NMT inhibitor, antibody-drug conjugate thereof, and pharmaceutical use thereof

Abstract

The present application relates to the technical field of pharmaceuticals, and specifically relates to an NMT inhibitor, an antibody-drug conjugate thereof, and pharmaceutical use thereof. In the present application, by means of structural modification, a series of structurally novel NMT inhibitors and antibody-drug conjugates thereof are obtained, and they exhibit good anti-cancer activity.

Description

An NMT inhibitor, its antibody-drug conjugate, and pharmaceutical uses

[0001] Cross-reference to related applications

[0002] This application claims priority to Chinese Patent Application No. CN202510501070.3, filed on April 18, 2025, entitled “An NMT Inhibitor and Its Antibody-Drug Conjugate and Pharmaceutical Use”, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This application relates to the field of pharmaceutical technology, specifically to an NMT inhibitor, its antibody-drug conjugate, and its pharmaceutical uses. Background Technology

[0004] Myristyltransferase (NMT) catalyzes the irreversible co-translational transfer of myristic acid from myristoyl-CoA to a protein substrate containing an N-terminal glycine. N-Myristylation is an important protein-lipid modification and co-translational modification process in eukaryotes (fungi and protists), playing a crucial role in cellular processes such as signal cascade transduction, vesicle transport, and protein localization. Higher organisms, such as humans, have two types of NMT (NMT1 and NMT2), widely expressed in various tissues, with potential substrates distributed across different cellular locations and possessing diverse biological functions. NMT is involved in the development and progression of various diseases, such as cancer, epilepsy, Alzheimer's disease, and viral, bacterial, and parasitic infections.

[0005] NMT inhibitors have been extensively studied for their application in treating microbial infections and proliferative disorders. In animal models, NMT inhibitors selectively kill cancer cells in vitro, induce regression of hematologic malignancies, and inhibit the growth of lung and breast cancer. In acute myeloid leukemia models, NMT inhibitors kill leukemia stem cell populations and promote the regeneration and growth of normal bone marrow cells. Myricx Bio has achieved complete tumor regression in multiple tumor models by using NMT inhibitors as the payload of its ADC (anti-tumor agonist) drugs.

[0006] Antibody-drug conjugates (ADCs) are therapeutic agents composed of an antibody that specifically targets a related antigen and a small molecule toxin conjugated via a linker. They combine the potent killing effect of small molecule chemotherapeutic agents with the tumor-targeting properties of antibody drugs. After entering the body, ADC molecules bind to antigens on the surface of target cells through antibody guidance, releasing toxin molecules via chemical or enzymatic reactions to destroy the target cells. Due to their targeting and intracellular drug release, ADCs can precisely bind to tumors and reduce the impact on normal cells, increasing local drug concentration, reducing toxic side effects on normal cells, and expanding the therapeutic window. ADCs have broad application prospects in cancer treatment and have achieved some encouraging results in clinical research.

[0007] Currently, there are no NMT inhibitor drugs on the market. Therefore, it is still necessary to develop new compounds with better efficacy, pharmacokinetic behavior, and reduced toxicity, or to use them as payloads for ADC molecules to improve efficacy and safety in vivo. Summary of the Invention

[0008] In order to achieve the purpose of this invention, this application provides an NMT inhibitor and its antibody-drug conjugate and pharmaceutical use. The NMT inhibitor and its antibody-drug conjugate described in this application have good anticancer activity.

[0009] In a first aspect, this application provides an NMT inhibitor, which is a compound of formula (I), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt thereof.

[0010] in:

[0011] R1 is a group of formula -XLA;

[0012] L represents -(CR9R) 10 )r-3-8-membered cycloalkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered cycloalkylene-O-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-O-*、-(CR9R 10 )tO-* or -(CR9R 10 )t-(NR 11)s*; * represents the linking site with group A, r is 0, 1, 2, 3 or 4, s is 0 or 1, t is 1, 2, 3, 4, 5 or 6; each R9, R 10 Each of these can be independently H, halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, -COOH, or -CON(R). 11 2. 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; R 11 H, optionally substituted C1-8 alkyl, -COOH, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl;

[0013] The cycloalkylene, heteroalkylene, cycloalkylene, and heteroalkylene groups may optionally be substituted with substituents selected from the following: halogen, amino, hydroxyl, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino.

[0014] X represents -O-, -NR 11 -, -S- or a single key;

[0015] A is a 6-10 aryl or a 5-10 heteroaryl group, wherein the aryl or heteroaryl group is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino, optionally substituted C1-8 alkylacyl, -COOH, -CON(R) 11 )2;

[0016] Y is C or N (carbon or nitrogen);

[0017] Each R2 is independently a halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio or optionally substituted C1-8 alkylamino; m is 0, 1, 2 or 3 or 4.

[0018] K, Z, and M are each independently C or N (carbon or nitrogen);

[0019] Each R3 is independently a halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, or optionally substituted C1-8 alkylamino; n is 0, 1, 2, or 3.

[0020] Each R4 is independently H or an optional substituted C1-8 alkyl group, and p is 1, 2 or 3;

[0021] Each R5 and R6 is independently H, halogen, hydroxyl, amino, nitro, cyano, mercapto, or optionally substituted C1-8 alkyl, and q is 1, 2, 3, or 4.

[0022] R7 and R8 are each independently H or optionally substituted C1-8 alkyl groups;

[0023] The optional substituents in the C1-8 alkyl, C1-8 alkoxy, C1-8 alkylthio, C1-8 alkylamino, and C1-8 alkylacyl groups are selected from the following groups: halogen, hydroxyl, amino, nitro, cyano, mercapto, C1-8 alkoxy, C1-8 alkylamino, C1-8 alkylacyl, -COOH, -CON(R) 11 2, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl.

[0024] Specifically, the optional substitution may refer to substitution by 0, 1, 2 or 3 substituents.

[0025] In one set of implementations, X stands for -O-.

[0026] For L groups:

[0027] In one set of implementations, L stands for -(CR9R) 10 )r-3-8-membered cycloalkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered cycloalkylene-O-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-O-*、-(CR9R 10 )tO-* or -(CR9R 10 )t-(NR 11 )s-.

[0028] In one set of implementations, L represents -(CR9R) 10 )r-3-6-membered cycloalkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-6-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-6-membered cycloalkylene-O-*、-(CR9R 10 )r-3-6-membered heterocyclic alkylene-O-* or -(CR9R 10 )t-(NR 11 )s-*; r is 0, 1 or 2, s is 0 or 1, t is 1, 2 or 3.

[0029] In one set of implementations, each R9, R 10 Each of these can be independently H, C1-8 alkyl, hydroxyl-substituted C1-8 alkyl, amino-substituted C1-8 alkyl, C1-8 alkoxy-substituted C1-8 alkyl, C1-8 alkylamino-substituted C1-8 alkyl, -COOH-substituted C1-8 alkyl, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl; R 11 It can be H, C1-8 alkyl, hydroxyl-substituted C1-8 alkyl, amino-substituted C1-8 alkyl, C1-8 alkoxy-substituted C1-8 alkyl, C1-8 alkylamino-substituted C1-8 alkyl, -COOH-substituted C1-8 alkyl, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl.

