Camptothecin derivatives, methods for preparing the same, and their use

Abstract

The present invention provides a novel camptothecin-derived compound, a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, and methods for their preparation and use. The compound has a structure represented by formula (I) and has good anti-cancer activity. [Chemical Formula 1] TIFF2025524170000094.tif44170

Description

[Technical Field]

[0001] This invention relates to the pharmaceutical field, and more specifically to a series of novel camptothecin derivatives having a new structure, as well as methods for preparing and using them. [Background technology]

[0002] Camptothecin (CPT) is a natural product first extracted from the Chinese plant *Hydrophila zebrina* by American scientists Wall and Wani. It is a water-insoluble, cytotoxic quinoline alkaloid with broad antitumor activity. Its mechanism of action is thought to involve binding to topoisomerase Top1 and DNA to form a complex, inhibiting Top1 activity, blocking DNA replication and protein synthesis, and leading to apoptosis. Camptothecin has shown significant therapeutic effects against gastric cancer, esophageal cancer, cardia cancer, colon cancer, rectal cancer, primary liver cancer, acute and chronic granulocytic leukemia, choriocarcinoma, lung cancer, and bladder cancer. Despite their excellent antitumor activity, only two camptothecin compounds have been developed globally as antitumor agents: irinotecan, useful for colorectal cancer and small cell lung cancer (SCLC), and topotecan, useful for lung cancer and ovarian cancer. Another camptothecin derivative, beletecan, is approved only in South Korea as a treatment for SCLC and ovarian cancer. Camptothecin's cytotoxicity, poor solubility, and drug resistance have hindered its broader application in tumor treatment.

[0003] Therefore, there is a need to develop low-toxicity, water-soluble, and drug-resistant camptothecin derivative drugs, which have broad application potential. [Overview of the project] [Problems that the invention aims to solve]

[0004] One object of the present invention is to provide a camptothecin derivative having a novel structure that exhibits anticancer activity.

Means for Solving the Problem

[0005] One aspect of the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof.

Chemical formula

[0006] Wherein, R is -CH2NR1R2, wherein R1 and R2 are each independently hydrogen, a hydroxy group, a C1-C6 alkyl group, a C1-C6 alkoxy group, a C3-C6 cycloalkyl group, a 4-8 member heterocycloalkyl group, a 4-8 member aryl group or -NR a R b selected from, and the C1-C6 alkyl group, C1-C6 alkoxy group, C3-C6 cycloalkyl group, 4-8 member heterocycloalkyl group, 4-8 member aryl group or -NR a R b is optionally substituted with a substituent selected from halogen, a hydroxy group, an amino group, a carboxyl group, a C1-C6 alkoxy group, a C1-C6 alkoxycarbonyl group, -O-(C1-C3 alkyl)-NR a R b group, or a 4-8 member heterocycloalkyl group, provided that R1 and R2 are not simultaneously hydrogen or methyl, or R1 and R2 together with the N atom to which they are attached form a 4-8 member cycloalkyl group, a 4-8 member heterocycloalkyl group, a 4-8 member aryl group or a 9-12 member azaspiroalkyl group, and the 4-8 member cycloalkyl group, 4-8 member heterocycloalkyl group, 4-8 member aryl group or 9-12 member azaspiroalkyl group is optionally a hydroxy group, an amino group, a carboxyl group, a C1-C6 alkoxy group, a C1-C6 alkoxycarbonyl group, a C1-C3 alkyl group, a C1-C3 alkoxy group, -O-NH2, -NR a R b or -C(O)R c selected from substituents, or R and the hydroxyl group in its ortho position, together with the C atom on the benzene ring linked to them, form a 4-8 membered heterocycloalkyl group, and the 4-8 membered heterocycloalkyl group can optionally be a C1-C6 alkyl group, a C1-C6 alkoxy group, a C3-C6 cycloalkyl group, a 4-8 membered heterocycloalkyl group, or -NR a R b Alternatively, it may be substituted with substituents selected from 4-8 member heterocycloalkyl groups, and the C1-C6 alkyl groups, C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, 4-8 member heterocycloalkyl groups, -NR a R b Alternatively, a 4- to 8-membered heterocycloalkyl group may be substituted with substituents selected from halogens, hydroxyl groups, amino groups, carboxyl groups, C1-C6 alkoxy groups, or C1-C6 alkoxycarbonyl groups, provided that at least one of R1 and R2 is H.

[0007] R a and R b Each of these is independently selected from H, a C1-C6 alkyl group, or a C1-C6 alkoxycarbonyl group.

[0008] R c This is selected from an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C3-C6 cycloalkyl group, or a hydroxyl group-substituted C1-C3 alkyl group.

[0009] In one embodiment, in the compound of formula (I), R is -CH2NR1R2, where R1 and R2 are independently hydrogen, a hydroxyl group, a C1-C6 alkyl group, a C1-C6 alkoxy group, a C3-C6 cycloalkyl group, a 4-8 member heterocycloalkyl group, or -NR a R b Selected from the above, the C1-C6 alkyl group may optionally be a halogen, hydroxyl group, amino group, carboxyl group, C1-C6 alkoxycarbonyl group, or -O-(C1-C3 alkyl)-NR a R b, or substituted with a substituent selected from 4-8 membered heterocycloalkyl groups, where the C3-C6 cycloalkyl group or 4-8 membered heterocycloalkyl group is optionally substituted with a substituent selected from a hydroxyl group, a C1-C3 alkoxy group, or a C1-C6 alkoxycarbonyl group, provided that R1 and R2 are not simultaneously hydrogen or methyl groups, or R1 and R2, together with the N atoms linked to them, form a 4-8 member heterocycloalkyl group or a 9-12 member azaspiroalkyl group, and the 4-8 member heterocycloalkyl group may optionally contain a hydroxyl group, an amino group, a carboxyl group, a C1-C6 alkoxycarbonyl group, a C1-C3 alkyl group, a C1-C3 alkoxy group, an -O-NH2 group, or a -C(O)R c Substituted with a substituent selected from, or R and the hydroxyl group in its ortho position, together with the C atom on the benzene ring linked to them, form a 4- to 8-membered heterocycloalkyl group, and the 4- to 8-membered heterocycloalkyl group is optionally substituted with substituents selected from halogen-substituted C1-C6 alkyl groups, C3-C6 cycloalkyl groups, or 4- to 8-membered heterocycloalkyl groups, provided that at least one of R1 and R2 is H.

[0010] In one embodiment, R a and R b Each of these is independently selected from H, a methyl group, or a tert-butoxycarbonyl group.

[0011] In one embodiment, R c This is selected from an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C3-C6 cycloalkyl group, or a hydroxy-substituted C1-C3 alkyl group.

[0012] In one embodiment, in the compound of formula (I), R is -CH2NHR1, and R1 is hydrogen, a hydroxyl group, a C1-C6 alkyl group, a C1-C6 alkoxy group, a C3-C6 cycloalkyl group, a 4-8 member heterocycloalkyl group, or -NR a R bThe C1-C6 alkyl group is selected from the following, and the C1-C6 alkyl group may optionally be a halogen, a hydroxyl group, an amino group, a carboxyl group, a C1-C6 alkoxycarbonyl group, or an -O-(C1-C3 alkyl)-NR a R b , or substituted with a substituent selected from 4-8 membered heterocycloalkyl groups, wherein the C3-C6 cycloalkyl group or 4-8 membered heterocycloalkyl group is optionally substituted with a substituent selected from a hydroxyl group, a C1-C3 alkoxy group, or a C1-C6 alkoxycarbonyl group, provided that R1 is not hydrogen, or R and the hydroxyl group in its ortho position, together with the C atom on the benzene ring linked to them, form a 4- to 8-membered heterocycloalkyl group, and the 4- to 8-membered heterocycloalkyl group may be optionally substituted with substituents selected from halogen-substituted C1-C6 alkyl groups, C3-C6 cycloalkyl groups, or 4- to 8-membered heterocycloalkyl groups.

[0013] In one embodiment, R a and R b Each of these is independently selected from H, a methyl group, or a tert-butoxycarbonyl group.

[0014] In one embodiment, R1 and R2 are independently hydrogen, a hydroxyl group, a methyl group, an ethyl group, a propyl group, an isopropyl group, a methoxy group, an ethoxy group, a propoxy group, a cyclopropyl group, a cyclopentyl group, a cyclohexyl group, a 5-membered or 6-membered heterocycloalkyl group containing 1 to 2 N atoms as ring atoms, a 5-membered or 6-membered heterocycloalkyl group containing 1 to 3 heteroatoms selected from N or O as ring atoms, or -NR a R b Selected from, where the methyl group, ethyl group, propyl group, or isopropyl group may optionally be a halogen, hydroxyl group, amino group, carboxyl group, methoxycarbonyl group, ethoxycarbonyl group, tert-butylalkoxycarbonyl group, or methoxy-NR a R b Ethoxy-NR a R bThe 5-membered or 6-membered heterocycloalkyl group, which is substituted with a substituent selected from a pyrrolidinyl group, a piperidinyl group, or a piperazinyl group, and contains a cyclopropyl group, a cyclopentyl group, a cyclohexyl group, or 1 to 2 N atoms as ring atoms, or the 5-membered or 6-membered heterocycloalkyl group, which contains 1 to 3 heteroatoms selected from N or O as ring atoms, may optionally be substituted with a substituent selected from a hydroxyl group, a methoxy group, an ethoxy group, a propoxy group, or a methoxycarbonyl group, an ethoxycarbonyl group, or a tert-butylalkoxycarbonyl group, provided that R1 and R2 are not simultaneously hydrogen or methyl groups.

[0015] In one embodiment, R1 and R2, together with the N atom linked to them, form a pyrrolidinyl group, piperidinyl group, piperazinyl group, or 11-membered azaspiroalkyl group, and the pyrrolidinyl group, piperidinyl group, piperazinyl group, or 11-membered azaspiroalkyl group may optionally be a hydroxyl group, amino group, carboxyl group, methoxycarbonyl group, ethoxycarbonyl group, tert-butylalkoxycarbonyl group, methyl group, ethyl group, propyl group, isopropyl group, methoxy group, ethoxy group, propoxy group, -O-NH2, or -C(O)R c It is replaced with a substituent selected from the following.

[0016] In one embodiment, R and the hydroxyl group in its ortho position, together with the C atom on the adjacent benzene ring, form a pyrrolidinyl group, piperidinyl group, piperazinyl group, or oxazinyl group, and the pyrrolidinyl group, piperidinyl group, piperazinyl group, or oxazinyl group is optionally substituted with substituents selected from a methyl group, ethyl group, propyl group, isopropyl group, cyclopropyl group, cyclopentyl group, cyclohexyl group, pyrrolidinyl group, piperidinyl group, piperazinyl group, or oxazinyl group.

