Heterocycloalkenyl-substituted anthracycline derivative and use thereof

Abstract

Provided in the present application are a heterocycloalkenyl-substituted anthracycline derivative and the use thereof. Specifically, provided in the present application are a heterocycloalkenyl-substituted anthracycline derivative as represented by general formula (I) and the use thereof in the preparation of a drug for treating and / or preventing tumors.

Description

Heterocycloalkenyl-substituted anthracycline derivatives and uses thereof

[0001] This application claims priority to Chinese Patent Application No. 202411338856.X, filed on September 24, 2024, to Chinese Patent Application No. 202510097301.9, filed on January 21, 2025, and to Chinese Patent Application No. 202511316267.6, filed on September 15, 2025. This application incorporates the entirety of the above-mentioned Chinese patent applications. TECHNICAL FIELD

[0002] The present application belongs to the field of medicine, in particular to a heterocycloalkenyl-substituted anthracycline derivative and uses thereof. BACKGROUND

[0003] Antibody-drug conjugate (ADC) is a kind of targeted biological agent that couples target-specific monoclonal antibody (Ab) with high-killing cytotoxic drug (payload) through linker with specific cleavage properties. ADC uses monoclonal antibody as carrier to efficiently enrich small molecule cytotoxic drugs in target tumor cells, and then exerts anti-tumor effect. ADC combines the advantages of antibody with high selectivity and cytotoxic drug with high activity, retains the tumor-killing characteristics of small molecule cytotoxic drugs, selectively reduces the off-target toxic side effects of small molecule cytotoxic drugs, and effectively improves the benefit-risk ratio of anti-tumor therapy. A representative ADC drug is DS-8201 (trade name: ) developed by Dainippon Sumitomo Pharma, which was approved by FDA in 2019 for the treatment of HER2-positive breast cancer, and subsequently approved for the treatment of various solid tumors. DS-8201 uses trastuzumab as the HER2-targeting antibody component, and the payload molecule Dxd is a topoisomerase I inhibitor, which is the main cell-killing active ingredient.

[0004] Anthracycline derivatives as topoisomerase II inhibitors are a major class of chemotherapeutic drugs used in clinic. The anthracycline drugs that have been marketed include doxorubicin, daunorubicin, epirubicin, idarubicin, aclarubicin, valrubicin, pirarubicin and amrubicin. Doxorubicin, as a classic broad-spectrum antitumor drug, is often used in combination with other chemotherapeutic drugs to treat various hematological tumors and solid tumors. Doxorubicin is one of the earliest chemotherapeutic drugs used as a toxin carrier in the field of ADC research. ADC molecules with doxorubicin as a carrier include BR96-DOX and hLL1-DOX, both of which have been evaluated in clinic, but the relevant clinical trials were terminated due to insufficient efficacy, mainly because the antitumor activity of doxorubicin itself is too weak. PNU-159682 is the active metabolite of nemorubicin, and its in vitro activity is more than 3000 times stronger than that of doxorubicin. NBE Therapeutics developed an ADC technology platform based on PNU-159682, and its related product NBE-002 was evaluated in clinic, but the relevant trial was terminated in clinical phase I, which may be due to the strong clinical toxicity of PNU-159682 and its derivatives. Clinically commonly used chemotherapeutic drugs mainly include alkylating agents, Topo I inhibitors, microtubule protein inhibitors and Topo II inhibitors in terms of mechanism. The first three types of small molecule compounds have been developed into ADC carriers for clinical use, and only Topo II inhibitor compounds have not been successfully developed into ADC carriers for clinical use. In view of the broad-spectrum antitumor properties of anthracyclines and the previous exploration of anthracycline-based ADC drug development in the field, it is necessary to develop new anthracycline compounds as ADC carriers to balance the efficacy and toxicity of anthracycline ADCs, and further tap the clinical (indication) development potential of anthracycline ADCs.

[0005] The present application aims to develop a class of anthracycline derivatives with moderate activity to meet the development needs of new generation anthracycline ADC carrier molecules. SUMMARY

[0006] The first aspect of the present application provides a compound represented by formula (I), a pharmaceutically acceptable salt thereof or a stereoisomer thereof:

[0007] wherein,

[0008] R1is selected from H, hydroxyl and C 1-6 alkoxy;

[0009] R2is selected from O and NH;

[0010] R3is selected from C 1-6 alkyl, hydroxyc 1-6 alkyl, -C(O)C 1-6 alkyl and -C(O)NR n1 Rn2 ;

[0011] R n1 and R n2 are each independently selected from the group consisting of H, optionally substituted C n2-1 alkyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 4- to 10-membered bridged cycloalkyl, optionally substituted 3- to 8-membered heterocyclyl, optionally substituted 5- to 10-membered heteroaryl, and optionally substituted 6- to 10-membered aryl; 1-6 n2-2 n2-3 n2-4 n2-5 n2-6

[0012] each R n2-1 , R n2-2 , R n2-3 , R n2-4 , R n2-5 , and R n2-6 are each independently selected from the group consisting of hydroxy, C 1-6 alkyl, C 1-6 alkoxy, halogen, amino, hydroxyC 1-6 alkyl, -NH-C 1-6 alkyl, C 1-6 alkyl-NH-C 1-6 alkyl, nitro, cyano, halogenated C 1-6 alkyl, alkinyl, alkenyl, optionally substituted 3- to 8-membered heterocyclyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 6- to 10-membered aryl, and optionally substituted 5- to 10-membered heteroaryl; n2-1-1 n2-1-2 n2-1-3 n2-1-4 ;

[0013] each R n2-1-1 , R n2-1-2 , R n2-1-3 , R n2-1-4 is each independently selected from the group consisting of halogen, C 1-6 alkyl, amino, -NH-C 1-6 alkyl, C 1-6 alkyl-NH2, nitro, cyano, and halogenated C 1-6 alkyl;

[0014] or

[0015] R n1 and R n2 together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl, said 3- to 8-membered heterocyclyl being optionally substituted with C​​​​​​​​​1-6 Alkyl, C 1-6 Alkoxy, amino, hydroxyl, -NH-C 1-6 Alkyl, C 1-6 Alkyl-NH-C 1-6 Alkyl, hydroxy C 1-6 Alkyl, nitro, cyano, trifluoromethyl, difluoromethyl, monofluoromethyl, trichloromethyl, alkynyl, alkenyl substitution;

[0016] R4 and R5 together with the nitrogen atoms they are attached to form 3 to 8-membered heterocyclic alkenyl groups.

[0017] In some embodiments of this application, R1 is selected from -OCH3.

[0018] In some embodiments of this application, R2 is selected from O.

[0019] In some embodiments of this application, R3 is selected from hydroxyC 1-6 Alkyl and -C(O)NR n1 R n2 .

[0020] In some embodiments of this application, R n1 Selected from H and C 1-6 Alkyl group; preferably H.

[0021] In some embodiments of this application, R n2 Selected from one or more R n2-1 Replacement C 1-6 Alkyl, optionally with one or more R n2-2 Substituted 3- to 8-membered cycloalkyl groups, optionally with one or more R n2-3 Replacement of 4- to 10-membered bridged ring groups, 3- to 8-membered heterocyclic groups, and optionally replaced by one or more R n2-6 The substituted 6-10 aryl group; preferably selected from those substituted by one or more R n2-1 Replacement C 1-6 Alkyl groups and optionally one or more R n2-6 Replaced 6-10 aryl groups.

[0022] In some embodiments of this application, each R n2-1 Each is independently selected from hydroxyl, amino, -NH-C 1-6 Alkyl, 3 to 8-membered heterocyclic group, with one or more R n2-1-1 The substituted 3 to 8-membered heterocyclic group, optionally replaced by one or more R n2-1-2 Substituted 3- to 8-membered cycloalkyl groups and optionally with one or more R n2-1-3 Substituted 6-10 aryl groups; preferably selected from hydroxyl, -NH-C 1-6 Alkyl, with one or more R n2-1-1substituted 3- to 8-membered heterocyclyl and optionally substituted by one or more R n2-1-3 substituted 6- to 10-membered aryl.

[0023] In certain embodiments of the application, each R n2-1-1 each independently selected from halogen and C 1-6 alkyl.

[0024] In certain embodiments of the application, each R n2-1-2 each independently amino.

[0025] In certain embodiments of the application, each R n2-1-3 each independently selected from halogen, C 1-6 alkyl, amino, -NH-C 1-6 alkyl, C 1-6 alkyl-NH2, nitro, cyano and halogenated C 1-6 alkyl; preferably selected from halogen, C 1-6 alkyl, amino, -NH-C 1-6 alkyl and C 1-6 alkyl-NH2.

[0026] In certain embodiments of the application, each R n2-1-4 each independently amino.

[0027] In certain embodiments of the application, each R n2-2 each independently selected from amino, hydroxy C 1-6 alkyl, -NH-C 1-6 alkyl; preferably amino.

[0028] In certain embodiments of the application, each R n2-3 and R n2-5 each independently amino.

[0029] In certain embodiments of the application, each R n2-4 each independently selected from amino, C 1-6 alkyl-NH2and C 1-6 alkyl.

[0030] In certain embodiments of the application, each R n2-6 each independently selected from halogen, C 1-6 alkyl, amino, C 1-6 alkyl-NH-C 1-6 alkyl, -NH-C 1-6 alkyl, C 1-6 alkyl-NH2, nitro, cyano and halogenated C 1-6 alkyl; preferably selected from halogen, C 1-6 alkyl, amino, C 1-6alkyl-NH-C 1-6 alkyl and -NH-C 1-6 alkyl.

[0031] In certain embodiments of the application, R n1 and R n2 together with the nitrogen atom to which they are attached form a 3- to 8- membered heterocyclyl group, which is optionally substituted with -NH-C 1-6 alkyl.

[0032] In certain embodiments of the application, in the 3- to 8- membered heterocyclyl group, the heteroatom(s) is one or more of N, O and S, the number of heteroatoms being 1 or 2; preferably, the heteroatom is N.

[0033] In certain embodiments of the application, in the 5- to 10- membered heteroaryl group, the heteroatom(s) is one or more of N, O and S, the number of heteroatoms being 1 or 2; preferably, the heteroatom is N and / or O.

[0034] In certain embodiments of the application, R3is selected from

[0035] In certain embodiments of the application, R3is selected from

[0036] In certain embodiments of the application, R4and R5together with the nitrogen atom to which they are attached form a 5- to 6- membered heterocyclenyl group.

[0037] In certain embodiments of the application, the fragment -NR4R5is selected from the following structures:

[0038] preferably

[0039] In certain embodiments of the application, the compound of formula (I) is a compound of formula (IA):

[0040] wherein R1, R2, R3, R4, R5are as defined in any one of the above.

[0041] In certain embodiments of the application, the compound of formula (I), the pharmaceutically acceptable salt thereof, the stereoisomer thereof, is selected from the compounds of formula (I-1), formula (I-2), formula (I-3), the pharmaceutically acceptable salt thereof, the stereoisomer thereof:

[0042] In the formula, R3 is as defined in any of the preceding technical solutions.

[0043] In another aspect, the present application provides a new anthracycline compound, a pharmaceutically acceptable salt thereof, an isomer thereof, which is selected from the group consisting of:

[0044] In another aspect, the present application provides the above-mentioned compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, for use in the treatment and / or prevention of a tumor.

[0045] In another aspect, the present application provides an antibody drug conjugate comprising a small molecule drug, a linker and an antibody, wherein the small molecule drug is selected from the above-mentioned compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof.

[0046] In another aspect, the present application provides a pharmaceutical composition comprising the above-mentioned compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, and one or more pharmaceutically acceptable carriers.

[0047] In another aspect, the present application provides the use of the above-mentioned compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or the above-mentioned pharmaceutical composition in the manufacture of a medicament for the treatment and / or prevention of a tumor.

[0048] In certain embodiments of the present application, the tumor is breast cancer, lung cancer or colorectal cancer.

[0049] In another aspect, the present application provides a method for treating and / or preventing a tumor, the method comprising administering to a subject in need thereof an effective amount of the above-mentioned compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or the above-mentioned pharmaceutical composition.

[0050] In certain embodiments of the present application, the tumor is breast cancer, lung cancer or colorectal cancer.

[0051] In certain embodiments of the present application, the content of the compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof in the pharmaceutical composition is 1% to 95%.

[0052] In certain embodiments of the present application, the pharmaceutically acceptable carrier in the pharmaceutical composition comprises one or more of a filler, a disintegrant, a binder, a glidant, a lubricant.

[0053] Terminology

[0054] The following terms and phrases, as used in the present application, are intended to have the following meanings unless otherwise indicated. A particular term or phrase should not be construed as undefined or unclear without a specific definition, but should be understood according to the ordinary meaning.

