Camptothecin derivative and use thereof

Abstract

The present invention relates to a camptothecin derivative and a use thereof. In particular, the present invention relates to a compound represented by general formula (I), a preparation method therefor, a pharmaceutical composition thereof, and a use of the compound represented by general formula (I) or the pharmaceutical composition thereof. The compound can be used as a drug for treating diseases related to abnormal cell proliferation. Each substituent in general formula (I) is the same as defined in the description.

Description

Camptothecin derivatives and uses thereof TECHNICAL FIELD

[0001] The present application relates to a kind of brand-new structure camptothecin derivatives and antibody-drug conjugates thereof.The specific application relates to a kind of new camptothecin compounds, its stereoisomer or pharmaceutically acceptable salt and pharmaceutical composition, antibody-drug conjugate and the pharmaceutical composition comprising the conjugate, and its application in the medical field. BACKGROUND

[0002] Camptothecin is a very important natural product, mainly exists in the skin and fruit of the plant Camptotheca acuminata. The compound was first extracted from Camptotheca acuminata in 1966, and in vitro antitumor activity studies showed that camptothecin had strong antitumor activity against human cervical cancer cells Hela and mouse leukemia cells L1210 and other cancer cells. Camptothecin can exhibit strong antitumor activity mainly because it can inhibit topoisomerase I (Topo I). Topoisomerase I is widely present in mammalian body, and can play an important role in DNA replication, transcription and recombination.

[0003] Camptothecin has broad-spectrum antitumor activity in vitro, but due to its poor water solubility, instability in vivo and other defects, it cannot be directly used in clinical application, and its structure needs to be modified. After several generations of scientists' unremitting efforts, the structural modification of camptothecin as a parent and the corresponding drug research have achieved many results. Irinotecan, topotecan, belotecan, 10-hydroxycamptothecin and other drugs have been clinically applied, and dozens of derivatives are being studied in clinical or preclinical research.

[0004] In recent years, the emergence of antibody conjugate technology combining antibodies with various antitumor effector molecules (such as cytotoxic drugs, radioactive drugs and immunotoxins) has expanded the application range of camptothecin drugs in clinical treatment. Antibody-drug conjugates (ADC) is one of them, which connects highly specific monoclonal antibodies with cytotoxic drug carriers through a linker, uses the specific recognition of antibodies to target sites to accurately deliver cytotoxic drugs to tumor cells, reduces the damage of drugs to normal tissues and organs, breaks through the main clinical obstacles of traditional chemotherapy, and provides a wider therapeutic window.

[0005] In order to develop high-activity camptothecin anticancer drugs with practical application value in clinic more quickly and better, and to apply suitable camptothecin derivatives as cytotoxic drugs to ADC, it is necessary to develop new camptothecin drugs. SUMMARY

[0006] The application designs a series of new camptothecin compounds with good antitumor activity.

[0007] The application provides a compound shown in general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), general formula (II-4), general formula (III-1), general formula (III-2), general formula (III-3), general formula (IV-1), general formula (IV-2), general formula (L-I), general formula (L-IV-1), general formula (L-IV-2), general formula (Ab-L-I), general formula (Ab-L-IV-1), general formula (Ab-L-IV-2), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which has one or more effects selected from the following group: (1) has in-vitro proliferation inhibition activity on tumor cells; (2) has targeted inhibition; (3) has plasma stability; (4) has in-vivo tumor inhibition effect; (5) has a bystander effect; (6) has anti-transporter transport capacity; (7) has in-vivo tumor targeting capacity; and (8) has good in-vivo safety.

[0008] The application relates to a compound shown in general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), general formula (II-4), general formula (III-1), general formula (III-2), general formula (III-3), general formula (IV-1), general formula (IV-2), general formula (L-I), general formula (L-IV-1), general formula (L-IV-2), general formula (Ab-L-I), general formula (Ab-L-IV-1), general formula (Ab-L-IV-2), a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

[0009] wherein,

[0010] R1, R2, R3, R4 and R8 are each independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuterated alkyl, C 1-6 halogenated alkyl, C 1-6 alkoxy, C 1-6 halogenated alkoxy, C 3-6 cycloalkyl, 3-6-membered heterocycloalkyl, C6-10 aryl or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0011] In some embodiments, R1, R2, R3, R4, and R8 are each independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0012] In some embodiments, R1, R2, R3, R4, and R8 are each independently hydrogen, deuterium, halogen, or C 1-3 alkyl;

[0013] In some embodiments, R1 is hydrogen or deuterium; R2 is C 1-3 alkyl, R3 is halogen, and R4 is hydrogen, deuterium, or halogen;

[0014] In some embodiments, R2 is methyl;

[0015] In some embodiments, R3 is fluoro or chloro;

[0016] In some embodiments, R4 is hydrogen or deuterium;

[0017] In some embodiments, R4 is fluoro or chloro;

[0018] In some embodiments, R8 is hydrogen, deuterium, fluoro, chloro, methyl, ethyl;

[0019] In some embodiments, R2 is methyl, R3 is fluoro or chloro;

[0020] In some embodiments, R3 is fluoro or chloro, R4 is hydrogen or deuterium;

[0021] In some embodiments, R1 is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R, preferably cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0022] In some embodiments, R2is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R, preferably cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0023] In some embodiments, R1and R2, R3and R4, and the carbon atoms to which they are attached link to form a C 4-6 cycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0024] In some embodiments, R2and R3, and the carbon atoms to which they are attached link to form a 5-6 membered oxygen-containing heterocycloalkyl, optionally further substituted with 1-4 R;

[0025] In some embodiments, R1and R2, and the carbon atoms to which they are attached link to form a C 4-6 cycloalkyl, phenyl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0026] In some embodiments, R3and R4, and the carbon atoms to which they are attached link to form a C 4-6 cycloalkyl, phenyl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0027] In some embodiments, R2and R3, and the carbon atoms to which they are attached link to form a 5-6 membered heterocycloalkyl containing 2 oxygen atoms, optionally further substituted with 1-4 R;

[0028] In some embodiments, R1and R2, and the carbon atoms to which they are attached link to form a C 4-6 cycloalkyl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0029] In some embodiments, R1and R2, and the carbon atoms to which they are attached link to form cyclobutyl, cyclopentyl, cyclohexyl, furanyl, pyranyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, pyrrolyl, pyridyl, pyrimidinyl, pyridonyl, pyrazinyl, pyridazinyl, optionally further substituted with 1-4 R;

[0030] R5is hydrogen, deuterium, halogen, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C

[0031] In some embodiments, R5 is hydrogen, deuterium, halogen, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein said C

[0032] In some embodiments, R5 is C 2-6 alkyl, C 2-6 alkenyl, C

[0033] In some embodiments, R5 is ethenyl, ethynyl;

[0034] In some embodiments, R5 is hydrogen, deuterium, halogen, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C

[0035] In some embodiments, R5 is hydrogen, deuterium, halogen, methyl, ethyl, ethenyl, ethynyl;

[0036] R6 is hydrogen or links with either of R1, R8 to form C 5-8 cycloalkyl, wherein said C 5-8 cycloalkyl, wherein said C

[0037] In some embodiments, R6 is hydrogen or links with either of R1, R8 to form cyclohexyl, wherein said cyclohexyl is optionally further substituted with 1-4 R;

[0038] R7 is hydrogen, deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C1-6 haloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted with 1-4 R;

[0039] In some embodiments, R7is hydrogen, deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl are optionally further substituted with 1-4 R;

[0040] In some embodiments, R7is hydrogen, deuterium, halogen, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl;

[0041] In some embodiments, R7is hydrogen, deuterium, halogen, methyl, ethyl, ethenyl, ethynyl;

[0042] In some embodiments, R7is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0043] In some embodiments, R7is deuterium, fluorine, chlorine, methyl, ethyl, ethenyl, ethynyl;

[0044] A is -O-, -NH-, -O-(CR a R b ) m1 -C(O)NR c -, -S-(CR a R b ) m1 -C(O)NR c -, -O-(CRa R b ) m1 -C(O)NR c -NR c -、-NR d -(CR a R b ) m1 -C(O)NR c -、-O-(CR a R b ) m1 -C(O)NR d -(CR a R b ) m2 -C(O)NR c -、

[0045] In some embodiments, A is -O-, -NH-, -O-(CR a R b ) m1 -C(O)NR c -、-S-(CR a R b ) m1 -C(O)NR c -、-O-(CR a R b ) m1 -C(O)NR c -NR c -、-NR d -(CR a R b ) m1 -C(O)NR c -、-O-(CR a R b ) m1 -C(O)NR d -(CR a R b ) m2 -C(O)NR c -、

[0046] In some embodiments, A is -O-, -NH-, -O-CR a R b -C(O)NR c -、-S-CR a R b -C(O)NR c -、-O-CR a R b -C(O)NRc -NR c -、-NR d -CR a R b -C(O)NR c -、-O-CR a R b -C(O)NR d -CR a R b -C(O)NR c -、

[0047] In some embodiments, A is -O-, -NH-, -O-CR b -C(O)NH-, d -CHR b -C(O)NH-, c -, d -CHR b -C(O)NH-,

[0048] In some embodiments, A is -O-, -NH-, -O-CR a R b -C(O)NR c -, a R b -C(O)NR c -, a R b -C(O)NR c -NR c -, d -CR a R b -C(O)NR c -, a R b -C(O)NR d -CR a R b -C(O)NR c -,

[0049] In some embodiments, A is -O-, -NH-, -O-CR b -C(O)NH-, d -CHRb -C(O)NH-, -O-CH2-C(O)NR c -, -O-CH2-C(O)NR d -CHR b -C(O)NH-,

[0050] In some embodiments, A is -O-, -NH-, -O-CH2-C(O)NH-, -O-CH2-C(O)NH-NH-,

[0051] E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-,

[0052] In some embodiments, E is -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-,

[0053] In some embodiments, E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-,

[0054] In some embodiments, E is -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-, -O-CH2-C(O)NR d -CHR b -C(O)-,

[0055] In some embodiments, E is

[0056] each R a and R b is independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C 1-6 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl, or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl, or 5-6 membered heteroaryl is optionally further substituted with 1-4 R;

[0057] In some embodiments, each R a and R b is independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl, or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl, or 5-6 membered heteroaryl is optionally further substituted with 1-4 R;

[0058] In some embodiments, R a is hydrogen, deuterium, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C1-6 Alkylaminoalkyl, and R b C replaced by 1-4 Rs 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl;

[0059] In some implementations, R a For hydrogen, deuterium, C 1-3 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-3 Deuterated alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, C 1-3 Alkoxyalkyl, C 1-3 alkylamine group, C 1-3 Alkylaminoalkyl, and R b Cyclopropyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups;

[0060] In some implementations, R a It is hydrogen, and R b Cyclopropyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups;

[0061] In some implementations, R a For hydrogen, deuterium, halogens, C 1-3 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group;

[0062] In some implementations, R a It can be hydrogen, deuterium, halogen, methyl, ethyl, vinyl, or acetylene.

[0063] In some implementations, R a The compounds are deuterium, fluorine, chlorine, methyl, ethyl, vinyl, and ethynyl.

[0064] In some implementations, R b C replaced by 1-4 Rs 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl;

[0065] In some implementations, R b Cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups;

[0066] In some implementations, R b Cyclopropyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups;

[0067] In some implementations, R a and R b Each is independently hydrogen;

[0068] Rc and R d each independently is hydrogen, deuterium, C 1-6 alkyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, wherein the C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 R;

[0069] In some embodiments, R c and R d each independently is hydrogen, deuterium, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, wherein the C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 R;

[0070] In some embodiments, R d is hydrogen, deuterium, C 1-6 alkyl, preferably R d is hydrogen, deuterium, C 1-3 alkyl;

[0071] In some embodiments, R c and R d each independently is hydrogen;

[0072] Alternatively, in some embodiments, any two R a , any two R b , or any R a and R b link with the carbon atom to which they are attached to form a C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0073] Alternatively, in some embodiments, any R a and R b link with the carbon atom to which they are attached to form a C 3-6 cycloalkyl, optionally further substituted with 1-4 R;

[0074] Alternatively, in some embodiments, one of R a or R b and R dthe atom to which it is attached forms a 3-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R;

[0075] or, in some embodiments, R a or R b and R d the atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, and R is not halo, hydroxyl, oxo;

[0076] or, in some embodiments, R6and R c the atom to which it is attached forms a 3-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R;

[0077] or, in some embodiments, R8and R c the atom to which it is attached forms a 3-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R;

[0078] or, in some embodiments, R b and R d the atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, wherein R is not halo, hydroxyl, oxo;

[0079] or, in some embodiments, R6and R c the atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R;

[0080] or, in some embodiments, R8and R c the atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R;

[0081] in some embodiments, R b and R d the atom to which it is attached forms an azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl group, optionally further substituted with 1-4 R, wherein R is not halo, hydroxyl, oxo;

[0082] R is deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C

[0083] In some embodiments, R is deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C

[0084] In some embodiments, R is deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C1-3 haloalkyl, C 1-3 alkylalkylene, C 1-3 haloalkylalkylene, C 3-6 cycloalkylalkylene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkylalkylene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylene, or C 1-3 haloalkylalkylene, C

[0085] In some embodiments, R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylene, C 1-3 haloalkylalkylene, C 3-6 cycloalkylalkylene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylene, or C 1-3 haloalkylalkylene, C

[0086] In some embodiments, R is deuterium;

[0087] In some embodiments, R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy,

[0088] In some embodiments, R is methylene, ethylene, 1-methylethylene, fluoromethylene, difluoromethylene, cyclobutylmethylene, cyclopentylmethylene, cyclohexylmethylene;

[0089] R x1 is hydrogen, deuterium, halogen, amino, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylidene, C 1-6 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene;

[0090] In some embodiments, R x1 is C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene;

[0091] In some embodiments, R x1cycloalkyl, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene;

[0092] R x2 deutero, halo, amino, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylidene, C 1-6 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deutero, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene;

[0093] In some embodiments, R x2 is deutero;

[0094] n is 1, 2, 3, 4; in some embodiments, n is 1; in some embodiments, n is 2; in some embodiments, n is 3; in some embodiments, n is 4;

[0095] n1 is 1 or 2; n2 is 1 or 2;

[0096] m1 is 1, 2, 3; in some embodiments, m1 is 1, 2;

[0097] m2 is 1, 2, 3; in some embodiments, m2 is 1, 2;

[0098] n3 is 1, 2, 3, in some embodiments, n3 is 3; n4 is 1, 2, 3, in some embodiments, n4 is 1;

[0099] In some embodiments, L is L a -L2-L3-L4-, wherein:

[0100] L a is (maleimide-N-yl)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl), or -C 1-6 heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl containing 1-3 atoms selected from N, O, or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0101] L2 is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, wherein p is an integer from 0 to 20;

[0102] L3 is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0103] L4 is a bond, -NR L4 (CR L5 R L6 ) q -, -C(O)NR L4 (CH2) q -, wherein q is an integer from 0 to 6;

[0104] R L2 and R L4 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0105] R L5 and R L6 are each independently hydrogen, deuterium, halogen, C1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl;

[0106] In other embodiments, L is a linker unit, further -L1-L2-L3-L4-, L1 is attached to Ab and L4 is attached to A, wherein:

[0107] L1 is -(succinimid-3-yl-N)-Y-C(O)-, -CH2-C(O)-NR L1 -Y-C(O)-, -C(O)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl) or -C 1-6 heteroalkyl(3-6 membered heterocycloalkyl); said heteroalkyl contains 1-3 atoms selected from N, O or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl are optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C

[0108] L2 is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, wherein p is an integer from 0-20;

[0109] L3 is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C

[0110] L4 is a bond, -NR L4 (CR L5 R L6 ) q -, -C(O)NRL4 (CH2) q wherein q is an integer from 0-6;

[0111] R L2 and R L4 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0112] R L5 and R L6 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl;

[0113] Ab is an antibody, antibody fragment, or fragment linked to an antigen;

[0114] z is an integer or decimal number from 1-10;

[0115] In some embodiments, the L is the following structure: wherein p is an integer from 1-10;

[0116] In some embodiments, the L is the following structure: wherein p is an integer from 1-10, wherein the * end is attached to Ab;

[0117] In some embodiments, the L is the following structure:

[0118] In some embodiments, the L is the following structure: wherein the * end is attached to Ab;

[0119] z is an integer or decimal from 1 to 10; p is an integer from 0 to 20; in some embodiments, p is an integer from 0 to 6; q is an integer from 0 to 6; in some embodiments, p is 1, 2, 3, 4, 5, or 6; in some embodiments, q is 1, 2, 3, 4, or 5; in some embodiments, z is an integer or decimal from 2 to 8; in some embodiments, z is an integer or decimal from 3 to 8; in some embodiments, z is an integer or decimal from 3, 4, 5, 6, 7, or 8.

[0120] In some embodiments, the compounds of general formula (II-1), general formula (II-2), their stereoisomers, or pharmaceutically acceptable salts thereof satisfy at least one of the following conditions:

[0121] (1)R a For hydrogen, deuterium, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkoxyalkyl, C 1-6 alkylamine group, C 1-6 Alkylaminoalkyl, and R b C replaced by 1-4 Rs 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl,

[0122] In some implementations, R a For hydrogen, deuterium, C 1-3 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 1-3 Deuterated alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, C 1-3 Alkoxyalkyl, C 1-3 alkylamine group, C 1-3 Alkylaminoalkyl, and R b Cyclopropyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups;

[0123] (2) R2 is C 3-6 Cycloalkyl or 3-6 membered heterocyclic alkyl groups, optionally further substituted with 1-4 R groups.

[0124] In some embodiments, R2 is cyclopropyl, cyclobutyl, cyclopentyl, aziridine, oxacyclobutyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted by 1-4 Rs;

[0125] (3) R5 is C 2-6 alkenyl, C 2-6alkynyl,

[0126] In some embodiments, R5is vinyl, ethynyl;

[0127] (4) R7is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, preferably deuterium, fluorine, chlorine, methyl, ethyl, vinyl, ethynyl;

[0128] (5) R x1 is C 1-3 alkylalkylenyl, C 1-3 haloalkylalkylenyl, C 3-6 cycloalkylalkylenyl, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylenyl, or C 1-3 haloalkylalkylenyl,

[0129] In some embodiments, R x1 is cyclopropyl, cyclobutyl, cyclopentyl, optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylenyl, or C 1-3 haloalkylalkylenyl;

[0130] In some embodiments, the compound of general formula (II-3), general formula (II-4), stereoisomer thereof, or pharmaceutically acceptable salt thereof, satisfies at least one of the following conditions:

[0131] (1) R a or R b and R dwith the attached atoms forming a 4-6 membered heterocycloalkyl, optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo, or

[0132] (2) R2is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R,

[0133] In some embodiments, R2is cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0134] (3) R5is C 2-6 alkenyl, C 2-6 alkynyl,

[0135] In some embodiments, R5is ethenyl, ethynyl;

[0136] (4) R7is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 halogenated alkyl, C 1-3 alkoxy, C 1-3 halogenated alkoxy, preferably deuterium, fluorine, chlorine, methyl, ethyl, ethenyl, ethynyl;

[0137] (5) R x1 is C 1-3 alkylidene, C 1-3 halogenated alkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 halogenated alkyl, C 1-3 alkoxy, C 1-3 halogenated alkoxy, C 1-3 alkylidene, or C 1-3 halogenated alkylidene,

[0138] In some embodiments, R x1 is cyclopropyl, cyclobutyl, cyclopentyl, optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 halogenated alkyl, C1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene, substituted with;

[0139] In some embodiments, the compound of Formula (III-1), Formula (III-2), Formula (III-3), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, satisfies at least one of the following conditions:

[0140] (1) R1is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R,

[0141] In some embodiments, R1is cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0142] (2) R1and R2, together with the carbon atom to which they are attached, link to form C 4-6 cycloalkyl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0143] (3) R4is halogen;

[0144] (4) R x2 is deuterium.

[0145] In some embodiments, the compound of Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, satisfies at least one of the following conditions:

[0146] wherein:

[0147] R1, R2, R3, R4, and R8are each independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl is optionally further substituted with 1-4 R;

[0148] or, R1and R2, R3and R4, together with the carbon atom to which they are attached, link to form C 4-6 cycloalkyl, C6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0149] or, R2and R3, together with the carbon atom to which they are attached, link to form a 5-6 membered oxygen-containing heterocycloalkyl, optionally further substituted with 1-4 R;

[0150] R5is hydrogen, deuterium, halogen, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0151] R6is hydrogen or, together with either of R1, R8, forms a C 5-8 cycloalkyl, wherein the C 5-8 cycloalkyl is optionally further substituted with 1-4 R;

[0152] R7is hydrogen, deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0153] A is -O-, -NH-, -O-(CR a R b ) m1 -C(O)NR c -, -S-(CR a R b ) m1 -C(O)NR c -, -O-(CR a R b) m1 -C(O)NR c -NR c -、-NR d -(CR a R b ) m1 -C(O)NR c -、-O-(CR a R b ) m1 -C(O)NR d -(CR a R b ) m2 -C(O)NR c -、

[0154] In some embodiments, A is -O-, -NH-, -O-(CR a R b ) m1 -C(O)NR c -、-S-(CR a R b ) m1 -C(O)NR c -、-O-(CR a R b ) m1 -C(O)NR c -NR c -、-NR d -(CR a R b ) m1 -C(O)NR c -、-O-(CR a R b ) m1 -C(O)NR d -(CR a R b ) m2 -C(O)NR c -、

[0155] each R a and R b is independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6alkylamino, C 1-6 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0156] R c and R d are each independently hydrogen, deuterium, C 1-6 alkyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0157] or, any two R a , any two R b , or any R a and R b and the carbon atom to which they are attached link to form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0158] or, one of R a or R b and R d and the atoms to which they are attached link to form a 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0159] or, R6and R c and the atoms to which they are attached link to form a 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0160] or, R8and R c and the atoms to which they are attached link to form a 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0161] R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3haloalkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene; 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene;

[0162] n is 1, 2, 3, 4;

[0163] n1 is 1 or 2;

[0164] n2 is 1 or 2;

[0165] m1 is 1, 2, 3;

[0166] m2 is 1, 2, 3.

[0167] In particular, a second embodiment, the compound of Formula (I), stereoisomer thereof, or pharmaceutically acceptable salt thereof, satisfies one or more of the following conditions:

[0168] (1) R1, R2, R3, R4, and R8 are each independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl are optionally further substituted with 1-4 R;

[0169] or R1 and R2, together with the carbon atom to which they are attached, link to form a C 4-6 cycloalkyl, phenyl, or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0170] or R3 and R4, together with the carbon atom to which they are attached, link to form a C 4-6 cycloalkyl, phenyl, or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0171] or R2 and R3, together with the carbon atom to which they are attached, link to form a 5-6 membered heterocycloalkyl containing 2 oxygen atoms, optionally further substituted with 1-4 R;

[0172] (2) R5 is hydrogen, deuterium, halogen, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl are optionally further substituted with 1-4 R,

[0173] In some embodiments, R5 is hydrogen, deuterium, halogen, C 1-3 alkyl C 2-4 alkenyl, C 2-4 alkynyl,

[0174] In some embodiments, R5is hydrogen, deuterium, halogen, methyl, ethyl, ethenyl, ethynyl;

[0175] (3) R6is hydrogen or linked with either of R1, R8to form a cyclohexyl, wherein said cyclohexyl is optionally further substituted with 1-4 R;

[0176] (4) R7is hydrogen, deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl is optionally further substituted with 1-4 R,

[0177] In some embodiments, R7is hydrogen, deuterium, halogen, C 1-3 alkyl C 2-4 alkenyl, C 2-4 alkynyl,

[0178] In some embodiments, R7is hydrogen, deuterium, halogen, methyl, ethyl, ethenyl, ethynyl;

[0179] (4) A is -O-, -NH-, -O-CR a R b -C(O)NR c -, -S-CR a R b -C(O)NR c -, -O-CR a R b -C(O)NR c -NR c -, -NR d -CR a R b -C(O)NR c -, -O-CR a R b -C(O)NR d -CR a R b -C(O)NR c -,

[0180] In some embodiments, A is -0-, -NH-, -0-CR a R b -C(O)NR c -, a R b -C(O)NR c -, a R b -C(O)NR c -, c -, d -CR a R b -C(O)NR c -, a R b -C(O)NR d -CR a R b -C(O)NR c -,

[0181] In some embodiments, A is -0-, -NH-, -0-CH2-C(O)NH-, -0-CH2-C(O)NH-NH-, -O-CHR b -C(O)NH-, d -CHR b -C(O)NH-, c -0-CH2-C(O)NR d -CHR b -C(O)NH-,

[0182] In some embodiments, A is -0-, -NH-, -0-CH2-C(O)NH-, -0-CH2-C(O)NH-NH-, -O-CHR b -C(O)NH-, d -CHR b -C(O)NH-, c -0-CH2-C(O)NR d -CHR b -C(O)NH-,

[0183] (6) each R a and R b are each independently hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl or 5-6 membered heteroaryl are optionally further substituted with 1-4 R;

[0184] or, any R a and R b link with the carbon atom to which they are attached to form a C 3-6 cycloalkyl, optionally further substituted with 1-4 R;

[0185] (7) R c and R d are each independently hydrogen, deuterium, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl are optionally further substituted with 1-4 R,

[0186] In some embodiments, R c is hydrogen;

[0187] or, R b and R d link with the atom to which they are attached to form a 4-6 membered heterocycloalkyl, optionally further substituted with 1-4 R, wherein R is not halogen, hydroxyl, oxo;

[0188] or, R6and R c link with the atom to which they are attached to form a 4-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0189] or, R8and R c link with the atom to which they are attached to form a 4-6 membered heterocycloalkyl, optionally further substituted with 1-4 R;

[0190] (8) R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene,

[0191] In some embodiments, R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene,

[0192] In some embodiments, R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy,

[0193] (9) n is 1, 2, 3;

[0194] (10) m1and m2are each independently 1, 2.

[0195] In a particular third embodiment, the general formula (I) is further represented by general formula (II-1), general formula (II-2):

[0196] wherein R x1 is hydrogen, deuterium, halogen, amino, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylalkylene, C 1-6 haloalkylalkylene, C 3-6 cycloalkylalkylene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylene or C 1-3 haloalkylalkylene;

[0197] the remaining groups are defined as in any of the preceding embodiments.

[0198] In a particular fourth embodiment, the general formula (I) is further represented by general formula (II-3), general formula (II-4):

[0199] wherein R x1 is hydrogen, deuterium, halogen, amino, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylalkylene, C 1-6 haloalkylalkylene, C 3-6 cycloalkylalkylene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C3-6 cycloalkyl or 3-6 membered heterocycloalkyl optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene or C 1-3 haloalkylidene;

[0200] the remaining groups are as defined in any of the preceding embodiments.

[0201] In a particular fifth embodiment, the general formula (I) is further represented by general formula (III-1), (III-2), (III-3):

[0202] wherein R x2 is hydrogen, deuterium, halogen, amino, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylidene, C 1-6 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene or C 1-3 haloalkylidene;

[0203] n3 is 1, 2, 3, preferably 3; n4 is 1, 2, 3, preferably 1;

[0204] the remaining groups are as defined in any of the preceding embodiments.

[0205] In a particular sixth embodiment, the general formula (I) is further represented by general formula (IV-1), (IV-2):

[0206] wherein E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHRb -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, In some embodiments, E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, In some embodiments, E is -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-, -O-CH2-C(O)NR d -CHR b -C(O)-, In some embodiments, E is -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-, -O-CH2-C(O)NR d -CHR b -C(O)-,

[0207] R b is C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, in some embodiments, R b is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, substituted with 1-4 R, in some embodiments, R b is cyclopropyl, cyclopentyl, or cyclohexyl, substituted with 1-4 R;

[0208] R d is hydrogen, deuterium, C 1-6 alkyl, preferably R d is hydrogen, deuterium, C 1-3 alkyl;

[0209] or Rb and R d with the atom to which it is attached forming a 4-6 membered heterocycloalkyl, optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo, preferably R b and R d with the carbon atom to which it is attached forming an azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo;

[0210] R a , R, n1, n2, m1, and m2 are as described in any of the preceding embodiments;

[0211] In a particular seventh embodiment, the E is

[0212] In a particular eighth embodiment, the compound of general formula (II-1), general formula (II-2), stereoisomer thereof, or pharmaceutically acceptable salt thereof, satisfies at least one of the following conditions:

[0213] (1) R a is hydrogen, deuterium, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C 1-6 alkylaminoalkyl, and R b is cyclopropyl, cyclopentyl, or cyclohexyl substituted with 1-4 R; 3-6 cycloalkyl, 3-6 membered heterocycloalkyl,

[0214] In some embodiments, R a is hydrogen, deuterium, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, and R b is cyclopropyl, cyclopentyl, or cyclohexyl substituted with 1-4 R;

[0215] (2) R2is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R,

[0216] In some embodiments, R2is cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0217] (3) R5is C 2-6 alkenyl, C 2-6 alkynyl,

[0218] In some embodiments, R5is ethenyl, ethynyl;

[0219] (4) R7is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, preferably deuterium, fluorine, chlorine, methyl, ethyl, ethenyl, ethynyl;

[0220] (5) R x1 is C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene,

[0221] In some embodiments, R x1 is cyclopropyl, cyclobutyl, cyclopentyl, optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene.