[0017] In one embodiment, in the compound of formula (I), R1 and R2 are independently hydrogen, hydroxyl group, amino group, methyl group, ethyl group, isopropyl group, methoxy group, ethoxy group, trifluoroethyl group, cyclopropyl group, -(CH2)2OH, and -(CH2) 2-4 Selected from NH2, -CH(CH3)COOH, -CH(CH3)CH2OCH3, -CH(CH3)CH2OH, -(CH2)2CH3, -NHBoc, N(CH3)Boc, -CH(CH3)Boc, -(CH2)2O(CH2)2NHBoc, and the group shown in the following formula, [ka] However, this is conditional on R1 and R2 not being simultaneously hydrogen or methyl groups, or R1 and R2, together with the N atoms linked to them, form pyrrolidine, piperidine, piperazine, or 3,9-diazaspirondecane (for example, the compounds listed below). [ka] The aforementioned pyrrolidine, piperidine, piperazine, or 3,9-diazaspiloundecane may optionally contain a hydroxyl group, amino group, methyl group, methoxy group, carboxyl group, tert-butoxycarbonyl group, -O-NH2, or -C(O)R c Substituted with a substituent selected from, or R and the hydroxyl group in its ortho position, together with the C atom on the adjacent benzene ring, form 1,3-oxadinane, and the 1,3-oxadinane is optionally substituted with a substituent selected from a trifluoromethyl group, a cyclopropyl group, or morpholine-4-yl, provided that at least one of R1 and R2 is H. c The group is selected from an amino group, a methyl group, a methoxy group, a hydroxyethyl group, or a cyclopropyl group.

[0018] In one embodiment, R and the hydroxyl group in its ortho position, together with the C atom on the adjacent benzene ring, form a group represented by the following formula: [ka] The group represented by the above formula may be optionally substituted with a substituent selected from C1-C6 alkyl groups or 4-8 membered heterocycloalkyl groups.

[0019] In one embodiment, R and the hydroxyl group in its ortho position, together with the C atom on the adjacent benzene ring, form a group represented by the following formula: [ka] The group represented by the above formula may be optionally substituted with a substituent selected from a trifluoromethyl group, a cyclopropyl group, or morpholine-4-yl, provided that at least one of R1 and R2 is H.

[0020] In one embodiment, R1 and R2, together with the N atoms linked to them, form pyrrolidine, piperidine, piperazine, or 3,9-diazaspiloundecane, and the pyrrolidine, piperidine, piperazine, or 3,9-diazaspiloundecane is optionally substituted with substituents selected from an amino group, a methoxy group, -O-NH2, a carboxyl group, a tert-butoxycarbonyl group, -C(O)-NH2, -C(O)OCH3, -C(O)CH3, -C(O)-cyclopropyl group, or -C(O)CH2OH.

[0021] In one embodiment, R is -CH2NHR1, and R1 is hydrogen, hydroxyl group, amino group, methyl group, ethyl group, isopropyl group, methoxy group, ethoxy group, trifluoroethyl group, cyclopropyl group, -(CH2)2OH, -(CH2) 2-4 Selected from NH2, -CH(CH3)COOH, -CH(CH3)CH2OCH3, -CH(CH3)CH2OH, -(CH2)2CH3, -NHBoc, N(CH3)Boc, -CH(CH3)Boc, -(CH2)2O(CH2)2NHBoc, and the group shown in the following formula, [ka] However, this is conditional on R1 not being hydrogen, or R and the hydroxyl group in its ortho position, together with the C atom on the adjacent benzene ring, form a group represented by the following formula: [ka] The group represented by the above formula may be optionally substituted with a substituent selected from a trifluoromethyl group, a cyclopropyl group, or morpholine-4-yl.

[0022] In one embodiment, R and the hydroxyl group in its ortho position, together with the C atom on the adjacent benzene ring, form a group represented by the following formula: [ka] The group represented by the above formula may optionally be substituted with a trifluoromethyl group or a morpholine-4-yl substituent.

[0023] In one embodiment, R is selected from the following structures. [ka]

[0024] In one embodiment, R is selected from the following structures. [ka]

[0025] In one embodiment, R is selected from the following structures. [ka]

[0026] In the present invention, in the above compound, R does not include structures such as -CH2NHCH2CH2OH, -CH2NHCH2CH2N(CH3)2, and the group represented by the following formula. [ka] Furthermore, the compound of formula (I) does not contain the following compounds. [ka]

[0027] In one embodiment, the compound of formula (I) is a compound having the structure of formula (II). [ka]

[0028] During the ceremony, The Y ring is selected from a 4-8 member nitrogen-containing saturated heteroring, a 4-8 member nitrogen-containing aromatic ring, or a 9-12 member nitrogen-containing saturated spiroring, and the 4-8 member nitrogen-containing saturated heteroring, 4-8 member nitrogen-containing aromatic ring, or 9-12 member nitrogen-containing saturated spiroring may optionally be a hydroxyl group, an amino group, a C1-C6 alkoxycarbonyl group, a C1-C3 alkyl group, a C1-C3 alkoxy group, -O-NH2, or -C(O)R c The C1-C6 alkoxycarbonyl group is further substituted with a substituent selected from the above, and preferably the C1-C6 alkoxycarbonyl group is a tert-butoxycarbonyl group.

[0029] Preferably, the Y ring is selected from a 4-8 member nitrogen-containing saturated ring or a 9-12 member nitrogen-containing saturated spiro ring, and the 4-8 member nitrogen-containing saturated ring may optionally be a hydroxyl group, an amino group, a C1-C6 alkoxycarbonyl group, a C1-C3 alkyl group, a C1-C3 alkoxy group, -O-NH2, or -C(O)R c The C1-C6 alkoxycarbonyl group is substituted with a substituent selected from the above, and preferably the C1-C6 alkoxycarbonyl group is a tert-butoxycarbonyl group.

[0030] R c This is selected from a hydroxyl group, an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C3-C6 cycloalkyl group, or a hydroxy-substituted C1-C3 alkyl group.

[0031] Preferably, Rc The group is selected from a hydroxyl group, an amino group, a methyl group, a methoxy group, a hydroxymethyl group, a tert-butyloxy group, or a cyclopropyl group.

[0032] In one embodiment, the compound of formula (II) is a compound having the structure of formula (IIa), formula (IIb), or formula (IIc). [ka]

[0033] Here, R c This is selected from a hydroxyl group, an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C3-C6 cycloalkyl group, or a hydroxy-substituted C1-C3 alkyl group.

[0034] Preferably, R c The group is selected from a hydroxyl group, an amino group, a methyl group, a methoxy group, a hydroxymethyl group, a tert-butyloxy group, or a cyclopropyl group. [ka]

[0035] During the ceremony, The Y ring is selected from a 4-8 member nitrogen-containing saturated heteroring, a 4-8 member nitrogen-containing aromatic ring, or a 9-12 member nitrogen-containing saturated spiroring, and the 4-8 member nitrogen-containing saturated heteroring, 4-8 member nitrogen-containing aromatic ring, or 9-12 member nitrogen-containing saturated spiroring may optionally consist of a hydroxyl group, an amino group, a C1-C6 alkoxycarbonyl group, a C1-C3 alkyl group, a C1-C3 alkoxy group, -O-NH2, or -C(O)R c The C1-C6 alkoxycarbonyl group is further substituted with a substituent selected from the above, and preferably the C1-C6 alkoxycarbonyl group is a tert-butoxycarbonyl group.

[0036] Here, R c It is defined similarly to the compound of formula (I).

[0037] Preferably, the Y ring is selected from a 4-8 member nitrogen-containing saturated ring or a 9-12 member nitrogen-containing saturated spiro ring, and the 4-8 member nitrogen-containing saturated ring may optionally be a hydroxyl group, an amino group, a C1-C6 alkoxycarbonyl group, a C1-C3 alkyl group, a C1-C3 alkoxy group, -O-NH2, or -C(O)R c The C1-C6 alkoxycarbonyl group is substituted with a substituent selected from the above, and preferably the C1-C6 alkoxycarbonyl group is a tert-butoxycarbonyl group.

[0038] R c ' is selected from a hydroxyl group, an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C3-C6 cycloalkyl group, or a hydroxy-substituted C1-C3 alkyl group.

[0039] Preferably, R c ' is selected from a hydroxyl group, amino group, methyl group, methoxy group, hydroxymethyl group, tert-butyloxy group, or cyclopropyl group.

[0040] In one embodiment, the compound of formula (II) is a compound having the structure of formula (IIa), formula (IIb), or formula (IIc), [ka] Here, R c ' is selected from a hydroxyl group, an amino group, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C3-C6 cycloalkyl group, or a hydroxy-substituted C1-C3 alkyl group.

[0041] Preferably, R c ' is selected from a hydroxyl group, amino group, methyl group, methoxy group, hydroxymethyl group, tert-butyloxy group, or cyclopropyl group.

[0042] This invention provides the following compounds. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0043] Another aspect of the present invention provides a method for preparing the above compound, comprising steps selected from the following reaction pathways.

[0044] Reaction pathway 1: [ka] 10-hydroxycamptothecin, formaldehyde or paraformaldehyde, and an amine compound are subjected to a Mannich reaction at the 9-position to obtain the compound of formula (I).

[0045] Here, the amine compound is NHR1R2, where R, R1, and R2 are defined as described above.

[0046] Reaction pathway 2: [ka] 9-Formyl-10-hydroxycamptothecin, an amine compound, and sodium cyanoborohydride are subjected to a reductive amination reaction to obtain the compound of formula (I).

[0047] Here, the amine compound is NHR1R2, where R, R1, and R2 are defined as described above.

[0048] In one embodiment, 9-formyl-10-hydroxycamptothecin is obtained by reacting 10-hydroxycamptothecin with hexamethylenetetramine.

[0049] In one embodiment, in the Mannich reaction described in reaction route 1, the amount of paraformaldehyde or formaldehyde used is 1.0 to 5.0 molar equivalents, preferably 1.0 to 3.0 molar equivalents, and more preferably 1.2 to 2.2 molar equivalents.

[0050] Preferably, in reaction pathway 1, the amount of amine compound used is 1.0 to 5.0 molar equivalents, more preferably 1.0 to 2.0 molar equivalents, and more preferably 1.2 to 1.6 molar equivalents.

[0051] In one embodiment, in the reductive amination reaction described in reaction pathway 2, the reducing agent is sodium borocyanohydride.

[0052] Preferably, in reaction pathway 2, the amount of amine compound used is 1.0 to 3.0 molar equivalents, and more preferably 1.0 to 1.2 molar equivalents.

[0053] Preferably, the amount of sodium cyanoborohydride used is 1.0 to 3.0 molar equivalents, more preferably 1.0 to 2.0 molar equivalents, and even more preferably 1.2 to 1.5 molar equivalents.

[0054] Another aspect of the present invention provides an antibody-drug conjugate compound comprising a small molecule drug and a linker antibody, wherein the small molecule drug is the above compound or a pharmaceutically acceptable salt, stereoisomer, or prodrug thereof.

[0055] Another aspect of the present invention provides a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt thereof, a stereoisomer, a prodrug, or the above antibody-drug conjugate.

[0056] The aforementioned pharmaceutical composition further includes pharmaceutically acceptable adjuvants.

[0057] Another aspect of the present invention provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof, stereoisomer or prodrug, antibody-drug conjugate or pharmaceutical composition in the preparation of a drug for treating cancer.

[0058] In a particular manner, the cancers include gastric cancer, esophageal cancer, cardia cancer, breast cancer, ovarian cancer, colon cancer, rectal cancer, primary liver cancer, acute and chronic granulocytic leukemia, choriocarcinoma, lung cancer, bladder cancer, intestinal cancer, and small cell lung cancer, and preferably, the cancers are esophageal cancer, breast cancer, and gastric cancer.

[0059] Another aspect of the present invention provides a method for treating cancer, comprising administering the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer, prodrug, antibody-drug conjugate, or pharmaceutical composition thereof to a patient in need.

[0060] In one embodiment, the dosage of the above compound or its pharmaceutically acceptable salt, stereoisomer, prodrug, antibody-drug conjugate, or pharmaceutical composition is a therapeutically effective amount.

