR (epidermal growth factor receptor, various cancers), CTLA4 (melanoma), CXCR4 (CD184, hematologic malignancies, solid tumors), Endoglin (CD105, solid tumors), EPCAM (epithelial cell adhesion molecule, bladder cancer, head and neck cancer, colon cancer, NHL prostate cancer, ovarian cancer), ERBB2 (epidermal growth factor receptor 2, lung cancer, breast cancer, prostate cancer), FCGR1 (autoimmune diseases), FOLR (folate receptor, ovarian cancer), GD2 gangliosides (various cancers), G-28 (cell surface antigen glycolipids, melanoma), GD3 idiotypes (various cancers), heat shock proteins (various cancers), HER1 (lung cancer, gastric cancer), HER2 (breast cancer, lung cancer, and ovarian cancer), HLA-DR10 (NHL), HLA-DRB(NHL, B-cell leukemia), human chorionic gonadotropin (various cancers), IGF1R (insulin-like growth factor 1 receptor, solid tumors, blood cancers), IL-2 receptor (interleukin 2 receptor, T-cell leukemia and lymphoma), IL-6R (interleukin 6 receptor, multiple myeloma, rheumatoid arthritis, Castleman's disease, interleukin 6-dependent tumors), integrin (αvβ3, α5β1, α6β4, α11β3, α5β5, αvβ5, various cancers), MAGE-1 (various cancers), MAGE-2 (various cancers), MAGE-3 (various cancers), MAGE-4 (various cancers), anti-transferrin receptor (various cancers), p97 (melanoma), MS4A1 (transmembrane 4-domain subfamily A member 1, non-Hodgkin's B-cell lymphoma, leukemia), MUC1 or MUC1-KLH (breast cancer, ovarian cancer, cervical cancer, bronchial cancer, and gastrointestinal cancers), MUC16 (CA125) (ovarian cancer), CEA (colorectal cancer), gp100 (melanoma), MART1 (melanoma), MPG (melanoma), MS4A1 (transmembrane 4-domain subfamily A member, small cell lung cancer, NHL), Nucleolin, Neu oncogene product (each cancer), P21 (various cancers), anti-(N-hydroxyacetylneuraminic acid) antibody binding site (breast cancer, melanoma), PLA-like testicular alkaline phosphatase (ovarian cancer, testicular cancer), PSMA (prostate cancer), PSA (prostate cancer), ROBO4, TAG 72 (tumor-associated glycoprotein 72, AML, gastric cancer, colorectal cancer, ovarian cancer), T-cell transmembrane protein (various cancers), Tie (CD202b), TNFRSF10B (tumor necrosis factor receptor superfamily member 10B, various cancers), TNFRSF13B (tumor necrosis factor receptor superfamily member 13B, multiple myeloma, NHL, other cancers, RA and SLE). TPBG (trophoblast glycoprotein, renal cell carcinoma), TRAIL-R1 (TNF-associated necrosis-inducing ligand receptor 1, lymphoma, NHL, colorectal cancer, lung cancer), VCAM-1 (CD106, melanoma), VEGF, VEGF-A, VEGF-2 (CD309) (various cancers). Other tumor-associated antigens that can be recognized by antibodies have been summarized and reviewed (Gerber, et al., mAbs 2009, 1:3, 247-253; Novellino, et al., Cancer Immunol Immunother. 2005, 54(3), 187-207; Franke, et al., Cancer BiotherRadiopharm. 2000, 15, 459-76. Examples of these antigens include: many other differentiation clusters (CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11a, CD11b, CD11c, CD12w, CD14, CD15, CD16, CDw17, CD18, CD21, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD31, CD32, CD34, CD35, CD36, CD37, CD41, CD42, CD43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49c, CD53, CD54, CD55, CD58, CD59, CD61, CD62E, CD62L, CD... 62P, CD63, CD68, CD69, CD71, CD72, CD79, CD81, CD82, CD83, CD86, CD87, CD88, CD89, CD90, CD91, CD95, CD96, CD100, CD103, CD105, CD106, CD109, CD117, CD120, D127, CD133, CD134, CD135, CD138, CD141, CD142, CD143, CD144, CD147, CD151, CD152, CD154, CD156, CD158, CD163, CD166, .CD168, CD184, CDw186, CD195, CD202 (a, b), CD209, CD235a, CD271, CD303, CD304), Annexin A1, Nucleolin, Endothelial glycoprotein (CD105), ROBO4, Aminopeptidase N, Δ-like 4 (DLL4), VEGFR-2 (CD309), CXCR49, CD184), Tie2, B7-H3, WT1, MUC1, LMP2, HPV E6 E7, EGFRvIII, EGFR, HER-2 / neu, Idiotype, MAGE A3, P53 nonmutant, NY-ESO-1, GD2, CEA, MelanA / MART1, Ras mutation, gp100, p53 mutant, Proteinase 3 (PR1), bcr-abl, Tyrosinase, Survivin, hTERT, Sarcoma translocation breakpoints, EphA2, PAP, ML-IAP, AFP, EpCAM, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, ALK, and AndrogenReceptor, Cyclin B1, Polysialic Acid, MYCN, RhoC, TRP-2, GD3, Fucosyl GM1, Mesothelin, PSCA, MAGE A1, sLe(a), CYP1B1, PLAC1, GM3, BORIS, Tn, GloboH, ETV6-AML, NY-BR-1, RGS5, SART3, STn, Carbonic Anhydrase IX, PAX5, OY-TES1, Spermin 17, LCK, HMWMAA, AKAP-4, SSX2, XAGE 1, B7H3, Pod Protein, Tie 2, Page4, VEGFR2, MAD-CT-1, FAP, PDGFR-β, MAD-CT-2, Fos Protein-Associated Antigen 1.

[0120] In another specific embodiment, the camptothecin analogue of this patent application coupled with a cell-binding molecule is used in compositions and methods for treating cancer. These cancers include, but are not limited to, adrenocortical carcinoma, anal cancer, bladder cancer, brain tumors (brainstem glioma, cerebellar astrocytoma, encephalomyeloma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal and pineal tumors, visual pathway and hypothalamic glioma), breast cancer, carcinoid tumors, gastrointestinal cancers, unknown small cell carcinoma, cervical cancer, colon cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family tumors (PNET), intracranial germ cell tumors, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer, gonadal genital cell tumors, gestational trophoblastoma, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, kidney cancer (renal cell carcinoma), laryngeal cancer, leukemia (acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic granulocytic, hair cell), lip and oral cancer, liver cancer, lung cancer (non-small cell, small cell), lymphoma (AIDS-related, central nervous system, skin T cell, Hodgkin's). Diseases (non-Hodgkin's disease), malignant mesothelioma, melanoma, Merkel cell carcinoma, metastatic squamous neck cancer and occult primary cancer, multiple myeloma and other plasma cell tumors, mycosis fungoides, myelodysplastic syndrome, myelodysplastic syndrome, nasopharyngeal carcinoma, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer (epithelial, germ cell tumor, low-grade tumor), pancreatic cancer (exocrine, islet cell carcinoma), paranasal sinus and nasal cavity cancer, parathyroid carcinoma, penile cancer Pheochromocytoma, pituitary tumors, plasma cell tumors, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma (renal cancer), renal pelvis and ureter (transitional cell), salivary gland cancer, Cesarean syndrome, skin cancer (cutaneous T-cell lymphoma, Kaposi's sarcoma, melanoma), small bowel tumors, soft tissue sarcomas, gastric cancer, testicular cancer, thymoma (malignant), thyroid cancer, urethral cancer, uterine cancer, unusual juvenile cancers, vaginal tumors, vulvar tumors, and Wilms' tumor.

[0121] In another specific embodiment, the conjugate of the camptothecin analogue and cell-binding molecule of this patent application is used in compositions and methods for treating or preventing autoimmune diseases. Autoimmune diseases include, but are not limited to, Achlorhydria, autoimmune active chronic hepatitis, acute disseminated encephalomyelitis, acute hemorrhagic leukoencephalitis, Addison's disease, azoospermia, alopecia areata, amyotrophic lateral sclerosis, ankylosing spondylitis, anti-GBM / TBM nephritis, antiphospholipid syndrome, anti-abnormal enzyme syndrome, arthritis, atopic allergy, atopic dermatitis, autoimmune aplastic anemia, autoimmune cardiomyopathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune peripheral neuropathy, autoimmune pancreatitis, autoimmune polyendocrine syndromes I, II, and III, autoimmune progesterone dermatitis, autoimmune thrombocytopenic purpura, autoimmune uveitis, Balo's disease / Balo concentric sclerosis, Bechets syndrome, Berger's disease, Bickerstaff encephalitis, Blau syndrome, bullous pemphigoid, Castleman's disease, Chagas disease, chronic fatigue immune dysfunction syndrome, chronic inflammatory demyelinating polyneuropathy, chronic relapsing multifocal osteomyelitis, chronic Lyme disease, chronic obstructive pulmonary disease, Churg-Strauss syndrome, cicatricial pemphigoid, celiac disease, Cogan syndrome, cold agglutinin disease, complement component 2 deficiency, craniotabes, CREST syndrome, Crohn's disease (idiopathic inflammatory bowel disease), Cushing's syndrome, cutaneous leukocytosis, Dego's disease, Dercum's disease, herpetic dermatitis, dermatomyositis, type 1 diabetes, diffuse systemic cutaneous sclerosis, Dressler's syndrome, discoid lupus erythematosus, eczema, endometriosis, enthesitis-associated arthritis, eosinophilic fasciitis, bullous pemphigoid Epidermolysis nodosum, erythema nodosum, idiopathic mixed cryoglobulinemia, Evans syndrome, fibrodysplasia ossification, fibromyalgia, fibrotic myositis, fibrosing alveolitis, gastritis, gastrointestinal pemphigoid, giant cell arteritis, glomerulonephritis, Goodpassulon syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, hemolytic anemia, allergic purpura, hepatitis of pregnancy, hidradenitis suppurativa, Hughes syndrome (antiphospholipid syndrome), hypogammaglobulinemia Idiopathic inflammatory demyelinating disease, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (autoimmune thrombocytopenic purpura), IgA nephropathy (Berge's disease), inclusion body myositis, inflammatory demyelinating polyneuritis, interstitial cystitis, irritable bowel syndrome, juvenile idiopathic arthritis, juvenile rheumatoid arthritis, Kawasaki disease, Langone-Eton myasthenic syndrome, leukocytic desquamative vasculitis, lichen planus, sclerosis, linear IgA disease (LAD), Lou Gehrig's disease (also known as amyotrophic lateral sclerosis), lupus hepatitis, lupus erythematosus, Majeed syndrome, Meniere's disease, microscopic polyarteritis, Miller-Fischer syndrome, mixed connective tissue disease, scleroderma, Muhammad-Haberman disease, Macaulay syndrome, multiple myeloma, multiple sclerosis, myasthenia gravis, myositis, somnolencephaly, neuromyelitis optica (Devic's disease), neuromuscular rigidity, eyelid cicatricial pemphigoid, Opsoclonus myoclonus syndrome, Ord thyroiditis, palindromic rheumatism, PANDAS (streptococcal-associated childhood autoimmune neuropsychiatric disorder), paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria, ParryRomberg syndrome, Parsonnage-Turner syndrome, flat cyclitis, pemphigus, pemphigus vulgaris, anemia, peripheral encephalomyelitis, POEMS syndrome, polyarteritis nodosa, polymyalgia rheumatica, polymyositis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive inflammatory neuropathy, psoriasis, psoriatic arthritis, gangrenous dermatitis, pure red cell aplasia, Rasmussen encephalitis, Raynaud's phenomenon, relapsing polychondritis, Reiter's syndrome, restless legs syndrome, postneurofibrosis, rheumatoid arthritis, rheumatoid fever, sarcoidosis, schizophrenia, Schmidt syndrome Comorbidities, Schnitzler syndrome, Schnitzler syndrome, scleritis, scleroderma, Sjögren's syndrome, spondyloarthritis, viscous blood disorders, Still's disease, stiff-person syndrome, subacute bacterial endocarditis, Sussac syndrome, Sweet syndrome, Sydenham's chorea, sympathetic anemia, Takayasu arteritis, temporal arteritis (giant cell arteritis), Tolosa-Hunt syndrome, transverse myelitis, ulcerative colitis (idiopathic inflammatory bowel disease), undifferentiated connective tissue disease, undifferentiated spondyloarthritis, vasculitis, vitiligo, Wegener's granulomatosis, Wilson's syndrome, and Wescott-Aldrich syndrome.

[0122] In another specific embodiment, the binding molecules on the conjugate for the treatment or prevention of autoimmune diseases include, but are not limited to, anti-elastin antibodies, Abys anti-epithelial cell antibodies, anti-basement membrane type IV collagen antibodies, antinuclear antibodies, anti-dsDNA, anti-ssDNA, anticardiolipin antibodies IgM, IgG, anti-celiac disease antibodies, antiphospholipid antibodies IgK, IgG, anti-SM antibodies, anti-mitochondrial antibodies, thyroid antibodies, microsomal antibodies, T-cell antibodies, thyroglobulin antibodies, anti-SCL-70, anti-Jo, anti-U.sub.1RNP, anti-La / SSB, anti-SSA, anti-SSB, anti-parietal cell antibodies, anti-histone, anti-RNP, C-ANCA, P-ANCA, anti-centromere, anti-fibrinogen, anti-GBM antibodies, anti-ganglioside antibodies, and anti-Desmogein. 3. Antibodies, anti-p62 antibody, anti-sp100 antibody, anti-mitochondrial (M2) antibody, rheumatoid factor antibody, anti-MCV antibody, anti-topoisomerase antibody, and anti-neutrophil cytoplasmic (cANCA) antibody.

[0123] In some preferred embodiments, the binding molecules on the conjugate of this patent can bind to receptors or receptor complexes expressed on initiating lymphocytes associated with autoimmune diseases. The receptors or receptor complexes include members of the immunoglobulin gene superfamily (e.g., CD2, CD3, CD4, CD8, CD19, CD20, CD22, CD28, CD79, CD90, CD152 / CTLA-4, PD-1, or ICOS), members of the TNF receptor superfamily (e.g., CD27, CD40, CD95 / Fas, CD134 / OX40, CD137 / 4-1BB, INF-R1, TNFR-2, RANK, TACI, BCMA, osteoprotein, Apo2 / TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4, and APO-3), integrins, cytokine receptors, chemokine receptors, major tissue compatibility proteins, lectins (C, S, or I), or complement control proteins.

[0124] In another specific embodiment, the available binding ligands with immune specificity to viral or microbial antigens are humanized or human monoclonal antibodies. "Viral antigens" include, but are not limited to, any viral peptides, polypeptides, or proteins capable of evoking an immune response (e.g., HIV gp120, HIV nef, RSV F glycoprotein, influenza virus neuraminidase, influenza virus hemagglutinin, HTLV Tax, herpes simplex virus glycoproteins (e.g., gB, gC, gD, and gE), and hepatitis B surface antigen). "Microbial antigens" include, but are not limited to, any microbial peptides, polypeptides, proteins, sugars, polysaccharides, or lipid molecules capable of evoking an immune response (e.g., bacterial, fungal, pathogenic protozoan, or yeast polypeptides, including, for example, LPS and capsular polysaccharide 5 / 8). Examples of antibodies that can be used to treat viral or microbial infections include, but are not limited to: palizumab, a humanized anti-respiratory syncytial virus monoclonal antibody used to treat RSV infection; PRO542, a CD4 fusion antibody used to treat HIV infection; oseltamivir, a human antibody used to treat hepatitis B virus; PROTVIR, a humanized IgG1 antibody used to treat cytomegalovirus; and an anti-LPS antibody.

[0125] The cell-binding molecule-camptothecin analogue conjugate described in this patent application can be used to treat infectious diseases. These infectious diseases include, but are not limited to, Acinetobacter infections, actinomycosis, African sleeping sickness (African trypanosomiasis), AIDS (Acquired Immunodeficiency Syndrome), amoebiasis, anaplasmosis, anthrax, hemolytic Yersinia infection, Argentine hemorrhagic fever, ascariasis, aspergillosis, astrovirus infection, babesiosis, Bacillus cereus infection, bacterial pneumonia, bacterial vaginosis, Bacteroidetes infection, giardiasis, ascariasis, BK virus infection, black hair nodule disease, human blastomycosis, blastomycosis, Bolivian hemorrhagic fever, spirochetal infection, botulism (and infant botulism), Brazilian hemorrhagic fever, brucellosis, Burkholderia infection, brucellosis ulcer, calicivirus infection (Nocturia cirrhosa). Such as viruses and Sapovirus), Campylobacteriosis, Candidiasis (candidiasis, thrush), Cat Scratch Disease, Cellulitis, Chagas Disease (American Trypanosomiasis), Ascositis, Chickenpox, Chlamydia, Chlamydia pneumoniae infection, Cholera, Melanoblastoma, Clonorchis sinensis, Clostridium difficile infection, Coccidioidomycosis, Colorado tick fever, Common cold (acute viral nasopharyngitis, acute rhinitis), Creutzfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, Cryptococcosis, Cryptosporidiosis, Cutaneous larval migration, Cyclosporidiosis, Enterobacterial infection, Enterovirus infection, Epidemic typhus, Erythema infectiosum (the fifth disease), Roseola, Fasciolopsis buski, Liver fluke disease, Fatal familial insomnia, Filariasis Clostridium perfringens food poisoning, live free amoeba infection, fusobacterium infection, gas gangrene (clostridium myonecrosis), dystrichomoniasis, Gerstmann-Strasseler-Schaker syndrome, Giardiasis, glanders, gonorrhea, granulomatous diarrhea (the fifth sexually transmitted disease), group A streptococcal infection, group B streptococcal infection, Haemophilus influenzae infection, hand-foot-and-mouth disease (HFMD), Hantavirus pulmonary syndrome, Helicobacter pylori infection, hemolytic uremic syndrome, hemorrhagic fever with renal syndrome, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, herpes simplex, histoplasmosis, hookworm infection, human bocavirus infection, human erythrozoonosis, human granulocytic anaplasmosis. Human metapneumovirus infection, human monocytic erythrocyte infection, human papillomavirus infection, human parainfluenza virus infection, hymenolepis taeniasis, Ebola virus infectious mononucleosis, influenza, isosporidiosis, Kawasaki disease, keratitis, Agrimonia pilosa infection, kuru, Lassa fever, Legionnaires' disease, Pityak fever, leishmaniasis, Lyme disease, lymphatic filariasis (elephantiasis), lymphocytic choriomeningitis, malaria, Marluburg hemorrhagic fever, measles, glanders (Wyeth's disease), meningitis, meningococcal infection, metaclonorchiasis, microsporidiosis, molluscum contagiosum, mumps, mouse typhus (endemic typhus), mycoplasmal pneumonia, mycoplasma gondii, myiasis.Neonatal conjunctivitis (neonatal eye disease), variant Creutzfeldt-Jakob disease (vCJD, nvCJD), nocardiac infection, onchocerciasis (river blindness), paracoccidioidomycosis (South American blastomycosis), paragonimiasis, pastoral infection, head lice, body lice, pubic lice, pelvic inflammatory disease, pertussis, plague, pneumococcal infection, Pneumocystis pneumonia, pneumonia, poliomyelitis, Prevotella infection, primary amoebic meningoencephalitis, progressive multifocal leukoencephalopathy, psittacosis, Q fever, rabies, rat-bite fever, respiratory syncytial virus infection, rhinocystis infection, rhinovirus infection, rickettsial infection, rickettsial pox, Rift Valley fever, Rocky Mountain spotted fever, rotavirus infection, rubella, salmonellosis, SARS (Severe Acute Respiratory Syndrome), scabies, schistosomiasis, sepsis, Shigella. Bacillary dysentery, herpes zoster, smallpox, sporothrix, staphylococcal food poisoning, Staphylococcus aureus infection, strongyloidiasis, syphilis, tapeworm infection, tetanus, tinea barbae, tinea scalp, tinea corporis, tinea cruris, tinea manuum, palmar-ulcerative tinea, tinea pedis, onychomycosis, tinea versicolor, toxocariasis (ocular larval migration syndrome), toxoplasmosis, trichinosis, trichomoniasis, whipworm infection, tuberculosis, tularemia, ureaplasma infection, Venezuelan equine encephalitis, Venezuelan hemorrhagic fever, viral pneumonia, West Nile fever, tinea pedis (white tinea), Yersinia pseudotuberculosis, Yersinia pestis enteropathy, yellow fever, zygomycosis.

[0126] The cell-binding agent of the present invention is preferably an antibody, and the pathogenic strains it antagonizes include, but are not limited to, Acinetobacter baumannii, Actinobacter spp., Actinobacteria and Propionibacterium acnes, Trypanosoma brucei, HIV (human immunodeficiency virus), Entamoeba histolytica, Anaplasma, Bacillus anthracis, Vibrio hemolyticus, Junin virus, Ascaris spp., Aspergillus spp., Astroviridae, Babesia spp., Bacillus cereus, various bacteria, Bacteroides spp., Escherichia coli, Ascaris spp., BK virus, and Nodularia spp.* *Blastocystis humanis*, *Blastobacterium dermatitidis*, Machubo virus, *Bretorius*, *Clostridium botulinum*, *Clematis*, *Brucella*, usually *Burkholderia cepacia* and other *Burkholderia* species, *Mycobacterium ulcerans*, *California*, *Campylobacter*, usually *Candida albicans* and other *Candida* species, *Bartonella henselae*, Group A Streptococcus and Staphylococcus, *Trypanosoma krusei*, *Haemophilus ducreyi*, varicella-zoster virus (VZV), *Chlamydia trachomatis*, Colorado tick-borne fever virus, rhinovirus, coronavirus, CJD prions, Crimean-Congo hemorrhagic fever virus, *Cryptococcus neoformans*, *Cryptosporidium*, *Hookworm brasiliensis*, various parasites, *Cyclospora*, *Taenia zoster*, cytomegalovirus, dengue virus (DEN-1). DEN-2, DEN-3, and DEN-4) - Flaviviritis, Bifidobacterium fragilis, Corynebacterium diphtheriae, Sparganum, Draconis medina, Ebola virus, Echinococcus spp., Erickettsia spp., Enterococcus spp., Enterovirus spp., Rickettsia prowleri, Parvovirus B19, Human herpesvirus 6 and Human herpesvirus 7, Fasciolopsis buski, Fasciolopsis buski and Fasciola gigantica, FFI prions, Filariaceae superfamily, Clostridium perfringens, Fusobacterium spp., other Clostridium spp., Geotrichum candida, GSS prions, Giardia lamblia, Burkholderia, Microsporum spp. and Candida graminearum, Neisseria gonorrhoeae, Klebsiella granulomatosa, Streptococcus pyogenes, Streptococcus agalactiae, Haemophilus influenzae, Enteroviruses, mainly Coxsackieviruses. Odd A virus and enterovirus 71, Sinopharm virus, Helicobacter pylori, Escherichia coli O157:H7, Bunyaviridae, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Hepatitis D virus, Hepatitis E virus, Herpes simplex virus 1, Herpes simplex virus 2, Histoplasma capsulatum, Haemophilus influenzae (duodenal adenoma and ampullary carcinoma), human bocavirus, Ehrlichella, Haemophilus aphagophilus, human metapneumovirus, Ehrlichella chaffi, human papillomavirus, human parainfluenza virus, Hymenolepis microspinipes and Hymenolepis atrophicus, Epavirus, Orthomyxoviridae family, Isococcus benzii, Agrobacterium, Klebsiella pneumoniae, Klebsiella pneumoniae, Legionella pneumophila, Legionella pneumophila, Legionella pneumophila Leishmania, Mycobacterium leprae and Mycobacterium tuberculosis, Leptospira, Listeria monocytogenes, Borrelia burgdorferi and other Borrelia species, Trichinella bancrofti and Filaria malayi, Lymphocytic choriomeningitis virus (LCMV), Plasmodium, Marluburg virus, Measles virus, Burkholderia melioides, Neisseria meningitidis, Paragonimus yokokawa, Microsporidioides, Molluscum contagiosum virus (MCV), Mumps virus, Rickettsia typhi, Mycoplasma pneumoniae, various bacteria and fungi parasitizing Dipterocarp larvae, Chlamydia trachomatis and Neisseria gonorrhoeae, vCJD prions, Nocardia and other Nocardia species, Onchocerca, Oncocerca subfamily, Parasaurolophus, Cimania and other paragenera.Pasteurella spp., head lice, human lice, Bodotriella pertussis, Yersinia pestis, Streptococcus pneumoniae, Streptococcus pneumoniae, poliovirus, Prevotella spp., Neisseria gonorrhoeae, JC virus, Chlamydia psittaci, Coxella burgdorferi, rabies virus, monostreptococci and spirochetes, respiratory syncytial virus, rhinosporidia, rhinovirus, Rickettsia spp., Rickettsia microsporum, Rift Valley fever virus, Rickettsia rickettsiae, rotavirus, rubella, Salmonella spp., SARS Coronavirus, human scabies mite, Schistosoma spp., Somatic cell spp., Shigella spp., varicella-zoster virus, smallpox or pustular smallpox, Sporothrix schenckii, Staphylococcus aureus spp., Staphylococcus aureus, Streptococcus pyogenes, Strongyloides, Treponema pallidum, Tapeworm spp., Tetanus, Trichophyton spp., Trichophyton spp., Epidermophyton floccosum, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton rubrum, Epidermophyton vinifera, Trichophyton spp., Trichophyton spp., Cell death spp., Arrowhead Poison or toxoplasmosis, Toxoplasma gondii, Trichinella spiralis, Trichomonas vaginalis, Trichomonas vaginalis, Mycobacterium tuberculosis, Francisella foetida, urea and equine encephalitis virus, Venezuelan equine encephalitis virus, Vibrio cholerae, Guanarito virus, West Nile virus, Saccharomyces basilaria, Yersinia pseudotuberculosis, Yersinia enterocolitica, yellow fever virus, Mucorales (Mucor fungal infections) and Entomophthora (Entomycosis fungal infections), Mucorales Pseudomonas aeruginosa, Campylobacter (Vibrio), Aeromonas, Escherichia coli, Yersinia, Shigella dysenteriae, Shigella, Shigella, Salmonella, Salmonella typhi, Treponema pallidum, Treponema vinsenii, Borrelia burgdorferi, Treponema pallidum, Pneumocystis carinii, Brucella abortus, Brucella, Mycoplasma, Rickettsia prowleri, Rickettsia tsutsugamushi, Chlamydia, Pathogenic fungi (Aspergillus fumigatus, Candida albicans, Histoplasma capsulatum, protozoa (Entamoeba histolytica, Trichomonas tenas, Trichomonas hominis, Trypanosoma gambiae, Trypanosoma rhodesianum, Leishmania rhodesii, Leishmania tropicalis, Leishmania brasiliensis, Pneumocystis pneumonia, Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae) or Helminiths (Schistosoma japonicum, Schistosoma mansoni, Schistosoma haematobium, and hookworm).

[0127] The cell binding agent used in this patent application, and other antibodies for the treatment of viral diseases, including but not limited to antibodies against the following pathogenic viral antigens: poxvirus, herpesvirus, adenovirus, flavivirus, enterovirus, picornavirus, parvovirus, reovirus, retrovirus, influenza virus, parainfluenza virus, mumps, measles, respiratory syncytial virus, rubella, arbovirus, rhabdovirus, Salmonella, non-A / non-B hepatitis virus, rhinovirus, coronavirus, Rotor virus; oncogenic viruses such as HBV (hepatocellular carcinoma), human papillomavirus (HPV) (cervical cancer, anal cancer), Kaposi's sarcoma-associated herpesvirus (Kaposi's sarcoma), human herpesvirus type 4 (nasopharyngeal carcinoma, Burkitt lymphoma, primary central nervous system lymphoma), MCPyV (Merkel cell carcinoma), SV40 (simian virus 40), HCV (hepatocellular carcinoma), HTLV-1. (Adult T-cell leukemia / lymphoma); viral infections caused by immune disorders, such as human immunodeficiency virus (AIDS); viruses affecting the central nervous system, such as JCV (progressive multifocal leukoencephalopathy), hepatitis C virus MeV (subacute sclerosing panencephalitis), LCV (lymphocytic choroid plexus meningitis), bronchodilator encephalitis, orthomyxovirus (encephalitis), RV (rabies), proboscis virus, herpesvirus meningitis, Ramsey Hunt syndrome type II, poliovirus (poliomyelitis virus, postpoliomyelitis syndrome), HTLV-1 (tropical paralytic leukemia). Paralysis); Cytomegalovirus (CMV) retinitis, HSV (herpetic keratitis); cardiovascular viruses, such as CBV (pericarditis, myocarditis); respiratory system / acute viral rhinitis / viral pneumonia viruses, such as Epstein-Barr virus (EBV infection / infectious mononucleosis), cytomegalovirus, SARS coronavirus (severe acute respiratory syndrome) or orthomyxovirus, influenza A / B / C (influenza / avian influenza), paramyxovirus, human parainfluenza virus, RSV (human respiratory syncytial virus), hMPV; digestive system viruses, such as (MuV) (Mumps) mumps virus, cytomegalovirus (cytomegalovirus esophagitis), adenovirus (adenovirus infection), rotavirus, norovirus, astrovirus, coronavirus, HBV (hepatitis B virus), CBV, HAV (hepatitis A virus), HCV (hepatitis C virus), HDV (hepatitis D virus), HEV (hepatitis E virus), HGV (hepatitis G virus); urogenital viruses, such as BK virus and MuV virus (mumps).

[0128] Furthermore, the present invention also includes the conjugates and acceptable carriers of the present invention for the treatment of cancer and autoimmune diseases. Methods for treating cancer and autoimmune diseases can be performed in vitro, in vivo, or ex vivo. Examples of in vitro use include treating cell cultures to kill all cells except variants that do not express the target antigen; or to kill variants that express unwanted antigens. Examples of ex vivo use include treating hematopoietic stem cells (HSCs) prior to transplantation (HSCT) to kill diseased or malignant tumor cells. For example, removing tumor cells or lymphocytes from bone marrow prior to autologous transplantation in cancer treatment or in the treatment of autoimmune diseases, or removing T cells and other lymphocytes from allogeneic bone marrow or tissue prior to transplantation to prevent graft-versus-host disease. Such clinical ex vivo treatment can be performed by harvesting bone marrow from a patient or other individual and incubating it in a serum-containing culture medium at approximately 37 °C for approximately 15 minutes to approximately 48 hours, to which the conjugates of the present invention are added at concentrations ranging from approximately 1 pM to 0.1 mM. The specific drug concentration and incubation time (= dosage) should be determined by a qualified clinician. After incubation, the bone marrow cells are washed with serum-containing culture medium and administered intravenously to the patient according to known methods. If the patient receives other treatments (such as ablation chemotherapy or total body radiation therapy) between bone marrow collection and re-infusion of therapeutic cells, the treated bone marrow cells should be cryopreserved in liquid nitrogen using standard medical equipment.