[0222] In a ninth embodiment, the compounds of Formula (II-3), Formula (II-4), stereoisomers thereof, or pharmaceutically acceptable salts thereof, satisfy at least one of the following conditions:

[0223] (1) R a or R b and R d together with the atom to which they are attached form a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo; 3-6 2-6 2-6 1-3 2-4 2-4 1-3 1-3 1-3 1-3 x1 1-3 1-3 3-6 3-6 3-6 1-3 1-3 1-3 1-3 1-3 1-3 1-3 In a ninth embodiment, the compounds of Formula (II-3), Formula (II-4), stereoisomers thereof, or pharmaceutically acceptable salts thereof, satisfy at least one of the following conditions:

[0223] (1) R a or R b and R d together with the atom to which they are attached form a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo;

[0224] (2) R2is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R,

[0225] In some embodiments, R2is cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0226] (3) R5is C 2-6 alkenyl, C 2-6 alkynyl,

[0227] In some embodiments, R5is ethenyl, ethynyl;

[0228] (4) R7is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, preferably deuterium, fluorine, chlorine, methyl, ethyl, ethenyl, ethynyl;

[0229] (5) R x1 is C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene,

[0230] In some embodiments, R1is cyclopropyl, cyclobutyl, cyclopentyl, optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylalkylidene, or C 1-3 haloalkylalkylidene.

[0231] In a specific tenth embodiment, the compound of Formula (III-1), Formula (III-2), Formula (III-3), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, satisfies at least one of the following conditions:

[0232] (1) R1is C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, optionally further substituted with 1-4 R,

[0233] In some embodiments, R1is cyclopropyl, cyclobutyl, cyclopentyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted with 1-4 R;

[0234] (2) R1and R2are linked with the carbon atom to which they are attached to form C 4-6 cycloalkyl or 5-6 membered heteroaryl, optionally further substituted with 1-4 R;

[0235] (3) R4is halogen;

[0236] (4) R x2 is deuterium.

[0237] In a specific eleventh embodiment, the compound of Formula (I), Formula (II-1), Formula (II-2), Formula (II-3), Formula (II-4), Formula (III-1), Formula (III-2), Formula (III-3), Formula (IV-1), Formula (IV-2), is selected from the following Table 1:

[0238] Table 1:

[0239] In a specific twelfth embodiment, the present application relates to a linker-drug conjugate of Formula (L-I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0240] wherein:

[0241] L is L a -L2-L3-L4-, wherein:

[0242] L a is (maleimide-N-yl)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl), or -C 1-6 heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl containing 1-3 atoms selected from N, O, or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0243] L2 is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, wherein p is an integer from 0 to 20;

[0244] L3 is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0245] L4 is a bond, -NR L4 (CR L5 R L6 ) q -, -C(O)NR L4 (CH2) q -, wherein q is an integer from 0 to 6;

[0246] R L2 and R L4 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0247] R L5 and R L6 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl;

[0248] R1, R2, R3, R4, R5, R6, R7, R8, and A are as described in the preceding technical solution one or two;

[0249] In some embodiments, A is -O-, -NH-, -O-CH2-C(O)NH-, -O-CH2-C(O)NH-NH-,

[0250] In particular thirteenth embodiment, the general formula (L-I) is further shown as general formula (L-IV-1), general formula (L-IV-2):

[0251] wherein E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, preferably -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-, -O-CH2-C(O)NR d -CHR b -C(O)-, more preferably

[0252] R b is C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, preferably R bcyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, each of which is substituted with 1-4 R;

[0253] R d is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, preferably R d is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl;

[0254] or R b and R d form, together with the carbon atom to which they are attached, a nitrogen heterocyclobutyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, each of which is optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo; b and R d form, together with the carbon atom to which they are attached, a nitrogen heterocyclobutyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, each of which is optionally further substituted with 1-4 R, and R is not halogen, hydroxyl, oxo;

[0255] R a is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, preferably R 2-6 alkenyl, preferably R 2-6 alkynyl, preferably R 1-6 deuteroalkyl, preferably R 1-6 haloalkyl, preferably R 1-6 alkoxy, preferably R 1-6 alkoxyalkyl, preferably R 1-6 alkylamino, preferably R 1-6 alkylaminoalkyl, preferably R 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, preferably R 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, preferably R 6-10 aryl or 5-6 membered heteroaryl, is optionally further substituted with 1-4 R, preferably R is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, preferably R 2-4 alkenyl, preferably R 2-4 alkynyl, preferably R 1-3 deuteroalkyl, preferably R 1-3 haloalkyl, preferably R 1-3 alkoxy, preferably R 1-3 alkoxyalkyl, preferably R 1-3 alkylamino, preferably R 1-3 alkylaminoalkyl, preferably R 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, preferably R 6-8 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, preferably R 6-8 aryl or 5-6 membered heteroaryl, is optionally further substituted with 1-4 R;

[0256] R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 halogenated alkyl, C 1-3 alkoxy, C 1-3 halogenated alkoxy, C 1-3 alkylidene, C 1-3 halogenated alkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 halogenated alkyl, C 1-3 alkoxy, C 1-3 halogenated alkoxy, C 1-3 alkylidene or C 1-3 halogenated alkylidene, preferably deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuterated alkyl, C 1-3 halogenated alkyl, C 1-3 alkoxy, C 1-3 halogenated alkoxy,

[0257] n1 is 1 or 2; n2 is 1 or 2; m1 is 1, 2, 3; m2 is 1, 2, 3;

[0258] L is L a -L2-L3-L4-, wherein:

[0259] L a is (maleimide-N-yl)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl) or -C 1-6heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl comprising 1-3 atoms selected from N, O, or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl optionally further substituted with 1-4 groups independently selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0260] L2is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, wherein p is an integer from 0 to 20;

[0261] L3is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups independently selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0262] L4is a bond, -NR L4 (CR L5 R L6 ) q -, -C(O)NR L4 (CH2) q -, wherein q is an integer from 0 to 6;

[0263] R L2 and R L4 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0264] R L5 and R L6 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl.

[0265] In a particular fourteenth embodiment, L in the general formula (L-I), the general formula (L-IV-1), the general formula (L-IV-2) is the following structure: wherein p is an integer from 1 to 10.

[0266] In a particular fifteenth embodiment, the linker-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof in the general formula (L-I), the general formula (L-IV-1), the general formula (L-IV-2) is selected from one of the structures in Table II.

[0267] Table II:

[0268] In a particular fifteenth embodiment, the linker-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof in the general formula (L-I), the general formula (L-IV-1), the general formula (L-IV-2) is selected from one of the structures in Table II. the structure after removing the hydrogen.

[0269] Further, wherein the ligand-drug conjugate comprises the compound in the general formula (II-1), the general formula (II-2), the general formula (II-3), the general formula (II-4), the general formula (III-1), the general formula (III-2), the general formula (III-3) as previously described the structure after removing the hydrogen on the left, or the structure after removing the hydrogen on the left.

[0270] Further, wherein the ligand-drug conjugate comprises the following structure:

[0271] wherein the definitions of the groups are as described in any of the preceding embodiments.

[0272] In a particular sixteenth embodiment, an antibody-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof is represented by the general formula (Ab-L-I):

[0273] L is a linker unit, preferably -L1-L2-L3-L4-, L1 is attached to Ab, and L4 is attached to A, wherein:

[0274] L1 is -(succinimid-3-yl-N)-Y-C(O)-, -CH2-C(O)-NR L1 -Y-C(O)-, -C(O)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl), or -C1-6 heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl containing 1-3 atoms selected from N, O, or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0275] L2is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, wherein p is an integer from 0 to 20;

[0276] L3is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0277] L4is a bond, -NR L4 (CR L5 R L6 ) q -, -C(O)NR L4 (CH2) q -, wherein q is an integer from 0 to 6;

[0278] R L2 and R L4 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0279] R L5 and R L6 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6cycloalkylalkyl, 3-6 membered heterocycloalkyl;

[0280] Ab is an antibody, antibody fragment, or fragment linked to an antigen;

[0281] z is an integer or decimal number from 1 to 10;

[0282] R1, R2, R3, R4, R5, R6, R7, R8, and A are as described in the preceding technical solution one or two;

[0283] In some embodiments, A is -O-, -NH-, -O-CH2-C(O)NH-, -O-CH2-C(O)NH-NH-,

[0284] In a seventeenth embodiment, the general formula (Ab-L-I) is further illustrated by general formula (Ab-L-IV-1), general formula (Ab-L-IV-2):

[0285] wherein E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, preferably -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-, -O-CH2-C(O)NR d -CHR b -C(O)-, more preferably

[0286] R b is C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, preferably R b is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl substituted with 1-4 R;

[0287] R d is hydrogen, deuterium, C 1-6alkyl, preferably R d is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C

[0288] or R b and R d form together with the atom to which they are attached a 4-6 membered heterocycloalkyl, optionally further substituted by 1-4 R, and R is not halogen, hydroxyl, oxo, preferably R b and R d form together with the carbon atom to which they are attached an azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, optionally further substituted by 1-4 R, and R is not halogen, hydroxyl, oxo;

[0289] R a is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C 1-6 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted by 1-4 R, preferably R is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl or 5-6 membered heteroaryl are optionally further substituted by 1-4 R;

[0290] R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene or C 1-3 haloalkylidene, preferably deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy,

[0291] n1 is 1 or 2; n2 is 1 or 2; m1 is 1, 2, 3; m2 is 1, 2, 3;

[0292] L is a linker unit, preferably -L1-L2-L3-L4-, L1 is attached to Ab and L4 is attached to E, wherein:

[0293] L1 is -(succinimid-3-yl-N)-Y-C(O)-, -CH2-C(O)-NR L1 -Y-C(O)-, -C(O)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl) or -C 1-6heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl comprising 1-3 atoms selected from N, O, or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl optionally further substituted with 1-4 groups independently selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0294] L2is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, wherein p is an integer from 0 to 20;

[0295] L3is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups independently selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0296] L4is a bond, -NR L4 (CR L5 R L6 ) q -, -C(O)NR L4 (CH2) q -, wherein q is an integer from 0 to 6;

[0297] R L2 and R L4 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy;

[0298] R L5 and R L6 are each independently hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl;

[0299] Ab is an antibody, antibody fragment, or fragment linked to an antigen;

[0300] z is an integer or a decimal number from 1 to 10.

[0301] In a particular eighteenth embodiment, L in the general formula (Ab-L-I), general formula (Ab-L-IV-1), general formula (Ab-L-IV-2) is the following structure: wherein p is an integer from 1 to 10, and wherein the * end is attached to Ab.

[0302] In a particular nineteenth embodiment, the antibody-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof of the general formula (Ab-L-I), general formula (Ab-L-IV-1), general formula (Ab-L-IV-2) is selected from one of the structures in Table III, wherein Ab is an antibody, antibody fragment, or fragment linked to an antigen, and z is an integer or a decimal number from 1 to 10.

[0303] Table III:

[0304] In a particular twentieth embodiment, the antibody-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof of the general formula (Ab-L-I), general formula (Ab-L-IV-1), general formula (Ab-L-IV-2), wherein z is an integer or a decimal number from 2 to 8; in some embodiments, z is an integer or a decimal number from 3 to 8; in some embodiments, z is an integer or a decimal number from 3, 4, 5, 6, 7, 8.

[0305] In a twenty-first embodiment, the antibody-drug conjugate of Formula (Ab-L-I), Formula (Ab-L-IV-1), or Formula (Ab-L-IV-2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the Ab is selected from the group consisting of a murine antibody, a chimeric antibody, a humanized antibody, a fully human antibody, an antibody fragment, a bispecific antibody, and a multispecific antibody, preferably: an anti-EGFRvIII antibody, an anti-DLL-3 antibody, an anti-PSMA antibody, an anti-CD70 antibody, an anti-MUC16 antibody, an anti-ENPP3 antibody, an anti-TDGF1 antibody, an anti-ETBR antibody, an anti-MSLN antibody, an anti-TIM-1 antibody, an anti-LRRC15 antibody, an anti-LIV-1 antibody, an anti-CanAg / AFP antibody, an anti-cladin 18.2 antibody, an anti-Mesothelin antibody, an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-c-MET antibody, an anti-SLITRK6 antibody, an anti-KIT / CD117 antibody, an anti-STEAP1 antibody, an anti-SLAMF7 / CS1 antibody, an anti-NaPi2B / SLC34A2 antibody, an anti-GPNMB antibody, an anti-HER3 (ErbB3) antibody, an anti-MUC1 / CD227 antibody, an anti-AXL antibody, an anti-CD166 antibody, an anti-B7-H3 (CD276) antibody, an anti-PTK7 / CCK4 antibody, an anti-PRLR antibody, an anti-EFNA4 antibody, an anti-5T4 antibody, an anti-NOTCH3 antibody, an anti-Nectin 4 antibody, an anti-TROP-2 antibody, an anti-CD142 antibody, an anti-CA6 antibody, an anti-GPR20 antibody, an anti-CD174 antibody, an anti-CD71 antibody, an anti-EphA2 antibody, an anti-LYPD3 antibody, an anti-FGFR2 antibody, an anti-FGFR3 antibody, an anti-FRα antibody, an anti-CEACAMs antibody, an anti-GCC antibody, an anti-Integrin Av antibody, an anti-CAIX antibody, an anti-P-cadherin antibody, an anti-GD3 antibody, an anti-Cadherin 6 antibody, an anti-LAMP1 antibody, an anti-FLT3 antibody, an anti-BCMA antibody, an anti-CD79b antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CD56 antibody, an anti-CD74 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD37 antibody, an anti-CD47 antibody, an anti-CD138 antibody, an anti-CD352 antibody, an anti-CD25 antibody, an anti-CEACAM5 antibody, or an anti-CD123 antibody.

[0306] In some embodiments, the antibody-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof of general formula (Ab-L-I), general formula (Ab-L-IV-1), general formula (Ab-L-IV-2), the Ab is selected from: Trastuzumab, Pertuzumab, Nimotuzumab, Enoblituzumab, Emibetuzumab, Inotuzumab, Pinatuzumab, Brentuximab, Gemtuzumab, Bivatuzumab, Lorvotuzumab, Raludotatug, Sacituzumab, Labetuzumab, cBR96, and Glematumamab.

[0307] Secondly, the present application also provides a pharmaceutical composition containing the compound, ligand-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to any one of the preceding technical solutions, and one or more pharmaceutically acceptable carriers or excipients.

[0308] Further, the pharmaceutical composition or pharmaceutical preparation contains 1-1500 mg of the compound, ligand-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to any one of the preceding technical solutions, and one or more pharmaceutically acceptable carriers or excipients.

[0309] Further, the present application also provides the use of the compound, ligand-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of the preceding embodiments in the preparation of a medicament, preferably a medicament for preventing and / or treating a cell abnormal proliferation disease; in some embodiments, the cell abnormal proliferation disease is a tumor; in some embodiments, the cell abnormal proliferation disease is a solid tumor and a blood tumor.

[0310] The present application also provides a method for treating a disease in a mammal, the method comprising administering to the subject a therapeutically effective amount of the compound, ligand-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to any one of the preceding technical solutions, and a pharmaceutically acceptable carrier and / or excipient, the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably a tumor.

[0311] In some embodiments, the tumor is selected from the group consisting of breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, glioblastoma multiforme, lymphoma, lung cancer, colon cancer, rectal cancer, colorectal cancer, leukemia, bone cancer, skin cancer, thyroid cancer, pancreatic cancer.

[0312] The present application also provides a method for treating a disease in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound, a ligand drug conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present application. In some embodiments, the mammal of the present application includes a human.

[0313] An "effective amount" or "therapeutically effective amount" as described herein refers to an amount of a compound disclosed herein that, when administered, will relieve to some extent one or more of the symptoms of the disease or condition being treated. In some embodiments, the result is a decrease and / or alleviation of symptoms, signs, or causes of a disease, disorder, or condition, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic uses is the amount of a compound, conjugate, or pharmaceutically acceptable salt thereof disclosed herein that will elicit the biological or medical response (e.g., decrease or alleviation of a disease symptom, signs, or causes) in a tissue system, animal, or human. Examples of therapeutic effects include, but are not limited to, 1-1500 mg, 1-1400 mg, 1-1300 mg, 1-1200 mg, 1-1000 mg, 1-900 mg, 1-800 mg, 1-700 mg, 1-600 mg, 1-500 mg, 1-400 mg, 1-300 mg, 1-250 mg, 1-200 mg, 1-150 mg, 1-125 mg, 1-100 mg, 1-80 mg, 1-60 mg, 1-50 mg, 1-40 mg, 1-25 mg, 1-20 mg, 5-1500 mg, 5-1000 mg, 5-900 mg, 5-800 mg, 5-700 mg, 5-600 mg, 5-500 mg, 5-400 mg, 5-300 mg, 5-250 mg, 5-200 mg, 5-150 mg, 5-125 mg, 5-100 mg, 5-90 mg, 5-70 mg, 5-80 mg, 5-60 mg, 5-50 mg, 5-40 mg, 5-30 mg, 5-25 mg, 5-20 mg, 10-1500 mg, 10-1000 mg, 10-900 mg, 10-800 mg, 10-700 mg, 10-600 mg, 10-500 mg, 10-450 mg, 10-400 mg, 10-300 mg, 10-250 mg, 10-200 mg, 10-150 mg, 10-125 mg, 10-100 mg, 10-90 mg, 10-80 mg, 10-70 mg, 10-60 mg, 10-50 mg, 10-40 mg, 10-30 mg, 10-20 mg; 20-1500 mg, 20-1000 mg, 20-900 mg, 20-800 mg, 20-700 mg, 20-600 mg, 20-500 mg, 20-400 mg, 20-350 mg, 20-300 mg, 20-250 mg, 20-200 mg, 20-150 mg, 20-125 mg, 20-100 mg, 20-90 mg, 20-80 mg, 20-70 mg, 20-60 mg, 20-50 mg, 20-40 mg, 20-30 mg;50-1500 mg, 50-1000 mg, 50-900 mg, 50-800 mg, 50-700 mg, 50-600 mg, 50-500 mg, 50-400 mg, 50-300 mg, 50-250 mg, 50-200 mg, 50-150 mg, 50-125 mg, 50-100 mg; 100-1500 mg, 100-1000 mg, 100-900 mg, 100-800 mg, 100-700 mg, 100-600 mg, 100-500 mg, 100-400 mg, 100-300 mg, 100-250 mg, 100-200 mg;

[0314] In some embodiments, the pharmaceutical composition or formulation of the present application contains a therapeutically effective amount of the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof as described in any of the above.

[0315] The present application further relates to a pharmaceutical composition or a pharmaceutical formulation comprising a therapeutically effective amount of the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof as described in any of the above and one or more pharmaceutically acceptable carriers or excipients. The pharmaceutical composition can be in the form of a unit formulation (the amount of the principal drug in the unit formulation is also referred to as "formulation strength"). In some embodiments, the pharmaceutical composition includes, but is not limited to, 1 mg, 1.25 mg, 2.5 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg of the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof as described in any of the above.

[0316] The present application further relates to a method for treating a disease in a mammal, the method comprising administering to the subject a compound, stereoisomer, or pharmaceutically acceptable salt thereof, of any one of the above described compounds of the present application, and one or more pharmaceutically acceptable carriers or excipients, in a daily dose of 1-1500 mg per day, which can be in a single dose or divided doses, in some embodiments, the daily dose includes, but is not limited to, 10-1500 mg per day, 20-1500 mg per day, 25-1500 mg per day, 50-1500 mg per day, 75-1500 mg per day, 100-1500 mg per day, 200-1500 mg per day, 10-1000 mg per day, 20-1000 mg per day, 25-1000 mg per day, 50-1000 mg per day, 75-1000 mg per day, 100-1000 mg per day, 200-1000 mg per day, 25-800 mg per day, 50-800 mg per day, 100-800 mg per day, 200-800 mg per day, 25-400 mg per day, 50-400 mg per day, 100-400 mg per day, 200-400 mg per day, in some embodiments, the daily dose includes, but is not limited to, 1 mg per day, 5 mg per day, 10 mg per day, 20 mg per day, 25 mg per day, 50 mg per day, 75 mg per day, 100 mg per day, 125 mg per day, 150 mg per day, 200 mg per day, 300 mg per day, 400 mg per day, 600 mg per day, 800 mg per day, 1000 mg per day, 1200 mg per day, 1400 mg per day, 1500 mg per day.

[0317] The present application relates to a kit, which can include a single dose or multiple dose forms of a composition, the kit comprising a compound, stereoisomer, or pharmaceutically acceptable salt thereof, of any one of the above described compounds of the present application, the amount of the compound of the present application or the stereoisomer or pharmaceutically acceptable salt thereof in the kit being the same as the amount thereof in the above described pharmaceutical composition.

[0318] The amount of the compound of the present application or the stereoisomer or pharmaceutically acceptable salt thereof in the present application is converted into the form of free base in each case.

[0319] "Formulation strength" refers to the weight of the principal drug contained in each unit of preparation.

[0320] Synthetic route

[0321] The compounds of the present application can be prepared by those skilled in the art of organic synthesis by adapting known synthetic procedures in conjunction with known starting materials which are either commercially available or described in the chemical literature. "Commercially available chemicals" are obtained from regular commercial sources, including suppliers such as Titan Kogyo, Acros Organics, Sigma-Aldrich, Shanghai Chem-Ex, Chengdu Kelong Chemical, Shaoyuan Chemical Technology, Nanjing Huashen, WuXi AppTec and Bailingwei Technology.

[0322] Specific and analogous reactants can be identified selectively by the use of the Index of Known Chemical Substances prepared by the Chemical Abstract Service of the American Chemical Society, which is available in most public and university libraries, and on-line. Chemicals that are known but not commercially available can alternatively be prepared by custom chemical synthesis houses, many of which standard chemical supply houses (such as those listed above) offer custom synthesis services.

[0323] Definitions

[0324] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. If there is a conflict between the definitions provided in this application and those in dictionaries or other data sources, the definitions that are provided in this application control. Where a term is present in the singular, the plural is also contemplated, unless the context clearly dictates otherwise. All patents, published patent applications and publications mentioned herein are incorporated by reference.

[0325] The term "alkyl" refers to saturated straight chain or branched chain aliphatic hydrocarbon groups having from 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) carbon atoms, i.e., "C 1-20 The alkyl groups preferably have from 1 to 12 carbon atoms (i.e., C 1-12 The alkyl groups preferably have from 1 to 12 carbon atoms (i.e., C 1-8 The alkyl groups preferably have from 1 to 12 carbon atoms (i.e., C 1-6 The alkyl groups preferably have from 1 to 12 carbon atoms (i.e., C 1-3Non-limiting examples include: methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof, and the like. The alkyl group can be substituted or unsubstituted, and when substituted, the substituents can be substituted at any available attachment point. When the alkyl group is substituted with a substituent, the substituent is not further substituted.

[0326] The term "heteroalkyl" refers to saturated straight-chain or branched-chain aliphatic hydrocarbon groups consisting of a certain number of carbon atoms and at least one heteroatom. In one exemplary embodiment, the heteroatom is selected from B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. The heteroatoms B, O, N, and S can be located at any interior position of the heteroalkyl group, including the position at which the hydrocarbyl group is attached to the rest of the molecule, but not at the terminal position of the heteroalkyl group. The numerical ranges are for the number of carbon atoms in the chain, C 1-6Heteroalkyl refers to includes 1-6 carbon atoms, for example, containing 2, 3, 4, 5, or 6 carbon atoms. For example, a -CH2CH2OCH2CH3group is referred to as a C4heteroalkyl. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, -CH2-CH2-O-CH2-CH2-, -CH2-CH2-O-CH2-CH2-O-CH2-CH2-. Up to two heteroatoms can be consecutive, such as -CH2-NH-OCH3.

[0327] The term "alkylene" refers to a divalent linear and branched saturated alkyl group. Examples of alkylene include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), and the like.

[0328] The term "alkenyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon double bond (C=C), typically containing 2 to 18 carbon atoms, such as 2 to 8 carbon atoms, further such as 2 to 6 carbon atoms, yet further such as 2 to 4 carbon atoms, examples of which include, but are not limited to, ethenyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1,3-butadiene, 1,3-pentadiene, 1,4-pentadiene, and 1,4-hexadiene, and the like; the alkenyl group can be substituted or non-substituted, when substituted, the substituents can be substituted at any available attachment point. When the alkenyl group is substituted with a substituent, the substituent is not further substituted.

[0329] The term "alkynyl" refers to a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond (C≡C), typically containing 2 to 18 carbon atoms, further containing 2 to 8 carbon atoms, further containing 2 to 6 carbon atoms, still further containing 2 to 4 carbon atoms, examples of which include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonylnyl, and 4-decynyl, and the like; the alkynyl group can be substituted or non-substituted, and when substituted, the substituents can be substituted on any available attachment point. When the alkynyl group is substituted with a substituent, the substituent is not further substituted.

[0330] The term "heterocycle" or "heterocyclyl" refers to a substituted or unsubstituted, saturated or unsaturated aromatic or non-aromatic ring, containing, unless otherwise specified, 1 to 3 heteroatoms selected from N, O, or S, including monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic fused heterocycles, and bicyclic spiro heterocycles, and the like, and unless otherwise specified, 3 to 12 membered heterocycles, more preferably 4 to 12 membered heterocycles, more preferably 4 to 10 membered heterocycles, and further preferably 4 to 7 membered heterocycles. The definition includes heterocycloalkyl and heteroaryl groups. The N, S in the heterocyclyl ring can be oxidized to various oxidation states. The heterocyclyl group can be attached at a heteroatom or carbon atom, non-limiting examples include oxiranyl, aziridinyl, oxetanyl, azetidinyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxananyl, azepanyl, pyridyl, furanyl, thienyl, pyranyl, N-alkyl pyrrolyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, imidazolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl, dihydrofuranyl, dihydropyranyl, dihydrothienyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, oxazolyl, dihydrooxazolyl, tetrahydrooxazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, benzodihydrofuranyl, azabicyclo[3.2.1]octanyl, azabicyclo[5.2.0]nonanyl, oxatricyclo[5.3.1.1]dodecanyl, azadamantanyl, and oxaspiro[3.3]heptanyl, and the like.

[0331] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon substituent (i.e., monocyclic cycloalkyl) or a polycyclic cyclic hydrocarbon substituent (i.e., polycyclic cycloalkyl) having 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) carbon atoms, i.e., C 3-20 The cycloalkyl group preferably has a cycloalkyl group having 3 to 12 carbon atoms (i.e., C 3-12cycloalkyl), more preferably a cycloalkyl having from 3 to 8 carbon atoms (i.e., C 3-8 cycloalkyl), more preferably a cycloalkyl having from 3 to 8 carbon atoms (i.e., C 3-6 cycloalkyl), more preferably a cycloalkyl having from 3 to 8 carbon atoms (i.e., C 3-5 cycloalkyl). Non-limiting examples of monocyclic cycloalkyl groups include: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, and cyclooctyl, and the like. Non-limiting examples of polycyclic cycloalkyl groups include: spirocycloalkyl, fused cycloalkyl, and bridged cycloalkyl.

[0332] The term "spirocycloalkyl" refers to a polycyclic group sharing one carbon atom (termed a spiro atom) between single rings, which can contain one or more double bonds, but no ring has a fully conjugated pi-electron system, which has from 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e., C 5-20 spirocycloalkyl). The spirocycloalkyl group preferably has from 6 to 14 ring atoms (i.e., C 6-14 spirocycloalkyl), more preferably a spirocycloalkyl having from 7 to 10 ring atoms (i.e., C 7-10 spirocycloalkyl). The spirocycloalkyl group is classified as a mono-, bi-, or polycycloalkyl depending on the number of spiro atoms shared between rings, preferably a mono- or bi-cycloalkyl, more preferably a 3-membered / 4-membered, 3-membered / 5-membered, 3-membered / 6-membered, 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 3-membered, 5-membered / 4-membered, 5-membered / 5-membered, 5-membered / 6-membered, 5-membered / 7-membered, 6-membered / 3-membered, 6-membered / 4-membered, 6-membered / 5-membered, 6-membered / 6-membered, 6-membered / 7-membered, 7-membered / 5-membered, or 7-membered / 6-membered spirocycloalkyl.