[0061] The camptothecin derivative compounds provided by the present invention exhibit significant inhibitory activity against cancer cells such as esophageal cancer cells OE33, mammary gland cancer cells SK-BR-3, and gastric cancer cells NCI-N87.

[0062] Notably, the present invention also provides a method for preparing the aforementioned compounds in remarkably high yields. [Modes for carrying out the invention]

[0063] I. Definition In this invention, the scientific and technical terms used herein have meanings that are generally understood by those skilled in the art, unless otherwise specified. Furthermore, the related terms and experimental procedures used herein are terms and steps that are widely used in their respective fields. In addition, to better understand this invention, definitions and explanations of the related terms are provided below.

[0064] As used herein, and unless otherwise specified, the terms “include,” “contain,” “have,” and “contain,” including their grammatically equivalent forms, should generally be understood as open and non-restrictive, and not, for example, preclude other elements or steps not listed.

[0065] The compounds according to the present invention may be asymmetric and, for example, may have one or more stereoisomers. Unless otherwise specified, this includes all stereoisomers such as enantiomers and diastereomers. The stereoisomers include geometric isomers (e.g., cis and trans structures) and optical isomers (e.g., enantiomers), and therapeutic agents consisting of elements, racemates, racemic mixtures, and pharmaceutically acceptable salts thereof. Compounds containing asymmetric carbon atoms according to the present invention can be isolated in optically active pure form or racemic form. The optically active pure form can be isolated from racemic mixtures or synthesized using chiral starting materials or chiral reagents. Racemates, diastereomers, and enantiomers are included within the scope of the present invention.

[0066] The compounds of the present invention also include tautomers. The tautomer form arises from the exchange of a single bond with an adjacent double bond, both involving the transfer of one proton.

[0067] Numerical ranges in this specification refer to individual integers within a specified range. For example, "C1-C6" means that the group may have one, two, three, four, five, or six carbon atoms, and "C3-C6" means that the group may have three, four, five, or six carbon atoms.

[0068] The term "substituted" means that any one or more hydrogen atoms on a particular atom or group are replaced by a substituent, provided that the valence of that atom or group remains normal and the resulting compound is stable. Unless otherwise specified, the type and number of substituents are arbitrary, provided they are chemically feasible.

[0069] If any variable (e.g., Rn) appears one or more times in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted with 1 to 5 Rs, that group may be arbitrarily substituted with up to 5 Rs, and in each case, R has an independent choice. Furthermore, combinations of substituents and / or their variants are only permissible if such combinations result in a stable compound.

[0070] The term "alkyl group" refers to a saturated aliphatic hydrocarbon group, which is a linear or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-restrictive examples include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, tert-butyl group, sec-butyl group, n-pentyl group, 1,1-dimethylpropyl group, 1,2-dimethylpropyl group, 2,2-dimethylpropyl group, 1-ethylpropyl group, 2-methylbutyl group, 3-methylbutyl group, n-hexyl group, 1-ethyl-2-methylpropyl group, 1,1,2-trimethylpropyl group, 1,1-dimethylbutyl group, 1,2-dimethylbutyl group, 2,2-dimethylbutyl group, 1,3-dimethylbutyl group, 2-ethylbutyl group, 2-methylpentyl group, 3-methylpentyl group, 4-methylpentyl group, 2,3-dimethylbutyl group, n-heptyl group, 2-methylhexyl group, 3-methylhexyl group, 4-methylhexyl group, 5-methylhexyl This includes groups such as 2,3-dimethylpentyl group, 2,4-dimethylpentyl group, 2,2-dimethylpentyl group, 3,3-dimethylpentyl group, 2-ethylpentyl group, 3-ethylpentyl group, n-octyl group, 2,3-dimethylhexyl group, 2,4-dimethylhexyl group, 2,5-dimethylhexyl group, 2,2-dimethylhexyl group, 3,3-dimethylhexyl group, 4,4-dimethylhexyl group, 2-ethylhexyl group, 3-ethylhexyl group, 4-ethylhexyl group, 2-methyl-2-ethylpentyl group, 2-methyl-3-ethylpentyl group, n-nonyl group, 2-methyl-2-ethylhexyl group, 2-methyl-3-ethylhexyl group, 2,2-diethylpentyl group, n-decyl group, 2,2-diethylhexyl group, 2,2-diethylhexyl group, and various branched isomers thereof.More preferably, the lower alkyl group contains 1 to 6 carbon atoms, and non-restrictive examples include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, tert-butyl group, sec-butyl group, n-pentyl group, 1,1-dimethylpropyl group, 1,2-dimethylpropyl group, 2,2-dimethylpropyl group, 1-ethylpropyl group, 2-methylbutyl group, 3-methylbutyl group, n-hexyl group, 1-ethyl-2-methylpropyl group, 1,1,2-trimethylpropyl group, 1,1-dimethylbutyl group, 1,2-dimethylbutyl group, 2,2-dimethylbutyl group, 1,3-dimethylbutyl group, 2-ethylbutyl group, 2-methylpentyl group, 3-methylpentyl group, 4-methylpentyl group, 2,3-dimethylbutyl group, and the like. The alkyl group may be substituted or unsubstituted, and if substituted, the substituent may be substituted at any available bond site, and the substituent is preferably one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, mercapto groups, hydroxyl groups, nitro groups, cyano groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, heterocycloalkylthio groups, oxo groups, carboxyl groups, or carboxylic acid ester groups. In the present invention, preferably, the alkyl group is methyl, ethyl, isopropyl, tert-butyl, haloalkyl group, deuterium alkyl group, alkoxy-substituted alkyl group, and hydroxyl-substituted alkyl group.

[0071] The term "heterocyclyl group" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, of which one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O)m (where m is an integer from 0 to 2), but which do not contain -OO-, -OS-, or -SS- ring portions, and the remaining ring atoms are carbon. Preferably, it is a 3 to 8-membered heterocyclyl containing 3 to 12 ring atoms, of which 1 to 4 are heteroatoms, more preferably 3 to 8 ring atoms, most preferably 3 to 8 ring atoms, and even more preferably 1 to 3 nitrogen atoms, which are optionally substituted with 1 to 2 oxygen atoms, sulfur atoms, or oxo groups, and includes nitrogen-containing monocyclic heterocyclyl groups, nitrogen-containing spiroheterocyclyl groups, or nitrogen-containing condensed heterocyclyl groups.

[0072] The term "heterocycloalkyl group" refers to a saturated ring substituent containing heteroatoms on a ring skeleton, where one or more (preferably 1 to 4, 1 to 3, or 1 to 2) ring atoms are heteroatoms selected from N, O, and S.

[0073] The term "aryl group" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably a 6- to 12-membered group such as a phenyl group or a naphthyl group.

[0074] The aryl group may be substituted or unsubstituted. If substituted, the substituent is preferably one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, sulfhydryl groups, hydrogen groups, nitro groups, cyano groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, heterocycloalkylthio groups, carboxyl groups, or carboxylic acid ester groups.

[0075] The term "alkoxy group" refers to -O-(alkyl group) and -O-(unsubstituted cycloalkyl group), and the definition of alkyl group is as described above. Non-restrictive examples of alkoxy groups include methoxy group, ethoxy group, propoxy group, butoxy group, cyclopropoxy group, cyclobutoxy group, cyclopentyloxy group, and cyclohexyloxy group. Alkoxy groups may be optionally substituted or unsubstituted, and if substituted, the substituent is preferably one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylthio groups, alkylamino groups, halogens, sulfhydryl groups, hydrogen groups, nitro groups, chloro groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkoxy groups, heterocycloalkoxy groups, cycloalkylthio groups, heterocycloalkylthio groups, carboxylic acid groups, or carboxylic acid ester groups.

[0076] All hydrogen atoms described in this invention can be substituted with their isotope, deuterium.

[0077] "Optional" or "optionally" means that the event or environment described thereafter may occur, but does not necessarily occur, and the description includes cases where the event or situation occurs and cases where it does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may be present, but is not required, and the description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0078] "Substituting" means that one or more hydrogen atoms in the group, preferably up to five, more preferably one to three, are independently substituted with a corresponding number of substituents. Substituents can only exist in chemically possible positions, and it is obvious to those skilled in the art that possible or impossible substitutions can be determined without undue effort (by experiment or theory). For example, an amino group or hydroxyl group with free hydrogen may become unstable when bonded to a carbon atom with an unsaturated (e.g., alkene) bond.

[0079] The following symbols represent chemical bonding sites. [ka]

[0080] Drug or pharmaceutical composition In this specification, "pharmaceutically acceptable salt" means a salt formed with a corresponding amine compound and an inorganic or organic acid, or a salt formed with a corresponding carboxylic acid compound and an alkali metal or alkaline earth metal, or a salt formed with a corresponding carboxylic acid compound and an organic amine. Here, inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid. Organic acids include, but are not limited to, acetic acid, propionic acid, butyric acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, oxalic acid, succinic acid, lactic acid, citric acid, succinic acid, gluconic acid, maleic acid, fumaric acid, and tartaric acid. Alkali metal or alkaline earth metal salts include, but are not limited to, sodium salts, potassium salts, calcium salts, magnesium salts, etc. Organic amine salts include, but are not limited to, salts made from ammonia, methylamine, ethylamine, propylamine, isopropylamine, dimethylamine, diethylamine, trimethylamine, triethylamine, tert-butylamine, ethylenediamine, ethanolamine, diethanolamine, triethanolamine, morpholine, piperidine, piperazine, amino acids, etc.

[0081] In this specification, "precursor" refers to a compound that, after being taken into the human body by an appropriate administration method, undergoes metabolism or simple chemical changes in the patient's body to be converted into the form of the compound contained in General Formula 1 of the present invention and its corresponding salt. Precursors of the compound include, but are not limited to, various forms such as carboxylic acid esters, carbonate esters, phosphate esters, sulfate esters, sulfonic acid esters, amino acid esters, gluconate esters, and various amides, acetals, hemiacetals, and carbonate amide esters.

[0082] The drug or pharmaceutical composition according to the present invention can be administered orally, topically, parenterally, or mucosally (e.g., sublingually, by inhalation, or rectally) in dosage unit formulations containing a conventional non-toxic, pharmaceutically acceptable carrier. Oral administration is usually preferred. The active agent can be administered orally in the form of capsules, tablets, etc. (see Remington: The Science and Practice of Pharmacy, 20th Edition).

[0083] When administered orally in tablet or capsule form, the active drug component may be combined with non-toxic, pharmaceutically acceptable adjuvants such as binders (e.g., pre-gelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose), fillers (e.g., lactose, sucrose, glucose, mannitol, sorbitol, other reducing and non-reducing sugars, microcrystalline cellulose, calcium sulfate, or calcium hydrogen phosphate), lubricants (e.g., magnesium stearate, talc, or silica, stearic acid, sodium stearyl fumarate, glyceryl docosanate, calcium stearate, etc.), disintegrants (e.g., potato starch or sodium hydroxystarch acetate), or wetting agents (e.g., sodium lauryl sulfate), colorants and flavorings, gelatin, sweeteners, natural and synthetic gums (e.g., gum arabic, tragacanth gum, or alginates), buffer salts, carboxymethylcellulose, polyethylene glycol, and waxes. When administered orally in liquid form, the drug component can be combined with a non-toxic and pharmaceutically acceptable inert carrier (e.g., ethanol, glycerol, water), an anti-sedimentation agent (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible oils), an emulsifier (e.g., lecithin or gum arabic), a non-aqueous carrier (e.g., almond oil, ester oil, ethanol, or fractionated vegetable oil), a preservative (e.g., methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, or sorbic acid), etc. Stabilizers such as antioxidants (BHA, BHT, citronellyl propionate, sodium ascorbate, citric acid) may also be added to stabilize the dosage form.