[0030] In one set of embodiments, L represents a 3-6 membered cyclohexene alkyl group, -(CR9R) 10 )r-3-6-membered cycloalkylene*, -3-6-membered cycloalkylene-NR 11 -* or -(CR9R) 10 )r-3-6-membered cycloalkylene-NR 11 -*; r is 1 or 2. Preferably, R9, R 10 R 11 All are H.

[0031] In one set of implementations, L represents -(CR9R) 10 )t-NR 11 -* or -(CR9R) 10 )t-, where t is 1, 2, or 3, optionally, R9, R 10 R 11 At least one (preferably one) is a hydroxyl-substituted C1-8 alkyl, an amino-substituted C1-8 alkyl, a C1-8 alkoxy-substituted C1-8 alkyl, a C1-8 alkylamino-substituted C1-8 alkyl, a -COOH-substituted C1-8 alkyl, a 3-6 membered cycloalkyl, or a 3-6 membered heterocycloalkyl. Examples include hydroxymethyl, hydroxyethyl, alkoxymethyl, alkoxyethyl, alkylaminomethyl, alkylaminoethyl, -COOH-substituted methyl, and 3-epoxybutyl. Optionally, R9, R 10 R 11 One of them is a hydroxyl-substituted C1-8 alkyl group. Preferably, R9, R 10 R 11 Each can be independently H, methyl, ethyl, n-propyl, n-butyl, hydroxymethyl, or hydroxyethyl.

[0032] In one set of implementations, L represents -(CR9R) 10 )t-NR 11 -*, t is 1, 2, 3, 4, 5 or 6, R9, R 10 R 11Selected from H and C1-8 alkyl groups. Preferably, L represents -(CH2)t-NR. 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups. More preferably, L represents -(CH2)2-NR. 11 *, R 11 It can be H, methyl, ethyl, n-propyl or n-butyl.

[0033] In one set of implementations, L represents -CH2CH2-NH-*, -CH2CH2-N(-CH3)-*, -CH2CH2-N(-CH2CH3)-*, -CH2CH2-N(-CH2CH2CH3)-*, -CH2CH2-N(-CH2CH2CH2CH3)-*, -CH(-CH2OH)CH2-N(-CH3)-*, -CH2CH(-CH2OH)-N(-CH3)-*, -CH2CH2-N(-CH2CH2OH)-*, -CH2CH2-* or -CH2CH2-O-*; * represents the connection site with group A. Preferably, L represents -CH2CH2-NH-*, -CH2CH2-N(-CH3)-*, -CH2CH2-N(-CH2CH3)-*, -CH2CH2-N(-CH2CH2CH3)-*, or -CH2CH2-N(-CH2CH2CH2CH3)-*; * represents the linking site with group A.

[0034] For group A:

[0035] In one set of embodiments, A is a 6-10 aryl or a 5-10 heteroaryl, wherein the aryl or heteroaryl is optionally substituted with a substituent selected from the following: C1-8 alkyl, C1-8 alkylacyl, -COOH, -CO(N 11 )2, and those affected by hydroxyl, C1-8 alkyl acyl, -COOH, -CON(R) 11 2. C1-8 alkyl groups substituted with 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl. Preferably, the aryl or heteroaryl group is optionally substituted with a substituent selected from the following: C1-8 alkyl, C1-8 alkyl acyl, or C1-8 alkyl group substituted with a hydroxyl group. More preferably, the aryl or heteroaryl group is optionally substituted with a substituent selected from the following: C1-4 alkyl, C1-4 alkyl acyl, or C1-5 alkyl group substituted with a hydroxyl group.

[0036] In one set of embodiments, A is a 5-6-membered heteroaryl group containing 1, 2, or 3 nitrogen atoms. Preferably, A is a 4-pyrazolyl group.

[0037] In one set of implementations, A is

[0038] Among them, R 12 R 13 Each is independently selected from H, halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino or optionally substituted C1-8 alkylacyl.

[0039] R 14 R 15 R 16 R 17 Each is independently selected from H, halogen, hydroxyl, amino, nitro, cyano, mercapto, or optionally substituted C1-8 alkyl groups;

[0040] v is 0, 1, or 2;

[0041] The substituents in the optionally substituted C1-8 alkyl, C1-8 alkoxy, C1-8 alkylthio, C1-8 alkylamino, and C1-8 alkylacyl groups are selected from the following groups: halogen, hydroxyl, amino, nitro, cyano, and mercapto.

[0042] In one set of implementations, A is L represents -(CR9R) 10 )r-3-8-membered cycloalkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered cycloalkylene-O-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-O-*、-(CR9R 10 )tO-* or -(CR9R 10 )t-NR 11 -

[0043] In one set of implementations, A is L represents -(CR9R) 10 )r-3-8-membered cycloalkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered cycloalkylene-O-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-O-*、-(CR9R 10 )tO-* or -(CR9R 10 )t-(NR 11)s-*.

[0044] In one set of implementations, R 12 R 13 Each is independently selected from H, C1-8 alkyl; R 14 R 15 R 16 Each is independently selected from H, C1-8 alkyl; R 17 Selected from C1-8 alkyl groups; v is 0, 1 or 2.

[0045] In one set of implementations, R 12 R 13 Each is independently selected from C1-8 alkyl groups; R 16 Selected from H;R 17 Selected from C1-8 alkyl groups; v is 0. Preferably, R 12 R 13 Each is independently selected from methyl groups. Preferably, R 17 Selected from isopropyl or tert-butyl.

[0046] In one set of implementations, A is R 12 R 13 Each is independently selected from H, C1-8 alkyl; R 14 R 15 R 16 Each is independently selected from H, C1-8 alkyl; R 17 Selected from C1-8 alkyl groups; v is 0, 1, or 2; L represents -(CR9R) 10 )t-NR 11 -*, t is 1, 2, 3, 4, 5 or 6, R9, R 10 R 11 Selected from H and C1-8 alkyl groups.

[0047] Preferably, R 12 R 13 Each is independently selected from C1-4 alkyl groups; R 16 Selected from H;R 17 Selected from C1-4 alkyl groups; v is 0; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H, C1-4 alkyl. More preferably, R 12 R 13 Each is independently selected from methyl, R 17 Selected from isopropyl or tert-butyl, L represents -(CH2)2-NR 11 *, R 11 It is H, methyl, ethyl, n-propyl, or n-butyl. Most preferably, R... 12 R 13Each is independently selected from methyl, R 17 Selected from isopropyl or tert-butyl, L represents -(CH2)2-NR 11 *, R 11 It can be methyl or ethyl.

[0048] In one set of embodiments, each R2 is independently a halogen; m is 0, 1, or 2. Preferably, each R2 is independently F or Cl; m is 2.

[0049] In one set of embodiments, K and Z are N, and M is C; or K, Z, and M are all N; or K is C, and Z and M are N. Preferably, K and Z are N, and M is C.

[0050] In one set of implementations, n is 0.

[0051] In one set of implementations, each R4 is independently H.

[0052] In one set of implementations, each R5 and R6 is independently H.

[0053] In one set of implementations, q is 1 or 2.

[0054] In one set of embodiments, R7 and R8 are each independently H or C1-8 alkyl; preferably, R7 and R8 are H or methyl, respectively.