[0129] Stable conjugates should “maintain their biological activity” in pharmaceutical formulations. For example, if the biological activity of the conjugate can be maintained within 20%, preferably 10% (within the measurement error), according to the methods of antigen binding assay and / or in vitro cytotoxicity assay, for a given period of time, such as 12 months.

[0130] For clinical in vivo use, the conjugate of the present invention will be provided in solution or lyophilized solid form, which can be dissolved in sterile water for injection. Suitable conjugate administration regimens are as follows: once daily, once weekly, once every two weeks, once every three weeks, once every four weeks, or once monthly for 8–54 weeks, intravenously. The injection dose is in 50–1000 mL of normal saline, optionally supplemented with human serum albumin (e.g., 0.5–1 mL of concentrated human serum albumin solution, 100 mg / mL). The weekly dose is approximately 50 µg to 20 mg / kg body weight, intravenously (in the range of 10 µg to 200 mg / kg per injection). Patients may receive a second course of treatment 4–54 weeks after initial treatment. Specific clinical protocols regarding administration methods, excipients, diluents, dosage, timing, etc., can be determined by a skilled clinician.

[0131] Examples of diseases that can be treated by killing specific cells in vivo or in vitro include any type of malignancy, autoimmune diseases, transplant rejection, and infectious diseases (viruses, bacteria, or parasites).

[0132] The amount of conjugate required to achieve a satisfactory biological effect will depend on many factors, including the chemical properties of the conjugate, its potency and bioavailability, the type of disease, the patient's ethnicity, the patient's disease state, and the route of administration, which determine the required dose, manner of administration, and administration regimen.

[0133] Generally, the drug comprising the present invention can be administered parenterally as a physiologically buffered aqueous solution containing 0.1 to 10% w / v of the conjugate. Typical dosage ranges are 1 µg / kg to 0.1 g / kg body weight once daily, weekly, bi-weekly, bi-weekly, or monthly, with preferred dosage ranges being human equivalent doses of 0.01 mg / kg to 20 mg / kg body weight once weekly, bi-weekly, bi-weekly, or monthly. The preferred dosage of the drug to be administered may depend on variables such as the type of disease or disease progression, the patient's overall health status, the relative biological efficacy of the selected compound, the drug formulation, the route of administration (intravenous, intramuscular, or other), the pharmacokinetic properties of the drug along the defined route of administration, the rate of administration (bolus or continuous infusion), and the administration regimen (number of repetitions within a given time).

[0134] The conjugates of the present invention can also be administered in unit dose form, wherein the term "unit dose" refers to a single dose that can be given to a patient and can be easily disposed of and packaged, while the active conjugate itself or the pharmaceutically acceptable composition described below maintains physical and chemical stability at the unit dose. Typical daily / weekly / bi-weekly / monthly total dose ranges from 0.01 to 100 mg / kg body weight. As a general guideline, the human unit dose range is 1 mg to 3000 mg daily / weekly / bi-weekly / tri-weekly / monthly. A preferred unit dose is 1 to 500 mg, once to four times a month, more preferably 1 mg to 100 mg, once weekly / bi-weekly / tri-weekly. The conjugates provided herein can be formulated into pharmaceutical compositions by mixing with one or more pharmaceutically acceptable excipients. Such unit-dose compositions can be administered orally, as a drug in the form of tablets, simple capsules, or soft gel capsules; or intranasally, as a drug in the form of powders, nasal drops, or aerosols; or by skin administration, such as using topical ointments, creams, lotions, gels, or sprays, or by transdermal patches. The compositions can be conveniently administered in unit dosage forms and can be prepared by methods generally known in the pharmaceutical field, such as those described in Remington: The Science and Practice of Pharmacy, 21 thed.; Lippincott Williams & Wilkins: Philadelphia, PA, 2005.

[0135] Preferred formulations include pharmaceutical compositions, preferably administered orally or enterally. For oral dosage forms, such as lozenges, powders, capsules, and tablets, one or more of the following components or other compounds with similar properties may be included: binders, such as microcrystalline cellulose or astragalus gum; diluents, such as starch or lactose; dispersants, such as starch and cellulose derivatives; lubricants, such as magnesium stearate; glidants, such as silica colloids; sweeteners, such as sucrose or saccharin; and flavor enhancers, such as menthol or methyl salicylate. Capsules can be in hard or soft capsule form and are generally made from gelatin mixtures or mixed with plasticizers, as are starch capsules. Additionally, the unit dosage form may contain various other ingredients, altering their physical form, such as sugar coating, shellac, or enteric solvents. Other oral dosage forms, such as syrups or elixirs, may contain sweeteners, preservatives, pigments, colorants, and flavorings. In addition, active compounds can be processed and formulated into rapidly soluble dosage forms, controlled-release dosage forms, or sustained-release formulations, with sustained-release formulations being the preferred option. Tablets are preferably formulated using combinations of lactose, corn starch, magnesium silicate, croscarmellose sodium, polyvinylpyrrolidone, magnesium stearate, talc, etc.

[0136] Parenteral liquid formulations include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Liquid formulations may also contain binders, buffers, preservatives, chelating agents, sweeteners, flavorings, and colorings, etc. Non-aqueous solvents include alcohols, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and organic lipids such as ethyl oleate. Aqueous solvents include mixtures of water, alcohols, buffers, and salts; in particular, biocompatible, biodegradable lactide polymers, lactide / glycolic acid polymers, or polyethylene glycol / polyglycerol copolymers can be used as excipients to control the release of the active drug. Excipients for intravenous administration may include liquid and nutritional supplements, electrolyte supplements, and Ringer's glucose-based excipients, as well as similar excipients. Other feasible parenteral delivery systems for the active drug of the present invention include ethylene-vinyl acetate copolymer microparticles, implantable osmotic pumps, and liposomes.

[0137] Other feasible delivery methods include inhalers, including dry powders, aerosols, and water drops. Inhalers can be solutions containing substances such as polyethylene glycol-9-lauryl ether, glycholates, deoxycholates, or oils, and can be administered as nasal drops or intranasal colloids. Oral medications include tablets and candy tablets, which may contain flavoring agents such as sucrose, gum arabic, and other excipients such as glycholates. Suppositories are suitable for single-dose formulations, using solids such as cocoa oil as a carrier, and may also include salicylic acid. Topical skin medications are preferably in the form of ointments, emulsions, lotions, patches, gels, sprays, aerosols, or oils. Petrolatum, lanolin, polyethylene glycol, alcohols, and mixtures thereof can be used as drug carriers. Skin delivery dosage forms can be patches, emulsions, buffer solutions, or solutions dissolved or dispersed in polymers or adhesives.

[0138] In another specific embodiment, the conjugate of the invention application can act in conjunction with other known or soon-to-be-known therapeutic agents, such as chemotherapy drugs, radiotherapy drugs, immunotherapy drugs, autoimmune disease drugs, anti-infective drugs, or other antibody drug conjugates, to achieve a synergistic effect. In another specific embodiment, the synergistic drug or radiotherapy can be administered or performed before or after the administration of the conjugate drug of the present invention. This can be done 1 hour, 12 hours, 1 day, 1 week, 2 weeks, 3 weeks, 1 month, or even several months before or after the administration of the conjugate drug of the present invention.

[0139] In other embodiments, the synergistic drugs include, but are not limited to: 1) Chemotherapy drugs: a) Alkylating agents, such as nitrogen mustard: clobenazine, cloprazosin, cyclophosphamide, dacarbazine, estradiol nitrogen mustard, ifosfamide, nitrogen mustard, dimethoxyamine hydrochloride, dioxane, amlodipine hydrochloride, mycophenolic acid, eugenol, guanobromide, neonitrogen mustard, benzyl mustard cholesterol, pine oxychloride, thiotetracycline, trofenamic acid, uracil; CC-1065 (including its adolaxine and carbendazim) New, pyrazinamide and their synthetic analogues); docalamycin (including KW-2189 and CBI-TMI, and their synthetic analogues); benzodiazepine dimers (e.g., pyrrole benzodiazepine (PBD) or tometamycin, indole benzodiazepine, imidazole benzothiazepine or oxazolidine benzodiazepine dimers); nitrosoureas (carmustine, lomustine, clostridium chloride, formustine, nimodipine) a) Lamustin; alkyl sulfonates (sulphurein, sulphurein, sulfamethoxazole and pisufen); triazine (dacarbazine); platinum-containing compounds (carboplatin, cisplatin, oxaliplatin); aproidines, such as benzodihydroguanidine, calcodone, methotrexate and uredopa; ethyleneimine and methylmelamine, including hexamethylmelamine, triethylenetriamine, triethylphosphatidine, triethylenethiophosphatidine and tris(hydroxymethyl)methylamine; b) plant alkaloids: such as vinca alkaloids (vincrine, vinblastine, vinorelbine, vinorelbine, norvincrine); paclitaxel derivatives (paclitaxel, docetaxel and their analogues); maytansine derivatives (DM1, DM2, DM3, DM4, maytansine, salinomycin and their analogues); cryptophycin (especially cryptophycin) 1) and cryptophycin 8); epothilone, soft coral alcohol, dimolide, bryophyll, sea haretoxin, auristatin, cephalostatin; pancratistatin; sarcodictyin; sponge inhibitor; c) DNA topoisomerase inhibitors, such as etoposide-tinib (9-aminocamptothecin, camptothecin, cristatin, doramycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, norsulfuron, retinoid (retinol), teniposide, topotecan, 9-nitrocamptothecin (RFS 2000)); mitomycin (mitomycin C); d) antimetabolites, such as antifolate agents, DHFR inhibitors (methotrexate, tromethamine, folate, pteroxate, aminopterin (4-aminobenzoic acid) or other folic acid analogs); IMP dehydrogenase inhibitors (mycophenolate mofetil, thiazofuran, ribavirin, EICAR); ribonucleotide reductase inhibitors (hydroxyurea, deferoxamine); pyrimidine analogs, uracil analogs (ancitabine, azacitidine, 6-azauracil, capecitabine (Xeloda), carmoflurane, cytarabine, dideoxyuridine, deoxyfluorouridine, enoxabin, 5-fluorouracil, fluorouridine, ratitrexed (Tomudex));Cytosine analogues (cytarabine, cytosine arabinoside, fludarabine); purine analogues (azathioprine, fludarabine, mercaptopurine, thiamine, thioguanine); folic acid supplements, such as florlinic acid; e) hormone therapy agents, such as receptor antagonists, anti-estrogens (medroxyprogesterone acetate, raloxifene, tamoxifen), LHRH agonists (gostalin, leuprolide acetate); anti-androgens (bicalutamide, flutamide, calusone, betahistelone propionate, epiandrosterone, goserelin, leuprolide). Lin, meteridone, nilumid, testosterone, tralostertan and other androgen inhibitors; retinoids, vitamin D3 analogs (CB1093, EB1089, KH1060, cholecalciferol, ergocalciferol); photodynamic therapy agents (verteporfin, phthalocyanine, photosensitizer Pc4, demethoxy-rubigin A); cytokines (interferon-α, interferon-γ, tumor necrosis factor (TNF), TNF-containing human proteins); f) kinase inhibitors, such as BIBW 2992 (anti-EGFR / Erb2), Imatinib, Gefitinib, Guagatanib, Sorafenib, Dasatinib, Sunitinib, Erlotinib, Nilotinib, Lapatinib, Axitinib, Pazopanib, Vandetanib, E7080 (anti-VEGFR2), Mubritinib, Ponatinib (AP24534), Bafetinib (INNO-406), Bosutinib (SKI-606), Cabozantinib, Vemoderil, Iniparib, Russoritinib, CYT387, Axitinib, Viozonab, Sorafenib, Bevacizumab, Cetuximab Trastuzumab, ranibizumab, panitumumab, ispinazine; g) Poly(ADP-ribose) polymerase (PARP) inhibitors, such as olaparib, niraparib, iniparib, tarazoparib, veliparib, veliparib, CEP 9722 (Cephalon), E7016 (Eisai), BGB-290 (Beigene), 3-aminobenzoxamide; h) Antibiotics, such as enediyne antibiotics (cachicin, especially cachicin γ1, δ1, α1 and β1 (ref. J. Med. Chem. 1996, 39(11), 2103-2117;Angew Chem Intl. Ed. Engl. 1994, 33:183-186), dyinmycin, including dyinmycin A and desoxymycin, esperamycin, katimycin, C-1027, madaropeptin, neocarcinoxine, oxaline and related chromogen-derived acetylenoid antibiotics), aclacinomysins, actinomycin, atrazomycin, diazoserine, Bleomycin, carnomycin, kanamycin, erythromycin, carcinogen, chromomycin, dachlormycin, daunorubicin, dedaunorubicin, 6-diazo-5-sideoxy-L-norleucine, doxorubicin, morpholine-doxorubicin, cyanomorpholine-doxorubicin, 2-pyrrolidone doxorubicin and deoxydaunorubicin, epirubicin, arubicin, idarubicin, macromycin, nitomycin, mycophenolic acid, nogamycin, olivomycin, peplomycin, potfiromycin, puromycin, queramycin, rodolmycin, streptomycin, streptozotocin, tuberculin, ubenimex, phenobarbital, zorubicin; i) others, such as polyketides (anthocyanins), especially bullatacin and bullatacinone; gemcitabine, cyclooxygenases (such as carfenomib), bortezonis Mitochondrial, Thalidomide, Lenalidomide, Pomalidomide, Tosedostat, Zybrestat, PLX4032, STA-9090, Stimuvax, Allovoctin-7, Xegeva, Provenge, Yervoy, Isoprenoidation inhibitors (e.g., Lovastatin), Dopaminergic neurotoxins (e.g., 1-methyl-4-phenylpyridinium ion), Cell cycle inhibitors (e.g., Astrococcus), Actinomycins (e.g., Actinomycin D, Dermatomycin), Bleomycin (e.g., Bleomycin A2, Bleomycin B2, Pelomycin), Anthracycline antibiotics (e.g., Daunorubicin), Amatoxins, Adriamycin, Idarubicin, Epirubicin, Pirarubicin, Zorobacterium, Mitoxantrone, MDR inhibitors (e.g., Verapamil), Ca 2+ ATPase inhibitors (such as carotenoids), histone deacetase inhibitors (vorinostat, romedixin, pabistal, valproic acid, mocetinostat (MGCD0103), Belinostat, PCI-24781, entenolide, SB939, resminostat, givinostat, AR-42, CUDC-101, sulforaphane, trachomatis A); celecoxib, glitazones, epigallocatechin gallate, disulfiram, salinosporamide A;Antiadrenergic drugs, such as aminoglutethimide, mitotane, tralostertan, glucuronolactone, aldoxime, aminoacetylpropionic acid, acridine, arabinoside, bestrabucil, bisacodyl, edatraxate, defofamine, mecosine, diazinon, efornithine (DFMO), elfomithine, elifonitrile, etoglucopyranoside, gallium nitrate, cytosine, hydroxyurea, ibandronate, lentinan, chlorpheniramine, mitoxantrone, mitoxantrone, mogutarol, diaminonitroazidine, pentostatin, methamidophos, pirarubicin, podophyllin, 2-ethylhydrazine, methylbenzylhydrazine; PSK ®; guanazin dionepropane; rhizomycin; cizole; spirocyclic germanium; succinate; triamine quinone; trichlorotriethylamine; trichothecenes (especially T-2 toxin, verrucosporin A, baculosporin A, and anguidine), polyurethanes, siRNA, antisense drugs, and nucleases; 2) Medications for autoimmune diseases, including but not limited to: cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine, chlorambucil, chloroquine, cyclophosphamide, corticosteroids (e.g., ancinonide, betamethasone, budesonide, hydrocortisone, flunisolone, fluticasone propionate, flucloxacillin, danazol, dexamethasone, triamcinolone, beclomethasone dipropionate), DHEA, etanercept, hydroxychloroquine, infliximab, meloxicam, methotrexate, mycophenolate mofetil, prednisone, sirolimus, tacrolimus; 3) Anti-infective agents include, but are not limited to: a) Aminoglycosides: amikacin, Wuyimycin, gentamicin (netilmicin, sisomicin, isapamicin), hygromycin, kanamycin (amikacin, abekacin, aminodeoxykanamycin, dibekacin, tobramycin), neomycin (neomycin B, paromomycin, ribosomycin), netilmicin, spectinomycin, streptomycin, tobramycin, methylsodium thiosulfate; b) Azamides: chloramphenicol, chloramphenicol, flumethinazole, methomyl; c) Ansarmycins: gerdemycin, chloramphenicol; d) Carbapenems: biapenem, doripenem, ertapenem, imipenem / cilastatin, meropenem, panipenem; e) Cephalosporins: Carbazocephalosporins (carbazocephalosporins), cefaclor, cefaclor, cefadroxil, cefarothiopin, cefotaxime, cefalothin or cephalosporins, cefalexin, cefamandole, cefepime, ceftriaxone, cefotaxime, cefoxitin, cefazolin, cefazolin, cefadroxil, cefacarine, cefodoxime, cefepime, cefminox, cefoxitin, cefprozil, cefazolin, cefotaxime, cefoperazone, cefoperazone, cefdextrin, cefepime Cefotaxime, cefotaxime, cefodizine, cefnicotinic acid, cefotiam, cefotaxime, cefotaxime, cefotiam, cefazolin, cefoperazone, cefepime, cefpirome, cefpirome, cefpodoxime, cefprozil, cefquinoxime, cefsulfuron-methyl, ceftazidime, cefoteran, cefbuprofen, cefotaxime, ceftriaxone, cefuroxime, cefazolin, cephalosporins (cefoxitin, cefotetan, cefmetazole), oxocephalosporins (fluoxocephalosporin, latamoxporin); f) Glycopeptides: Bleomycin, Vancomycin (Olivancin, Tervavancin), Teicoplanin (Dabavancin), Ramolane; g) Glycines: such as Tigecycline; h) β-lactamase inhibitors: Penicillane (Sulbactam, Tazobactam), Clavulanic acid; i) Lincosamides: Clindamycin, Lincomycin; j) Lipopeptides: Daptomycin, A54145, Calcium-dependent antibiotics (CDA); k) Macrolides: Azithromycin, Quinoerythromycin, Clarithromycin, Dierythromycin, Erythromycin, Fluerythromycin, Josamycin; Ketolides (Telithromycin, Quinoerythromycin), Midecamycin, Meokamycin, Rifampicin, Rifamycin (Rifampin, Rifampicin, Rifabutin, Rifapentine), Rotamycin, Roxithromycin, Spectinomycin, Spiramycin, Tacrolimus (FK506), Acetylosin, Telithromycin; l) Monocyclic β-lactam antibiotics: Aztreonam, Tegamona; m) Azoxystones: Linezolid; n)Penicillins: Amoxicillin, Ampicillin (Pipacillin, Hetacillin, Bambucillin, Metancillin, Phthalimicillin), Azidocillin, Aloxicillin, Penicillin, Benzathine Penicillin, Phenoxybenzyl Penicillin, Clomicillin, Procaine Penicillin, Carbenicillin (Carinin), O-cloxacillin, Dicloxacillin, Cefazolin, Flucloxacillin, Mecillin (Azamidin Diester), Meropenem, Methicillin, Nafcillin, Oxadiazine, Acetocillin, Penicillin, Necillin, Penicillin, Guaricillin, Phenprocillin, Sulbenicillin, Temoxicillin, Tekacillin; o). Polypeptides: Bacitracin, Polymyxin E, Polymyxin B; p). Quinolones: Araflavin, Barofloxacin, Ciprofloxacin, Clinfloxacin, Daflonax, Difloxacin, Enrofloxacin, Floxin, Garefloxacin, Gatifloxacin, Gemmifloxacin, Gaptafloxacin, Travafloxacin, Levofloxacin, Lomefloxacin, Mebofloxacin, Moxifloxacin, Naflufloxacin, Norfloxacin, Obibixin, Ofloxacin, Pefloxacin, Travafloxacin, Gaptafloxacin, Sitafloxacin, Sparfloxacin, Temafloxacin, Tosufloxacin, Travafloxacin; q). Antibiotics: Pannamycin, Quinupordin / Dalfopristin; r). Sulfonamides: Sulfamilone, Prontocillin, Sulfaacetic acid, Sulfamethoxazole, Sulfasalazine, Sulfamethoxazole, Trimethoprim- Sulfamethoxazole (compound sulfamethoxazole); s) Steroid antibiotics: such as fusidic acid; t) Tetracyclines: doxycycline, chlortetracycline, chlortetracycline, demeclocycline, lysine, chloramphenicol, methacycline, minocycline, oxytetracycline, penicillin tetracycline, rolicycline, tetracycline, glycine (such as tigecycline); u) Other types of antibiotics: annonaceous, arsenadamine, bacterial terpene inhibitors (bacitracin), DADAL / AR inhibitors (cycloserine), dictyostatin, spongolactone, ilexelose, epothilone, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole, mupirocin, mycolactones, NAM synthesis inhibitors (such as fosfomycin), nitrofurantoin, paclitaxel, acanthopanax, pyrazinamide, quinupristin / dalfopristin, rifampin, tazobactam-tinidazole, anechoic acid lactone; 4) Antiviral drugs: a) Entry / fusion inhibitors: apraviroc, maraviro, velivoro, gp41 inhibitor (enfvir), PRO140, CD4 inhibitor (ibalizumab); b) Integrase inhibitors: raltegravir, ertelavin, globoidnan A; c) Maturation inhibitors: bevirimiloma, vivecon; d) Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir; e) Nucleosides and nucleotides: abacavir, acyclovir, adefovir, amdoxovir, apricotibine, brivudine, cidofovir clavudine, dexavistatin, doxavistatin (DDI), apricotibine, emtricitabine (FTC), entecavir, famciclovir, fluorouracil (5-FU), 3'-fluorosubstituted 2',3'-dideoxynucleoside derivatives (e.g., 3'-fluoro-2',3'-dideoxythymidine (FLT) and 3'-fluoro-2 ',3'-Dideoxy (FLG) fomivir, ganciclovir, idoxuridine, lamivudine (3TC), L-nucleoside (such as β-L-thymidine, β-L-2'-deoxycytidine), penciclovir, racivir, ribavirin, stampidine, stavudine (d4T), tarivilin (veramidine), telbivudine, tenofovir, valacyclovir trifluorothymidine, valganciclovir, zalcitabine (DDC), zidovudine (AZT); f) Non-nucleoside analogues: Amantadine, ateviridine, capvirine, diarylpyrimidines (etravirine, rilpivirine), deraviridine, docosanol, emimivirine, efavirenz, phosphonocarboxylic acid, imiquimod, interferon alpha, cloviramide, lodeadenosine, thiamethoxazole, nevirapine, NOV-205, pegylated interferon alpha, podophyllotoxin, rifampin, amantadine, requimod (R-848), acetamipridamole; g) Protease inhibitors: Ampravir, atazanavir, bosavirin, darunavir, fosanavir, indinavir, lopinavir, nelfinavir, pleconaril, ritonavir, saquinavir, telanavir (VX-950), telanavir; h) Other types of antiviral drugs: anti-enzymes, arbidol, calanolides A. Phytosine, Phytophytic N, Diarylpyrimidine, Epigallocatechin Gallate (EGCG), Sodium Phoscarboxylate, Griffon, Tarivelin (Veramidin), Hydroxyurea, KP-1461, Mitefocin, Precocinone, Synthesis Inhibitor, Ribavirin, Seliciclib; 5) Radioactive isotopes used in radiotherapy. Examples of radioactive isotopes (radionuclides) include 3H, 11C, 14C, 18F, 32P, 35S, 64Cu, 68Ga, 86Y, 99Tc, 111In, 123I, 124I, 125I, 131I, 133Xe, 177Lu, 211At, or 213Bi. Radioactive isotope-labeled antibodies can be used in receptor-targeted imaging experiments or in targeted therapy using antibody-drug conjugates as described in this invention (Wu et al., Nature Biotechnology 2005, 23(9): 1137-1146). Cell-binding molecules, such as antibodies, can bind to ligands and bind, complex, or form complexes with radioactive metals using the methods described in the literature (Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, NY, Pubs. (1991)). Ligands that can complex metal ions include DOTA, DOTP, DOTMA, DTPA, and TETA (Macrocyclics, Dallas, TX), etc. 6) Other cell-binding molecules-drug conjugates as synergists. Preferred cytotoxic molecules on synergistic conjugates include: camptothecin analogs, maytansine alkaloid analogs, paclitaxel (taxane) analogs, CC-1065 analogs, daunorubicin and doxorubicin compounds, amatoxin analogs, benzodiazepines (such as pyrrole benzodiazepines (PBD), tomatine, atrazomycin, indolebenzothiazide diazepam, imidazole benzothiazide diazepam, or oxazolidinyl benzodiazepine dimers), galactocytocin and enediyne antibiotic compounds, actinomycin, diazoserine, bleomycin, epirubicin, tamoxifen, idarubicin, sea haretoxin, and auristatins (such as monomethyl auristatin E, MMAE, MMAF, auristatin PYE, auristatin TP, auristatin 2-AQ, 6-AQ, EB (AEB), and EFP). (AEFP), docalamicin, geldmycin, methotrexate, thiotepa, vincristine, vinblastine, hemiasterlin, nalsalimum, chlortetracycline, radiosumin, spartina alterniflora, microsclerodermin, thiophene cetylamine, esperidone, PNU-159682, and their analogues and derivatives; 7) Other immunotherapeutic agents such as: imiquimod, interferon (e.g., α, β), granulocyte colony-stimulating factor, cytokines, interleukins (IL-1-IL-35), antibodies (e.g., trastuzumab, pertuzumab, bevacizumab, cetuximab, panitumumab, infliximab, adalimumab, balichimab, dalizumab, omalizumab, PD-1 or PD-L1), protein-binding drugs (e.g., abraxane), antibodies conjugated with the following drugs: chachiomycin derivatives, maytansine derivatives (DM1 and DM4), CC-1065, SN-38, ixotecan, topotecan, topoisomerase I inhibitors, docamycin, PBD or IGN minor groove binders, potent paclitaxel derivatives, doxorubicin, aurestatin antimitotic drugs, such as trastuzumab-DM1, trastuzumab deruxtecan(DS-8201a),Inotuzumab ozogamicin,Brentuximab vedotin,Sacituzumab govitecan,Glembatumumab vedotin,Lorvotuzumab mertansine,AN-152 LMB2,TP-38,VB4-845,Cantuzumab mertansine, AVE9633,SAR3419,CAT-8015 (anti-CD22), IMGN388, Mirvetuximab soravtansine (IMGN853), Enfortumab vedotin, Milatuzumab-doxorubicin, SGN-75 (anti-CD70), anti-Her3-ixotecan, anti-Trop2- Ecinotecan, anti-CD79b-MMAE, anti-Her2-MMAE, anti-Trop2-MMAE, anti-Her2-MMAF, anti-Trop2-MMAF, anti-CD22-cazithromycin derivative, anti-CD22-MMAE, anti-Her2-aurestatin derivative, anti-Muc1-aurestatin derivative, anti-cMet-aurestatin derivative or anti-Claudin18.2-aurestatin derivative; 8) Pharmaceutically acceptable salts, acids, or derivatives of any of the above-mentioned drugs.

[0140] In another synergistic immunotherapy, antibodies against immune checkpoint inhibitors, TCR (T cell receptor) T cells, CAR (chimeric antigen receptor) T cells or B cell receptor (BCR) cells, natural killer (NK) cells or cytotoxic cells, or antibodies against -CD3, CD4, CD8, CD16 (FcγRIII), CD19, CD20, CD22, CD25, CD27, CD30, CD33, CD37, CD38, CD40, CD40L, CD45RA, CD45RO, CD56, CD57, CD57 bright, CD70, CD79, CD79b, CD123, CD125, CD138, TNFβ, Fas ligands, MHC class I molecules (HLA-A, B, C), VEGF or NKR-P1 antigens are preferably used in synergistic therapy with conjugates of this patent.

[0141] In another embodiment, a pharmaceutical composition comprising a conjugate of the therapeutically effective dose of the structure of (I) to (VII), or any other conjugate described in this invention, may be administered synergistically with other therapeutic agents, such as chemotherapeutic agents, radiotherapeutic agents, immunotherapeutic agents, autoimmune disease therapeutic agents, anti-infective agents, or other conjugates, to effectively treat or prevent cancer, autoimmune diseases, or infectious diseases. The synergist is preferably selected from one or more of the following groups of drugs: abatacept, abiraterone acetate, acetaminophen / hydrocodone, adalimumab, ADXS31-142, ADXS-HER2, afatinib maleate, alenmab, trastuzumab, amphetamine / dextromethorphan, anastrozole, atazanavir, atezolizumab, atorvastatin, axitinib, beline, BCG Live, bevacizumab, cabazitaxel, cabozantin, bexarotin, blinatumomab, bortezomib, bosutinib, brentuximab Vedotin, Budesonide, Budesonide / Formoterol, Buprenorphine, Capecitabine, Carfilzomib, Celecoxib, Cetuximab, Cyclosporine, Cinacalcet, Crizotinib, Dabigatran, Dabrafenib, Afadabetin, Derrenavir, Imatinib Mesylate, Dasatinib, Denisil Interleukin, Denosema, Sodium Valproate, Dexlansoprazole, Dextromethorphan, Dinutuximab, Doxycycline, Duloxetine, Emtricitabine / Rilpivirine / Tenofovir Disoproxil Fumarate, Emtricitabine / Tenofovir / Efavirenz, Enoxaparin, Enzalutamide, Epoxetine Afa, Erlotinib Esomeprazole, Ezetimibe, Ezetimibe / Simvastatin, Fenofibrate, Filgrastim, Fingolimod, Fluticasone Propionate, Fluticasone / Sameterol, Fulvestrant, Gefitinib, Glatiramer, Goserelin Acetate, Imatinib, Tiimomab, Ibrutinib, Insulin Aspart, Insulin Detemir, Insulin Glargine, Insulin Lispro, Interferon β1a, Interferon β1b, Lapatinib, Ipilimumab, Ipratropium Bromide / Salbutamol, Lanleptide Acetate, Lenalidomide, Combined Benzoylsulfonate, Letrozole, Levothyroxine, Levothyroxine, Lidocaine, Linezolid, LiraglutideMEDI4736, Memantine, Methyl acetate, Metoprolol, Modafinil, Mometasone, Nirotinib, Nivolumab, Olfamumab, Olitazolizumab, Pazopanib, Pembrolizumab, Pemetrexed, Pertuzumab, Pneumococcal conjugate vaccine, Pomalidomide, Pregabalin, Quetiapine, Rabeprazole radium chloride 223, Raloxifene, Ralcifen, Ramirez, Ranibizumab, Regorafenib, Rituximab, Rivaroxaban, Romidixin, Rosuvastatin, Russoteinib phosphate, Salbutamol, Severamine, Sildenafil, Silt uximab, sitagliptin, sitagliptin / metformin, sofenacin, sorafenib, sunitinib, tadalafil, tamoxifen, telaprevir, tesimolimus, tenofovir / emtricitabine, testosterone gel, thalidomide (Talidex), tiotropium bromide, toremifene, trametinib, trastuzumab, tretinoin, eutecticumab, valsartan, vandetanib, vemurafenib, vorinostat, aflibercept, Zostavax and their analogues, derivatives, pharmaceutically acceptable salts, carriers, diluents or excipients, or combinations thereof.