[0084] Tablets containing the active compound can be coated by methods well known in the art. The compositions according to the present invention, comprising the compound of formula I as the active compound, can also be introduced into beads, microspheres, or microcapsules constructed from, for example, polyglycolic acid / lactic acid (PGLA). Liquid formulations for oral administration can take the form of, for example, solutions, syrups, emulsions, suspensions, or dry formulations reconstituted with water or other suitable adjuvants before use. Formulations for oral administration can be appropriately formulated to release the active compound in a controlled or delayed manner.

[0085] The term "treatment" includes the inhibition, alleviation, prevention, or elimination of one or more symptoms or side effects associated with the disease, condition, or disorder being treated.

[0086] The term "inhibition" is used in relation to a control. Those skilled in the art can easily determine an appropriate control for each experiment. For example, the reduction in response in subjects or cells treated with the compound is compared to the response in subjects or cells not treated with the compound.

[0087] The term "pharmaceutical composition" means a composition comprising the compound according to the present invention or a pharmaceutically acceptable salt thereof, and at least one selected from pharmaceutically acceptable components, including but not limited to carriers, diluents, adjuvants, excipients, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, dispersants, thermosensitive materials, temperature regulators, adhesives, stabilizers, suspension aids, etc., depending on the method of administration and the properties of the dosage form.

[0088] The terms “effective dose” or “therapeutic effective dose” mean a non-toxic amount of drug or agent sufficient to achieve the desired effect. In embodiments of the present invention, when treating a patient according to the present invention, the predetermined amount of drug to be administered depends on many factors such as the specific administration protocol, the type and severity of the disease or condition, and the specificity of the patient or host requiring treatment (e.g., body weight). However, the dose can be routinely determined by methods known in the art, depending on the specific surrounding circumstances, including the specific drug employed, the route of administration, the condition being treated, and the patient or host being treated. Typically, the dose for use in treating adults is in the range of 0.02 to 5000 mg / day, for example, about 1 to 1500 mg / day. This required dose may, for convenience, be expressed as a single dose or as divided doses administered simultaneously (or for a short period) or at appropriate intervals, for example, two, three, four or more divided doses per day. While the above dosage range is indicated, it will be understood by those skilled in the art that the specific effective dose can be appropriately adjusted according to the patient's condition and in conjunction with the physician's diagnosis.

[0089] The term "antibody-drug conjugate (ADC)" refers to a monoclonal antibody to which a biologically active small molecule drug is chemically linked, with the monoclonal antibody acting as a carrier for transporting the small molecule drug to target cells.

[0090] In this specification, the terms “reduction,” “inhibition,” “mitigation,” or “reduction” are used in relation to a control. Those skilled in the art can easily determine an appropriate control for each experiment. For example, the reduction in response in subjects or cells treated with the compound can be compared to the response in subjects or cells not treated with the compound.

[0091] Unless otherwise specified, the raw materials and apparatus used in the specific embodiments of the present invention are known products and can be obtained by purchasing commercially available products.

[0092] Preparation Example 1: General synthesis method A: Formaldehyde (1-3 eq), amine compound (1-2 eq), and dioxane were added to a reaction bottle, heated to 50-70°C with stirring, reacted for 1 hour, cooled to room temperature, 1 eq of 10-hydroxycamptothecin was added, the temperature was raised to 70-90°C, and the reaction continued for 4 hours with stirring, cooled, filtered, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (DCM:MeOH=90:1-10:1) to obtain the target compound. Removal of Boc protecting group: The Boc-protected compound obtained above was added to a reaction bottle, ethyl acetate was added to dissolve the product, stirred at 0°C, 4M HCl solution of ethyl acetate was added, stirred at room temperature for 2 hours, concentrated, and recrystallized to obtain the final compound.

[0093] General synthesis method B: 10-hydroxycamptothecin (1 eq) and hexamethylenetetramine (1.2-2.0 eq) were reacted in TFA under an argon gas atmosphere with stirring at 50-70°C for 12 hours. The reaction mixture was concentrated, H2O was added and stirred for 1 hour, the pH was adjusted to 8-9 with saturated NaHCO3 aqueous solution, extracted with ethyl acetate, the aqueous phase was acidified to 4-5 pH with 2N HCl and extracted with ethyl acetate, dried over Na2SO4, purified by silica gel column chromatography, washed with MeOH / DCM (1:50), and concentrated to obtain the 9-formylated camptothecin product.

[0094] 9-Formyl-10-hydroxycamptothecin (1 eq), an amine compound (2 eq), and methanol / dichloromethane were added to a reaction bottle and reacted at room temperature for 30 minutes. Sodium cyanoborohydride was added and the reaction was continued at room temperature with stirring for 6 hours. The mixture was filtered, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, purified by silica gel column chromatography, and eluted with DCM:MeOH = 90:1 to 10:1 to obtain the target compound.

[0095] Removal of the Boc protecting group: The same procedure as in method A above was followed, and the final compound was obtained by recrystallization.

[0096] Example 1: 9-(1-(4-aminopiperidine))methyl-10-hydroxycamptothecin(1) [ka] Method A: Paraformaldehyde (8.3 mg, 0.274 mmol), 4-Boc-aminopiperidine (27 mg, 0.137 mmol), and dioxane (10 mL) were heated to 70°C with stirring and reacted for 2 hours. After cooling to room temperature, 10-hydroxycamptothecin (25 mg, 0.07 mmol) was added, the temperature was raised to 110°C, and the mixture was reacted for 6 hours with stirring. After cooling, the mixture was filtered, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, filtered by column, and the target compound 9-(4-Boc-aminopiperidine)methyl-10-hydroxycamptothecin was obtained using an eluent DCM:MeOH ratio of 90:1 to 10:1 (37.1 mg, yield 93.7%). LC / MS (M+H): 577.11 (calculated value: 576.26).

[0097] The above 9-(4-Boc-aminopiperidine)methyl-10-hydroxycamptothecin (37.1 mg) was dissolved in ethyl acetate (2 mL), stirred at 0°C, and 0.5 mL of ethyl acetate solution in 4 M HCl was added. The mixture was stirred at room temperature for 2 hours. The solvent was concentrated, and the mixture was recrystallized from ethyl acetate / petroleum ether to obtain the red solid compound 9-(1-(4-aminopiperidine))methyl-10-hydroxycamptothecin as the product (30.1 mg, yield 90.9%). 1 H NMR(500MHz,DMSO-d6)δ8.84(s,1H),8.37-8.15(m,3H),8.10(d,J=9.2Hz, 1H),7.61(d,J=9.2Hz,1H),7.28(s,1H),6.35(s,3H),5.42(s,2H),5.26(s, 2H),4.48(s,2H),3.28(d,J=11.4Hz,1H),2.95(s,2H),2.03(d,J=12.9Hz,2 H),1.87(dt,J=16.2,7.1Hz,2H),1.83-1.68(m,2H),0.88(t,J=7.3Hz,3H). LC / MS(M+H): 476.99 (Calculated value: 476.21).

[0098] Example 2: 9-(1-piperazine)methyl-10-hydroxycamptothecin(2) [ka] Paraformaldehyde, 1-Boc piperazine, dioxane, and 10-hydroxycamptothecin were prepared according to Method A to obtain compound 9-(4-Boc-piperazine-1-)methyl-10-hydroxycamptothecin (yield: 92.2%). LC / MS(M+H): 563.05 (calculated value: 562.24).

[0099] The above 9-(4-Boc-piperazine-1-)methyl-10-hydroxycamptothecin (35.6 mg) was dissolved in ethyl acetate (2 mL), and while stirring at 0°C, 4M ethyl acetate solution (0.5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated, and recrystallized from ethyl acetate / petroleum ether to obtain the solid compound 9-(1-piperazine)methyl-10-hydroxycamptothecin as the product (26.8 mg, yield 91.2%). 1 HNMR(500MHz,DMSO-d6)δ11.59(s,1H),9.93(s,2H),9.10(s,1H),8.13(dt,J=9.2,3.0Hz,1H),7.76(d,J=9.2Hz,1H),7.26(t,J=1.8Hz,1H) ,5.39(s,2H),5.19-5.15(m,2H),4.80(s,2H),3.73(s,2H),1.97(s,1H),1.90(s,2H),1.86(dd,J=10.7,7.1Hz,2H),0.87(t,J=7.3Hz,3H). LC / MS (M+H): 462.97 (calculated value: 462.19).

[0100] Example 3: 9-(1-(4-aminooxypiperidine))methyl-10-hydroxycamptothecin(3) [ka] Preparation of 2-(piperidine-4-oxy)isodihydroindole-1,3-dione: Triphenylphosphine (1.93 g, 7.34 mmol) and N-hydroxyphthalimide (1.20 g, 7.34 mmol) were dissolved in THF (10 mL), stirred at 0°C, and 1-Boc-4-hydroxypiperidine (0.500 g, 4.90 mmol) and diethyl azodicarboxylate (3.34 mL, 7.34 mmol) were added. The mixture was slowly heated to room temperature, stirred overnight, and the solvent was concentrated under reduced pressure. The solution was purified by silica gel column chromatography and eluted with petroleum ether / ethyl acetate (90 / 10~50 / 50) to obtain the product 2-(1-Boc-piperidine-4-oxy)isodihydroindole-1,3-dione (1.8 g, yield 72%). LC / MS (M+H): 347.02 (calculated value: 346.15).

[0101] 2-(1-Boc-piperidine-4-oxy)isodihydroindole-1,3-dione (1.8 g, 5.2 mmol) was dissolved in ethyl acetate (5 mL), and 2 mL of 4 M HCl ethyl acetate solution was added. The mixture was stirred at room temperature for 1 hour, and then concentrated to obtain 2-(piperidine-4-oxy)isodihydroindole-1,3-dione hydrochloride (1.31 g, yield 89%). LC / MS (M+H): 247.07 (calculated value: 246.10).

[0102] Paraformaldehyde, 2-(piperidine-4-oxy)isodihydroindole-1,3-dione hydrochloride, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(2-(piperidine-4-oxy)isodihydroindole-1,3-dione-1-)methyl-10-hydroxycamptothecin (yield 66.5%). LC / MS(M+H): 623.05 (calculated value: 622.21).

[0103] 9-(2-(piperidine-4-oxy)isodihydroindole-1,3-dione-1-)methyl-10-hydroxycamptothecin was added to a reaction bottle, ethanol and 85% hydrazine hydrate (1.5 eq) were added, the mixture was stirred at room temperature for 1 hour, concentrated, converted to hydrochloride with 4M HCl-containing ethyl acetate, and recrystallized from ethyl acetate / petroleum ether to obtain the red solid compound 9-(1-(4-aminooxypiperidine))methyl-10-hydroxycamptothecin (yield: 85.6%). 1 H NMR(600MHz,DMSO-d6)δ11.50(s,1H),11.01(s,2H),10.19(s,1H),9.07(s,1H),8.16 (d,J=9.2Hz,1H),7.74(dd,J=9.2,6.3Hz,1H),7.28(s,1H),6.55(s,1H),5.42(s,2H), 5.24(s,2H),4.74(s,2H),4.41(d,J=43.2Hz,1H),3.30(s,2H),3.06(dd,J=7.3,4.8H z,1H),2.19(s,3H),1.88(dp,J=21.6,7.2Hz,2H),1.55(s,1H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 493.00 (calculated value: 492.20).