[0333] The term "fused cycloalkyl" refers to an all-carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, which has from 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e., C 5-20 fused cycloalkyl). It can contain one or more double bonds, but no ring has a fully conjugated pi-electron system. The fused cycloalkyl group preferably has from 6 to 14 ring atoms (i.e., C 6-14 fused cycloalkyl), more preferably a fused cycloalkyl having from 7 to 10 ring atoms (i.e., C 7-10Fused ring alkyl). It is classified as bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl depending on the number of rings comprising it, preferably bicyclic fused ring alkyl or tricyclic fused ring alkyl, more preferably 3- membered / 4-membered, 3-membered / 5-membered, 3-membered / 6-membered, 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 3-membered, 5-membered / 4-membered, 5-membered / 5-membered, 5-membered / 6-membered, 5-membered / 7-membered, 6-membered / 3-membered, 6-membered / 4-membered, 6-membered / 5-membered, 6-membered / 6-membered, 6-membered / 7-membered, 7-membered / 5-membered or 7-membered / 6-membered bicyclic fused ring alkyl.

[0334] The term "bridged ring alkyl" refers to an all-carbon polycyclic group which shares two non-adjacent carbon atoms between any two rings, having 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e., C 5-20 Bridged ring alkyl). It contains one or more double bonds, but no ring has a fully conjugated pi-electron system. The bridged ring alkyl preferably has 6 to 14 ring atoms (i.e., C 6-14 Bridged ring alkyl), more preferably 7 to 10 ring atoms (i.e., C 7-10 Bridged ring alkyl). It is classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged ring alkyl depending on the number of rings comprising it, preferably bicyclic bridged ring alkyl or tricyclic bridged ring alkyl.

[0335] The cycloalkyl group includes polycyclic cycloalkyl groups which can be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the rings which are connected together with the parent structure are cycloalkyl groups, for example including C 5-6 Cycloalkyl and phenyl, C 5-6 Cycloalkyl and 5-6 membered heteroaryl, C 5-6 Cycloalkyl and 5-6 membered heterocycloalkyl, etc. The cycloalkyl group can be optionally substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment. When the cycloalkyl group is substituted with a substituent, the substituent is not further substituted.

[0336] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic heterocyclic hydrocarbon substituent (i.e., monocyclic heterocycloalkyl) or a polycyclic heterocyclic hydrocarbon substituent (i.e., polycyclic heterocycloalkyl) having 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e., 3-20 membered heterocycloalkyl), wherein one or more (e.g., 1, 2, 3, or 4) ring atoms are selected from nitrogen, oxygen, P(O) m and S(O) nheteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. The heterocycloalkyl group preferably has 3 to 12 ring atoms (i.e., 3-12 membered heterocycloalkyl), more preferably 3 to 8 ring atoms (i.e., 3-8 membered heterocycloalkyl), further preferably 3 to 6 ring atoms (i.e., 3-6 membered heterocycloalkyl), and most preferably 5 to 6 ring atoms (i.e., 5-6 membered heterocycloalkyl), wherein 1 to 4, 1 to 3, or 1 to 2 of the ring atoms are heteroatoms selected from N, O, and S. Non-limiting examples of monocyclic heterocycloalkyl groups include azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, piperidinyl, piperazinyl, morpholinyl, 1,3-dioxolanyl, 2,2-difluoro-1,3-dioxolanyl, cyclopentanonyl, 2,2-difluorocyclopentanonyl, azepinyl, oxolanyl, or azolidinyl, and the like. Non-limiting examples of polycyclic heterocycloalkyl groups include spiroheterocycloalkyl, fused heterocycloalkyl, and bridged heterocycloalkyl.

[0337] The term "spiroheterocycloalkyl" refers to a polycyclic heterocycloalkyl group that shares one atom (referred to as a spiro atom) between single rings, which has 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e., 5-20 membered spiroheterocycloalkyl), wherein one or more (e.g., 1, 2, 3, or 4) of the ring atoms are selected from nitrogen, oxygen, P(O) m and S(O) n heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. It can contain one or more double bonds, but no ring has a fully conjugated pi-electon system. The spiroheterocycloalkyl group preferably has 6 to 14 ring atoms (i.e., 6-14 membered spiroheterocycloalkyl), more preferably 7 to 10 ring atoms (i.e., 7-10 membered spiroheterocycloalkyl). The spiroheterocycloalkyl group is classified as a mono-, bi-, or polycyclic spiroheterocycloalkyl group depending on the number of spiro atoms shared between the rings, preferably a mono- or bi-spiroheterocycloalkyl group, more preferably a 3 / 4-, 3 / 5-, 3 / 6-, 4 / 4-, 4 / 5-, 4 / 6-, 5 / 3-, 5 / 4-, 5 / 5-, 5 / 6-, 5 / 7-, 6 / 3-, 6 / 4-, 6 / 5-, 6 / 6-, 6 / 7-, 7 / 5-, or 7 / 6 mono-spiroheterocycloalkyl group.

[0338] The term "fused heterocycloalkyl" or "annelated heterocycloalkyl" refers to a polycyclic heterocycloalkyl group in which each ring shares an adjacent pair of atoms with other rings in the system, having 5 to 20 (e.g. 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e. 5-20 membered fused heterocycloalkyl), wherein one or more (e.g. 1, 2, 3, or 4) ring atoms are selected from nitrogen, oxygen, P(O) m and S(O) n (herein m, n are integers from 0 to 2) but excluding ring members -0-0-, -0-S-, or -S-S-, the remaining ring atoms being carbon. It can contain one or more double bonds, but no ring has a completely conjugated pi-electron system. The fused heterocycloalkyl group is preferably a fused heterocycloalkyl group having 6 to 14 ring atoms (i.e. 6-14 membered fused heterocycloalkyl), more preferably a fused heterocycloalkyl group having 7 to 10 ring atoms (i.e. 7-10 membered fused heterocycloalkyl). It is classified as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocycloalkyl group according to the number of rings making up the fused heterocycloalkyl group, preferably a bicyclic fused heterocycloalkyl group or a tricyclic fused heterocycloalkyl group, more preferably a 3 / 4, 3 / 5, 3 / 6, 4 / 4, 4 / 5, 4 / 6, 5 / 3, 5 / 4, 5 / 5, 5 / 6, 5 / 7, 6 / 3, 6 / 4, 6 / 5, 6 / 6, 6 / 7, 7 / 5, or 7 / 6 bicyclic fused heterocycloalkyl group.

[0339] The term "bridged heterocycloalkyl" refers to a polycyclic heterocycloalkyl group in which any two rings share two non-adjacent atoms, having 5 to 20 (e.g. 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) ring atoms (i.e. 5-20 membered bridged heterocycloalkyl), wherein one or more (e.g. 1, 2, 3, or 4) ring atoms are selected from nitrogen, oxygen, P(O) m and S(O) n (herein m, n are integers from 0 to 2) but excluding ring members -0-0-, -0-S-, or -S-S-, the remaining ring atoms being carbon. It can contain one or more double bonds, but no ring has a completely conjugated pi-electron system. The bridged heterocycloalkyl group is preferably a bridged heterocycloalkyl group having 6 to 14 ring atoms (i.e. 6-14 membered bridged heterocycloalkyl), more preferably a bridged heterocycloalkyl group having 7 to 10 ring atoms (i.e. 7-10 membered bridged heterocycloalkyl). It is classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocycloalkyl group according to the number of rings making up the bridged heterocycloalkyl group, preferably a bicyclic bridged heterocycloalkyl group or a tricyclic bridged heterocycloalkyl group.

[0340] The heterocycloalkyl groups include polycyclic heterocycloalkyl groups that can be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring that is attached to the parent structure is a heterocycloalkyl ring, including, for example, 5-6 membered heterocycloalkyl and phenyl, 5-6 membered heterocycloalkyl and 5-6 membered heteroaryl, 5-6 membered heterocycloalkyl and C 5-6 cycloalkyl, etc., preferably 5 membered heterocycloalkyl and 5 membered heterocycloalkyl, 5 membered heterocycloalkyl and 6 membered heterocycloalkyl, 5 membered heterocycloalkyl and 5 membered heteroaryl, 5 membered heterocycloalkyl and 6 membered heteroaryl, 6 membered heterocycloalkyl and 6 membered heterocycloalkyl, 6 membered heterocycloalkyl and 5 membered heteroaryl, 6 membered heterocycloalkyl and 6 membered heteroaryl, etc. The heterocycloalkyl groups can be optionally substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment. When the heterocycloalkyl groups are substituted with substituents, the substituents are not further substituted.

[0341] The term "aryl" refers to all-carbon monocyclic (i.e., single ring aryl) or fused polycyclic (i.e., multiple ring aryl) groups having a conjugated pi-electron system, having 6 to 14 (e.g., 6, 7, 8, 9, 10, 11, 12, 13, or 14) carbon atoms (i.e., C 6-14 aryl). The aryl groups preferably have 6 to 12 carbon atoms (i.e., C 6-12 aryl), more preferably 6 to 10 carbon atoms (i.e., C 6-10 aryl), even more preferably phenyl or naphthyl, most preferably phenyl. The monocyclic aryl groups, such as phenyl. Non-limiting examples of polycyclic aryl groups include: naphthyl, anthryl, phenanthryl, etc.

[0342] The aryl groups include polycyclic systems that can be fused to a heterocycloalkyl or cycloalkyl ring, wherein the ring that is attached to the parent structure is an aryl ring, including, but not limited to, benzo C 3-8 cycloalkyl, benzo 3-8 membered heterocycloalkyl, preferably benzo C 4-6 cycloalkyl, benzo 4-6 membered heterocycloalkyl, even more preferably benzo cyclobutyl, benzo cyclopentyl, benzo cyclohexyl, benzo azetidinyl, benzo oxetanyl, benzo oxetanyl, benzo oxetanyl, benzo oxetanyl, benzo oxetanyl, benzo oxetanyl, etc. The aryl groups can be optionally substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment. When the aryl groups are substituted with substituents, the substituents are not further substituted.

[0343] The term "heteroaryl" refers to monocyclic heteroaryl groups (i.e., single ring heteroaryl) or fused polycyclic heteroaryl groups (i.e., multiple ring heteroaryl) having a conjugated pi-electron system, having 5 to 14 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) ring atoms (i.e., 5-14 membered heteroaryl), wherein one or more (e.g., 1, 2, 3, or 4) ring atoms are selected from nitrogen, oxygen, P(O)m and S(O) n wherein m, n are integers from 0 to 2, and the heteroatom is preferably a heteroatom selected from nitrogen, oxygen, or sulfur, but excluding -O-O-, -O-S-, or -S-S- ring members, and the remaining ring atoms are carbon. The heteroaryl group preferably has from 5 to 10 ring atoms (i.e., 5-10 membered heteroaryl). The monocyclic heteroaryl group preferably has from 5 to 6 ring atoms (i.e., 5-6 membered heteroaryl), non-limiting examples of which include furyl, pyranyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, pyrrolyl, pyridyl, pyrimidinyl, pyridonyl, pyrazinyl, pyridazinyl, and the like. The polycyclic heteroaryl group preferably has from 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-10 membered heteroaryl and C 6-10 aryl or C 6-10 aryl and 5-10 membered heteroaryl, further preferably 5-6 membered heteroaryl and 5-6 membered heteroaryl, 5-6 membered heteroaryl and phenyl, or phenyl and 5-6 membered heteroaryl, non-limiting examples of which include indolyl, indazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, phthalazinyl, benzimidazolyl, benzothienyl, thienophenyl, quinazolinyl, benzothiazolyl, carbazolyl, thienopyridyl, pyridothienyl, pyridopyrrolyl, benzo-γ-pyrone, pyrido-γ-pyrone, and the like.

[0344] The heteroaryl group includes polycyclic ring systems fused to an aryl, heterocycloalkyl, or cycloalkyl ring, wherein the ring that is attached to the parent structure is a heteroaryl ring, including but not limited to 5-6 membered heteroaryl and C 3-8 cycloalkyl, 5-6 membered heteroaryl and 3-8 membered heterocycloalkyl, 5-6 membered heteroaryl and phenyl, preferably 5-6 membered heteroaryl and C 4-6 cycloalkyl, 5-6 membered heteroaryl and 4-6 membered heterocycloalkyl, 5-6 membered heteroaryl and phenyl. The heteroaryl group can be optionally substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment. When the heteroaryl group is substituted with a substituent, the substituent is not further substituted. Non-limiting examples of which include: and the like.

[0345] The term "alkoxy" refers to -O-(alkyl) or -O-(unsubstituted cycloalkyl), wherein alkyl, cycloalkyl are as defined above, having from 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) carbon atoms (i.e., C 1-10 alkoxy). The alkoxy group preferably has from 1 to 8 carbon atoms (i.e., C 1-8 alkoxy), more preferably from 1 to 6 carbon atoms (i.e., C 1-6alkyl) or -S-(unsubstituted cycloalkyl), wherein alkyl, cycloalkyl are defined as above, having 1 to 10 (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon atoms (i.e. C 1-3 alkoxy). Non-limiting examples include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexyloxy and the like. The alkoxy group can be optionally substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment. When the alkoxy group is substituted with a substituent, the substituent is not further substituted.

[0346] The term "alkylthio" means -S-(alkyl) or -S-(unsubstituted cycloalkyl), wherein alkyl, cycloalkyl are defined as above, having 1 to 10 (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon atoms (i.e. C 1-10 alkylthio). The alkylthio group preferably has 1 to 8 carbon atoms (i.e. C 1-8 alkylthio). The alkylthio group more preferably has 1 to 6 carbon atoms (i.e. C 1-6 alkylthio). The alkylthio group most preferably has 1 to 3 carbon atoms (i.e. C 1-3 alkylthio). Non-limiting examples include: methylthio, ethylthio, propylthio, butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio and the like. The alkylthio group can be optionally substituted or unsubstituted, and when substituted, the substituents can be substituted at any available point of attachment. When the alkylthio group is substituted with a substituent, the substituent is not further substituted.

[0347] The term "halo" or "halogen" or "halogenated" is understood to mean a fluorine (F), chlorine (CI), bromine (Br) or iodine (I) atom, preferably a fluorine, chlorine, bromine atom.

[0348] The term "haloalkyl" refers to an alkyl group, as defined above, substituted with one or more halogen. Non-limiting examples include fluoromethyl, chloromethyl, bromomethyl, iodomethyl, difluoromethyl, chlorofluoromethyl, dichloromethyl, bromofluoromethyl, trifluoromethyl, chlorodifluoromethyl, dichlorofluoromethyl, trichloromethyl, bromodifluoromethyl, bromochlorofluoromethyl, dibromofluoromethyl, and the like; preferably fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 2,2-difluoroethyl, 2-chloro-2-fluoroethyl, 2,2-dichloroethyl, 2-bromo-2-fluoroethyl, 2,2,2-trifluoroethyl, 2-chloro-2,2-difluoroethyl, 2,2-dichloro-2-fluoroethyl, 2,2,2-trichloroethyl, 2-bromo-2,2-difluoroethyl, 2-bromo-2-chloro-2-fluoroethyl, 2-bromo-2,2-dichloroethyl, 1,1,2,2-tetrafluoroethyl, pentafluoroethyl, 1-chloro-1,2,2,2-tetrafluoroethyl, 2-chloro-1,1,2,2-tetrafluoroethyl, 1,2-dichloro-1,2,2-trifluoroethyl, 2-bromo-1,1,2,2-tetrafluoroethyl, and the like.

[0349] The term "haloalkoxy" refers to an alkoxy group, as defined above, substituted with one or more halogen. Non-limiting examples include fluoromethoxy, chloromethoxy, bromomethoxy, iodomethoxy, difluoromethoxy, chlorofluoromethoxy, dichloromethoxy, bromofluoromethoxy, trifluoromethoxy, chlorodifluoromethoxy, dichlorofluoromethoxy, trichloromethoxy, bromodifluoromethoxy, bromochlorofluoromethoxy, dibromofluoromethoxy, and the like; preferably fluoromethoxy, difluoromethoxy, trifluoromethoxy, 2-fluoroethoxy, 2-chloroethoxy, 2-bromoethoxy, 2,2-difluoroethoxy, 2-chloro-2-fluoroethoxy, 2,2-dichloroethoxy, 2-bromo-2-fluoroethoxy, 2,2,2-trifluoroethoxy, 2-chloro-2,2-difluoroethoxy, 2,2-dichloro-2-fluoroethoxy, 2,2,2-trichloroethoxy, 2-bromo-2,2-difluoroethoxy, 2-bromo-2-chloro-2-fluoroethoxy, 2-bromo-2,2-dichloroethoxy, 1,1,2,2-tetrafluoroethoxy, pentafluoroethoxy, 1-chloro-1,2,2,2-tetrafluoroethoxy, 2-chloro-1,1,2,2-tetrafluoroethoxy, 1,2-dichloro-1,2,2-trifluoroethoxy, 2-bromo-1,1,2,2-tetrafluoroethoxy, preferably fluoromethoxy, difluoromethoxy, trifluoromethoxy, 2-fluoroethoxy, 2-chloroethoxy, 2-bromoethoxy, 2,2-difluoroethoxy.

[0350] The term "alkylidene" refers to a divalent, free radical alkyl structure formed by the loss of two hydrogen atoms, wherein alkyl is as defined above. Non-limiting examples include methylene ethylidene 1-methylethylidene

[0351] The term "haloalkyl" means an alkyl group substituted with one or more halogens, wherein alkyl is as defined above. Non-limiting examples include: fluoromethyl difluoromethyl

[0352] The term "cycloalkylalkyl" means an alkyl group substituted with one or more cycloalkyl groups, wherein alkyl is as defined above. Non-limiting examples include: cyclopropylmethyl cyclopentylmethyl cyclohexylmethyl

[0353] The term "heterocycloalkylalkyl" means an alkyl group substituted with one or more heterocycloalkyl groups, wherein alkyl is as defined above. Non-limiting examples include: oxetanyl azolidinyl

[0354] The term "cycloalkylalkyl" means an alkyl group substituted with one or more cycloalkyl groups, wherein alkyl is as defined above. Non-limiting examples include: cyclopropylmethyl cyclopentylmethyl

[0355] The term "(maleimide-N-yl)-" means The term "- (succinimid-3-yl-N)-" means

[0356] The term "mercapto" means -SH.

[0357] The term "hydroxy" means -OH.

[0358] The term "nitro" means -NO2.

[0359] The term "amino" means -NH2.

[0360] The term "cyano" means -CN.

[0361] The term "carboxy" means -C(O)OH.

[0362] The term "aldehyde" means -CHO.

[0363] The term "oxo" or "oxo group" means =O.

[0364] The term "carbonyl" means C=O.

[0365] The term "amido" means -C(O)NH2.

[0366] The term "sulfonyl" means -S(O)2.

[0367] The term "deuteroalkyl" means an alkyl group substituted with one or more deuterium, wherein alkyl is as defined above.

[0368] The term "deuteroalkoxy" means an alkoxy group substituted with one or more deuterium, wherein alkoxy is as defined above.

[0369] The term "haloalkoxy" means an alkoxy group substituted with one or more halogen, wherein alkoxy is as defined above.

[0370] The term "hydroxyalkyl" means an alkyl group substituted with one or more hydroxy, wherein alkyl is as defined above.

[0371] The term "alkylamino" means alkyl-NH-, wherein alkyl is as defined above.

[0372] The term "alkenylene" means a divalent straight and branched chain alkenyl group.

[0373] The term "alkynylene" means a divalent straight and branched chain alkynyl group.

[0374] The terms "comprise", "comprising", "have", "having", "include", "including", "contain", "containing", or "involve" and any variations thereof, are open-ended, and do not exclude additional, unrecited elements or method steps. It will be understood by those within the art that, in some instances, the terms "include", "comprise", and "have" can be used interchangeably.

[0375] The terms "one or more" or similar expressions "at least one" can mean, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more.

[0376] When a range of values is disclosed, any value, and any included range within the range, is specifically disclosed. In particular, each value within a range of values, and each range within a range of values, is specifically disclosed.

[0377] In the present text, "Z" and "-Z-" are both meant to represent the same specific group, which can be used interchangeably.

[0378] The expression m-n as used herein means the range from m to n and subranges and individual points within the range. For example, the expression "C2-C8" or "C2-C8" encompasses the range of 2 to 8 carbon atoms and is to be understood as also encompassing any sub-range within the range, as well as each individual point within the range, e.g., C2-C5, C3-C4, C2-C6, C3-C6, C4-C6, C4-C7, C4-C8, etc., as well as C2, C3, C4, C5, C6, C7, C8, etc. For example, the expression "C3-C6" encompasses the range of 3 to 6 carbon atoms and is to be understood as also encompassing any sub-range within the range, as well as each individual point within the range, e.g., C3-C4, C4-C5, C5-C6, C3-C5, C4-C6, C3-C6, C4-C6, C5-C6, etc., as well as C3, C4, C5, C6, etc. 2-8 The expression "C1-Cn" or "C1-Cn" as used herein means the range from 1 to n carbon atoms and is to be understood as also encompassing any sub-range within the range, as well as each individual point within the range, e.g., C1-C5, C2-C4, C3-C6, C4-C6, C4-C7, C4-C8, etc., as well as C1, C2, C3, C4, C5, C6, C7, C8, etc. For example, the expression "C1-C6" encompasses the range of 1 to 6 carbon atoms and is to be understood as also encompassing any sub-range within the range, as well as each individual point within the range, e.g., C1-C4, C1-C5, C1-C6, C2-C4, C2-C5, C2-C6, C3-C4, C3-C5, C3-C6, C4-C5, C4-C6, C5-C6, etc., as well as C1, C2, C3, C4, C5, C6, etc.10 "or "C 3-10 " should also be understood in like fashion, e.g., can encompass any sub-range and point value included therein, such as C3-C9, C6-C9, C6-C8, C6-C7, C7-C 10 , C7-C9, C7-C8, C8-C9, etc., and C3, C4, C5, C6, C7, C8, C9, C 10 , etc. For another example, the expression "C1-C6" or "C 1-6 " encompasses a range of 1 to 6 carbon atoms and should be interpreted to also cover any sub-range and individual integer point value therein, e.g., C2-C5, C3-C4, C1-C2, C1-C3, C1-C4, C1-C5, C1-C6, etc., as well as C1, C2, C3, C4, C5, C6, etc. For yet another example, the expression "from three to ten" should be interpreted to encompass any sub-range and individual integer point value therein, e.g., from three to five, from three to six, from three to seven, from three to eight, from four to five, from four to six, from four to seven, from four to eight, from five to seven, from five to eight, from six to seven, from six to eight, from nine to ten, etc., as well as three, four, five, six, seven, eight, nine, ten, etc. Other similar expressions are to be understood in like fashion herein.

[0379] The expressions "X is selected from A, B or C," "X is selected from A, B and C," "X is A, B or C," "X is A, B and C," and the like, are used interchangeably herein and express the same meaning, i.e., that X can be any one of or any combination of A, B, and C.

[0380] The term "optionally" or "optional" means that the subsequently described event or circumstance can or can not occur, and this description includes instances where the event or circumstance occurs and instances where it does not. For example, "cycloalkyl optionally substituted with alkyl" means that alkyl can or can not be present, and this description includes instances where the cycloalkyl is substituted with alkyl and instances where the cycloalkyl is not substituted with alkyl.

[0381] The terms "substituted" and "substitution" mean that one or more (e.g., one, two, three or four) hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution results in a stable compound. In addition, when a structural unit is substituted, even if the structural unit is noted to have a hydrogen atom, it does not mean that the hydrogen atom cannot be substituted, but rather that any position in the structural unit, including the hydrogen atom, can be substituted, e.g., a structural unit substituted means that any of the hydrogens on any of the positions, including the nitrogen atom, can be replaced by a substituent. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. By "stable compound" is meant a compound that possesses sufficient structural integrity for the compound to be synthesized and isolated. By "optionally substituted" is meant that the named atom is either unsubstituted or substituted with the named group(s). Exemplary substituents include, but are not limited to: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, C 5-12 aryl, 5-12 membered heteroaryl, -CO-(C 3-8 cycloalkyl), -CO-(3-8 membered heterocycloalkyl), -CO-(C 5-12 aryl), -CO-(5-12 membered heteroaryl), hydroxy, C 1-6 alkoxy, C 5-12 aryloxy, thiol, C 1-6 alkylthiol, cyano, halogen, oxo, aldehyde, SF5, SCF3, -N3, C 1-6 alkylthiocarbonyl, C 1-6 alkylcarbamoyl, N-carbamoyl, nitro, silyl, sulfinyl, sulfonyl, sulfoxide, carboxyl, C 1-6 haloalkyl, C 1-6 haloalkoxy, amino, phosphonic acid, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2, -OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 alkyl), -NHCO2(C 1-6 alkyl), -NHC(=O)N(C 1-6 alkyl)2, -HC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -CH=N(C 1-6 alkyl), -CH=N-O(C 1-6 alkyl), -NHSO2(C 1-6 alkyl), -SO2N(C 1-6 alkyl)2, -SO2NH(C 1-6alkyl), -SO2NH2, -SO2C 1-6 alkyl, etc.

[0382] If a substituent group is described as "optionally substituted" with a list of substituents, the substituent group can be unsubstituted or can be substituted with one or more of the optional substituents. If an atom or group is described as being optionally substituted with one or more of a list of substituents, then one or more hydrogens on the atom or group are each independently replaced with an optionally substituent. When the substituent is oxo (i.e., =0), it means that two hydrogen atoms are replaced. When the substituent is a ketal (e.g., ), it means that two hydrogen atoms are replaced. When the substituent is hydrogen, this can also mean that the corresponding group is "non-substituted" or "unsubstituted". Unless indicated, the point of attachment of a substituent group can be from any suitable position on the substituent group, as used herein.

[0383] When the bond to a substituent group is shown to be through a bond to a ring atom that connects two atoms in a ring, then such substituent group can be bonded to either ring-forming atom in the substitutable ring.

[0384] When any variable (e.g., R) occurs more than one time in a compound or substituent, its definition in each occurrence is independent of its definition at every other occurrence. For example, if a group is substituted with 0, 1, 2, 3, or 4 R substituents, then the group can optionally be substituted with up to four R substituents, and the selection of each R substituent is independent of the selection of each other R substituent.

[0385] When a linking group is recited without specifying the direction of attachment, then the direction of attachment includes both left-to-right and right-to-left reading orders, e.g., A-L-B, L selected from -M-W-, includes A-M-W-B and A-W-M-B, with A-M-W-B being preferred.

[0386] The compounds of the present application can exist in particular geometric or stereoisomeric forms. Stereoisomers are molecules that have the same atomic order but differ in the spatial arrangement of their atoms. All such compounds of the present application, including cis and trans isomers, optical isomers, and racemic mixtures and other mixtures thereof, are intended to be within the scope of the present application. The substituents of the compounds of the present application can have additional asymmetric carbon atoms. All such isomers, as well as mixtures thereof, are intended to be within the scope of the present application. In certain embodiments, preferred compounds are those isomers that exhibit greater biological activity. Purified or partially purified isomers and stereoisomers, or racemic mixtures or diastereomeric mixtures of the compounds of the present application are also within the scope of the present application. Purification and separation of such materials can be accomplished by standard techniques known in the art.

[0387] The compounds of the present application also include tautomeric forms thereof. Tautomers are functional groups that can be interconverted by a reversible chemical reaction, often involving the migration of a hydrogen atom and a pi bond (double or triple bond) and the conversion of one functional group to another. Examples of such pairs are aldehyde / keto-enol, imine-enamine.

[0388] Any hydrogen atom in the compounds of the present application can be replaced by its isotope deuterium.

[0389] The compounds of the present application include all suitable isotopic variations of the compounds. The term "isotopic variations" means the replacement of at least one atom with an atom having the same atomic number but an atomic mass different from the atomic mass of the atom that it replaces. Examples of isotopes that can be present in the compounds of the present disclosure include stable and radioactive isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulphur, fluorine, chlorine, bromine and iodine, such as 2 H (deuterium, D), 3 H (tritium, T), 11 C, 13 C, 14 C, 15 N, 17 O, 18 O, 32 P, 33 P, 33 S, 34 S, 35 S, 36 S, 18 F,36 Cl, 82 Br, 123 I, 124 I, 125 I, 129 I and 131 I, preferably deuterium.