[0055] The term "pharmaceutically acceptable" means, within the scope of sound medical judgment, those compounds, materials, compositions, and / or dosage forms which are suitable for use with human and animal tissues without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0056] The term "pharmaceutically acceptable salt" means a derivative of the compounds of the present application produced by reaction with a relatively nontoxic acid or base. These salts can be prepared in situ during the synthesis, isolation, and purification of the compound, or separately by reacting the purified compound with a suitable acid or base. When the compound contains relatively acidic functional groups (e.g., -COOH, -OH, -SO3H, etc.), base addition salts can be formed by reaction with appropriate inorganic or organic cations (bases), including alkali or alkaline earth metal salts, ammonium salts, salts with amines or derivatives thereof, salts with amino acids, etc. When the compound contains relatively basic functional groups (e.g., -NH2, etc.), acid addition salts can be formed by reaction with appropriate inorganic or organic acids (e.g., carboxylic acids, etc.).

[0057] The term "pharmaceutically acceptable carrier" means a medium generally accepted in the art for delivery of biologically active agents to animals, particularly mammals, and includes to, e.g., adjuvants, excipients or vehicles, such as diluents, preservatives, fillers, flow conditioners, disintegrants, wetting agents, emulsifiers, suspending agents, sweetening agents, flavoring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents, and dispersing agents, depending upon the nature of the dosage form and the means of administration. Pharmaceutically acceptable carriers are formulated in accordance with routine procedures, as described in the art, based on the nature of the active agent being delivered. They include, but are not limited to: the type and nature of the active agent being formulated; the subject to whom the composition containing the agent is to be administered; the intended route of administration of the composition; and the target therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous media, as well as a variety of solid and semi-solid dosage forms. Such carriers include a wide variety of different ingredients and additives, which are included in the formulation for a variety of reasons (e.g., to stabilize the active agent, to bind the agents, etc.), and such additional ingredients are well known to those of ordinary skill in the art.

[0058] The term "effective prophylactic or therapeutic treatment amount" means a sufficient amount of a compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof to treat a disorder at a reasonable benefit / risk ratio applicable to any medical treatment and / or prophylaxis. It will be understood, however, that the total daily usage of the compounds of the formula I, or their pharmaceutically acceptable salts, and compositions of the present application will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex, and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.

[0059] The term "isomers" as used herein includes geometric isomers and stereoisomers, such as atropisomers, cis-trans isomers, enantiomers, diastereomers, tautomers, as well as racemic mixtures and other mixtures of the foregoing, all of which are intended to be within the scope of the present application. The term "enantiomers" refers to stereoisomers that are mirror images of one another. The term "tautomers" refers to functional group isomers that differ in the bonding of one or more atoms due to the migration of a proton or a bond. For example, a ketone and its enol form are keto-enol tautomers. The term "diastereomers" refers to stereoisomers that have two or more chiral centers and are not mirror images of one another. The term "cis-trans isomers" refers to different spatial orientations of a double bond or ring atom single bond that cannot freely rotate. The term "atropisomers" refers to stereoisomers that are separated due to hindered rotation or very slow rotation of a single bond. Stereoisomers of the compounds of the present application can be prepared using chiral synthesis or chiral reagents or other conventional techniques. For example, one enantiomer of a compound of the present application can be prepared by asymmetric catalytic techniques or chiral auxiliary derivatization techniques. Alternatively, a single stereoisomer can be obtained from a mixture by chiral resolution techniques. Alternatively, a single stereoisomer can be prepared directly using a chiral starting material. Separation of optically pure compounds is typically accomplished using preparative chromatography on chiral columns to achieve separation of the chiral compounds.

[0060] The absolute stereochemistry is specified using the configuration of the compound of the present application with the skilled person being aware of the possibility of interconversion of the compounds of the present application. The absolute stereochemistry of the compounds of the present application can be established by conventional techniques. For example, single crystal X-ray diffraction, by the chiral structure of the starting material and the reaction mechanism of asymmetric synthesis. Alternatively, after resolution, by comparison with the product of absolute configuration. Compounds of the present application marked as "absolute configuration unknown / undetermined" are typically resolved from racemic compounds by chiral preparative SFC to a single isomer, which is then characterized and tested.

[0061] It is known to those skilled in the art that a cyclic compound has aromaticity when it has a planar delocalized system and the number of π-electrons is 4n+2. The representation of the aromatic structure in the compound can be either by dotted lines to indicate electron delocalization or by single and double bonds in an alternating manner, such as in the benzene ring, which can be represented as or as

[0062] The term "optionally substituted" as used herein means that one or more hydrogen atoms of the group to which this term applies can be "replaced" by one or more substituents or "not replaced".

[0063] When a bond in a substituent group is interrupted by , it means that the bond is the connecting bond of the substituent, for example , which means that the pyrimidine ring is connected to the given group or the given structural formula via the C atom. The occurrence of a dash "-" in a substituent group indicates the point of attachment for the substituent, for example -SCH3, which is connected to the given group or the given structural formula via the S atom. and , which means the absolute configuration of a stereogenic center, i.e. the R or S configuration. or , which means the cis or trans configuration. A double bond represented by a double bond or a single bond represents the cis configuration, while a single bond and a double bond represent the trans configuration.

[0064] When a bond of a substituent can crosslink to a ring, it means that this substituent can bond to any atom of the ring. For example, the structural element means that the substituent R can be substituted at any position on the benzene ring.

[0065] When a listed substituent is not specified as to which atom of the substituent is connected to the given group or the given structural formula, the substituent can be connected via any bondable atom thereof.

[0066] When any variable (for example R d ) occurs more than one time in a compound or a structural formula, its definition on each occurrence is independent of its definition at every other occurrence. For example, means that the cyclopentyl group is substituted with 3 R d groups, and each R d group is independently selected.

[0067] Unless otherwise specified, the term "halogen" means a fluorine, chlorine, bromine, or iodine atom.

[0068] Unless otherwise specified, the term "alkyl" means a branched or straight-chain saturated aliphatic hydrocarbon derived from a parent alkane by removal of a single hydrogen atom. For example, "C 1-10 "alkyl" means a branched or straight-chain saturated aliphatic hydrocarbon group having the indicated number of carbon atoms. For example, "C 10 "alkyl" means a branched or straight-chain saturated aliphatic hydrocarbon group having the indicated number of carbon atoms. For example, "C 1-6 "alkyl" means a branched or straight-chain saturated aliphatic hydrocarbon group having the indicated number of carbon atoms. For example, "C 1- "alkyl" means a branched or straight-chain saturated aliphatic hydrocarbon group having the indicated number of carbon atoms. For example, "C 1-3 "alkyl" means a branched or straight-chain saturated aliphatic hydrocarbon group having the indicated number of carbon atoms. For example, "C

[0069] Unless otherwise specified, the term "haloalkyl" means an alkyl group in which one or more hydrogen atoms are replaced by a halogen atom. Preferably, the "haloalkyl" groups described herein are "haloC 1-6 "alkyl", more preferably "haloC 1-4 "alkyl". Examples of haloalkyl groups include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, and the like. Alkyl groups are as previously defined.

[0070] Unless otherwise specified, the term "hydroxyalkyl" means a group derived from an alkyl group in which one or more hydrogen atoms have been replaced by a hydroxyl group. The "hydroxyalkyl" groups described herein include "hydroxyC 1-6 "alkyl", "hydroxyC 1-4 "alkyl"; specific examples include, but are not limited to, -CH2OH, -CH2CH2OH, -CH(OH)CH3, -CH2CH2CH2OH, and the like.

[0071] Unless otherwise specified, the term "alkoxy" means an alkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom, i.e., "alkyl-O-". The "alkoxy" groups described herein include "C 1-6 "alkyl" (having the structure C 1-6 "alkyl-O-"), "C 1-4 "alkyl"; specific examples include, but are not limited to, methoxy, ethoxy, propyloxy, 1-methylethoxy, butoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropyloxy, 1,2-dimethylpropyloxy, and the like; preferably, the "alkoxy" groups described herein are "C 1-4 "alkyl", more preferably "C 1-3 "alkyl".

[0072] Unless otherwise specified, the term "haloalkoxy" means an alkoxy group in which one or more hydrogens have been replaced by a halogen. Preferably, the "haloalkoxy" groups described herein are "haloC 1-6 "alkyl", "haloC 1-4"Alkoxy". Specific examples of "alkoxy" groups as used herein include methoxy, ethoxy, propyloxy, isopropyloxy, butyloxy, isobutyloxy, pentyloxy, isopentyloxy, hexyloxy, cyclohexyloxy, heptyloxy, cycloheptyloxy, octyloxy, cyclooctyloxy, nonyloxy, decyloxy, cyclodecyloxy, and the like. Alkoxy groups are defined as O-alkyl, wherein "alkyl" is as previously defined.

[0073] Unless otherwise specified, the term "ring" refers to saturated, partially saturated, or unsaturated monocyclic and polycyclic rings, "polycyclic" including spiro, fused, or bridged rings. A group derived from a ring by removal of a hydrogen atom is referred to as a "ring group", which includes monovalent rings, divalent rings (often referred to as "ring alkenes"), trivalent rings, tetravalent rings, and the like, depending on the number of substituents attached to the ring. The present application does not specifically distinguish the valence of the "ring group". Representative "ring groups" include substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl, or heteroaryl.

[0074] The term "hetero" refers to substituted or unsubstituted heteroatoms and oxidized forms of heteroatoms (also referred to as heteroatom groups), which are generally selected from N, O, S, P, and oxidized forms generally include NO, SO, S(O)2, P(O), and the nitrogen atom can be substituted, i.e., NR (R is H or other substituents as defined herein); the number of atoms in a ring is generally defined as the number of members of the ring, e.g., "3-6 membered heterocycloalkyl" refers to a ring of 3-6 atoms arranged in a ring, each ring optionally containing 1-3 heteroatoms and / or heteroatom groups, i.e., N, O, S, NO, SO, S(O)2, P(O), or NR, each ring optionally substituted with R groups, R being a group as defined herein.

[0075] Unless otherwise specified, the term "cycloalkyl" refers to saturated cyclic alkyl groups derived from cycloalkanes, including monocyclic or polycyclic saturated hydrocarbon groups; the polycyclic saturated hydrocarbon groups refer to polycyclic groups formed by two or more cyclic alkyl structures connected by spiro, bridge, fusion, and the like. The carbon atoms in the cycloalkyl group can be further oxidized, i.e., C(O). Unless otherwise specified, "cycloalkyl" as used herein can be understood as monocyclic cycloalkyl groups, and when polycyclic, it will be specifically indicated as spiro, fused, or bridged groups. The cycloalkyl groups include "3 to 10 membered cycloalkyl", "5 to 10 membered cycloalkyl", "3 to 8 membered cycloalkyl", "4 to 8 membered cycloalkyl", "3 to 6 membered cycloalkyl", "3 to 5 membered cycloalkyl". Preferably, the cycloalkyl groups are monocyclic, saturated structures, such as C 3-8 Cycloalkyl groups; specific examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl.

[0076] Unless otherwise specified, "cycloalkenyl" means one or more of the ring-forming bonds in "cycloalkyl" are double bonds and the cycloalkenyl group is not aromatic. The carbon atoms in the cycloalkenyl group can be further oxidized, i.e., form C(O). The cycloalkenyl groups include "3 to 10 membered cycloalkenyl", "5 to 10 membered cycloalkenyl", "3 to 8 membered cycloalkenyl", "3 to 6 membered cycloalkenyl", "3 to 5 membered cycloalkenyl", "5 to 6 membered cycloalkenyl". Specific examples include, but are not limited to

[0077] Unless otherwise specified, the term "heterocyclyl" means a saturated ring group derived from replacing one or more ring carbon atoms in cycloalkyl with a heteroatom and / or heteroatom group. The heteroatom and / or heteroatom group is typically selected from N, O, S, NO, SO, S(O)2, P(O), and NR, wherein the carbon atoms in the heterocycle are optionally oxidized, i.e., form -C(O); preferably, the heteroatoms are independently selected from 1-3 N and / or O. The heterocyclyl groups include "3 to 10 membered heterocyclyl", "5 to 10 membered heterocyclyl", "3 to 8 membered heterocyclyl", "3 to 6 membered heterocyclyl", "3 to 5 membered heterocyclyl", "4 to 8 membered heterocyclyl", "4 to 6 membered heterocyclyl", "5 to 6 membered heterocyclyl", "4 membered heterocyclyl", "5 membered heterocyclyl". Specific examples include, but are not limited to, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, and the like.