[0104] Example 4: 9-(O-ethylhydroxyamine)methyl-10-hydroxycamptothecin (4) [ka] 9-Formyl-10-hydroxycamptothecin, O-ethylhydroxyamine hydrochloride, methanol, and cyano were synthesized according to Method B to obtain compound 9-(O-ethylhydroxyamine)methyl-10-hydroxycamptothecin (yield 56.3%). 1H NMR(600MHz,DMSO-d6)δ11.37(s,1H),8.87(s,1H),8.69(s,2H),8.15(d,J=9.1Hz,1H),7.64(d,J=9.2Hz,1H),7.29(s,1H),6.52(s,1H),5.43( s,2H),5.27(s,2H),4.57(s,2H),3.13(q,J=7.6,7.1Hz,2H),1.87(ddt,J=21.2,14.5,7.2Hz,2H),1.27(t,J=7.2Hz,3H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 438.08 (calculated value: 437.16).

[0105] Example 5: 9-(1-(4-methoxypiperidine))methyl-10-hydroxycamptothecin (5) [ka] Paraformaldehyde, 4-hydroxymethylpiperidine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(4-methoxypiperidine-1-)methyl-10-hydroxycamptothecin (yield 81.5%). 1 H NMR(500MHz,DMSO-d6)δ11.39(s,1H),9.30(s,1H),8.94(s,1H),8.19(d,J=9.2Hz,1H),7.64(d,J=9.2Hz,1H),7.29(s,1H),6.52 (s,1H),5.43(s,2H),5.29(s,2H),4.72(s,2H),3.36(s,5H),2.13(s,1H),2.04-1.81(m,4H),1.60(s,1H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 492.06 (calculated value: 491.21).

[0106] Example 6: 9-(1-(4-cyclopropylcarbonylpiperazine))methyl-10-hydroxycamptothecin(6) [ka] Using paraformaldehyde, 1-cyclopropylcarbonylpiperazine, dioxane, and 10-hydroxycamptothecin, compound 9-(4-cyclopropylcarbonylpiperazine-1-)methyl-10-hydroxycamptothecin was obtained according to Method A (yield 91.1%). 1 H NMR(500MHz,DMSO-d6)δ8.74(d,J=9.0Hz,1H),7.96(t,J=8.9Hz,1H),7.45(dd,J=9.5,4.6Hz,1H), 7.25(d,J=3.3Hz,1H),5.41(s,2H),5.20(d,J=11.3Hz,2H),4.02(d,J=3.8Hz,2H),3.53-3.45(m,4 H),3.43-3.39(m,1H),3.37(t,J=6.5Hz,1H),2.58(s,2H),2.02-1.92(m,1H),1.87(dt,J=16.5,7. 2Hz,2H), 1.46(dd,J=8.5,6.0Hz,1H),1.35-1.27(m,1H),0.89(t,J=7.5Hz,3H),0.82-0.60(m,4H). LC / MS (M+H): 531.04 (calculated value: 530.22).

[0107] Example 7: 9-(1-(4-acetylpiperazine))methyl-10-hydroxycamptothecin (7) [ka] Paraformaldehyde, 1-acetylpiperazine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(4-acetylpiperazine-1-)methyl-10-hydroxycamptothecin (yield 79.6%). 1H NMR(500MHz,DMSO-d6)δ11.46(s,1H),9.86(s,1H),8.93(s,1H),8.18(d,J=9.2Hz,1H),7.64(d,J=9.2Hz,1H),7.29(s,1H) ,6.52(s,1H),5.43(s,2H),5.29(s,2H),4.73(s,2H),3.47(s,7H),2.05(s,3H),1.96-1.78(m,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 505.07 (calculated value: 504.20).

[0108] Example 8: 9-(1-(4-carbamoylpiperazine))methyl-10-hydroxycamptothecin(8) [ka] Using paraformaldehyde, piperidine-1-carboxamide, dioxane, and 10-hydroxycamptothecin, compound 9-(4-carbamoylpiperazine-1-)methyl-10-hydroxycamptothecin was obtained according to Method A (yield 84.2%). 1 H NMR(500MHz,DMSO-d6)δ8.71-8.61(m,1H),7.98-7.88(m,1H),7.41-7.33(m,1H),7.31-7.25(m,1H),7.25-7. 21(m,1H),6.77(s,1H),5.49-5.32(m,2H),5.27-5.13(m,2H),3.47(tdd,J=9.7,6.4,4.2Hz,3H),3.40(t,J=5. 3Hz,1H),3.36(td,J=6.6,1.9Hz,1H),3.06-2.93(m,2H),2.29-2.13(m,2H),1.86(dt,J=15.0,7.2Hz,2H),1. 76-1.70(m,1H),1.59(td,J=12.7,12.3,3.6Hz,2H),1.51-1.40(m,1H),1.32-1.26(m,1H),0.89-0.85(m,3H). LC / MS (M+H): 505.01 (calculated value: 504.20).

[0109] Example 9: 9-(1-(4-methoxycarbonylpiperazine))methyl-10-hydroxycamptothecin(9) [ka] Paraformaldehyde, methyl piperazine-1-carboxylate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(4-methoxycarbonylpiperazine-1-)methyl-10-hydroxycamptothecin (yield 87.5%). 1 H NMR(500MHz,DMSO-d6)δ11.42(s,1H),9.73(s,1H),8.92(s,1H),8.17(d,J=9.0Hz,1H),7.63(d,J=9.0Hz,1H),7.29(s,1H),6.52(s,1 H),5.43(s,2H),5.29(s,2H),4.71(s,2H),4.03(s,2H),3.64(s,3H),3.27(s,5H),1.87(m,J=18.2,7.2Hz,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 521.07 (calculated value: 520.20).

[0110] Example 10: 9-(N-methyl-N-isopropylamino)methyl-10-hydroxycamptothecin (10) [ka] Using paraformaldehyde, N-methylpropan-2-amine, dioxane, and 10-hydroxycamptothecin, compound 9-(N-methylisopropylamino)methyl-10-hydroxycamptothecin was obtained according to Method A (yield 87.5%). 1H NMR(500MHz,DMSO-d6)δ8.65(s,1H),7.90(d,J=9.1Hz,1H),7.30(d,J=9.1Hz,1H),7.23(s,1H),5.40(d,J=2.2Hz,2H),5.17(s,2 H),4.19(s,2H),3.08(q,J=6.6Hz,1H),2.24(s,3H),1.87(dq,J=14.8,7.2Hz,2H),1.14(d,J=6.6Hz,6H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 450.04 (calculated value: 449.20).

[0111] Example 11: 9-(N-4-aminobutyl-N-methylamino)methyl-10-hydroxycamptothecin (11) [ka] Formaldehyde, tert-butyl(4-(methylamino)butyl)-1-carbamate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-4-Bocaminobutyl-N-methylamino)methyl-10-hydroxycamptothecin (yield 82.7%). LC / MS(M+H): 579.07 (calculated value: 578.27).

[0112] 9-(N-4-Bocaminobutyl-N-methylamino)methyl-10-hydroxycamptothecin was dissolved in ethyl acetate, stirred at 0°C, a 4M HCl solution of ethyl acetate was added, stirred at room temperature for 2 hours, concentrated, and recrystallized to obtain the red solid compound 9-(N-4-aminobutyl-N-methylamino)methyl-10-hydroxycamptothecin (yield 88.3%). 1H NMR(600MHz,DMSO-d6)8.93-8.84(m,1H),8.73(s,1H),8.19(ddd,J=19.2,9.2,3.4Hz,1 H),7.90-7.71(m,2H),7.64(td,J=8.9,8.5,3.7Hz,1H),7.30(s,1H),6.53(s,1H),5.43( s,2H),5.29(s,2H),4.86-4.63(m,1H),2.90-2.79(m,1H),2.71(d,J=32.2Hz,2H),1.87 (m,J=21.4,14.3,7.2Hz,2H),1.59(d,J=7.9Hz,1H),1.23(s,1H),0.88(t,J=7.3Hz,3H). LC / MS (M+H): 479.04 (calculated value: 478.22).

[0113] Example 12: 9-(N-methylhydroxyethylamino)methyl-10-hydroxycamptothecin (12) [ka] Paraformaldehyde, N-methylethanolamine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methylhydroxyethylamino)methyl-10-hydroxycamptothecin (yield 90.7%). 1 H NMR(500MHz,DMSO-d6)δ11.44(s,1H),9.20(s,1H),8.88(s,1H),8.19(d,J=9.2Hz,1H),7.64(dd,J=9.2,1.9Hz,1H),7.30(s,1H),6.52(s ,1H),5.43(s,3H),5.28(s,2H),4.87(s,1H),4.74(s,1H),3.86(s,2H),2.80(s,3H),1.87(dp,J=21.4,7.2Hz,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 452.00 (calculated value: 451.17).

[0114] Example 13: 9-(N-methyl-N-aminoethylamino)methyl-10-hydroxycamptothecin (13) [ka] Method B: 9-Formyl-10-hydroxycamptothecin (25 mg, 0.064 mmol), tert-butyl(2-(methylamino)ethyl)carbamate (10.2 mg, 0.07 mmol), and methanol (3 mL) were added to a reaction bottle and reacted at 20°C for 1 hour with stirring. After cooling to 0°C, sodium cyanoborohydride (4.4 mg, 0.07 mmol) was added and the reaction continued at 20°C for 2 hours with stirring. The mixture was concentrated and purified by silica gel column chromatography (DCM / MeOH: 90:1~10:1) to obtain compound 9-(N-methyl-N-Bocaminoethylamino)methyl-10-hydroxycamptothecin (34.1 mg, yield 95.7%). LC / MS (M+H): 551.21 (calculated value: 550.24).

[0115] 9-(N-methyl-N-Bocaminoethylamino)methyl-10-hydroxycamptothecin (34.1 mg) was dissolved in ethyl acetate (2 mL), stirred at 0°C, and ethyl acetate solution in 4 M HCl (0.5 mL) was added. The mixture was stirred at room temperature for 2 hours, concentrated, and recrystallized from ethyl acetate / petroleum ether to obtain the pale red solid compound 9-(N-methyl-N-aminoethylamino)methyl-10-hydroxycamptothecin (27.6 mg, yield 87.7%). 1 H NMR(500MHz,DMSO-d6)δ11.65(s,1H),9.77(s,1H),8.89(s,1H),8.17(d,J=9.2Hz,1H),8.14-7.78(m,2H),7.65(d,J=9.1Hz,1H),7 .30(s,1H),5.43(s,2H),5.28(s,2H),4.68(s,2H),3.24(s,5H),2.77(s,2H),1.88(dp,J=14.3,7.1Hz,2H),0.89(t,J=7.2Hz,3H). LC / MS (M+H): 451.19 (calculated value: 450.19).

[0116] Example 14: 9-(N-methyltetrahydropyran-4-amine)methyl-10-hydroxycamptothecin (14) [ka] Paraformaldehyde, N-methyltetrahydro-2H-pyran-4-amine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methyltetrahydropyran-4-amino)methyl-10-hydroxycamptothecin (yield 88.7%). 1 H NMR(500MHz,DMSO-d6)δ11.56(s,1H),9.22(s,1H),8.87(s,1H),8.20(dd,J=9.2,2.3H z,1H),7.66(dd,J=9.2,2.0Hz,1H),7.29(d,J=1.1Hz,1H),6.53(s,1H),5.43(s,2H),5 .35-5.19(m,2H),4.73(d,J=120.2Hz,2H),4.15-3.98(m,2H),3.71(s,1H),2.67(s,3H) ),2.19-2.06(m,2H),1.88(ddq,J=21.3,14.1,7.1,6.5Hz,4H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 492.12 (calculated value: 491.21).