[0142] Furthermore, this invention also relates to a method for preparing the conjugates. The conjugates of this invention can be prepared by methods well known to those skilled in the art. The camptothecin analogue in the conjugate can be prepared by employing or applying the synthetic methods described below, or synthesized with modifications by those skilled in the art. Suitable modifications and substitutions will be obvious and well known to those skilled in the art and can be readily obtained from the scientific literature. These methods can be found in: RC Larock, Comprehensive Organic Transformations, 2nd Edition, Wiley-VCH Publishers, 1999.

[0143] When carrying out the reactions described in this application, it may be necessary to protect some of the active functional groups, such as hydroxyl, amino, imine, thiol, and carboxyl groups, to avoid side reactions. Common methods for protecting functional groups can be found in PG Wuts and TW Greene, Greene's Protective Groups in Organic Synthesis, Wiley-Interscience; 4th edition (2006). Some reactions may require the presence of acids or bases, or in suitable solvents. There are no particular limitations on acids, bases, and solvents; any known acids, bases, and solvents can be used here, provided there are no adverse effects. Reactions can be carried out over a wide temperature range. Generally, reaction temperatures between -80°C and 150°C are easier to operate at (between room temperature and 100°C is better). The reaction time can also vary considerably, depending on various factors, especially the reaction temperature and the nature of the solvent used. Generally, under ideal conditions, a reaction time of 3 to 20 hours is sufficient.

[0144] Post-reaction processing can be performed using conventional methods. For example, the reaction product can be recovered by evaporating the solvent from the reaction system. Alternatively, if necessary, after evaporating the solvent, the residue can be poured into water, extracted with a water-immiscible organic solvent, and finally, the extraction solvent can be evaporated. Furthermore, if needed, the product can be further purified using various common methods, such as recrystallization, precipitation, or various chromatographic methods, such as column chromatography and preparative thin-plate chromatography. The synthesis of the camptothecin analogue and its conjugates in this invention is shown in Figures 1-32.

[0145] The conjugation of the cell-binding molecule of the present invention with camptothecin analogues is further described in the embodiments. Of course, the present invention is not limited to the following embodiments.

[0146] experiment []

[0147] The following examples further describe the invention and are not intended to limit the scope of the invention. Unless otherwise stated, the cell lines described in the following examples were cultured according to the conditions specified by the American Type Culture Collection (ATCC), Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DMSZ) in Braunswehr, Germany, or the Shanghai Institute of Cell Culture, Chinese Academy of Sciences. Unless otherwise stated, cell culture reagents were from Invitrogen Corp. Amino acids and their derivatives, as well as pre-packaged resins, were from Merck Chemical International, Synthetech, Peptide International, Chembridge International, or Sigma-Aldrich (Merck). Some linkers include NHS ester / maleimide linkers (AMAS, BMPS, GMBS, MBS, SMCC, EMCS or Sulfo-EMCS, SMPB, SMPH, LC-SMCC, Sulfo-KMUS, SM(PEG)4, SM(PEG)6, SM(PEG)8, SM(PEG)12, SM(PEG)24); NHS ester / pyridyl dithiol (SPDP, LC-SPDP or Sulfo-LC-SPDP, SMPT, Sulfo-LC-SMPT); NHS ester / haloacetyl (SIA, SBAP, SIAB or Sulfo-SIAB). NHS ester / diazo (SDA or Sulfo-SDA, LC-SDA or Sulfo-LC-SDA, SDAD or Sulfo-SDAD); maleimide / acehydrazine (BMPH, EMCH, MPBH, KMUH); pyridyl dithiol / acehydrazine (PDPH); isocyanate / maleimide (PMPI) were purchased from Thermo Fisher Scientific Co. SPDB and SPP linkers were prepared according to the reference (Cumber, A. et al., Bioconjugate Chem., 1992, 3, 397-401). Roche's trastuzumab was purchased from a Chinese pharmacy. The Trop-2 antibody was a bioderive of Sacituzumab, manufactured in-house. The EGFR antibody, Nimotuzumab, was purchased from a Chinese pharmacy. PEG and PEG derivatives were purchased from Biomatrik Inc., Jiaxing, Zhejiang Province, China. Topotecan and its derivatives or key intermediates were sourced from several companies, including Chengdu Tianyuan Natural Products Co., Ltd. and Suzhou Guangming Biomedical Co., Ltd., both in China. Laboratory animals were purchased from the National Mouse Model Resource Center through GemPharmatech Co., Ltd. in Nanjing, China, and SLAC Laboratory Animal Co., Ltd. in Shanghai, China. T-DM1 was purchased from Roche through a pharmacy in Hong Kong, China.All other reagents and solvents purchased were of the highest available grade and could be used without further purification. EDC (EDCI), PFP, HATU, TATU, PyBrOP, DIPEA, TEA, PPTS, DMAP, BrOP, p-TSA, DTT, EDTA, TCEP, NHS, trifluoroacetic acid, Ellman's reagent, Traut's reagent (2-iminothiocyclopentane), γ-thiobutyrolactone, and all other chemical reagents and anhydrous solvents were from Sigma-Aldrich International (Merck) or Aladdin Chemical (Shanghai) Ltd. All anhydrous solvents were obtained through commercial routes and stored in Sure-seal bottles under nitrogen. Preparative HPLC separations (acetonitrile / water, containing formic acid or trifluoroacetic acid) were performed on a Varaiin PreStar HPLC system. NMR spectra were obtained on a Bruker 500MHz instrument. Chemical shifts (δ) are reported in parts per million (ppm), with tetramethylsilane (0.00 ppm) as a reference. Coupling constants (J) are reported in Hz. Mass spectrometry data were obtained on a Waters Xevo G2 QTOF mass spectrometer equipped with an Acquity UPLC separation module and an Acquity TUV detector.

[0148] Example 1. Synthesis of (S)-(1-((4-ethyl-4,9-dihydroxy-3,14-disideoxy-3,4,12,14-tetrahydro-1H-guanan[3', 4';6,7]indolo[[1,2-b]quinoline-10-yl)methyl)hexahydropyridin-4-yl)aminocarbamate tributyl ester (2). A solution of 4-tert-butoxycarbonylaminohexahydropyridine (4.4 g, 21.9 mmol) and 37% formaldehyde (1.8 g, 21.9 mmol) in acetic acid (10 mL) was added to a solution of 10-hydroxycamptothecin (2 g, 5.49 mmol). The reaction mixture was heated to approximately 60 °C and stirred for 2 hours, after which the acetic acid was removed. Recrystallization from 10 mL of methanol gave compound 2 (2.1 g, 68% yield) as a yellow powder. ESI-MS m / z: [M + H] + C 31H 36N 4O 7: Calculated 577.26; Found 577.26.

[0149] Example 2. Synthesis of (S)-10-((4-aminohexahydropyridin-1-yl)methyl)-4-ethyl-4,9-dihydroxy-1H-guanano[3',4';6,7]indolano[1,2]-b]quinoline-3,14(4H,12H)-dione (3). Compound 2 (150 mg, 0.26 mmol) was dissolved in a mixture of dichloromethane and trifluoroacetic acid (2 mL / 6 mL) and stirred at room temperature for 1 hour. The mixture was then concentrated and dried under vacuum to give compound 3 (120 mg, 100% yield) as a yellow solid. ESI-MS m / z: [M + H] + C 26H 28N 4O 5: Calculated 477.21; Found 477.21.

[0150] Example 3. Synthesis of compound (S)-4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)-N-(1-((4-ethyl-4,9-dihydroxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-10-yl)methyl)hexahydropyridin-4-yl)butyramine (5). Compound 3 (62 mg, 0.13 mmol) and pentafluorophenyl 4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyrate (compound 4, 56 mg, 0.16 mmol) were dissolved in DMF (5 mL), cooled to approximately 0 °C, and then N,N-diisopropylethylamine (45 μL, 0.16 mmol) was added. The reaction was heated to room temperature, stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 5 (44 mg, 54% yield). ESI-MS m / z: [M + H] +C 34H 35N 5O 8: Calculated value 642.25; Found value 642.25.

[0151] Example 4. Synthesis of (S)-pentafluorophenyl 30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-27,31-dioxy-2,5,8,11,14,17,20,23-octaoxy-26,32-diazahexadecane-36-carboxylate (7). Compound 6 (100 mg, 0.13 mmol) was dissolved in a solution of dichloromethane (5 mL), and pentafluorophenol (48 mg, 0.26 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (50 mg, 0.26 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours, diluted with dichloromethane (50 mL), washed with water (2 × 10 mL), dried over sodium sulfate, filtered, and concentrated to give compound 7 (120 mg, 100% yield). ESI-MS m / z: [M + H] +C 40H 57F 5N 4O 15: Calculated value 929.37; Found value 929.37.

[0152] Example 5. Synthesis of (S)-2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-N1-(4-((1-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4';6,7]indolano[1,2-b]quinoline-10-yl)methyl)hexahydropyridin-4-yl)amino)-4-dioxybutyl)-N5-(2,5,8,11,14,17,20,23-octa-dioxytetradecane-25-yl)pentanediamine (8). Compound 3 (60 mg, 0.126 mmol) and compound 7 (97 mg, 0.105 mmol) were dissolved in DMF (5 mL) and cooled to approximately 0 °C. Then, N,N-diisopropylethylamine (37 μL, 0.21 mmol) was added. The reaction was heated to room temperature and stirred for 2 hours. The mixture was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 8 (50 mg, 39% yield). ESI-MS m / z: [M + H] +C 60H 84N 8O 19: Calculated value 1221.59; Found value 1221.59.

[0153] Example 6. Synthesis of 3-(2-(2-(2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)ethoxy)ethoxy)propionate tert-butyl ester (10). Compound 9 (1.32 g, 5.7 mmol) was dissolved in DMF (10 mL), and 4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyric acid (1.04 g, 5.7 mmol) was added, followed by HATU (2.6 g, 6.8 mmol) and triethylamine (0.96 mL, 6.8 mmol) in sequence. The mixture was stirred at room temperature for 1 hour, diluted with dichloromethane (100 mL), washed with water (2 × 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (ethyl acetate / petroleum ether) to give compound 10 (1.8 g, 80% yield). ESI-MS m / z: [M + H] + C 19H 30N 2O 7: Calculated value 399.21; Found value 399.21.

[0154] Example 7.3 Synthesis of (2-(2-(2-(4-(2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)butyramine)ethoxy)ethoxy)propionic acid (11). Compound 10 (0.40 g, 1.0 mmol) was dissolved in a mixed solution of dichloromethane and trifluoroacetic acid (12 mL / 4 mL) and stirred at room temperature for 1 hour. The mixture was then concentrated twice with dichloromethane and dried under vacuum to give compound 11 (0.34 g, 100% yield) as a yellow solid. ESI-MS m / z: [M + H] + C 15H 22N 2O 7: Calculated value 343.14; Found value 343.14.

[0155] Example 8. Synthesis of pentafluorophenyl 3-(2-(2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butimino)ethoxy)ethoxy)propionate (12). Compound 11 (0.34 g, 1.0 mmol) was dissolved in a solution of dichloromethane (30 mL), and pentafluorophenol (0.46 g, 2.5 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.57 g, 3.0 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours, diluted with dichloromethane (50 mL), washed with water (200 mL), dried over sodium sulfate, filtered, and concentrated to give compound 12 (0.51 g, 100% yield). ESI-MS m / z: [M + H] +C 21H 22F 5N 2O 7: Calculated value 509.13; Found value 509.13.

[0156] Example 9. Synthesis of N,N-dimethylhexahydropyridine-4-amine (13). N-Boc hexahydropyridone (10 g, 0.05 mol) was dissolved in methanol (100 mL), and dimethylamine aqueous solution (25 mL, 0.22 mol) was added. Catalytic hydrogenation with 10% Pd / C (1 g) was carried out overnight at room temperature with stirring. After filtration, the filtrate was concentrated and co-concentrated three times with dichloromethane (3 × 80 mL), and dried on a vacuum pump to remove all dimethylamine. Hydrochloric acid / methanol (4 M, 50 mL) was added to the residue, and the mixture was stirred at room temperature for 30 minutes. A large amount of white solid precipitated. The mixture was filtered to give white solid 13 (9 g, 90% yield). ESI-MS m / z: [M + H] + C 7H 16N 2, calculated value 129.13; measured value 129.13.

[0157] Example 10. Synthesis of (9H-fluorene-9-yl)methyl 4-(dimethylamino)hexahydropyridine-1-carboxylate (14). Compound 13 (2.0 g, 9.9 mmol) was dissolved in a mixture of 1,4-dioxane and water (30 mL / 50 mL), and sodium bicarbonate (2.5 g, 29.8 mmol) was added. The mixture was cooled to 0 °C, and a solution of 9-pyruvylmethoxycarbonyl chloride (3.1 g, 11.9 mmol) in 1,4-dioxane (10 mL) was added dropwise. After the addition was complete, the temperature was gradually raised to room temperature, and the reaction mixture was stirred for 1 hour. 100 mL of 1 M hydrochloric acid was added, and the mixture was washed with ethyl acetate (3 × 50 mL). The aqueous phase was adjusted to pH ~10 with sodium carbonate, and then extracted with dichloromethane (3 × 50 mL). The combined organic phases were washed with water (50 mL), dried over sodium sulfate, filtered, concentrated, and purified by column chromatography (methanol / dichloromethane) to give compound 14 (2.75 g, 79% yield). ESI-MS m / z: [M + H] +C 22H 26N 2O 2, calculated value 351.20; measured value 351.20.

[0158] Example 11. Synthesis of (S)-tertiary butyl (1-((4-(hydroxymethyl)phenyl)amino)-1-sideoxypropyl-2-yl)aminocarbamate (15). p-Aminobenzyl alcohol (5.0 g, 0.04 mol) and Boc-L-alanine (8.0 g, 0.042 mol) were dissolved in anhydrous THF (100 mL), and 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (11 g, 0.044 mol) was added and stirred overnight at room temperature. The reaction mixture was poured into water (300 mL), extracted with ethyl acetate (3 × 100 mL), the combined organic phases were washed with water (100 mL), dried over sodium sulfate, filtered, and concentrated. The crude product was wet-milled with ethyl acetate / petroleum ether (1:3), filtered, and compound 15 (9.8 g, 84% yield) was given as a white solid. ESI-MS m / z: [M + H] + C 15H 22N 2O 4: Calculated value 295.16; Found value 295.16.

[0159] Example 12. Synthesis of (S)-tertiary butyl (1-((4-(bromomethyl)phenyl)amino)-1-sideoxypropyl-2-yl)aminocarbamate (16). Compound 3 (3.5 g, 11.9 mmol) and carbon tetrabromide (5.9 g, 17.8 mmol) were dissolved in dichloromethane (80 mL), cooled to approximately 0 °C, and triphenylphosphine (4.7 g, 17.8 mmol) was added. The reaction mixture was heated to room temperature and stirred for 30 minutes, then 20 g of silica gel was added, mixed, dried on a rotary condenser, and loaded onto a silica gel column (100 g silica gel). The mixture was then dissolved in petroleum ether / ethyl acetate to give compound 16 (2.6 g, 62% yield). ESI-MS m / z: [M + H] + C15H21BrN2O3: Calculated value 357.07; Found value 357.07.

[0160] Example 13. Synthesis of (S)-1-(((9H-fluorene-9-yl)methoxy)carbonyl)-N-(4-(2-(tert-butoxycarbonyl)amino)propionic acid)benzyl)-N,N-dimethylhexahydropyridine-4-bromoamine (17). Compound 16 (2.3 g, 6.4 mmol) and compound 14 (2.7 g, 7.7 mmol) were dissolved in anhydrous THF (50 mL) and stirred overnight at room temperature. After removing most of the THF using a rotary condenser, ethyl acetate (50 mL) was added to the residue. The resulting slurry was filtered to give a white solid (4.5 g, 100% yield). ESI-MS m / z: M + C 37H 47N 4O 5: Calculated 627.35; Found 627.35.

[0161] Example 14. Synthesis of (S)-N-(4-(2-(tert-butoxycarbonyl)amino)propionic)benzyl)-N,N-dimethylhexahydropyridine-4-bromoamine (18). Compound 17 (1.0 g, 1.41 mmol) was dissolved in 5 mL of DMF, and hexahydropyridine (1 mL) was added. After stirring at room temperature for 30 minutes, 30 mL of ethyl acetate was added, and the mixture was stirred for 10 minutes. The mixture was filtered to give a white powdery solid (550 mg, 80% yield). ESI-MS m / z: M + C 22H 37N 4O 3: Calculated value 405.29; Found value 405.29.

[0162] Example 15. Synthesis of N-(4-((S)-2-((tert-butoxycarbonyl)amino)propionic acid)benzyl)-1-((S)-4-ethyl-4,9-dihydroxy-3,14-di-side-oxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-10-yl)methyl)-N,N-dimethylhexahydropyridine-4-bromoamine (19). Compound 18 (550 mg, 1.13 mmol) and a 37% formaldehyde (92 mg, 1.13 mmol) acetic acid solution (5 mL) were added to a solution of 10-hydroxycamptothecin (375 mg, 1.03 mmol) in acetic acid (5 mL). The mixture was heated to approximately 65 °C and stirred for 1 hour, then concentrated with anhydrous methanol. Recrystallization with dichloromethane and a small amount of methanol gave compound 19 (0.5 g, 63% yield) as a yellow powder. ESI-MS m / z: M + C 43H 53N 6O 8: Calculated 781.39; Found 781.39.

[0163] Example 16. Synthesis of N-(4-((S)-2-aminopropylamino)benzyl)-1-(((S)-4-ethyl-4,9-dihydroxy-3,14-disideoxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-10-yl)methyl)-N,N-dimethylhexahydropyridine-4-bromoamine (20). Compound 19 (50 mg, 0.058 mmol) was dissolved in a mixed solution of dichloromethane and trifluoroacetic acid (2 mL / 6 mL) and stirred at room temperature for 30 minutes. The mixture was then concentrated and dried under vacuum to give compound 20 (44 mg, 100% yield) as a yellow solid. ESI-MS m / z: M + C 38H 45N 6O 6: Calculated value 681.34; Found value 681.34.

[0164] Example 17. Synthesis of N-(4-((S)-2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)propamido)benzyl)-1-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-10-yl)methyl)-N,N-dimethylhexahydropyridine-4-carboxylic acid ammonium (21). Compound 20 (88 mg, 0.116 mmol) and compound 4 (49 mg, 0.140 mmol) were dissolved in DMF (5 mL), cooled to approximately 0 °C, and then dissolved in N,N-diisopropylethylamine (40 μL, 0.232 mmol). The reaction was heated to room temperature. After stirring for 2 hours, the mixture was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 21 (66 mg, 68% yield). ESI-MS m / z: M +C 46H 52N 7O 9: Calculated value 846.38; Found value 846.38.

[0165] Example 18. Synthesis of N-(4-((9S,17S)-9-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-17-methyl-6,10,15-trioxy-2-oxa-5,11,16-triazaoctadecylamine)benzyl)-1-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-10-yl)methyl)-N,N-dimethylhexahydropyridine-4-carboxylic acid ammonium (22). Compound 20 (44 mg, 0.058 mmol) and compound 7 (60 mg, 0.065 mmol) were dissolved in DMF (5 mL), cooled to approximately 0 °C, and then dissolved in N,N-diisopropylethylamine (20 μL, 0.116 mmol). The reaction was heated to room temperature. After stirring for 2 hours, the mixture was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 22 (51 mg, 58% yield). ESI-MS m / z: M +C 72H 101N 10O 20: Calculated 1425.72; Found 1425.72.

[0166] Example 19.1 Synthesis of (2-amino-4-fluoro-5-methoxyphenyl)-2-chloroethyl ketone (24d). A solution of 3-fluoro-4-methoxyaniline (5 g, 35.4 mmol) dissolved in dichloromethane (20 mL) was added dropwise to a solution of boron trichloride (1 M dichloromethane, 38.9 mL) cooled in ice water. The reaction was stirred for 10 minutes, and then chloroacetonitrile (3.2 g, 42.5 mmol) and aluminum trichloride (5.2 g, 38.9 mmol) were added. After the addition was complete, the reaction was heated to room temperature and then refluxed overnight. The reaction mixture was then cooled to approximately 0 °C, quenched with 2 M hydrochloric acid (80 mL), and stirred continuously at room temperature for 2 hours. The layers were separated, and the aqueous phase was extracted with dichloromethane (3 × 80 mL). The combined organic phases were washed with water (100 mL), dried over sodium sulfate, filtered, concentrated, purified on a silica gel column, and dissolved in petroleum ether / ethyl acetate to give compound 24d (2 g, 26% yield) as a yellow solid. ESI-MS m / z: [M + H] +C 9H 9ClFNO 2: Calculated value 218.03; Measured value 218.03.

[0167] Example 20. Synthesis of (S)-11-(chloromethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (26d). Compound 24d (0.50 g, 2.29 mmol) and compound 25 (0.57 g, 2.19 mmol) were dissolved in anhydrous toluene (40 mL), and p-toluenesulfonic acid (42 mg, 0.219 mmol) was added. The suspension was heated under reflux for 2 days and then cooled to room temperature. After removing approximately two-thirds of the toluene, the residue was filtered, and the filter cake was washed with dichloromethane and air-dried to give compound 26d (0.7 g, 72% yield) as a gray powder. ESI-MS m / z: [M + H] + C 22H 18ClFN 2O 5: Calculated 445.09; Found 445.09.

[0168] Example 21. Synthesis of N-(4-((S)-2-((tert-butoxycarbonyl)amino)propionic)benzyl)-1-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-chloroamine (27). A mixture of compound 26d (218 mg, 0.49 mmol) and compound 18 (200 mg, 0.49 mmol) in DMF (5 mL) was stirred for 30 minutes at 0 °C, followed by the addition of triethylamine (63 μL, 0.45 mmol) and stirring for another hour. The reactants were concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 10 (240 mg, 59% yield) as a yellow solid. ESI-MS m / z: M +C 44H 54FN 6O 8: Calculated value 813.40; Found value 813.40.

[0169] Example 22. Synthesis of N-(4-((S)-2-aminopropylamino)benzyl)-1-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-aminoonium (28). Compound 27 (50 mg, 0.06 mmol) was dissolved in a mixed solution of dichloromethane and trifluoroacetic acid (2 mL / 6 mL) and stirred at room temperature for 30 minutes. The mixture was then concentrated and dried under vacuum to give compound 28 (42 mg, 100% yield) as a yellow solid. ESI-MS m / z: M +C 39H 46FN 6O 6: Calculated value 713.35; Found value 713.35.

[0170] Example 23. Synthesis of N-(4-((30S,38S)-30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-38-methyl-27,31,36-trioxy-2,5,8,11,14,17,20,23-octaoxa-26,32,37-triazanonadecanyl)benzyl)-1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-carboxylic acid ammonium (29). Compound 28 (47 mg, 0.060 mmol) and compound 7 (60 mg, 0.066 mmol) were dissolved in DMF (5 mL), cooled to approximately 0 °C, and then dissolved in N,N-diisopropylethylamine (21 μL, 0.12 mmol). The reaction was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 29 (23 mg, 25% yield). ESI-MS m / z: M +C 73H 102FN 10O 20: Calculated value 1457.73; Found value 1457.73.

[0171] Example 24. Synthesis of (S)-11-(aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (30). Compound 26d (80 mg, 0.18 mmol) was dissolved in ethanol (5 mL), hexamethylenetetramine (76 mg, 0.54 mmol) was added, and the mixture was refluxed for 90 minutes and then cooled to room temperature. Concentrated hydrochloric acid (100 μL) was added, and the mixture was stirred for 30 minutes. After concentration, a grayish-white solid was obtained, which was purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 30 (40 mg, 52% yield). ESI-MS m / z: [M + H] + C 22H 20FN 3O 5: Calculated value 426.14; Found value 426.14.

[0172] Example 25. Synthesis of (S)-2-(4-(2,5-di-side-oxy-2,5-dihydro-1H-pyrrole-1-yl)butyramine)-N1-(4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,4,12,14-tetrahydro-1H-guanan-[3',4':6,7]indolo[1,2-b]quinoline-10-yl)methyl)amino)-4-side-oxybutyl)-N5-(2,5,8,11,14,17,20,23-octaoxapentanodecane-25-yl)pentanediamine (31). Compound 30 (40 mg, 0.094 mmol) and compound 7 (120 mg, 0.13 mmol) were dissolved in DMF (5 mL) and cooled to approximately 0 °C. Then, N,N-diisopropylethylamine (33 μL, 0.188 mmol) was added. The reaction was heated to room temperature. After stirring for 2 hours, the mixture was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 31 (55 mg, 50% yield). ESI-MS m / z: [M + H] +C 56H 76FN 7O 19: Calculated value 1170.52; Found value 1170.52.

[0173] Example 26. Synthesis of third butyl (1-methylhexahydropyridin-4-yl)carbamate (32). 4-(tert-butoxycarbonylamino)hexahydropyridine (2 g, 10 mmol) was dissolved in methanol (30 mL), then 37% formaldehyde (1.6 g, 20 mmol) was added, and catalytic hydrogenation (1 atm) with 10% Pd / C (0.2 g) was carried out overnight, stirred, and filtered. The filtrate was concentrated to give compound 32 (2.1 g, 100% yield). ESI-MS m / z: [M + H] +C 11H 22N 2O 2: Calculated value 215.17; Found value 215.17.

[0174] Example 27. Synthesis of (S)-4-((tert-butoxycarbonyl)amino)-1-((4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-di-side-oxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylhexahydropyridine-1-onium chloride (33). A DMF (3 mL) solution of compound 26d (50 mg, 0.112 mmol) and compound 32 (26 mg, 0.123 mmol) was stirred at room temperature for 2 hours. The reaction solution was purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 6 (33 mg, 47% yield). ESI-MS m / z: [M] +C 33H 40FN 4O 7: Calculated value 623.29; Found value 623.29.

[0175] Example 28. Synthesis of (S)-4-amino-1-((4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylhexahydropyridine-1-onium (34). Compound 33 (30 mg, 0.053 mmol) was dissolved in a mixed solution of dichloromethane and trifluoroacetic acid (3 mL / 1 mL) and stirred at room temperature for 30 minutes. The mixture was then concentrated and dried under vacuum to give compound 34 (33 mg, 100% yield). ESI-MS m / z: [M] +C 28H 32N 4O 5: Calculated 477.21; Found 477.21.

[0176] Example 29.4 Synthesis of ((S)-30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-27,31-dioxy-2,5,8,11,14,17,20,23-octaoxa-26,32-diazahexadecanoylamine)-1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylhexahydropyridine-1-carboxylic acid onium (35). Compound 34 (30 mg, 0.053 mmol) and compound 7 (60 mg, 0.079 mmol) were dissolved in DMF (5 mL) and cooled to approximately 0 °C. Then, N,N-diisopropylethylamine (18 L, 0.106 mmol) was added. The reaction was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 35 (15 mg, 21% yield). ESI-MS m / z: [M] +C 62H 88FN 8O 19: Calculated value 1267.61; Found value 1267.61.

[0177] Example 30. Synthesis of methyl (9H-fluorene-9-yl)4-methylpiperazine-1-carboxylic acid (36). 1-Methylpiperazine (5.0 g, 50.0 mmol) was dissolved in a mixture of 1,4-dioxane and water (60 mL / 100 mL), sodium bicarbonate (12.6 g, 150 mmol) was added, and the mixture was cooled to 0 °C. Then, a solution of 9-piperazine methoxycarbonyl chloride (15.5 g, 60.0 mmol) in 1,4-dioxane (20 mL) was added dropwise. After the addition was complete, the temperature was gradually increased to room temperature, and the reaction mixture was stirred for 3 hours. 300 mL of 1M hydrochloric acid was added, and the mixture was washed with ethyl acetate (2 × 100 mL). The aqueous phase was adjusted to pH ~10 with sodium carbonate, and then extracted with ethyl acetate (2 × 100 mL). The combined organic phases were washed with water (250 mL), dried over sodium sulfate, filtered, concentrated, and purified by column chromatography (methanol / dichloromethane) to give compound 2 (6.5 g, 40% yield). ESI-MS m / z: [M + H] +C 20H 22N 2O 2, calculated value 323.17; measured value 323.19.

[0178] Example 31. Synthesis of (S)-4-(((9H-fluorene-9-yl)methoxy)carbonyl)-1-(4-(2-(tert-butoxycarbonyl)amino)propionic acid)benzyl)-1-methylpiperazine-1-bromium (37). Compound 16 (2.3 g, 6.4 mmol) and compound 36 (2.1 g, 6.4 mmol) were dissolved in anhydrous THF (100 mL) and stirred overnight at room temperature. After removing most of the THF using a rotary condenser, ethyl acetate (200 mL) was added to the residue. The resulting slurry was filtered to give a white solid (3.8 g, 87% yield). ESI-MS m / z: M + C 35H 43N 4O 5: Calculated 599.32; Found 599.32.