[0055] In one set of embodiments, R1 is a group of formula -XLA;

[0056] L represents -(CR9R) 10 )t-NR 11 * represents the binding site with group A, where t is 1, 2, 3, 4, 5, or 6; R9, R 10 R 11 Selected from H, C1-8 alkyl;

[0057] X stands for -O-;

[0058] A is a 5-10 membered heteroaryl group, wherein the heteroaryl group is optionally substituted with a substituent selected from the following: C1-8 alkyl groups optionally substituted with a hydroxyl group;

[0059] Y is C or N (carbon or nitrogen);

[0060] Each R2 is independently a halogen; m is 0, 1, 2, 3, or 4;

[0061] K, Z, and M are each independently C or N (carbon or nitrogen);

[0062] n is 0, 1, 2, or 3;

[0063] Each R4 is independently H or an optional substituted C1-8 alkyl group, and p is 1, 2 or 3;

[0064] Each R5 and R6 is independently H, and q is 1, 2, 3 or 4;

[0065] R7 and R8 are each independently H or optionally substituted C1-8 alkyl groups.

[0066] Preferably, L represents -(CH2)t-NR 11 -*; * represents the binding site with group A, t is 1, 2 or 3; R 11 It is an H or C1-4 alkyl group;

[0067] A is a 5-10 membered heteroaryl group, wherein the heteroaryl group is optionally substituted by a substituent selected from the following: C1-4 alkyl, C1-5 alkyl substituted with hydroxyl;

[0068] Y is C;

[0069] Each R2 is independently a halogen; m is 2;

[0070] K and Z are each independently N, and M is C;

[0071] n is 0;

[0072] Each R4 is independently H, and p is 1;

[0073] Each R5 and R6 is independently H, and q is 1;

[0074] R7 and R8 are each independently H or optionally substituted C1-4 alkyl groups.

[0075] More preferably, L represents -(CH2)2-NR 11 -*; * represents the connection site with group A, R 11 It can be H, methyl, ethyl, n-propyl, or n-butyl;

[0076] A is a 5-10-membered heteroaryl group, wherein the heteroaryl group is optionally substituted by a substituent selected from the following: C4-5 alkyl groups substituted with methyl or hydroxyl groups;

[0077] R7 and R8 are H or methyl, respectively.

[0078] In one set of embodiments, the compound of formula (I) is either the compound of formula (I-1) or the compound of formula (I-2):

[0079] The definitions of each group are as described above.

[0080] In one set of embodiments, the compound of formula (I) is a compound of formula (I-3) or a compound of formula (I-4):

[0081] The definitions of each group are as described above.

[0082] Preferably, each R2 is independently F or Cl. 16 Selected from H, R 17 Selected from isopropyl or tert-butyl. R7 and R8 are each independently H or C1-8 alkyl.

[0083] More preferably, each R2 is independently F, and R7 and R8 are each independently H or methyl; L represents -(CR9R) 10 )t-NR 11 -*, t is 1, 2, 3, 4, 5 or 6, R9, R 10 R 11 Selected from H and C1-8 alkyl groups.

[0084] Most preferably, R7 and R8 are H or methyl groups, respectively; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups.

[0085] Most preferably, the compound of formula (I) is a compound of formula (I-3), R 16 Selected from H, R 17 Selected from isopropyl or tert-butyl; R7 and R8 are H or methyl, respectively; L represents -(CH2)2-NR 11 *, R 11 The form is H, methyl, ethyl, n-propyl, or n-butyl. Most preferably, the compound of formula (I) is a compound of formula (I-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be methyl or ethyl.

[0086] In one set of embodiments, the compound of formula (I) is one of the following compounds:

[0087] In a second aspect, this application provides an NMT inhibitor, which is a compound of formula (II), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt thereof.

[0088] in:

[0089] Q represents a linker group that can be conjugated to antibody or antigen binding fragments;

[0090] L1 represents the linking group;

[0091] Other groups are defined as described in the first aspect.

[0092] In one set of embodiments, Q represents a linker group comprising a maleimide structure, a linker group comprising a methanesulfonylpyrimidine structure, a linker group comprising a methanesulfonylpyridinothiazole structure, a linker group comprising a methanesulfonylpyrimidineothiazole structure, a linker group comprising a cycloenyne structure, or a linker group comprising an oxime bond structure. Preferably, Q represents a linker group comprising a maleimide structure or a linker group comprising a methanesulfonylpyridinothiazole structure.

[0093] In one set of implementations, Q-L1- represents

[0094] In one set of embodiments, the compound of formula (II) is a compound of formula (II-1) or a compound of formula (II-2).

[0095] in:

[0096] Q represents a linker group that can be conjugated to antibody or antigen binding fragments;

[0097] L1 represents the linking group;

[0098] Other groups are defined as described in the first aspect.

[0099] In one set of embodiments, the compound of formula (II) is a compound of formula (II-3) or a compound of formula (II-4).

[0100] in:

[0101] Q represents a linker group that can be conjugated to antibody or antigen binding fragments;

[0102] L1 represents the linking group;

[0103] Other groups are as described in the first aspect. Preferably, each R2 is independently F or Cl, R 16 Selected from H, R 17 It is selected from isopropyl or tert-butyl, and R7 and R8 are each independently H or C1-8 alkyl.

[0104] More preferably, each R2 is independently F, and R7 and R8 are each independently H or methyl; L represents -(CR9R) 10 )t-NR 11 -*, t is 1, 2, 3, 4, 5 or 6, R9, R 10 R 11 Selected from H and C1-8 alkyl groups.

[0105] Most preferably, R7 and R8 are H or methyl groups, respectively; L represents -(CH2)t-NR 11-*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups.

[0106] Most preferably, the compound of formula (II) is of formula (II-3), and compound R 16 Selected from H, R 17 Selected from isopropyl or tert-butyl; R7 and R8 are H or methyl, respectively; L represents -(CH2)2-NR 11 *, R 11 The form is H, methyl, ethyl, n-propyl, or n-butyl. Most preferably, the compound of formula (II) is a compound of formula (II-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be methyl or ethyl.

[0107] In one set of embodiments, the compound of formula (II) is the following compound:

[0108] In a third aspect, this application provides an antibody-drug conjugate formed by conjugating the NMT inhibitor described in the second aspect to an antibody or antigen-binding fragment Ab via a linker group.

[0109] In one set of embodiments, the antibody-drug conjugate is a compound of formula (III), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt thereof.

[0110] in:

[0111] Ab represents an antibody or antigen-binding fragment;

[0112] L' is a direct bond or a linking group connecting Ab and Q';

[0113] Q' is the group formed by connecting Q to Ab through L';

[0114] w is an integer or decimal between 0 and 20; such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 0 and decimals between the above integers;

[0115] Other groups are defined as described in the second aspect.

[0116] In one set of embodiments, Q' represents a linker group comprising a maleimide structure, a linker group comprising a methanesulfonylpyrimidine structure, a linker group comprising a methanesulfonylpyridinothiazole structure, a linker group comprising a methanesulfonylpyrimidineothiazole structure, a linker group comprising a cycloeneyne structure, or a linker group comprising an oxime bond structure.

[0117] In one set of implementations, -Q'-L1- represents

[0118] Maleimide or pyridothiazole is connected to L' at one end.

[0119] In one set of implementations, w is an integer or decimal between 0 and 8.