[0078] Unless otherwise specified, the term "heterocycloalkenyl" means one or more of the ring-forming bonds in "heterocyclyl" are double bonds and the heterocycle is not aromatic. Preferably, the heteroatoms are independently selected from 1-3 N and / or O. The heterocycloalkenyl groups include "3-8 membered heterocycloalkenyl", "3-6 membered heterocycloalkenyl", "3-5 membered heterocycloalkenyl", "5-6 membered heterocycloalkenyl". Specific examples include, but are not limited to: and the like.

[0079] Unless otherwise specified, the term "aryl" means an unsaturated, usually aromatic, hydrocarbon group which can be a single ring or multiple rings which are fused together. Preferably 5 to 10 membered aryl, more preferably 5 to 8 membered aryl, most preferably a single ring 5 to 6 membered aryl; examples of aryl groups include, but are not limited to, phenyl, naphthyl.

[0080] "Heteroaryl" as used herein refers to a monocyclic or polycyclic group having aromaticity, containing one or more heteroatoms in the ring, which heteroatoms are typically selected from N, O, S; preferably, the heteroatoms are independently selected from 1-3 N and / or O, and additionally, the N and S atoms can be optionally oxidized and the N atoms can be optionally quaternized. "Heteroaryl" includes "monocyclic heteroaryl" and "fused heteroaryl", the latter referring to a group having aromaticity as a whole, containing one or more heteroatoms, formed by two or more cyclic structures sharing two adjacent atoms with each other. Unless specifically indicated, "heteroaryl" as used herein is generally understood to be "monocyclic heteroaryl", for example, "5-6 membered heteroaryl" as used herein does not have the potential to form a fused heteroaryl group; when it is a "fused heteroaryl", it will be specifically indicated as "8-10 membered fused heteroaryl". Preferably, the heteroaryl group as used herein is a "nitrogen-containing heteroaryl", preferably a "5-6 membered nitrogen-containing heteroaryl", the "nitrogen-containing heteroaryl" contains at least one nitrogen atom, for example, contains only 1, 2 or 3 nitrogen atoms, or contains one nitrogen atom and one or two other heteroatoms (e.g., S and / or O atoms), or contains two nitrogen atoms and one or two other heteroatoms. Specific examples of the heteroaryl group include, but are not limited to, furanyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, and the like.

[0081] Unless otherwise specified, the term "bridged cyclic group" refers to a saturated or partially saturated cyclic structure formed by two or more carbocyclic rings sharing two non-adjacent carbon atoms with each other. Optionally, a carbon atom in the cyclic structure can be oxidized. "Bridged cyclic group" includes, for example, "4-10 membered bridged cyclic group", "4-5 membered bridged cyclic group", "5-11 membered bridged cyclic group", "6-11 membered bridged cyclic group", "5-10 membered bridged cyclic group", "5-7 membered bridged cyclic group", "7-10 membered bridged cyclic group", "6-9 membered bridged cyclic group", "7-8 membered bridged cyclic group", "9-10 membered bridged cyclic group", and the like. Specific examples include, but are not limited to:

[0082] Unless otherwise specified, the term "bridged heterocyclyl" refers to a saturated or partially saturated ring structure derived by replacing at least one carbon atom in a "bridged cycloalkyl" with a heteroatom / heteroatom group selected from the group consisting of N, O, S, P, NO, SO, S(O)2, P(O), and NR, R being H or any substituent that can be present, said "bridged cycloalkyl" being as defined above. "Bridged heterocyclyl" includes, for example, "4-10 membered bridged heterocyclyl", "5-11 membered bridged heterocyclyl", "6-11 membered bridged heterocyclyl", "5-10 membered bridged heterocyclyl", "7-10 membered bridged heterocyclyl", "6-9 membered bridged heterocyclyl", "7-8 membered bridged heterocyclyl", "9-10 membered bridged heterocyclyl", "6-10 membered bridged heterocycloalkenyl", "6-8 membered bridged heterocycloalkenyl", "7-8 membered nitrogen-containing bridged heterocycloalkenyl", and the like. Preferably, the heteroatoms are independently selected from 1-3 N and / or O. Preferably, the bridged heterocyclyl is a "nitrogen-containing bridged heterocyclyl", meaning that at least one ring atom is N, and optionally contains one or more additional heteroatoms; preferably, the "nitrogen-containing bridged heterocyclyl" contains 1 N atom and 0-2 atoms selected from N and / or O and / or S. Preferably, the "nitrogen-containing bridged heterocyclyl" contains 1 N atom and 0-1 atoms selected from O and / or S. Preferably, the bridged heterocyclyl is an "oxygen-containing bridged heterocyclyl", meaning that at least one ring atom is O, and optionally contains one or more additional heteroatoms; preferably, the "oxygen-containing bridged heterocyclyl" contains 1 O atom and 0-2 atoms selected from N and / or O. Specific examples include, but are not limited to: and the like.

[0083] When a bond of a substituent can cross-link to a ring, it means that the substituent can bond to any atom on the ring. For example, the structural unit means that the substituent R b can be substituted at any position on ring B, including NH, and R b is n in number; when the ring contains NH, it means that NH and other ring atoms can be substituted by R b , i.e. R b may or may not substitute H.

[0084] When any variable (e.g. R b ) appears more than once in a compound or a structure, each definition is independent. For example, the structural unit means that ring B is substituted by n R b , and each R b has an independent option.

[0085] When a listed substituent is not specified as to its attachment point to a given group or given formula, then the substituent can be attached by any of its available bonding sites. For example, pyrimidine as a substituent means that the pyrimidine ring is attached to the group being substituted through any one of the carbon atoms or the nitrogen atoms, and further for example, ring B in the present application is selected from When a listed substituent is not specified as to its attachment point to a given group or given formula, then the substituent can be attached by any of its available bonding sites. For example, pyrimidine as a substituent means that the pyrimidine ring is attached to the group being substituted through any one of the carbon atoms or the nitrogen atoms, and further for example, ring B in the present application is selected from

[0086] It is specifically contemplated that all combinations of substituents and / or variables in the present application are permissible unless such combinations result in unsound compounds.

[0087] It is specifically contemplated that all combinations of substituents and / or variables in the present application are permissible unless such combinations result in unsound compounds.

[0088] "Substituted" or "substitution" means that one or more hydrogen atoms, preferably 1 to 6, more preferably 1 to 3, of a group are each, independently of one another, replaced with the corresponding number of substituents. One skilled in the art can determine, without undue effort (either experimentally or theoretically), what substitutions are possible or impossible. For example, an amino or hydroxyl group with a free hydrogen can not be stable when bound to a carbon atom with an unsaturated (e.g., olefinic) bond.

[0089] "Pharmaceutical composition" means a mixture of one or more of the compounds described herein, or physiologically / pharmaceutically acceptable salts or prodrugs thereof, with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to a subject, and to facilitate absorption, resulting in the active ingredient exerting its biological activity.

[0090] The term "pharmaceutically acceptable salt" refers to derivatives of the compounds of the application formed with relatively nontoxic inorganic or organic acids or bases. These salts can be prepared during the final isolation and purification of the compounds or separately by reacting the purified compound with the appropriate acid or base, for example. When the compound contains a relatively acidic functionality, base addition salts can be prepared by contacting the neutral form of the compound with a solution of a physiologically / pharmaceutically acceptable base, including metal and ammonium hydroxides and organic amines. When the compound contains a relatively basic functionality, acid addition salts can be prepared by contacting the neutral form of the compound with a solution of a physiologically / pharmaceutically acceptable acid.

[0091] When the term "about" is applied to a parameter such as pH, concentration, temperature, etc., it indicates that the parameter can vary by ±10%, and sometimes more preferably within ±5%. As will be understood by those skilled in the art, numbers are often given only to the nearest integer for illustrative purposes, not as a limitation.

[0092] The preparation methods of some compounds in this application refer to the preparation methods of the aforementioned analogous compounds. Those skilled in the art should know that when using or referring to using the preparation methods cited thereby, the feeding ratio of reactants, reaction solvent, reaction temperature, etc. can be appropriately adjusted according to the different reactants.

[0093] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by the combination of the specific embodiments with other chemical synthetic methods well known to those skilled in the art, and equivalent replacement methods well known to those skilled in the art, preferred embodiments including but not limited to the examples of the present application.

[0094] Summary of experimental instruments:

[0095] The structure of the compounds of the present application is determined by nuclear magnetic resonance (NMR) or / and liquid chromatography-mass spectrometry (LC-MS), or ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). The NMR chemical shift (δ) is given in units of parts per million (ppm). The NMR determination is made using a Bruker Neo400M or Bruker Ascend 400 nuclear magnetic instrument, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD) and deuterated chloroform (CDCl3), heavy water (D2O) as the determination solvent, and tetramethylsilane (TMS) as the internal standard.

[0096] The determination of liquid chromatography-mass spectrometry (LC-MS) is made using an Agilent 1260-6125B single quadrupole mass spectrometer, with a Welch Biomate column (C18, 2.7 μm, 4.6 x 50 mm) or a waters H-Class SQD2, with a Welch Ultimate column (XB-C18, 1.8 μm, 2.1 x 50 mm) mass spectrometer (ion source is electrospray ionization).

[0097] The determination of ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) is made using a Waters UPLC H-class SQD mass spectrometer (ion source is electrospray ionization).

[0098] The determination of HPLC is made using a Waters e2695-2998 or Waters ARC and Agilent 1260 or Agilent Poroshell HPH high-performance liquid chromatograph.

[0099] Preparative HPLC used Waters 2555-2489 (10 pm, ODS250 cm x 5 cm) or GILSON Trilution LC, column was Welch XB-C18 column (5 pm, 21.2 x 150 mm).

[0100] Thin layer chromatography silica gel plate (TLC) used Yantai Jiangyou Silica Gel Development Co., Ltd. GF254 silica gel plate or Rushan Shangbang New Material Co., Ltd. GF254 silica gel plate, TLC used specifications were 0.15 mm to 0.20 mm, preparative 20 x 20 cm, column chromatography generally used 200-300 mesh silica gel as carrier. DETAILED DESCRIPTION

[0101] The starting materials in the embodiments of the present application are known and commercially available, or can be synthesized by using or according to the methods known in the art. Unless otherwise specified, all reactions in the present application are carried out under continuous magnetic stirring, the solvent is dry solvent, and the reaction temperature unit is Celsius or °C. The room temperature generally refers to 15-35 °C, preferably 20-30 °C, more preferably 20-25 °C. It will be apparent to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present application without departing from the spirit and scope of the present application.

[0102] Example 1

[0103] Synthesis of (8R,10S)-10-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H-pyrrol-1-yl)-5- hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-8-(2- hydroxyethyl)-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione (Compound DP-1)

[0104] Step one:

[0105] Synthesis of N'-((E)-1-((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydrotetracene-2-yl)ethenediamine (Compound 1-2)

[0106] Benzene sulfonyl hydrazide (375 mg, 2.18 mmol) and doxorubicin hydrochloride (500 mg, 0.86 mmol) were dissolved in anhydrous methanol (15 mL) at room temperature. The system was heated to 45 °C and stirred for 20 h. LC-MS was used to monitor the reaction. The reaction was complete. 100 mL methyl tert-butyl ether was added to the reaction system to precipitate the product. The suspension was transferred to a centrifuge tube and centrifuged (2000 rpm, 10 min). The supernatant was discarded and the precipitate was washed with methyl tert-butyl ether and centrifuged. The precipitate was air-dried to obtain 600 mg of the brown red target product.

[0107] Step two:

[0108] Synthesis of (8R,10S)-10-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,8,11-trihydroxy-8-(2-hydroxyethyl)-1-methoxy-7,8,9,10-tetrahydrobenzo[cd]indolizine-5,12-dione (Compound 1-3)

[0109] Compound 1-2 (600 mg, 0.85 mmol) was dissolved in MeOH (45 mL) at room temperature. NaCNBH3 (641 mg, 10.2 mmol) and p-toluenesulfonic acid pyridine (429 mg, 1.7 mmol) were added to the solution under ice bath. The reaction was refluxed for 30 min, then cooled to room temperature and poured into dichloromethane (200 mL). The reaction solution was washed with water and saturated brine successively, dried over anhydrous sodium sulfate and filtered under reduced pressure. The filtrate was concentrated under reduced pressure. The residue was separated and purified by C18 reverse phase column chromatography (acetonitrile-0.05% ammonium acetate aqueous solution, 5%-95%) to obtain 180 mg of the red target compound.

[0110] Step three:

[0111] Synthesis of Compound DP-1

[0112] Compound 1-3 (180 mg, 0.34 mmol) was dissolved in DMF (2.5 mL) at room temperature. 5-iodopentanal (1.8 g, 8.5 mmol) and DIPEA (180 μL, 1.03 mmol) were added to the reaction solution successively. The reaction was stirred for 10 min. LC-MS was used to monitor the reaction. The reaction was complete. The reaction solution was purified by HPLC preparation (acetonitrile-0.1% TFA aqueous solution, 15%-35%, 30 mL / min) to obtain 93 mg of the brown red title compound.