[0179] Example 32. Synthesis of (S)-1-(4-(2-(tert-butoxycarbonyl)amino)propionic acid)benzyl)-1-methylpiperazine-1-bromodium (38). Compound 37 (3.12 g, 4.6 mmol) was dissolved in 25 mL of DMF, and hexahydropyridine (3 mL) was added. After stirring at room temperature for 2 hours, 200 mL of ethyl acetate was added, and the mixture was stirred for 10 minutes. The mixture was filtered to give a white solid (1.54 g, 77% yield). ESI-MS m / z: M + C 20H 33N 4O 3: Calculated value 377.26; Found value 377.26.

[0180] Example 33.1 Synthesis of 4-((S)-2-((tert-butoxycarbonyl)amino)propionic acid)benzyl)-4-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-di-side-oxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylguanazin-1-onium (39). At 0 °C, a mixture of compound 38 (0.30 g, 0.66 mmol) and compound 26d (0.25 g, 0.56 mmol) in DMF (10 mL) was stirred for 30 min. Then, N,N-diisopropylethylamine (49 μL, 0.28 mmol) was added, and the reaction mixture was heated to room temperature and stirred overnight. The mixture was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 39 (0.40 g, 80% yield). ESI-MS m / z: M +C 42H 50FN 6O 8: Calculated value 785.37; Found value 785.37.

[0181] Example 34.1 Synthesis of 4-(((S)-2-aminopropylamino)benzyl)-4-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-di-side-oxy-3,4,12,14-tetrahydro-1H-guanano[3',4';6,7]indolano[[1,2-b]quinoline-11-yl)methyl)-1-methylguanazin-1-onium (40). Compound 39 (0.30 g, 0.35 mmol) was dissolved in a mixed solution of dichloromethane and trifluoroacetic acid (3 mL / 3 mL) and stirred at room temperature for 30 minutes. The mixture was then concentrated and dried under vacuum to give compound 40 (0.27 g, 100% yield) as a yellow solid. ESI-MS m / z: M + C 37H 42FN 6O 6: Calculated value 477.21; Found value 477.21.

[0182] Example 35.1 Synthesis of 4-((S)-2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)propionamine)benzyl)-4-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylpiperazine (41). Compound 40 (50 mg, 0.065 mmol) and compound 4 (30 mg, 0.098 mmol) were dissolved in DMF (3 mL), and then N,N-diisopropylethylamine (45 μL, 0.26 mmol) was added. The reaction was stirred at room temperature for 30 min, concentrated, and purified by preparative C-18 HPLC (acetonitrile / water containing formic acid) to give compound 41 (37 mg, 61% yield). ESI-MS m / z: M +C 45H 49FN 7O 9: Calculated value 850.36; Found value 850.36.

[0183] Example 36.1-(4-((30S,38S)-30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-38-methyl-27,31,36-trioxy-2,5,8,11,14,17,20,23-octaoxa-26,32,37-triaza-nonadecanyl)benzyl)-4-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylguanazin-1-carboxylic acid onium (42) Compound 40 (70 mg, 0.092 mmol) was dissolved in DMF (2 mL), and a DMF (2 mL) solution of compound 7 (70 mg, 0.092 mmol) was added. Then, HATU (52 mg, 0.138 mmol) and triethylamine (52 µL, 0.368 mmol) were added sequentially, followed by stirring at room temperature for 30 minutes. After concentration, the residue was purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 43 (50.9 mg, 37% yield). ESI-MS m / z: [M+] +C 71H 98FN 10O 20: Calculated value 1429.69; Found value 1429.69.

[0184] Example 37.1 Synthesis of 4-((S)-17-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)-2-methyl-4,14-dioxy-7,10-dioxy-3,13-diazadecylamine)benzyl)-4-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylguanazin-1-carboxylic acid ontium (43). A 1 mL solution of DMF containing compound 40 (0.10 g, 0.13 mmol) and a 2 mL solution of DMF containing compound 12 (66 mg, 0.13 mmol) were mixed, and then N,N-diisopropylethylamine (90 μL, 0.52 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 43 (50.9 mg, 39% yield). ESI-MS m / z: M +C 52H 62N 8O 12: Calculated 1009.45; Found 1009.45.

[0185] Example 38. Synthesis of (S)-3-(tert-butoxycarbonyl)amino)-2-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)propionic acid (44). Methyl dihydro-1H-pyrrole-1-carboxylate (1.52 g, 9.80 mmol) was added to a saturated sodium bicarbonate solution (20 mL) of 2-amino-3-((tert-butoxycarbonyl)amino)propionic acid (1 g, 4.90 mmol) (20 mL) in an ice-water bath. The reaction mixture was stirred for 30 minutes, then poured into a separatory funnel containing 100 mL of ethyl acetate. The organic phase was separated, washed with 50 mL of water and 50 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 4 (1.39 g, 72% yield).

[0186] Example 39. Synthesis of (S)-pentafluorophenyl 3-(tert-butoxycarbonyl)amino)-2-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)propionate (45). To a solution of compound 44 (0.10 g, 0.35 mmol) in dichloromethane (30 mL), pentafluorophenol (97 mg, 0.52 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.13 g, 0.7 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours, diluted with dichloromethane (50 mL), washed with water (200 mL), dried over sodium sulfate, filtered, and concentrated to give compound 45 (0.16 g, 100% yield). ESI-MS m / z: [M + H] +C 18H 15F 5N 2O 6: Calculated value 451.09; Found value 451.09.

[0187] Example 40. Synthesis of compound 46. Compound 40 (0.05 g, 0.065 mmol) and compound 45 (45 mg, 0.10 mmol) were dissolved in DMF (3 mL), and then N,N-diisopropylethylamine (45 μL, 0.26 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 46 (35 mg, 52% yield). ESI-MS m / z: M +C 49H 56FN 8O 11: Calculated value 951.41; Found value 951.41.

[0188] Example 41.1 Synthesis of 4-((S)-2-((S)-3-amino-2-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)propionic)propionic)benzyl)-4-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-1-methylguanazin-1-onium (47). Compound 46 (35 mg, 0.03 mmol) was dissolved in dichloromethane (2 mL) and reacted with trifluoroacetic acid (1 mL). After stirring at room temperature for 1 hour, the reaction mixture was concentrated twice with dichloromethane and dried under vacuum to give compound 47 (30.4 mg, 96% yield). ESI-MS m / z: M + C 44H 48FN 8O 9: Calculated value 851.35; Found value 851.35.

[0189] Example 42. Synthesis of (S)-tertiary butyl (1-(4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-di-side-oxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)hexahydropyridin-4-yl)carbamate (48). Compound 26d (50 mg, 0.11 mmol) was dissolved in DMF (3 mL), and then tert-butyl hexahydropyridine-4-ylaminocarbamate (25 mg, 0.12 mmol) was added. The mixture was stirred at room temperature for 5 hours. The mixture was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 48 (30 mg, 45% yield). ESI-MS m / z: [M + H] + C 32H 37FN 4O 7: Calculated value 609.26; Found value 609.26.

[0190] Example 43. Synthesis of (S)-11-((4-aminohexahydropyridin-1-yl)methyl)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-1H-guanan[3',4':6,7]indolo[1,2-b]quinoline-3,14(4H,12H)-dione (49). Compound 48 (30 mg, 0.03 mmol) was dissolved in dichloromethane (2 mL) and reacted with trifluoroacetic acid (2 mL). After stirring at room temperature for 1 hour, the mixture was concentrated twice with dichloromethane and dried under vacuum to give compound 49 (25.4 mg, 100% yield). ESI-MS m / z: [M + H] + C 27H 30FN 45: Calculated 509.21; Found 509.21.

[0191] Example 44. Synthesis of (S)-2-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-N1-(4-((1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)hexahydropyridin-4-yl)amino)-4-dioxybutyl)-N5-(2,5,8,11,14,17,20,23-octaoxapentano-25-yl)pentadiamine (50). Compound 49 (25.4 mg, 0.05 mmol) was dissolved in DMF (2 mL), compound 7 (38.1 mg, 0.05 mmol) was added, followed by HATU (28.5 mg, 0.08 mmol) and triethylamine (14 µL, 0.1 mmol). The reaction mixture was stirred at room temperature for 1 hour, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 50 (14.4 mg, 23% yield). ESI-MS m / z: [M + H] + C 61H 85FN 8O 19: Calculated value 1253.59; Found value 1253.59.

[0192] Example 45. Synthesis of third butyl bis(2-(2,2,2-trifluoroacetamido)ethyl)aminocarbamate (51). At 0 °C, a solution of ethyl trifluoroacetate (18.75 g, 132 mmol) in dichloromethane (120 mL) was added dropwise over 30 minutes to a solution of diethylenetriamine (6.18 g, 60 mmol) in dichloromethane (60 mL), and then the mixture was heated to room temperature and stirred for 1 hour. A solution of dibutyl dicarbonate (28.78 g, 132 mmol) and triethylamine (13.33 g, 132 mmol) in dichloromethane (60 mL) was added dropwise at room temperature and stirred overnight. The reaction mixture was washed with saturated sodium carbonate (2 × 200 mL), water (2 × 200 mL), and brine (200 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate) to give a white solid (17.4 g, 73.3% yield). ESI-MS m / z: [M + H] +C 13H 19F 6N 3O 4: Calculated value 396.30; Measured value 396.28.

[0193] Example 46. Synthesis of third butyl bis(2-aminoethyl)aminocarbamate (52). Compound 51 (4.28 g, 10.8 mmol) was dissolved in methanol (50 mL) and stirred for 3 hours at room temperature with a solution of sodium hydroxide (5.42 g, 135 mmol) in water (50 mL). The reaction was concentrated, extracted with dichloromethane (3 × 100 mL), the organic phase was washed with brine (100 mL), dried over sodium sulfate, filtered, and concentrated to give compound 3 (1.8 g, 82% yield). ESI-MS m / z: [M + H] + C 9H 21N 3O 2 Calculated value 204.28; Found value 204.12. Example 47.4 Synthesis of 4'-((((tert-butoxycarbonyl)azadiyl)bis(ethane-2,1-diyl))bis(azadiyl))bis(4-sideoxybutyric acid)(53). Compound 52 (1.8 g, 8.8 mmol) was dissolved in dichloromethane (150 mL), and succinic anhydride (2.2 g, 22.1 mmol) was added. The mixture was stirred overnight at room temperature, the reaction was concentrated and purified on a silica gel column, and dissociated with dichloromethane / methanol to give compound 53 (2.99 g, 84% yield). ESI-MS m / z: [M + H] + C 17H 29N 3O 8: Calculated 404.43; Found 404.11.

[0194] Example 48. Synthesis of bis((S)-4-ethyl-4-hydroxy-3,14-disideroxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-9-yl)4,4'-((((tert-butoxycarbonyl)azadiyl)bis(ethane-2,1-diyl))bis(azadiyl))bis(4-sideroxybutyrate) (54). Triethylamine (948 mg, 9.4 mmol) and HATU (1.79 g, 4.7 mmol) were added sequentially to a DMF (100 mL) solution of compound 53 (853 mg, 2.1 mmol) and compound 5 (1.71 g, 4.7 mmol). The resulting mixture was stirred overnight, then concentrated and purified by silica gel column chromatography (methanol / dichloromethane) to give compound 54 (2.84 g, 100% yield). ESI-MS m / z: [M + H] + C 57H 57N 7O 16: Calculated 1097.10; Found 1097.65.

[0195] Example 49. Synthesis of bis(((S)-4-ethyl-4-hydroxy-3,14-disideoxy-3,4,12,14-tetrahydro-1H-guanan-[3',4':6,7]indolo[1,2-b]quinoline-9-yl)4,4'-((azadiyl(ethane-2,1-diyl))bis(azadiyl))bis(4-sideoxybutyrate) (55). Compound 54 (2.84 g, 2.1 mmol) was dissolved in dichloromethane (40 mL), and trifluoroacetic acid (20 mL) was added. The reaction mixture was stirred at room temperature for 1 hour, and then concentrated to give compound 55 (3.3 g, 100% yield). ESI-MS m / z: [M + H] + C 52H 49N 7O 14: Calculated value 996.98; Found value 996.60.

[0196] Example 50. (S)-4-ethyl-4-hydroxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-9-yl 30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-37-(2-(4-( ...3',4':6,7]indolano[1,2-b]quinoline-9-yl 30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-37-(2-(4-((S)-4-ethyl-4-hydroxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[3',4':6,7]indolano[3',4':6,7]indolano[3',4':6,7]indolano[3',4':6,7]indolano[3',4':6,7]indolano[3',4':6,7]indolano[3',4 [',4':6,7]Indo[1,2-b]quinoline-9-yl)oxy)-4-sideoxybutyramine)ethyl)-27,31,36,41-tetra-sideoxy-2,5,8,11,14,17,20,23-octa-sideoxy-26,32,37,40-tetraazatetratetradecane-44-ester (56) Triethylamine (249 mg, 2.5 mmol) and HATU (234 mg, 0.60 mmol) were added sequentially to a DMF (20 mL) solution of compound 55 (614 mg, 0.60 mmol) and compound 53 (470 mg, 0.60 mmol). The resulting mixture was stirred for 40 min, then concentrated and purified by silica gel column chromatography (methanol / dichloromethane) to give compound 56 (46 mg, 5% yield). ESI-MS m / z: [M + H] + C 86H 105N 11O 28: Calculated 17410.81; Found 1742.01.

[0197] Example 51. Synthesis of (S)-4-ethyl-4-hydroxy-9-methoxy-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (57). 10-Hydroxycamptothecin (2.5 g, 6.86 mmol) was dissolved in DMF (150 mL), potassium carbonate (1.90 g, 13.72 mmol) and methyl iodide (1.17 g, 8.23 ​​mmol) were added, and the mixture was stirred overnight at room temperature. A mixture of petroleum ether (150 mL) and ethyl acetate (150 mL) was added to the reaction mixture and stirred. A yellow solid precipitated, which was collected by filtration and then dispersed in water (20 mL). 1 N hydrochloric acid was added dropwise until pH 7, and the mixture was filtered again to give compound 57 (1.0 g, 38% yield). ESI-MS m / z: [M + H] + C 21H 18N 2O 5 Calculated value 379.38; Found value 379.05.

[0198] Example 52. Synthesis of bis(((S)-4-ethyl-9-methoxy-3,14-di-sideoxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-4-yl)(((tert-butoxycarbonyl)azadiyl)bis(ethane-2,1-diyl))diaminocarbamate (59). Compound 57 (350 mg, 0.9 mmol), 4-dimethylaminopyridine (339 mg, 2.8 mmol), and triphosgene (93 mg, 0.34 mmol) were wet-milled under helium and mixed thoroughly. Then, anhydrous dichloromethane (8 mL) was added dropwise and the mixture was stirred for 10 minutes. Compound 52 (64 mg, 0.34 mmol) was dissolved in anhydrous dichloromethane (4 mL), and then triethylamine (93 mg, 0.9 mmol) was added. After stirring for 15 minutes, the solution was concentrated and purified by silica gel column chromatography (methanol / dichloromethane) to give compound 59 (200 mg, 22% yield). ESI-MS m / z: [M + H] +C 53H 53N 7O 14: Calculated value 1013.03; Found value 1013.26.

[0199] Example 53. Synthesis of bis(((S)-4-ethyl-9-methoxy-3,14-disideoxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-4-yl)(azabis(ethane-2,1-diyl))diaminocarbamate (60). Compound 59 (200 mg, 0.2 mmol) was dissolved in dichloromethane (10 mL) and reacted with trifluoroacetic acid (5 mL) for 4 hours. The reaction mixture was concentrated to give compound 60 (0.43 g, 100% yield). ESI-MS m / z: [M + H] + C 48H 45N 7O 12: Calculated value 912.91; Found value 912.62.

[0200] Example 54. Bis(((S)-4-ethyl-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-4-yl)((4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyryl)azadiyneyl)bis(ethane-2,1-diyl))diaminocarbamate (61) Triethylamine (112 μL, 0.81 mmol) and HATU (104 mg, 0.27 mmol) were added to a solution of compound 59 (249 mg, 0.27 mmol) and compound 7 (60 mg, 0.32 mmol) in dichloromethane (10 mL). The reaction was stirred for 40 min and then washed with water (20 mL). The organic phase was concentrated and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 61 (50 mg). ESI-MS m / z: [M + H] + C 56H 52N 8O 15 Calculated value 1078.06; Found value 1078.77.

[0201] Example 55. Synthesis of (S)-N,N'-(((((2S,20S)-11-(tert-butoxycarbonyl)-2,20-dimethyl-4,7,15,18-tetraoxy-3,8,11,14,19-pentaza-eicosico-1,21-diacetyl)bis(aza-diacetyl))bis(4,1-phenylene))bis(methylene))bis(1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-diacetyloxy-3,4,12,14-tetrahydro-1H-guanan-[3',4':6,7]indol[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaonium) (62). Compound 40 (96 mg, 0.132 mmol) and compound 53 (26 mg, 0.066 mmol) were dissolved in DMF (3 mL) and cooled to 0 °C. HATU (50 mg, 0.132 mmol) and N,NN-diisopropylethylamine (46 μL, 0.264 mmol) were then added, and the mixture was stirred at 0 °C for 30 min. The crude reaction mixture was purified directly by preparative HPLC (acetonitrile / water containing 0.1% formic acid) to give compound 62 (80 mg, 67% yield). ESI-MS m / z: [M] 2+C 91H 109F 2N 15O 18: Calculated 868.90; Found 868.90.

[0202] Example 56. Synthesis of (S)-N,N'-(((((2S,20S)-2,20-dimethyl-4,7,15,18-tetraoxy-3,8,11,14,19-pentazatetramono-1,21-diacetyl)bis(azadiyl)bis(4,1-phenylene))bis(methylene))bis(1-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanan[3',4';6,7]indolo[[1,2-b]quinolin-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaonium))(63). Compound 62 (80 mg, 0.043 mmol) was dissolved in a mixture of dichloromethane and trifluoroacetic acid (3 mL / 1 mL) and stirred at room temperature for 30 minutes. The reaction mixture was concentrated to give compound 63 (100% yield). ESI-MS m / z: [M] 2+C 86H 101F 2N 15O 16: Calculated 818.87; Found 818.87.

[0203] Example 57. (S)-N,N'-((((2S,20S)-11-((S)-30-(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-27,31-dioxy-2,5,8,11,14,17,20,23-octaoxa-26,32-diazahexadecane-36-acryl)-2,20-dimethyl-4,7 Synthesis of ,15,18-tetraoxy-3,8,11,14,19-pentaza-tetradecane-1,21-diacyl)bis(aza-diacyl))bis(4,1-phenylene))bis(methylene))bis(1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-diacyloxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinolin-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaonium)(64). At 0 °C, N,N-diisopropylethylamine (15 μL, 0.086 mmol) was added to a DMF (3 mL) solution of compound 63 (74 mg, 0.043 mmol) and compound 7 (39 mg, 0.0516 mmol), followed by heating to room temperature and stirring for 2 hours. After concentration, the residue was purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 64 (12 mg). ESI-MS m / z: [M+] 2+C 120H 157F 2N 19O 30: Calculated value 1191.06; Found value 1191.06.

[0204] Example 58. Synthesis of (S)-4-ethyl-8-fluoro-4,9-dihydroxy-11-methyl-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (103). 1-(2-amino-4-fluoro-5-hydroxyphenyl) ethyl ketone (0.41 g, 2.5 mmol) and compound 25 (0.62 g, 2.5 mmol) were dissolved in anhydrous toluene (40 mL), and p-toluenesulfonic acid (46 mg, 0.25 mmol) was added. The suspension was heated under reflux for 3 days and then cooled to room temperature. After removing the solvent, the residue was purified by column chromatography to give compound 103 (0.69 g, 73% yield) as a gray powder. ESI-MS m / z: [M + H] +C 21H 17FN 2O 5: Calculated value 397.11; Found value 397.16.

[0205] Example 59. Synthesis of (S)-9-(2-bromoethoxy)-4-ethyl-8-fluoro-4-hydroxy-11-methyl-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (104). A mixture of compound 103 (0.69 g, 1.74 mmol), anhydrous 1,2-dibromoethane (6.4 g, 34.8 mmol), and anhydrous potassium carbonate (1.2 g, 8.7 mmol) in anhydrous DMF (10 mL) was mechanically stirred for 16 hours at 80 °C. The reaction mixture was filtered through a diatomaceous earth mat, and the residue was thoroughly washed with DMF. The combined filtrate and washings were concentrated to dryness under vacuum to give a dark residue. The residue was purified by column chromatography (0-5% methanol / dichloromethane) to give compound 104 (0.74 g, 85%). ESI-MS m / z: [M + H] +C₂₃H₂₀BrFN₂O₅: Calculated 503.05; Found 503.05.

[0206] Example 60. Synthesis of (S)-9-(2-bromoethoxy)-4-ethyl-8-fluoro-4-hydroxy-11-methyl-10-nitro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (105). Compound 104 (0.74 g, 1.47 mmol) was slowly added to 1 mL of stirred concentrated sulfuric acid at 0 °C, and the resulting clear solution was cooled to -10 °C. A mixture of concentrated sulfuric acid (0.5 mL) pre-cooled to -10 °C and fuming nitric acid (0.5 mL) was added dropwise to the cooled reaction mixture at -10 °C. The reaction mixture was heated to 0 °C and stirred for 1 hour, then slowly poured onto ice chips. The yellow precipitate was filtered and washed with water, cold ethanol, and diethyl ether. The aqueous filtrate was filtered again through a diatomaceous earth mat, and the diatomaceous earth filter cake was extracted with 30% methanol / dichloromethane (50 mL). The organic solvent was concentrated to give another yellow solid. The combined yellow solids were slurried with ethanol to give compound 105 (0.74 g, 92%). ESI-MS m / z [M + H] +: C 23H 19BrFN 3O 7: Calculated value 548.04; Measured value 548.14.

[0207] Example 61. Synthesis of (S)-10-amino-9-(2-bromoethoxy)-4-ethyl-8-fluoro-4-hydroxy-11-methyl-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-3,14(4H,12H)-dione (106). Compound 105 (0.50 g, 0.91 mmol) was added in portions to a stirred concentrated hydrochloric acid solution (18 mL) at 0 °C. After 15 minutes, the resulting clear solution was cooled to -10 °C. Tin dioxide (0.86 g, 4.55 mmol) was then added in portions to the reaction mixture, and the mixture was warmed to room temperature. The mixture was stirred for 1.5 h and then slowly poured onto ice chips. The precipitate was filtered and washed with ethanol and ether, and the aqueous filtrate was extracted with 10% methanol / dichloromethane. The organic solution and the precipitate were combined and dissolved in 30% methanol / dichloromethane, then filtered through a short silicone mat and dissolved in 30% methanol / dichloromethane. The organic solvent was removed to give compound 106 (0.44 g, 94% yield), which could be used in the next step without further purification.

[0208] Example 62. Synthesis of (S)-9-ethyl-5-fluoro-9-hydroxy-16-methyl-2,3,12,15-tetrahydro-[1,4]oxazindo[3,2-f]guanano[3',4':6,7]indolo[1,2-b]quinoline-10,13(1H,9H)-dione (107). A solution of compound 106 (0.44 g, 0.85 mmol) in DMSO (4 mL) and sodium bicarbonate (0.10 g, 1.19 mmol) was stirred at 70 °C for 4 hours and diluted with hydrochloric acid (0.1 M, 8 mL) and water (40 mL). The precipitated solid was filtered, dissolved in a small amount of 10% methanol / dichloromethane, and purified by column chromatography using (1:20–1:6) methanol / dichloromethane as the solvent to give compound 107 (0.24 g, 66%). ESI-MS m / z: [M + H] + C 23H 20FN 3O 5: Calculated value 438.14; Found value 438.14.

[0209] Example 63. Synthesis of (S)-tertiary butyl (2-(9-ethyl-5-fluoro-9-hydroxy-16-methyl-10,13-di-side-oxy-2,3,9,10-tetrahydro-[1,4]oxazindo[3',4':6,7]indolo[1,2-b]quinoline-1(12H,13H,15H)-yl)ethyl)aminocarbamate (108). Sodium iodide (0.68 g, 4.56 mmol) and tributyl (2-chloroethyl) aminocarbamate (0.82 g, 4.56 mmol) were added to an anhydrous DMF (2 mL) stirred solution of compound 107 (0.20 g, 0.456 mmol). The mixture was heated at 120 °C for 18 hours. The reaction mixture was cooled to room temperature. It was concentrated to vacuum dryness and purified by column chromatography (0-5% methanol / dichloromethane) to give compound 108 (0.19 g, 75%). ESI-MS m / z: [M + H] +C 30H 33FN 4O 7: Calculated value 581.23; Found value 581.40.

[0210] Example 64. Synthesis of (S)-1-(2-aminoethyl)-9-ethyl-5-fluoro-9-hydroxy-16-methyl-2,3,12,15-tetrahydro-[1,4]oxazindo[3,2-f]guanano[3',4':6,7]indolo[1,2-b]quinoline-10,13(1H,9H)-dione (109). Add 2.5 mL of trifluoroacetic acid to a 5 mL solution of compound 108 (0.19 g, 0.327 mmol) in dichloromethane, and stir the reaction mixture at room temperature for 30 minutes. Concentrate the reaction mixture three times with dichloromethane to obtain compound 109, which can be used in the next step without further purification.

[0211] Example 65. Synthesis of compound 110. Compounds 109 and 7 (0.45 g, 0.49 mmol) obtained in the previous step were dissolved in DMF (5 mL), cooled to approximately 0 °C, and then N,N-diisopropylethylamine (172 μL, 0.98 mmol) was added. The reaction was heated to room temperature, stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 110 (359 mg, 60% yield). ESI-MS m / z: [M + H] +C 59H 81FN 8O 19: Calculated value 1224.56; Found value 1224.78.

[0212] Example 66. Synthesis of ditert-tert-butyl 4,4'-(((2R,3S)-2,3-bis(((benzyloxy)carbonyl)-amino)succinate)bis(azadiacryl))dibutyrate (172). To a DMA (70 mL) solution of compound 171 (4.25 g, 10.21 mmol), tributyl 4-aminobutyrate (3.25 g, 20.42 mmol), DMAP (4.47 g, 36.65 mmol), and EDC·hydrochloric acid (7.00 g, 36.65 mmol) were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column. The solution was dissociated with ethyl acetate / dichloromethane (1:10) to give compound 172 (6.50 g, 91% yield). ESI-MS m / z: [M + H] +C 36H 50N 4O 10 Calculated value 699.35; Found value 699.55.

[0213] Example 67. Synthesis of ditert-tert-butyl 4,4'-(((2R,3S)-2,3-diaminosuccinate)-bis(azadiacryl)) dibutyrate (173). A methanol (100 mL) solution of compound 172 (2.50 g, 3.58 mmol) was catalytically hydrogenated with 10% Pd / C (0.30 g, 50% wt) at room temperature for 18 hours. The Pd / C catalyst was then removed by diatomaceous earth filtration, and the filter cake was washed with methanol (~70 mL). The filtrate was concentrated to give compound 173, a yellow foam, which could be used in the next step without further purification (1.54 g, 100% yield). ESI-MS m / z: [M + H] + C 20H 38N 2O 6: Calculated 431.28; Found 431.50.

[0214] Example 68. Synthesis of ditert-tert-butyl 4,4'-((2R,3R)-2,3-bis(4-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)butyramine)succinyl)bis(azadiyl))dibutyrate (174). To a DMF (60 mL) solution of 4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyric acid (1.35 g, 7.39 mmol) and compound 173 (1.54 g, ~3.58 mmol), N,N-diisopropylethylamine (2.2 mL, 12.56 mmol) and HATU (4.77 g, 12.56 mmol) were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column, followed by dissociation with ethyl acetate / dichloromethane (1:10) to give the title compound (2.35 g, 86% yield). ESI-MS m / z [M + H] +: Calculated 761.36, Found 761.36.

[0215] Example 69.4 Synthesis of 4'-(((2R,3R)-2,3-bis(4-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)butyramine)succini)bis(azadiyl))dibutyric acid (175). Trifluoroacetic acid (10 mL) was added to a 20 mL mixture of compound 174 (2.30 g, 3.02 mmol) in dichloromethane. The mixture was stirred for 30 minutes, diluted with toluene (20 mL), and concentrated to give the title compound (1.69 g, 86% yield). ESI-MS m / z [M + H] +: C 28H 36N 6O 12 Calculated value 649.24, Found value 649.30.

[0216] Example 70. Synthesis of bis(2,5-dioxypyrrolidone-1-yl)4,4'-((2R,3R)-2,3-bis(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolidone)butyramine)succinyl)bis(azadiyl))dibutyrate (176). To a DMA (30 mL) solution of compound 175 (1.10 g, 1.69 mmol), N-hydroxysuccinimide (1-hydroxypyrrolidine-2,5-dione) (0.58 g, 5.08 mmol) and EDC·hydrochloric acid (1.25 g, 6.54 mmol) were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column. The solution was dissociated with ethyl acetate / dichloromethane (1:10) to give the title compound (1.30 g, 91% yield). ESI-MS m / z [M + H] +:C 36H 42N 8O 168 Calculated value 43.27, Found value 843.50.

[0217] Example 71. (S)-N,N'-((((2S,10S,11S,19S)-10,11-bis(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-2,19-dimethyl-4,9,12,17-tetraoxy-3,8,13,18-tetraazaeicosano-1,20-diacyl)bis(azadiyl))bis(4,1-phenylene))bis(methylene))bis(1-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,4,14-dioxy-3,4,12,14-tetrahydro-1H-guanan[3] Synthesis of [1,2-b]indolo[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaium (177). Compound 28 (94 mg, 0.12 mmol) and compound 176 (55 mg, 0.066 mmol) were dissolved in DMA (5 mL) and cooled to approximately 0 °C. Then, N,N-diisopropylethylamine (84 μL, 0.48 mmol) was added. The reaction was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 177 (23 mg, 19% yield). ESI-MS m / z: [M+] 2+C 106H 124F 2N 18O 22: Calculated value 1019.46; Found value 1019.50.