[0390] Deuterated drugs have advantages of reducing side effects, increasing drug stability, enhancing efficacy, prolonging drug biological half-life, etc. compared with non-deuterated drugs. All isotopic composition changes of the compounds of the present disclosure, whether radioactive or not, are included within the scope of the present disclosure. Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom, wherein the replacement of deuterium can be partial or complete, and the partial replacement of deuterium refers to at least one hydrogen being replaced by at least one deuterium.

[0391] In the compounds of the present application, when a position is specifically designated as deuterium, D, the position is understood to have an abundance of deuterium that is at least 1000 times greater than the natural abundance of deuterium, which is 0.015% (i.e., at least 15% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 1000 times greater than the natural abundance of deuterium (i.e., at least 15% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 2000 times greater than the natural abundance of deuterium (i.e., at least 30% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 3000 times greater than the natural abundance of deuterium (i.e., at least 45% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 3340 times greater than the natural abundance of deuterium (i.e., at least 50.1% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 3500 times greater than the natural abundance of deuterium (i.e., at least 52.5% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 4000 times greater than the natural abundance of deuterium (i.e., at least 60% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 4500 times greater than the natural abundance of deuterium (i.e., at least 67.5% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 5000 times greater than the natural abundance of deuterium (i.e., at least 75% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 5500 times greater than the natural abundance of deuterium (i.e., at least 82.5% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 6000 times greater than the natural abundance of deuterium (i.e., at least 90% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 6333.3 times greater than the natural abundance of deuterium (i.e., at least 95% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 6466.7 times greater than the natural abundance of deuterium (i.e., at least 97% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 6600 times greater than the natural abundance of deuterium (i.e., at least 99% incorporation of deuterium). In some embodiments, the abundance of deuterium of each designated deuterium atom is at least 6633.3 times greater than the natural abundance of deuterium (i.e., at least 99.5% incorporation of deuterium).

[0392] The term "pharmaceutically acceptable" refers to a substance that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of a patient without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for its intended use.

[0393] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present application which is safe and effective for use in mammals, and possesses the desirable biological activity.

[0394] The term "pharmaceutical composition" refers to a composition comprising one or more compounds of the present application or physiologically / pharmaceutically acceptable salts or prodrugs thereof, and other components such as physiologically / pharmaceutically acceptable carriers or excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to a subject, and to facilitate absorption of the active ingredient to exert a biological activity.

[0395] The term "pharmaceutically acceptable carrier" refers to those agents that do not cause significant irritation to an organism and do not abrogate the biological activity and properties of the active compound. "Pharmaceutically acceptable carriers" include, but are not limited to, glidants, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersing agents, effervescent agents, stabilizers, solvents, or emulsors.

[0396] The terms "administration" or "administering" or the like refer to methods allowing the delivery of a compound or composition to the desired site of biological action. These methods include, but are not limited to, oral or parenteral (including intracerebroventricular, intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular injection or infusion), topical, rectal administration, and the like. In particular, injection or oral administration.

[0397] As used herein, the term "treatment" includes alleviating, abating or ameliorating a disease or condition, preventing the onset of other symptoms, improving or preventing the underlying metabolic factors causing symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, and extending to prophylaxis. "Treatment" also includes achieving a therapeutic benefit and / or a prophylactic benefit. By therapeutic benefit is meant eradication or amelioration of symptoms, and / or underlying metabolic causes of symptoms, although amelioration can not be complete. Also, a therapeutic benefit is achieved with the eradication or substantial diminution of any physiological parameter associated with a disease or condition, even though the patient can still be affected by the disease or condition. By prophylactic benefit is meant prevention of disease or condition, or prevention of a decrease in quality of life associated with a disease or condition.

[0398] The term "active ingredient," "therapeutic agent," "active substance," or "active agent" refers to a chemical entity that is effective in treating or preventing a disorder, disease, or condition of interest. The term "neuropsychiatric disorder" refers to the general class of neurological and psychiatric disorders, including neurological and / or psychiatric disorders.

[0399] The term "effective amount," "therapeutically effective amount," or "prophylactically effective amount" in reference to a drug, drug unit, or active ingredient means an amount of the drug or agent that is acceptable in terms of side effects, but that is sufficient to achieve the intended result. The determination of an effective amount is dependent on the age and general condition of the individual, as well as the particular active substance, and an appropriate effective amount in a given case can be determined by a person skilled in the art according to routine experiments.

[0400] As used herein, "individual" includes a human or non-human animal. Exemplary human individuals include a human individual (referred to as a patient) who has a disease (e.g., a disease described herein) or a normal individual. "Non-human animals" in the present application include all vertebrates, e.g., non-mammals (e.g., birds, amphibians, reptiles), and mammals, e.g., non-human primates, domestic animals, and / or laboratory models, (e.g., sheep, dog, cat, cow, pig, etc.).

[0401] The term "antibody drug conjugate" (ADC) refers to a monoclonal antibody or antibody fragment linked to a toxic drug with biological activity through a stable linker unit.

[0402] The term "antibody" refers to an immunoglobulin, which is a four polypeptide chain structure connected by interchain disulfide bonds, consisting of two identical heavy chains and two identical light chains. The antigenicity of immunoglobulin heavy chain constant region is different due to different amino acid composition and arrangement order. Accordingly, immunoglobulin can be divided into five types, or called isotypes of immunoglobulin, namely IgM, IgD, IgG, IgA and IgE, and the corresponding heavy chains are μ chain, δ chain, γ chain, α chain and ε chain, respectively. The same type of Ig can be divided into different subtypes according to the difference of amino acid composition of hinge region and the number and position of heavy chain disulfide bonds, such as IgG can be divided into IgG1, IgG2, IgG3 and IgG4. The light chain is divided into κ chain or λ chain through the constant region. Each of the five types of Ig can have κ chain or λ chain. The antibody described in the present application is preferably a specific antibody against a cell surface antigen on a target cell.

[0403] The term "room temperature" refers to a temperature from 10 °C to 40 °C. In some embodiments, "room temperature" refers to a temperature from 15 °C to 30 °C; in other embodiments, "room temperature" refers to a temperature from 18 °C to 25 °C.

[0404] "Equivalent" or its abbreviation "eq" is based on the equivalent relationship of chemical reactions, with the basic raw material used in each step as the basis (1 equivalent), and the equivalent amount of other raw materials required.

[0405] The following detailed description of the application is intended to illustrate, but not limit, the embodiments of the application, to enable a person skilled in the art to better understand the technical solutions of the application, its principles and its practical applications, so that the application can be modified and implemented in many forms by those skilled in the art to best meet the requirements of specific uses. BRIEF DESCRIPTION OF DRAWINGS

[0406] Figure 1: 293T+293T-CDH6 transport well plate co-incubation test results.

[0407] Figure 2: OVCAR3 tumor growth curve. DETAILED DESCRIPTION

[0408] The content of the application will be described in detail below through examples. If the specific conditions are not specified in the examples, the experimental methods are carried out according to the conventional conditions. The examples are given to better illustrate the content of the application, but it should not be understood as limiting the content of the application to the examples. Those skilled in the art can make non-essential improvements and adjustments to the embodiments according to the above content of the application, which still belong to the protection scope of the application.

[0409] The structure of the compound is determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS). The NMR shift (δ) is given in units of 10 -6 (ppm). The NMR is measured by a nuclear magnetic instrument (Bruker Avance III 400 and Bruker Avance 300), the measuring solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS);

[0410] The MS is measured by (Agilent 6120B (ESI) and Agilent 6120B (APCI));

[0411] The HPLC is measured by using an Agilent 1260DAD high-pressure liquid chromatograph (Zorbax SB-C 18 100×4.6mm, 3.5μM);

[0412] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography is 0.15mm-0.20mm, and the specification of the product used in thin layer chromatography separation and purification is 0.4mm-0.5mm;

[0413] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier;

[0414] DCM: dichloromethane;

[0415] DMF: N,N-dimethylformamide;

[0416] DIEA: N,N-diisopropylethylamine;

[0417] MeOH: methanol;

[0418] TFA: trifluoroacetic acid;

[0419] DMSO: dimethyl sulfoxide;

[0420] DIC: N,N'-diisopropylcarbodiimide;

[0421] HOBT: 1-hydroxybenzotriazole;

[0422] HOAT: N-hydroxy-7-azabenzotriazole.

[0423] Examples

[0424] The embodiments of the present application will be described in detail below with examples, but those skilled in the art will understand that the following examples are only for illustration of the present application and should not be regarded as limiting the scope of the present application. The specific conditions not noted in the examples are carried out according to the conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used are not noted the manufacturer, which are all the conventional products that can be obtained by market purchase. The proportions or percentages used in this text are by weight, unless otherwise specified.

[0425] 1. Synthesis of toxin

[0426] Example 1:

[0427] First step: 1A (5.00 g, 39.0 mmol) was dissolved in a mixed solvent of dichloromethane (180 mL) and methanol (20 mL), and trimethylsilyldiazomethane (29.3 mL, 58.5 mmol, 2N in Hexane) was added dropwise at 0°C. After being stirred at room temperature, the reaction was monitored by TLC and was directly dried. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 80:20) to obtain compound 1B (4.90 g, 88.3%).

[0428] LC-MS (ESI): m / z = 143.1 [M+H] + .

[0429] Second step: Methyl triphenylphosphonium bromide (18.8 g, 52.8 mmol) was dissolved in dry THF (200 mL), and n-butyllithium solution (19.7 mL, 49.2 mmol, 2.5 N in THF) was added dropwise at 0 °C. After the addition was completed, the reaction was continued for 1 h. Compound 1B (5.00 g, 35.2 mmol) was dissolved in dry THF (20 mL), and the mixture was gradually warmed to room temperature and reacted overnight. After the completion of the reaction was monitored by TLC, saturated aqueous ammonium chloride solution (200 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 5). The combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure at low temperature. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 90:10) to give compound 1C (3.02 g, 61.2%).

[0430] LC-MS (ESI): m / z = 141.1 [M+H] + .

[0431] Third step: Compound 1C (1.00 g, 7.13 mmol) was dissolved in dry THF (50 mL), and KHMDS solution (10.7 mL, 10.7 mmol, 1.0 N in THF) was added dropwise at -78 °C. After the addition was completed, the reaction was continued for 1 h. Davis Oxaziridine (2.24 g, 8.56 mmol) was dissolved in dry THF (10 mL), and the mixture was reacted while maintaining the low temperature. After the completion of the reaction was monitored by TLC, saturated aqueous ammonium chloride solution (200 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 5). 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 (v:v) = 100:0 ~ 85:15) to give compound 1D (500 mg, 44.8%).

[0432] LC-MS (ESI): m / z = 157.1 [M+H] + .

[0433] Fourth step: Compound 1D (100 mg, 0.64 mmol) was dissolved in a mixed solvent of tetrahydrofuran (2 mL), methanol (2 mL), and water (1 mL), and lithium hydroxide monohydrate (107 mg, 2.56 mmol) was added at room temperature. After the addition was completed, the reaction was continued. After the completion of the reaction was monitored by TLC, the mixture was directly concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol (v:v) = 100:0 ~ 66:34) to give compound 1E (56 mg, 61.5%).

[0434] LC-MS (ESI): m / z = 141.1 [M-H] - .

[0435] Step 5: Compound 1E (56 mg, 0.394 mmol), mesylate of compound 1F (209 mg, 0.394 mmol), HATU (300 mg, 0.788 mmol), HOBT (106 mg, 0.788 mmol) and DIPEA (254 mg, 1.97 mmol) were dissolved in dry N,N-dimethylformamide (20 mL) and reacted at room temperature overnight. After monitoring the completion of the reaction by TLC, saturated aqueous ammonium chloride solution (50 mL) was added, extracted with ethyl acetate (20 mL x 5), the organic phase was combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 20:80) to obtain a mixture of compound 1-1 and 1-2 (73 mg, 33.1%), which was separated by SFC to obtain two isomers, compound 1-1 (10 mg, retention time 1.505 min, 4.54%) and compound 1-2 (4 mg, retention time 1.073 min, 1.81%).

[0436] Preparative chromatography separation conditions: 1. Instrument: Waters 150Prep-SFC F; 2. Column: Chiral IC column; 3. Mobile phase system: A carbon dioxide; B ethanol and acetonitrile; 4. Gradient: B 45%; 5. Flow rate: 100 mL / min.

[0437] Compound 1-1:

[0438] 1 H NMR (400 MHz, Chloroform-d) d 7.66 (d, 1H), 7.57 (s, 1H), 7.23 (s, 1H), 5.62 (d, 1H), 5.58-5.49 (m, 1H), 5.26-5.07 (m, 3H), 4.78 (d, 2H), 4.24 (d, 1H), 3.16 (t, 2H), 2.95-2.72 (m, 4H), 2.67 (d, 1H), 2.43 (s, 3H), 2.39-2.19 (m, 2H), 1.95-1.81 (m, 2H), 1.26 (s, 3H).

[0439] LC-MS (ESI): m / z = 560.3 [M+H] + .

[0440] Compound 1-2:

[0441] 1H NMR (400 MHz, Chloroform-d) δ 7.52 - 7.39 (m, 3H), 5.66 (d, 1H), 5.39 - 5.29 (m, 1H), 5.23 (d, 1H), 4.81 (d, 2H), 4.76 - 4.62 (m, 2H), 4.42 (d, 1H), 3.21 (d, 1H), 3.14 - 3.03 (m, 1H), 2.97 - 2.66 (m, 5H), 2.47 - 2.28 (m, 4H), 2.10 - 2.02 (m, 1H), 1.84 - 1.75 (m, 2H), 1.26 (s, 3H).

[0442] LC-MS (ESI): m / z = 560.3 [M+H] + .

[0443] Example 2

[0444] First Step: Take 2A (10.0 g, 54.35 mmol) as raw material, refer to the synthetic method of example 1 second step to obtain compound 2B (7.0 g, 70%).

[0445] 1 H NMR (400 MHz, CDC13) δ 4.66 - 4.49 (m, 2H), 4.19 - 4.07 (m, 2H), 2.35 - 2.25 (m, 2H), 2.23 - 2.17 (m, 2H), 2.10 - 1.98 (m, 2H), 1.99 - 1.89 (m, 1H), 1.90 - 1.77 (m, 2H), 1.31 - 1.20 (m, 3H), 1.15 - 1.02 (m, 2H).

[0446] Second Step: Take 2B (5.0 g, 27.47 mmol) as raw material, refer to the synthetic method of example 1 third step to obtain compound 2C (0.5 g, 9%).

[0447] LC-MS (ESI): m / z = 199.1 [M+H] + .

[0448] Third Step: Take 2C (0.3 g, 1.52 mmol) as raw material, refer to the synthetic method of example 1 fourth step to obtain compound 2D (0.2 g, 77%).

[0449] LC-MS (ESI): m / z = 169.1 [M-H] - .

[0450] Fourth step: 2D (0.1 g, 0.59 mmol) was dissolved in N,N-dimethylformamide (3 mL), PyBOP (307 mg, 0.59 mmol), compound 1F (314 mg, 0.59 mmol) and DIPEA (380 mg, 2.95 mmol) were added and reacted for 16 hours. After the reaction was completed, water (30 mL) was added to the reaction solution, extracted with ethyl acetate (15 mL x 3), the organic layers were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was purified by high performance liquid chromatography to obtain compound 2-1 (45 mg, retention time 4.609 min, yield 13%) and compound 2-2 (48 mg, retention time 4.845 min, yield 14%).

[0451] Analytical HPLC method: 1. Instrument: Shimadzu LC-20AT; 2. Column: Xtimate C18 4.6*50mm, 3um; 3. Mobile phase A: 0.05% TFA solution, mobile phase B: acetonitrile; 4. Gradient: A 95-5% B 5-95%; 5. Flow rate: 1 mL / min, column temperature: 35°C, wavelength: 210nm / 254nm, collection time: 10min.

[0452] Preparation method: Instrument: waters 2767 preparative liquid phase; column: SunFire@Prep C18(19mm*250mm). The sample was dissolved in methanol and filtered with a 0.22um filter to prepare a sample solution. The preparation chromatographic conditions: mobile phase A, B composition: mobile phase A: acetonitrile, mobile phase B: water (containing 1% ammonium acetate), gradient elution, mobile phase A content from 10%-40%, flow rate 12mL / min. Elution time 25min.

[0453] Compound 2-1: LC-MS (ESI): m / z = 588.2 [M+H] + .

[0454] 1 H NMR (400 MHz, DMSO-d6) δ 8.39-8.25 (m, 1H), 7.85-7.69 (m, 1H), 7.30 (s, 1H), 6.48 (s, 1H), 5.59-5.46 (m, 1H), 5.42 (s, 2H), 5.39-5.28 (m, 1H), 5.24-5.12 (m, 2H), 4.58 (s, 2H), 3.84-3.68 (m, 1H), 3.22-3.07 (m, 2H), 2.39 (s, 3H), 2.31-2.23 (m, 2H), 2.19-1.68 (m, 9H), 1.30-1.17 (m, 2H), 0.92-0.79 (m, 3H).

[0455] Compound 2-2: LC-MS (ESI): m / z = 588.2 [M+H] + .

[0456] 1 H NMR (400 MHz, DMSO-d6) δ 8.37 - 8.24 (m, 1H), 7.79 - 7.64 (m, 1H), 7.29 (s, 1H), 6.49 (s, 1H), 5.55 - 5.48 (m, 1H), 5.46 - 5.39 (m, 3H), 5.23 - 5.01 (m, 2H), 4.58 (s, 2H), 3.93 - 3.85 (m, 1H), 3.21 - 3.04 (m, 2H), 2.36 (s, 3H), 2.31 - 1.83 (m, 9H), 1.77 - 1.61 (m, 2H), 1.33 - 1.18 (m, 2H), 0.96 - 0.76 (m, 3H).

[0457] Example 3

[0458] First step: potassium tert-butoxide (10.9 g, 97.83 mmol) was dissolved in tetrahydrofuran (100 mL), and a mixture of 3A (10.0 g, 54.35 mmol) and 2-(difluoromethyl)sulfonyl)pyridine (11.5 g, 59.8 mmol) in tetrahydrofuran (50 mL) was added dropwise at -40 °C, and the reaction was allowed to warm to room temperature overnight. Saturated ammonium chloride (65 mL) and hydrochloric acid (3N, 65 mL) were added, and stirring was continued at room temperature for 3 h. Ethyl acetate (300 mL x 1) was added, and the combined organic phases were washed with saturated brine (300 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated. Silica gel column chromatography of the residue gave compound 3B (7.5 g, 63%).

[0459] 1 H NMR (400 MHz, CDCl3) δ 4.20 - 4.03 (m, 2H), 2.49 - 2.39 (m, 2H), 2.25 - 2.17 (m, 2H), 1.94 - 1.74 (m, 5H), 1.29 - 1.21 (m, 3H), 1.12 - 0.98 (m, 2H).

[0460] Second step: compound 3C (0.25 g, 4%) was obtained by using 3B (5.0 g, 22.9 mmol) as the raw material, according to the synthetic method of Example 1, third step.

[0461] LC-MS (ESI): m / z = 235.1 [M+H] + .

[0462] Third Step: Take 3C (0.25 g, 1.06 mmol) as raw material, and refer to the synthetic method of Example 1 fourth step to obtain compound 3D (0.2 g, 91%).

[0463] LC-MS (ESI): m / z = 205.1 [M-H] - .

[0464] Fourth Step: Take 3D (0.1 g, 0.49 mmol) as raw material, and refer to the synthetic method of Example 2 fourth step to obtain compound 3-1 (50 mg, retention time 4.744 min, yield 16%) and compound 3-2 (55 mg, retention time 4.979 min, yield 18%).

[0465] Analytical HPLC method: 1. Instrument: Shimadzu LC-20AT; 2. Column: Xtimate C18 4.6*50mm, 3μm; 3. Mobile phase A: 0.05% TFA solution, mobile phase B: acetonitrile; 4. Gradient: A 95-5% B 5-95%; 5. Flow rate: 1 mL / min, column temperature: 35℃, wavelength: 210nm / 254nm, collection time: 10min.

[0466] Preparation method: Instrument: waters 2767 preparative liquid phase; column: SunFire@Prep C18(19mm*250mm). The sample was dissolved with methanol and filtered with a 0.22μm filter to prepare a sample solution. The preparation chromatographic conditions: mobile phase A, B composition: mobile phase A: acetonitrile, mobile phase B: water (containing 1% ammonium acetate), gradient elution, mobile phase A content from 10%-40%, flow rate 12mL / min. Elution time 25min.

[0467] Compound 3-1: LC-MS (ESI): m / z = 624.2 [M+H] + .

[0468] 1 H NMR (400 MHz, DMSO-d6) δ 8.45-8.32 (m, 1H), 7.82-7.70 (m, 1H), 7.30 (s, 1H), 6.49 (s, 1H), 5.58-5.49 (m, 1H), 5.48-5.30 (m, 3H), 5.22-5.13 (m, 2H), 3.82-3.76 (m, 1H), 3.19-3.10 (m, 2H), 2.45-2.36 (m, 5H), 2.19-2.10 (m, 2H), 1.92-1.72 (m, 7H), 1.27-1.19 (m, 2H), 0.93-0.83 (m, 3H).

[0469] Compound 3-2: LC-MS (ESI): m / z = 624.2 [M+H] + .

[0470] 1 H NMR (400 MHz, DMSO-d6) δ 8.38 - 8.29 (m, 1H), 7.78 - 7.68 (m, 1H), 7.30 (s, 1H), 6.50 (s, 1H), 5.58 - 5.51 (m, 1H), 5.51 - 5.47 (m, 1H), 5.42 (s, 2H), 5.29 - 5.06 (m, 2H), 3.96 - 3.85 (m, 1H), 3.20 - 3.09 (m, 2H), 2.45 - 2.35 (m, 5H), 2.25 - 2.08 (m, 2H), 1.92 - 1.67 (m, 7H), 1.31 - 1.19 (m, 2H), 0.91 - 0.82 (m, 3H).

[0471] Example 4

[0472] First step: Compound 4A (11 g, 85.87 mmol), diiodomethane (34.5 g, 128.81 mmol) and potassium carbonate (17.8 g, 128.81 mmol) were dissolved in N,N-dimethylformamide (100 mL), after the addition was completed, the reaction was stirred at 100 °C for 3 hours. Concentrated under reduced pressure, diluted with water (200 mL), extracted with ethyl acetate (100 mL) twice, the combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, the obtained residue was purified by column chromatography (petroleum ether) to obtain compound 4B (8.7 g, yield: 72.3%).

[0473] 1 H NMR (400 MHz, CDCl3) δ 6.78 - 6.73 (m, 1H), 6.68 - 6.63 (m, 2H), 5.99 (s, 2H).

[0474] Second step: Compound 4B (8.7 g, 62.09 mmol) was dissolved in anhydrous methanol (150 mL), NBS (8.84 g, 49.67 mmol) was added in batches, after the addition was completed, the reaction was stirred at 70 °C for 1 hour. The reaction solution was concentrated under reduced pressure, diluted with water (150 mL), extracted with ethyl acetate (80 mL) twice, the combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, the obtained residue was purified by column chromatography (petroleum ether: ethyl acetate (v:v) = 80:1) to obtain compound 4C (8.4 g, yield: 61.7%).

[0475] 1H NMR (400 MHz, DMSO-d6) δ 7.16-7.13 (m, 1H), 6.82-6.80 (m, 1H), 6.17 (s, 2H).

[0476] Third step: 4C (8.2 g, 37.44 mmol), N-BOC amine (13.16 g, 112.32 mmol), cesium carbonate (36.6 g, 112.32 mmol) and Xphos Pd G2 (2.95 g, 3.74 mmol) were added into the reaction flask in turn, 1,4-dioxane (160 mL), replaced with nitrogen for 3 times, stirred at 100 °C overnight. Filtration, the filtrate was concentrated under reduced pressure, the obtained residue was purified by column chromatography (petroleum ether: ethyl acetate (v:v) = 20:1) to obtain compound 4D (6.0 g, yield: 62.7%).

[0477] LC-MS (ESI): m / z = 200.0 [M-55] + .

[0478] Fourth step: Compound 4D (2.28 g, 8.93 mmol) was dissolved in dichloromethane (30 mL), trifluoroacetic acid (6 mL) was added, and the reaction was stirred at room temperature for 2 hours. Concentrated under reduced pressure to obtain the crude product, diluted with water (50 mL), the aqueous phase was extracted with petroleum ether twice to remove impurities, then adjusted to basic with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate (30 mL) twice, combined the organic phase, dried with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 4E (1.1 g, yield: 79.3%).

[0479] LC-MS (ESI): m / z = 156.1 [M+1] + .

[0480] Fifth step: Compound 4E (1.2 g, 7.74 mmol) was dissolved in 1,2-dichloroethane (30 mL), replaced with nitrogen for 3 times, cooled to 0 °C, BCl3 (1 M, 9.29 mL, 9.29 mmol) was added slowly dropwise, then 4-chlorobutyronitrile (0.96 g, 9.29 mmol) was added, after the addition was completed, the temperature was raised to 80 °C and the reaction was carried out for 2 hours. Cooled to 0 °C, 2M HCl (7.2 mL) was added slowly dropwise, after the addition was completed, the temperature was raised to 80 °C and the reaction was carried out for 0.5 hours. Diluted with water (50 mL), extracted with dichloromethane (50 mL) twice, combined the organic phase, dried with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the residue, which was purified by column chromatography (petroleum ether: ethyl acetate (v:v) = 20:1) to obtain compound 4F (0.38 g, yield: 18.92%).

[0481] LC-MS (ESI): m / z = 260.1 [M+1] + .

[0482] Sixth step: To the reaction flask was added 4F (0.38 g, 1.46 mmol), MeCN (10 mL), H20 (10 mL) and potassium carbonate (0.3 g, 2.19 mmol) successively, after the addition was completed, the reaction was stirred at 50 °C overnight. Concentrated under reduced pressure, diluted with water (30 mL), extracted with ethyl acetate (30 mL) twice, combined the organic phase, dried over anhydrous sodium sulfate and concentrated under reduced pressure, the obtained residue was purified by column chromatography (petroleum ether: ethyl acetate (v:v) = 1:1) to give compound 4G (0.3 g, yield: 84.9%).

[0483] LC-MS (ESI): m / z = 242.1 [M+1] + .

[0484] Seventh step: To the reaction flask was added 4G (73 mg, 0.30 mmol), 4H (200 mg, 0.76 mmol), p-toluenesulfonic acid (68 mg, 0.4 mmol) and N-methylpyrrolidone (8 mL) successively under nitrogen protection, after the addition was completed, the reaction was stirred at 110 °C for 30 minutes, then 4G (55 mg, 0.23 mmol) was added every 30 minutes for a total of 3 times. Diluted with water (30 mL), extracted with dichloromethane / methanol (10 / 1, 30 mL) three times, combined the organic phase, dried over anhydrous sodium sulfate and concentrated under reduced pressure, the obtained residue was purified by column chromatography (dichloromethane:methanol (v:v) = 10:1) and preparative-HPLC to give compound 4 (40 mg, yield: 8.58%).

[0485] 1 H NMR (400 MHz, DMSO-d6) δ 7.53 (s, 1H), 7.26 (s, 1H), 6.48 (s, 1H), 6.36 (s, 2H), 5.42 (s, 2H), 5.28 (s, 2H), 4.66-4.64 (m, 1H), 3.49-3.46 (m, 2H), 3.17-3.13 (m, 2H), 1.88-1.79 (m, 4H), 0.90-0.86 (m, 3H);

[0486] LC-MS (ESI): m / z = 469.1 [M+H] + .

[0487] Example 5

[0488] First Step: Compound 5A (2.0 g, 8.29 mmol) was taken in a reaction bottle, then hydrochloric acid 1,4-dioxane solution (10 ml) was added and allowed to react for 1 h at room temperature. After completion of the reaction as monitored by TLC, petroleum ether (50 ml) was added, stirred for 10 min, filtered, and the filter cake was collected and dried to get compound 5B (1.4 g, yield: 95.04%).

[0489] LC-MS (ESI): m / z = 142.1 [M+H] + .

[0490] Second Step: Compound 5B (600 mg, 3.38 mmol), glycolic acid (257 mg, 3.38 mmol) and DIPEA (2.94 ml, 16.9 mmol) were taken in a reaction bottle, dissolved in DMF (10 ml), then HATU (1.54 g, 4.05 mmol) was added and allowed to react for 16 h at room temperature. After completion of the reaction as monitored by TLC, water (100 ml) was added for dilution. Then it was extracted with ethyl acetate twice (100 ml x 2), the organic phase was combined and dried, concentrated under reduced pressure, and the residue obtained was purified by silica gel column chromatography to get compound 5C (330 mg, yield: 49.03%).

[0491] LC-MS (ESI): m / z = 200.2 [M+H] + .