[0120] In one set of embodiments, Ab is selected from murine antibodies, chimeric antibodies, humanized antibodies, fully human antibodies, or antigen-binding fragments thereof.

[0121] In one set of implementations, Ab is selected from monoclonal antibodies and polyclonal antibodies (such as bispecific antibodies and triple antibodies). Preferably, the Ab is selected from anti-HER2 (ErbB2) antibody, anti-EGFR antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-ROR1 antibody, anti-CLDN6 antibody, anti-CLDN9 antibody, anti-CLDN18.2 antibody, anti-NaPi-2b antibody, anti-TNF-α antibody, anti-ENPP3 antibody, anti-DLL3 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD28 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD38 antibody, anti-CD44 antibody, anti-CD45 antibody, anti-CD47 antibody, anti-CD48 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD98 antibody, anti-CD105 antibody, anti-CEA antibody, anti-EphA2 antibody, anti-MUCI antibody, and anti-Lewis antibody. Anti-γ antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-CD79 antibody, anti-TROP-2 antibody, anti-CD79B antibody, anti-Mesothelin antibody, anti-Nectin-4 antibody, anti-TPBG antibody, or their antigen-binding fragments.

[0122] Preferably, Ab is an anti-HER2 (ErbB2) antibody or its antigen-binding fragment.

[0123] Preferably, Ab is selected from monoclonal antibodies or their antigen-binding fragments, and more preferably from trastuzumab, cetuximab, pertuzumab, nimotuzumab, enoblituzumab, emibetuzumab, Inotuzumab, pinatuzumab, brentuximab, gemtuzumab, bivatuzumab, lorvotuzumab, or their antigen-binding fragments; more preferably, Ab is selected from trastuzumab or its antigen-binding fragments.

[0124] In one set of embodiments, the compound of formula (III) is a compound of formula (III-1) or a compound of formula (III-2).

[0125] in:

[0126] Ab represents an antibody or antigen-binding fragment;

[0127] L' is a direct bond or a linking group connecting Ab and Q';

[0128] Q' is the group formed by connecting Q to Ab through L';

[0129] w is an integer or decimal between 0 and 20; such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 0 and decimals between the above integers;

[0130] Other groups are defined as described in the second aspect.

[0131] In one set of embodiments, the compound of formula (III) is a compound of formula (III-3) or a compound of formula (III-4).

[0132] in:

[0133] Ab represents an antibody or antigen-binding fragment;

[0134] L' is a direct bond or a linking group connecting Ab and Q';

[0135] Q' is the group formed by connecting Q to Ab through L';

[0136] w is an integer or decimal between 0 and 20; such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 0 and decimals between the above integers;

[0137] Other groups are defined as described in the second aspect.

[0138] Preferably, each R2 is independently F or Cl, R 16 Selected from H, R 17 It is selected from isopropyl or tert-butyl, and R7 and R8 are each independently H or C1-8 alkyl.

[0139] More preferably, each R2 is independently F, and R7 and R8 are each independently H or methyl; L represents -(CR9R) 10 )t-NR 11 -*, t is 1, 2, 3, 4, 5 or 6, R9, R 10 R 11 Selected from H and C1-8 alkyl groups.

[0140] Most preferably, R7 and R8 are H or methyl groups, respectively; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups.

[0141] Most preferably, the compound of formula (III) is a compound of formula (III-3), R 16 Selected from H, R 17 Selected from isopropyl or tert-butyl; R7 and R8 are H or methyl, respectively; L represents -(CH2)2-NR 11 *, R 11 The form is H, methyl, ethyl, n-propyl, or n-butyl. Most preferably, the compound of formula (III) is a compound of formula (III-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be methyl or ethyl.

[0142] In one set of embodiments, the compound of formula (III) is the following compound:

[0143] Ab and w are defined as described above. Preferably, Ab is an anti-HER2 (ErbB2) antibody or its antigen-binding fragment, more preferably trastuzumab or its antigen-binding fragment.

[0144] In a fourth aspect, this application provides a pharmaceutical composition comprising the NMT inhibitor described in the first and second aspects of this application, the antibody-drug conjugate described in the third aspect, and one or more pharmaceutically acceptable carriers or excipients.

[0145] In a fifth aspect, this application provides the use of the NMT inhibitors described in the first and second aspects of this application in the preparation of antibody-drug conjugates.

[0146] In a sixth aspect, this application provides the use of the NMT inhibitors described in the first and second aspects of this application, the antibody-drug conjugates described in the third aspect, and the pharmaceutical compositions described in the fourth aspect in the preparation of a medicament for the prevention or treatment of tumors or cancer; or the use in the prevention or treatment of tumors or cancer.

[0147] In a seventh aspect, this application provides a method for preventing or treating tumors or cancer, comprising administering to a subject in need an effective amount of the NMT inhibitor according to the first and second aspects of this application, the antibody-drug conjugate according to the third aspect, and the pharmaceutical composition according to the fourth aspect.

[0148] In one set of embodiments, the tumor or cancer is selected from solid tumors or hematologic malignancies; preferably, the tumor or cancer is selected from metastatic or recurrent tumors or cancers; preferably, the tumor or cancer is selected from breast cancer, gastric cancer, colon cancer, pancreatic cancer, kidney cancer, ovarian cancer, lung cancer, soft tissue sarcoma, liposarcoma, melanoma, head and neck cancer, cervical cancer, and prostate cancer; specifically, it may be HER-2 positive breast cancer, triple-negative breast cancer, HER-2 positive triple-negative breast cancer, renal clear glandular adenocarcinoma, small cell lung cancer, non-small cell lung cancer, or large cell lung cancer.

[0149] In one set of embodiments, the tumor or cancer is selected from cancers or tumors that highly express HER2 (ErbB2) antibodies.

[0150] This application obtains a series of novel NMT inhibitors and their antibody-drug conjugates through structural modification, which have good anticancer activity. Attached Figure Description

[0151] Figure 1 shows the tumor volume change curve of the JIMT-1 breast cancer-bearing mouse model. Detailed Implementation

[0152] Unless otherwise stated, all terms used in disclosing this application (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains, and the following definitions are provided for a better understanding of the teachings of this application. The explanations of the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting of this application.

[0153] Features described or illustrated as part of one set of embodiments in this application may be used in another set of embodiments to produce further embodiments.

[0154] The terms "and / or," "or / and," and "and / or" as used in this application encompass any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected using at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes solutions connected by "logical AND," and also undoubtedly includes solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0155] The terms “containing,” “comprising,” and “including” as used in this application are synonyms and are inclusive or open-ended, and do not exclude additional, uncited members, elements, or method steps.

[0156] The range of values ​​represented by endpoints in this application includes all values ​​and fractions contained within that range, as well as the endpoints referenced.

[0157] In this application, concentration values ​​are defined as those within a certain range of fluctuation. For example, fluctuations are allowed within a corresponding precision range. For instance, 2% may fluctuate within ±0.1%. For larger values ​​or values ​​that do not require extremely fine control, even greater fluctuations are permitted. For example, 100mM may fluctuate within ranges of ±1%, ±2%, ±5%, etc. Regarding molecular weight, fluctuations of ±10% are allowed.

[0158] In this application, the terms "multiple" or "various" are used unless otherwise specified, referring to a quantity of 2 or more.