[0218] Example 72. Synthesis of 3-sideoxy-1-phenyl-2,7,10,13,16-penta-4-azanonadecan-19-acid (179). In a 500 mL flask, H₂N-PEG 4-CH₂CH₂CO₂H (3.0 g, 11.3 mmol, 1.0 eq) and potassium carbonate (4.7 g, 33.93 mmol, 3.0 eq) were dissolved in 50 mL of water and cooled on an ice-water bath. Benzyloxychloride (2.50 g, 14.7 mmol, 1.3 eq) was added dropwise to 50 mL of THF. The reaction mixture was heated to room temperature and stirred overnight. The pH of the reaction mixture was adjusted to 4–5 with 1N potassium sulfate, and the mixture was extracted with dichloromethane (200 mL × 1, 100 mL × 3), washed with water (500 mL) and brine (500 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was dissolved in a small amount of dichloromethane, then added to a silica gel column and dissolved with 2-4% methanol / dichloromethane. The fractions were combined and concentrated to give 3.8 g of a colorless oil (84% yield). ESI-MS m / z [M + H] +:C 19H 29NO 8 Calculated value 400.2, measured value 400.2.

[0219] Example 73.2 Synthesis of 5-dioxapyrrolidone-1-yl 3-epi-oxy-1-phenyl-2,7,10,13,16-pentaoxa-4-azanonadecan-19-ester. (180) To a solution of CbzHN-PEG 4-CH 2CH 2CO 2H (3.8 g, 9.5 mmol, 1.0 eq) in 50 mL of anhydrous dichloromethane, N-hydroxysuccinimide (1.3 g, 11.4 mmol, 1.2 eq) and EDC·hydrochloric acid (9.1 g, 47.5 mmol, 5.0 eq) were added. The reaction mixture was stirred overnight at room temperature, then washed with water (50 mL × 2) and brine (100 mL × 1), dried over anhydrous sodium sulfate, and concentrated. The crude product was used directly in the next step. ESI-MS m / z [M + H] +:C 23H 32N 2O 10 Calculated value 497.2, Measured value 497.2.

[0220] Example 74.3 Synthesis of 19-disideoxy-1-phenyl-2,7,10,13,16,23,26,29,32-nonoxa-4,20-diazapentadecane-35-acid (181). In a 300 mL flask, H₂N-PEG 4-CH₂CH₂CO₂H (2.6 g, 9.5 mmol, 1.0 eq) and potassium sulfate (3.9 g, 28.5 mmol, 3.0 eq) were dissolved in 40 mL of water. The mixture was cooled in an ice-water bath, and 40 mL of THF solution of the crude N-hydroxysuccinimide ester (3.8 g, 9.5 mmol) was added dropwise. The mixture was heated to room temperature and stirred overnight. The pH of the reaction mixture was adjusted to 4–5 with 1N potassium sulfate, and the mixture was extracted with dichloromethane (150 mL × 1, 100 mL × 2), washed with water (200 mL) and brine (200 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was dissolved in a small amount of dichloromethane and loaded onto a silicone column. It was then dissolved in 4-6% methanol / dichloromethane to obtain a colorless oil (4.91 g, 80% yield). ESI-MS m / z [M + H] +:C 30H 50N 2O 13 Calculated value 646.3, measured value 646.3.

[0221] Example 75. Synthesis of tributyl 3,19,35-trisyloxy-1-phenyl-2,7,10,13,16,23,26,29,32,39,42,45,48-tetraoxa-4,20,36-triazapentadecane-51-ester (182). H₂N-PEG 4-CH₂CH₂CO₂t Bu (0.48 g, 1.5 mmol, 1.0 eq) was dissolved in 3 mL of DMF and cooled on an ice / water bath. N,N-diisopropylethylamine (DIPEA) (0.78 g, 6.0 mmol, 4.0 eq) was added dropwise, followed by a DMF (7 mL) solution of compound 181 (0.97 g, 1.5 mmol, 1.0 eq) and HATU (1.72 g, 4.5 mmol, 3.0 eq). The reaction mixture was stirred on an ice bath for 2 hours, diluted with 100 mL of water, extracted with dichloromethane (100 mL × 3), washed with 1N potassium sulfate (200 mL), saturated sodium bicarbonate (200 mL), and brine (200 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was dissolved in a small amount of dichloromethane, loaded onto a silicone column, and dissociated with 0-5% methanol / dichloromethane. The fractions were combined and concentrated to give 1.22 g of a pale yellow oil (86% yield). ESI-MS m / z [M + H] +:C 45H 79N 3O 18 Calculated value 950.5, measured value 950.5.

[0222] Example 76. Synthesis of third butyl 1-amino-15,31-disideoxy-3,6,9,12,19,22,25,28,35,38,41,44-dodecoxa-16,32-diazatetrahexadecan-47-ester (183). A solution of compound 182 (1.22 g, 1.28 mmol) in dichloromethane (5 mL) was catalytically hydrogenated with Pd / C (5%, 500 mg) at room temperature (1 atm hydrogen) for 2 hours. The reactants were then filtered through diatomaceous earth, and the filter cake was washed with methanol. The filtrate and washings were combined and concentrated to give a pale yellow oil (1.04 g, 100% yield). ESI-MS m / z [M + H] +:C 37H 73N 3O 16 Calculated value 816.5, Found value 816.5.

[0223] Example 77. (50R,51R)-Ditert-butyl50,51-bis((benzyloxy)carbonyl)amino)-17,33,49,52,68,84-hexahedraloxy-4,7,10,13,20,23,26,29,36,39,42,45,56,59,62,65,72,75,78,81,88,91,94,97-tetracosoxa-16,32,48,53,69,85-hexaaza-1,100-diacetate (184) To a DMA (10 mL) solution of compound 171 (0.26 g, 0.64 mmol), a dichloromethane (5 mL) solution of compound 183 (1.04 g, 1.28 mmol) was added, followed by the addition of DMAP (0.23 g, 1.92 mmol) and EDC·hydrochloric acid (0.36 g, 1.92 mmol). The mixture was stirred overnight, concentrated, and purified on a silica gel column. The purified compound was dissociated with methanol / dichloromethane (1:10) to give compound 184 (0.81 g, 63% yield). ESI-MS m / z: [M+2H]2+C94H162N8O38 Calculated value 1006.55; Found value 1006.70.

[0224] Example 78. Synthesis of (50R,51R)-ditert-butyl-50,51-diamino-17,33,49,52,68,84-hexamoxy-4,7,10,13,20,23,26,29,36,39,42,45,56,59,62,65,72,75,78,81,88,91,94,97-tetracosoxa-16,32,48,53,69,85-hexaazaperoane-1,100-diacid ester (185). 10% Pd / C (100 mg, 5%) was added to a methanol (5 mL) solution of compound 184 (0.81 g, 0.40 mmol), and catalytic hydrogenation was carried out at room temperature for 18 hours. The Pd / C catalyst was then removed by diatomaceous earth filtration, and the filter cake was washed with methanol. The filtrate and washings were combined and concentrated to give compound 185 (0.70 g, 100% yield). ESI-MS m / z: [M+2H]2+C78H150N8O34: Calculated value 872.52; Found value 872.55.

[0225] Example 79. (50R,51R)-Ditert-tertiary butyl 50,51-bis(4-(2,5-disideloxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-17,33,49,52,68,84-hexahedraloxy-4,7,10,13,20,23,26,29,36,39,42,45,56,59,62,65,72,75,78,81,91,94,97-tetracosoxa-16,32,48,53,69,85-hexaazaperoane-1,100-diacid ester (18 6) Synthesis of (50S,51S)-50,51-bis(4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-17,33,49,52,68,84-hexahedraloxy-4,7,10,13,20,23,26,29,36,39,42,45,56,59,62,65,72,75,78,81,88,91,94,97-tetracosoxa-16,32,48,53,69,85-hexaazaperoane-1,100-diacid (187). To a DMF (5 mL) solution of 4-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)butyric acid (0.17 g, 1.00 mmol) and compound 185 (0.70 g, 0.40 mmol), N,N-diisopropylethylamine (0.88 mL, 5 mmol) and HATU (1.90 g, 12.56 mmol) were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column. The solution was dissociated with 1–10% methanol / dichloromethane to give compound 186 (0.548 g, 66% yield) as an oil. ESI-MS m / z [M+2H]²⁺: C 94H 166N 10O 40 Calculated value 2075.1264, Found value 2075.1350. Compound 186 (0.54 g, 0.26 mmol) was dissolved in dichloromethane (5 mL) and reacted with trifluoroacetic acid (2.5 mL). The mixture was stirred at room temperature, diluted with toluene (20 mL) for 30 minutes, and concentrated to give the title compound 187 (0.488, 96% yield), which could be used in the next step without further purification. ESI-MS m / z [M+H]+: C 86H 149N 10O 40 Calculated value 1961.9933, Found value 1961.9987.

[0226] Example 80. (S)-N,N'-((((2S,53S,54S,105S)-53,54-bis(4-(2,5-disideloxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-2,105-dimethyl-4,20,36,52,55,71,87,103-octasideloxy-7,10,13,16,23,26,29,32,39,42,45,48,59,62,65,6 8,75,78,81,84,91,94,97,100-tetracosoxa-3,19,35,51,56,72,88,104-octaazahexadecane-1,106-diacyl)bis(azadiacyl))bis(4,1-phenylene))bis(methylene))bis(1-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-disideloxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaonium)(188) Compound 28 (47 mg, 0.060 mmol) and compound 187 (59 mg, 0.030 mmol) were dissolved in DMA (5 mL), cooled to approximately 0 °C, and then dissolved in N,N-diisopropylethylamine (21 μL, 0.12 mmol). The reaction was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 188 (36 mg, 36% yield). ESI-MS m / z: [M+] 2+C 164H 238F 2N 22O 50: Calculated value 1675.8279; Found value 1675.8392.

[0227] Example 81. 2,5,8,11,14,17,20,23,26-Nonoxaoctacosane-28-tertiary butyl ester (191) Sodium hydride (60%, 24 g, 600 mmol) was added to a solution of octaethylene glycol monomethyl ether (115 g, 300 mmol) in THF (3.0 L). The mixture was stirred at room temperature for 1 hour. Then, tributyl 2-bromoacetate (146 g, 750 mmol) was added to the mixture and stirred at room temperature for 1 hour. The mixture was then diluted with dichloromethane (4 L) and poured into ice water (2 kg). The organic phase was separated, and the aqueous phase was extracted with dichloromethane (1 L). The combined organic phases were washed with water and dried over anhydrous sodium sulfate. Purification by column chromatography (20% ethyl acetate / petroleum ether, then purified dichloromethane to 5% methanol / dichloromethane) yielded the title compound (108 g, 72% yield) as a yellow oil.

[0228] Example 82.2,5,8,11,14,17,20,23,26-Nineoxaoctacosane-28-acid (192) Synthesis. 2,5,8,11,14,17,20,23,26-nonoxaoctacosane-28-tert-butyl ester (210 g, 422 mmol) was dissolved in dichloromethane (400 mL) and anhydrous formic acid (1 L). The resulting solution was stirred overnight at room temperature. All volatiles were removed under vacuum to give the title compound (200 g, >100% yield) as a yellow oil.

[0229] Example 83.2,5,8,11,14,17,20,23,26-Nineoxaoctadecane-28-acetyl chloride (193) synthesis. To a solution of 2,5,8,11,14,17,20,23,26-nonoxaoctacosan-28-acid (198 g, 422 mmol) in dichloromethane (2.6 L), oxalic acid (275 mL) and DMF (0.5 mL) were added at room temperature. The resulting solution was stirred continuously at room temperature for 3 hours. All volatiles were removed under vacuum to give the title compound (210 g, >100% yield) as a yellow oil.

[0230] Example 84. Synthesis of (S)-34-((benzyloxy)carbonyl)amino)-28-sideoxy-2,5,8,11,14,17,20,23,26-nonoxa-29-azapentadecane-35-acid (194). ZL-Lys-OH (236 g, 844 mmol), sodium carbonate (89.5 g, 844 mmol), and sodium hydroxide (33.8 g, 844 mmol) were dissolved in water (1.6 L). The mixture was cooled to 0 °C using an ice-salt bath, and a THF solution of 2,5,8,11,14,17,20,23,26-nonoxaoctacosane-28-acetylchloro(210 g, 422 mmol) in 160 mL was added. The resulting mixture was stirred continuously at room temperature for 1 hour, and then diluted with ethyl acetate (1 L). The aqueous layer was separated, and concentrated hydrochloric acid was added under ice cooling until pH 3 was reached. The mixture was extracted with dichloromethane, the organic layer was washed with brine, dried over sodium sulfate, and concentrated to give the title compound (290 g, 97% yield) as a yellow oil.

[0231] Example 85. Synthesis of (S)-pentafluorophenyl 34-((((benzyloxy)carbonyl)amino)-28-sideoxy-2,5,8,11,14,17,20,23,26-nonasideoxy-29-azapentadecane-35-ester (195). To a 2 L solution of compound 194 (183 g, 260 mmol) in dichloromethane, pentafluorophenol (95.4 g, 520 mmol) and DIC (131 g, 1.04 mol) were added. The reaction mixture was stirred at room temperature for 1 hour, and then concentrated to give the crude product (430 g). Example 86. Synthesis of (S)-tert-butyl 34-((benzyloxy)carbonyl)amino)-28,35-disideoxy-2,5,8,11,14,17,20,23,26-nonoxa-29,36-diazatezoracan-40-ester (196). At 0 °C, N,N-diisopropylethylamine (134 g, 1.04 mol) was added to a DMF (1.5 L) solution of 62.0 g (390 mmol) of tributyl 4-aminobutyrate. Then, compound 195 (430 g, crude product) was added at 10–20 °C, and the resulting mixture was stirred at room temperature for 1 hour. The DMF was removed under vacuum, and the residue was diluted with dichloromethane and washed with water. The aqueous phase was back-extracted with dichloromethane. The combined organic phases were washed with 0.2 N hydrochloric acid and brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The title compound (180 g, 82% yield) was purified by column chromatography (25% ethyl acetate / PE to pure ethyl acetate, then 0–5% methanol / dichloromethane) as a yellow oil.

[0232] Example 87. Synthesis of (S)-tert-butyl34-amino-28,35-disideoxy-2,5,8,11,14,17,20,23,26-nonasideoxy-29,36-diazatracetradecane-40-ester (197). A methanol (500 mL) solution of compound 196 (78.0 g, 92.3 mmol, 1.0 eq) and Pd / C (13 g, 10% Pd / C, 50% wet) were catalytically hydrogenated overnight at room temperature (1 atm hydrogen). The mixture was then filtered and concentrated. The residue was purified by column chromatography (0 to 20% methanol / dichloromethane) to give the title compound (70.2 g, 92% yield) as a pale yellow oil.

[0233] Example 88. Synthesis of (7S,10R,11R,14S)-ditert-butyl10,11-bis((((benzyloxy)carbonyl)amino)-6,9,12,15-tetraoxy-7,14-bis(28-sideoxy-2,5,8,11,14,17,20,23,26-nonasideoxy-29-azatritriatrisane-33-yl)-5,8,13,16-tetraazaeicosane-1,20-diacid ester (198). [] [] To a DMA (10 mL) solution of compound 171 (0.85 g, 2.00 mmol), a dichloromethane (10 mL) solution of compound 197 (3.20 g, 4.50 mmol), a DMAP (1.50 g, 12 mmol) solution, and an EDC·hydrochloric acid (2.3 g, 12 mmol) solution were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column. The purified compound was dissociated with ethyl acetate / dichloromethane (1:10) to give compound 198 (3.33 g, 88% yield). ESI-MS m / z: [M+2H]2+C86H146N8O32 Calculated value 902.50; Found value 902.55.

[0234] Example 89. Synthesis of (7S,10R,11R,14S)-ditert-butyl10,11-bis(4-(2,5-disideoxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-6,9,12,15-tetrasideoxy-7,14-bis(28-sideoxy-2,5,8,11,14,17,20,23,26-nonoxa-29-azatritriatriane-33-yl)-5,8,13,16-tetraazaeicosane-1,20-diester (199). [] A mixture of compound 198 (3.33 g, 1.76 mmol), Pd / C (5 wt%, 0.10 g), and dichloromethane (50 mL) was catalytically hydrogenated overnight (1 atm hydrogen), and then filtered on diatomaceous earth. The filtrate was concentrated and then dissolved in DMF (10 mL), and EDC (1.00 g, 5.28 mmol) and compound 4 (1.84 g, 5.28 mmol) were added. The mixture was stirred at room temperature for 16 hours, concentrated, and purified by silica gel column chromatography (1:4 methanol / dichloromethane) to give an oil (2.56 g, 78% yield). ESI-MS m / z: [M+2H]2+C86H148N10O34 Calculated value 933.51; Found value 933.55.

[0235] Example 90. (S)-N,N'-(((((2S,10S,13R,14R,17S,25S)-13,14-bis(4-(2,5-disideloxy-2,5-dihydro-1H-pyrrolo-1-yl)butyramine)-2,25-dimethyl-4,9,12,15,18,23-hexamyloxy-10,17-bis(28-sideloxy- Synthesis of 2,5,8,11,14,17,20,23,26-nonazooxy-29-azatritane-33-yl)-3,8,11,16,19,24-hexaazahexacosane-1,26-bis(azadiyl))bis(1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indol[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-amino)carbamate (200). A mixture of compound 199 (1.00 g, 0.536 mmol) in dichloromethane (5 mL) and formic acid (5 mL) was stirred at room temperature for 24 hours, then concentrated. The residue was dissolved in DMA (5 mL), and compound 28 (0.64 g, 0.89 mmol), triethylamine (0.15 mL, 1.07 mmol), and HATU (0.41 g, 1.07 mmol) were added, followed by stirring at room temperature for 16 hours. After solvent removal under high vacuum, the residue was purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 200 (1.06 g, 63% yield). ESI-MS m / z: M 2+C 156H 220F 2N 22O 44 Calculated value 1571.78; Found value 1571.78.

[0236] Example 91. Synthesis of methyl 4-(bis(2-hydroxyethyl)amino)-4-sideoxybutyrate (202). [] Dimethyl succinate (20.0 g, 136.9 mmol) and dihydroxyethylamine (7.20 g, 68.7 mmol) were heated in a mixture of anhydrous toluene (500 mL) and pyridine (50 mL) at 150 °C for 28 h. The mixture was concentrated and purified on a silica column, dissociated with 5–25% ethyl acetate / dichloromethane, to give the title compound (12.5 g, 83% yield). ESI-MS m / z 242.42 ([M + Na] +).

[0237] Example 92.4 Synthesis of methyl bis(2-(((methanesulfonyl)oxy)ethyl)amino)-4-sideoxybutyrate (203). [] Methyl 4-(bis(2-hydroxyethyl)amino)-4-sideoxybutyrate (12.0 g, 49.56 mmol) was added to a solution of anhydrous pyridine (350 mL) with methanesulfonyl chloride (20.0 g, 175.4 mmol). After stirring overnight, the mixture was concentrated, diluted with ethyl acetate (350 mL), washed with cold 1 M sodium dihydrogen phosphate (2 × 300 mL), dried over sodium sulfate, filtered, and concentrated to give the crude product (18.8 g, >100% yield). The crude product was ready for the next step without further purification. ESI-MS m / z 376.06 ([M + H] +).

[0238] Example 93.3 Synthesis of 6-endo-oxy-Δ-tetrahydrophthalimide (204). [] Furan (10.0 mL, 137.4 mmol) was added to a 200 mL solution of maleimide (10.0 g, 103.0 mmol) in toluene. The mixture was heated in a 1 L reaction vessel at 100 °C for 8 hours, cooled to room temperature, washed with methanol, concentrated, and crystallized in ethyl acetate / n-hexane to give 16.7 g (99%) of the title compound. ¹H NMR (CDCl₃): 11.12 (s, ¹H), 6.68–6.64 (m, 2H), 5.18–5.13 (m, 2H), 2.97–2.92 (m, 2H); ESI-MS m / z 188.04 ([M + Na]⁺).

[0239] Example 94.4 Synthesis of methyl 2-((2-((3aR,4R,7S,7aS)-1,3-disideloxy-3a,4,7,7a-tetrahydro-1H-4,7-epoxyisoindole-2(3H)-yl)ethyl)(2-(((4R,7S,7aS)-1,3-disideloxy-3a,4,7,7a-tetrahydro-1H-4,7-epoxyisoindole-2(3H)-yl)ethyl)amino)-4-sideloxybutyrate (205). To a DMA (350 mL) solution of methyl 4-(bis(2-((((methanesulfonyl)oxy)ethyl)amino)-4-sideoxybutyrate (203, freshly prepared, 90% purity, 8.5 g, 20 mmol), 3,6-endosideoxy-Δ-tetrahydrophthalimide (204, 10.2 g, 61.8 mmol), sodium carbonate (8.0 g, 75.5 mmol), and sodium iodide (0.3 g, 2.0 mmol) were added. The mixture was stirred overnight at room temperature, concentrated, diluted with ethyl acetate (350 mL), and then diluted with saturated sodium bicarbonate solution (300 mL), saline solution (300 mL), and 1 M potassium dihydrogen phosphate solution (300 mL). The organic layer was washed with sodium sulfate, filtered, concentrated, loaded onto a silica column, and dissolved in 10-30% ethyl acetate / n-hexane to give the title compound (7.9 g, 77% yield). ESI-MS m / z 536.4 ([M+Na]+).

[0240] Example 95.4 Synthesis of bis(2-(2-,2,5-disideroxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-sideroxybutyric acid. (206) [] A solution of compound 205 (3.0 g, 5.8 mmol) and trimethyltinanol (4.8 g, 26.4 mmol) in 1,2-dichloroethane (150 mL) was refluxed at 80 °C for 8 hours, then cooled to room temperature. The residue was passed through a short silica column and dissociated with dichloromethane / methanol to remove excess trimethyltin hydroxide. The fractions were then combined, concentrated, and diluted with DMA and toluene. The mixture was heated to 120 °C and stirred overnight. The reaction mixture was loaded onto a silica column and dissociated with 5–10% methanol / dichloromethane to give the title compound (1.62 g, 76% yield). ESI-MS m / z 386.2 ([M+Na]+).

[0241] Example 96.2 Synthesis of 5-dioxapyrrolidine-1-yl 4-(bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-sideoxybutyrate (207). To a 10 mL DMA solution of compound 206 (1.62 g, 4.46 mmol, 1.0 eq), N-hydroxysuccinimide (0.61 g, 5.35 mmol, 1.2 eq) and EDC·hydrochloric acid (1.71 g, 8.92 mmol, 2.0 eq) were added. The reaction mixture was stirred overnight at room temperature, then washed with water (50 mL × 2) and brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to give an oil (2.00 g). The crude product was used directly in the next step. ESI-MS m / z [M + H] +:C 20H 20N 4O 9 Calculated value 461.12, Found value 461.24.

[0242] Example 97. Synthesis of N-(4-((S)-2-(4-(bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-dioxybutyramine)propamidamine)benzyl)-1-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaium (208). At 0 °C, the crude product from the previous step (0.20 g) was dissolved in DMA (5 mL), compound 28 (0.71 g, 1.00 mmol) and N,N-diisopropylethylamine (0.20 mL, 1.20 mmol) were added. The reaction mixture was heated to room temperature, stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 208 (0.85 g, 80% yield). ESI-MS m / z: M +C 55H 61FN 9O 12: Calculated value 1058.44; Found value 1058.60.

[0243] Example 98. Synthesis of (S)-tertiary butyl 34-(4-(bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-dioxybutyramine)-28,35-dioxy-2,5,8,11,14,17,20,23,26-nonoxa-29,36-diazatoracolide-40-ester (210). To a DMA (20 mL) solution of compound 197 (2.98 g, 4.20 mmol) and compound 206 (1.39 g, 3.82 mmol), EDC·hydrochloric acid (0.80 g, 4.20 mmol) was added. The reaction mixture was stirred overnight at room temperature, then poured into water (50 mL) and extracted with ethyl acetate (3 × 40 mL). The combined organic phases were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (10–50% ethyl acetate / petroleum ether) to give a colorless oil (3.23 g, 80% yield). ESI-MS m / z 1057.85 ([M + H] +).

[0244] Example 99. Synthesis of (S)-34-(4-(bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-dioxybutyramine)-28,35-dioxy-2,5,8,11,14,17,20,23,26-nonoxa-29,36-diazatoracoacetane-40-acid (211). A solution of compound 210 (3.20 g, 3.03 mmol) in formic acid (10 mL) and dichloromethane (5 mL) was stirred overnight at room temperature. The solution was then concentrated and concentrated three times with toluene to give a colorless oil (3.00 g, crude product), which was ready for use without further purification. ESI-MS m / z 1001.50 ([M + H] +).

[0245] Example 100. Synthesis of (S)-2,5-dioxyrrolidine-1-yl 34-(4-(bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-dioxybutyramine)-28,35-dioxy-2,5,8,11,14,17,20,23,26-nonoxa-29,36-diazattracosan-40-ester (212). To a DMA (15.0 mL) solution of compound 211 (3.00 g, crude product, 3.03 mmol), N-hydroxysuccinimide (0.38 g, 3.33 mmol) and EDC·hydrochloric acid (0.87 g, 4.55 mmol) were added. The mixture was stirred continuously at room temperature for 2 hours, then diluted with water (50 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic phases were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (10–50% ethyl acetate / petroleum ether) to give a colorless oil (2.90 g, 90% yield). ESI-MS m / z 1098.50 ([M + H] +).

[0246] Example 101. N-(4-((34S,42S)-34-(4-(bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)-4-dioxybutyramine)-42-methyl-28,35,40-trioxy-2,5,8,11,14,17,20,23,26-nonoxa-29,36,41-triazatetrazotrialkylamino)benzyl)-1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-pirano[3] [',4':6,7]Indo[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaium(213) Compound 212 (0.10 g, 0.091 mmol) was dissolved in DMA (5 mL) at 0 °C, and compound 28 (56.8 mg, 0.08 mmol) and N,N-diisopropylethylamine (0.020 mL, 0.12 mmol) were added. The reaction mixture was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 213 (84 mg, 62% yield). ESI-MS m / z: M +C 84H 116FN 12O 24: Calculated value 1695.82; Found value 1695.82.

[0247] Example 102.2 Synthesis of tert-butyl ester (216) of (2-(1,3-dioxaindoline-2-yl)acetyl)hydrazinocarboxylic acid. [] [] At 0 °C, triethylamine (13.5 mL, 97.4 mmol) and compound 215 (10.8 g, 48.7 mmol) were added sequentially to a solution of Boc-hydrazine (7.08 g, 53.5 mmol) in dichloromethane (200 mL). The mixture was stirred at room temperature for 30 minutes, poured into ice water (100 mL), and extracted with dichloromethane (3 × 100 mL). The combined organic phases were washed with water (100 mL) and brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid (15.5 g, 100% yield). ESI-MS m / z 320.12 ([M + H] +).

[0248] Example 103.2 Synthesis of (1,3-dioxoisoindol-2-yl)acetylhydrazine. (217) [] [] Compound 216 (15.5 g, 48.7 mmol) was dissolved in 1,4-dioxane (150 mL) and reacted with 25% hydrochloric acid (50 mL) at room temperature for 1 hour. The reaction mixture was concentrated and then co-concentrated with toluene to give a white solid (10.6 g, 100% yield). ESI-MS m / z 220.06 ([M + H] +).

[0249] Example 104.2 Synthesis of (1,3-dioxaindololin-2-yl)-N'-(2-(1,3-dioxaindololin-2-yl)acetyl)acetylhydrazine (218). [] [] At 0 °C, triethylamine (13.5 mL, 97.4 mmol) and compound 215 (10.8 g, 48.7 mmol) were added to a THF (200 mL) solution of compound 217 (10.6 g, 48.7 mmol). The reaction was heated to room temperature and stirred overnight. The precipitate was collected by filtration and then suspended in water (100 mL) and stirred for 20 minutes. The mixture was filtered again to give compound 218 (15.7 g, 80% yield) as a white solid. ESI-MS m / z 407.09 ([M + H] +).

[0250] Example 105. Synthesis of ditert-tert-butyl 2,2'-(1,2-bis(2-(1,3-dioxaindololin-2-yl)acetyl)hydrazine-1,2-diyl)diacetate (219). [] [] At 0 °C, sodium hydride (0.5 g, 12.3 mmol) was added in portions to a DMF (40 mL) solution of compound 218 (2.0 g, 4.92 mmol). The mixture was heated to room temperature and stirred for 3 hours. Tributyl bromoacetate (2.0 g, 10.3 mmol) was added, and the reaction was stirred overnight. The mixture was then poured into ice water (100 mL) and extracted with dichloromethane (3 × 50 mL). The combined organic phases were washed with water (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel chromatography to give a white solid (1.5 g, 50% yield). ESI-MS m / z 635.23 ([M + H] +).

[0251] Example 106. Synthesis of di-tert-butyl 2,2'-(1,2-bis(2-aminoacetyl)hydrazine-1,2-diyl)diacetate (220). [] [] The mixture of compound 219 (1.5 g, 2.36 mmol) and hydrazine (442 mg, 7.08 mmol) in ethanol (30 mL) was refluxed for 1 hour, then cooled to room temperature and filtered. The filtrate was concentrated and added to ethyl acetate (20 mL), and filtered again. The filtrate was concentrated to give a white solid 220 (750 mg, 85% yield). ESI-MS m / z 375.22 ([M + H]+).

[0252] Example 107. Synthesis of ditert-tert-butyl 2,2'-(1,2-bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)acetyl)hydrazine-1,2-diyl)diacetate (221). [] [] At 0 °C, N-methoxycarbonylmaleimide (622 mg, 4 mmol) was added to a mixture of 20 mL THF and 30 mL saturated sodium bicarbonate aqueous solution of compound 220 (750 mg, 2 mmol). The reaction mixture was stirred at 0 °C for 1 hour. The white solid was collected by filtration to give compound 221 (854 mg, 80% yield). ESI-MS m / z 535.20 ([M + H] +).