[0492] Third Step: Compound 5C (300 mg, 1.50 mmol) was taken in a reaction bottle, dissolved in THF:MeOH:H2O = 3:1:1 (10 ml), then lithium hydroxide (144 mg, 6.00 mmol) was added and allowed to react for 4 h at room temperature. After completion of the reaction as monitored by TLC, pH was adjusted to 4-5, concentrated under reduced pressure, and the residue obtained was purified by silica gel column chromatography to get compound 5D (150 mg, yield: 54.01%).

[0493] LC-MS (ESI): m / z = 186.1 [M+H] + .

[0494] Fourth Step: Irinotecan mesylate (100 mg, 0.23 mmol), compound 5D (130 mg, 0.69 mmol), HOBT (31 mg, 0.23 mmol) and DIPEA (0.4 ml, 2.3 mmol) were taken in a reaction bottle, dissolved in DMF (10 ml), then EDCI (66 mg, 0.35 mmol) was added and allowed to react for 16 h at room temperature. After completion of the reaction as monitored by TLC, it was purified by reverse phase column to get compound 5 (30 mg, yield: 21.64%).

[0495] 1H NMR (400 MHz, DMSO-d6) δ 8.74-8.51 (m, 1H), 7.83-7.77 (m, 1H), 7.33-7.31 (m, 1H), 6.65-6.29 (m, 1H), 5.56-5.46 (m, 1H), 5.43 (s, 2H), 5.29-5.17 (m, 2H), 5.06-4.99 (m, 2H), 4.59-4.52 (m, 1H), 4.18-4.09 (m, 2H), 4.08-3.97 (m, 2H), 3.86-3.79 (m, 1H), 3.21-3.13 (m, 2H), 2.99-2.79 (m, 1H), 2.69-2.56 (m, 1H), 2.41 (s, 3H), 2.20-2.06 (m, 2H), 1.93-1.83 (m, 2H), 0.92-0.86 (m, 3H).

[0496] LC-MS (ESI): m / z = 603.2 [M+H] + .

[0497] Example 6:

[0498] First step: Compound 6A (5 g, 20.72 mmol) was dissolved in tetrahydrofuran (50 mL), diisobutylaluminum hydride (20.7 mL, 20.72 mmol, 1M) was added dropwise at -78°C, and the reaction was continued for 16 hours after the dropwise addition was completed. Methanol (5 ml) was added for quenching, water (100 ml) was added, ethyl acetate (40 mL x 3) was extracted, the organic phase was combined, dried with anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 1:0 ~ 1:1) to obtain compound 6B (3 g, yield: 72.66%).

[0499] LC-MS (ESI): m / z = 200.1 [M+H] + .

[0500] Second step: Compound 6B (350 mg, 1.76 mmol) was dissolved in dichloromethane (5 mL), followed by the addition of trifluoroacetic acid (2 ml), and the reaction was carried out at room temperature for 2 hours. Then the crude compound 6C (540 mg) was directly concentrated under reduced pressure.

[0501] LC-MS (ESI): m / z = 100.1 [M+H] + .

[0502] Third step: The mesylate salt of compound 1F (450 mg, 0.85 mmol), N,N- diisopropylethylamine (219 mg, 1.70 mmol) and N,N'-carbonyldiimidazole (138 mg, 0.85 mmol) were dissolved in dry N,N-dimethylformamide (20 mL) and reacted at room temperature for 3 hours. Then crude compound 6C (540 mg, 1.7 mmol) was added and the reaction was continued at room temperature for 16 hours. Water (50 ml) was added and the product was extracted with ethyl acetate (30 mL x 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the residue obtained was subjected to preparative HPLC to obtain compound 6 (68 mg, yield: 14.33%).

[0503] LC-MS (ESI): m / z = 561.3 [M+H] + .

[0504] 1 H NMR (400 MHz, DMSO-d6) δ 7.76 (d, 1H), 7.31 (s, 1H), 7.03 (d, 1H), 6.49 (s, 1H), 5.42 (s, 3H), 5.36 (q, 1H), 5.22 (s, 2H), 4.65 (s, 1H), 4.50 (s, 2H), 4.42 (s, 2H), 3.88 (s, 2H), 3.23-3.09 (m, 2H), 2.39 (s, 3H), 2.14 (q, 2H), 1.90-1.83 (m, 2H), 0.87 (t, 3H).

[0505] Example 7:

[0506] First step: 7A (1.50 g, 6.20 mmol) was dissolved in dry DMF (100 mL), and cesium carbonate powder (4.04 g, 12.40 mmol) was added at 0°C. Iodomethane (4.40 g, 31.0 mmol) was added dropwise with stirring, and the reaction was slowly warmed to room temperature. The reaction was monitored by TLC, and when the reaction was complete, the reaction solution was poured into 250 mL of ice water, stirred, and extracted with 200 mL of ethyl acetate. The organic phase was washed sequentially with water (200 mL x 2) and saturated sodium chloride (100 mL x 1), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 to 95:5) to obtain compound 7B (1.00 g, 63.0%).

[0507] LC-MS (ESI): m / z = 256.0 [M+H] + .

[0508] Second step: To a solution of 7B (0.50 g, 1.95 mmol) in anhydrous N-methyl pyrrolidine (5 mL), cuprous iodide (0.011 g, 0.058 mmol), dichlorobis(triphenylphosphine)palladium (0.041 g, 0.058 mmol), triethylamine (1.97 g, 19.5 mmol) were added successively, and the reaction system was purged with nitrogen for 3 times. 3-Butyn-1-ol (0.41 g, 5.85 mmol) was added dropwise rapidly at 70 °C. After the reaction was completed, the reaction solution was poured into 300 mL of an ammonia solution (5%) and stirred to precipitate a solid. The solid was filtered under suction, and the obtained solid was stirred in 200 mL of water again, filtered under suction, and dried under reduced pressure to obtain compound 7C (0.385 g, 80.4%).

[0509] LC-MS (ESI): m / z = 246.1 [M+H] + .

[0510] Third step: Compound 7C (0.40 g, 1.63 mmol) was dissolved in a mixture of ethanol (5 mL) and water (0.55 mL), and tin powder (0.39 g, 3.26 mmol), sodium sulfide nonahydrate (0.12 g, 0.49 mmol), concentrated hydrochloric acid (1.38 mL, 16.3 mmol) were added successively, and the reaction system was purged with nitrogen for 3 times. The reaction was stirred at 80 °C. After the reaction was completed, the reaction system was cooled to room temperature, and the reaction solution was diluted with ethyl acetate (100 mL), poured into 10% sodium bicarbonate solution (250 mL) and stirred for 10 min, and then allowed to stand to separate into two phases. The organic phase was washed successively with water (200 mL x 2), saturated sodium chloride (100 mL x 1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 to 30:70) to obtain compound 7D (0.22 g, 57.8%).

[0511] LC-MS (ESI): m / z = 234.1 [M+H] + .

[0512] Fourth step: Compound 7D (0.18 g, 0.77 mmol) and 4H (0.24 g, 0.92 mmol) were dissolved in dry toluene (5 mL), and 4-methylbenzenesulfonic acid pyridine (0.097 g, 0.39 mmol) was added at room temperature. After the addition was completed, the reaction system was purged with nitrogen and heated to 110 °C. After the reaction was completed, the reaction system was directly concentrated under reduced pressure, and the residue was purified by high performance liquid preparation to obtain compound 7 (2.45 mg, retention time 24.0 min, yield 0.69%).

[0513] Preparation method: instrument: waters 2767 preparative liquid phase; column: SunFire@Prep C18(19 mm x 250 mm). The sample was dissolved with DMF and filtered with a 0.45 μm filter to prepare a sample solution. The preparation chromatographic conditions: mobile phase A, B composition: mobile phase A: acetonitrile mobile phase B: water (5 mmol ammonium acetate), gradient elution, mobile phase B content from 5% to 40%, flow rate 15 mL / min. Elution time 25 min.

[0514] LC-MS (ESI): m / z = 461.1 [M+H] + .

[0515] 1 H NMR (400 MHz, DMSO-d6) δ 8.74 (s, 1H), 8.16 (d, 1H), 8.05 (d, 1H), 7.30 (s, 1H), 6.49 (s, 1H), 5.43 (s, 2H), 5.35 (s, 2H), 4.90 (s, 1H), 4.21 (s, 3H), 3.65-3.63 (m, 2H), 1.95-1.86 (m, 4H), 1.23 (s, 2H), 0.90 (t, 3H).

[0516] Example 8:

[0517] First step: potassium tert-butoxide (7.11 g, 63.4 mmol) was dissolved in dry THF (200 mL) and cooled to -40 °C, while dropping a solution of difluoromethyl(2-pyridyl)sulfone (7.48 g, 38.7 mmol) in THF (50 mL) and a solution of compound IB (5.00 g, 35.2 mmol) in THF (30 mL) were added. The temperature was allowed to rise to room temperature and the reaction was left overnight. After TLC monitoring of the disappearance of the starting material, saturated aqueous ammonium chloride (40 mL) and 3 N dilute hydrochloric acid (40 mL) were added and stirring was continued for about 4 hours. After adding a small amount of water, extraction was performed with ethyl acetate (200 mL x 2), the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure at low temperature and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 90:10) to obtain compound 8A (2.18 g, 35.2%).

[0518] LC-MS (ESI): m / z = 177.1 [M+H] + .

[0519] Second Step: Compound 8A (600 mg, 3.41 mmol) was dissolved in dry THF (50 mL), and the solution was cooled to -78 °C. KHMDS solution (5.11 mL, 5.11 mmol, 1.0 N in THF) was added dropwise. After the addition was completed, the reaction was continued for 1 h. After the addition of a solution of Davis Oxaziridine (1.07 g, 4.09 mmol) in dry THF (10 mL), the reaction was continued for about 4 h. After the completion of the reaction was monitored by TLC, saturated aqueous ammonium chloride solution (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 3). 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 (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 85:15) to give compound 8B (150 mg, 22.9%).

[0520] LC-MS (ESI): m / z = 193.1 [M+H] + .

[0521] Third Step: Compound 8B (150 mg, 0.781 mmol) was dissolved in a mixture of tetrahydrofuran (2 mL), methanol (2 mL) and water (1 mL), and lithium hydroxide monohydrate (131 mg, 3.12 mmol) was added at room temperature. After the addition was completed, the reaction was continued. After the completion of the reaction was monitored by TLC, the mixture was directly concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (dichloromethane:methanol (v:v) = 100:0 ~ 66:34) to give compound 8C (110 mg, 79.1%).

[0522] LC-MS (ESI): m / z = 177.1 [M-H] - .

[0523] Fourth Step: Compound 8C (110 mg, 0.618 mmol), mesylate of compound 1F (329 mg, 0.618 mmol), HATU (470 mg, 1.24 mmol), HOBT (167 mg, 1.24 mmol) and DIPEA (399 mg, 3.09 mmol) were dissolved in dry N,N-dimethylformamide (20 mL) and the mixture was stirred at room temperature overnight. After the completion of the reaction was monitored by TLC, saturated aqueous ammonium chloride solution (50 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 5). 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 (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 20:80) to give a mixture of compound 8-1 and 8-2 (170 mg, 46.2%). Compound 8-1 (43 mg, 11.7%) and compound 8-2 (30 mg, 8.16%) were obtained by separation and purification by preparative HPLC.

[0524] Preparation HPLC separation and purification method: 1. Instrument: waters 2767 preparative liquid; column: SunFire@Prep C18(19mmx250mm) 2. The sample was filtered with 0.45 μm filter head, and prepared into sample solution. 3. Preparation chromatographic conditions: a. mobile phase A, B composition: mobile phase A: acetonitrile; mobile phase B: water (containing 0.5% ammonium acetate); b. gradient elution, mobile phase A content from 5%-50%; c. flow rate 12 mL / min; d. elution time 30 min.

[0525] Compound 8-1:

[0526] 1 H NMR (400 MHz, Chloroform-d) δ 7.76 (s, 1H), 7.70 (d, 1H), 7.58-7.50 (m, 1H), 5.66-5.51 (m, 2H), 5.31-5.09 (m, 3H), 4.21 (d, 1H), 3.29-3.12 (m, 2H), 2.99-2.59 (m, 5H), 2.43 (s, 3H), 2.39-2.28 (m, 2H), 1.92-1.83 (m, 2H), 1.02 (t, 3H).

[0527] LC-MS (ESI): m / z = 596.2 [M+H] + .

[0528] Compound 8-2:

[0529] 1 H NMR (400 MHz, Chloroform-d) δ 7.55 (d, 1H), 7.48 (s, 1H), 7.39 (s, 1H), 5.66 (d, 1H), 5.31 (d, 1H), 5.15 (d, 1H), 4.67 (dd, 2H), 4.42 (d, 1H), 3.21 (d, 1H), 3.10 (d, 1H), 3.00-2.65 (m, 5H), 2.44-2.28 (m, 4H), 2.07-1.98 (m, 1H), 1.84-1.72 (m, 2H), 1.02 (t, 3H).

[0530] LC-MS (ESI): m / z = 596.2 [M+H] + .

[0531] Example 9

[0532] First step: Compound 9A (8.36 g, 50.0 mmol) was dissolved in super dry DMF (80 mL) under nitrogen protection, and then cooled to about 0 °C. Then NaH (2.4 g, 60 mmol, 60%) was slowly added. After the addition was completed, the reaction was continued at this temperature for 1 h. Then 3- iodyloxetane (18.4 g, 100 mmol) was added, and then the reaction was stirred at room temperature for 18 h. Then water was added to the reaction solution, and then extracted twice with ethyl acetate. The organic phase was combined and washed with water and saturated brine successively. Then the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by column chromatography to give the target compound 9B (9.6 g, 85.94%).

[0533] 1 H NMR (400 MHz, CDCl3) δ 7.86-7.83 (m, 1H), 7.76-7.74 (m, 1H), 7.43-7.39 (m, 1H), 7.32-7.28 (m, 1H), 5.20-5.16 (m, 2H), 5.08-5.01 (m, 1H), 4.76-4.73 (m, 2H).

[0534] Second step: Compound 9B (9.6 g, 42.99 mmol) was dissolved in methanol (144 mL), and then sodium tungstate (12.825 g, 42.99 mmol) was added. Then hydrogen peroxide (26 mL) was slowly added under stirring, and then the reaction was stirred at room temperature overnight. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was filtered, and then the filter cake was washed with ethyl acetate. The aqueous phase was separated, and then the organic phase was washed with sodium thiosulfate aqueous solution and saturated brine successively. Then the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by column chromatography to give the target compound 9C (2.2 g, 20.05%).

[0535] LC-MS (ESI): m / z = 256.0 [M+H] + .

[0536] Third step: Methyl 2-(3-oxocyclobutyl)acetate (14.2 g, 100 mmol) was dissolved in methanol (150 mL), and then trimethyl orthoformate (40.95 g, 400 mmol) and p-toluenesulfonic acid monohydrate (1.90 g, 10 mmol) were added successively. Then the reaction was stirred at room temperature overnight. The reaction was monitored by TLC (DNP coloration). After the reaction was completed, excess saturated sodium carbonate aqueous solution was added to the reaction solution to make it alkaline. Then the reaction solution was extracted twice with ethyl acetate. The organic phase was combined and washed with water and saturated brine successively. Then the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by column chromatography to give the target compound 9D (16.1 g).

[0537] 1H NMR (400 MHz, CDC13) δ 3.66 (s, 3H), 3.15 (s, 3H), 3.13 (s, 3H), 2.50-2.37 (m, 5H), 1.84-1.76 (m, 2H).

[0538] Fourth step: 9D (9.40 g, 50.0 mmol) was weighed in anhydrous THF (80 mL), and the reaction solution was cooled to about -70°C in a dry ice-ethanol bath under nitrogen protection, then KHMDS THF solution (70 mL, 70 mmol, 1 mol / L) was slowly added, and after the addition was completed, the temperature was kept for 1 h of stirring, then 3-phenyl-2-phenylsulfonyl-1,2-oxaziridine (18.28 g, 70 mmol) was dissolved in anhydrous THF (80 mL), and the solution was slowly added to the above reaction solution, and after the addition was completed, the reaction was continued for 2 h at the same temperature, and TLC was used to monitor the reaction (potassium permanganate coloration). After the reaction was completed, saturated ammonium chloride aqueous solution and water were added to the reaction solution, then ethyl acetate was added for extraction twice, the organic phase was combined, and then washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified by column chromatography to obtain the target compound 9E (5.10 g, 49.95%).

[0539] 1 H NMR (400 MHz, CDC13) δ 4.16-4.15 (m, 1H), 3.77 (s, 3H), 3.16 (s, 3H), 3.14 (s, 3H), 2.47-2.37 (m, 1H), 2.30-2.04 (m, 4H).

[0540] Fifth step: 9E (5.10 g, 24.97 mmol) was weighed in DMF (51 mL), then tert-butyl diphenylchlorosilane (13.726 g, 49.94 mmol), imidazole (8.50 g, 124.85 mmol) and DMAP (3.05 g, 24.97 mmol) were added in sequence, and after the addition was completed, the reaction was stirred at room temperature for 2 h, and TLC was used to monitor the reaction, then water was added to the reaction solution, and ethyl acetate was added for extraction twice, the organic phase was combined, and then washed with water, saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the target compound 9F (13.1 g).

[0541] 1H NMR (400 MHz, CDC13) δ 7.65-7.62 (m, 4H), 7.44-7.33 (m, 6H), 4.19-4.18 (m, 1H), 3.38 (s, 3H), 3.13 (s, 3H), 3.06 (s, 3H), 2.44-2.37 (m, 1H), 2.19-2.00 (m, 4H), 1.10 (m, 9H).

[0542] Step 6: Take 9F (13.10 g, crude) in acetonitrile (130 mL), then add p-toluenesulfonic acid monohydrate (5.63 g, 29.60 mmol), after the addition is complete, stir the reaction at room temperature for 2 h, after the reaction is complete, directly concentrate, then purify by column chromatography to obtain the target compound 9G (7.50 g).

[0543] 1 H NMR (400 MHz, CDC13) δ 7.65-7.62 (m, 4H), 7.44-7.33 (m, 6H), 4.19-4.18 (m, 1H), 3.38 (s, 3H), 3.13 (s, 3H), 3.06 (s, 3H), 2.44-2.37 (m, 1H), 2.19-2.00 (m, 4H), 1.10 (m, 9H).

[0544] Step 7: Take compound 9G (2.40 g, 6.05 mmol) and compound 9C (1.542 g, 6.05 mmol) in super dry THF (30 mL), replace with nitrogen protection, then cool to 0°C, slowly add LHMDS (7.3 mL, 7.3 mmol, 1.0 mol / L) in tetrahydrofuran solution to the reaction solution, after the addition is complete, naturally warm to room temperature overnight, after the reaction is complete, directly concentrate and dry, then purify by column chromatography to obtain the target compound 9H (1.90 g, 71.97%).

[0545] 1 H NMR (400 MHz, CDC13) δ 7.65-7.62 (m, 4H), 7.44-7.33 (m, 6H), 4.19-4.18 (m, 1H), 3.38 (s, 3H), 3.13 (s, 3H), 3.06 (s, 3H), 2.44-2.37 (m, 1H), 2.19-2.00 (m, 4H), 1.10 (m, 9H).

[0546] Eighth step: Take compound 9H (1.90 g, 4.35 mmol) in methanol, tetrahydrofuran and water (MeOH:THF:H2O = 2:2:1) mixed solvent 50 mL, then add lithium hydroxide (417 mg, 17.4 mmool), react at room temperature overnight, after the reaction is completed, the solvent is concentrated, then water is added, 3N HC1 is used to adjust pH = 2-3, ethyl acetate is extracted twice, the organic phase is combined, anhydrous sodium sulfate is used to dry the organic phase, the concentrated crude product is obtained, and the target compound 9I (1.30 g, 70.65%) is obtained after column chromatography purification of the crude product.

[0547] LC-MS (ESI): m / z = 421.0 [M-1] + .

[0548] Ninth step: Take compound 9I (522 mg, 1.24 mmol), exatecan methanesulfonate (660 mg, 1.24 mg) in DMF (10 mL) in turn, then add HOBT (202 mg, 1.49 mmol), DIPEA (640 mg, 4.96 mmol) and HATU (566 mg, 1.49 mmol) in turn, after the addition is completed, stir at room temperature overnight, monitor the reaction by TLC, after the reaction is completed, pour the reaction liquid into water, extract with ethyl acetate, dry with anhydrous sodium sulfate, filter, concentrate to obtain the crude product, and the target compound 9J crude product (620 mg, 59.56%) is obtained after column chromatography purification of the crude product. The compound is directly used in the next step reaction.

[0549] Tenth step: Take compound crude product 9J (620 mg, 0.738 mmol) in tetrahydrofuran (4 mL), then add hydrogen fluoride triethylamine solution (5 mL), stir at room temperature for 24 h, TLC spot half of the reaction, add hydrogen fluoride triethylamine solution (5 mL) again, stir for 24 h again, TLC spot half of the remaining small amount of raw material, add water to the reaction liquid, extract twice with ethyl acetate, then wash the organic phase with saturated brine, add DIPEA to the ethyl acetate phase to make it basic, then wash the organic phase with water twice again, dry with anhydrous sodium sulfate, filter, concentrate to obtain the crude product, and the target compound 9-1 (76 mg, 17.16%) and the target compound 9-2 (84 mg, 18.96%) are obtained after reverse phase preparation purification of the crude product. Purification method: instrument: Shimadzu LC-20AP; column: C18; mobile phase: A is water; B is acetonitrile; gradient: B changes from 33% to 63% in 15 minutes; flow rate: 25 mL / min; column temperature: room temperature; detection wavelength: 210 nm and 254 nm; compound 9-1 retention time tR=1.457; compound 9-2 retention time tR=1.524.

[0550] Compound 9-1: LC-MS (ESI): m / z = 602.3 [M+H]+ .

[0551] 1 H NMR (400 MHz, DMSO-d6) δ 8.41-8.39 (m, 1H), 7.79-7.76 (m, 1H), 7.30 (s, 1H), 6.48 (s, 1H), 5.63-5.62 (m, 1H), 5.58-5.53 (m, 1H), 5.46-5.38 (m, 2H), 5.20-5.11 (m, 2H), 5.03-4.95 (m, 4H), 3.96-3.93 (m, 1H), 3.22-3.11 (m, 2H), 2.79-2.68 (m, 1H), 2.58-2.54 (m, 4H), 2.39 (s, 3H), 2.19-2.12 (m, 2H), 1.93-1.79 (m, 2H), 0.89-0.86 (m, 3H).

[0552] Compound 9-2: LC-MS (ESI): m / z = 602.3 [M+H] + .

[0553] 1 H NMR (400 MHz, DMSO-d6) δ 8.38-8.36 (m, 1H), 7.78-7.75 (m, 1H), 7.31 (s, 1H), 6.49 (s, 1H), 5.60-5.59 (m, 1H), 5.56-5.51 (m, 1H), 5.46-5.38 (m, 2H), 5.23-5.13 (m, 2H), 5.02-4.94 (m, 4H), 4.00-3.98 (m, 1H), 3.21-3.10 (m, 2H), 2.79-2.66 (m, 1H), 2.63-2.45 (m, 4H), 2.39 (s, 3H), 2.19-2.09 (m, 2H), 1.92-1.81 (m, 2H), 0.89-0.86 (m, 3H).

[0554] Example 10:

[0555] Compound 10-1 (LC-MS (ESI): m / z = 600.2 [M+H] + ) and 10-2 (LC-MS (ESI): m / z = 600.2 [M+H] + ) were synthesized according to Example 9.

[0556] Example 11:

[0557] First step: Compound 1E (58.2 mg, 0.409 mmol), trifluoroacetate salt of compound 11A (150 mg, 0.273 mmol, preparation method refer to WO 2022 / 078260), HATU (208 mg, 0.546 mmol), HOBT (73.8 mg, 0.546 mmol) and DIPEA (176 mg, 1.36 mmol) were dissolved in dry N,N-dimethylformamide (8 mL) and reacted at room temperature overnight. After monitoring the completion of the reaction by TLC, saturated aqueous ammonium chloride solution (50 mL) was added, and ethyl acetate was extracted (20 mL x 5), the combined organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 20:80) to obtain a mixture of compounds 11-1 and 11-2 (117 mg, 60.7%), which were separated by SFC to obtain two isomers, compound 11-1 (40 mg, retention time 1.582 min, 20.7%) and compound 11-2 (47 mg, retention time 1.662 min, 22.4%).

[0558] Analytical HPLC conditions: Instrument: CAS-05-LCMS-P; Column: C18. Mobile phase A, B composition: A 0.0375% trifluoroacetic acid aqueous solution; B 0.0185% trifluoroacetic acid acetonitrile solution, gradient elution, B from 5% to 95% in 4 min, flow rate 1 mL / min.

[0559] Preparative HPLC separation conditions: Instrument: CAS-05-Semi-prep K; Column: SunFire® Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A is 0.1% trifluoroacetic acid aqueous solution; B is acetonitrile, gradient elution, B from 45% to 60% in 18 min, flow rate 30 mL / min. Elution time 25 min.

[0560] Compound 11-1:

[0561] 1H NMR (400 MHz, DMSO-d6) δ 8.43 (d, 1H), 7.77 (dd, 1H), 7.31 (s, 1H), 6.48 (s, 1H), 5.61 (d, 1H), 5.58 - 5.51 (m, 1H), 5.50 - 5.36 (m, 2H), 5.19 - 5.09 (m, 2H), 4.74 - 4.67 (m, 2H), 3.98 - 3.90 (m, 1H), 3.22 - 3.07 (m, 2H), 2.74 - 2.67 (m, 3H), 2.67 - 2.59 (m, 2H), 2.42 - 2.35 (m, 3H), 2.20 - 2.09 (m, 2H), 1.97 - 1.80 (m, 2H), 0.88 (t, 3H).

[0562] LC-MS (ESI): m / z = 560.3 [M+H] + .

[0563] Compound 11-2:

[0564] 1 H NMR (400 MHz, DMSO-d6) δ 8.43 (d, 1H), 7.77 (dd, 1H), 7.31 (s, 1H), 6.48 (s, 1H), 5.61 (d, 1H), 5.58 - 5.51 (m, 1H), 5.50 - 5.36 (m, 2H), 5.19 - 5.09 (m, 2H), 4.74 - 4.67 (m, 2H), 3.98 - 3.90 (m, 1H), 3.22 - 3.07 (m, 2H), 2.74 - 2.67 (m, 3H), 2.67 - 2.59 (m, 2H), 2.42 - 2.35 (m, 3H), 2.20 - 2.09 (m, 2H), 1.97 - 1.80 (m, 2H), 0.88 (t, 3H).

[0565] LC-MS (ESI): m / z = 560.3 [M+H] + .

[0566] Example 12:

[0567] First step: Compound 12A (25.0 g, 131 mmol) was dissolved in dichloromethane (500 mL) and cooled to 0 °C. Imidazole (13.4 g, 197 mmol), triphenylphosphine (41.3 g, 158 mmol) and iodine (46.7 g, 184 mmol) were added successively. The reaction was stirred at room temperature. TLC monitoring showed that the reaction was completed. Saturated aqueous ammonium chloride solution (500 mL) was added, and dichloromethane (100 mL x 5) was used for extraction. 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 (v:v) = 100:0 ~ 90:10) to obtain compound 12B (30.0 g, 76.1%).

[0568] Second step: Compound 12B (30.0 g, 100 mmol), triphenylphosphine (26.2 g, 100 mmol) and DIPEA (80 mL) were added to a round-bottom flask and heated to 80 °C overnight to generate a large amount of white solid. After the reaction was completed by TLC monitoring, it was cooled to room temperature, filtered, and the filter cake was washed with n-hexane. The filter cake was collected and dried to obtain compound 12C (45.0 g, 80.1%), which was directly used in the next step.

[0569] LC-MS (ESI): m / z = 436.3 [M+H] + .

[0570] Third step: Compound 12C (27.6 g, 49.1 mmol) was dissolved in dry THF (300 mL) under nitrogen protection, and potassium tert-butoxide (5.05 g, 45.0 mmol) was added at 0 °C. After stirring for 1 h, 1-Boc-3-azetidinone (7.00 g, 40.9 mmol) in dry THF (50 mL) was added, and the reaction was allowed to warm to room temperature overnight. After the reaction was completed by TLC monitoring, saturated aqueous ammonium chloride solution (200 mL) was added, and ethyl acetate (100 mL x 5) was used for extraction. 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 (v:v) = 100:0 ~ 90:10) to obtain compound 12D (4.10 g, 30.6%).

[0571] LC-MS (ESI): m / z = 328.2 [M+H] + .