[0159] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0160] In this application, terms such as "preferred," "better," "more suitable," "specific," and "preferably" are merely descriptions of more effective implementation methods or embodiments, and should be understood not to limit the scope of protection of this application. In this application, terms such as "optional," "optionally," "optional," and "optional" mean that something is optional, that is, selected from either "with" or "without" parallel solutions. If multiple "optional" options appear in a technical solution, unless otherwise specified and without contradiction or mutual constraint, each "optional" option is independent.

[0161] The term "isotope-labeled compound" refers to an isotope-labeled compound obtained by replacing any one atom in a compound with its isotopic atom. All pharmaceutically acceptable isotope-labeled compounds described in this application include those in which one or more atoms are replaced by atoms having the same atomic number as atoms commonly found in nature but with different atomic masses or mass numbers. Examples of isotopes suitable for inclusion in the compounds of this application include isotopes of hydrogen, such as... 2 H(D) and 3 H(T), isotopes of carbon, such as 11 C 13 C and 14 C, isotopes of chlorine, such as 37 Cl, an isotope of fluorine, such as 18 F, an isotope of iodine, such as 123 I and 125 I, isotopes of nitrogen, such as 13 N and 15 N, an isotope of oxygen, such as 15 O、 17 O and 18 O, and isotopes of sulfur, such as 35 S. Isotope-labeled compounds can generally be prepared using conventional techniques known to those skilled in the art, or by using a suitable isotope-labeling reagent instead of the previously used unlabeled reagent, in a manner similar to that described in the examples and preparations appended herein.

[0162] The term "optical isomer" refers to the various isomers formed when a compound has one or more chiral centers, each of which can exist in either an R or S configuration. Optical isomers include all diastereomers, enantiomers, meso compounds, racemates, or mixtures thereof. For example, optical isomers can be separated by chiral chromatography or by chiral synthesis.

[0163] The term "geometric isomer" refers to the fact that when a compound contains a double bond, it can exist as a cis isomer, a trans isomer, an E-isomer, or a Z-isomer. Geometric isomers include cis isomers, trans isomers, E-isomers, Z-isomers, or mixtures thereof.

[0164] The term "tautomer" refers to an isomer that results from the rapid movement of an atom in a molecule to two different positions. Those skilled in the art will understand that tautomers can interconvert and may coexist in an equilibrium state under certain conditions.

[0165] The compounds of this application may exist in the form of pharmaceutically acceptable salts, such as acid addition salts and / or base addition salts of compounds of formula (I). Unless otherwise specified, "pharmaceutically acceptable salt" as used herein includes acid addition salts or base addition salts that may appear in the compounds of this application. Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include, but are not limited to: acetates, adipates, aspartates, benzoates, benzenesulfonates, bicarbonates / carbonates, hydrogen sulfates / sulfates, borates, camphor sulfonates, citrates, cyclohexylamine sulfonates, ethanedisulfonates, formates, fumarates, gluconate, glucuronide, glucuronide, hexafluorophosphate, 2-(4-hydroxybenzyl)benzoate, hydrochlorides / chlorides, hydrobromides / bromines, hydroiodides / iodides, 2-hydroxyethanesulfonates, lactates, malates, maleates, malonates, methanesulfonates, sulfates, methyl sulfates, naphthalates, 2-naphthalenesulfonates, nicotinates, nitrates, orotates, oxalates, hexadecates, phosphates / hydrogen phosphates / dihydrogen phosphates, pyroglutamates, gluconate, stearates, salicylates, tannins, tartrates, toluenesulfonates, and trifluoroacetates. Suitable base addition salts are formed by bases that form non-toxic salts. Examples include, but are not limited to: aluminum, arginine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc salts. They can also form acidic and basic hemisalts, such as hemisulfates and hemicalcium salts. For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection and Use by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds described in this application are known to those skilled in the art.

[0166] Some compounds of this application may exist in different crystal forms or amorphous forms, and all such compounds are included within the scope of this application, regardless of their form.

[0167] The term "antibody drug conjugate" (ADC) refers to a biologically active toxic drug, such as an NMT inhibitor, conjugated to an antibody or antigen-binding fragment via a stable linker group.

[0168] The term "linker group" refers to a chemical structural fragment or bond that is linked to an antibody at one end and to a toxin drug at the other end. It can be linked to other linker groups before being linked to an antibody (or antigen-binding fragment) or a toxin drug.

[0169] The term "antibody drug conjugate" (ADC) refers to an antibody or antigen-binding fragment linked to a biologically active toxic drug, such as an NMT inhibitor, via a stable linker unit.

[0170] In this application, the term "antibody" refers to immunoglobulin, which is a tetrapeptide chain structure composed of two identical heavy chains and two identical light chains linked by interchain disulfide bonds. The amino acid composition and sequence of the constant region of the immunoglobulin heavy chain differ, thus their antigenicity also differs. Based on this, immunoglobulins can be divided into five classes, or isotypes of immunoglobulins: IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε chains, respectively. Within the same class of Ig, based on differences in the amino acid composition of its hinge region and the number and position of disulfide bonds in the heavy chain, different subclasses can be further divided; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. The light chains are classified as κ or λ chains based on differences in the constant region. Each of the five classes of Ig can have either a κ or λ chain. The sequence of approximately 110 amino acids near the N-terminus of the antibody heavy and light chains varies considerably, forming the variable region (Fv region); the remaining amino acid sequence near the C-terminus is relatively stable, forming the constant region. The variable region comprises three hypervariable regions (HVRs) and four relatively conserved backbone regions (FRs). The three hypervariable regions determine the antibody's specificity and are also known as complementarity-determining regions (CDRs). Each light chain variable region (LCVR) and heavy chain variable region (HCVR) consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDRs of the light chain refer to LCDR1, LCDR2, and LCDR3; the three CDRs of the heavy chain refer to HCDR1, HCDR2, and HCDR3. The CDR amino acid residues in the LCVR and HCVR regions of the antibody or antigen-binding fragment described in this application conform to the known Kabat numbering rules (LCDR1-3, HCDR2-3) in number and position, or conform to the Kabat and Chothia numbering rules (HCDR1).

[0171] The term "antigen-binding fragment" refers to one or more fragments of an antibody that maintain the ability to specifically bind to an antigen. It has been shown that fragments of full-length antibodies can be used for antigen-binding function. Examples of binding fragments included in "antigen-binding fragments" include: (i) Fab fragments, monovalent fragments consisting of VL, VH, CL, and CH domains; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by disulfide bridges on the chain region; (iii) F fragments consisting of VH and CH domains; (iv) Fv fragments consisting of VH and VL domains of a single arm of the antibody; (v) single-domain or dAb fragments consisting of a VH domain; and (vi) separate complementarity-determining regions (CDRs) or (vii) combinations of two or more separate CDRs optionally linked by synthetic linkers. Furthermore, although the two domains VL and VH of the Fv fragment are encoded by separate genes, they can be linked by synthetic linkers using recombinant methods, thereby enabling the production of a single protein chain (called a single-chain Fv (scFv)) in which the VL and VH regions pair to form a monovalent molecule. Such single-chain antibodies are also included in the term "antigen-binding fragment." Such antibody fragments are obtained using conventional techniques known to those skilled in the art, and fragments are screened for functionality in the same manner as for intact antibodies. Antigen-binding moieties can be generated by recombinant DNA technology or by enzymatic or chemical cleavage of intact immunoglobulins. Antibodies can be different types of antibodies, such as IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtypes), IgA1, IgA2, IgD, IgE, or IgM antibodies.