[0253] Example 108.2 Synthesis of 2'-(1,2-bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)acetyl)hydrazine-1,2-diyl)diacetic acid (222). [] [] Compound 221 (854 mg, 1.6 mmol) was dissolved in dioxane (3 mL) and reacted with 25% hydrochloric acid (3 mL) at room temperature for 2 hours. The reaction mixture was then concentrated to give compound 222 (675 mg, 100% yield). ESI-MS m / z 423.07 ([M + H] +).

[0254] Example 109. Synthesis of ditert-tert-butyl 4,4'-((2,2'-(1,2-bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)acetyl)hydrazine-1,2-diyl)bis(acetyl))bis(azadiyl))dibutyrate (223). [] [] At 0 °C, 4-aminobutyric acid tributyl ester (158 mg, 0.99 mmol) and EDC hydrochloric acid (189.7 mg, 0.99 mmol) were added to a DMF (5 mL) solution of compound 222 (200 mg, 0.47 mmol). The reaction mixture was heated to room temperature and stirred overnight, then poured into ice water and extracted with dichloromethane (3 × 10 mL). The combined organic phases were washed with 0.2 N hydrochloric acid (5 mL), water (5 mL), and brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid (330 mg, 100% yield).

[0255] Example 110. Synthesis of bis(2,5-dioxopyrrolidone-1-yl)4,4'-((2,2'-(1,2-bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)acetyl)hydrazine-1,2-bis(acetyl))bis(azadiacetyl))dibutyrate (225). [] [] Compound 223 (330 mg, 0.47 mmol) was dissolved in dioxane (3 mL) and reacted with 25% hydrochloric acid (3 mL) at room temperature for 2 hours. The reaction mixture was concentrated and redissolved in DMF (5 mL), cooled to 0 °C, and N-hydroxysuccinimide (113 mg, 0.98 mmol) and EDC·hydrochloric acid (189 mg, 0.98 mmol) were added sequentially. The reaction mixture was heated to room temperature and stirred overnight, poured into ice water, and extracted with dichloromethane (3 × 20 mL). The combined organic phases were washed with water (5 mL) and brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid 225 (369 mg, 100% yield). ESI-MS m / z 787.21 ([M + H] +).

[0256] Example 111. (S)-N,N'-((((((2S,21S)-11,12-bis(2-(2,5-dioxy-2,5-dihydro-1H-pyrrolo-1-yl)acetyl)-2,21-dimethyl-4,9,14,19-tetraoxy-3,8,11,12,15,20-hexaazadocosane-1,22-diacetyl)bis(azadiacetyl))bis(4,1-phenylene))bis(methylene))bis(1-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-pirano[3 Synthesis of [',4':6,7]indo[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-amino)226. Compound 225 (31.5 mg, 0.04 mmol) was dissolved in DMA (5 mL) at 0 °C, and compound 28 (56.8 mg, 0.08 mmol) and N,N-diisopropylethylamine (0.020 mL, 0.12 mmol) were added. The reaction mixture was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 226 (57 mg, 72% yield). ESI-MS m / z: [M+] 2+C 102H 116F 2N 18O 2: Calculated value 991.42; Found value 991.86.

[0257] Example 112. Synthesis of third butyl 3-((2-aminoethyl)amino)propionate (228). A solution of tributyl acrylate (12.81 g, 0.10 mmol) and ethane-1,2-diamine (24.3 g, 0.40 mol) in THF (150 mL) was stirred for 24 hours at 45 °C. The mixture was concentrated and purified on an alumina column, followed by dissociation with triethylamine / methanol / dichloromethane (5:15:80) to give the title compound (17.50 g, 92% yield). ESI-MS m / z 189.20 ([M+H]+).

[0258] Example 113. Synthesis of (2-aminoethyl)amino)propionate (229). A solution of 1,4-dioxane (50 mL) of tert-butyl 3-(((2-aminoethyl)amino)propionate (17.00 g, 90.33 mmol) was reacted with concentrated hydrochloric acid (15 mL). The mixture was stirred at room temperature for 30 min, concentrated, and diluted with purified water (150 mL) and ethyl acetate / petroleum ether (40 mL, 1:5). The mixture was separated, and the organic layer was extracted with water (2 × 10 mL). The aqueous layer was concentrated and dried under vacuum to give the title compound (18.70 g, 100% yield, LC-MS purity 96%). ESI-MS m / z 133.20 ([M + H]+).

[0259] Example 114.3 Synthesis of ((2-(2,5-dioxy-2,5-dihydro-1H-pyrrole-1-yl)ethyl)amino)propionic acid (230). Maleic anhydride (8.85 g, 90.33 mmol) was added to a THF (150 mL) solution of 3-((2-aminoethyl)amino)propionic acid (18.70 g, 90.33 mmol) at 0 °C. The mixture was stirred at 0–4 °C for 4 hours and concentrated to give a quantitative yield of (Z)-4-((2-(((2-carboxyethyl)amino)ethyl)amino)-4-sideoxybut-2-enoic acid, confirmed by LC-MS. Toluene (150 mL) and DMA (50 mL) were added at 90 °C, and the mixture was refluxed using a Dean-Stark separatory. After collecting 30 mL of solvent, hexamethyl silane (9.0 mL, 43.15 mmol) and zinc chloride (16 mL, 1.0 M ether solution) were added. The mixture was heated to 115–125 °C, and toluene was collected using a Dean-Stark separatory. The reaction mixture was refluxed at 120 °C for 6 hours. During this time, 2 × 40 mL of anhydrous toluene was added to maintain the mixture volume at approximately 50 mL. The mixture was then cooled, and 1 mL of a 1:10 solution of concentrated hydrochloric acid / methanol was added. The mixture was concentrated and purified on a silica gel column using water / The compound was dissolved in methanol (1:15) and dried under vacuum to give 14.75 g of the title compound (77.0% yield). ESI-MS m / z 213.10 ([M+H]+).

[0260] Example 115. Synthesis of 2,5,8,11,14,17,20,23-octa-sideoxytetradecane-25-yl 4-methylbenzenesulfonate (231). TsCl (30.2 g, 0.159 mol) was added to a solution of 2,5,8,11,14,17,20,23-octa-semi-oxytetradecano-25-ol (50.0 g, 0.130 mol) in dichloromethane (200 mL) and pyridine (100 mL). The mixture was stirred overnight, concentrated, and purified on a silica gel column. The solution was dissociated with acetone / dichloromethane (1:1 to 4:1), concentrated, and dried under vacuum to give 57.34 g (82.0% yield) of the title compound. ESI-MS m / z 539.40 ([M+H]+).

[0261] Example 116. Synthesis of S-2,5,8,11,14,17,20,23-octa-sideoxytetradecane-25-ethylethylsulfate (232). To a mixed solution (50 mL) of 2,5,8,11,14,17,20,23-octa-semi-oxytetradecano-25-yl-4-methylbenzenesulfonate (57.30 g, 0.106 mol) in THF (300 mL) and N,N-diisopropylethylamine, HSAc (10.0 g, 0.131 mol) was added. The mixture was stirred overnight, concentrated, and purified on a silica gel column. The solution was dissociated with ethyl acetate / dichloromethane (1:2 to 4:1), concentrated, and dried under vacuum to give 40.51 g (86% yield) of the title compound. ESI-MS m / z 443.35 ([M+H]+).

[0262] Example 117. Synthesis of 2,5,8,11,14,17,20,23-octa-sideoxytetradecane-25-sulfonic acid (233). S-2,5,8,11,14,17,20,23-octa-sideoxytetradecane-25-ethyl ethimide (40.40 g, 0.091 mol) was added to a mixture of acetic acid (200 mL) and 30% hydrogen peroxide (100 mL) and stirred overnight. The mixture was concentrated, diluted with pure water (200 mL) and toluene (150 mL), separated, and the organic layer was extracted with water (2 × 25 mL). The aqueous solutions were combined, evaporated, and dried under vacuum to give 40.50 g of the title compound (99% yield, LC-MS purity 95%). ESI-MS m / z 449.30 ([M + H]+).

[0263] Example 118. Synthesis of 3,3-N,N-(2”-maleiminoethyl)(2’,5’,8’,11’,14’,17’,20’,23’,26’-nonoxaoctacosane-28’-thionine)aminopropionic acid (234). To a mixed solution of 2,5,8,11,14,17,20,23-octa-sideoxy-pecosane-25-sulfonic acid (20.0 g, 44.62 mmol) in THF (100 mL) and dichloromethane (100 mL), oxalic acid (25.21 g, 200.19 mmol) and DMF (0.015 mL) were added sequentially. The mixture was stirred at room temperature for 2 hours, concentrated, concentrated with dichloromethane / toluene (1:1, 2 × 50 mL), and then dissolved in THF (50 mL). To a THF (100 mL) solution of 3-((2-(2,5-di-sideoxy-2,5-dihydro-1H-pyrrolo-1-yl)ethyl)amino)propionic acid compound (7.50 g, 35.36 mmol), the above-mentioned sulfonic acid was added. The mixture was stirred overnight, concentrated, and purified on a silica column. It was then dissociated with methanol / dichloromethane (1:6 to 1:5), concentrated, and dried under vacuum to give 14.76 g (65% yield) of the title compound. ESI-MS m / z 643.35 ([M+H]+).

[0264] Example 119. Synthesis of NN-succinimide 3,3-N,N-(2''-maleimide ethyl)(2',5',8',11',14',17',20',23',26'-nonoxaoctacosane-28'-thionine) aminopropionate (235). A mixture of 3,3-N,N-(2”-maleiminoethyl)(2’,5’,8’,11’,14’,17’,20’,23’,26’-nonoxaoctacosane-28’-thionine)aminopropionic acid (234) (7.50 g, 11.67 mmol), N-hydroxysuccinimide (1.50 g, 13.04 mmol), EDC hydrochloric acid (10.10 g, 52.60 mmol), and THF (100 mL) was stirred overnight, concentrated under vacuum, purified on a silica gel column, dissociated with ethyl acetate / dichloromethane (1:4 to 2:1), concentrated, and dried under vacuum to give 6.30 g (73% yield) of the title compound. ESI-MS m / z 740.40 ([M+H]+).

[0265] Example 120. Synthesis of compound 236. [] At 0 °C, N,NN-diisopropylethylamine (3 mL) was added to a DMF (15 mL) solution of H-Gly-Gly-Gly-OH (0.50 g, 2.03 mmol) and compound 235 (1.65 g, 2.22 mmol). The reaction mixture was stirred at 0 °C for 0.5 h, followed by stirring at room temperature for 4 h. The reaction mixture was then concentrated and purified by silica gel column chromatography (acetonitrile / water, 0.1% formic acid 95:5) to give the title compound (1.04 g, 63% yield). ESI-MS m / z [M + H] +:C 32H 56N 5O 17S Calculated value 814.33; Found value 814.46.

[0266] Example 121. Synthesis of compound 237. [] A mixture of compound 236 (0.70 g, 0.86 mmol), N-hydroxysuccinimide (0.20 g, 1.73 mmol), and EDC·hydrochloric acid (1.21 g, 6.36 mmol) in THF (20 mL) was stirred overnight, concentrated under vacuum, and purified on a silica gel column. The purified compound was dissociated with ethyl acetate / dichloromethane (1:4 to 2:1), concentrated, and dried under vacuum to give the title compound (0.540 g, 69% yield). ESI-MS m / z [M + H] +: C 36H 59N 6O 19S, calculated 911.34; found 911.42.

[0267] Example 122. Synthesis of compound 238. Compound 237 (36 mg, 0.04 mmol) was dissolved in DMF (5 mL) at 0 °C, and compound 28 (56.8 mg, 0.08 mmol) and N,N-diisopropylethylamine (0.020 mL, 0.12 mmol) were added. The reaction mixture was heated to room temperature and stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 238 (48 mg, 80% yield). ESI-MS m / z: M +C 71H 99FN 11O 22S Calculated value 1508.67; Found value 1508.86.

[0268] Example 123. Synthesis of methyl 4-(bis(2-(acetylthio)ethyl)amino)-4-sideoxybutyrate (240). At 0 °C, thioacetic acid (10 mL, 134 mmol) was added to a DMA (350 mL) solution of methyl 4-(bis(2-((((methylsulfony)oxy)ethyl)amino)-4-sideoxybutyrate (freshly prepared, 90% purity, 8.5 g, approx. 20 mmol), followed by triethylamine (30 mL, 215 mmol). The mixture was then stirred overnight at room temperature, concentrated, diluted with ethyl acetate (350 mL), and washed with saturated sodium bicarbonate (300 mL), brine (300 mL), and 1 M sodium dihydrogen phosphate (300 mL). The organic layer was dried over sodium sulfate, filtered, concentrated, and purified on a silica column, dissociated with ethyl acetate / n-hexane (10%–25% ethyl acetate) to give the title compound (5.1 g, 76% yield). ESI-MS m / z [M + Na] +: C 13H 21NO 5S 2. Calculated value: 358.1; Measured value: 358.2.

[0269] Example 124. Synthesis of 4-(bis(2-(pyridin-2-yldisulfonyl)ethyl)amino)-4-sideoxybutyric acid (241). A solution of sodium hydroxide (5.0 g, 125 mmol) in water (100 mL) was added to a THF (150 mL) solution of methyl 4-(bis(2-(acetylthioethyl)amino)amino)-4-sideoxybutyrate (5.0 g, 14.9 mmol). The mixture was stirred at room temperature for 35 minutes and neutralized to pH 7 with phosphoric acid. Then, a THF (100 mL) solution of PySSPy (26.0 g, 118 mmol) was added, and the mixture was stirred for 4 hours. The mixture was concentrated and purified on a silica gel column, and dissociated with methanol / dichloromethane / acetic acid (1:20 / 0.2) to give the title compound (5.8 g, 85.6% yield). ESI-MS m / z [M + Na] +: C 18H 21N 3O 3S 4 Calculated value 478.0; Found value 478.2.

[0270] Example 125.2 Synthesis of 5-dioxapyrrolidine-1-yl 4-(bis(2-(pyridin-2-yldithioalkyl)ethyl)amino)-4-sideoxybutyrate (242). To a solution of 4-(bis(2-(2-pyridin-2-yldithioetheryl)ethyl)amino)-4-sideoxybutyric acid (5.2 g, 11.5 mmol) in DMA (100 mL), N-hydroxysuccinimide (1.6 g, 13.9 mmol) and EDC·hydrochloric acid (5.0 g, 26.1 mmol) were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column. Dissociation with ethyl acetate / dichloromethane (5% to 15% ethyl acetate) gave the title compound (5.8 g, 85.6% yield). ESI-MS m / z [M + Na] +: C 22H 24N 4O 5S 4 Calculated value 575.1; Found value 575.2.

[0271] Example 126. Synthesis of N-(4-((S)-2-(4-(bis(2-(pyridin-2-yldisulfonyl)ethyl)amino)-4-sideoxybutyramine)propionamine)benzyl)-1-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-sideoxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethylhexahydropyridine-4-azaium (243). Compound 242 (23 mg, 0.04 mmol) was dissolved in DMA (5 mL) at 0 °C, and compound 28 (56.8 mg, 0.08 mmol) and N,N-diisopropylethylamine (0.020 mL, 0.12 mmol) were added. The reaction mixture was heated to room temperature, stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 273 (39 mg, 85% yield). ESI-MS m / z: M +C 57H 65FN 9O 8S 4: Calculated value 1150.38; Found value 1150.45.

[0272] Example 127. Synthesis of 4-(2-pyridyldithio)-4-methylpentanoic acid (245). A mixture of 2,2'-dithiodipyridine (30.0 g, 136.2 mmol) in methanol (80 mL) and 100 mM sodium phosphate buffer (pH 7.5, 70 mL) was added to a methanol (15 mL) solution of 4-mercapto-4-methylpentanoic acid (Goff, D. et al., BioConjugate Chem. 1990, 1, 381-386) (4.67 g, 31.5 mmol). After stirring for 6 hours, the mixture was concentrated and extracted with ethyl acetate / n-hexane (1:1). The aqueous solution was adjusted to pH 3 and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over sodium sulfate, filtered, evaporated, and purified on a silica gel column (methanol / dichloromethane / acetic acid, 1:15:0.01) to give the title compound (7.05 g, 87%). ESI-MS m / z: [M + H] +C 11H 15NO 2S 2 Calculated value 258.05; Measured value 258.05.

[0273] Example 128. Synthesis of N-succinimino-4-(2-pyridyldithio)-4-methylvalerate (246). To a solution of 4-(2-pyridyldithio)-4-methylpentanoic acid (2.0 g, 7.78 mmol) in dichloromethane (20 mL), N-hydroxysuccinimide (1.10 g, 9.56 mmol) and EDC hydrochloric acid (4.0 g, 20.8 mmol) were added. The mixture was stirred overnight, concentrated, and purified on a silica gel column (ethyl acetate / dichloromethane, 1:10) to give the title compound (2.48 g, 90%). ESI-MS m / z: [M + Na] + C 15H 18N 2O 4S 2 Calculated value 377.07; Found value 377.08.

[0274] Example 129. Synthesis of ((S)-4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3,14-dioxy-3,4,12,14-tetrahydro-1H-guanano[3',4':6,7]indolano[1,2-b]quinoline-11-yl)methyl)-N,N-dimethyl-N-(4-((S)-2-(4-methyl-4-(phenyldisulfonyl)pentanyl)propionic)benzyl)hexahydropyridine-4-azaium (247). Compound 246 (15 mg, 0.04 mmol) was dissolved in DMA (5 mL) at 0 °C, and compound 28 (56.8 mg, 0.08 mmol) and N,N-diisopropylethylamine (0.020 mL, 0.12 mmol) were added. The reaction mixture was heated to room temperature, stirred for 2 hours, concentrated, and purified by preparative HPLC (acetonitrile / water containing formic acid) to give compound 247 (32 mg, 86% yield). ESI-MS m / z: M +C 51H 60FN 6O 7S 2: Calculated value 951.39; Found value 951.39.

[0275] Example 130. Synthesis of (...

Claims

1. A cell surface-binding molecule-camptothecin analog conjugate having the structure shown in formula (I): (I) a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polymorphic crystal structure of such compounds; or an isotope, optical isomer, racemate, non-mirror image isomer, or mirror image isomer thereof; wherein T is a targeting or binding ligand; L is a releasable linker; is a linker bond that independently connects L to an atom of R1, R2, R3, or R5 within parentheses; n is 1-30, m is 1-10; and the parentheses contain an active camptothecin analog, wherein: R1 and R2 are independently H; straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy, alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently H, C(O)NH, C(O)O, SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, C(O)R6. C(O)NHR6; C1-C6 straight-chain or branched alkyl, alkyl alcohol, alkylamine (including first, second, and third amines or quaternary ammonium) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl acetylamine, oxime; C5~C12 glycoside, NH2 or OH; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NHR6, N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is H, C(O)O, C(O)NH, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; or a pharmaceutically acceptable salt; Furthermore, R1, R2, R3 and R6 may be omitted independently, and R2, R3, X, C-10 and C-9 may be used together to form 5-, 6- Or a 7-membered heterocyclic ring;Cell surface binding agents / molecules T are selected from the following groups of components: antibodies; single-chain antibodies; antibody fragments that bind to target cells; monoclonal antibodies; single-chain monoclonal antibodies; monoclonal antibody fragments that bind to target cells; chimeric antibodies; chimeric antibody fragments that bind to target cells; domain antibodies; domain antibody fragments that bind to target cells; adnectin antibodies; DARPins; lymphocytes; hormones; vitamins; growth factors; colony-stimulating factors; nutrient transport molecules (transferrin), and / or peptides, proteins, antibodies, or small molecules bound to albumin, polymers, dendritic macromolecules, liposomes, nanoparticles, vesicles, or (viral) capsids; L has the following structure: , where: -W- is an extension unit; w is 0 or 1; each -Aa- is independently an amino acid unit; r is independently an integer from 0 to 12; -V- is a spacer unit; v is 0, 1, or 2; When the extended body (-W-) is present, it acts as a linker between the cell-binding molecule (T) and the amino acid unit (-Aa-) or spacer V. When Aa is absent, examples of W linked to T are shown below: ; ... -C8 alkyl-(C1-C8 carbon ring)-, -(C3-C7 carbon ring)-C1-C9 alkyl-, -C3-C8 heterocyclic group, -C1-C8 alkyl-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C9 alkyl-, -(CH2CH2O)k-, -(CH(CH3)CH2O)k- and -(CH2CH2O)k-CH2-; k is an integer from 1 to 20; R' and R'' are independently H or CH3; -(Aa)r can be a natural or non-natural amino acid, and can be a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide unit with the same or different amino acid sequences, where r is an integer between 0 and 12; the insulator (-V-) is self-destructive or non-self-destructive; self-destructive includes aromatic compounds of p-aminobenzylaminomethyl (PAB), 2-aminoimidazol-5-methanol derivatives, heterocyclic PAB derivatives, β-glucuronic acid, oro- or p-aminobenzyl acetals, or one of the following structures: , , , , ; (*) marked atoms are insulators or releasable linker units, amino acid (Aa)r, camptothecin analogs, and / or linker sites of cell-binding molecules T; X, Y, and Z3 are independently NH, O, or S; Z2 is independently H,NH, O, or S; v is 0 or 1; Q is independently H, OH, C1-C6 alkyl, (OCH2CH2)n, F, Cl, Br, I, OR17 or SR17, NR17R18, N=NR17, N=R17, NR17R18, NO2, SOR17R18, SO2R17, SO3R17, OSO3R17, PR17R18, POR17R18, PO2R17R18, OPO(OR17)(OR18) or OCH2PO(OR17)(OR18), wherein R17 and R18 are independently H, C1-C8 alkyl, C2-C8 olefin, alkynyl, heteroalkyl, C3-C8 aryl, heterocyclic, carbocyclic, cycloalkyl, heterocyclic, heteroaryl, alkylcarbonyl, or a pharmaceutically acceptable positive ionic salt; v It is an integer from 1 to 20; Examples of non-self-destructive intervals (-V-): ; ... Maleiminoethyl (ME), 4-thio-2-hydroxysulfonyl-butyryl (2-Sulfo-SPDB), aryl thiol (PySS), (4-acetyl)aminobenzoyl (SIAB), oxybenzyl thio, aminobenzyl thio, dioxybenzyl thio, diaminobenzyl thio, aminooxybenzyl thioether, alkoxyamino (AOA), ethyleneoxy (EO), 4-methyl-4-dithiovalerate (MPDP), triazole, dithio, alkylsulfonylalkylsulfonylamine, sulfonylbisamine, phosphine diamine, alkylphosphine diamine, hypophosphinoic acid, N-methylphosphine, N,N'-dimethylphosphine, N,N'-dimethylphosphine diamine, hydrazine, acetamiprid; oxime, diacetylhydrazine, aminoethylamine, aminoethyl- Aminoethylamine, or L- or D-, or a natural or non-natural peptide containing 1-20 amino acids; wherein "*" and "" are spacers or releasable linkers, camptothecin analogs, and / or binding sites; m is 1-10; n is 1-20; X2, X3, X4, X5, or X6 are independently selected from the group consisting of NH, NHNH, N(R12), N(R12)N(R12'), O, S,C1-C6 alkyl, C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; CH2OR12, CH2SR12, CH2NHR12 or 1-8 amino acids; wherein R12 and R12' are independently H, C1-C8 alkyl; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, ester, ether or amide with 1-8 carbon atoms; or a polyoxyethylene unit of the formula (OCH2CH2)p or (OCH2CH(CH3))p, wherein p is an integer from 0 to about 1000; L has one of the following structures: ‑(CR15R16)m(Aa)r(CR17R18)n(OCH2CH2)t-, ‑(CR15R16)m(CR17R18)n(Aa)r(OCH2CH2)t-, ‑(Aa)r-(CR15R16)m(CR17R18)n(OCH2CH2)t-, -(CR15R16)m(CR17R18)n(OCH2CH2)r(Aa)t-, -(CR15R16)m-(CR17=CR18)(CR19R20)n(Aa)t(OCH2CH2)r-, -(CR15R16)m(NR11CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m(Aa)t(NR21CO)(CR19R20)n(OCH2CH2)r-, -(CR15R16)m(OCO)(Aa)t(CR19R20)n(OCH2-CH2)r-, -(CR15R16)m(OCNR17)(Aa)t(CR19R20)n(OCH2CH2)r-, -(CR15R16)m-(CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m(NR21CO)(Aa)t(CR19R20)n(OCH2CH2)r-, -(CR15R16)m(OCO)(Aa)t-(CR19R20)n-(OCH2CH2)r-, -(CR15R16)m(OCNR17)(Aa)t(CR19R20)n-(OCH2CH2)r-、 -(CR15R16)m-(CO)(Aa)t(CR19R20)n-(OCH2CH2)r-、 -(CR15R16)m-phenyl-CO(Aa)t-(CR17R18)n-、 -(CR15R16)m-furan-CO(Aa)t(CR17R18)n-、 -(CR15R6)m-oxazol-CO(Aa)t(CR17R18)n-、-(CR15R16)mthiazole-CO-(Aa)t(CCR17R18)n-, -(CR15R16)t-thiophene-CO(CR17R18)n-, -(CR15R16)t-imidazol-CO-(CR17R18)n-, -(CR15R16)t-morpholine-CO(Aa)t(CR17R18)n-, -(CR15R16)t-piperazine-CO(Aa)t-(CR17R18)n-, -(CR15R16)tN-methylpiperazine-CO(Aa)t(CR17R18)n-, -(CR15R16)m-(Aa)tphenyl-, -(CR15R16)m-(Aa)tfuran-, -(CR15R16)m-oxazole(Aa)t- -(CR15R16)m-thiazole(Aa)t-, -(CR15R16)m-thiophene-(Aa)t-, -(CR15R16)m-imidazol(Aa)t-, -(CR15R16)m-morpholine-(Aa)t-, -(CR15R16)m-piperazine-(Aa)t-, -(CR15R16)mN-methylpiperazine-(Aa)t-, -K(CR15R16)m(Aa)r(CR17R18)n-(OCH2CH2)t-, -K(CR15R16)m-(CR17R18)n(Aa)r(OCH2CH2)t-, -K(Aa)r(CR15R16)m(CR17R18)n-(OCH2CH2)t-, ‑K(CR15R16)m-(CR17R18)n(OCH2CH2)r(Aa)t-、 ‑K(CR15R16)m(CR17=CR18)(CR19R20)n(Aa)t(OCH2CH2)r, ‑K(CR15R16)m(NR11CO)(Aa)t-(CR19R20)n(OCH2CH2)r-, ‑K(CR5R6)m(Aa)t-(NR21CO)(CR19R20)n-(OCH2CH2)r-, ‑K(CR15R16)m(OCO)(Aa)t(CR19R20)n-(OCH2CH2)r-, ‑K(CR15R16)m(OCNR17)(Aa)t-(CR19R20)n-(OCH2CH2)r-, ‑K(CR15R16)m(CO)(Aa)t(CR19R20)n(OCH2-CH2)r-, ‑K(CR15R16)m(NR21-CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, ‑K(CR15R16)m(OCO)(Aa)t-(CR19R20)n(OCH2CH2)r-,-K-(CR15R16)m-(OCNR17)(Aa)t-(CR19R20)n(OCH2CH2)r-, -K(CR15R16)m-(CO)(Aa)t(CR19R20)n-(OCH2CH2)r-, -K(CR15R16)m-phenyl-CO(Aa)t(CR17R18)n-, -K-(CR15R16)m-furan-CO(Aa)t(CR17R18)n, -K(CR15R16)m-oxazol-CO(Aa)t(CR17R18)n-, -K(CR15R16)m-thiazole-CO(Aa)t(CR17R18)n, -K(CR15R16)t-thiophene-CO(CR17R18)n-, -K(CR15R16)timidazol-CO-(CR17R18)n, -K(CR5R6)tmorpholine-CO(Aa)t-(CR17R18)n-, -K(CR15R16)t-piperazine-CO(Aa)t-(CR17R18)n-, -K(CR15R16)tN-methyl-piperazine-CO(Aa)t(CR17R18)n-, -K(CR15R16)m(Aa)t-phenyl, -K-(CR15R16)m(Aa)tfuran-, -K(CR15R16)m-oxazol-(Aa)t-, -K(CR15R16)mthiazole(Aa)t, -K(CR15R16)mthiophene-(Aa)t-, -K(CR15R16)m-imidazolium(Aa)t-, -K(CR15R16)m-morpholine(Aa)t, -K(CR15R16)m-piperazine(Aa)tG, -K(CR5R6)mN-methyl-piperazine(Aa)t-; where Aa, m, n are defined above; t and r are independently 0–100; R13, R14, R15, R16, R17, R18, R19, R20, and R21 are independently selected from group H of the following components: halogens; C1~C8 alkyl or heteroalkyl, C2~C8 aryl, alkenyl, alkynyl, ether, ester, amine or amide, C3~C8 aryl, these groups can be replaced by one or more halogens, CN, NR12R12', CF3, OR12, Aryl, heterocyclic, S(O)R12, SO2R12, -CO2H, -SO3H, -OR12, -CO2R12, -CONR12, -PO2R12R13, -PO3H or P(O)R12R12'R13; K is NH, NR12, -SS-, -C(=O)-, -C(=O)NH-, -C(=O)O-, -C=NH-O-, -C=N-NH-,-C(=O)NH-NH-, O, S, Se, B, Het (heterocyclic or heteroaromatic rings with C3-C12); or peptides containing 1-20 amino acids, either the same or different.

2. The camptothecin analog conjugate of claim 1, having a structure as shown in formula (IV): (IV) or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of such compounds; or isomers, optical isomers, racemic mixtures, non-mirror image isomers, or mirror image isomers thereof; wherein T is a targeting or binding ligand; L is a releasable linker; n is 1-30 and m is 1-10; (in parentheses) is an effective camptothecin analog, wherein: R1 and R2 are independently H, NR6R6', -N+R6R6'R6'', OH, SH, C1-C6 straight-chain or branched alkyl, alkoxy, alkylamino (including first, second, third amino or quaternary ammonium), oxycarbonyl, aminocarbonyl, aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, oxycycloalkyl, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkyl ether, aminoalkyl ether, oxyalkyl ester, aminoalkyl ester, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently -NHC(O)-, -C(O)-, SO2-, -SO2NH-, -NR6SO2-, R6NHC(O), R6OC(O), SO2R6, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, R6C(O), C(O)N R6R6'; Straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including primary, secondary, and tertiary amines) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl acetylamine, oxime; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NH(R6)S(O2)R6', N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is H, C(O)OR6, C(O)NHR6, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkylamide or amino acid; or pharmaceutical salt;Furthermore, R3 is independently defaulted, and X of C-10 is directly connected to L, and R2, R3, X, C-10, and C-9 can collectively form 5-, 6-, or 7-membered heterocycles.