[0113] LC-MS: 582.3 [M+H] +

[0114] 1H NMR (400 MHz, Chloroform-d) δ 13.94 (s, 1H), 13.37 (s, 1H), 8.05 (d, J = 7.7 Hz, 1H), 7.78 (t, J = 8.1 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 5.52 (t, J = 4.5 Hz, 1H), 5.31 (dd, J = 4.5, 2.5 Hz, 1H), 5.16 (dd, J = 5.3, 1.8 Hz, 1H), 4.75 (s, 1H), 4.14 (qd, J = 6.5, 2.0 Hz, 1H), 4.09 (s, 3H), 4.01 (q, J = 5.3 Hz, 2H), 3.85 (dd, J = 5.6, 2.0 Hz, 1H), 3.47 - 3.37 (m, 1H), 3.35 (t, J = 5.3 Hz, 1H), 3.32 - 3.22 (m, 1H), 3.15 - 3.05 (m, 1H), 2.68 - 2.54 (m, 3H), 2.10 - 2.01 (m, 3H), 1.93 - 1.68 (m, 6H), 1.41 (d, J = 6.6 Hz, 3H).

[0115] Example 2

[0116] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-lH-pyrrol-l- yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7- methoxy-N-(2-(methylamino)ethyl)-6,l l-dioxo-l,2,3,4,6,l l-hexahydro- tetraphenyl-2-carboxamide (Compound DP-5)

[0117] Step one:

[0118] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,l l- dioxo-l,2,3,4,6,l l-hexahydro-tetraphenyl-2-carboxylic acid (Compound 2-1)

[0119] Doxorubicin hydrochloride (500 mg, 0.86 mmol) was dissolved in a mixture solvent of methanol (72 mL) and water (48 mL) at room temperature, then a solution of NaIO4(220.7 mg, 1.03 mmol) in water (24 mL) was added, the reaction was continued at this temperature for 1 h, the reaction was monitored by LC-MS. The solvent was removed under reduced pressure, the crude product was used directly for the next step.

[0120] Step two:

[0121] Synthesis of tert-butyl (2-((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydro-2-tetra-benzofuran-carbonyl)ethyl)(methyl)carbamate (Compound 2-2)

[0122] The crude product of the previous step was dissolved in DMF (30 mL) at room temperature, followed by the addition of N-Boc-N-methylethylenediamine (300 mg, 1.72 mmol), PyBOP (537.1 mg, 1.03 mmol) and N,N-diisopropylamine ethylamine (450 μL, 2.58 mmol) sequentially. The reaction was stirred for another 4 h, and the reaction was monitored by LC-MS. The solvent was evaporated, and the residue was dissolved in water (40 mL) and stirred well. The solution was extracted with ethyl acetate / isopropanol (3:1, 40 mL x 4) and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the residue was purified by C18 reverse phase column (acetonitrile-0.05% formic acid in water, 5%-95%) to give 180 mg of the brown red target product.

[0123] Step Three:

[0124] Synthesis of tert-butyl (2-((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H-pyrrol-1-yl)- 5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy- 6,11-dioxo-1,2,3,4,6,11-hexahydro-2-tetra-benzofuran-carbonyl)ethyl)(methyl) carbamate (Compound 2-3)

[0125] Compound 2-2 (180 mg, 0.26 mmol) was dissolved in DMF (2 mL) at room temperature, followed by the addition of 5-iodovaleraldehyde (1.38 g, 6.5 mmol) and N,N-diisopropyl ethylamine (137 μL, 0.78 mmol) sequentially. The reaction was stirred for another 10 min, and the reaction was monitored by LC-MS. The reaction was purified by C18 reverse phase column (acetonitrile-0.1% TFA in water, 5%-95%) to give 102 mg of the brown red title compound.

[0126] Step Four:

[0127] Synthesis of Compound DP-5

[0128] To a solution of compound 2-3 (102 mg, 0.14 mmol) in dichloromethane (8 ml) was added zinc bromide (438.8 mg, 1.95 mmol) in portions at room temperature. The reaction was monitored by LC-MS at room temperature for 18 h. Concentrated under reduced pressure, the residue was dissolved in 2 mL DMF and purified by HPLC prep (acetonitrile-0.05% formic acid in water, 15%-35%, 30 mL / min) to give 16 mg of the formate salt of the title compound as dark brown red.

[0129] LC-MS: 638.3 [M+H] +

[0130] 1 H NMR (400 MHz, Chloroform-d) δ 13.93 (s, 1H), 8.53 (s, 1H), 8.01 (d, J = 7.6 Hz, 1H), 7.93 - 7.84 (m, 1H), 7.75 (t, J = 8.1 Hz, 1H), 7.32 (d, J = 8.5 Hz, 1H), 5.53 (t, J = 4.6 Hz, 1H), 5.31 (d, J = 3.4 Hz, 1H), 5.16 (dd, J = 5.3, 1.7 Hz, 1H), 4.08 (tt, J = 6.7, 4.0 Hz, 2H), 4.03 (s, 3H), 3.89 (dd, J = 6.0, 1.9 Hz, 1H), 3.61 (t, J = 10.2 Hz, 3H), 3.38 - 3.33 (m, 1H), 3.22 - 3.03 (m, 5H), 3.02 - 2.92 (m, 2H), 2.61 (s, 4H), 2.42 - 2.31 (m, 3H), 2.14 - 2.04 (m, 2H), 1.94 - 1.70 (m, 2H), 1.35 (d, J = 6.5 Hz, 3H).

[0131] Example 3

[0132] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-lH-pyrrol-l-yl)-5- hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-N-(4- (methylamino)benzyl)-6,l l-dioxo-l,2,3,4,6,l l-hexahydro-tetraceno-2-carboxamide (Compound DP-30).

[0133] Step one:

[0134] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro- tetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)carbamate (Compound 3-1)

[0135] Doxorubicin hydrochloride (5.0 g, 8.62 mmol) was dissolved in Dioxane-H2O (150 mL, 1:2) mixed solvent at room temperature, the reaction solution was moved to an ice bath, NaHCO3(2.17 g, 25.86 mmol) was added, stirring was continued for 10 min, then Fmoc-Cl (2.45 g, 9.48 mmol) was added in batches, the reaction solution was moved to room temperature and stirring was continued for 1 h until the reaction was complete. The reaction was quenched with water (100 mL), extracted with dichloromethane (150 mL x 2), the organic phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated to obtain 6.6 g of the red target product which was directly used in the next step.

[0136] Step two:

[0137] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro-tetra- benzo-2-carboxylic acid (Compound 3-2)

[0138] Compound 3-1 (6.6 g, 8.62 mmol) was dissolved in THF-H2O (150 mL, 2:1) mixed solvent at room temperature, NaIO4(2.03 g, 9.48 mol) was added, stirring was continued at room temperature for 16 h until the reaction was complete. The reaction was quenched with water (100 mL), extracted twice with dichloromethane (150 mL x 2), the organic phases were combined, washed once with water (150 mL), the organic phase was dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol, 95:5) to finally obtain 6.4 g of the brownish red target product.

[0139] Step three:

[0140] Synthesis of 4-(aminomethyl)-N-methylaniline (synthesis of Compound 3-4)

[0141] To a solution of 4-methylaminobenzonitrile (500 mg, 3.8 mmol) in tetrahydrofuran (20 mL) was added lithium aluminum hydride (288 mg, 7.6 mmol) in portions at room temperature. The mixture was stirred at room temperature for another 4 h. LC-MS showed the reaction was completed. The reaction was quenched by the addition of saturated aqueous potassium sodium tartrate (100 mL). The mixture was extracted with dichloromethane (100 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (dichloromethane / methanol, 93:7) to give 350 mg of the target compound.

[0142] Step Four:

[0143] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)- 3,5,12-trihydroxy-10-methoxy-3-((4-(methylamino)benzyl)carbamoyl)-6,11-dioxo-1,2,3,4,6,11- hexahydro- tetraphenyl-1-yl)oxy)tetrahydro-2H-pyran-4-yl)carbamate (Compound 3-5)

[0144] To a solution of Compound 3-2 (717 mg, 0.95 mmol) in DMF (10 mL) was added HATU (363 mg, 0.95 mmol), Compound 3-4 (100 mg, 0.73 mmol) and N,N-diisopropylethylamine (285 mg, 2.20 mmol) in sequence at room temperature. The reaction was stirred at this temperature for another 4 h. LC-MS showed the reaction was completed. The reaction was diluted with methyl tert-butyl ether. The mixture was filtered to give 500 mg of the red target compound as a crude product.

[0145] Step Five:

[0146] Synthesis of (2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)-2,5,12-trihydroxy-7-methoxy-N-(4-(methylamino)benzyl)-6,11-dioxo-1,2,3,4,6,11- hexahydro- tetraphenalene-2-carboxamide (Compound 3-6)

[0147] To a solution of Compound 3-5 (500 mg, 0.58 mmol) in DMF (10 mL) was added diethylamine (841 mg, 11.50 mmol) at room temperature. The reaction was stirred at this temperature for another 0.5 h. LC-MS showed the reaction was completed. The reaction was diluted with methyl tert-butyl ether (5 mL). The mixture was purified by column chromatography on silica gel (dichloromethane / methanol, 82:18) to give 330 mg of the red target compound.

[0148] Step six:

[0149] Synthesis of compound DP-30

[0150] To a solution of compound 3-6 (330 mg, 0.51 mmol) in DMF (10 mL) was added 4-iodobutyraldehyde (757 mg, 3.82 mmol) and N,N-diisopropylethylamine (330 mg, 2.55 mmol) sequentially at room temperature, and the reaction was stirred at room temperature for another 16 h under nitrogen. LC-MS showed that the reaction was completed. To the reaction solution was added acetonitrile (5 mL), water (5 mL), acetic acid (0.2 mL), trifluoroacetic acid (0.4 mL), and the mixture was stirred at room temperature for 0.5 h. The reaction mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (dichloromethane / methanol, 85:15) to obtain 150 mg of the target compound as a crude product. The crude product was further separated and purified by HPLC preparation (acetonitrile-0.05% formic acid aqueous solution, 20%-40%, 30 mL / min) to obtain 11 mg of the red target compound.

[0151] LC-MS: 700.3 [M+H] + ;

[0152] 1 H NMR (400 MHz, Methanol-d4) δ 7.95 (dd, J = 23.4, 7.7 Hz, 1H), 7.83 (dt, J = 14.9, 8.1 Hz, 1H), 7.62 - 7.46 (m, 2H), 7.11 (d, J = 8.1 Hz, 4H), 6.64 - 6.53 (m, 4H), 5.50 (dd, J = 21.2, 13.7 Hz, 2H), 5.14 (s, 1H), 4.59 (s, 2H), 4.48 - 4.28 (m, 2H), 4.27 - 4.11 (m, 3H), 4.07 - 3.95 (m, 6H), 3.81 (d, J = 15.9 Hz, 1H), 3.60 (s, 1H), 3.16 (d, J = 4.6 Hz, 2H), 3.11 (s, 1H), 2.75 (d, J = 1.3 Hz, 3H).

[0153] Example 4

[0154] Synthesis of (2S,4S)-N-(3-aminobenzyl)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H-pyrrol-1-yl)-5- hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11- hexahydroterbenzofuran-2-carboxamide (compound DP-34)

[0155] Step one:

[0156] Synthesis of tert-butyl (4-((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydro-2-methyl-1H-tetrabenzoazepin-4-ylamino)methyl)benzoate (Compound 4-2)

[0157] Compound 3-2 (1.13 g, 1.50 mmol) was dissolved in DMF (10 mL) at room temperature, then tert-butyl (4-aminophenyl)carbamate (208.0 mg, 1.0 mmol), HATU (569.6 mg, 1.50 mmol) and N,N-diisopropylethylamine (0.5 mL, 3.0 mmol) were added, and stirring was continued at room temperature for 3 h until the reaction was complete. The reaction was quenched with water (10 mL), extracted with dichloromethane (30 mL x 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product which was used directly in the next step.

[0158] Step two:

[0159] Synthesis of tert-butyl (4-((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydro-2-methyl-1H-tetrabenzoazepin-4-ylamino)methyl)benzoate (Compound 4-2)

[0160] The crude product of compound 4-1 (900 mg, 0.96 mmol) was dissolved in DMF (10 mL) at room temperature, then diethylamine (3.0 mL, 28.7 mmol) was added, and stirring was continued at room temperature for 15 min until the reaction was complete. The solvent and diethylamine were removed under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol, 85:15) to give 660 mg of the red target compound.