[0572] Fourth step: Compound 12D (2.00 g, 6.11 mmol) was dissolved in tetrahydrofuran (50 mL), TBAF (12.2 mL, 12.2 mmol, 1 N in THF) was added at room temperature, and the reaction was continued. After the completion of the reaction was monitored by TLC, it was directly concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 50:50) to obtain compound 12E (1.22 g, 93.8%).

[0573] LC-MS (ESI): m / z = 214.2 [M+H] + .

[0574] Fifth step: Compound 12E (700 mg, 3.28 mmol) was dissolved in dichloromethane (50 mL), DMP (2.78 g, 6.56 mmol) was added at room temperature, and the reaction was continued. After the completion of the reaction was monitored by TLC, the solid was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 65:35) to obtain compound 12F (500 mg, 72.2%).

[0575] LC-MS (ESI): m / z = 212.2 [M+H] + .

[0576] Sixth step: 12F (500 mg, 2.37 mmol) was dissolved in a mixture solvent of tert-butyl alcohol (15 mL), water (15 mL) and 2-methyl-2-butene (7.5 mL), and sodium phosphate dibasic dihydrate (1.85 g, 11.8 mmol), sodium chlorite (429 mg, 4.74 mmol) were sequentially added at 0°C, and the reaction was stirred at room temperature. After the completion of the reaction was monitored by TLC, saturated aqueous ammonium chloride solution (50 mL) was added, dichloromethane (20 mL x 5) was extracted, the combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 100:0 ~ 75:15) to obtain compound 12G (270 mg, 50.2%).

[0577] LC-MS (ESI): m / z = 226.1 [M-H] - .

[0578] Seventh step: Compound 12G (300 mg, 1.32 mmol), methanesulfonate of compound 1F (701 mg, 1.32 mmol), HATU (1.00 g, 2.64 mmol), HOBT (357 mg, 2.64 mmol) and DIPEA (681 mg, 5.28 mmol) were dissolved in dry N,N-dimethylformamide (20 mL) and reacted at room temperature overnight. After monitoring the completion of the reaction by TLC, saturated aqueous ammonium chloride solution (50 mL) was added, and ethyl acetate (20 mL x 5) was extracted. The organic phase was combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v:v) = 100:0 ~ 20:80) to obtain compound 12H (500 mg, 47.0%),

[0579] LC-MS (ESI): m / z = 645.3 [M+H] + .

[0580] Eighth step: Compound 12H (200 mg, 0.310 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added and stirred at room temperature. After monitoring the disappearance of the raw material by TLC, it was directly concentrated under reduced pressure and redissolved in dichloromethane and concentrated under reduced pressure three times to remove excess trifluoroacetic acid. The obtained oily 12I (220 mg) was used directly in the next step without purification.

[0581] LC-MS (ESI): m / z = 545.3 [M+H] + .

[0582] Ninth step: Compound 12I (220 mg) obtained in the previous step, 2-bromoethanol (77.5 mg, 0.620 mmol), DIPEA (200 mg, 1.55 mmol), and DMAP (37.9 mg, 0.310 mmol) were all dissolved in 1,2-dichloroethane (20 mL) and heated to 50°C for reaction. After monitoring the disappearance of the raw material by LCMS, it was directly concentrated under reduced pressure, and the obtained residue was purified by preparative liquid chromatography to obtain compound 12 (4 mg, 2.2% yield in two steps).

[0583] Preparative HPLC separation conditions: Instrument: CAS-05-Prep-HPLC-S; Column: SunFire@Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A 0.1% trifluoroacetic acid aqueous solution; B acetonitrile, gradient elution, B from 45% to 60% in 18 min, flow rate 30 mL / min. Elution time 25 min.

[0584] 1H NMR (400 MHz, Methanol-d4) δ 7.68-7.55 (m, 2H), 7.42 (d, 1H), 6.31 (d, 1H), 5.94 (s, 1H), 5.77-5.65 (m, 2H), 5.56 (d, 1H), 5.34 (d, 1H), 5.19 (d, 1H), 4.97 (d, 1H), 3.97 (s, 2H), 3.88-3.80 (m, 2H), 3.35-3.25 (overlapped, 2H), 3.24-3.17 (m, 2H), 2.47-2.34 (m, 5H), 2.28-2.17 (m, 2H), 2.00-1.90 (m, 2H), 1.00 (t, 3H).

[0585] LC-MS (ESI): m / z = 589.2 [M+H] + .

[0586] Example 13:

[0587] First Step: Potassium tert-butoxide (14.6 g, 131.55 mmol) was added to a reaction flask, mixed with DMF (100 ml), and cooled to -45 °C. Compound 13A (20 g, 82.22 mmol) was dissolved in DMF (100 ml) and added to the above system. Then 2-(difluoromethyl)sulfonyl)pyridine (15.88 g, 82.22 mmol) was dissolved in DMF (100 ml) and added to the above system. The system was slowly warmed to room temperature and reacted for 16 hours. Then saturated ammonium chloride solution (200 ml) and 3 mol / L hydrochloric acid solution (80 ml) were added, and stirred at room temperature for 3 hours. Water (1.5 L) was added for dilution, and extracted twice with ethyl acetate (300 ml x 2). The organic phase was combined and dried, and purified by silica gel column chromatography to obtain compound 13B (7.0 g, 30.71%).

[0588] 1 H NMR (400 MHz, DMSO-d6) δ 4.48-4.39 (m, 1H), 4.05-3.95 (m, 2H), 3.70-3.63 (m, 3H), 3.10-2.95 (m, 1H), 2.65-2.54 (m, 1H), 1.43-1.32 (m, 9H).

[0589] Second Step: Compound 13B (2.0 g, 7.21 mmol) was added to a reaction flask, and then HCl-1,4-dioxane solution (10 ml) was added and reacted at room temperature for 1 hour. After the reaction was completed by TLC monitoring, it was concentrated under reduced pressure to obtain compound 13C (1.1 g, 71.43%).

[0590] LC-MS (ESI): m / z = 178.2 [M+H] + .

[0591] Third step: Compound 13C (1.1 g, 5.15 mmol) was taken in a reaction flask, dissolved in dichloromethane (30 ml), then acetoxy acetic acid (810 mg, 6.83 mmol) and DIPEA (4.5 ml, 25.75 mmol) were added, finally HATU (2.94 g, 7.72 mmol) was added and the reaction was allowed to proceed for 16 h at room temperature. After completion of the reaction as monitored by TLC, it was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to obtain compound 13D (1.0 g, 70.07%).

[0592] LC-MS (ESI): m / z = 278.1 [M+H] + .

[0593] Fourth step: Compound 13D (1.0 g, 3.61 mmol) was taken in a reaction flask, dissolved in THF:MeOH:H2O = 1:1:1 (15 ml), then lithium hydroxide (346 mg, 14.44 mmol) was added and the reaction was allowed to proceed for 16 h at room temperature. After completion of the reaction as monitored by TLC, it was adjusted to pH 2-3 with concentrated hydrochloric acid and purified by reverse phase column to obtain compound 13E (0.3 g, 37.59%).

[0594] LC-MS (ESI): m / z = 222.0 [M+H] + .

[0595] Fifth step: Exetecan mesylate (0.3 g, 0.56 mmol), compound 13E (250 mg, 1.12 mmol), HOBT (114 mg, 0.84 mmol) and DIPEA (0.5 ml, 2.80 mmol) were taken in a reaction flask, dissolved in DMF (10 ml), then EDCI (160 mg, 0.84 mmol) was added and the reaction was allowed to proceed for 16 h at room temperature. After completion of the reaction as monitored by TLC, it was purified by reverse phase column chromatography to obtain compound 13 (120 mg, 33.56%).

[0596] 1H NMR (400 MHz, DMSO-d6) δ 8.80-8.60 (m, 1H), 7.83-7.76 (m, 1H), 7.31 (s, 1H), 6.51-6.48 (m, 1H), 5.58-5.46 (m, 1H), 5.43 (s, 2H), 5.33-5.00 (m, 2H), 4.98-4.77 (m, 1H), 4.66-4.57 (m, 1H), 4.27-4.01 (m, 4H), 3.22-3.12 (m, 2H), 3.00-2.54 (m, 2H), 2.41 (s, 3H), 2.22-2.08 (m, 2H), 1.94-1.79 (m, 2H), 0.92-0.84 (m, 3H).

[0597] LC-MS (ESI): m / z = 639.2 [M+H] + .

[0598] Example 14:

[0599] First Step: Compound 11A (100 mg, 0.11 mmol), compound 5D (41 mg, 0.22 mmol), HOBT (22 mg, 0.16 mmol) and DIPEA (0.2 ml, 1.10 mmol) were taken in a reaction vial, dissolved in DMF (5 ml), then EDCI (31 mg, 0.16 mmol) was added and allowed to react at room temperature for 16 h. After completion of the reaction as monitored by TLC, it was purified by reverse phase column to get compound 14 (12 mg, yield: 17.39%).

[0600] 1 H NMR (400 MHz, DMSO-d6) δ 8.65-8.52 (m, 1H), 7.82-7.77 (m, 1H), 7.33 (s, 1H), 6.50 (s, 1H), 5.57-5.46 (m, 1H), 5.43 (s, 2H), 5.31-5.12 (m, 2H), 5.09-5.00 (m, 2H), 4.55-4.44 (m, 1H), 4.16-4.09 (m, 2H), 4.08-3.98 (m, 2H), 3.21-3.13 (m, 2H), 2.94-2.82 (m, 1H), 2.72-2.53 (m, 2H), 2.41 (s, 3H), 2.17-2.04 (m, 2H), 1.93-1.79 (m, 2H), 0.91-0.83 (m, 3H).

[0601] LC-MS (ESI): m / z = 603.0 [M+H] + .

[0602] Example 15:

[0603] First Step: Compound 9A (900 mg, 5.38 mmol) was taken in a reaction flask, dissolved in DMF (20 ml), NaH (226 mg, 5.65 mmol) was added, stirred for 30 min at room temperature, then compound 15A (1.5 g, 4.87 mmol) was added, reacted for 16 h at room temperature. After completion of the reaction monitored by TLC, diluted with water (200 ml), extracted with ethyl acetate twice (100 ml x 2), combined the organic layers and dried, concentrated under reduced pressure, the residue obtained was purified by silica gel column chromatography to get compound 15B (1.2 g, 62.50 %).

[0604] LC-MS (ESI): m / z = 395.1 [M+H] + .

[0605] Second Step: Compound 15B (1.2 g, 3.04 mmol) and sodium tungstate (893 mg, 3.04 mmol) were taken in a reaction flask, dissolved in MeOH (30 ml), then H2O2 (3.6 ml) was added slowly, reacted for 16 h at room temperature. After completion of the reaction monitored by TLC, concentrated under reduced pressure, the residue obtained was purified by silica gel column chromatography to get compound 15C (960 mg, 74.07 %).

[0606] LC-MS (ESI): m / z = 327.0 [M-100+H] + .

[0607] Third Step: Compound 15C (960 mg, 2.25 mmol) and 3-oxetanone (1.62 g, 22.50 mmol) were taken in a reaction flask, dissolved in THF (100 ml), then LiHMDS (5.63 ml, 5.63 mmol) was added slowly, reacted for 16 h at room temperature. After completion of the reaction monitored by TLC, added saturated ammonium chloride solution (10 ml), concentrated under reduced pressure, the residue obtained was purified by silica gel column chromatography to get compound 15D (185 mg, 29.04 %).

[0608] LC-MS (ESI): m / z = 184.2 [M-100+H] + .

[0609] Fourth Step: Compound 15D (185 mg, 0.65 mmol) was taken in a reaction flask, dissolved in DCM (10 ml), then TFA (1 ml) was added, reacted for 16 h at room temperature. After completion of the reaction monitored by TLC, concentrated under reduced pressure to get compound 15E (120 mg, 100.84 %).

[0610] LC-MS (ESI): m / z = 184.1 [M+H] + .

[0611] Fifth step: Compound 15E (120 mg, 0.65 mmol), 2-((tetrahydro-2H-pyran-2- yl)oxy)acetic acid (208 mg, 1.30 mmol, synthesized as per patent EP3656782B1) and DIPEA (0.57 ml, 3.25 mg) were taken in a reaction vial, dissolved in DCM (20 ml), followed by addition of HATU (369 mg, 0.97 mmol) and allowed to react for 16 h at room temperature. The completion of the reaction was monitored by TLC, after which it was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to obtain compound 15F (100 mg, 47.39%).

[0612] LC-MS (ESI): m / z = 242.0 [M-84+H] + .

[0613] Sixth step: Compound 15F (100 mg, 0.31 mmol) was taken in a reaction vial, dissolved in THF:MeOH:H2O = 1:1:1 (6 ml), followed by addition of lithium hydroxide (9 mg, 0.37 mmol) and allowed to react for 16 h at room temperature. The completion of the reaction was monitored by TLC, after which it was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to obtain compound 15G (30 mg, 31.09%).

[0614] LC-MS (ESI): m / z = 228.1 [M-84+H] + .

[0615] Seventh step: Exetecan mesylate (35 mg, 0.066 mmol), compound 15G (30 mg, 0.10 mmol), HOBT (14 mg, 0.10 mmol) and DIPEA (0.06 ml, 0.33 mmol) were taken in a reaction vial, dissolved in DCM (5 ml), followed by addition of EDCI (19 mg, 0.10 mmol) and allowed to react for 16 h at room temperature. The completion of the reaction was monitored by TLC, after which it was purified by silica gel column to obtain compound 15H (20 mg, yield: 41.58%).

[0616] LC-MS (ESI): m / z = 645.2 [M-84+H] + .

[0617] Eighth step: Compound 15H (20 mg, 0.027 mmol) was added to the reaction bottle, dissolved with MeOH (5 ml), then p-toluenesulfonic acid (4.6 mg, 0.027 mmol) was added, and the reaction was carried out at room temperature for 1 hour. After monitoring the completion of the reaction by TLC, purification was carried out by reverse phase column to obtain compound 15 (1.2 mg, yield: 7.58%).

[0618] 1 H NMR (400 MHz, DMSO-d6) δ 8.76-8.65 (m, 1H), 7.84-7.78 (s, 1H), 7.33 (s, 1H), 6.51-6.49 (s, 1H), 5.98-5.90 (m, 1H), 5.47-5.42 (m, 2H), 5.39-5.30 (m, 2H), 5.24-5.20 (m, 1H), 5.13-5.09 (m, 1H), 4.99-4.90 (m, 1H), 4.45-4.36 (m, 1H), 4.35-4.31 (m, 1H), 4.23-4.03 (m, 3H), 3.46-3.43 (m, 2H), 2.41 (s, 3H), 2.03-1.95 (m, 4H), 1.90-1.84 (m, 2H), 1.51-1.41 (m, 2H), 1.08-1.03 (m, 3H).

[0619] LC-MS (ESI): m / z = 645.0 [M+H] + .

[0620] Example 16:

[0621] First step: Compound 11A trifluoroacetate (132 mg, 0.24 mmol) and compound 9I (100 mg, 0.24 mmol) were weighed into DMF (5 mL) in turn, then HOBT (39 mg, 0.29 mmol), DIPEA (93 mg, 0.72 mmol) and HATU (110 mg, 0.29 mmol) were added in turn, and the reaction was stirred at room temperature overnight after completion. TLC monitoring of the reaction, the reaction was completed, the reaction liquid was poured into water and stirred for 10 min, filtered and dried to obtain the crude product of the target compound 16A (150 mg, 74.62%), which was directly used in the next step reaction.

[0622] Second step: Take compound crude product 16A (150 mg, 0.179 mmol) in tetrahydrofuran (2 mL), then add hydrogen fluoride triethylamine solution (2 mL), after adding, stir at room temperature for 24 h, add hydrogen fluoride triethylamine solution (2 mL) again, stir for 24 h again, add water to the reaction solution, extract twice with ethyl acetate, then wash the organic phase with saturated brine, add DIPEA to the ethyl acetate phase to make it basic, then wash the organic phase with water twice, dry over anhydrous sodium sulfate, filter, concentrate to obtain the crude product, purify the crude product by reverse phase preparative purification to obtain target compound 16-1 (13 mg, 9.03%, two-step yield) and target compound 16-2 (16 mg, 11.11%, two-step yield).

[0623] Purification method: First purification: instrument: CAS-05-Semi-prep K; column: C18; mobile phase: A is water; B is acetonitrile; flow rate: 25 mL / min; column temperature: room temperature; wavelength: 220 and 254 nanometers; secondary purification of P2: instrument: CAS-05-Semi-prep K; column: C18; mobile phase: A is water; B is acetonitrile; flow rate: 25 mL / min; column temperature: room temperature; wavelength: 220 and 254 nanometers; compound 16-1 retention time t R = 2.208 min; compound 16-2 retention time t R = 2.312 min.

[0624] Compound 16-1: LC-MS (ESI): m / z = 602.2 [M+H] + .

[0625] 1 H NMR (400 MHz, DMSO-d6) δ 8.47-8.44 (m, 1H), 7.79-7.76 (m, 1H), 7.30 (s, 1H), 6.49 (s, 1H), 5.65-5.63 (m, 1H), 5.57-5.52 (m, 1H), 5.43 (s, 2H), 5.20-5.11 (m, 2H), 5.03-4.95 (m, 4H), 3.96-3.94 (m, 1H), 3.22-3.11 (m, 2H), 2.79-2.73 (m, 1H), 2.60-2.54 (m, 4H), 2.37 (s, 3H), 2.15-2.13 (m, 2H), 1.92-1.79 (m, 2H), 0.89-0.85 (m, 3H).

[0626] Compound 16-2: LC-MS (ESI): m / z = 602.2 [M+H] + .

[0627] 1 H NMR (400 MHz, DMSO-d6) δ 8.43-8.41 (m, 1H), 7.80-7.77 (m, 1H), 7.31 (s, 1H), 6.49 (s, 1H), 5.67-5.66 (m, 1H), 5.56-5.53 (m, 1H), 5.42 (m, 2H), 5.23-5.14 (m, 2H), 5.02-4.99 (m, 4H), 4.01-3.98 (m, 1H), 3.18-3.12 (m, 2H), 2.73-2.66 (m, 1H), 2.63-2.45 (m, 4H), 2.39 (s, 3H), 2.19-2.08 (m, 2H), 1.92-1.81 (m, 2H), 0.89-0.86 (m, 3H).

[0628] 2. Synthesis of linker conjugated toxins

[0629] Example L-1:

[0630] First step: Compound 1D (4.00 g, 25.6 mmol) was dissolved in dry DMF (100 mL), and cooled to 0 °C, then compound L-1A (28.3 g, 76.8 mmol) and hydrochloric acid dioxane solution (3.8 mL, 15.4 mmol, 4N) were added successively, and then the temperature was allowed to rise to room temperature and the reaction was allowed to proceed overnight. After the disappearance of the starting material was monitored by TLC, saturated aqueous sodium bicarbonate solution (100 mL) and water (400 mL) were added, and extracted with ethyl acetate (200 mL x 5), and the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: acetone (v:v) = 100:0 ~ 70:30) to give compound L-1B (4.50 g, 37.8%).

[0631] LC-MS (ESI): m / z = 465.2 [M+H] + .

[0632] Second step: Compound L-1B (4.20 g, 9.04 mmol) was dissolved in dry DMF (30 mL), and triethylamine (1.10 g, 10.9 mmol) was added dropwise, and the reaction was allowed to proceed overnight after the addition was completed. After the completion of the reaction was monitored by LCMS, it was directly purified by C-18 silica reverse phase chromatography (water: acetonitrile (v:v) = 98:2 ~ 50:50) to give compound L-1C (1.48 g, 67.6%).

[0633] LC-MS (ESI): m / z = 243.1 [M+H] + .

[0634] Step 3: Compound L-1C (1.48 g, 6.11 mmol) was dissolved in DMF (30 mL), compound L-1D (3.06 g, 6.11 mmol), pyBOP (3.81 g, 7.33 mmol) and DIPEA (1.03 g, 7.94 mmol) were added at room temperature, and the reaction was continued. After the reaction was completed by LCMS monitoring, saturated aqueous ammonium chloride solution (100 mL) was added, extracted with ethyl acetate (100 mL x 5), the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane:methanol (v:v) = 100:0 ~ 90:10) to obtain compound L-1E (4.16 g, 93.9%).

[0635] LC-MS (ESI): m / z = 726.2 [M+H] + .

[0636] Step 4: Compound L-1E (8.00 g, 11.0 mmol) was dissolved in a mixed solvent of tetrahydrofuran (30 mL), methanol (30 mL) and water (15 mL), and lithium hydroxide monohydrate (1.39 g, 33.1 mmol) was added at room temperature, and the reaction was continued. After the reaction was completed by TLC monitoring, it was directly concentrated under reduced pressure, and the residue was purified by C-18 silica reverse phase chromatography (water (containing 0.5% ammonium bicarbonate): acetonitrile (v:v) = 98:2 ~ 60:40) to obtain compound L-1F (2.20 g, 40.8%).

[0637] LC-MS (ESI): m / z = 490.2 [M+H] + .

[0638] Step 5: Compound L-1F (2.20 g, 4.49 mmol), compound L-1G (1.52 g, 4.94 mmol) and DIPEA (696 mg, 5.39 mmol) were dissolved in dry N,N-dimethylformamide (30 mL) and reacted at room temperature. After the reaction was completed by LCMS monitoring, it was directly purified by C-18 silica reverse phase chromatography (water: acetonitrile (v:v) = 98:2 ~ 50:50) to obtain compound L-1H (2.60 g, 84.7%).

[0639] LC-MS (ESI): m / z = 681.3 [M-H] - .

[0640] Step 6: Compound L-1H (2.60 g, 3.81 mmol), compound 1F mesylate (2.02 g, 3.81 mmol), HATU (2.17 g, 5.71 mmol), HOBT (772 mg, 5.71 mmol) and DIPEA (1.97 g, 15.2 mmol) were dissolved in dry N,N-dimethylformamide (40 mL) and reacted at room temperature. After the reaction was monitored to be completed by LCMS, the reaction solution was directly sent to preparative HPLC purification to obtain compound L-1-1 (651 mg, retention time 1.713 min, 15.5%) and compound L-1-2 (589 mg, retention time 1.771 min, 14.1%).

[0641] Analytical HPLC conditions: Instrument: Shimadzu LC-20AP; Column: C18. Mobile phase A, B composition: A is 0.0375% trifluoroacetic acid in water; B is 0.0185% trifluoroacetic acid in acetonitrile, gradient elution, B from 5% to 95% in 4 min, flow rate 1 mL / min.

[0642] Preparative HPLC separation conditions: Instrument: Shimadzu LC-20AP; Column: SunFire® Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A is 0.1% trifluoroacetic acid in water; B is acetonitrile, gradient elution, B from 45% to 60% in 18 min, flow rate 12 mL / min. Elution time 25 min.

[0643] Compound L-1-1:

[0644] 1H NMR (400 MHz, DMSO-d6) δ 8.64 (t, 1H), 8.53 (d, 1H), 8.26 (t, 1H), 8.09-8.00 (m, 2H), 7.96 (t, 1H), 7.76 (d, 1H), 7.30 (s, 1H), 7.27-7.13 (m, 5H), 6.98 (s, 2H), 6.48 (s, 1H), 5.63-5.53 (m, 1H), 5.46-5.36 (m, 2H), 5.13 (s, 2H), 4.76-4.67 (m, 3H), 4.57 m 4.41 (m, 2H), 3.97 (d, 1H), 3.78 m 3.53 (m, 6H), 3.35 (t, 2H), 3.27-3.17 (m, 1H), 3.16-3.07 (m, 1H), 3.04-2.96 (m, 1H), 2.81-2.56 (m, 6H), 2.43-2.31 (m, 3H), 2.24-2.13 (m, 2H), 2.08 (t, 2H), 1.93-1.79 (m, 2H), 1.52-1.40 (m, 4H), 1.25-1.11 (m, 2H), 0.88 (t, 3H).

[0645] LC-MS (ESI): m / z = 1100.6 [M+H] + .

[0646] Compound L-1-2:

[0647] 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (t, 1H), 8.58 (d, 1H), 8.25 (t, 1H), 8.10-8.01 (m, 2H), 7.97 (t, 1H), 7.75 (d, 1H), 7.32 (s, 1H), 7.24 (d, 2H), 7.24 m 7.13 (m, 3H), 6.97 (s, 2H), 5.59-5.52 (m, 1H), 5.47-5.38 (m, 2H), 5.21 (s, 2H), 4.86-4.66 (m, 3H), 4.60-4.44 (m, 2H), 4.06-4.00 (m, 1H), 3.78-3.59 (m, 6H), 3.36 (t, 2H), 3.21-3.00 (m, 3H), 2.84-2.72 (m, 1H), 2.71-2.57 (m, 5H), 2.37 (s, 3H), 2.21-2.06 (m, 4H), 1.93-1.79 (m, 2H), 1.52-1.40 (m, 4H), 1.25-1.11 (m, 2H), 0.88 (t, 3H).

[0648] LC-MS (ESI): m / z = 1100.6 [M+H] + .

[0649] Example L-2:

[0650] First step: Compound 5A (1.0 g, 4.14 mmol) was added to a reaction bottle, dissolved with THF:MeOH:H2O = 3:1:1 (20 ml), then lithium hydroxide (199 mg, 8.28 mmol) was added, and the reaction was carried out at room temperature for 16 hours. After TLC monitoring of the completion of the reaction, the organic solvent was removed by concentration under reduced pressure, the pH was adjusted to about 3 with 3 mol / L hydrochloric acid solution, then extracted with ethyl acetate twice (100 ml x 2), the organic phase was combined and dried, and concentrated under reduced pressure to obtain compound L-2A (900 mg, 95.64%).

[0651] LC-MS (ESI): m / z = 128.1 [M-100+H] + .

[0652] Second step: Compound L-2A (900 mg, 3.96 mmol), exatecan mesylate (1.7 g, 3.17 mmol), HOBT (802 mg, 5.94 mmol) and DIPEA (3.45 ml, 19.8 mmol) were added to a reaction bottle, dissolved with DMF (20 ml), then EDCI (1.14 g, 5.94 mmol) was added, and the reaction was carried out at room temperature for 16 hours. After TLC monitoring of the completion of the reaction, diluted with water (150 ml), then extracted with ethyl acetate three times (100 ml x 3), the organic phase was combined and dried, and concentrated under reduced pressure, the obtained residue was purified by silica gel column chromatography to obtain compound L-2B (1.4 g, 68.62%).

[0653] LC-MS (ESI): m / z = 645.2 [M+H] + .

[0654] Third step: Compound L-2B (1.4 g, 2.17 mmol) was added to a reaction bottle, dissolved with 1,4-dioxane solution (15 ml), then HCl-1,4-dioxane solution (15 ml) was added, and the reaction was carried out at room temperature for 1 hour. After TLC monitoring of the completion of the reaction, concentrated under reduced pressure to obtain compound L-2C (1.2 g, 95.24%).

[0655] LC-MS (ESI): m / z = 545.3 [M+H] + .

[0656] Step 4: To the reaction vial was added compound L-2C (300 mg, 0.51 mmol), L-2D (314 mg, 0.51 mmol, synthesized according to patent WO2019195665A1) and DIPEA (0.44 ml, 2.55 mmol) dissolved in DMF (15 ml) followed by HATU (290 mg, 0.76 mmol) and allowed to react for 16 h at room temperature. After completion of the reaction as monitored by TLC, it was purified by reverse phase column to get compound L-2 (136 mg, 23.32%).

[0657] 1 H NMR (400 MHz, DMSO-d6) d 8.50-8.47 (m, 1H), 8.28-8.22 (m, 1H), 8.12-8.07 (m, 1H), 8.06-8.01 (m, 1H), 8.00-7.95 (m, 1H), 7.82-7.75 (m, 1H), 7.32-7.30 (m, 1H), 7.27-7.14 (m, 5H), 6.98 (s, 2H), 6.50-6.47 (m, 1H), 5.54-5.47 (m, 1H), 5.43 (s, 2H), 5.24-5.18 (m, 2H), 5.07-4.98 (m, 2H), 4.64-4.41 (m, 4H), 4.15-3.93 (m, 4H), 3.77-3.55 (m, 6H), 3.39-3.33 (m, 2H), 3.20-3.12 (m, 2H), 3.09-3.01 (m, 1H), 2.88-2.76 (m, 2H), 2.68-2.55 (m, 1H), 2.40 (s, 3H), 2.15-2.07 (m, 3H), 1.93-1.81 (m, 2H), 1.52-1.41 (m, 4H), 1.26-1.13 (m, 3H), 0.91-0.85 (m, 3H).

[0658] LC-MS (ESI): m / z = 1143.8 [M+H] + .