[0172] In this application, some of the substituents are... The symbol indicates a connection site, and "*" represents a connection site.

[0173] Unless otherwise stated, "optional substitution" means that the hydrogen on the substituted group is not substituted or that the hydrogen at one or more substituted sites of the substituted group is independently substituted by a substituent. The substituent is independently selected from one or more (e.g., 1, 2, 3, or 4) of the following groups: deuterium, oxo, halogen, amino, hydroxyl, nitro, cyano, mercapto, optionally substituted alkyl, optionally substituted alkyloxy, optionally substituted alkylthio, optionally substituted alkylamino, optionally substituted alkylacyl, optionally substituted carbocyclic, and optionally substituted heterocyclic. When a substituent is selected as "oxo," it means that two hydrogen atoms on the carbon at the same substitution position are replaced by an oxygen atom, or a heteroatom is bonded to oxygen, such as nitrogen oxides bonded to oxygen, sulfinyl groups bonded to oxygen, or sulfonyl groups bonded to oxygen. "Optional oxo" means that the hydrogen on the substituted group is not substituted or that two hydrogen atoms on the carbon at the same substitution position are replaced by an oxygen atom, or a heteroatom is bonded to oxygen, such as nitrogen oxides bonded to oxygen, sulfinyl groups bonded to oxygen, or sulfonyl groups bonded to oxygen. The term "independently" means that when there are more than one substituent, these substituents can be the same or different.

[0174] The term "oxygen bond" (=O) refers to the situation where two hydrogen atoms on a carbon atom bonded to it are replaced by oxygen atoms to form a carbonyl group.

[0175] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight-chain or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms (the number of carbon atoms is between 1 and 12, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12), more preferably an alkyl group containing 1 to 10 carbon atoms, most preferably an alkyl group containing 1 to 8 carbon atoms (i.e. C1-8 alkyl, the number of carbon atoms is between 1 and 8, specifically 1, 2, 3, 4, 5, 6, 7 or 8), and an alkyl group containing 1 to 6 carbon atoms (i.e. C1-6 alkyl, the number of carbon atoms is between 1 and 6, specifically 1, 2, 3, 4, 5 or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, 3-hexyl, 2-pentyl, etc.

[0176] The term "alkyloxy" refers to -O-alkyl, where the alkyl group is defined as above. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, etc. The term "alkylthio" refers to -S-alkyl, where the alkyl group is defined as above. The term "alkylamino" refers to -NH-alkyl or -N(alkyl)-alkyl, where the alkyl group is defined as above. The term "alkylacyl" refers to -CO-alkyl, where the alkyl group is defined as above.

[0177] The term "halogen" refers to F, Cl, Br, and I.

[0178] The terms "hydroxyl-substituted alkyl" and "hydroxyl-substituted C1-8 alkyl" refer to alkyl or C1-8 alkyl groups that are substituted by one or more hydroxyl groups.

[0179] The term "cycloalkylene" refers to a divalent saturated monocyclic or polycyclic (including bicyclic, tricyclic, etc.) cyclic hydrocarbon substituent, including monocyclic, fused, spirocyclic, and bridged rings. Cycloalkylene groups may contain 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20) carbon atoms, preferably 3 to 15 carbon atoms, more preferably 3 to 12 carbon atoms, 3 to 8 (e.g., 3, 4, 5, 6, 7, 8) carbon atoms, or 3 to 6 carbon atoms. Non-limiting examples of cycloalkylene compounds include cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, cyclooctylene, 1,2-cyclopropylene, 1,2-cyclobutylene, 1,3-cyclobutylene, 1,2-cyclopentylene, 1,3-cyclopentylene, 1,3-cyclopentylene, etc.

[0180] The term "heterocyclic alkylene" refers to a divalent saturated monocyclic or polycyclic (including bicyclic, tricyclic, etc.) cyclic hydrocarbon substituent, including monocyclic, fused, combined, spirocyclic, and bridged rings, containing 3 to 20 ring atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20), wherein one or more ring atoms (e.g., 1, 2, 3, 4) are heteroatoms selected from N, O, or S. Preferably, it contains 3 to 12 ring atoms (including 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms), 3 to 8 (e.g., 3, 4, 5, 6, 7, 8) ring atoms, or 3 to 6 ring atoms, wherein 1 to 3 are heteroatoms (1, 2, or 3 heteroatoms), and the heteroatoms are selected from N, O, or S. Heterocyclic alkyl groups can be selected from 3-11 member monocyclic, fused, bicyclic, spirocyclic, and bridged heterocyclic alkyl groups with one or two nitrogen atoms, such as 3-6 member monocyclic heterocyclic alkyl groups, 6-12 member fused heterocyclic alkyl groups, 6-12 member bicyclic heterocyclic alkyl groups, 6-12 member spirocyclic heterocyclic alkyl groups, and 6-12 member bridged heterocyclic alkyl groups. Non-limiting examples include pyrrolidinyl, imidazoalkyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, aziridine, oxadiazinyl, and sulfur. Heterocyclic propane, azahexacyclic butane, oxacyclobutane, thiohexacyclic butane, oxacyclohexane, morpholino, thiomorpholino, dioxane, dithiohexacyclohexyl, oxazolyl, thiazolyl, pyrazolyl, imidazolinidine, tetrahydrofuran-1-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydropyran-1-yl, tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, piperidin-1-yl, piperidin-4-yl, etc.

[0181] The term "aryl" refers to a monocyclic or bicyclic aromatic carbocyclic system containing 6-10 carbon atoms. Examples of aryl groups include phenyl and naphthyl groups, such as 1-naphthyl, 2-naphthyl, 3-naphthyl, and 4-naphthyl.

[0182] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic system containing a 5-10 member structure, or preferably a 5-8 member structure or a 6-10 member structure, more preferably a 5-6 member structure, wherein one, two, three, four or more ring atoms are heteroatoms and the remaining atoms are carbon atoms, the heteroatoms being independently selected from O, N or S, and the number of heteroatoms is preferably one, two, three or four. A heteroaryl group can be an aromatic monocyclic ring containing one or two 5-6 member structures selected from N, S, or O, or an aromatic fused ring containing one, two, three or four 8-10 member structures selected from N, S, or O. Examples of heteroaryl groups include, but are not limited to, furanyl, thiophene, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrroloyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, thiodiazolyl, triazinyl, phthalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purine, indoleyl, isoindoleyl, indazoleyl, benzofuranyl, benzothiophene, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, benzoimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridyl, imidazole[1, [2-a]pyridyl, pyrazolo[1,5-a]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridyl, pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, quinoline-2-yl, quinoline-3-yl, quinoline-4-yl, quinoline-5-yl, quinoline-6-yl, quinoline-7-yl, quinoline-8-yl, etc.

[0183] The term "fused ring" refers to two rings sharing two connected ring atoms. The term "linked ring" refers to two rings connected by a single bond. The term "spiral ring" refers to two rings sharing a single ring atom. The term "bridged ring" refers to two rings sharing two non-directly connected ring atoms.