[0161] Step three:

[0162] Synthesis of tert-butyl (4-((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H-pyrrol-1-yl)- 5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy- 6,11-dioxo-1,2,3,4,6,11-hexahydro-2-methyl-1H-tetrabenzoazepin-4-ylamino)methyl)benzoate (Compound 4-3)

[0163] Compound 4-2 (660 mg, 0.92 mmol) was dissolved in DMF (10 mL) at room temperature, followed by the addition of 4-iodobutyraldehyde (1.36 g, 6.88 mmol) and N,N- diisopropylethylamine (0.76 mL, 4.59 mmol) successively, and the reaction was stirred at room temperature for 24 h under nitrogen. The reaction was monitored by LC-MS. Acetonitrile (5 mL), water (5 mL), acetic acid (0.2 mL) and trifluoroacetic acid (0.4 mL) were added successively to the reaction solution, and the mixture was stirred at room temperature for 0.5 h. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol, 90:10) to give 300 mg of the red target compound.

[0164] Step four:

[0165] Synthesis of compound DP-34

[0166] Compound 4-3 (300 mg, 0.39 mmol) was dissolved in dichloromethane (15 mL) at room temperature, followed by the addition of zinc bromide (4.38 g, 19.4 mmol) in portions, and the reaction was stirred at room temperature for 2 h. The dichloromethane was removed under reduced pressure, acetonitrile (10 mL) was added to the residue, and the zinc bromide was removed by filtration. The filtrate was collected and purified by HPLC preparation (acetonitrile-0.05% formic acid aqueous solution, 20%-40%, 30 mL / min) to give 6.2 mg of the brown red title compound.

[0167] LC-MS: 672.5 [M+H] +

[0168] 1 H NMR (400 MHz, Methanol-d4) δ 8.53 (s, 1H), 8.06 - 7.94 (m, 1H), 7.85 (t, J = 8.1 Hz, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.32 (t, J = 8.4 Hz, 2H), 6.73 (d, J = 8.4 Hz, 2H), 5.60 - 5.43 (m, 1H), 5.24 (s, 2H), 4.58 (s, 3H), 4.33 - 4.24 (m, 1H), 4.04 (s, 3H), 3.53 - 3.42 (m, 1H), 3.21 (d, J = 9.8 Hz, 2H), 3.13 (d, J = 2.2 Hz, 1H), 2.75 - 2.62 (m, 1H), 2.43 (d, J = 10.5 Hz, 2H), 2.10 (d, J = 8.8 Hz, 1H), 1.78 (dd, J = 52.7, 6.0 Hz, 2H), 1.38 - 1.19 (m, 3H).

[0169] Example 5

[0170] Synthesis of (2S,4S)-N-(4-amino-3-methylphenyl)-4-(((2R,4S,5S,6S)-4-(2,3- dihydro-1 H-pyrrol-1 -yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12- trihydroxy-7-methoxy-6,11 -dioxo-1,2,3,4,6,11 -hexahydroterbenzofuran-2-carboxamide (Compound DP-37)

[0171] Step one:

[0172] Synthesis of tert-butyl (2-methyl-4-nitrophenyl)carbamate (Compound 5-2)

[0173] The 2-methyl-4-nitroaniline (1000 mg, 6.57 mmol) was dissolved in dichloromethane (20 mL) at room temperature, followed by the addition of triethylamine (2.73 mL, 19.72 mmol), DMAP (80.3 mg, 0.66 mmol) and di-tert-butyl dicarbonate (1.66 mL, 7.23 mmol) successively. The reaction was stirred for 6 h at room temperature, and LC-MS was used to monitor the reaction. The reaction was quenched by the addition of water (50 mL) and extracted with dichloromethane (50 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 97:3) to give 580 mg of the light yellow target product.

[0174] Step two:

[0175] Synthesis of tert-butyl (4-amino-2-methylphenyl)carbamate (Compound 5-3)

[0176] The compound 5-2 (580 mg, 2.30 mmol) was dissolved in a solution of ethanol / water (18 mL, v / v = 2:1) at room temperature, followed by the addition of ammonium chloride (614.9 mg, 11.50 mmol) and iron powder (642 mg, 11.50 mmol) successively. The reaction was stirred at 85 °C for 1 h, and LC-MS was used to monitor the reaction. The reaction was cooled to room temperature, diluted with ethanol (12 mL), filtered using celite, and the filtrate was collected and concentrated under reduced pressure. The residue was dissolved in water (50 mL), extracted with dichloromethane (50 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 87:13) to give 500 mg of the brown yellow target product.

[0177] Step three:

[0178] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-6-(((1S,3S)-3-((4-((tert- butoxycarbonyl)amino)-3-methylphenyl)carbamoyl)-3,5,12-trihydroxy-10- methoxy-6, 11 -dioxo- 1,2,3,4,6, 11 -hexahydro- tetraphen- 1 -yl)oxy)-3 -hydroxy-2- methyltetrahydro-2H-pyran-4-yl)carbamate (Compound 5-4)

[0179] Compound 3-2 (659.5 mg, 0.88 mmol) was dissolved in DMF (5 mL) at room temperature, and HATU (333.6 mg, 0.88 mmol) was added, and the reaction was stirred for 0.5 h. In another reaction flask, compound 5-3 (150 mg, 0.68 mmol) was dissolved in DMF (5 mL), and DMAP (107.2 mg, 0.88 mmol) was added, and the reaction was stirred for 0.5 h at room temperature. The reaction was added dropwise to the previous reaction. The reaction was stirred for 40 min at room temperature, and LC-MS indicated that the reaction was complete. Methyl tert-butyl ether (250 mL) was added to precipitate the product, which was filtered to give 600 mg of the crude product as a red solid.

[0180] Step Four:

[0181] Synthesis of tert-butyl (4-((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6, 11 -dioxo- 1,2,3,4,6, 11 -hexahydro- tetra-2-carboxamido)-2-methylphenyl)carbamate (Compound 5-5)

[0182] Compound 5-4 (600 mg, 0.63 mmol) was dissolved in DMF (10 mL) at room temperature, and diethylamine (921 mg, 12.60 mmol) was added, and the reaction was stirred for 0.5 h, and LC-MS indicated that the reaction was complete. The reaction was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol, 80:20) to give 110 mg of the red target product.

[0183] Step Five:

[0184] Synthesis of tert-butyl (4-((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-lH-pyrrol-l- yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy- 6, 11 -dioxo- 1,2,3,4,6, 11 -hexahydroterebenzyl-2-carbonyl)amino)methyl)phenyl)aminoformate (Compound 5-6)

[0185] Compound 5-5 (110 mg, 0.15 mmol) was dissolved in DMF (10 mL) at room temperature, then 4-iodobutyraldehyde (296.8 mg, 1.50 mmol) and N,N- diisopropylethylamine (0.12 mL, 0.75 mmol) were added successively, the reaction was stirred for 16 h, LC-MS detection showed the reaction was complete. Water (5 mL), acetic acid (0.05 mL), acetonitrile (20 mL) and trifluoroacetic acid (0.1 mL) were added to the reaction, stirred for 0.5 h, concentrated under reduced pressure, the residue was separated by silica gel column chromatography (dichloromethane / methanol, 90:10) to obtain 110 mg of the red target compound.

[0186] Step six:

[0187] Synthesis of compound DP-37

[0188] Compound 5-6 (100 mg, 0.13 mmol) was dissolved in dichloromethane (10 mL) at room temperature, then zinc bromide (1.43 g, 6.36 mmol) was added, the reaction was stirred for 1 h, LC-MS detection showed the reaction was complete. Concentrated under reduced pressure, the residue was slurried in acetonitrile (20 mL) for 10 min, filtered, the filtrate was collected and concentrated under reduced pressure again, the residue was separated and purified by HPLC preparation (acetonitrile-0.05% formic acid aqueous solution, 20%-40%, 30 mL / min) to obtain 8 mg of the red target compound.

[0189] LC-MS: 686.3 [M+H] + .

[0190] 1H NMR (400 MHz, Chloroform-d) δ 13.99 (s, 1H), 13.28 (s, 1H), 8.83 (s, 1H), 8.05 (d, J = 7.7 Hz, 1H), 7.78 (t, J = 8.1 Hz, 1H), 7.39 (dd, J = 5.6, 3.0 Hz, 2H), 7.28 (d, J = 2.2 Hz, 1H), 6.67 (d, J = 8.4 Hz, 1H), 5.57 (t, J = 4.8 Hz, 1H), 5.40 (d, J = 3.1 Hz, 1H), 5.33 (s, 1H), 5.17 (d, J = 5.1 Hz, 1H), 4.09 (s, 3H), 4.08 - 4.01 (m, 2H), 3.90 (dd, J = 5.9, 2.0 Hz, 1H), 3.33 (dd, J = 8.0, 4.7 Hz, 3H), 3.15 (s, 1H), 2.69 - 2.58 (m, 1H), 2.48 (d, J = 4.1 Hz, 2H), 2.19 (s, 3H), 2.14 - 2.05 (m, 1H), 1.99 - 1.79 (m, 2H), 1.79 - 1.69 (m, 2H), 1.35 (d, J = 6.5 Hz, 3H).

[0191] Example 6

[0192] Synthesis of (2S,4S)-N-(3-amino-4-methylbenzyl)-4-(((2R,4S,5S,6S)-4-(2,3- dihydro-1H-pyrrol-1-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12- trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydroretrabenzo-2-carboxamide (Compound DP-39)

[0193] Step one:

[0194] Synthesis of 5-(aminomethyl)-2-methylanilines compound (Compound 6-2)

[0195] At room temperature, 3-amino-4-methylbenzamide (1.00 g, 6.66 mmol) was dissolved in tetrahydrofuran (20 mL), lithium aluminum hydride tetrahydrofuran solution (6 mL, 15.00 mmol, 2.5 M) was added at 0 °C, then stirred at 60 °C for 2 h, LC-MS detected that the reaction was complete. Cooled to room temperature, water (0.5 mL), 15% sodium hydroxide aqueous solution (0.5 mL), water (1.5 mL) were added slowly to the reaction system, stirred for 0.5 h, then dried with anhydrous sodium sulfate, filtered, concentrated to give 0.9 g of brown crude product which was used directly in the next step.

[0196] Step two:

[0197] Synthesis of (9H-fluoren-9-yl)methyl (3-amino-4-methylbenzyl)carbamate (Compound 6-3)

[0198] Compound 6-2 (0.90 g, 6.61 mmol) was dissolved in N,N-dimethylformamide (12 mL) at room temperature, followed by the addition of 9-fluorenylmethyl N-succinimidyl carbonate (2.30 g, 6.82 mmol) and N,N-diisopropylethylamine (2.3 mL, 13.22 mmol). The reaction was stirred at room temperature for 2 h, and LC-MS indicated that the reaction was complete. The reaction was diluted with ethyl acetate (30 mL), and the organic phase was washed with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol, 95:5) to give 1.0 g of the white target compound.

[0199] Step Three:

[0200] Synthesis of (9H-fluoren-9-yl)methyl (3-((tert-butoxycarbonyl)amino)-4- methylbenzyl)carbamate (Compound 6-4):

[0201] Compound 6-3 (1.00 g, 2.79 mmol) was dissolved in tetrahydrofuran (35 mL) at room temperature, followed by the addition of di-tert-butyl dicarbonate (1.9 mL, 8.27 mmol) and a solution of sodium hydroxide (125 mg, 3.13 mmol) in water (7 mL). The reaction was stirred at 60 °C for 2 h, and TLC indicated that the reaction was complete. After the reaction was cooled to room temperature, it was diluted with ethyl acetate (50 mL), and the organic phase was washed with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol, 95:5) to give 1.3 g of the white target compound.

[0202] Step Four:

[0203] Synthesis of tert-butyl (5-(aminomethyl)-2-methylphenyl)carbamate (Compound 6-5)

[0204] Compound 6-4 (700 mg, 1.53 mmol) was dissolved in N,N-dimethylformamide (7 mL) at room temperature, followed by the addition of diethylamine (1.6 mL, 15.53 mmol). The reaction was stirred at room temperature for 1 h, and LC-MS indicated that the reaction was complete. The reaction was concentrated under reduced pressure, and the residue was swelled with petroleum ether to give 240 mg of the white product, which was used directly in the next step.

[0205] Step Five:

[0206] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-6-(((1S,3S)-3-((3-((tert- butoxycarbonyl)amino)-4-methylbenzyl)carbamoyl)-3,5,12-trihydroxy-10- methoxy-6, 11 -dioxo- 1,2,3,4,6, 11 -hexahydro- tetraphen- 1 -yl)oxy)-3 -hydroxy-2- methyltetrahydro-2H-pyran-4-yl)carbamate (Compound 6-6)

[0207] Compound 3-2 (500 mg, 0.67 mmol) and compound 6-5 (200 mg, 0.85 mmol) were dissolved in N,N-dimethylformamide (8 mL) at room temperature, followed by the addition of HATU (380 mg, 1.0 mmol) and N,N-diisopropylethylamine (0.23 mL, 1.32 mmol). The reaction was stirred at room temperature for 2 h, and LC-MS showed that the reaction was complete. The reaction was diluted with ethyl acetate (20 mL), and the organic phase was washed with water (20 mL) and saturated brine (20 mL) successively, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (dichloromethane / methanol, 95:5) to give 260 mg of the red target compound.