[0117] Example 15: 9-(trans-4-hydroxycyclohexylamine)methyl-10-hydroxycamptothecin (15) [ka] Paraformaldehyde, trans-4-aminocyclohexane-1-ol, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(4-hydroxycyclohexylamine)methyl-10-hydroxycamptothecin (yield 76.9%). 1H NMR(500MHz,DMSO-d6)δ8.64-8.54(m,1H),7.94-7.82(m,1H),7.34-7.10(m,2H),5.39(s,2H),5.27-5.11(m,2H),4.36(s,2H),4.30-4.12(m ,4H),3.41(dp,J=8.5,4.2Hz,2H),1.98(d,J=10.0Hz,2H),1.85(tq,J=14.5,8.4,7.8Hz,4H),1.19(q,J=12.5Hz,4H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 492.09 (calculated value: 491.21).

[0118] Example 16: 9-(N-methyl-trans-4-hydroxycyclohexylamine)methyl-10-hydroxycamptothecin (16) [ka] Paraformaldehyde, trans-4-(methylamino)cyclohexane-1-ol, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methyl-4-hydroxycyclohexylamine)methyl-10-hydroxycamptothecin (yield 82%). 1 H NMR(600MHz,DMSO-d6)δ11.63(s,1H),9.15(s,1H),8.86(s,1H),8.19(d,J=9.1Hz,1H ),7.66(d,J=9.2Hz,1H),7.28(s,1H),6.55(s,1H),5.43(s,2H),5.26(s,2H),4.77(d, J=12.7Hz,1H),4.72-4.51(m,1H),3.50-3.39(m,2H),2.65(s,3H),2.26-2.08(m,2H) ,2.00(s,2H),1.93-1.62(m,4H),1.29(td,J=13.2,6.6Hz,2H),0.89(t,J=7.4Hz,3H). LC / MS (M+H): 506.02 (calculated value: 505.22).

[0119] Example 17: 9-(N-methyl-N-cyclopropylamine)methyl-10-hydroxycamptothecin (17) [ka] Using paraformaldehyde, N-methylcyclopropylamine, dioxane, and 10-hydroxycamptothecin, compound 9-(N-methyl-N-cyclopropylamine)methyl-10-hydroxycamptothecin was obtained according to Method A (yield 83.9%). 1 H NMR(500MHz,DMSO-d6)δ8.55(s,1H),7.87(d,J=9.1Hz,1H),7.35(d,J=9.2Hz,1H),7.17(s,1H),5.3 4(d,J=2.7Hz,2H),5.13(s,2H),4.09(s,2H),3.42(tdd,J=9.9,6.5,4.2Hz,3H),3.35(t,J=5.3Hz,1 H),3.30(t,J=6.6Hz,1H),2.19(s,3H),1.86(ddt,J=9.8,7.1,3.1Hz,1H),1.79(p,J=7.1Hz,2H),1. 39(dd,J=8.4,6.2Hz,1H),1.29-1.19(m,1H),0.87-0.81(m,3H);LC / MS(M+H):448.02(calculated value:447.18).

[0120] Example 18: 9-(morpholine-4-amine)methyl-10-hydroxycamptothecin (18) [ka] 9-Formyl-10-hydroxycamptothecin, 4-aminomorpholine, methanol, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(morpholine-4-amine)methyl-10-hydroxycamptothecin (yield 81.0%). LC / MS(M+H): 479.08 (calculated value: 478.19).

[0121] Example 19: 9-(N-methylethylamine)methyl-10-hydroxycamptothecin (19) [ka] Paraformaldehyde, N-methylethylamine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methyl-N-ethylamine)methyl-10-hydroxycamptothecin (yield 77.3%). 1 H NMR(500MHz,DMSO-d6)δ11.45(s,1H),8.88(s,1H),8.18(d,J=9.2Hz,1H),7.64(d,J=9.2Hz,1H),7.29(s,1H),6.52(s,1H),5.43(s,2H), 5.27(s,2H),4.70(s,2H),3.28(m,J=7.0Hz,2H),2.73(s,3H),1.87(d,J=21.5,7.2Hz,2H),1.33(t,J=7.2Hz,3H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 436.02 (calculated value: 435.18).

[0122] Example 20: 9-(ethylamine)methyl-10-hydroxycamptothecin (20) [ka] Paraformaldehyde, ethylamine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-ethylamine)methyl-10-hydroxycamptothecin (yield 84.0%). 1 H NMR(500MHz,DMSO-d6)δ8.74(s,1H),7.98(d,J=9.1Hz,1H),7.47(t,J=9.2Hz,1H),7.25(s,1H),5.41(s,2H),5.23(s,2H) ),4.42(s,2H),2.85(dq,J=27.0,7.3Hz,4H),1.87(dt,J=15.5,8.2Hz,3H),1.18(d,J=7.5Hz,3H),0.88(t,J=7.4Hz,3H). LC / MS (M+H): 422.06 (calculated value: 421.16).

[0123] Example 21: 9-(isopropylamine)methyl-10-hydroxycamptothecin (21) [ka] Paraformaldehyde, isopropylamine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(isopropylamine)methyl-10-hydroxycamptothecin (yield 79.7%). 1 H NMR(500MHz,DMSO-d6)δ11.45(s,1H),8.83(s,1H),8.60(s,2H),8.14(d,J=9.2Hz,1H),7.64(d,J=9.2Hz,1H),7.28(s,1H),6.52(s ,1H),5.42(s,2H),5.25(s,2H),4.55(d,J=6.2Hz,2H),1.87(dp,J=18.1,7.1Hz,2H),1.38(d,J=6.5Hz,6H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 436.02 (calculated value: 435.18).

[0124] Example 22: 3-(N-morpholine)-1,3-oxazine[5,6],[9,10]camptothecin(22) [ka] A 40% aqueous formaldehyde solution, morpholine-4-amine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 3-(N-morpholino)-1,3-oxazino[5,6],[9,10]camptothecin (yield 64.3%). LC / MS(M+H): 491.07 (calculated value: 490.19).

[0125] Example 23: 9-(1-methyl-2-Boc hydrazine)methyl-10-hydroxycamptothecin (23) [ka] Paraformaldehyde, tert-butyl 2-methylhydrazine-1-carboxylate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 1-methyl-2-Boc hydrazine methyl-10-hydroxycamptothecin (yield 90.7%). 1H NMR(500MHz,DMSO-d6)δ8.75(s,1H),8.01(d,J=9.1Hz,1H),7.50(d,J=9.1Hz,1H),7.26(s,1H),6.44(d,J=56.6Hz ,2H),5.41(s,3H),5.24(s,2H),4.93(s,2H),2.41(s,3H),1.93-1.84(m,2H),1.48(s,9H),0.89(d,J=7.3Hz,3H). LC / MS (M+H): 523.08 (calculated value: 522.21).

[0126] Example 24: 9-(1-methylhydrazine)methyl-10-hydroxycamptothecin (24) [ka] 1-Methyl-2-Boc hydrazine methyl-10-hydroxycamptothecin (compound 23) was dissolved in ethyl acetate (2 mL), stirred at 0°C, and 0.5 mL of ethyl acetate solution in 4 M HCl was added. The mixture was stirred at room temperature for 2 hours, concentrated, and recrystallized from petroleum ether / ethyl acetate to obtain solid 1-methylhydrazine methyl-10-hydroxycamptothecin as the product (yield 89.5%). 1 H NMR(600MHz,DMSO-d6)δ10.86(s,1H),9.97(s,2H),8.72(s,1H),8.05(d,J=9.2Hz,1H),7.57(d,J=9.2Hz,1H),7.27(s,1H),6.51(s,1 H),5.86(s,1H),5.42(s,2H),5.26(s,2H),4.49(s,2H),2.75(d,J=26.1Hz,3H),1.87(dq,J=10.7,7.1Hz,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 423.08 (calculated value: 422.16).

[0127] Example 25: 9-(1-methyl-3-aminopropylamine)methyl-10-hydroxycamptothecin (25) [ka] 9-Formyl-10-hydroxycamptothecin, tert-butyl(3-(methylamino)propyl)carbamate, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(1-methyl-3-Boc-aminopropylamine)methyl-10-hydroxycamptothecin (yield 90.6%). LC / MS(M+H): 565.17 (calculated value: 564.26).

[0128] The above Boc group protecting product was dissolved in ethyl acetate (2 mL), stirred at 0°C, and 0.5 mL of ethyl acetate solution in 4 M HCl was added. The mixture was stirred at room temperature for 2 hours, the solvent was concentrated, and the mixture was recrystallized from ethyl acetate / petroleum ether to obtain the solid product 9-(1-methyl-3-aminopropylamine)methyl-10-hydroxycamptothecin (yield 85.5%). 1 H NMR(500MHz,DMSO-d6)δ8.55(s,1H),7.87(d,J=9.1Hz,1H),7.35(d,J=9.2Hz,1H),7.17( s,1H),5.34(d,J=2.7Hz,2H),5.13(s,2H),4.09(s,2H),3.42(m,J=9.9,6.5,4.2Hz,3H),3 .35(t,J=5.3Hz,1H),3.30(t,J=6.6Hz,1H),2.19(s,3H),1.86(m,J=9.8,7.1,3.1Hz,1H), 1.79(p,J=7.1Hz,2H),1.39(dd,J=8.4,6.2Hz,1H),1.29-1.07(m,2H),0.87-0.81(m,3H). LC / MS (M+H): 465.09 (calculated value: 464.21).

[0129] Example 26: 9-(piperidine-1-amine)methyl-10-hydroxycamptothecin (26) [ka] 9-Formyl-10-hydroxycamptothecin, 1-aminopiperidine, methanol, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(piperidine-1-amine)methyl-10-hydroxycamptothecin (yield 87.6%). LC / MS(M+H): 477.20 (calculated value: 476.21).

[0130] Example 27: 9-(N-methyl(tetrahydropyranmethyl)amine)methyl-10-hydroxycamptothecin (27) [ka] Paraformaldehyde, N-methyl-1-(tetrahydro-2H-pyran-4-yl)methylamine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methyl(tetrahydropyranmethyl)amine)methyl-10-hydroxycamptothecin (yield 87%). 1 H NMR(500MHz,DMSO-d6)δ11.75(s,1H),8.88(d,J=3.3Hz,1H),8.18(dd,J=9.3,3.0Hz,1H),7.65(dd,J=9.4,3. 6Hz,1H),7.29(d,J=2.9Hz,1H),6.54(s,1H),5.42(d,J=3.1Hz,2H),5.26(s,2H),4.76(s,2H),3.88(d,J=11.3 Hz,2H),3.34(d,J=11.6Hz,2H),3.20(s,2H),2.73(d,J=3.5Hz,3H),2.25(ddd,J=11.4,7.6,3.9Hz,1H),1.88 (ddd,J=16.5,11.9,7.2Hz,2H),1.76(d,J=12.8Hz,2H),1.26(q,J=12.3Hz,2H),1.08-0.78(d,J=12.8Hz,3H). LC / MS (M+H): 506.09 (calculated value: 505.22).