[0659] Example L-3:

[0660] First step: Compound L-3A (500 mg, 1.16 mmol) was dissolved in dry DMF (5 mL), followed by the addition of compound L-1G (419 mg, 1.36 mmol) and DIPEA (219 mg, 1.70 mmol) and reaction at room temperature. After the disappearance of the starting material was monitored by LCMS, the reaction mixture was directly purified by C-18 silica reverse phase chromatography (water: acetonitrile (with 0.05% trifluoroacetic acid) (v:v) = 98:2 to 50:50) to give compound L-3B (510 mg, 70.9%).

[0661] LC-MS (ESI): m / z = 635.3 [M+H] + .

[0662] Second step: Compound L-3B (500 mg, 0.788 mmol) was dissolved in dry THF (10 mL), followed by the addition of DCC (195 mg, 0.945 mmol) and pentafluorophenol (174 mg, 0.945 mmol). After the completion of the reaction was monitored by LCMS, the reaction mixture was directly evaporated under reduced pressure and the residue was purified by C-18 silica reverse phase chromatography (water: acetonitrile (with 0.05% trifluoroacetic acid) (v:v) = 98:2 to 40:60) to give compound L-3C (520 mg, 82.3%).

[0663] LC-MS (ESI): m / z = 801.3 [M+H] + .

[0664] Third step: Compound L-1F (250 mg, 0.511 mmol), compound L-3C (491 mg, 0.613 mmol) and DIPEA (85.7 mg, 0.664 mmol) were dissolved in dry N,N-dimethylformamide (10 mL) and reaction at room temperature. After the completion of the reaction was monitored by LCMS, the reaction mixture was directly purified by C-18 silica reverse phase chromatography (water: acetonitrile (v:v) = 98:2 to 50:50) to give compound L-3D (216 mg, 84.7%).

[0665] LC-MS (ESI): m / z = 1104.5 [M-H] - .

[0666] Fourth step: Compound L-3D (216 mg, 0.195 mmol), compound 1F mesylate (208 mg, 0.391 mmol) and DMTMM (86.3 mg, 0.293 mmol) were dissolved in dry DMSO (8 mL) and reacted at room temperature. After about 4 hours, the reaction solution was purified by HPLC to obtain compound L-3-1 (83 mg, retention time 2.501 min, 27.9%) and compound L-3-2 (96 mg, retention time 2.546 min, 32.3%).

[0667] Analytical HPLC conditions: Instrument: CAS-05-LCMS-P; Column: C18. Mobile phase A, B composition: A is 0.0375% trifluoroacetic acid in water; B is 0.0185% trifluoroacetic acid in acetonitrile, gradient elution, B from 0% to 80% in 4 min, flow rate 1 mL / min.

[0668] Preparative HPLC separation conditions: Instrument: CAS-05-Semi-prep S; Column: SunFire® Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A is 0.1% trifluoroacetic acid in water; B is acetonitrile, gradient elution, B from 0% to 60% in 18 min, flow rate 30 mL / min. Elution time 25 min.

[0669] Compound L-3-1:

[0670] 1H NMR (400 MHz, DMSO-d6) δ 8.66 (t, 1H), 8.54 (d, 1H), 8.27 (t, 1H), 8.13 (t, 1H), 8.05 (d, 1H), 7.97 (t, 1H), 7.77 (d, 2H), 7.30 (s, 1H), 7.24 (d, 1H), 7.24 - 7.12 (m, 4H), 6.99 (s, 2H), 6.49 (s, 1H), 5.64 - 5.56 (m, 1H), 5.45 - 5.39 (m, 2H), 5.13 (s, 2H), 4.75 - 4.66 (m, 3H), 4.57 - 4.42 (m, 2H), 4.00 - 3.94 (m, 1H), 3.78 - 3.66 (m, 5H), 3.66 - 3.53 (m, 3H), 3.52 - 3.42 (m, 28H), 3.40 - 3.33 (m, 5H), 3.26 - 3.11 (m, 3H), 3.04 - 2.96 (m, 1H), 2.80 - 2.59 (m, 6H), 2.41 - 2.34 (m, 5H), 2.21 - 2.15 (m, 2H), 2.03 (t, 2H), 1.91 - 1.796 (m, 2H), 1.51 - 1.42 (m, 4H), 1.21 - 1.12 (m, 2H), 0.88 (t, 3H).

[0671] LC-MS (ESI): m / z = 1523.8 [M+H] + .

[0672] Compound L-3-2:

[0673] 1H NMR (400 MHz, DMSO-d6) δ 8.71 (t, 1H), 8.59 (d, 1H), 8.26 (t, 1H), 8.14 (t, 1H), 8.06 (d, 1H), 7.97 (t, 1H), 7.77 (d, 2H), 7.32 (s, 1H), 7.29-7.14 (m, 5H), 6.99 (s, 2H), 6.50 (s, 1H), 5.61-5.52 (m, 1H), 5.46-5.38 (m, 2H), 5.23 (s, 2H), 4.80-4.74 (m, 1H), 4.73-4.66 (m, 2H), 4.60-4.54 (m, 1H), 4.53-4.44 (m, 1H), 4.05-3.99 (m, 1H), 3.79-3.65 (m, 5H), 3.62-3.54 (m, 3H), 3.52-3.44 (m, 28H), 3.41-3.33 (m, 5H), 3.23-3.12 (m, 3H), 3.06-2.98 (m, 1H), 2.81-2.72 (m, 1H), 2.70-2.54 (m, 6H), 2.42-2.33 (m, 5H), 2.17-2.07 (m, 2H), 2.03 (t, 2H), 1.93-1.78 (m, 1H), 1.52-1.40 (m, 4H), 1.22-1.11 (m, 2H), 0.88 (t, 3H).

[0674] LC-MS (ESI): m / z = 1523.8 [M+H] + .

[0675] Example L-4:

[0676] First Step: Take compound L-4A (500 mg, 1.88 mmol) as raw material, refer to the first step operation of Example L-3, to obtain compound L-4B (590 mg, 68.3%).

[0677] LC-MS (ESI): m / z = 459.3 [M+H] + .

[0678] Second Step: Take compound L-4B (500 mg, 1.09 mmol) as raw material, refer to the second step operation of Example L-3, to obtain compound L-4C (540 mg, 79.3%).

[0679] LC-MS (ESI): m / z = 625.2 [M+H] + .

[0680] Third Step: Take L-1 F (250 mg, 0.511 mmol) and compound L-4C (383 mg, 0.613 mmol) as materials, reference to the operation of the third step of example L-3, compound L-4D (355 mg, 74.7%) was obtained.

[0681] LC-MS (ESI): m / z = 928.4 [M-H] - .

[0682] Fourth Step: Take compound L-4D (300 mg, 0.323 mmol) as material, reference to the operation of the fourth step of example L-3, compound L-4-1 (124 mg, retention time 2.472 min, 28.5%) and compound L-4-2 (128 mg, retention time 2.522 min, 29.4%) were obtained.

[0683] Analytical HPLC condition: Instrument: CAS-05-LCMS-P; Column: C18. Mobile phase A, B composition: A is 0.0375% trifluoroacetic acid in water; B is 0.0185% trifluoroacetic acid in acetonitrile, gradient elution, B from 0% to 80% in 4 min, flow rate 1 mL / min.

[0684] Preparative HPLC separation condition: Instrument: CAS-05-Semi-prep S; Column: SunFire® Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A 0.1% trifluoroacetic acid in water; B acetonitrile, gradient elution, B from 0% to 60% in 18 min, flow rate 30 mL / min. Elution time 25 min.

[0685] Compound L-4-1:

[0686] 1H NMR (400 MHz, DMSO-d6) δ 8.67 (t, 1H), 8.54 (d, 1H), 8.28 (t, 1H), 8.15 (t, 1H), 8.06 (d, 1H), 7.98 (t, 1H), 7.79 (t, 1H), 7.73 (d, 1H), 7.30 (s, 1H), 7.28 - 7.13 (m, 5H), 6.99 (s, 2H), 6.50 (s, 1H), 5.63 - 5.54 (m, 1H), 5.46 - 5.38 (m, 2H), 5.15 - 5.05 (m, 2H), 4.79 - 4.66 (m, 3H), 4.58 - 4.43 (m, 2H), 3.98 (d, 1H), 3.79 - 3.65 (m, 5H), 3.64 - 3.55 (m, 3H), 3.52 - 3.45 (m, 12H), 3.41 - 3.36 (m, 5H), 3.26 - 3.10 (m, 3H), 3.06 - 2.98 (m, 1H), 2.81 - 2.67 (m, 3H), 2.67 - 2.59 (m, 3H), 2.42 - 2.34 (m, 5H), 2.24 - 2.14 (m, 2H), 2.06 - 1.99 (m, 2H), 1.93 - 1.81 (m, 2H), 1.52 - 1.41 (m, 4H), 1.23 - 1.11 (m, 2H), 0.88 (t, 3H).

[0687] LC-MS (ESI): m / z = 1347.7 [M+H] + .

[0688] Compound L-4-2:

[0689] 1H NMR (400 MHz, DMSO-d6) δ 8.73 (t, 1H), 8.59 (d, 1H), 8.27 (t, 1H), 8.16 (t, 1H), 8.07 (d, 1H), 7.99 (t, 1H), 7.79 (t, 1H), 7.73 (d, 1H), 7.31 (s, 1H), 7.26 (d, J = 7.6 Hz, 1H), 7.26 - 7.14 (m, 4H), 6.99 (s, 2H), 6.50 (s, 1H), 5.59 - 5.51 (m, 1H), 5.46 - 5.38 (m, 2H), 5.19 (s, 2H), 4.81 - 4.75 (m, 1H), 4.74 - 4.68 (m, 2H), 4.61 - 4.54 (m, 1H), 4.53 - 4.45 (m, 1H), 4.04 (d, 1H), 3.78 - 3.67 (m, 5H), 3.64 - 3.56 (m, 3H), 3.52 - 3.45 (m, 12H), 3.42 - 3.33 (m, 5H), 3.23 - 3.11 (m, 3H), 3.08 - 3.00 (m, 1H), 2.82 - 2.73 (m, 1H), 2.71 - 2.57 (m, 5H), 2.43 - 2.33 (m, 5H), 2.18 - 2.09 (m, 2H), 2.05 (t, 2H), 1.93 - 1.79 (m, 2H), 1.51 - 1.42 (m, 4H), 1.25 - 1.11 (m, 2H), 0.88 (t, 3H).

[0690] LC-MS (ESI): m / z = 1347.7 [M+H] + .

[0691] Example L-5:

[0692] First Step: Take L-1F (250 mg, 0.511 mmol) and compound L-5A (217 mg, 0.613 mmol) as raw materials, refer to the operation of the third step of Example L-3, to obtain compound L-5B (315 mg, 84.7%).

[0693] LC-MS (ESI): m / z = 727.3 [M-H] - .

[0694] Second Step: Take compound L-5B (200 mg, 0.274 mmol) as raw materials, refer to the operation of the fourth step of Example L-3, to obtain compound L-5-1 (68 mg, retention time 1.642 min, 21.6%) and compound L-5-2 (77 mg, retention time 1.696 min, 24.5%).

[0695] Analytical HPLC conditions: Instrument: CAS-05-LCMS-P; Column: C18. Mobile phase A, B composition: A 0.0375% trifluoroacetic acid in water; B 0.0185% trifluoroacetic acid in acetonitrile, gradient elution, B from 0% to 80% in 4 min, flow rate 1 mL / min.

[0696] Preparative HPLC separation conditions: Instrument: CAS-05-Semi-prep R; Column: SunFire® Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A 0.1% trifluoroacetic acid in water; B acetonitrile, gradient elution, B from 0% to 60% in 18 min, flow rate 25 mL / min. Elution time 25 min.

[0697] Compound L-5-1 :

[0698] LC-MS (ESI): m / z = 1146.6 [M+H] + .

[0699] 1 H NMR (400 MHz, DMSO-d6) δ 8.65 (t, 1H), 8.53 (d, 1H), 8.26 (t, 1H), 8.10 (t, 1H), 8.05 (d, 1H), 7.96 (t, 1H), 7.76 (d, 1H), 7.30 (s, 1H), 7.27 - 7.11 (m, 5H), 7.00 (s, 2H), 6.48 (s, 1H), 5.63 - 5.55 (m, 1H), 5.46 - 5.37 (m, 2H), 5.13 (s, 2H), 4.77 - 4.66 (m, 3H), 4.58 - 4.42 (m, 2H), 3.97 (d, 1H), 3.77 - 3.62 (m, 5H), 3.63 - 3.34 (m, 11H), 3.27 - 3.18 (m, 1H), 3.17 - 3.08 (m, 1H), 3.05 - 2.96 (m, 1H), 2.82 - 2.55 (m, 6H), 2.40 - 2.28 (m, 5H), 2.23 - 2.13 (m, 2H), 1.91 - 1.80 (m, 2H), 0.88 (t, 3H).

[0700] Compound L-5-2 :

[0701] 1H NMR (400 MHz, DMSO-d6) δ 8.72 (t, 1H), 8.59 (d, 1H), 8.25 (t, 1H), 8.12 (t, 1H), 8.07 (d, 1H), 7.97 (t, 1H), 7.77 (d, 1H), 7.32 (s, 1H), 7.27-7.14 (m, 5H), 7.00 (s, 2H), 6.50 (s, 1H), 5.60-5.51 (m, 1H), 5.46-5.37 (m, 2H), 5.23 (d, 2H), 4.82-4.63 (m, 3H), 4.61-4.40 (m, 2H), 4.05-3.96 (m, 1H), 3.79-3.63 (m, 5H), 3.63-3.40 (m, 11H), 3.21-3.10 (m, 2H), 3.07-2.97 (m, 1H), 2.81-2.76 (m, 1H), 2.69-2.57 (m, 5H), 2.41-2.28 (m, 5H), 2.13 (s, 2H), 1.91-1.79 (m, 2H), 0.87 (t, 3H).

[0702] LC-MS (ESI): m / z = 1146.6 [M+H] + .

[0703] Example L-6:

[0704] First Step: Take compound L-6A (1.00 g, 5.64 mmol) as raw material, refer to the first step of Example L-3, compound L-6B (1.96 g, 93.8%) is obtained.

[0705] LC-MS (ESI): m / z = 371.2 [M+H] + .

[0706] Second Step: Take compound L-6B (500 mg, 1.35 mmol) as raw material, refer to the second step of Example L-3, compound L-6C (570 mg, 78.7%) is obtained.

[0707] LC-MS (ESI): m / z = 537.2 [M+H] + .

[0708] Third Step: Take L-1F (200 mg, 0.409 mmol) and compound L-6C (263 mg, 0.491 mmol) as raw materials, refer to the third step of Example L-3, compound L-6D (230 mg, 66.9%) is obtained.

[0709] LC-MS (ESI): m / z = 840.4 [M-H]- .

[0710] Fourth step: using compound L-6D (130 mg, 0.154 mmol) as raw material, reference to the operation of the fourth step of example L-3, compound L-6-1 (46 mg, retention time 2.453 min, 23.7%) and compound L-6-2 (52 mg, retention time 2.508 min, 26.7%) were obtained.

[0711] Analytical HPLC conditions: instrument: CAS-05-LCMS-P; column: C18. Mobile phase A, B composition: A 0.0375% trifluoroacetic acid aqueous solution; B 0.0185% trifluoroacetic acid acetonitrile, gradient elution, B from 0% to 80% in 4 min, flow rate 1 mL / min.

[0712] Preparative HPLC separation conditions: instrument: CAS-05-Semi-prep K; column: SunFire@Prep C18 (19 mm x 250 mm). Mobile phase A, B composition: A water; B acetonitrile, gradient elution, B from 0% to 60% in 18 min, flow rate 30 mL / min. Elution time 25 min.

[0713] Compound L-6-1:

[0714] 1 H NMR (400 MHz, DMSO-d6) δ 8.66 (t, 1H), 8.54 (d, 1H), 8.27 (t, 1H), 8.13 (t, 1H), 8.06 (d, 1H), 7.98 (t, 1H), 7.76 (d, 2H), 7.31 (s, 1H), 7.28 - 7.13 (m, 5H), 6.99 (s, 2H), 6.49 (s, 1H), 5.63 - 5.56 (m, 1H), 5.42 (s, 2H), 5.13 (s, 2H), 4.73 - 4.67 (m, 3H), 4.58 - 4.43 (m, 2H), 4.01 - 3.94 (m, 1H), 3.78 - 3.66 (m, 5H), 3.64 - 3.55 (m, 3H), 3.51 - 3.42 (m, 4H), 3.41 - 3.34 (m, 4H), 3.25 - 3.10 (m, 4H), 3.04 - 2.97 (m, 1H), 2.81 - 2.60 (m, 6H), 2.42 - 2.34 (m, 5H), 2.24 - 2.15 (m, 2H), 2.03 (t, 2H), 1.92 - 1.81 (m, 2H), 1.51 - 1.41 (m, 4H), 1.26 - 1.11 (m, 2H), 0.88 (t, 3H).

[0715] LC-MS (ESI): m / z = 1259.6 [M+H] + .

[0716] Compound L-6-2:

[0717] 1 H NMR (400 MHz, DMSO-d6) d 8.71 (t, 1H), 8.58 (d, 1H), 8.25 (t, 1H), 8.14 (t, 1H), 8.06 (d, 1H), 7.97 (t, 1H), 7.80-7.73 (m, 2H), 7.32 (s, 1H), 7.29-7.13 (m, 5H), 6.98 (s, 2H), 6.49 (s, 1H), 5.58-5.50 (m, 1H), 5.42 (s, 2H), 5.22 (s, 2H), 4.81-4.74 (m, 1H), 4.73-4.67 (m, 2H), 4.60-4.44 (m, 2H), 4.05-3.99 (m, 1H), 3.79-3.66 (m, 5H), 3.64-3.54 (m, 3H), 3.50-3.42 (m, 4H), 3.40-3.32 (m, 4H), 3.23-3.09 (m, 4H), 3.07-2.97 (m, 1H), 2.81-2.71 (m, 1H), 2.69-2.56 (m, 5H), 2.41-2.34 (m, 5H), 2.17-2.09 (m, 2H), 2.10-1.98 (m, 2H), 1.91-1.81 (m, 2H), 1.50-1.41 (m, 4H), 1.26-1.10 (m, 2H), 0.88 (t, 3H).

[0718] LC-MS (ESI): m / z = 1259.6 [M+H] + .

[0719] Example L-7:

[0720] First Step: Take L-1F (250 mg, 0.511 mmol) and compound L-7A (190 mg, 0.613 mmol) as raw materials, refer to the operation of the third step of Example L-3, to obtain compound L-7B (190 mg, 54.3%).

[0721] LC-MS (ESI): m / z = 683.3 [M-H] - .

[0722] Second step: Take compound L-7B (190 mg, 0.277 mmol) as raw material, reference to the operation of the fourth step of example L-3, to obtain compound L-7-1 (104 mg, retention time 2.414 min, 34.0%) and compound L-7-2 (112 mg, retention time 2.472 min, 36.6%).

[0723] Analytical HPLC conditions: Instrument: CAS-05-LCMS-P; Column: C18. Mobile phase A, B composition: A is 0.0375% trifluoroacetic acid aqueous solution; B is 0.0185% trifluoroacetic acid acetonitrile solution, gradient elution, B from 0% to 80% in 4 min, flow rate 1 mL / min.

[0724] Preparative HPLC separation conditions: Instrument: CAS-05-Semi-prep K; Column: SunFire@Prep C18(19mm x 250mm). Mobile phase A, B composition: A water; B acetonitrile, gradient elution, B from 0% to 60% in 18 min, flow rate 25 mL / min. Elution time 25 min.

[0725] Compound L-7-1:

[0726] 1 H NMR (400 MHz, DMSO-d6) δ 8.66 (t, 1H), 8.54 (d, 1H), 8.27 (t, 1H), 8.11-8.02 (m, 2H), 7.97 (t, 1H), 7.75 (d, 1H), 7.30 (s, 1H), 7.26-7.12 (m, 5H), 6.99 (s, 2H), 6.49 (s, 1H), 5.63-5.55 (m, 1H), 5.46-5.35 (m, 2H), 5.11 (s, 2H), 4.77-4.66 (m, 3H), 4.57-4.43 (m, 2H), 4.00-3.94 (m, 1H), 3.86-3.60 (m, 6H), 3.58-3.49 (m, 4H), 3.48-3.43 (m, 2H), 3.27-3.08 (m, 2H), 3.04-2.97 (m, 1H), 2.80-2.57 (m, 6H), 2.39m 2.27 (m, 5H), 2.22m 2.14 (m, 2H), 1.90m 1.80 (m, 2H), 0.88 (t, 3H).

[0727] LC-MS (ESI): m / z = 1102.4 [M+H] + .

[0728] Compound L-7-2:

[0729] 1 H NMR (400 MHz, DMSO-d6) δ 8.72 (t, 1H), 8.59 (d, 1H), 8.26 (t, 1H), 8.15-8.03 (m, 2H), 7.97 (t, 1H), 7.75 (d, 1H), 7.31 (s, 1H), 7.28-7.13 (m, 5H), 6.99 (s, 2H), 6.50 (s, 1H), 5.59-5.51 (m, 1H), 5.42 (s, 2H), 5.21 (s, 2H), 4.81-4.66 (m, 3H), 4.61-4.43 (m, 2H), 4.03 (d, 1H), 3.75-3.64 (m, 5H), 3.59-3.50 (m, 5H), 3.49-3.42 (m, 2H), 3.24-3.08 (m, 2H), 3.06-2.99 (m, 1H), 2.82-2.72 (m, 1H), 2.69-2.56 (m, 5H), 2.41-2.35 (m, 3H), 2.32 (t, 2H), 2.19-2.09 (m, 2H), 1.90-1.79 (m, 2H), 0.88 (t, 3H).

[0730] LC-MS (ESI): m / z = 1102.4 [M+H] + .

[0731] 3. Preparation of antibody drug conjugates

[0732] Example: Coupling preparation of Raludotatug-L-1-1

[0733] Take 10 mg of Raludotatug antibody (purchased from MCE), add 20 eq. of TCEP, dilute to 5 mg / ml with PBS, reduce at 37°C for 2 hours. After cooling to room temperature, add 13 times the molar ratio of compound L-1-1, DMSO ratio is 10% of the total volume, react at 25°C for 2 hours. Purify by 30KD ultrafiltration concentration tube, 0.22μm sterilization filter to get 10mg product, MS-DAR is 8.00, SEC purity is >99%, storage buffer is 25mM His-HOAc, pH 5.5.

[0734] Example: Coupling preparation of Raludotatug-L-2

[0735] Take 10 mg of Raludotatug antibody, add 20 eq. of TCEP, dilute to 5 mg / ml with PBS, reduce at 37°C for 2 hours. After cooling to room temperature, add 13 times the molar ratio of compound L-2, DMSO ratio is 10% of the total volume, react at 25°C for 2 hours. Purify by 30KD ultrafiltration concentrator, 0.22μm sterile filtration to get 12 mg of product, MS-DAR is 8.00, SEC purity is 99.17%, storage buffer is 25mM His-HOAc, pH 5.5.

[0736] Example: Coupling preparation of Trastuzumab-L-1-1

[0737] Take 10 mg of Trastuzumab antibody, add 20 eq. of TCEP, dilute to 5 mg / ml with PBS, reduce at 37°C for 2 hours. After cooling to room temperature, add 13 times the molar ratio of compound L-1-1, DMSO ratio is 10% of the total volume, react at 25°C for 2 hours. Purify by 30KD ultrafiltration concentrator, 0.22μm sterile filtration to get 10 mg of product, storage buffer is 25mM His-HOAc, pH 5.5.

[0738] Example: Coupling preparation of Sacituzumab-L-1-1

[0739] Take 10 mg of Sacituzumab antibody, add 20 eq. of TCEP, dilute to 5 mg / ml with PBS, reduce at 37°C for 2 hours. After cooling to room temperature, add 13 times the molar ratio of compound L-1-1, DMSO ratio is 10% of the total volume, react at 25°C for 2 hours. Purify by 30KD ultrafiltration concentrator, 0.22μm sterile filtration to get 11 mg of product, storage buffer is 25mM His-HOAc, pH 5.5.

[0740] Example: Coupling preparation of Labetuzumab-L-1-1

[0741] Labetuzumab antibody was left to stand at room temperature. After it was brought to room temperature, the volume of antibody required was calculated. At the same time, a 10 mM TCEP aqueous solution was prepared, and the 10 mM TCEP aqueous solution was mixed with the antibody and incubated in a 37 °C metal bath for 2 hours. Compound L-1-1 was dissolved in DMSO. After the antibody was brought to room temperature, the DMSO solution was added in advance, followed by standing for 5 minutes, and the reaction solution was controlled at room temperature. The volume of 10 mM compound L-1-1 solution required to be added was calculated according to the LP / antibody molar ratio of 13:1, and the DMSO ratio was maintained at 10% (v / v). After thorough mixing, it was incubated in a 25 °C metal bath for 2 hours. Subsequently, a 10% (w / v) activated carbon powder solution was prepared, and PBS solution was washed 3 times to obtain an antibody conjugate solution. In order to purify the antibody conjugate drug, 10% (v / v) activated carbon solution was added to the antibody and small molecule conjugate solution, and thoroughly mixed at room temperature for 1 hour. After the mixing was completed, the activated carbon powder was centrifuged at low speed, and filtered with a 0.22 μm PES (Merck) filter, and the filtered solution was transferred to a 30KDa Amicon (Merck), and centrifuged at 2000g for ultrafiltration exchange 3 to 5 times, and finally the antibody conjugate solution was stored in 25 mM His-HOAc, pH 5.5. Finally, the antibody conjugate drug DAR = 8, SEC purity 99.51% was obtained.

[0742] Example: Coupling preparation of Raludotatug-L-5-1

[0743] 10 mg of Raludotatug antibody (purchased from MCE) was added with 20 eq. of TCEP, diluted with PBS to 5 mg / ml, and reduced at 37 °C for 2 hours. After cooling to room temperature, 13 times the molar ratio of compound L-5-1 was added, and the DMSO ratio was 10% of the total volume, and the reaction was carried out at 25 °C for 2 hours. After purification by 30KD ultrafiltration concentration tube and 0.22 μm sterilization filtration, 6 mg of product was obtained, the MS-DAR was 7.92, the SEC purity was 98.28%, and the storage buffer was 25 mM His-HOAc, pH 5.5.

[0744] Biological test evaluation

[0745] The following test examples further illustrate the application, but are not meant to limit the scope of the application.

[0746] 1. Cell proliferation inhibition activity

[0747] OVCAR3 (ATCC, HTB-161), A2780 (Cobioer, CBP60283), OVCAR8, OVKATE, HCC1806, SK-CO-1, MKN45, HCT116, MOLT4, EOL-1, HT29 cells were cultured in a 37℃, 5% CO2 incubator for 48-72h. Cells were trypsinized and counted, and the cell concentration was adjusted to the appropriate cell density, 100μL of cells were inoculated into the bottom transparent 96-well plate (Greiner, 655090) per well, and transferred to a 37℃, 5% CO2 incubator for overnight culture. The next day, 50μL of gradient-diluted culture medium containing different concentrations of test compounds was added to each well, and 2 replicate wells were set for each concentration. At the same time, DMSO solvent control and negative control groups were set, and the culture was continued in a 37℃, 5% CO2 incubator for 5-7 days. After incubation, the culture medium was removed from each well Detection reagent (Promega, G7558) was restored to room temperature, and 50μL was added to each well Detection reagent, and the 96-well plate was placed on a shaker for 10min (the whole process should be operated in the dark), and the fluorescence signal value LUM of each well was detected using the Luminescence module of the PHERAstar FSX multifunctional enzyme labeler (BMG LRBTECH). The inhibition rate of the test compound was calculated by the formula: The IC 50 Values of the test compounds were calculated using the Graphpad software log(inhibitor) vs.response--Variable slope(four parameters) equation for fitting analysis. The IC 10 Values of the test compounds were calculated using the Graphpad software log(inhibitor) vs.response--Variable slope(four parameters) equation for fitting analysis. The IC 50 Values of the test compounds were calculated using the Graphpad software log(inhibitor) vs.response--Variable slope(four parameters) equation for fitting analysis. The IC

[0748] Table 1.1 Inhibition of the proliferation of OVCAR3 cells by compounds

[0749] Note: A < 5nM, 5nM ≤ B < 10nM, 10nM ≤ C < 50nM

[0750] Table 1.2 Inhibition of the proliferation of A2780 cells by compounds

[0751] Note: A < 5nM, 5nM ≤ B < 10nM, 10nM ≤ C < 50nM

[0752] Table 1.3 Inhibition of the proliferation of OVCAR8 cells by compounds

[0753] Table 1.4 Inhibition of proliferation of OVKATE cells by compounds

[0754] Table 1.5 Inhibition of proliferation of HCC1806 cells by compounds

[0755] Table 1.6 Inhibition of proliferation of SK-CO-1 cells by compounds

[0756] Table 1.7 Inhibition of proliferation of MKN45 cells by compounds

[0757] Table 1.8 Inhibition of proliferation of HCT116 cells by compounds

[0758] Note: The structure of the control Labetuzumab-GGFG-Dxd is:

[0759] The structure of Dxd is:

[0760] Conclusion: The compounds of the present application, such as the compounds of the examples, have significant inhibitory effects on the proliferation of OVCAR3, A2780, OVCAR8, OVKATE, HCC1806, SK-CO-1, MKN45, HCT116 cells. The IC 50 of compound 1-1 and compound 1-2 on OVCAR3 is 0.101 nM and 0.086 nM, and the IC 50 of compound 3-1 on OVCAR3 is 0.093 nM.