[0208] Step six:

[0209] Synthesis of tert-butyl (5-(((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6, 11 -dioxo- 1,2,3,4,6, 11 -hexahydro- tetra-2-carboxamido)methyl)-2-methylphenyl)carbamate (Compound 6-7)

[0210] Compound 6-6 (210 mg, 0.22 mmol) was dissolved in N,N-dimethylformamide (3 mL) at room temperature, followed by the addition of diethylamine (0.23 mL, 2.23 mmol). The reaction was stirred at room temperature for 1 h, and LC-MS showed that the reaction was complete. The reaction was filtered and directly purified by reverse phase column (acetonitrile-0.05% formic acid in water, 5%-95%) to give 70 mg of the red target compound.

[0211] Step seven:

[0212] Synthesis of tert-butyl (5-(((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-lH- pyrrol-l-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12- trihydroxy-7-methoxy-6, 11-dioxo-l,2,3,4,6, 11-hexahydroterebenzyl-2- carboxamido)methyl)-2-methylphenyl)carbamate (Compound 6-8)

[0213] Compound 6-7 (70 mg, 0.094 mmol) was dissolved in N,N-dimethylformamide (2 mL) at room temperature, followed by the addition of 4-iodobutyraldehyde (190 mg, 0.96 mmol) and diisopropylethylamine (0.05 mL, 0.29 mmol). The reaction was stirred at room temperature for 1 h, and LC-MS indicated that the reaction was complete. The reaction was filtered and directly purified by reverse phase column (acetonitrile-0.05% formic acid in water, 5%-95%) to give 40 mg of the red target compound.

[0214] Step Eight:

[0215] Synthesis of Compound DP-39:

[0216] Compound 6-8 (35 mg, 0.044 mmol) was dissolved in dichloromethane (8 mL) at room temperature, followed by the addition of zinc bromide (990 mg, 4.40 mmol). The reaction was stirred at room temperature for 3 h, and LC-MS indicated that the reaction was complete. The reaction was directly concentrated, slurried in acetonitrile (10 mL) for 10 min, filtered, and purified by HPLC (acetonitrile-0.05% formic acid in water, 10-40%, 25 mL / min) to give 10.8 mg of the red target compound.

[0217] LC-MS: [M+H] + 700.4

[0218] 1H NMR (400 MHz, DMSO-d6) δ 14.05 (s, 1H), 13.23 (s, 1H), 8.25 (t, J = 6.1 Hz, 1H), 7.90 (d, J = 4.4 Hz, 2H), 7.65 (q, J = 4.9, 4.2 Hz, 1H), 6.83 (d, J = 7.7 Hz, 1H), 6.49 (s, 1H), 6.37 (dd, J = 7.5, 1.4 Hz, 1H), 5.37 (s, 1H), 5.25 (t, J = 3.8 Hz, 1H), 5.04 - 4.91 (m, 2H), 4.75 (s, 2H), 4.24 - 4.05 (m, 3H), 3.98 (s, 3H), 3.73 (dd, J = 5.4, 1.8 Hz, 1H), 3.29 - 3.20 (d, J = 7.9 Hz, 2H), 3.08 (d, J = 18.3 Hz, 1H), 2.99 - 2.88 (m, 2H), 2.58 - 2.51 (m, 1H), 2.31 (dd, J = 14.4, 5.3 Hz, 1H), 2.17 (d, J = 11.7 Hz, 1H), 2.01 (s, 3H), 1.93 (dd, J = 7.6, 5.3 Hz, 1H), 1.77 - 1.50 (m, 4H), 1.21 - 1.08 (m, 3H).

[0219] Example 7

[0220] Synthesis of (2S,4S)-N-(4-amino-3-bromophenyl)-4-(((2R,4S,5S,6S)-4-(2,3- dihydro-1H-pyrrol-1-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12- trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydroretrabenzo-2-carboxamide (Compound DP-42)

[0221] Step one:

[0222] Synthesis of tert-butyl (2-bromo-4-nitrophenyl)carbamate (Compound 7-2)

[0223] To a solution of 2-bromo-4-nitroaniline (3.0 g, 13.82 mmol) in dichloromethane (60 mL) was added triethylamine (4.2 g, 41.47 mmol) and di-tert-butyl dicarbonate (6.03 g, 27.65 mmol) sequentially at room temperature. The mixture was stirred for 18 h. LC-MS showed the reaction was completed. The reaction was quenched by adding water (100 mL) and extracted with dichloromethane (100 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 99:1) to give 1.5 g of the white target product.

[0224] Step two:

[0225] Synthesis of tert-butyl (4-amino-2-bromophenyl)carbamate (compound 7-3)

[0226] To a solution of compound 7-2 (1.0 g, 3.15 mmol) in ethanol / water (30 mL, v / v = 2:1) was added ammonium chloride (1.69 g, 31.53 mmol) and iron powder (1.76 g, 31.53 mmol) sequentially at room temperature. The mixture was stirred at 85 °C for 1.5 h. LC-MS indicated that the reaction was complete. The reaction was cooled to room temperature, and the iron powder was removed by filtration using celite. The filtrate was collected and concentrated to remove ethanol to give an aqueous phase, which was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated by column chromatography on silica gel (petroleum ether / ethyl acetate, 91:9) to give 690 mg of the white target compound.

[0227] Step three:

[0228] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-6-(((1S,3S)-3-((3-bromo-4- ((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-3,5,12-trihydroxy-10-methoxy-6,11- dioxo-1,2,3,4,6,11-hexahydro-4-terphenyl-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H- pyran-4-yl)carbamate (compound 7-4)

[0229] To a solution of compound 3-2 (510 mg, 0.53 mmol) in DMF (10 mL) was added HATU (255 mg, 0.68 mmol), compound 7-3 (100 mg, 0.73 mmol), and N,N- diisopropylethylamine (285 mg, 2.20 mmol) sequentially at room temperature. The reaction was stirred for 2 h. LC-MS indicated that the reaction was complete. Acetonitrile (50 mL) was added to the reaction, which was then concentrated. This was repeated three times. The remaining DMF solution was crystallized by adding methyl tert-butyl ether. Filtration gave 341 mg of the red crude target compound, which was used directly in the next reaction.

[0230] Step four:

[0231] Synthesis of tert-butyl (4-((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydroterbenzofuran-2-carboxamido)-2-bromophenyl)carbamate (Compound 7-5)

[0232] To a solution of compound 7-4 (341 mg, 0.33 mmol) in DMF (10 mL) was added diethylamine (997 mg, 6.68 mmol) at room temperature. The reaction was stirred for 0.5 h at room temperature until LC-MS indicated the reaction was complete. The reaction was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane / methanol, 80:20) to give 210 mg of the red target compound.

[0233] Step five:

[0234] Synthesis of tert-butyl (2-bromo-4-((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H- pyrrol-1-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy- 7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydroterbenzofuran-2-carboxamido)phenyl) carbamate (Compound 7-6)

[0235] To a solution of compound 7-5 (210 mg, 0.26 mmol) in DMF (10 mL) was added 4-iodobutyraldehyde (391 mg, 1.97 mmol) followed by N,N- diisopropylethylamine (171 mg, 1.32 mmol) at room temperature. The reaction was stirred for 16 h at room temperature until LC-MS indicated the reaction was complete. To the reaction was added acetonitrile and water (5 mL each) followed by acetic acid (0.1 mL) and trifluoroacetic acid (0.1 mL). The reaction was stirred for 0.5 h at room temperature. The reaction was concentrated. To the concentrate was added methyl tert-butyl ether (300 mL) dropwise. The red solid was collected by filtration to give 182 mg of the red target compound.

[0236] Step six:

[0237] Synthesis of compound DP-42

[0238] To a solution of compound 7-6 (170 mg, 0.20 mmol) in dichloromethane (30 mL) was added zinc bromide (2.25 g, 10 mmol) at room temperature, the mixture was stirred at room temperature for 1 h, LC-MS showed the reaction was complete. The residue was concentrated under reduced pressure, acetonitrile (30 mL) was added to the residue, slurried for 10 min, filtered, the filtrate was collected and concentrated to give a crude product, which was separated by HPLC preparation (acetonitrile-0.05% formic acid in water, 20%-40%, 30 mL / min) to give 4.2 mg of the red target compound.

[0239] LC-MS: 750.2 [M+H] + .

[0240] 1 H NMR (400 MHz, Chloroform-d) δ 13.99 (s, 1H), 13.28 (s, 1H), 8.85 (s, 1H), 8.05 (d, J = 7.6 Hz, 1H), 7.84-7.74 (m, 2H), 7.38 (dd, J = 10.6, 7.6 Hz, 2H), 6.77 (d, J = 8.6 Hz, 1H), 5.58 (s, 1H), 5.43-5.34 (m, 2H), 5.17 (s, 1H), 4.09 (s, 4H), 4.03 (d, J = 6.9 Hz, 3H), 3.91 (d, J = 5.9 Hz, 1H), 3.31 (d, J = 4.1 Hz, 2H), 3.16 (s, 1H), 2.68-2.59 (m, 1H), 2.47 (d, J = 4.6 Hz, 2H), 2.18-2.01 (m, 1H), 2.01-1.64 (m, 3H), 1.36 (d, J = 6.6 Hz, 3H).

[0241] Example 8

[0242] Synthesis of (2S,4S)-N-(3-amino-4-fluorobenzyl)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H- pyrrol-1-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7- methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydroretrabenzo-2-carboxamide (Compound DP-50)

[0243] Step one:

[0244] Synthesis of tert-butyl (5-cyano-2-fluorophenyl)formamide (Compound 8-2)

[0245] To a solution of compound 8-2 (1.5 g, 6.35 mmol) in MeOH (10 mL) was added anhydrous NiCl2(279.0 mg, 1.27 mmol) at room temperature, followed by the addition of NaBH4(1.20 g, 31.75 mmol) in portions at ice bath. The reaction was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction was quenched by the addition of H2O (20 mL) at ice bath. The mixture was extracted with dichloromethane (80 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (dichloromethane / methanol, 95:5) to give 450 mg of the brown black target compound.

[0246] Step two:

[0247] Synthesis of tert-butyl (5-(((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11- hexahydroterebenzepine-2-carboxamido)methyl)-2-fluorophenyl)carbamate (compound 8-4)

[0248] To a solution of compound 8-2 (1.5 g, 6.35 mmol) in MeOH (10 mL) was added anhydrous NiCl2(279.0 mg, 1.27 mmol) at room temperature, followed by the addition of NaBH4(1.20 g, 31.75 mmol) in portions at ice bath. The reaction was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction was quenched by the addition of H2O (20 mL) at ice bath. The mixture was extracted with dichloromethane (80 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (dichloromethane / methanol, 95:5) to give 450 mg of the brown black target compound.

[0249] Step three:

[0250] Synthesis of tert-butyl (5-(((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11- hexahydroterebenzepine-2-carboxamido)methyl)-2-fluorophenyl)carbamate (compound 8-4)

[0251] Compound 2-1 (1.15 g, 2.16 mmol) was dissolved in DMF (15 mL) at room temperature, then HATU (822.9 mg, 2.16 mmol) was added, and the mixture was stirred for 5 min. Compound 8-3 (400 mg, 1.67 mmol) and N,N-diisopropylethylamine (0.83 mL, 5.0 mmol) were added to the reaction mixture in turn. The reaction mixture was stirred at room temperature for 1 h, and LC-MS detection showed that the reaction was complete. The reaction mixture was diluted with methyl tert-butyl ether (10 mL), and separated by silica gel column chromatography (dichloromethane / methanol, 80:20) to obtain 434 mg of the red target product.

[0252] Step Four:

[0253] Synthesis of tert-butyl (5-(((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-1H-pyrrol-1-yl)-5- hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydroretrabenzo-2-carboxamido)methyl)-2-fluorophenyl)carbamate (Compound 8-5)

[0254] Compound 8-4 (420 mg, 0.56 mmol) was dissolved in DMF (10 mL) at room temperature, and 4-iodobutyraldehyde (829.7 mg, 4.19 mmol) and N,N-diisopropylethylamine (0.46 mL, 2.79 mmol) were added in turn, and the mixture was replaced with nitrogen three times. The reaction mixture was stirred at room temperature for 16 h, and LC-MS detection showed that the reaction was complete. Acetonitrile and water (5 mL each) were added to the reaction mixture, and acetic acid (0.1 mL) and trifluoroacetic acid (0.1 mL) were added in turn, and the mixture was stirred at room temperature for 0.5 h. The mixture was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (dichloromethane / methanol, 90:10) to obtain 342 mg of the red target compound.