[0131] Example 28: 9-(N-methyl(piperidine-4-methyl)amine)methyl-10-hydroxycamptothecin (28) [ka] Formaldehyde, tert-butyl 4-((methylamino)methyl)piperidine-1-carboxylate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methyl(1-Boc-piperidine-4-methyl)amine)methyl-10-hydroxycamptothecin (yield 93.3%). LC / MS(M+H): 605.14 (calculated value: 604.29). The Boc-protected intermediate was dissolved in ethyl acetate (2 mL), stirred at 0°C, 0.5 mL of ethyl acetate solution in 4 M HCl was added, stirred at room temperature for 2 hours, concentrated, and recrystallized to obtain the solid product 9-(N-methyl(piperidine-4-methyl)amine)methyl-10-hydroxycamptothecin (yield 88.5%). 1 H NMR(500MHz,DMSO-d6)δ11.65(s,1H),8.89(s,1H),8.19(dd,J=9.7,3.8Hz,1H),7.64(dd,J=9.2,1.7Hz,1H) ,7.29(d,J=2.1Hz,1H),6.52(s,1H),5.42(s,2H),5.28(d,J=5.8Hz,2H),4.76(s,2H),3.87(dt,J=10.9,3.4 Hz,2H),3.34((td,J=11.82.1Hz,2H),3.19(s,2H),2.73(s,3H),2.24(ddd,J=11.4,7.6,4.1Hz,1H),1.94-1 .82(m,2H),1.75(s,2H),1.26(d,J=14.3Hz,2H),0.88(t,J=7.3Hz,3H): 505.14 (calculated value: 504.24).

[0132] Example 29: 9-(3-(Undeca-5,5-spirocyclo-3,9-diazan))methyl-10-hydroxycamptothecin (29) [ka] Formaldehyde, tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(3-(9-Boc-undeca-5,5-spiro-3,9-diazan))methyl-10-hydroxycamptothecin (yield 92.9%). LC / MS(M+H): 631.18 (calculated value: 630.31).

[0133] The above Boc- intermediate was dissolved in ethyl acetate (2 mL), stirred at 0°C, and 0.5 mL of ethyl acetate solution in 4 M HCl was added. The mixture was stirred at room temperature for 2 hours, concentrated, and recrystallized from ethyl acetate / petroleum ether to obtain the solid product 9-(3-(undeca-5,5-spiro-3,9-diazan))methyl-10-hydroxycamptothecin (yield 91.1%). 1 H NMR(500MHz,DMSO-d6)δ11.66(s,1H),9.59(s,1H),8.92(s,1H),8.73(s,2H),8.15(d,J=9.2Hz,1H),7.67(d,J=9.3Hz,1H),7.28(s,1H),6.52( s,1H),5.42(s,2H),5.23(s,2H),4.75(s,2H),3.69-3.25(m,8H),1.87 (dq,J=14.3,7.5Hz,4H),1.58(d,J=82.2Hz,4H),0.90(t,J=7.3Hz,3H). LC / MS (M+H): 531.18 (calculated value: 530.25).

[0134] Example 30: 9-(piperidine-4-ethylamine)methyl-10-hydroxycamptothecin (30) [ka] Formaldehyde, tert-butyl 4-(2-aminoethyl)piperidine-1-carboxylate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(1-Boc-piperidine-4-ethylamine)methyl-10-hydroxycamptothecin (yield 91.1%). LC / MS(M+H): 605.27 (calculated value: 604.29).

[0135] The above Boc- intermediate was dissolved in ethyl acetate (2 mL), stirred at 0°C, and 0.5 mL of ethyl acetate solution in 4 M HCl was added. The mixture was stirred at room temperature for 2 hours, concentrated, and recrystallized from ethyl acetate / petroleum ether to obtain the solid compound 9-(piperidine-4-ethylamine)methyl-10-hydroxycamptothecin (yield 94.5%). 1 H NMR(600MHz,DMSO-d6)δ11.62(s,1H),8.86(d,J=17.7Hz,3H),8.85-8.43(m,2H),8.16(d,J=9.2Hz,1H),7.67(d,J=9.2Hz,1H),7.29(s,1H),6.53 (s,1H),5.43(s,2H),5.27(s,2H),4.65(s,2H),3.20(t,J=6.2Hz,6H),2.80-2.57(m,6H),1.88(dhept,J=21.4,7.3Hz,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 504.87 (calculated value: 504.24).

[0136] Example 31: 9-(L-proline tert-butyl ester)methyl-10-hydroxycamptothecin (31) [ka] Paraformaldehyde, L-proline tert-butyl ester, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(L-proline tert-butyl ester)methyl-10-hydroxycamptothecin (yield 90.2%). 1H NMR(600MHz,DMSO-d6)δ11.55(s,1H),10.08(s,1H),8.89(s,1H),8.15(d,J=9.0Hz ,1H),7.62(d,J=9.2Hz,1H),7.29(s,1H),6.54(s,1H),5.43(s,2H),5.29(s,2H),4. 87(s,2H),4.49(d,J=43.8Hz,1H),3.19(s,2H),2.46(s,1H),2.04(d,J=10.0Hz,1H) ,1.88(ddt,J=27.3,14.2,7.4Hz,4H),1.35(d,J=4.2Hz,9H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 548.17 (calculated value: 547.23).

[0137] Example 32: 9-(L-proline)methyl-10-hydroxycamptothecin (32) [ka] 9-(L-proline tert-butyl ester)methyl-10-hydroxycamptothecin (compound from Example 31) was taken, dissolved in dichloromethane (2 mL), stirred at 0°C, trifluoroacetic acid (2 mL) was added, stirred at room temperature for 2 hours, concentrated, and recrystallized from ethyl acetate / petroleum ether to obtain the solid product 9-(L-proline)methyl-10-hydroxycamptothecin (yield 81.4%). 1 H NMR(500MHz,DMSO-d6)δ8.92(s,1H),8.17(d,J=9.2Hz,1H),7.62(d,J=9.3Hz,1H),7.3 0(d,J=1.5Hz,1H),6.52(s,1H),5.43(s,2H),5.36-5.20(m,2H),4.92(d,J=13.6Hz,1H ),4.82(d,J=13.7Hz,1H),4.49(t,J=8.8Hz,1H),3.33(q,J=10.0,9.4Hz,3H),2.02(tq ,J=12.6,4.3,3.2Hz,2H),1.87(ddd,J=16.6,14.1,7.5Hz,3H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 491.99 (calculated value: 491.17).

[0138] Example 33: 9-(1-piperidine-4-carboxylate methyl ester)methyl-10-hydroxycamptothecin (33) [ka] Paraformaldehyde, methylpiperidine-4-carboxylate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(1-piperidine-4-carboxylate methyl ester)methyl-10-hydroxycamptothecin (yield 91.3%). 1 H NMR(500MHz,DMSO-d6)δ11.54(s,1H),9.56(s,1H),8.92(d,J=3.7Hz,1H),8.17(t,J=8.5H z,1H),7.65(d,J=9.2Hz,1H),7.33-7.26(m,1H),5.43(s,2H),5.26(d,J=11.0Hz,2H),4.7 2(d,J=13.5Hz,2H),3.72(s,2H),3.62(s,3H),3.24(d,J=17.2Hz,2H),2.70(ddd,J=12.3, 8.6,3.7Hz,1H),2.09-1.96(m,2H),1.86(qt,J=18.5,10.0Hz,4H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 520.05 (calculated value: 519.20).

[0139] Example 34: 9-(1-piperidine-4-carboxylic acid)methyl-10-hydroxycamptothecin (34) [ka] Paraformaldehyde, piperidine-4-carboxylic acid, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(1-piperidine-4-carboxylic acid)methyl-10-hydroxycamptothecin (yield 87.5%). 1H NMR(500MHz,DMSO-d6)δ12.53(s,1H),11.40(s,1H),9.34(s,1H),8.92(s,1H) ,8.18(d,J=9.2Hz,1H),7.64(d,J=9.2Hz,1H),7.29(s,1H),6.52(s,1H),5.43( s,2H),5.28(s,2H),4.70(s,2H),3.55(s,2H),3.23(s,2H),2.56(s,1H),2.07- 1.98(m,2H),1.87(dp,J=18.3,7.1Hz,2H),1.78(s,1H),0.89(t,J=7.3Hz,3H). LC / MS(M+H): 506.02 (Calculated value: 505.18).

[0140] Example 35: 9-(L-alanine tert-butyl ester)methyl-10-hydroxycamptothecin (35) [ka] Paraformaldehyde, tert-butylmethyl-L-alanine hydrochloride, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(L-alanine tert-butyl ester)methyl-10-hydroxycamptothecin (yield 74.8%). 1 H NMR(600MHz,DMSO-d6)δ8.85(s,1H),8.15(d,J=9.1Hz,1H),7.61(d,J=9.2Hz,1H),7.29(s,1H),6.52(s,1H),5.42(s,2H),5.26(d,J=3.6Hz,2H),4. 69(d,J=36.8Hz,2H),4.27(s,1H),2.69(s,3H),1.87(dq,J=11.2,7.0Hz, 2H), 1.57(d,J=7.0Hz,2H),1.49(d,J=20.5Hz,9H),0.89(t,J=7.3Hz,3H). LC / MS(M+H): 522.07 (Calculated value: 521.22).

[0141] Example 36: 9-(N-methyl-L-alanine)methyl-10-hydroxycamptothecin (36) [ka] Paraformaldehyde, N-methyl-L-alanine hydrochloride, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(N-methyl-L-alanine)methyl-10-hydroxycamptothecin (yield 81.4%). 1 H NMR(600MHz,DMSO-d6)δ8.92(s,1H),8.16(d,J=9.1Hz,1H),7.61(d,J=9.2Hz,1H),7.29(s,1H),6.52(s,1H),5.42(s,2H),5.26(d,J=4.4H) z,2H),4.83-4.66(m,2H),4.35(d,J=9.1Hz,1H),2.72(s,2H),1.87(dp,J=21.1,7.1Hz,2H),1.63(d,J=7.2Hz,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 479.92 (calculated value: 479.17).

[0142] Example 37: 9-(tetrahydropyran-4-amine)methyl-10-hydroxycamptothecin (37) [ka] Paraformaldehyde, 4-aminotetrahydropyran, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(tetrahydropyran-4-amine)methyl-10-hydroxycamptothecin (yield 90.8%). 1H NMR(500MHz,DMSO-d6)δ11.31(s,1H),8.85(s,1H),8.72(s,2H),8.17(dt,J=9.3,2. 4Hz,1H),7.64(dt,J=9.2,1.8Hz,1H),7.29(d,J=1.4Hz,1H),6.52(s,1H),5.43(s,2H) ),5.29(d,J=2.7Hz,2H),4.61(s,2H),3.98(dd,J=10.9,4.4Hz,2H),2.13(dd,J=11.5 ,3.4Hz,2H),1.87(dq,J=18.3,7.0Hz,2H),1.76-1.62(m,2H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 478.03 (calculated value: 477.19).

[0143] Example 38: 9-(4-Boc-piperazine-1-amine)methyl-10-hydroxycamptothecin (38) [ka] 9-Formyl-10-hydroxycamptothecin, tert-butyl-4-aminopiperazine-1-carboxylate, methanol, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(4-Boc-piperazine-1-amine)methyl-10-hydroxycamptothecin (yield 87.8%). LC / MS(M+H): 578.26 (calculated value: 577.25).

[0144] Example 39: 9-((2-(2-Boc-aminoethoxy)ethyl)amine)methyl-10-hydroxycamptothecin (39) [ka] Formaldehyde, tert-butyl(2-(2-aminoethoxy)ethyl)carbamate, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-((2-(2-Boc-aminoethoxy)ethyl)amine)methyl-10-hydroxycamptothecin (yield 81.8%). LC / MS(M+H): 581.23 (calculated value: 580.25).