3. The camptothecin analog conjugate of claim 1, having a structure as shown in formula (V): (V) or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof; or a polycrystalline structure of such compounds; or isomers, optical isomers, racemic mixtures, non-mirror image isomers, or mirror image isomers thereof; wherein T is a targeting or binding ligand; L is a releasable linker; n is 1-30, m is 1-10; and the parentheses enclose an active camptothecin analog, wherein: R1 and R2 are independently H, NR6R6', -N+R6R6'R6'', OH, SH, straight-chain or branched C1-C6 alkyl, alkoxy, alkylamino (including first, second, third amino or quaternary ammonium), oxycarbonyl, aminocarbonyl, aminoalkyl, oxyalkyl, aminoalkylamino, oxyalkylamino, aminoalkoxy, oxyalkoxy or alkylcarboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, oxycycloalkyl, aminocycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, aminoalkylcarbonyl, oxyalkylcarbonyl, alkyl ether, alkyl ester, alkylamide, oxyalkyl ether, aminoalkyl ether, oxyalkyl ester, aminoalkyl ester, oxyalkylamide, aminoalkylamide, oxime; NH2 or OH; R3 is independently R6NHC(O)-, R6C(O)-, R6SO2, -SO2NHR6. R6OC(O), R6'SO2R6-, SO3R6, PR6R6', POR6R6', CH2OP(O)(OR6)2, C(O)OP(O)(OR6)2, PO(OR6)(OR6'), P(O)(OR6)OP(O)(OR6')2, R6C(O), C(O)N R6R6'; Straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including primary, secondary, and tertiary amines) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl acetylamine, oxime; R4 is a halogen (F, Cl, Br, or I), CN, NO2, SO3H, OR6, SR6, S(O2)R6, NH(R6)S(O2)R6', N(R6)(R6'), C(O)XR6, N+(R6)(R6')(R6''); X is NH or O; R5 is C(O)O, C(O)NH, R6C(O), straight-chain or branched C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium), alkyl carboxylic acid; C2-C6 carbonate, urea, heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; R6, R6', and R6'' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amine or quaternary ammonium) or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkyl carbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; or pharmaceutical salt; in addition, R5 is independently defaulted and the O of C-20 is directly connected to L, and R2, R3, X, C-10 and C-9 can together form 5-, 6- or 7-membered heterocycles.

4. Camptothecin analogue conjugates as claimed in claim 2 or 3, wherein the camptothecin analogue is connected to linker L, having the structures shown as IV-1 to IV-47 and V-1 to V-61: IV-1, IV-2, IV-3, IV-4, IV-5, IV-6, IV-7, IV-8, IV-9, IV-10, IV-11, IV-12, IV-13, IV-14, IV-15, IV-16, IV-17, IV-18, IV-19, IV-20, IV-21, IV-22, IV-23, IV-24, IV-25, IV-26, IV-27, IV-28, IV-29, IV-30, IV-31, IV-32, IV-33, IV-34, IV-35, IV-36, IV-37, IV-38, IV-39, IV-40. IV-41, IV-42, IV-43, IV-44, IV-45, IV-46, IV-47, V-01, V-02, V-03, V-04, V-05, V-06, V-07, V-08, V-09, V-10, V-11, V-12, V-13, V-14, V-15, V-16, V-17, V-18, V-19, V-20, V-21, V-22, V-23, V-24, V-25, V-26, V-27, V-28, V-29, V-30, V-31, V-32, V-33, V-34, V-35, V-36, V-37, V-38, V-39, V-40, V-41, V-42, V-43, V-44, V-45, V-46, V-47, V-48, V-49, V-50, V-51, V-52, V-53, V-54, V-55, V-56, V-57, V-58, V-59, V-60, V-61, or pharmaceutically acceptable salts, hydrates, or hydrated salts thereof; or polycrystalline structures of such compounds; or isomers, optical isomers, racemates, non-mirror image isomers, or mirror image isomers thereof; wherein "" is a site connected to linker L; wherein R6, and R6' are independently H, C1-C6 alkyl, alkyl alcohol, alkylamine (including first, second, third amines, or quaternary ammonium), or alkyl carboxylic acid; C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, heterocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, alkyl ether, alkyl ester, alkyl amide or amino acid; or pharmaceutical salt.

5. A conjugate of a camptothecin analogue as claimed in claims 1, 2, or 3, wherein the linker L preferably contains an amino group, sulfonamide, phosphatamide, or amino acid group, wherein formula (Iq) can be attached as a branch to the linker L; the amino acid in the linker L is preferably aspartic acid, glutamic acid, lysine, ornithine, or tyrosine, wherein one or both of the functional groups such as amino, carboxyl, or phenoxy can be attached to a long branch of formula (Iq): (Iq); where is the site attached to the sulfonyl, phosphate, amino, or carbonyl group in the linker; G1 is NH, NHNH, C(=O), NHNHC(O), C(=O)NH, C(=NH)NH, CH2, CH2C(O), C(O)O, NHC(O)NH, or (Aa)r, (r = 1-12); G2 is NH, NHNH, C(=O), NHNHC(O) C(=O)NH, C(=NH)NH, CH2, C(O)O, NHC(O)NH, O, S, B, P(O)(OH), NHP(O)(OH), NHP(O)(OH)NH, CH2P(O)(OH)NH, OP(O)(OH)O, CH2P(O)(OH)O, NHS(O)2, NHS(O)2NH, CH2S(O)2NH, OS(O)2O, CH2S(O)2O, Ar, ArCH2, ArO, ArNH, ArS, ArNR1, (Aa)r, (r = 1-12); X1 and X2 are independently O, CH2, S, NH, N(R12), +NH(R12), +N(R12)(R13), C(O), OC(O), OC(O)O, NHSO2NH, NHP(O)(NH)2, SO2NH, P(O)(NH)2, NHS(O)NH, NHP(O)(OH)(NH), OC(O)NH, NHC(O)NH; Y2 is O, NH, NR1, CH2, S, Ar;G3 is OH, SH, OR1, SR1, OC(O)R1, NHC(O)R12, C(O)R12, CH3, NH2, NR12, +NH(R12), +N(R12)(R13), C(O)OH, C(O)NH2, NHC(O)NH2, BH2, BR12R13, P(O)(OH)2, NHP(O)(OH)2, NHP(O)(NH2)2, S(O)2(OH), (CH2)q1C(O)OH, (CH2)q1P(O)(OH)2, C(O)(CH2)q1C(O)OH, OC(O)(CH2)q1C(O)OH, NHC(O)(CH2)q1C(O)OH, CO(CH2)q1P(O)(OH)2, NHC(O)O(CH2)q1-C(O)OH, OC(O)NH-(CH2)q1C(O)OH, NHCO(CH2)q1P(O)(OH)2, NHC(O)(NH)(CH2)q1C(O)OH, CONH(CH2)q1-P(O)(OH)2, NHS(O)2(CH2)q1C(O)OH, CO(CH2)q1S(O)2(OH), NHS(O)2NH-(CH2)q1C(O)OH, OS(O)2NH(CH2)q1C(O)OH, NHCO(CH2)q1S(O)2(OH), NHP(O)(OH)(NH)-(CH2)q1C(O)OH, CONH(CH2)q1S(O)(OH), OP(O)(OH)2, (CH2)q1P(O)(NH)2, NHS(O)₂(OH), NHS(O)₂NH₂, CH₂S(O)₂NH₂, OS(O)₂OH, OS(O)₂OR₁, CH₂S(O)₂OR₁, Ar, ArR₁₂, ArOH, ArNH₂, ArSH, ArNHR₁₂, or (Aa)q₁; p₁, p₂, and p₃ are independently 0 -30 but not simultaneously 0; q₁ and q₂ are independently 0 -24;Preferably, G3 is a linear or branched C2-C50 polycarboxylic acid or C2-C50 polyalkylamine, C6-C50 oligosaccharide or polysaccharide, C6-C50 betaine zwitterion or poly(sulfobetaine) zwitterion (PSBs) containing quaternary ammonium cations and sulfonate anions, biodegradable polymers such as polylactic acid / glycolic acid (PLGA), poly(acrylate), deacetylated chitosan, copolymers of N-(2-hydroxypropyl)methacrylamide, poly[2-(methacryloxy)ethylphosphocholine] (PMPC), poly-L-glutamic acid, poly(lactide-co-glycolic acid) (PLG), poly(ethylene glycol) (PEG), or poly(propylene glycol) (PPG). Poly(ethylene glycol) modified peptides, poly(ethylene glycol) modified liposomes, poly(ethylene glycol) modified alkyl carboxylic acids, poly(ethylene glycol) modified alkylamines, hyaluronic acid (HA) (glycosaminoglycans), heparin / acetylheparin sulfate (HSGAG), chondroitin sulfate / corticotin sulfate (CSGAG), poly(ethylene glycol) modified alkyl sulfates, poly(ethylene glycol) modified alkyl phosphates, or poly(ethylene glycol) modified alkyl quaternary ammonium salts.

6. A conjugate of camptothecin analogues as claimed in claim 5, wherein the branched linkers of formula (Iq) are selected from the group consisting of: (I-q01), (I-q02), (I-q03), (I-q04), (I-q04), (Iq-05), (I-q06), (I-q07), (I-q08), (Iq-09), (Iq-10), (Iq-11), (Iq-12), (Iq-13), (Iq-14), wherein G1, p1, p2, p3, Aa, r, X2, q1, and m1 are the same as defined in claim 7.

7. Conjugates of camptothecin analogues as claimed in claims 1, 2, or 3, wherein the camptothecin analogue conjugates of formulas (I), (IV), and (V) can be readily conjugated by cell-binding molecule T with compounds as shown in formulas (VI), (IX), and (X):

1. (VI) (IX) (X) where R1, R2, R3, R4, R5, L, X, and m are as defined in formula (I); Lv is a reactive functional group that can react with thiols, amines, carboxylic acids, selenools, phenols, or hydroxyl groups on cellular molecules. It is selected from the following group of halogens (fluorine, chlorine, bromine, iodine): maleimino; methanesulfonyl; toluenesulfonyl; trifluoromethanesulfonyl; nitrophenoxy; N-hydroxysuccinimino (NHS); phenoxy; dinitrophenoxy; pentafluorophenoxy; tetrafluorophenoxy; trifluorophenoxy; difluorophenoxy; monofluorophenoxy; pentachlorophenoxy; imidazole; monochlorobenzene. Oxygen group; dichlorophenoxy; trichlorophenoxy; tetrachlorophenoxy; N-(benzotriazolyl)oxy; 2-ethyl-5-phenylisooxazolyl-3′-sulfonyl; phenoxydiazolylsulfonyl; 2-ethyl-5-phenylisooxazolyl; phenoxydiazolyl; oxadiazolyl; unsaturated carbon (double or triple bonds between carbon-carbon, carbon-nitrogen, carbon-sulfur, carbon-phosphorus, sulfur-nitrogen, phosphorus-nitrogen, oxygen-nitrogen, or carbon-oxygen); or intermediates formed by reaction with Mitsunobu's reagent; examples of condensing agents are as follows: 1-Ethyl-(3-dimethylaminopropyl)carbodiimide (EDC), dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), N-cyclohexyl-N'-(2-morpholino-ethyl)carbodiimide methyl p-toluenesulfonate (CMC or CME-CDI), 1,1'-carbonyldiimidazole (CDI), (O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TBTU), N,N,N',N'-tetramethyl-oxo-(1H-benzotriazol-1-yl)-ammonium hexafluorophosphate (HBTU), ( Benztriazol-1-yloxy)tris(dimethylamino)-hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinylhexafluorophosphate (PyBOP), diethyl cyanophosphonate (DEPC), chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine-1-oxohexafluorophosphate (HATU), 1-[(dimethylamino)(morpholino)methylene]-1H-[1,2,3]triazolo[4,5-b]pyridine-1-onthium-3-oxohexafluorophosphate (HDMA), 2-chloro-1,3-dimethyl-imidazolium hexafluorophosphate (CIP), chloropyrrolidone-onthium hexafluorophosphate (PyCloP), fluoro- N,N,N',N'-bis(tetramethylene)formamidinium hexafluorophosphate (BTFFH), N,N,N',N'-Tetramethyl-S-(1-sideoxy-2-pyridyl)thiourea hexafluorophosphate, O-(2-sideoxy-1(2H)pyridyl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TPTU), S-(1-sideoxy-2-pyridyl)N,N,N',N'-tetramethylthiourea tetrafluoroborate, O-[(ethoxycarbonyl)-cyanomethylamino]-N,N,N',N'-tetramethylurea hexafluorophosphate (HOTU), (1-cyano-2-ethoxy-2-sideoxyethylaminooxy)dimethylamino-morpholino-hexafluorophosphate (COMU), O-(benzotriazol-1-yl)- N,N,N',N'-bis(tetramethylene)hexafluorophosphate (HBPyU), N-benzyl-N'-cyclohexyl-carbodiimide (with or without polymer binding), dipyrrolidinyl(N-succiniminooxy)carbomonyhexafluorophosphate (HSPyU), dichloropyrrolidinylhexafluorophosphate (PyClU), 2-chloro-1,3-dimethylimidazolium tetrafluoroborate (CIB), (benzotriazol-1-yloxy)diguanidine carbomonyhexafluorophosphate (HBPipU), O-(6-chlorobenzotriazol-1-yl)- N,N,N',N'-Tetramethylurea tetrafluoroborate (TCTU), brominated (dimethylamino)-hexafluorophosphate (BroP), propylphosphonic anhydride (PPACA, T3P), 2-morpholinoethyl isocyanate (MEI), N,N,N',N'-tetramethyl-oxo-(N-succinimino)hexafluorophosphate (HSTU), 2-bromo-1-ethyl-pyridinium tetrafluoroborate (BEP), O-[(ethoxycarbonyl)cyano-methyleneamino]-N,N,N',N'-tetramethylurea tetrafluoroborate (TOTU), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine chloride (MMTM, DMTMM), N,N,N',N'-tetramethyl-oxo-(N-succinimino) Urea tetrafluoroborate (TSTU), O-(3,4-dihydro-4-sideoxy-1,2,3-benzotriazine-3-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TDBTU), 1,1'-(azodicarbonyl)-diguanidine (ADD), di-(4-chlorobenzyl)azodicarboxylate (DCAD), di-tert-butyl azodicarboxylate (DBAD), diisopropyl azodicarboxylate (DIAD), diethyl azodicarboxylate (DEAD); Additionally, Lv1 and Lv2 can be acid anhydrides.Anhydrides formed by the self-condensation of acids or by their reaction with other C1-C8 anhydrides; preferred Lv includes the following substances: halides (such as fluorides, chlorides, bromides, and iodides), mesyl, tosyl, trifluoromethylsulfonyl, trifluoromethylsulfonate, nitrophenoxy, N-succinimino (NHS), phenoxy; dinitrophenoxy; pentafluorophenoxy, tetrafluorophenoxy, trifluorophenoxy, difluorophenoxy, monofluorophenoxy, pentachlorophenoxy, 1H-imidazol-1-yl, monochlorophenoxy, dichlorophenoxy, trichlorophenoxy, tetrachlorophenoxy, N-(benzotriazolyl)oxy, 2-ethyl-5-phenylisoxazole-3′- Sulfoyl, phenyloxadiazole-sulfoyl, 2-ethyl-5-phenylisooxazolyl, phenyloxadiazole, oxadiazole, unsaturated carbon (double or triple bonds between carbon-carbon, carbon-nitrogen, carbon-sulfur, carbon-phosphorus, sulfur-nitrogen, phosphorus-nitrogen, oxygen-nitrogen, or carbon-oxygen), or one of the following structures: disulfide; haloacetyl; acetyl halide; N-hydroxysuccinimide; maleimide; monosubstituted maleimide; disubstituted maleimide; monosubstituted succinimide; disubstituted maleimide; substituted maleic acid; -CHO Aldehydes; ethylenesulfonyl; acrylyl; 2-(toluenesulfonoxy)acetyl; 2-(methanesulfonoxy)acetyl; 2-(nitrophenoxy)acetyl; 2-(dinitrophenyl)acetyl; 2-(fluorophenoxy)acetyl; 2-(difluorophenoxy)acetyl; 2-((trifluoromethylsulfonyl)oxy)acetyl; ketones or aldehydes, 2-(pentafluorophenoxy)acetyl; methyl benzophenyl diazole (ODA); acid anhydrides, alkoxyamines; azide, alkynyl, or acehydrazine; wherein, X1' is F, Cl, Br, I, or Lv3; X2' is O, NH, N(R1), or CH2; R3 is H, aryl, or heteroaryl, wherein one or more H atoms may be independently substituted by -R1, -halogen, -OR1, -SR1, -NR1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1, wherein R1 and R2 are defined above; Lv3 is a leaving group selected from the group consisting of F, Cl, Br, I; nitrophenoxy; N-hydroxysuccinimide (NHS); phenoxy; dinitrophenoxy; pentafluorophenoxy; tetrafluorophenoxy; difluorophenoxy; monofluorophenoxy; pentachlorophenoxy; trifluoromethanesulfonyl; imidazolyl; dichlorophenoxy; tetrachlorophenoxy; 1-Hydroxybenzotriazolyl; toluenemethanesulfonyl; methanesulfonyl; 2-ethyl-5-phenylisoxazole-3′-sulfonyl, anhydride or anhydride formed by the reaction of anhydride with other anhydrides.

8. A conjugate of a camptothecin analogue as claimed in claim 7, wherein the compounds of formulas (VI), (IX) and (X) that can be used to prepare CPT conjugates have the following compound structures: 56, 61, 70, 75, 81 and 86: and.

9. A conjugate of a camptothecin analogue as claimed in claims 1, 2 or 3, wherein the cell-binding molecule (T or mAb) is selected from the group consisting of: (A): antibodies, proteins, proantibodies, nanobodies, vitamins (including folic acid), peptides, macromolecular micelles, liposomes, lipoprotein-based drugs, nanoparticle drugs, dendritic polymers, and the above molecules or particles coated on or linked to cell-binding ligands, or combinations thereof; (B): Antibody-like proteins, full-length antibodies (polyclonal antibodies, monoclonal antibodies, antibody dimers, antibody multimers), multispecific antibodies (bispecific antibodies, trispecific antibodies, or tetraspecific antibodies), single-chain antibodies, antibody fragments that bind to target cells, monoclonal antibodies, single-chain monoclonal antibodies, monoclonal antibody fragments that bind to target cells, chimeric antibodies, chimeric antibody fragments that bind to the target, domain antibodies, domain antibody fragments that bind to target cells, surface recombinant antibodies, surface recombinant single-chain antibodies, surface recombinant antibody fragments that bind to target cells, humanized antibodies or surface recombinant antibodies, humanized single-chain antibodies, humanized antibody fragments that bind to target cells, anti-idiotype (anti-Id) antibodies, CDR, biantibodies, triantibodies, tetraantibodies, small antibodies, proantibodies, proantibody fragments, small immunoproteins (SIPs), lymphocytes, hormones, vitamins, growth factors, colony-stimulating factors, nutrient transport molecules, high molecular weight proteins, fusion proteins, kinase inhibitors, gene targeting agents, nanoparticles or polymers modified with antibodies or high molecular weight proteins; (C): Cell-binding ligands or receptor agonists selected from the following groups: folic acid derivatives; urea glutamate derivatives; somatostatin and its analogues (selected from the following groups: octreotide (Sandostatin) and somatuline); aromatic sulfonamides; pituitary adenylate cyclase initiating peptide (PACAP) (PAC1); angiotensin-converting peptides (VIP / PACAP) (VPAC1, VPAC2); melanocyte-stimulating hormone (α-MSH); cholecystokinin (CCK) / gastrin receptor and its receptor subtypes; bufotalin (selected from the following groups: Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2) / gastrin-releasing peptide (GRP); neurotensin receptor and its receptor subtypes (NTR1, NTR2, NTR3); substance P (NK1 receptor) ligand; neuropeptide Y (Y1–Y6); homing peptides include RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimer and polycyclic RGD peptides (selected from the group cRGDfV of the following composition).TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor) and F3 peptide; cell-penetrating peptides (CPPs); peptide hormones, agonists and antagonists of luteinizing hormone-releasing hormone (LHRH) selected from the following groups, gonadotropin-releasing hormone (GnRH) agonists, acting by targeting follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and testosterone production, selected from the following groups, busereline (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), and gonadotropin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2). Goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH2), Histaminerelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), Leuprorelin (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), Nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH2), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2), Nafarelin, Abarelin (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropylLys-Pro-DAla-NH2), (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2), Leupron (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(carba-mo yl)-Leu-isopropylLys-Pro-D-Ala-NH2), and degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9, N10-diethyl)-homoArg-Leu-(N9,N10-diethyl)-homoArg-Pro-D-Ala-NH2); Pattern recognition receptors (PRRs), Toll-like receptors (TLRs), C-type lectins and nodular receptors (NLRs) selected from the following groups; Calcitonin receptor agonists; Integrin receptors and their receptor subclasses (selected from the following groups: αVβ1, αVβ3, αVβ5, αVβ6, α6β4, α7β1, αLβ2, αIIbβ3) agonists (selected from the following groups: GRGDSPK, cyclo(RGDfV) (L1) and its derivatives [cyclo(-N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N(Me)fV), cyclo(RGDf-N(Me)V-)(Cilengitide)]; Nanobodies ( VHH derivatives (cameloid Ig); domain antibodies (dAb, derivatives of VH or VL domains); bispecific T cell adapters (BiTE, bispecific biantibody); dual affinity retargeting agents (DART, bispecific biantibody); tetravalent tandem antibodies (TandAb, dimeric bispecific biantibody); anticalin (a derivative of Lipocalins); adnectins (FN3 at position 10 (fibronectin)); designed ankyrin repeats (DARPins); avimers; EGF receptor.Or VEGF receptor agonists; (D): Small molecules of cell-binding molecules / ligands or cell receptor agonists selected from the following groups: LB01 (folic acid), LB02 (PMSA ligand), LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (somatostatin), LB06 (somatostatin), LB07 (octreotide, somatostatin analog), LB08 (lanreotide, somatostatin analog), LB09 (Sanvar, somatostatin analog), LB10 (CAIX ligand), LB11 (CAIX ligand), LB12 (gastrin-releasing peptide receptor (GRPr), MBA), LB13 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligands), LB14 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligands), LB15 ( GnRH antagonist (Abarelix), LB16 (cobalamin, vitamin B12 analog), LB17 (cobalamin, vitamin B12 analog), LB18 (for αvβ3 integrin receptor, cyclic RGD pentapeptide), LB19 (VEGF receptor isodivalent peptide ligand), LB20 (neuromycin B), LB21 (frog dermalin, acting on G protein-coupled receptors), LB22 (TLR2, acting on Toll-like receptors), LB23 (acting on androgen receptors). Sex hormone receptors), LB24 (silengiptide or cyclic (-RGDfV-) conjugate, acting on αv integrin receptors), LB23 (flucotrienol), LB25 (rifabutin analog), LB26 (rifabutin analog), LB27 (rifabutin analog), LB28 (fludrocortisone), LB29 (dexamethasone), LB30 (fluticasone propionate), LB31 (beclomethasone propionate), LB32 (triamcinolone acetonide), LB33 (prednisone), LB34 The following compounds are listed below: Prednisolone, LB35 (methylprednisolone), LB36 (betamethasone), LB37 (irinotecan analog), LB38 (crizotinib analog), LB39 (bortezomib analog), LB40 (carfizzomib analog), LB41 (carfilzomib analog), LB42 (leuprorelin analog), LB43 (triptorelin analog), LB44 (clindamycin), LB45 (liraglutide analog), LB46 (vincristine analog), LB47 (retapalline analog), LB48 (butulin analog), LB49 (vincristine analog), LB50 (lixisentide analog), LB51 (osidinib analog), LB52 (nucleoside analog), LB53 (erlotinib analog), and LB54 (lapatinib analog). Their structures are shown below: LB01 (folic acid).LB02 (PMSA ligand), LB03 (PMSA ligand), LB04 (PMSA ligand), LB05 (Somatostatin), LB06 (Somatostatin), LB07 (Octreotide, Somatostatin analog), LB08 (Lanpeptide, Somatostatin analog), LB09 (Sanvar aminopeptide, Somatostatin analog), LB10 (CAIX ligand), LB11 (CAIX ligand), LB12 (Gastric-releasing peptide receptor (GRPr), MBA), LB13 (LH-RH and Gonadotropin-releasing hormone GnRH ligand), LB14 (LH-RH and Gonadotropin-releasing hormone GnRH ligand), LB15 (GnRH antagonist, Abaric). R19 is 5'-deoxyadenosine, Me, OH, CN; LB16 (cobalamin, vitamin B12 analog), R19 is 5'-deoxyadenosine, Me, OH, CN; LB17 (cobalamin, vitamin B12 analog), LB18 (cyclic RGD pentapeptide, acting on αvβ3 integrin receptor), LB19 (heterodivalent peptide ligand conjugate, acting on vascular endothelial growth factor (VEGF) receptor), LB20 (neuromucoidin B), LB21 (froglin conjugate, acting on G protein-coupled receptor), LB22 (TLR2 conjugate, acting on Toll-like receptor), LB23 (androgen receptor), LB24 (silengitide / cyclic (-RGDfV-) conjugate, acting on αv integrin receptor), LB25 (rifabutin analog), LB26 (rifabutin analog). LB27 (Rifabutin analog), LB28 (Fluordrocortisone), LB29 (Dexamethasone), LB30 (Fluticasone propionate), LB31 (Beclomethasone propionate), LB32 (Triamcinolone), LB33 (Prednisone), LB34 (Prednisolone), LB35 (Methylprednisolone), LB36 (Beflumethasone), LB37 (Irinotecan analog), LB38 (Crizotinib analog), LB39 (Bortezomib analog), where Y5 is N, CH, C(Cl), C(CH3), or C(COOR1); R1 is H, C1-C6 alkyl, or C3-C8 aryl; LB40 (Carfizzomib analog), LB41 (Carfizzomib analog), LB42 (Leuprorelith analog), LB43 (Triptorelin analogue), LB44 (Clindamycin), LB45 (Liraglutide analogue), LB46 (Semaglutide analogue), LB47 (Ritapalene analogue).LB48 (Dublin analog), LB49 (Vincristine analog), LB50 (Lixisentide analog), LB51 (Ositinib analog), LB52 (Nucleoside analog), LB53 (Erlotinib analog), LB54 (Lapatinib analog).