[0255] Step Five:

[0256] Synthesis of Compound DP-50

[0257] Compound 8-5 (340 mg, 0.42 mmol) was dissolved in dichloromethane (10 mL) at room temperature, and zinc bromide (4.76 g, 21.15 mmol) was added in portions. The mixture was stirred at room temperature for 1 h, and LC-MS detection showed that the reaction was complete. The dichloromethane was removed by concentration, and acetonitrile (15 mL) was added. The mixture was slurried for 10 min, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by HPLC preparation (acetonitrile-0.05% formic acid aqueous solution, 20%-40%, 30 mL / min) to obtain 6 mg of the red target compound.

[0258] LC-MS: 704.5 [M+H] + .

[0259] 1 H NMR (400 MHz, Chloroform-d) δ 13.96 (s, 1H), 13.30 (s, 1H), 8.05 (d, J = 7.6 Hz, 1H), 7.78 (t, J = 8.1 Hz, 1H), 7.45 (d, J = 6.1 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 6.95 (dd, J = 10.9, 8.2 Hz, 1H), 6.75 (dd, J = 8.5, 2.2 Hz, 1H), 6.63 (dd, J = 8.8, 4.9 Hz, 1H), 5.52 (t, J = 4.7 Hz, 1H), 5.35 (d, J = 3.1 Hz, 1H), 5.18 - 5.11 (m, 2H), 4.49 - 4.30 (m, 2H), 4.09 (s, 4H), 3.98 (d, J = 7.1 Hz, 1H), 3.74 (s, 3H), 3.38 - 3.21 (m, 4H), 3.13 (s, 1H), 2.61 (d, J = 8.3 Hz, 1H), 2.43 (d, J = 4.5 Hz, 2H), 2.07 (d, J = 12.4 Hz, 1H), 1.97 - 1.78 (m, 3H), 1.30 (d, J = 6.6 Hz, 3H).

[0260] Example 9

[0261] Synthesis of (2S,4S)-N-(4-amino-3,5-dimethylbenzyl)-4-(((2R,4S,5S,6S)-4-(2,3- dihydro-1H-pyrrol-1-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12- trihydroxy-7-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro- tetra-benzo-2-carboxamide (Compound DP-51)

[0262] Step one:

[0263] Synthesis of 4-(aminomethyl)-2,6-dimethylaniline (Compound 9-2)

[0264] ​Compound 9-2 (100 mg, 0.67 mmol) was dissolved in acetonitrile (5 mL) at room temperature, then 9-fluorenylmethyl N-succinimidyl carbonate (225 mg, 0.67 mmol) and triethylamine (0.09 mL, 0.67 mmol) were added successively. The reaction was stirred at room temperature for 1 h, then LC-MS showed the reaction was completed. Water (5 mL) was added to the reaction, and the mixture was extracted with ethyl acetate (10 mL x 2). The organic phase was combined, dried, concentrated, and the residue was purified by silica gel column chromatography (methanol / dichloromethane, 1 :20) to give 100 mg of the white target compound.

[0265] Step two:

[0266] Synthesis of (9H-fluoren-9-yl)methyl (4-amino-3,5-dimethylbenzyl)carbamate (Compound 9-3)

[0267] Compound 9-2 (100 mg, 0.67 mmol) was dissolved in acetonitrile (5 mL) at room temperature, then 9-fluorenylmethyl N-succinimidyl carbonate (225 mg, 0.67 mmol) and triethylamine (0.09 mL, 0.67 mmol) were added successively. The reaction was stirred at room temperature for 1 h, then LC-MS showed the reaction was completed. Water (5 mL) was added to the reaction, and the mixture was extracted with ethyl acetate (10 mL x 2). The organic phase was combined, dried, concentrated, and the residue was purified by silica gel column chromatography (methanol / dichloromethane, 1 :20) to give 100 mg of the white target compound.

[0268] Step three:

[0269] Synthesis of (9H-fluoren-9-yl)methyl (4-((tert-butoxycarbonyl)amino)-3,5- dimethylbenzyl)carbamate (Compound 9-4)

[0270] Compound 9-3 (50 mg, 0.134 mmol) was dissolved in tetrahydrofuran (6 mL) at room temperature, then a solution of sodium hydroxide (5.91 mg, 0.15 mmol) in water (6 mL) and di-tert-butyl dicarbonate (88 mg, 0.40 mmol) were added successively. The mixture was heated at 70 °C for 4 h, and TLC showed the reaction was completed. The reaction was cooled to room temperature, diluted with water (6 mL), and extracted with ethyl acetate (10 mL x 2). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate, 80:20) to give 20 mg of the red target compound.

[0271] Step four:

[0272] Synthesis of tert-butyl (4-aminomethyl)-2,6-dimethylphenyl)carbamate (Compound 9-5)

[0273] Compound 9-4 (270 mg, 0.57 mmol) was dissolved in N,N-dimethylformamide (3 mL) at room temperature, diethylamine (0.88 mL, 8.57 mmol) was added, the reaction was stirred at room temperature for 1 h, LC-MS showed the reaction was complete. The reaction was directly purified by C18 reverse phase column (acetonitrile-0.05% formic acid in water, 5%-95%) to give 120 mg of the white target compound.

[0274] Step five:

[0275] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-6-(((1S,3S)-3-((4-((tert- butoxycarbonyl)amino)-3,5-dimethylbenzyl)carbamoyl)-3,5,12-trihydroxy-10- methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro-4-azatetrabenzo-2-yl)oxy)-3- hydroxy-2-methyltetrahydro-2H-pyran-4-yl)carbamate (Compound 9-6)

[0276] Compound 3-2 (300 mg, 0.40 mmol) and compound 9-5 (100 mg, 0.40 mmol) were dissolved in N,N-dimethylformamide (10 mL) at room temperature, HATU (167 mg, 0.44 mmol) and N,N-diisopropylethylamine (0.20 mL, 1.20 mmol) were added successively, the reaction was stirred at room temperature for 1 h, LC-MS showed the reaction was complete. The reaction was directly purified by C18 reverse phase column (acetonitrile-0.05% formic acid in water, 5%-95%) to give 100 mg of the red target compound.

[0277] Step six:

[0278] Synthesis of tert-butyl (4-(((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6,11-dioxo- 1,2,3,4,6,11-hexahydro-4-azatetrabenzo-2-carboxamido)methyl)-2,6-dimethyl- phenyl)carbamate (Compound 9-7)

[0279] Compound 9-6 (237 mg, 0.24 mmol) was dissolved in N,N-dimethylformamide (6 mL) at room temperature, diethylamine (0.37 mL, 3.61 mmol) was added, the reaction was stirred at room temperature for 1 h. The reaction was directly purified by C18 reverse phase column (acetonitrile-0.05% formic acid in water, 5%-95%) to give 90 mg of the red target compound.

[0280] Step seven:

[0281] Synthesis of tert-butyl (4-(((2S,4S)-4-(((2R,4S,5S,6S)-4-(2,3-dihydro-lH-pyrrol-l- yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7- methoxy-6, 11-dioxo-l,2,3,4,6, 11-hexahydroterebenzyl-2-carbonyl)amino)methyl)- 2,6-dimethylphenyl)carbamate (Compound 9-8)

[0282] Compound 9-7 (90 mg, 0.12 mmol) was dissolved in N,N-dimethylformamide (5 mL) at room temperature, then 4-iodobutyraldehyde (351 mg, 1.77 mmol) and N,N- diisopropylethylamine (0.11 mL, 0.70 mmol) were added, and the reaction was replaced with nitrogen three times. The reaction was stirred at room temperature for 16 h, and LC-MS showed that the reaction was complete. Acetonitrile and water (2.5 mL each) were added to the reaction, followed by acetic acid (0.05 mL) and trifluoroacetic acid (0.05 mL), and the mixture was stirred at room temperature for 0.5 h. The solvent was removed under reduced pressure, and the residue was purified by C18 reverse phase column (acetonitrile-0.05% formic acid aqueous solution, 5%-95%) to give 50 mg of the red target compound.

[0283] Step eight:

[0284] Synthesis of compound DP-51:

[0285] Compound 9-8 (50 mg, 0.061 mmol) was dissolved in dichloromethane (2 mL) at room temperature, then zinc bromide (692 mg, 3.07 mmol) was added, and the reaction was stirred at room temperature for 3 h, and LC-MS showed that the reaction was complete. The dichloromethane was removed by concentration, then acetonitrile (5 mL) was added, and the mixture was stirred for 10 min, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by HPLC preparation (acetonitrile-0.05% formic acid aqueous solution, 10-40%, 25 mL / min) to give 5.6 mg of the red target compound.

[0286] LC-MS: [M+H] + : 714.3

[0287] 1H NMR (400 MHz, DMSO-d6) δ 14.06 (s, 1H), 13.26 (s, 1H), 8.17 - 8.06 (m, 1H), 7.98 - 7.85 (m, 2H), 7.71 - 7.57 (m, 1H), 6.70 (s, 2H), 5.36 (s, 1H), 5.25 (t, J = 3.9 Hz, 1H), 5.01 (d, J = 3.6 Hz, 1H), 4.99 - 4.88 (m, 1H), 4.43 (s, 2H), 4.19 - 4.09 (m, 2H), 4.06 (dd, J = 14.2, 5.7 Hz, 1H), 3.99 (s, 3H), 3.73 (d, J = 7.2 Hz, 1H), 3.26 (t, J = 7.3 Hz, 1H), 3.09 (d, J = 18.2 Hz, 1H), 2.99 - 2.86 (m, 2H), 2.36 - 2.26 (m, 1H), 2.16 (s, 1H), 2.04 (s, 6H), 1.97-1.88 (m, 1H), 1.75 - 1.50 (m, 4H), 1.15 (d, J = 6.6 Hz, 3H).

[0288] Example 10

[0289] Synthesis of (2S,4S)-N-(3-aminobenzyl)-4-(((2R,4S,5S,6S)-4-(3,4- dihydropyridin-l(2H)-yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12- trihydroxy-7-methoxy-6, 11-dioxo-l,2,3,4,6, 11-hexahydro- tetra-benz [c, d] benzofuran-7- carboxamide (Compound DP-80)

[0290] Step one:

[0291] Synthesis of 5-iodopentanal (Compound 10-2):

[0292] The 5-iodopentan-l-ol (500 mg, 2.34 mmol) was dissolved in dichloromethane (5 mL) at room temperature, then Dess-Martin oxidant (1.49 g, 3.50 mmol) was added to the solution under ice bath, after 10 min, the reaction was allowed to warm to room temperature for 3 h, LC-MS showed the reaction was complete. The reaction was concentrated, the residue was separated by silica gel column chromatography (petroleum ether / ethyl acetate, 90: 10) to give 208 mg of colorless target compound.

[0293] Step two:

[0294] Synthesis of (9H-fluoren-9-yl)methyl ((2S,3S,4S,6R)-6-(((1S,3S)-3-((3- ((tert-butoxycarbonyl)amino)benzyl)carbamoyl)-3,5,12-trihydroxy-10- methoxy-6, 11-dioxo-1,2,3,4,6, 11-hexahydro- tetraphen-1-yl)oxy)-3-hydroxy-2-methyltetrahydro-2H-pyran-4-yl)carbamate (Compound 10-3)

[0295] Compound 3-2 (600 mg, 0.80 mmol) was dissolved in DMF (10 mL) at room temperature, followed by the addition of 3-(Boc-amino)benzylamine (177 mg, 0.80 mmol), N,N-diisopropylethylamine (206 mg, 1.60 mmol) and HATU (455.0 mg, 1.2 mmol). The reaction was stirred at room temperature for 1 h, and LC-MS indicated that the reaction was complete. The reaction was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (dichloromethane / methanol, 90:10) to give 720 mg of the red target product.

[0296] Step Three:

[0297] Synthesis of tert-butyl (3-(((2S,4S)-4-(((2R,4S,5S,6S)-4-amino-5-hydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7-methoxy-6, 11-dioxo- 1,2,3,4,6, 11-hexahydro- tetraphen-2-carboxamido)methyl)phenyl)carbamate (Compound 10-4)

[0298] Compound 10-3 (720 mg, 0.75 mmol) was dissolved in DMF (8 mL) at room temperature, followed by the addition of diethylamine (0.8 mL, 7.78 mmol). The reaction was stirred at room temperature for 0.5 h, and LC-MS indicated that the reaction was complete. The reaction was filtered, and the filtrate was purified by C18 reverse phase column (acetonitrile-0.1% trifluoroacetic acid in water, 5%-95%) to give 260 mg of the red target compound.