[0145] Example 40: 3-(N-Trifluoroethyl)-1,3-oxazine[5,6],[9,10]camptothecin (40)

Chem.

[0146] Example 41: 9-(Trifluoroethylamine)methyl-10-hydroxycamptothecin (41)

Chem.

[0147] Example 42: 9-(trans-4-Methoxycyclohexylamine)methyl-10-hydroxycamptothecin (42)

Chem.

[0148] Example 43: 9-(Tetrahydropyran-4-methylamine)methyl-10-hydroxycamptothecin (43)

Chem.

[0149] Example 44: 9-(N-methylhydroxyamine)methyl-10-hydroxycamptothecin (44) [ka] 9-Formyl-10-hydroxycamptothecin, N-methylhydroxyamine, methanol, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(N-methylhydroxyamine)methyl-10-hydroxycamptothecin (yield 44.1%). 1H NMR(500MHz,DMSO-d6)δ10.88(s,1H),8.98(s,1H),7.91(d,J=9.2Hz,1H),7.60(d,J=9.2Hz,1H),7.18(s,1H),6.46(s,1H),5.38(s,2 H),5.13(s,2H),4.71(s,1H),2.51(s,3H),1.83(hept,J=6.7,6.1Hz,2H),0.85(t,J=7.3Hz,3H);LC / MS(M+H):424.08(calculated value:423.14).

[0150] Example 45: 9-(2'-N-Boc-2'-N-methylhydrazine)methyl-10-hydroxycamptothecin (45) [ka] 9-Formyl-10-hydroxycamptothecin, tert-butyl-1-methylhydrazine-1-carboxylate, methanol, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(2'-N-Boc-2'-N-methylhydrazine)methyl-10-hydroxycamptothecin (yield 74.1%). 1 H NMR(600MHz,DMSO-d6)δ9.36(s,1H),8.71(s,1H),8.10(d,J=10.1Hz,2H),8.04(s,1H),7.55(d,J=9.2Hz,1H),7.28(s,1H),6.51(s ,1H),5.42(s,2H),5.27(s,2H),4.24(t,J=5.4Hz,1H),3.47(s,3H),1.87(dq,J=14.2,7.0Hz,2H),1.56(s,9H),0.91-0.87(m,3H). LC / MS (M+H): 523.15 (calculated value: 522.21).

[0151] Example 46: 9-(1-(4-hydroxyacetylpiperazine))methyl-10-hydroxycamptothecin (46) [ka] Compound 2 (15 mg, 3.2 mmol) was dissolved in THF (5 mL), hydroxyacetic acid (5.0 mg, 6.4 mmol) was added, and benzotriazole-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP, 2.9 mg, 6.4 mmol) was added while stirring. The mixture was stirred overnight at room temperature, the solvent was removed under reduced pressure, and the mixture was purified by silica gel column chromatography to obtain 9-(1-(4-hydroxyacetylpiperazine))methyl-10-hydroxycamptothecin (11.5 mg, yield 69.3%). 1 H NMR(500MHz,DMSO-d6)δ8.86(s,1H),8.09(d,J=9.2Hz,1H),7.58(d,J=9.2Hz,1H),7.27(s,1H),6.52(s,3H),5.42(s,2H),5.26(s,2H),4.43(s,2H) ),4.11(s,2H),3.56(d,J=43.9Hz,4H),3.15-2.89(m,4H),1.87(dt,J=16 .5,7.2Hz,2H),0.89(t,J=7.3Hz,3H);LC / MS(M+H):520.99(calculated value:520.20).

[0152] Example 47: 9-(N,O-dimethylhydroxyamine)methyl-10-hydroxycamptothecin (47) [ka] 9-Formyl-10-hydroxycamptothecin, N,O-dimethylhydroxyamine hydrochloride, methanol, and sodium cyanoborohydride were synthesized according to Method B to obtain compound 9-(N,O-dimethylhydroxyamine)methyl-10-hydroxycamptothecin (yield 65.3%). 11H NMR (600 MHz, DMSO-d6) δ 10.44 (s, 1H), 10.33 (s, 1H), 8.82 (s, 1H), 8.66 (s, 1H), 8.01 (dd, J = 15.4, 9.2 Hz, 1H), 7.52 (t, J = 8.5 Hz, 1H), 7.26 (d, J = 2.2 Hz, 1H), 6.49 (s, 1H), 5.42 (s, 2H), 5.27 (d, J = 4.5 Hz, 2H), 4.87 (s, 1H), 4.28 (s, 1H), 3.33 (d, J = 3.4 Hz, 3H), 2.55 (s, 1H), 1.86 (dh, J = 21.3, 7.2 Hz, 2H), 1.24 (s, 1H), 0.88 (t, J = 7.3 Hz, 3H). LC / MS (M+H): 438.11 (calculated value: 437.16).

[0153] Example 48: 9-((1-Methoxypropan-2-yl)amino)methyl-10-hydroxycamptothecin (48)

Chemical Structure

[0154] Example 49: 9-((1-Hydroxypropan-2-yl)amino)methyl-10-hydroxycamptothecin (49) [ka] Paraformaldehyde, 1-hydroxypropan-2-amine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-((1-hydroxypropan-2-yl)amino)methyl-10-hydroxycamptothecin (yield 84.7%). 1 H NMR(600MHz,DMSO-d6)δ11.42(s,1H),8.83(s,1H),8.71(s,1H),8.43(s,1H),8.15( d,J=9.2Hz,1H),7.63(d,J=9.1Hz,1H),7.29(s,1H),6.54(s,1H),5.50(s,1H),5.43 (s,2H),5.27(s,2H),4.61(s,2H),3.75(dd,J=11.7,4.5Hz,1H),3.63(dd,J=12.0,5 .8Hz,1H),1.87(m,J=21.4,7.2Hz,2H),1.34(d,J=6.6Hz,3H),0.89(t,J=7.3Hz,3H). LC / MS (M+H): 462.13 (calculated value: 461.48).

[0155] Example 50: 9-(propylamino)methyl-10-hydroxycamptothecin (50) [ka] Paraformaldehyde, n-propylamine, dioxane, and 10-hydroxycamptothecin were synthesized according to Method A to obtain compound 9-(propylamino)methyl-10-hydroxycamptothecin (yield 82.6%). 11H NMR (500 MHz, DMSO-d6) δ 11.29 (s, 1H), 8.87 (s, 1H), 8.63 (s, 2H), 8.17 (dd, J = 9.2, 1.7 Hz, 1H), 7.63 (dd, J = 9.2, 2.1 Hz, 1H), 7.29 (s, 1H), 6.52 (s, 1H), 5.43 (s, 2H), 5.29 (d, J = 2.8 Hz, 2H), 4.58 (d, J = 5.8 Hz, 2H), 3.03 (s, 2H), 1.93 - 1.80 (m, 2H), 1.71 (q, J = 7.7 Hz, 2H), 0.94 (t, J = 7.4 Hz, 3H), 0.89 (t, J = 7.3 Hz, 3H). LC / MS (M + H): 436.45 (calculated: 435.48).

[0156] Example 51: 9-((2-(2-Aminoethoxy)ethyl)amino)methyl-10-hydroxycamptothecin (51)

Chemical Structure

[0157] Test Example 1: Inhibitory activity of compounds against cancer cell proliferation Human esophageal cancer cells OE33, human breast cancer cells SK-BR-3, and human gastric cancer cells NCI-N87 were cultured in RPMI1640 (Cellmax) containing 10% fetal bovine serum (Cellmax). 1 × 10⁶ tumor cells in the culture medium were cultured in the exponential growth phase. 5 The test compound was diluted to cells / mL, and 100 μL was added per well to a 96-well cell culture plate. The plates were then returned to a 37°C, 5% CO2 incubator and incubated overnight. The following day, the test compound and control compound were diluted in culture medium to 10,000 nM, 2,000 nM, 400 nM, 80 nM, 16 nM, 3.2 nM, 0.64 nM, and 0.13 nM, respectively. 2 μL of the diluted compound was added per well to a 96-well cell culture plate, with three replication wells prepared for each concentration. 2 μL of the diluted solution was added per well to the negative control group and blank control group, which had not received any compound treatment. After addition, the plates were returned to a 37°C, 5% CO2 incubator and incubated for another 72 hours. After incubation, the cell culture plates were removed, the culture medium was removed by pipette, and 100 μL of medium containing 10% CCK-8 was added to each well. The plates were incubated at 37°C for 3 hours. After incubation was complete, the culture plate was removed, placed in an ELISA plate in the dark, and the absorbance was measured using 630 nm as the reference wavelength and 450 nm as the measurement wavelength. 50 The values ​​were calculated from absorbance values ​​using the 4-parameter regression method in GraphPad (Tables 1-3). SN38 (7-ethyl-10-hydroxycamptothecin) and 10CPT (10-hydroxycamptothecin) were used as control compounds. IC 50 Regarding the value, "++++" is IC 50 This indicates that it is <50nM, and "+++" is IC 50 This indicates that the current is between 10 and 100 nM, and "++" indicates IC 50 This indicates that the current is between 100 and 500 nM, and the "+" signifies IC 50 This indicates that the mass is >500 nM. [Table 2] [Table 3] [Table 4]

[0158] Results: The compounds of the examples provided by the present invention have a good inhibitory effect on the proliferation of cancer cells such as esophageal cancer cells OE33, breast cancer cells SK-BR-3, and gastric cancer cells NCI-N87. Several compounds, such as compound 10, compound 17, and compound 19, showed IC12 inhibition of the proliferation of these three types of cancer cells. 50 All values ​​are less than 100 nM, indicating broad anticancer activity.

Claims

1. The following compounds or pharmaceutically acceptable salts or stereoisomers thereof. 【Chemistry 1】

2. An antibody-drug conjugate comprising a low molecular weight drug and a linker antibody, characterized in that the low molecular weight drug is the compound described in Claim 1 or a pharmaceutically acceptable salt or stereoisomer thereof.

3. A pharmaceutical composition characterized by comprising the compound described in claim 1 or a pharmaceutically acceptable salt or stereoisomer thereof.

4. Use of the compound described in claim 1 or a pharmaceutically acceptable salt or stereoisomer thereof in the preparation of a drug for treating cancer.

5. Use of the antibody-drug conjugate according to claim 2 in the preparation of a drug for treating cancer.

6. Use of the pharmaceutical composition according to claim 3 in the preparation of a drug for treating cancer.

7. The use according to any one of claims 4 to 6, characterized in that the cancer includes gastric cancer, esophageal cancer, cardia cancer, breast cancer, ovarian cancer, colon cancer, rectal cancer, primary liver cancer, acute and chronic granulocytic leukemia, choriocarcinoma, lung cancer, and bladder cancer.

8. The use according to any one of claims 4 to 6, characterized in that the cancer is esophageal cancer, breast cancer, or gastric cancer.

9. The pharmaceutical composition according to claim 3 for treating cancer.

10. The pharmaceutical composition according to claim 9, characterized in that the cancer includes gastric cancer, esophageal cancer, cardia cancer, breast cancer, ovarian cancer, colon cancer, rectal cancer, primary liver cancer, acute and chronic granulocytic leukemia, choriocarcinoma, lung cancer, and bladder cancer.

11. The pharmaceutical composition according to claim 9, characterized in that the cancer is esophageal cancer, breast cancer, or gastric cancer.

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