[0184] In this application, the numerical ranges related to the number of substituents, carbon atoms, ring atoms, heteroatoms, and groups represent a list of all integers within that range, and the range is merely a simplified representation. For example, "1-4 substituents" means 1, 2, 3, or 4 substituents; "3-8" means 3, 4, 5, 6, 7, or 8 ring atoms. Therefore, the numerical ranges related to the number of substituents, carbon atoms, ring atoms, heteroatoms, and groups also encompass any of their subranges, and each subrange is also considered to be disclosed in this application.

[0185] The term "pharmaceutical composition" refers to a mixture containing one or more compounds described in this application and one or more pharmaceutically acceptable carriers or excipients. The pharmaceutical composition may not contain other active ingredients, or may further contain one or more other active ingredients. The purpose of the pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredients and enabling them to exert their biological activity.

[0186] The terms "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" include, but are not limited to, any flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, emulsifiers, disintegrants, binders, fillers, antioxidants, fragrances, solubilizers, osmotic pressure regulators, and the base portion of semi-solid preparations such as ointments and creams.

[0187] The pharmaceutical compositions described in this application can be prepared into liquid, solid, or semi-solid dosage forms. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can be tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, pills, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can be ointments, gels, pastes, etc. The pharmaceutical compositions of this application can be formulated into ordinary preparations, as well as sustained-release preparations, controlled-release preparations, targeted preparations, and various microparticle delivery systems.

[0188] The term "subject" refers to any human or non-human organism that may potentially benefit from treatment with the compounds described in this application. Exemplary subjects include humans or mammals of any age. Preferably, the subject is a human.

[0189] The term “treatment” includes treating a disease or symptom in mammals, particularly humans, and includes: (a) suppressing an infection, disease, or symptom, i.e., halting or delaying the development of an infection, disease, or symptom; (b) alleviating an infection, disease, or symptom, i.e. causing the remission of a disease or symptom; and / or (c) curing an infection, disease, or symptom.

[0190] The term "prevention" includes preventative therapy in mammals, particularly humans, aimed at reducing the likelihood of infection, disease, or symptoms occurring. Patients receiving preventative therapy may be selected based on an increased risk of infection or disease or symptoms compared to the general population. "Prevention" can include the management of subjects who have not yet presented with an infection or clinical condition, and the prevention of a second occurrence of the same or similar infection or clinical condition.

[0191] "Effective amount" refers to the amount of the compound of this application that, when administered alone or in combination, is effective in treating or preventing cancer, inhibiting cancer cell proliferation, and / or reducing tumor volume.

[0192] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. Those skilled in the art can make various modifications and changes to this application without departing from the scope or spirit of this application. For experimental methods in the following embodiments where specific conditions are not specified, the guidance given in this application should be used first, or experimental manuals or conventional conditions in the art can be followed, or other experimental methods known in the art can be referred to, or the conditions recommended by the manufacturer can be followed.

[0193] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0194] In addition, this application may use protecting groups to protect certain functional groups from unwanted reactions. Protecting groups suitable for various functional groups and their protection or deprotection conditions are well known to those skilled in the art. For example, TW Greene and GMWuts' "Protecting Groups in Organic Preparations" (3rd edition, Wiley, New York, 1999 and cited references in the book) describes in detail a large number of protecting or deprotecting groups.

[0195] The separation and purification of compounds and intermediates employ appropriate methods and procedures depending on specific needs, such as filtration, extraction, distillation, crystallization, column chromatography, preparative thin-layer chromatography, preparative high-performance liquid chromatography, or a combination of the above methods. Specific methods can be found in the examples described in this application. Of course, other similar separation and purification techniques can also be used. Conventional methods (including physical constants and spectroscopic data) can be used for characterization.

[0196] Preparation of NMT inhibitors

[0197] Example 1: Preparation of compound PI-7-1

[0198] Step 1: Preparation of Compound 35

[0199] Trifluoromethanesulfonic anhydride (10.0 g, 35.6 mmol, 5.88 mL, 3.50 eq) was added to a nitrogen-protected solution of dimethylacetamide (2.66 g, 30.5 mmol, 2.84 mL, 3.00 eq) in 42 mL of dichloroethane and stirred for 15 min. Then, a solution of dichloroethane (21 mL) containing compound 7 (2.10 g, 10.2 mmol, 1.00 eq) and 2,4,6-trimethylpyridine (4.75 g, 39.2 mmol, 5.18 mL, 3.85 eq) was added dropwise. After the addition was complete, the temperature was raised to 25 °C and stirred. The temperature was then raised to 90 °C and stirred for 15 h. The reaction was monitored by LC-MS. The reaction mixture was cooled to 25 °C, purified water (21 mL) was added, and the temperature was raised to 100 °C and stirred for 2 h. The reaction solution was cooled to 25°C, and the reaction was quenched with purified water (20 mL). The solution was then diluted with dichloromethane (60 mL). The organic phase was separated, washed with saturated citric acid aqueous solution (60 mL × 3), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0–4:1, TLC monitoring, Rf(P1) = 0.34, petroleum ether:ethyl acetate = 3:1) and reversed-phase HPLC (column: F-Welch Xtimate C18 40 × 200 mm, 7 μm; mobile phase A: H2O (0.225% FA), B: ACN, within 20.0 min, B: 28%–68%) to give compound 35 (340 mg, 1.37 mmol, 13.5% yield) as a white solid.

[0200] LCMS(ESI): m / z, 249.3 [M+H] + Rt = 0.727 min.

[0201] 1 H NMR: (400MHz, CDCl3) δppm 3.83 (s, 3H), 3.71 (tt, J = 9.40, 7.60Hz, 1H), 3.23-3.44 (m, 4H), 2.28 (s, 3H), 1.39 (s, 9H).

[0202] Step 2: Preparation of Compound 36

[0203] At 0 °C, a solution of sodium borohydride (100 mg, 2.64 mmol, 2.74 eq) and CeCl3 (262 mg, 1.06 mmol, 66.9 μL, 1.10 eq) in methanol (5 mL) was added to a tetrahydrofuran (5 mL) solution of compound 35 (240 mg, 966 μmol, 1.00 eq). After the addition was complete, the reaction mixture was reacted at 25 °C for 2 hours, and LC-MS showed that the reaction was essentially complete. The mixture was cooled to 0 °C, diluted with an aqueous solution of ammonium chloride, and the aqueous phase was extracted with ethyl acetate (3 × 40 mL). The combined organic phases were dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under vacuum to give the crude product. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0-1:2, TLC monitoring, Rf(P1) = 0.5, petroleum ether: ethyl acetate = 0:1) to give compound 36 (205 mg, 812 μmol, 84.0% yield) as a white solid.

[0204] LCMS(ESI): m / z, 253.2 [M+H] + , Rt=0.400min.

[0205] 1 H NMR: (400MHz, CDCl3) δppm 4.35 (s, 1H), 4.23 (q, J = 7.40Hz, 1H), 3.72 (s, 3H), 2.79-2.90 (m, 1H), 2.65-2.78 (m, 2H), 2.54 (br s,1H),2.03-2.17(m,2H),1.84-2.01(m,1H).