10. A conjugate of a camptothecin analogue as claimed in items 1, 2, or 3, wherein the cell-binding agent / molecule T is effective against the following cells: tumor cells, virus-infected cells, microbial-infected cells, parasite-infected cells, autoimmune cells, activated tumor cells, bone marrow cells, activated T cells, B cells, or melanocytes, or any diseased cells expressing the following antigens or receptors: CD1, CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CD12w, CD13, CD14, CD15, CD16, CD16a, CD16b, CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25. CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD32a, CD32b, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49c, CD49c, CD49d, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CD60, CD60a, CD60b, CD60c, CD61, CD62E, CD62L, CD62P, CD63, CD64, CD65, CD65s, CD66, CD66a, CD66b, CD66c, CD66d, CD66e, CD66f, CD67, CD68, CD69, CD70, CD71, CD72, CD73, CD74, CD75, CD75s, CD76, CD77, CD78, CD79, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, CD85a, CD85b, CD85c, CD85d, CD85e, CD85f, CD85g, CD85g, CD85i, CD85j, CD85k, CD85m, CD86, CD87, CD88,CD89、 CD90、 CD91、CD92、 CD93、 CD94、 CD95、 CD96、 CD97、 CD98、 CD99、 CD100、 CD101、 CD102、 CD103、 CD104、 CD105、 CD106、 CD107、 CD107a、 CD107b、 CD108、 CD109、 CD110、 CD111、 CD112、 CD113、 CD114、 CD115、 CD116、 CD117、 CD118、 CD119、 CD120、 CD120a、 CD120b、 CD121、 CD121a、 CD121b、 CD122、 CD123、 CD123a、 CD124、 CD125、 CD126、 CD127、 CD128、 CD129、 CD130、 CD131、 CD132、 CD133、 CD134、 CD135、 CD136、 CD137、 CD138、 CD139、 CD140、 CD140a、 CD140b、 CD141、 CD142、 CD143、 CD144、 CD145、 CDw145、 CD146、 CD147、 CD148、 CD149、 CD150、 CD151、 CD152、 CD153、 CD154、 CD155、 CD156、 CD156a、 CD156b、 CD156c、 CD156d、 CD157、 CD158、 CD158a、 CD158b1、 CD158b2、 CD158c、 CD158d、 CD158e1、 CD158e2、 CD158f2、 CD158g、 CD158h、 CD158i、 CD158j、 CD158k、 CD159、 CD159a、 CD159b、 CD159c、 CD160、 CD161、 CD162、 CD163、 CD164、 CD165、 CD166、 CD167、 CD167a、 CD167b、 CD168、 CD169、 CD170、 CD171、 CD172、 CD172a、 CD172b、 CD172g、 CD173、 CD174、 CD175、 CD175s、 CD176、 CD177、 CD178、 CD179、 CD179a、 CD179b、 CD180、 CD181、 CD182、 CD183、 CD184、 CD185、 CD186、 CDw186、 CD187、CD188、 CD189、 CD190、 CD191、 CD192、 CD193、 CD194、 CD195、 CD196、 CD197、 CD198、 CD199、 CDw198、 CDw199、 CD200、 CD201、 CD202、 CD202(a、b)、 CD203、 CD203c、 CD204、 CD205、 CD206、 CD207、 CD208、 CD209、 CD210、 CDw210a、 CDw210b、 CD211、 CD212、 CD213、 CD213a1、 CD213a2、 CD214、 CD215、 CD216、 CD217、 CD218、 CD218a、 CD218、 CD21b9、 CD220、 CD221、 CD222、 CD223、 CD224、 CD225、 CD226、 CD227、 CD228、 CD229、 CD230、 CD231、 CD232、 CD233、 CD234、 CD235、 CD235a、 CD235b、 CD236、 CD237、 CD238、 CD239、 CD240、 CD240ce、 CD240d、 CD241、 CD242、 CD243、 CD244、 CD245、 CD246、 CD247、 CD248、 CD249、 CD250、 CD251、 CD252、 CD253、 CD254、CD255、 CD256、 CD257、 CD258、 CD259、 CD260、 CD261、 CD262、 CD263、 CD264、 CD265、 CD266、 CD267、 CD268、 CD269、 CD270、 CD271、 CD272、 CD273、 CD274、 CD275、 CD276、 CD277、 CD278、 CD279、 CD281、 CD282、 CD283、 CD284、 CD285、 CD286、 CD287、 CD288、 CD289、 CD290、 CD291、 CD292、 CD293、 CD294、 CD295、 CD296、 CD297、 CD298、 CD299、 CD300、 CD300a、 CD300b、 CD300c、 CD301、 CD302、 CD303、 CD304、 CD305、 CD306、 CD307、 CD307a、 CD307b、 CD307c、CD307d, CD307e, CD307f, CD308, CD309, CD310, CD311, CD312, CD313, CD314, CD315, CD316, CD317, CD318, CD319, CD320, CD321, CD322, CD323, CD324, CD325, CD326, CD327, CD328, CD329, CD330, CD331, CD332, CD333, CD334, CD335, CD336, CD337, CD338, CD339, CD340, CD341, CD342, CD343, CD344, CD345, CD346, CD347, CD348, CD349, CD350, CD351, CD352, CD353, CD354, CD355, CD356, CD357, CD358, CD359, CD360, CD361, CD362, CD363, CD364, CD365, CD366, CD367, CD368, CD369, CD370, CD371, CD372, CD373, CD374, CD375, CD376, CD377, CD378, CD379, CD381, CD382, CD383, CD384, CD385, CD386, CD387, CD388, CD389, CRIPTO, CRIPTO, CR, CR1, CRGF, CRIPTO, CXCR5, LY64, TDGF1, 4-1BB, APO2, ASLG659, BMPR1B, 4-1BB, 5AC, 5T4 (Trophoblast glycoprotein, TPBG, 5T4, WNT-activating inhibitory factor 1 or WAIF1), adenocarcinoma antigen, AGS-5, AGS-22M6, kappain receptor kinase 1, AFP, AKAP-4, ALK, α-integrin, αvβ6, aminopeptidase N, amyloid-β, androgen receptor, angiogenesis factor 2, angiogenesis factor 3, annexin A1, anthrax toxin protective antigen, anti-transfer protein receptor, AOC3 (VAP-1), B7-H3, Bacillus anthracis, BAFF (B cell activating factor), BCMA, B lymphoma cells, bcr-abl, frog skin extract, BORIS, C5, C242 antigen, CA125 (glycoantigen 125, MUC16), CA-IX (or CAIX, carbonic anhydrase 9),CALLA, CanAg, canine lupus erythematosus IL31, carbonic anhydrase IX, myosin, CCL11 (CC fragment chemokine 11), CCR4 (CC chemokine receptor 4), CCR5, CD3E (ε), CEA (carcinoembryonic antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (factor D), Ch4D5, cholecystokinin 2 (CCK2R), CLDN18 (Claudin-18), CLDN18.2 (Claudin-18.2), clustering factor A, cMet, CRIPTO, FCSF1R (colony-stimulating factor 1 receptor). CSF2 (colony-stimulating factor 2, granulocyte-macrophage colony-stimulating factor (GM-CSF)), CSP4, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), CTAA16.88 tumor antigen, CXCR4, CXC chemokine receptor 4, cyclic ADP ribonuclease, cyclin B1, CYP1B1, cytomegalovirus, cytomegalovirus glycoprotein B, Dabigatran, DLL3 (Δ-ligand 3), DLL4 (Δ-ligand 4), DPP4 (dipeptide-peptidase 4), DR5 (death receptor 5), E. coli shiga toxin type-1, E. coli shiga toxin type-2, ED-B, EGFL7 (EGF-like domain protein 7). EGFR, EGFRII, EGFRvIII, endothelial factor, endothelin B receptor, endotoxin, EpCAM (epithelial cell adhesion molecule), EphA2, Episialin, ERBB2 (epidermal growth factor receptor 2), ERBB3, ERG (TMPRSS2 ETS fusion gene), *Escherichia coli*, ETV6-AML, FAP (fibroblast activation protein α), FCGR1, alpha-fetoprotein, fibrin II β chain, fibronectin extradomain-B, FOLR (folate receptor), folate receptor α, folate hydrolase, Fos-associated antigen 1, respiratory syncytial virus F protein, coiled receptor, fucose GM1, GD2 ganglioside, G-28 (cell surface antigen glycolipid), GD3 idiotype, GloboH, Glypican 3. N-hydroxyacetylneuraminic acid, GM3, GMCSF receptor α chain, growth differentiation factor 8, GP100, GPNMB (transmembrane glycoprotein NMB), GUCY2C (guanylate cyclase 2)C), guanylate cyclase C (GC-C), intestinal guanylate cyclase, guanylate cyclase C receptor, heat-stable enterotoxin receptor (hSTAR), heat shock protein, hemagglutinin, hepatitis B surface antigen, hepatitis B virus, HER1 (human epidermal growth factor receptor 1), HER2, HER2 / neu, HER3 (ERBB-3), IgG4, HGF / SF (hepatocyte growth factor / dispersion factor), HHGFR, HIV-1, histone complex, HLA-DR (human leukocyte antigen), HLA-DR10, HLA-DRB, HMWMAA, human chorionic gonadotropin, HNGF, human dispersion factor receptor kinase, HPV E6 / E7, Hsp90, hTERT, ICAM-1 (intercellular adhesion molecule 1), idiotype, IGF1R (IGF-1, insulin-like growth factor 1 receptor), IGHE, IFN-γ, influenza hemagglutinin, IgE, IgE Fc region, IGE, interleukins (including IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-6R, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-17, IL-17A, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-27, IL-28), IL31RA, ILGF2 (insulin-like growth factor 2), integrins (α4, αIIbβ3, αvβ3, α4β7, α5β1, α6β4, α7β7, αllβ3, α5β5, αvβ5), interferon-γ-induced protein, ITGA2, ITGB2, KIR2D, Kapa Ig, LCK, Le, Legumain, Lewis-Y antigen, LFA-1 (lymphocyte function-associated antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A, LMP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE-2, MAGE-3, MAGE A1, MAGE A3, MAGE 4, MART1, MCP-1, MIF (macrophage migration inhibitory factor, or glycosyl inhibitory factor (GIF)), MS4A1 (transmembrane 4-domain subfamily A member 1), MSLN (mesothelin), MUC1 (mucin 1, cell surface-associated (MUC1) or polymorphic epithelial mucin (PEM)), MUC1-KLH, MUC16 (CA125) MCP1 (monocyte chemoattractant protein 1), MelanA / MART1, ML-IAP, MPG, MS4A1 (transmembrane 4-domain subfamily A)MYCN, myelin-associated glycoprotein, myostatin, NA17, NARP-1, NCA-90 (granulocyte antigen), Nectin-4 (ASG-22ME), NGF, neuronal apoptosis-regulating protease 1, NOGO-A, Notch receptor, nucleolin, Neu oncogene product, NY-BR-1, NY-ESO-1, OX-40, OxLDL (oxidized low-density lipoprotein), OY-TES1, P21, p53 non-mutant, P97, Page4, PAP, anti-(N-hydroxyacetylneuraminic acid) antibody binding site, PAX3, PAX5, PCSK9, PDCD1 (PD-1, programmed cell death protein 1), PDGF-Rα (alpha platelet-derived growth factor receptor), PDGFR-β, PDL-1, PLAC1, PLAP-like testicular alkaline phosphatase, platelet-derived growth factor receptor β, sodium phosphate cotransporter, PMEL 17. Polysialic acid, proteinase 3 (PR1), prostate cancer, PS (phosphatidylserine), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies virus glycoprotein, RHD (Rh polypeptide 1 (RhPI)), Rhesus factor, RANKL, RhoC, Ras mutation, RGS5, ROBO4, respiratory syncytial virus, RON, ROR1, sarcoma translocation interruption point, SART3, Sclerostin, SLAMF7 (SLAM member 7), Selectin P, SDC1 (multi-ligand proteoglycan 1), systemic lupus erythematosus (a), somatostatin C, SIP (sphingosine 1-phosphate), somatostatin, spermin 17, SSX2, STEAP1 (6-transmembrane prostate antigen 1), STEAP2, STn, TAG-72 (tumor-associated glycoprotein 72), susceptin, T cell receptor, T cell transmembrane protein, TEM1 (tumor vascular endothelial marker 1), TENB2, Tenascin C (TN-C), TGF-α, TGF-β (transforming growth factor β), TGF-β1, TGF-β2 (transforming growth factor 2), Tie (CD202b), Tie2, TIM-1 (CDX-014), Tn, TNF, TNF-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor superfamily member 10) B), TNFRSF13B (tumor necrosis factor receptor superfamily member 13B), TPBG (trophoblast glycoprotein), TRAIL-R1 (TNF-associated necrosis-inducing ligand receptor 1), TRAILR2 (death receptor 5 (DR5)), tumor-associated calcium signaling sensor 2, tumor-specific glycosylated MUC1, TWEAK receptor, TYRP1 (glycoprotein 75), TRP-1 (Trop1), TRP-2(Trop2), tyrosinase, VCAM-1, VEGF, VEGF-A, VEGF-2, VEGFR-1, VEGFR-2, vimentin, WT1, XAGE 1, cells expressing insulin growth factor receptor, or cells expressing epidermal growth factor receptor.

11. A conjugate of a camptothecin analogue as claimed in claim 10, wherein the tumor cells are selected from the group of cells comprising: lymphoma cells, myeloma cells, kidney cells, breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cell carcinoma cells, small cell lung cancer cells, non-small cell lung cancer cells, testicular cancer cells, malignant cells, or any cells that cause cancer by uncontrolled accelerated growth and division.

12. A pharmaceutical composition comprising a therapeutically effective amount of a conjugate of a camptothecin analogue of claim 1, 2 or 3, and a pharmaceutically acceptable salt, carrier, diluent or excipient, or a combination thereof, for the treatment or prevention of cancer, autoimmune diseases or infectious diseases.

13. A conjugate of a camptothecin analogue, such as those in claims 1, 2, or 3, having in vitro, in vivo, or ex vivo cytotoxic activity.

14. The pharmaceutical composition of claim 12, characterized in that it is administered simultaneously with a chemotherapeutic agent, radiotherapy, immunotherapy agent, autoimmune dysregulator, anti-infective agent or other conjugate to synergistically treat or prevent cancer, autoimmune diseases or infectious diseases.

15. The pharmaceutical composition of claim 14, wherein the chemotherapeutic agent is selected from any one or more of the following groups: 1) a) Alkylating agent: nitrogen mustard selected from the following groups: chlorpheniramine, cloprazosin, cyclophosphamide, dacarbazine, estradiol nitrogen mustard, ifosfamide, nitrogen mustard, dimethoxyamine hydrochloride, dioxane mustard oxide, amlodipine hydrochloride, mycophenolic acid, eugenol, guanobromide, neonitrogen mustard, benzyl mustard cholesterol, pine oxychloride, thiotetracycline, trefocillin, uracil; CC-1065 and adolaxine, calcetin, pyrazinamide and their synthetic analogs); docalamycin and its synthetic analogs, KW-2189, CBI-TMI or CBI dimer; benzodiazepine dimer or pyrrole benzodiazepine dimer (PBD), tometrazine The following compounds are prohibited from being contaminated with benzodiazepines, indole benzodiazepines, imidazole benzothiazepines, or oxazolidin benzodiazepines; nitrosoureas, including carmustine, lomustine, clostridium chloride, formustine, nimustine, and lamustine; alkyl sulfonates, including betamethasone, styrosine, sulfamethoxazole, and pisufen; triazine or dacarbazine; platinum-containing compounds, including carboplatin, cisplatin, and oxaliplatin; acridine derivatives, such as benzodihydropyranone, carboxone, methotrexate, and uredopa; ethyleneimine and methyl melamine, including hexamethylmelamine, triethylenetriamine, triethylphosphonamide, triethylenethiophosphonamide, and trimethylolpropionic acid; b) Plant alkaloids selected from the following groups of Vinca minor alkaloids: including vincristine, vinblastine, vinorelbine, vinorelbine, and norvincristine; taxanes: paclitaxel, docetaxel, and their analogues; maytansines including DM1, DM2, DM3, DM4, DM5, DM6, DM7, maytansine, salinomycin, and their analogues; cryptophycin (including cryptophycin 1 and cryptophycin 8); epothilone, soft coral alcohol, dimolide, bryophyllactone, sea haretoxin, auristatin, cephalostatin; pancratistatin; sarcodictyin; spongistatin; c) DNA topoisomerase inhibitors: Etoposide-tinib selected from the following groups: including 9-aminocamptothecin, camptothecin, cristatin, doramycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, norsoxamine, retinoid (retinol), teniposide, topotecan, 9-nitrocamptothecin, or RFS 2000; mitomycin and its derivatives; d) Antimetabolites, selected from the following groups [antifolate agents: (DHFR inhibitors (methotrexate, tromethorphan, folate, pteroxate, aminopterin (4-aminobenzoic acid) or other folic acid analogs); IMP dehydrogenase inhibitors: (including mycophenolate mofetil, thiazolinone, ribavirin, EICAR); ribonucleotide reductase inhibitors (including hydroxyurea, deferoxamine)];[Pyrimidine analogues: uracil analogues (including ancitabine, azacitidine, 6-azouracil, capecitabine (Xeloda), carmoflu, cytarabine, dideoxyuridine, deoxyfluorouridine, enoxabin, 5-fluorouracil, fluorouridine, ratitrexed; cytosine analogues (including cytarabine, cytosine arabinoside, fludarabine); purine analogues (including azathioprine, fludarabine, mercaptopurine, thiamine, thioguanine)]; folic acid supplements, florrin; e) Hormone therapy agents: Selected from the following group of receptor antagonists [anti-estrogens (medroxyprogesterone acetate, raloxifene, tamoxifen); LHRH agonists (including gostalin, leuprolide acetate); anti-androgens (including bicalutamide, flutamide, carlusone, betahistelone propionate, epiandrogen, goserelin, leuprolide, metronidazole, nilumid, testrolide, tralostertan and other androgen inhibitors)]; retinoids [vitamin D3 analogs (including CB1093, EB1089, KH1060, cholecalciferol, ergocalciferol); photodynamic therapy agents (including verteporfin, phthalocyanine, photosensitizer Pc4, demethoxy-rubigin A); cytokines (including interferon-α, interferon-γ, tumor necrosis factor (TNF), TNF-containing human proteins)]; f) Kinase inhibitors including BIBW 2992 (anti-EGFR / Erb2), imatinib, gefitinib, guaranatinib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEGFR2), mubritinib, ponatinib, bafetini, bosutinib, cabozantinib, vemodega, iniparib, rusoritinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab, cetuximab, trastuzumab, ranibizumab, panitumumab, ispinib; g) poly(ADP-ribose) polymerase (PARP) inhibitors, such as olaparib, niraparib, iniparib, tarazoparib, veliparib, veliparib, CEP 9722 (Cephalon), E7016 (Eisai), BGB-290 (Beigene), or 3-aminobenzoylamine;h) Antibiotics, including catechins selected from the following groups (cazithromycin, especially cazithromycin γ1, δ1, α1 or β1, dyinmycin, including dyinmycin A and desoxymycin, esperamycin, katimycin, C-1027, madropeptin, or neocazithromycin oseltidine and related chromogen catechin antibiotics), aclacinomysins, actinomycins, atrazomycin, diazoserine, etc. Bleomycin, Carnoycin, Karamycin, Oleomycin, Carcinomacin, Chromomycin, Daclocycline, Daunorubicin, Dedarubicin, 6-Diazon-5-Side-O-L-Demeleucine, Doxorubicin, Moroxydoxorubicin, Cyanomorpholinedoxorubicin, 2-Pyrrolidone Doxorubicin and Deoxydaunorubicin, Epirubicin, Azorubicin, Idarubicin, Macomycin, Nitomycin, Mycophenolic acid, Nogamycin, Olivycin, Peplomycin, Potfiromycin, Purinycin, Quillamycin, Rodolmycin, Streptomycin, Streptozotocin, Tuberculin, Ubenimex, Netostatin, Zolarubicin; i) Polyketides (anthocyanins), bullatacin and bullatacinone; gemcitabine, cyclooxygenase, carfenomib, bortezomib, thalidomide, lenalidomide, pomalidomide, tosedostat, zybrestat, PLX4032, STA-9090, Stimuvax, allovoctin-7, Xegeva, Provenge, Yervoy, isoprene Chemotherapy inhibitors and lovastatin, dopaminergic neurotoxins and 1-methyl-4-phenylpyridinium ion, cell cycle inhibitors (including astrosporins), actinomycins (including actinomycin D, dermatomycin), amatoxins, bleomycin (including bleomycin A2, bleomycin B2, pepromycin), anthracycline antibiotics (including daunorubicin, doxorubicin (adriamycin), idarubicin, epirubicin, pirarubicin, zorubicin, mitoxantrone, MDR inhibitors (such as verapamil), Ca2+ +ATPase inhibitors or carotenoids, histone deacetase inhibitors (including vorinostat, romidesin, pabisostat, valproic acid, mocetinostat (MGCD0103), Belinostat, PCI-24781, entenotide, SB939, resminostat, givinostat, AR-42, CUDC-101, sulforaphane, trachomatis A); celecoxib, glitazones, epigallocatechin gallate, disulfiram, salinosporamide A;Antiadrenergic drugs, such as aminoglutethimide, mitotane, tralostertan, acetylglucuronolactone, aldoxime, aminoacetylpropionic acid, acridine, arabinoside, bestrabucil, bisacodyl, edatraxate, defofamine, mecosine, diazinon, efornithine (DFMO), elfomithine, elemetidine, etoradin, gallium nitrate, cytosine, hydroxyurea, ibandronate, and lentinan. Polysaccharides, chlordamine, mitoxantrone, mitoxantrone, mopudarol, diaminonitroacin, pentostatin, methamidophos, pirarubicin, podophyllic acid, 2-ethylhydrazine, methylbenzylhydrazine; PSK®; guanazine dionepropane; rhizomycin; cizol; spirocyclic germanium; sigmazoic acid; triaminoquinone; trichlorotriethylamine; trichothecene (including T-2 toxin, verrucosporin A, baculosporin A, and anguidine), polyurethane, siRNA, antisense drugs; 2) Autoimmune disease drugs: Cyclosporine, Cyclosporine A, Aminocaproic acid, Azathioprine, Bromocriptine, Chloroquine, Cyclophosphamide, Corticosteroids (including Ancinonide, Betamethasone, Budesonide, Hydrocortisone, Flunisolone, Fluticasone Propionate, Flucolone Danazol, Dexamethasone, Triamcinolone, Beclomethasone Dipropionate), DHEA, Etanercept, Hydroxychloroquine, Infliximab, Meloxicam, Methotrexate, Mycophenolate Mofetil, Prednisone, Sirolimus, Tacrolimus; 3) Anti-infective agents include: ; a) Aminoglycosides: Amikacin, Wuyimycin, Gentamicin (Netilmicin, Sisomicin, Isapamicin), Hygromycin, Kanamycin (Amikacin, Abekaricin, Aminodeoxykanamycin, Dibekacin, Tobramycin), Neomycin (Neomycin B, Paromomycin, Ribomycin), Netilmicin, Spectinomycin, Streptomycin, Tobramycin, Methylstilbestrol; b) Azamides: Chloramphenicol, Chloramphenicol, Flumethoxam, Methoxam; c) Ansarmycins: Gerdmycin, Atrazine; d) Carbapenems: Biapenem, Donipenem, Ertapenem, Imipenem / Cilastatin, Meropenem, Panipenem; e) Cephalosporins: Carbazosporins (carbazosporins), cefaclor, cefaclor, cefadroxil, cefadroxil, cefotaxime, cefalothin or cephalosporins, cefalexin, cefotaxime, cefamandole, cefepime, ceftriaxone, cefotaxime, cefoxitin, cefazolin, cefazolin, cefadroxil, cefadroxil, cefadroxil, cefadroxil, cefadroxil, cefazolin, cefadroxil, cefazolin, cefdavidiana, cefdextromethorphan, cefazolin, cefazolin, cefazolin, cefazolin, cefazolin, cefazolin, cefazolin, cefazolin, cefepime Cefotaxime, cefotaxime, cefodizine, cefnicotinic acid, cefotiam, cefotaxime, cefotaxime, cefotiam, cefazolin, cefalexin, cefimazole, cefpirome, cefpirome, cefpodoxime, cefprozil, cefquinoxime, cefsulfuron-methyl, ceftazidime, cefterenol, cefbuprofen, cefotaxime, ceftriaxone, cefuroxime, cefazolin, cephalosporins (including cefoxitin, cefotetan, cefmetazole), oxcephalosporins (fluoxefaclor, latamoxef). f) Glycopeptides: Bleomycin, Vancomycin (including Orivamphenicol, Tervasin), Teicoplanin (Dabavancin), Ramolane; g) Glycine: Tigecycline; h) β-Lactamase inhibitors: Penicillane (Sulbactam, Tazobactam), Clavulanic acid; i) Lincosamides: Clindamycin, Lincomycin; j) Lipopeptides: Daptomycin, A54145, Calcium-dependent antibiotics (CDA); k) Macrolides: Azithromycin, Quinoerythromycin, Clarithromycin, Dierythromycin, Erythromycin, Fluerythromycin, Josamycin, Ketolides (Telithromycin, Quinoerythromycin), Midecamycin, Meokamycin, Rifampicin, Rifamycin (Rifampin, Rifampicin, Rifabutin, Rifapentine), Rotamycin, Roxithromycin, Spectinomycin, Spiramycin, Tacrolimus (FK506), Acetylosin, Telithromycin; l) Monocyclic β-lactam antibiotics: aztreonam, tegafur; m) Azolidinones: linezolid;n) Penicillins: Amoxicillin, Ampicillin (Pipacillin, Hetacillin, Bamectin, Metancillin, Phthalimicin), Azidocillin, Aloxicillin, Penicillin, Benzathine Penicillin, Phenoxybenzyl Penicillin, Clomicillin, Procaine Penicillin, Carbenicillin (Carinin), O-cloxacillin, Dicloxacillin, Cefazolin, Flucloxacillin, Mecillin (Azamidin Diester), Meropenem, Methicillin, Nafcillin, Oxadiazine, Acetocillin, Penicillin, Necillin, Penicillin, Guaricillin, Phenprocillin, Sulbenicillin, Temoxicillin, Tekacillin; o) Polypeptides: Bacitracin, Polymyxin E, Polymyxin B; p) Quinolones: Araflavin, Barofloxacin, Ciprofloxacin, Clinfloxacin, Daflonaxyl, Difloxacin, Enrofloxacin, Floxin, Garefloxacin, Gatifloxacin, Gemmifloxacin, Gaptafloxacin, Travafloxacin, Levofloxacin, Lomefloxacin, Mebofloxacin, Moxifloxacin, Naflufloxacin, Norfloxacin, Obibixin, Ofloxacin, Pefloxacin, Travafloxacin, Gaptafloxacin, Sitafloxacin, Sparfloxacin, Temafloxacin, Tosufloxacin, Travafloxacin; q) Antibiotics: Punamycin, Quinupordin / Dalfopristin; r) Sulfonamides: Sulfamilone, Prontocillin, Sulfaacetic acid, Sulfamethoxazole, Sulfasalazine, Sulfamethoxazole, Trimethoprim- Sulfamethoxazole (compound sulfamethoxazole); s) Steroid antibiotics: Fusidic acid selected from the following groups; t) Tetracyclines: Doxycycline, Chlortetracycline, Chloramphenicol, Demeclocycline, Lysylcycline, Chlormethoxycycline, Methacycline, Minocycline, Oxytetracycline, Penicillin, Rolicycline, Tetracycline, Glycine (including Tigecycline); u) Other types of antibiotics selected from the following groups: Annonaceae, Arsphenamine, Bacterial Terpene Inhibitors (Bacitracin), DADAL / AR inhibitors (cycloserine), dictyostatin, spongolactone, ilexelose, epothilone, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole, mupirocin, mycolactones, NAM synthesis inhibitors (such as fosfomycin), nitrofurantoin, paclitaxel, picaricin, pyrazinamide, quinupristin / dalfopristin, rifampin, tazobactam-tinidazole, anechoic acid lactone; 4) Antiviral drugs: a) Entry / fusion inhibitors: apraviroc, maraviro, velivirol, gp41 (enfvir), PRO140, CD4 (ibalizumab); b) Integrase inhibitors: raltelavin, ertilavin, globoidnan A; c) Maturation inhibitors: bevirimiloma, vivecon; d) Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir;e) Nucleosides and nucleotides: abacavir, acyclovir, adefovir, amdoxovir, apricitabine, brivudine, cidofovir clavudine, dexavistatin, doxavistatin (DDI), apricitabine, emtricitabine (FTC), entecavir, famciclovir, fluorouracil (5-FU), 3'-fluorosubstituted 2',3'-dideoxynucleoside derivatives (e.g., 3'-fluoro-2',3'-dideoxythymidine (FLT) and 3'-fluoro-2 ',3'-Dideoxy (FLG) fomivir, ganciclovir, idoxuridine, lamivudine (3TC), L-nucleoside (including β-L-thymidine, β-L-2'-deoxycytidine), penciclovir, Racivir, ribavirin, stampidine, stavudine (d4T), tarivilin (veramidine), telbivudine, tenofovir, valacyclovir trifluorothymidine, valganciclovir, zalcitabine (DDC), zidovudine (AZT); f) Non-nucleoside analogues: Amantadine, Atenviridine, Capvirine, Diarylpyrimidines (Etravirine, Rilpivirine), Deraviridine, Docosanol, Emivirine, Efavirenz, Phosphoric Acid, Imiquimod, Interferon Alpha, Loviramide, Lodeadenosine, Thiapazole, Nevirapine, NOV-205, Pegylated Interferon Alpha, Podophyllotoxin, Rifampin, Amantadine, Requimod (R-848), Amantadine; g) Protease inhibitors: Ampravir, Atazanavir, Boseravir, Darunavir, Fosanavir, Indinavir, Lopinavir, Nefernavir, Pleconaril, Ritonavir, Saquinavir, Telapirvir (VX-950), Telanavir; h) Other types of antiviral drugs: Anti-enzymes, Arbidol, Calanolides A. cyanobacterium, cyanobacterium-N, diarylpyrimidine, epigallocatechin gallate (EGCG), sodium phosphonoformate, griseofulvin, tarivilin (veramidine), hydroxyurea, KP-1461, mitefonine, precocinone, synthesis inhibitors, ribavirin, seliciclib; (5) or pharmaceutically acceptable salts, hydrates or hydrated salts thereof; or polycrystalline structures of such compounds; or isomers, optical isomers, racemates, non-mirror image isomers or mirror image isomers thereof.

16. The pharmaceutical composition of claim 14, wherein the synergist is selected from the group consisting of one or more of the following drugs: abatacept, abiraterone acetate, acetaminophen / hydrocodone, adalimumab, ADXS31-142, ADXS-HER2, afatinib maleate, alenmab, trastuzumab, amphetamine / dextromethorphan, anastrozole, atazanavir, atezolizumab, atorvastatin, axitinib, beline, BCG Live, bevacizumab, cabazitaxel, cabozantin, bexarotin, blinatumomab, bortezomib, bosutinib, brentuximab Vedotin, Budesonide, Budesonide / Formoterol, Buprenorphine, Capecitabine, Carfilzomib, Celecoxib, Cetuximab, Cyclosporine, Cinacalcet, Crizotinib, Dabigatran, Dabrafenib, Afadabetin, Derrenavir, Imatinib Mesylate, Dasatinib, Denisil Interleukin, Denosema, Sodium Valproate, Dexlansoprazole, Dextromethorphan, Dinutuximab, Doxycycline, Duloxetine, Emtricitabine / Rilpivirine / Tenofovir Disoproxil Fumarate, Emtricitabine / Tenofovir / Efavirenz, Enoxaparin, Enzalutamide, Epoxetine Afa, Erlotinib Esomeprazole, Ezetimibe, Ezetimibe / Simvastatin, Fenofibrate, Filgrastim, Fingolimod, Fluticasone Propionate, Fluticasone / Sameterol, Fulvestrant, Gefitinib, Glatiramer, Goserelin Acetate, Imatinib, Tiimomab, Ibrutinib, Insulin Aspart, Insulin Detemir, Insulin Glargine, Insulin Lispro, Interferon β1a, Interferon β1b, Lapatinib, Ipilimumab, Ipratropium Bromide / Salbutamol, Lanleptide Acetate, Lenalidomide, Combined Benzoylsulfonate, Letrozole, Levothyroxine, Levothyroxine, Lidocaine, Linezolid, LiraglutideMEDI4736, Memantine, Methyl acetate, Metoprolol, Modafinil, Mometasone, Nirotinib, Nivolumab, Olfamumab, Olitazolizumab, Pazopanib, Pembrolizumab, Pemetrexed, Pertuzumab, Pneumococcal conjugate vaccine, Pomalidomide, Pregabalin, Quetiapine, Rabeprazole radium chloride 223, Raloxifene, Ralcifen, Ramirez, Ranibizumab, Regorafenib, Rituximab, Rivaroxaban, Romidixin, Rosuvastatin, Russoteinib phosphate, Salbutamol, Severamine, Sildenafil, Silt uximab, sitagliptin, sitagliptin / metformin, sofenacin, sorafenib, sunitinib, tadalafil, tamoxifen, telaprevir, tesimolimus, tenofovir / emtricitabine, testosterone gel, thalidomide (Talidex), tiotropium bromide, toremifene, trametinib, trastuzumab, tretinoin, eutecticumab, valsartan, vandetanib, vemurafenib, vorinostat, aflibercept, Zostavax and their analogues, derivatives, pharmaceutically acceptable salts, carriers, diluents or excipients, or combinations thereof.

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