[0761] 2. TOP1 enzymatic inhibition test

[0762] Compounds were dispensed into 96-well plates using an Echo. (The highest concentration was 30 micromolar per liter, with a 3-fold dilution, for a total of 9 dose gradients), 15 microliters of reaction buffer (25 millimolar per liter Tris-HCl, pH 7.4, 58 millimolar per liter KCl, 0.25 millimolar per liter DTT, 5 millimolar per liter MgCl2, 0.25 millimolar per liter EDTA, 15 micrograms per milliliter bovine serum albumin) containing DNA topoisomerase I (2 units) was added to each well. Compounds and DNA topoisomerase I were incubated at 37 degrees Celsius for 30 minutes. 5 microliters of supercoiled DNA (0.25 micrograms) was added to each of the 96 wells, and the 96-well plate was incubated at 37 degrees Celsius for 1 hour. 125 microliters of 1X concentration of H19 dye was added to each of the 96 wells. This was then incubated at room temperature for 5 minutes. Fluorescence intensity was measured at 520 nanometers using an excitation wavelength of 485 nanometers.

[0763] Inhibition calculation formula: %inhibition = 100*(Ssample-Slow) / (SHC-SLC)

[0764] Fitting formula: R_IC50: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope))

[0765] Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope + log((Top-Bottom) / (Fifty-Bottom)-1))

[0766] Conclusion: The compounds of the present application, such as the example compounds, have significant inhibitory activity on TOP1 enzyme.

[0767] 3. Mouse pharmacokinetic test

[0768] 3.1 Test animals: Male Balb / c mice, 20-25 g, 3 per compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0769] 3.2 Test design: On the test day, the Balb / c mice were randomly divided into groups according to body weight. Fasting for 12-14 hours without water restriction 1 day before administration, and feeding 4 hours after administration.

[0770] Table 3.1 Dosing information

[0771] Note: Intravenous administration vehicle: 5% DMA + 5% HS15 + 90% Saline

[0772] The blood was taken from the orbit before and after administration, 0.06 mL was placed in an EDTA K2 centrifuge tube, centrifuged at 5000 rpm at 4 DEG C for 10 min, and the plasma was collected. The blood sampling time points of the intravenous group were 0, 5, 15, 30 min, 1, 2, 4, 6, 8, and 24 h. Before analysis and detection, all samples were stored at -80 DEG C, and the samples were quantitatively analyzed by LC-MS / MS.

[0773] Table 3.2 Pharmacokinetic parameters of the test compounds in mouse plasma

[0774] Conclusion: The compound of the present application, for example, the compound of the examples, has good pharmacokinetic characteristics in mice.

[0775] 4. Rat pharmacokinetic test

[0776] 4.1 Test animals: male SD rats, about 220 g, 6-8 weeks old, 3 rats per compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0777] 4.2 Test design: On the test day, the SD rats were randomly divided according to the body weight. The rats were fasted for 12-14 h without water 1 day before administration, and were fed 4 h after administration.

[0778] Table 4.1 Dosing information

[0779] Note: Intravenous administration vehicle: 5% DMA + 5% HS15 + 90% saline

[0780] The blood was taken from the orbit before and after administration, 0.06 mL was placed in an EDTA K2 centrifuge tube, centrifuged at 5000 rpm at 4 DEG C for 10 min, and the plasma was collected. The blood sampling time points of the intravenous group were 0, 5, 15, 30 min, 1, 2, 4, 6, 8, and 24 h. Before analysis and detection, all samples were stored at -80 DEG C, and the samples were quantitatively analyzed by LC-MS / MS.

[0781] Table 4.2 Pharmacokinetic parameters of the test compounds in rat plasma

[0782] Conclusion: The compound of the present application, for example, the compound of the examples, has good pharmacokinetic characteristics in rats.

[0783] 5. Beagle dog pharmacokinetic test

[0784] 5.1 Test animals: male beagle dogs, about 8-11 kg, 3 dogs per compound, purchased from Beijing Mas Biotechnology Co., Ltd.

[0785] 5.2 Test method: On the test day, the beagle dogs were randomly grouped according to the body weight. 12-14 h before the administration, the dogs were fasted but not water-deprived, and 4 h after the administration, the dogs were fed.

[0786] Before and after the administration, 1 ml of blood was taken through the jugular vein or the limb vein and placed in an EDTA K2 centrifuge tube. Centrifugation was performed at 5000 rpm and 4℃ for 10 min, and the plasma was collected. The blood sampling time points of the intravenous group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, and 48 h. Before analysis and detection, all the samples were stored at-80℃, and the samples were subjected to quantitative analysis by using LC-MS / MS.

[0787] Conclusion: The compound of the present application, for example, the compound of the examples, has good in-dog pharmacokinetic characteristics.

[0788] 6. Pharmacokinetic test in monkeys

[0789] 6.1 Test animals: Male cynomolgus monkeys, 3-5 kg, 3-6 years old, 2 monkeys per compound. Purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0790] 6.2 Test method: On the test day, the monkeys were randomly grouped according to the body weight. 14-18 h before the administration, the monkeys were fasted but not water-deprived, and 4 h after the administration, the monkeys were fed.

[0791] Table 5.1 Dosing information

[0792] Note: Intravenous administration solvent: 10% DMA + 5% HS15 + 85% saline

[0793] Before and after the administration, 1.0 mL of blood was taken through the limb vein and placed in an EDTA K2 centrifuge tube. Centrifugation was performed at 5000 rpm and 4℃ for 10 min, and the plasma was collected. The blood sampling time points of the intravenous group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, and 48 h. Before analysis and detection, all the samples were stored at-80℃, and the samples were subjected to quantitative analysis by using LC-MS / MS.

[0794] Table 5.2 Pharmacokinetic parameters of the test compound in the plasma of monkeys

[0795] Conclusion: The compound of the present application, for example, the compound of the examples, has good in-monkey pharmacokinetic characteristics.

[0796] 7. Plasma stability test

[0797] In this experiment, the plasma of five species, i.e., human, monkey, dog, rat, and mouse, was used to evaluate the plasma stability of the compound.

[0798] Prepare plasma samples with concentration level of 1000 ng / mL, aliquot into EP tubes with time points of 0 h and 6 h; wherein 0 h samples are directly added into acetonitrile solution containing internal standard, and 6 h samples are added into acetonitrile solution containing internal standard after being placed at 37℃ for corresponding time. The concentration of the test substance in the sample is detected by LC-MS / MS method, and the residual rate is calculated by the ratio of the peak area of the test substance and internal standard in the time point sample to that in the zero time sample.

[0799] Conclusion: The compound of the present application, for example, the compound of the examples, has lower plasma stability, can effectively reduce the plasma toxin exposure level, and reduce the toxic side effects related to the compound.

[0800] 8. Caco2 permeability test

[0801] The test uses single layer Caco-2 cells, and three parallel incubations are used in 96-well Transwell plates. Transport buffer solution (HBSS, 10 mM HEPES, pH 7.4±0.05) containing the compound of the present application (2 μM) or control compounds digoxin (10 μM), nadolol (2 μM) and metoprolol (2 μM) is added to the dosing end hole of the top side or the base side. The receiving end hole is added with DMSO-containing transport buffer solution. After incubation at 37±1℃ for 2 hours, the cell plate is taken out and an appropriate amount of sample is taken from the top and bottom of each new 96-well plate. Then acetonitrile containing internal standard is added to precipitate the protein. The sample is analyzed by LC MS / MS and the concentration of the compound of the present application and the control compound is determined. The concentration data is used to calculate the apparent permeability coefficient of the transport from the top side to the base side of the single layer cells, and the base side to the top, and to calculate the efflux rate. The integrity of the single layer cells after 2 hours of incubation is evaluated by the leakage of fluorescein.

[0802] Table 6 Caco2 permeability

[0803] Conclusion: The compound of the present application, for example, the compound of the examples, has good permeability and low efflux rate.

[0804] 9. hERG potassium ion channel effect test

[0805] 9.1. Experimental platform: electrophysiological hand-held patch clamp system

[0806] 9.2. Cell line: Chinese hamster ovary (CHO) cell line stably expressing hERG potassium ion channel

[0807] 9.3, Experimental method: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channel were used to record hERG potassium current by whole-cell patch clamp technique at room temperature. Glass microelectrode was pulled from glass electrode blank (BF150-86-10, Sutter) by a puller, and the tip resistance was about 2-5 MΩ after filling the electrode with internal solution. The glass microelectrode was inserted into the amplifier probe and then connected to the patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by computer through pClamp 10 software, and the sampling frequency was 10 kHz and the filter frequency was 2 kHz. After obtaining the whole-cell recording, the cell was clamped at -80 mV, and the step voltage for inducing hERG potassium current (IhERG) was given from -80 mV to +20 mV for 2 s, and then repolarized to -50 mV for 1 s, and then returned to -80 mV. This voltage stimulation was given every 10 s, and after the hERG potassium current was determined to be stable (at least 1 min), the drug administration process was started. Each test concentration of the compound was given for at least 1 min, and at least 2 cells were tested for each concentration (n≥2).

[0808] 9.4, Data processing: pClamp 10, GraphPad Prism 5 and Excel software were used for data analysis and processing. The inhibition degree of different compound concentrations on hERG potassium current (peak value of hERG tail current induced at -50 mV) was calculated by the following formula:

[0809] Inhibition% = [1-(I / Io)]x100%

[0810] Wherein, Inhibition% represents the inhibition percentage of the compound on hERG potassium current, and I and Io represent the amplitude of hERG potassium current after and before drug administration, respectively.

[0811] The IC50 of the compound was calculated by fitting the following equation using GraphPad Prism 5 software:

[0812] Y = Bottom + (Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0813] Wherein, X is the Log value of the test concentration of the test product, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0814] Conclusion: The compound of the present application has no obvious inhibitory effect on hERG potassium channel current.

[0815] 10, CYP450 enzyme inhibition test

[0816] The purpose of this study is to evaluate the effect of the test substance on the activity of five isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) of human liver microsomal cytochrome P450 (CYP) by using in vitro test system. The specific probe substrates of CYP450 isozymes are incubated with human liver microsomes and different concentrations of test substance respectively, and reduced nicotinamide adenine dinucleotide phosphate (NADPH) is added to start the reaction. After the reaction is completed, the specific substrate metabolites are quantitatively detected by treating the sample and using liquid chromatography-tandem mass spectrometry (LC-MS / MS) method, the change of CYP enzyme activity is determined, the IC50 value is calculated, and the inhibition potential of the test substance on each CYP enzyme subtype is evaluated.

[0817] Conclusion: The compound of the present application has no obvious inhibitory activity on the five isozymes of human liver microsomal cytochrome P450 (CYP).

[0818] 11. Liver microsomal stability test

[0819] In this experiment, human, monkey, dog, rat and mouse liver microsomes of five species were used as in vitro models to evaluate the metabolic stability of the test substance.

[0820] At 37℃, 1 μM of the test substance is incubated with microsomal protein and coenzyme NADPH, and the reaction is terminated by adding ice-cold acetonitrile containing internal standard at a certain time (5, 10, 20, 30, 60 min). The concentration of the test substance in the sample is detected by LC-MS / MS method to obtain the residual rate of the drug after 60 min of incubation.

[0821] Table 7 Liver microsomal metabolic stability

[0822] Conclusion: The compound of the present application has good metabolic stability in human liver microsomes.

[0823] 12. ADC bystander killing test

[0824] 293T-CDH6 (Kyinno Bio, KC-3200), 293T (ATCC) cells were cultured in EMEM complete medium at 37°C, 5% CO2 for 48-72h. Cells were trypsinized and counted, and the cell concentration was adjusted to the appropriate cell density, 4200 cells / well, 600 μL / well for 293T-CDH6 cells, and 300 cells / well, 100 μL / well for 293T cells were seeded into the lower chamber of a 24-well transwell plate (Corning, 3421), and the 24-well transwell plate (Corning, 3421) was transferred to a 37°C, 5% CO2 incubator for overnight culture. The next day, 150 μL of the medium containing different concentrations of the test compound was added to the upper chamber, and 100 μL of the medium containing different concentrations of the test compound was added to the lower chamber. At the same time, a DMSO solvent control group was set up, and the culture was continued in a 37°C, 5% CO2 incubator for 4 days.

[0825] After the end of the culture, the 24-well transwell plate was taken out The CellTiter-Glo® reagent (Promega, G7558) was restored to room temperature. A clean 24-well plate (NEST, 702001) was prepared, and the upper chamber was placed in the clean 24-well plate, 150 μL of the CellTiter-Glo® reagent was added to each well. The medium in the lower chamber was removed by aspiration, 100 μL of the CellTiter-Glo® reagent was added to each well, and the 24-well plate was placed on a shaker for 10 min (the whole process needs to be operated in the dark). After shaking, 100 μL of cell lysate was taken from each well and placed in a clean black-bottom transparent 96-well plate (GREINER, 655090). The fluorescence signal value LUM of each well was detected using the Luminescence module of the PHERAstar FSX multifunctional enzyme labeler (BMG LABTECH). The inhibition rate of the test compound in the co-culture and only negative cells was calculated by the formula inhibition rate % = (1-(LUM compound / (LUM DMSO solvent control group))) x 100%. The Graphpad software was used for fitting analysis, and the inhibition rate of the test compound on 293T-CDH6 and 293T cells was calculated. The vertical coordinate of the data was the inhibition rate percentage, and the horizontal coordinate was the sample concentration. The results are shown in Figure 1.

[0826] Conclusion: Raludotatug-L-1-1 has stronger bystander killing activity than Raludotatug-GGFG-Dxd.

[0827] 13. Pharmacodynamic evaluation scheme of OVCAR3 subcutaneous tumor model

[0828]

[0829] ​Experimental reagents: Human ovarian cancer OVCAR3 cells were purchased from ATCC, RPMI-1640 medium was purchased from Gibco (item number 22400-089), fetal bovine serum was purchased from HyClone (item number SH30406.05), penicillin-streptomycin was purchased from Gibco (item number 15140-122), bovine insulin was purchased from MCE (item number HY-P1156), 0.5% trypsin-EDTA was purchased from Gibco (item number 15400-054), PBS was purchased from Sangon Biotech (item number E607008-0500), Matrigel (matrix glue) was purchased from Corning (item number 354234).

[0830] Experimental methods: Animal information: Balb / c nude mice, female, 5-7 weeks, body weight about 15-18 grams, Sichuan Weitong Lihua Experimental Animal Technology Co., Ltd., the mice were raised in SPF level environment, each cage was sent separately with exhaust, all animals could freely obtain standard certified commercial laboratory diet and free drinking water.

[0831] Cell culture: Human ovarian cancer OVCAR3 cell line was cultured in vitro, the culture conditions were 20% fetal bovine serum, 1% penicillin-streptomycin, 10 μg / mL bovine insulin added in RPMI-1640, 37 ℃, 5% CO2 incubator. When the cells adhered, the confluence reached 80%-90%, the passage ratio was 1:2-1:3, and the cells were passaged once every 3 days or 4 days, and placed in a 37 ℃, 5% CO2 cell culture box for culture, and the cell expansion cycle was about 20 days. When the number of cells reached the requirement, the cells were collected and counted.

[0832] Cell inoculation: 0.2 mL OVCAR3 cell suspension (containing 3x10 6 cells, PBS volume ratio 1:1) was subcutaneously inoculated in the armpit of each mouse.

[0833] Group administration: When the tumor grew to 100-150 mm 3 , the mice were randomly grouped for administration according to the tumor volume, and the grouping day was Day 0.

[0834] Tumor measurement and experimental indicators: The tumor diameter was measured twice a week with a vernier caliper. The formula for calculating the tumor volume was: V=0.5a x b, a and b represented the long diameter and short diameter of the tumor respectively. The body weight of the mice was measured twice a week. The antitumor effect of the test drug was evaluated by tumor growth inhibition rate TGI (%).

[0835] Tumor growth inhibition rate calculation formula:

[0836] TGI (%) = [1-(T Vt -T V0 ) / (CVt - C V0 )] x 100%

[0837] T Vt is the average tumor volume at the end of the dosing group; T V0 is the average tumor volume at the grouping of the dosing group; C Vt is the average tumor volume at the end of the Vehicle group; C V0 is the average tumor volume at the grouping of the Vehicle group. T Vt and C Vt Take the same day data.

[0838] Statistical analysis: Data are expressed as mean values. Statistical methods were analyzed by Two-Way ANOVA and Dunnett's Multiple Comparisons Test. If P value < 0.05, it is considered that there is a significant difference between groups. The results of the experiment are shown in Figure 2 and Table 8 below.

[0839] Table 8 Tumor volume in OVCAR3 subcutaneous tumor model

[0840] Conclusion: In the mouse subcutaneous transplanted tumor OVCAR3 model, ADC Raludotatug-L-1-1, Raludotatug-L-2 and Raludotatug-L-5-1 all have significant inhibitory effect on tumor growth (P < 0.0001) at a dose of 3 mg / kg after single intravenous administration.

Claims

1. A compound of Formula (IV-1), Formula (IV-2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: wherein E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, Preferred -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-、-O-CH2-C(O)NR d -CHR b -C(O)-、 R b C replaced by 1-4 Rs 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl, preferably R b Cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups; R d is hydrogen, deuterium, C 1-6 alkyl, preferably R d is hydrogen, deuterium, C 1-3 alkyl; or R b and R d with the carbon atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxy, oxo, preferably R b and R d with the carbon atom to which it is attached forms an azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxy, oxo; R a is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C 1-6 alkylaminoalkyl, C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-10 aryl or 5- to 6-membered heteroaryl, wherein said C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-10 aryl or 5- to 6-membered heteroaryl are optionally further substituted with 1 to 4 R; R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene; n1 is 1 or 2; n2 is 1 or 2; m1 is 1, 2, 3; m2 is 1, 2, 3.

2. The compound, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, which satisfies one or more of the following conditions: (1) R a is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-8 aryl or 5-6 membered heteroaryl is optionally further substituted with 1-4 R; (2) R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl, wherein the C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, phenyl, or 5-6 membered heteroaryl is optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene, preferably deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, (3) m1 and m2 are each independently 1, 2.

3. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of claim 1, wherein The E is 4. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of any one of claims 1-3, wherein, one of the structures selected from Table I.

5. A linker-drug conjugate of Formula (L-IV-1), Formula (L-IV-2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: wherein, E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, Preferred -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-、-O-CH2-C(O)NR d -CHR b -C(O)-、 R b C replaced by 1-4 Rs 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl, preferably R b Cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups; R d is hydrogen, deuterium, C 1-6 alkyl, preferably R d is hydrogen, deuterium, C 1-3 alkyl; or R b and R d with the carbon atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxy, oxo, preferably R b and R d with the carbon atom to which it is attached forms an azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxy, oxo; R a is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C 1-6 alkylaminoalkyl, C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-10 aryl or 5- to 6-membered heteroaryl, wherein said C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-10 aryl or 5- to 6-membered heteroaryl are optionally further substituted with 1 to 4 R, preferably hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-8 aryl or 5- to 6-membered heteroaryl, wherein said C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-8 aryl or 5- to 6-membered heteroaryl are optionally further substituted with 1 to 4 R; R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene, preferably deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, n1 is 1 or 2; n2 is 1 or 2; m1 is 1, 2, 3; m2 is 1, 2, 3; L is L a -L2-L3-L4-, wherein: L a is (maleimide-N-yl)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl), or -C 1-6 heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl containing 1-3 atoms selected from N, O, or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy; L2is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, where p is an integer from 0 to 20; L3 is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy; L4is a bond, -NR L4 (CR L5 R L6 ) q - C(O)NR L4 (CH2) q - wherein q is an integer from 0 to 6; R L2 and R L4 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy; R L5 and R L6 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl.

6. The linker-drug conjugate of claim 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The L is the following structure: where p is an integer from 1 to 10.

7. The linker-drug conjugate, stereoisomer, or pharmaceutically acceptable salt thereof of claim 5 or 6, wherein, one of the structures selected from Table II.

8. An antibody-drug conjugate represented by the general formula (Ab-L-IV-1), (Ab-L-IV-2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: wherein E is -O-CHR b -C(O)-, -O-(CH2) m1 -C(O)NH-, -NR d -CHR b -C(O)-, -O-(CH2) m1 -C(O)NR d -CHR b -C(O)-, Preferred -O-CHR b -C(O)-, -O-CH2-C(O)NH-, -NR d -CHR b -C(O)-、-O-CH2-C(O)NR d -CHR b -C(O)-、 R b C replaced by 1-4 Rs 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl, preferably R b Cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl groups substituted with 1-4 R groups; R d is hydrogen, deuterium, C 1-6 alkyl, preferably R d is hydrogen, deuterium, C 1-3 alkyl; or R b and R d with the atom to which it is attached forms a 4-6 membered heterocycloalkyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxy, oxo, preferably R b and R d with the carbon atom to which it is attached forms an azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl group, optionally further substituted with 1-4 R, and R is not halogen, hydroxy, oxo; R a is hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 deuteroalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxyalkyl, C 1-6 alkylamino, C 1-6 alkylaminoalkyl, C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-10 aryl or 5- to 6-membered heteroaryl, wherein said C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-10 aryl or 5- to 6-membered heteroaryl are optionally further substituted with 1 to 4 R, preferably hydrogen, deuterium, halogen, hydroxyl, cyano, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 alkoxyalkyl, C 1-3 alkylamino, C 1-3 alkylaminoalkyl, C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-8 aryl or 5- to 6-membered heteroaryl, wherein said C 3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C 6-8 aryl or 5- to 6-membered heteroaryl are optionally further substituted with 1 to 4 R; R is deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, C 1-3 haloalkylidene, C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl, wherein said C 3-6 cycloalkylidene, C 3-6 cycloalkyl, 3-6 membered heterocycloalkylidene, 3-6 membered heterocycloalkyl, C 6-10 aryl or 5-6 membered heteroaryl are optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 1-3 alkylidene, or C 1-3 haloalkylidene, preferably deuterium, halogen, hydroxyl, cyano, amino, oxo, C 1-3 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, n1 is 1 or 2; n2 is 1 or 2; m1 is 1, 2, 3; m2 is 1, 2, 3; L is a linker unit, preferably -L1-L2-L3-L4-, the L1 end is attached to Ab, and the L4 end is attached to E, wherein: L1is -(succinimid-3-yl-N)-Y-C(O)-, -CH2-C(O)-NR L1 -Y-C(O)-, -C(O)-Y-C(O)-, wherein Y is C 1-6 alkyl, C 1-6 heteroalkyl, -C 1-6 alkyl(C 3-6 cycloalkyl) or -C 1-6 heteroalkyl (3-6 membered heterocycloalkyl); said heteroalkyl containing 1-3 atoms selected from N, O or S, said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl being optionally further substituted with 1-4 groups selected from deuterium, halogen, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy; L2is a bond, -NR L2 (CH2CHO) p CH2CH2C(O)-, -NR L2 (CH2CH2O) p CH2C(O)-, -S(CH2) p C(O)-, where p is an integer from 0 to 20; L3 is a peptide residue consisting of 2-7 amino acids, wherein said amino acids are optionally further substituted with 1-4 groups selected from deuterium, halo, hydroxyl, cyano, amino, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy; L4is a bond, -NR L4 (CR L5 R L6 ) q - C(O)NR L4 (CH2) q - wherein q is an integer from 0 to 6; R L2 and R L4 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy; R L5 and R L6 each independently is hydrogen, deuterium, halogen, C 1-3 alkyl, C 1-3 deuteroalkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-6 cycloalkyl, C 3-6 cycloalkylalkyl, 3-6 membered heterocycloalkyl; Ab is an antibody, antibody fragment, or fragment linked to an antigen; z is an integer or decimal number from 1 to 10.

9. The antibody-drug conjugate, stereoisomer or pharmaceutically acceptable salt thereof according to claim 8, wherein, The L is the following structure: wherein p is an integer from 1 to 10, and the * end is attached to Ab.

10. The antibody-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to claim 8 or 9, characterized in that, one of the structures selected from Table III, wherein Ab is an antibody, antibody fragment, or fragment linked to an antigen, and z is an integer or decimal number from 1 to 10.

11. The antibody-drug conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof according to any one of claims 8-10, characterized in that, The Ab is selected from the group consisting of murine, chimeric, humanized, fully human, antibody fragments, bispecific and multispecific antibodies, preferably: an anti-EGFRvIII antibody, an anti-DLL-3 antibody, an anti-PSMA antibody, an anti-CD70 antibody, an anti-MUC16 antibody, an anti-ENPP3 antibody, an anti-TDGF1 antibody, an anti-ETBR antibody, an anti-MSLN antibody, an anti-TIM-1 antibody, an anti-LRRC15 antibody, an anti-LIV-1 antibody, an anti-CanAg / AFP antibody, an anti-cladin 18.2 antibody, an anti-Mesothelin antibody, an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-c-MET antibody, an anti-SLITRK6 antibody, an anti-KIT / CD117 antibody, an anti-STEAP1 antibody, an anti-SLAMF7 / CS1 antibody, an anti-NaPi2B / SLC34A2 antibody, an anti-GPNMB antibody, an anti-HER3 (ErbB3) antibody, an anti-MUC1 / CD227 antibody, an anti-AXL antibody, an anti-CD166 antibody, an anti-B7-H3 (CD276) antibody, an anti-PTK7 / CCK4 antibody, an anti-PRLR antibody, an anti-EFNA4 antibody, an anti-5T4 antibody, an anti-NOTCH3 antibody, an anti-Nectin 4 antibody, an anti-TROP-2 antibody, an anti-CD142 antibody, an anti-CA6 antibody, an anti-GPR20 antibody, an anti-CD174 antibody, an anti-CD71 antibody, an anti-EphA2 antibody, an anti-LYPD3 antibody, an anti-FGFR2 antibody, an anti-FGFR3 antibody, an anti-FR alpha antibody, an anti-CEACAMs antibody, an anti-GCC antibody, an anti-Integrin Av antibody, an anti-CAIX antibody, an anti-P-cadherin antibody, an anti-GD3 antibody, an anti-Cadherin 6 antibody, an anti-LAMP1 antibody, an anti-FLT3 antibody, an anti-BCMA antibody, an anti-CD79b antibody, an anti-CD19 antibody, an anti-CD33 antibody, an anti-CD56 antibody, an anti-CD74 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD37 antibody, an anti-CD47 antibody, an anti-CD138 antibody, an anti-CD352 antibody, an anti-CD25 antibody, an anti-CEACAM5 antibody, or an anti-CD123 antibody.

12. A pharmaceutical composition comprising a therapeutically effective dose of a compound of any one of claims 1-4 or an antibody-drug conjugate of any one of claims 8-10, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

13. The pharmaceutical composition of claim 12, comprising 1-1500 mg of a compound of any one of claims 1-4 or an antibody-drug conjugate of any one of claims 8-10, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

14. Use of a compound according to any one of claims 1-4 or an antibody-drug conjugate according to any one of claims 8-10, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 12 or 13 for the manufacture of a medicament, preferably a medicament for the prevention and / or treatment of a cell abnormal proliferation disease.

15. Use according to claim 14, wherein the cell abnormal proliferation disease is a tumor, preferably a solid tumor and a blood tumor, more preferably breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, glioblastoma multiforme, lymphoma, lung cancer, colon cancer, rectal cancer, colorectal cancer, leukemia, bone cancer, skin cancer, thyroid cancer, pancreatic cancer.

16. A method for treating a disease in a mammal, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-4 or an antibody-drug conjugate according to any one of claims 8-10, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 12 or 13, wherein the therapeutically effective amount is preferably 1-1500 mg, and the disease is preferably a tumor, more preferably breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, glioblastoma multiforme, lymphoma, lung cancer, colon cancer, rectal cancer, colorectal cancer, leukemia, bone cancer, skin cancer, thyroid cancer, pancreatic cancer.

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