[0299] Step Four:

[0300] Synthesis of tert-butyl (3-(((2S,4S)-4-(((2R,4S,5S,6S)-4-(3,4-dihydropyridin-1(2H)- yl)-5-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2,5,12-trihydroxy-7- methoxy-6, 11-dioxo-1,2,3,4,6, 11-hexahydro- tetraphen-2-carboxamido)methyl)phenyl)carbamate (Compound 10-5):

[0301] Compound 10-4 (110 mg, 0.15 mmol) was dissolved in DMF (2 mL) at room temperature, then 5-iodovaleraldehyde (318 mg, 1.50 mmol) and N,N- diisopropylethylamine (58 mg, 0.45 mmol) were added. The reaction was stirred at room temperature for 0.5 h, and LC-MS showed the reaction was complete. The reaction was separated and purified by C18 reverse phase column (acetonitrile-0.01% formic acid in water, 5%-95%) to give 70 mg of the red target compound.

[0302] Step five:

[0303] Synthesis of compound DP-80:

[0304] Compound 10-5 (70 mg, 0.088 mmol) was dissolved in dichloromethane (2 mL) at room temperature, then zinc bromide (197 mg, 0.88 mmol) was added. The reaction was stirred at room temperature for 1 h, and LC-MS showed the reaction was complete. The dichloromethane was removed by concentration, then acetonitrile (4 mL) was added, and the mixture was slurried for 10 min, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by HPLC preparative (acetonitrile-0.05% formic acid in water, 20%-40%, 30 mL / min) to give 6 mg of the red target compound.

[0305] LC-MS: 700.6 [M+H] +

[0306] 1 H NMR (400 MHz, DMSO-d6) δ 14.04 (d, J = 23.1 Hz, 1H), 13.25 (s, 1H), 8.32 (t, J = 6.0 Hz, 1H), 7.98 - 7.87 (m, 2H), 7.72 - 7.60 (m, 1H), 6.93 (td, J = 7.7, 2.6 Hz, 1H), 6.43 (dd, J = 18.5, 7.9 Hz, 3H), 5.36 (d, J = 30.4 Hz, 1H), 5.27 (t, J = 7.0 Hz, 1H), 5.07 (s, 1H), 5.00 (s, 2H), 4.26 - 4.10 (m, 4H), 3.99 (s, 3H), 3.07 (d, J = 18.4 Hz, 2H), 3.01 - 2.91 (m, 3H), 2.25 - 2.07 (m, 2H), 1.96 - 1.72 (m, 3H), 1.70 - 1.56 (m, 2H), 1.26 (d, J = 21.4 Hz, 2H), 1.14 (dd, J = 22.8, 6.6 Hz, 3H).

[0307] Using the similar method described above, the following compounds were prepared.

[0308] Table 1

[0309] Cell killing assay of anthracene compounds

[0310] The purpose of this experiment is to detect the in vitro cell proliferation inhibition activity of representative compounds of the present application and control load molecules Dxd, Exatecan and MMAE on MDA-MB-231 (human breast cancer cells, Nanjing Kebai), NCI-H322 (human lung cancer cells, Nanjing Kebai), HCT-116 (human colorectal cancer cells, Nanjing Kebai), Calu-6 (human lung cancer cells, Nanjing Kebai), JIMT-1 (human breast cancer cells, Nanjing Kebai).

[0311] Experimental apparatus:

[0312] 96-well transparent flat-bottom white plate (Corning, #3610)

[0313] RPMI1640 medium (Gibco, #A10491-01)

[0314] DMEM medium (Gibco, #11995065)

[0315] Fetal Bovine Serum (Sigma-Aldrich, #F8687-500ML)

[0316] Luminescent Cell Viability Assay (Promega, #G7572)

[0317] Experimental method:

[0318] Human breast cancer cell MDA-MB-231, human lung cancer cell NCI-H322, human colorectal cancer cell HCT-116, human lung cancer cell Calu-6 and human breast cancer cell JIMT-1 were washed with PBS, and then trypsinized for about 3-10 minutes with 0.25% Trypsin-EDTA, and then terminated with complete medium, centrifuged at 1000 rpm for 3 minutes, discarded the supernatant, resuspended the cells with complete cell culture medium, counted with a cell counter, and adjusted the cells to the desired density, and then added 90.0 μL of cell suspension (3000 cells per well) to each well, and then placed the cell plate in a 37℃, 5% CO2 cell incubator for incubation overnight.

[0319] In the experiment, a DMSO control group and a test sample group were set up. The cell plate was taken out of the incubator to observe the cell adhesion state, and after the cells adhered, 10.00 μL of sample was added to each well (with a final concentration of 10 μM, 5-fold dilution, 9 concentrations), and then gently shaken and placed in a 37℃, 5% CO2 incubator for incubation.

[0320] After 3 days of incubation, 10.0 μL / well of Cell Counting Kit-8 (TargetMol, Catalog No.: C0005) working solution was added, and then the incubation in the cell incubator was continued for 0.5-4 hours. After taking out, shaking for 2 minutes, and then recording the absorbance in a plate reader (SpectraMax Paradigm).

[0321] Cell proliferation inhibition rate calculation formula: Cell proliferation inhibition rate % = 100% - (OD 化合物 -OD 空白 ) / (OD 内标 -OD 空白 ) x 100%.

[0322] Data analysis: plot the Log value of the sample concentration as the horizontal coordinate and the Inhibition % as the vertical coordinate, and perform Nonlinear regression (curve fit) analysis on the data to obtain the IC 50 value of each test sample.

[0323] Table 2 In vitro cell proliferation inhibition activity of human breast cancer cell MDA-MB-231, human lung cancer cell NCI-H322, and human colorectal cancer cell HCT-116

[0324] Table 3 In vitro cell proliferation inhibition activity of human lung cancer cell Calu-6

[0325] Table 4 In vitro cell proliferation inhibition activity of human breast cancer cell JIMT-1

[0326] Conclusion: As shown in Table 2, the compounds of the present application have moderate in vitro cell proliferation inhibitory activity.

[0327] The structure of the control load Dxd is as follows:

[0328] The structure of the control load Exatecan is as follows:

[0329] The structure of the control load MMAE is as follows:

Claims

1. A compound represented by Formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: ###00001### (I) wherein, R1is selected from H, hydroxy, C 1-6 alkoxy; R2is selected from O or NH; R3is selected from C 1-6 alkyl, hydroxyC 1-6 alkyl, -C(O)C 1-6 alkyl, -C(O)NR n1 R n2 ; R n1 and R n2 Each is independently selected from H, and arbitrarily selected by one or more R. n2-1 Replacement C 1-6 Alkyl, optionally with one or more R n2-2 Substituted 3- to 8-membered cycloalkyl groups, optionally with one or more R n2-3 Replaces the 4 to 10 element bridge ring base, optionally with one or more R n2-4 The substituted 3 to 8-membered heterocyclic group, optionally replaced by one or more R n2-5 The substituted 5 to 10 heteroaryl groups, optionally replaced by one or more R n2-6 The 6-10 aryl group that is replaced, of which, Each R n2-1 R n2-2 R n2-3 R n2-4 R n2-5 and R n2-6 Each is independently selected from hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogen, amino, hydroxy C 1-6 Alkyl, -NH-C 1-6 Alkyl, C 1-6 Alkyl-NH-C 1-6 Alkyl, nitro, cyano, halogenated C 1-6 Alkyl, ynyl, alkenyl, optionally with one or more R n2-1-1 The substituted 3 to 8-membered heterocyclic group, optionally replaced by one or more R n2-1-2 Substituted 3- to 8-membered cycloalkyl groups, optionally with one or more R n2-1-3 The substituted 6-10 aryl group and optionally one or more R n2-1-4 Substituted 5- to 10-membered heteroaryl groups; each R n2-1-1 , R n2-1-2 , R n2-1-3 , R n2-1-4 is independently selected from halogen, C 1-6 alkyl, amino, -NH-Ci-6alkyl, C 1-6 alkyl-NH2, nitro, cyano and halogenated C 1-6 alkyl; or, R n1 and R n2 together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl group, which is optionally substituted with C 1-6 alkyl, C 1-6 alkoxy, amino, hydroxy, -NH-C 1-6 alkyl, C 1-6 alkyl-NH-C 1-6 alkyl, hydroxyC 1-6 alkyl, nitro, cyano, trifluoromethyl, difluoromethyl, monofluoromethyl, trichloromethyl, alkynyl, alkenyl; R4and R5together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocycloalkenyl group.

2. The compound of Formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, which is selected from Formula (IA): ###0001### (IA) ​ wherein R1, R2, R3, R4, R5are as defined in claim 1.

3. The compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1 or 2, wherein R1is selected from -OCH3.

4. The compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-3, wherein R2is selected from O.

5. The compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-4, which satisfies one or more of the following conditions: (1) R3is selected from the group consisting of hydroxyC 1-6 alkyl and -C(O)NR n1 R n2 ; (2) R n1 selected from H and C 1-6 alkyl; preferably H; (3) R n2 selected from the group consisting of C n2-1 substituted C 1-6 alkyl, optionally substituted by one or more R n2-2 substituted 3- to 8-membered cycloalkyl, optionally substituted by one or more R n2-3 substituted 4- to 10-membered bridged ring group, 3- to 8-membered heterocyclyl, and 6- to 10-membered aryl, each of which is optionally substituted by one or more R n2-6 substituted 4- to 10-membered bridged ring group, 3- to 8-membered heterocyclyl, and 6- to 10-membered aryl, each of which is optionally substituted by one or more R (4) each R n2-1 each independently selected from the group consisting of hydroxyl, amino, -NH-C 1-6 alkyl, 3- to 8-membered heterocyclyl, 3- to 8-membered cycloalkyl, 6- to 10-membered aryl, and 5- to 10-membered heteroaryl, each optionally substituted with one or more R n2- 1-1 substituted 3- to 8-membered heterocyclyl, 3- to 8-membered cycloalkyl, 6- to 10-membered aryl, and 5- to 10-membered heteroaryl, each optionally substituted with one or more R n2-1-2 substituted 3- to 8-membered cycloalkyl, 6- to 10-membered aryl, and 5- to 10-membered heteroaryl, each optionally substituted with one or more R n2- 1-3 substituted 6- to 10-membered aryl; (5) each R is independently selected from the group consisting of halogen and C1-6alkyl; n2-1-1 each R is independently selected from the group consisting of halogen and C1-6alkyl; 1-6 alkyl; (6) each R n2-1-2 each independently amino; (7) each R n2-1-3 each independently selected from halogen, C 1-6 alkyl, amino, -NH-C 1-6 alkyl, C 1-6 alkyl-NH2, nitro, cyano and haloC 1-6 alkyl; (8) each R n2-3 and R n2-5 each independently is amino; (9) each R n2-4 each independently selected from amino, C 1-6 alkyl-NH2, and C 1-6 alkyl; (10) each R n2-6 each independently selected from the group consisting of halogen, C 1-6 alkyl, amino, C 1-6 alkyl-NH-C 1-6 alkyl, -NH-C 1-6 alkyl, C 1-6 alkyl-NH2, nitro, cyano and halogenated C 1-6 alkyl; and (11) R n1 and R n2 together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl group, which is optionally substituted with -NH-C 1-6 alkyl.

6. The compound of Formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-5, wherein R3 is selected from 7. The compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-6, wherein R4and R5together with the nitrogen atom to which they are attached form a 5- to 6-membered heterocycloalkenyl group.

8. The compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-7, wherein -NR4R5is selected from the following structures: Preferably 9. The compound of formula (I) according to any one of claims 1-6, which is a compound of formula (I-1), (I-2) or (I-3): ###00006### (I-1) (I-2) (I-3) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. ​ wherein, R3is as defined in any one of claims 1-6.

10. A compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

11. An antibody drug conjugate comprising a small molecule drug selected from the compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-10, a linker, and an antibody.

12. A pharmaceutical composition comprising the compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-10, and one or more pharmaceutically acceptable carriers.

13. Use of the compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-10, or the antibody drug conjugate according to claim 11, or the pharmaceutical composition according to claim 12, in the manufacture of a medicament for the treatment and / or prevention of a tumor; preferably the tumor is breast cancer, lung cancer, or colorectal cancer.

14. A method for the treatment and / or prevention of a tumor, comprising administering to a subject in need thereof an effective amount of the compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1-10, or an effective amount of the antibody drug conjugate according to claim 11, or an effective amount of the pharmaceutical composition according to claim 12; preferably the tumor is breast cancer, lung cancer, or colorectal cancer.

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