[0206] Step 3: Preparation of Compound 37

[0207] Triphenylphosphine (455 mg, 1.74 mmol, 1.10 eq) and DIAD (diisopropyl azodicarbonate, 351 mg, 1.74 mmol, 337 μL, 2.60 eq) were added to a tetrahydrofuran (5 mL) solution of compounds 36 (185 mg, 734 μmol, 1.10 eq) and 16 (260 mg, 668 μmol, 1.00 eq) at 0 °C. The mixture was heated to 35 °C and stirred for 12 hours. LC-MS showed that the starting materials reacted completely. The reaction was quenched with 10 mL of purified water. The reaction solution was diluted with ethyl acetate (20 mL). The organic phase was washed with purified water (10 mL × 2) and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under vacuum to give the crude product. The crude product was purified twice by reversed-phase HPLC (column: F-Welch Xtimate C18 40×200mm, 7μm; mobile phase A: H2O (0.225% FA), B: ACN, within 20.0 min, B: 12%-52%; column: YMC Triart C18 30×250mm, 5μm; mobile phase A: H2O (0.225% FA), B: MeOH, within 10.0 min, B: 50%-80%) to obtain compound 37 (55.0 mg, 88.2 μmol, 13.2% yield) as a white solid.

[0208] LCMS(ESI): m / z, 624.5 [M+H] + Rt = 0.682 min.

[0209] 1 H NMR: (400MHz, DMSO-d6) δppm 8.57(s,1H),7.61-7.72(m,2H),7.49(br d,J=9.20Hz,1H),7.30(s,2H),4.72-4.92(m,3H),4.35(d,J=5.20Hz,1H),3.96(d,J=5.20Hz,1 H),3.57(s,4H),2.63-2.70(m,3H),2.18-2.37(m,4H),2.07(s,3H),1.36(s,9H),0.66(s,9H).

[0210] 19 F NMR: (400MHz, DMSO-d6) δppm-136.66,-153.83,-153.89.

[0211] Step 4: Preparation of compound PI-7-1

[0212] Compound 37 (45.0 mg, 72.2 μmol, 1.00 eq) was reacted with a nitrogen-protected solution of HCl / EtOAc (2.00 M, 1.50 mL, 41.6 eq) at 25 °C for 8 h. LC-MS showed the reaction was complete. The mixture was filtered. The filter cake was washed with ethyl acetate (10 mL) and dried to give compound PI-7-1 (22.4 mg, 39.99 μmol, 55.44% yield) as a white solid.

[0213] LCMS(ESI): m / z, 524.3 [M+H] + Rt = 0.472 min.

[0214] HPLC: Rt=2.548min, 99.2%.

[0215] 1 H NMR(MeOH-d4)δppm 9.27(s,1H),8.42(s,2H),8.10(d,J=7.80Hz,1H),7.60(s,1H),7.23(d,J=6.80Hz,1H),4. 95-5.20(m,3H),4.43(s,1H),3.69-3.98(m,4H),2.88(s,3H),2.21-2.66(m,7H),0.84(br s,8H).

[0216] Example 2: Preparation of compound PI-7-2

[0217] Step 1: Preparation of Compound 38

[0218] Compound 6 (200 mg, 772 μmol, 1.00 eq), 3-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)cyclopent-2-en-1-one (177 mg, 849 μmol, 1.10 eq), Pd(dtbpf)Cl2 (1,1'-bis(di-tert-butylphosphino)ferrocene dichloropalladium, 39.4 mg, 77.2 μmol, 0.100 eq), and potassium carbonate (213 mg, 1.54 mmol, 2.00 eq) in dioxane (5 mL) and purified water (0.8 mL) were degassed and purged with nitrogen three times. The mixture was heated to 80 °C and stirred for 14 hours; LC-MS showed the reaction was complete. The mixture was cooled to 25 °C, and purified water (15 mL) was added to quench the reaction. The mixture was then diluted with ethyl acetate (20 mL). The aqueous phase was extracted with ethyl acetate (25 mL × 2). The organic phases were combined and washed with brine (40 mL), then dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under vacuum to give the crude product. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0–1:1, TLC monitoring, Rf(P1) = 0.49, petroleum ether:ethyl acetate = 1:1) to give compound 38 (0.399 g, crude product) as a white solid.

[0219] LCMS(ESI): m / z, 261.3 [(M+H)] + RT = 0.660 min.

[0220] 1 H NMR (CDCl3) δppm 6.02 (t, J = 2.00Hz, 1H), 3.87 (s, 3H), 2.86 (td, J = 4.80, 2.00Hz, 2H), 2.43-2.58 (m, 2H), 2.27 (s, 3H), 1.38 (s, 9H).

[0221] Step 2: Preparation of compound 39

[0222] Pd(OH)₂ / C (108 mg, 154 μmol, 20% purity, 0.200 eq) and hydrogen gas (45 psi) were added to a tetrahydrofuran (5 mL) solution of compound 38 (200 mg, 768 μmol, 1.00 eq), and the mixture was heated to 45 °C and stirred for 16 hours. LC-MS showed that the starting material reacted completely. The reaction solution was filtered, and the filtrate was concentrated under vacuum to give compound 39 (190 mg, 724 μmol, yield 94.3%) as a colorless oil.

[0223] LCMS(ESI): m / z, 263.1 [(M+H)] + RT = 0.660 min.

[0224] 1 H NMR(CDCl3)δppm 3.83(s,3H),3.51-3.63(m,1H),2.65-2.76(m,1H),2.43-2.53(m,1H), 2.32-2.40(m,1H),2.19-2.30(m,4H),2.07-2.15(m,1H),1.39(s,9H).

[0225] Step 3: Preparation of Compound 40

[0226] Sodium borohydride (80.0 mg, 2.11 mmol, 2.93 eq) was added to a tetrahydrofuran (5 mL) solution of compound 39 (189 mg, 722 μmol, 1.00 eq) at 25 °C, and the mixture was stirred for 16 hours. The mixture was then cooled to 0 °C, and a methanol (5 mL) solution of CeCl3 (196 mg, 794 μmol, 1.10 eq) was added to the reaction mixture. After the addition was complete, the reaction mixture was allowed to react at 25 °C for 2 hours. The mixture was then cooled to 0 °C, diluted with an aqueous solution of ammonium chloride (10 mL), and the aqueous phase was extracted with ethyl acetate (3 × 40 mL). The combined organic phases were dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under vacuum to give the crude product. The crude product was purified by silica gel column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0-1:2, TLC monitoring, Rf(P1) = 0.5, petroleum ether: ethyl acetate = 0:1) to give compound 40 (138 mg, 518 μmol, yield 71.7%), which was a white gel.

[0227] LCMS(ESI): m / z, 267.1 [M+H] + Rt = 0.359 min.

[0228] 1 H NMR: (400MHz, CDCl3) δppm 4.32-4.55(m,2H),3.72(s,3H),2.84-3.45(m,1H),2.07-2.36(m,3H),1.57-1.93(m,6H),0.91-0.99(m,9H).

[0229] Step 4: Preparation of Compound 41

[0230] Triphenylphosphine (175 mg, 668 μmol, 2.60 eq) and DIAD (135 mg, 668 μmol, 129 μL, 2.60 eq) were added to a tetrahydrofuran (3 mL) solution of compounds 40 (75.3 mg, 282 μmol, 1.10 eq) and 16 (100 mg, 257 μmol, 1.00 eq) at 0 °C. The mixture was heated to 35 °C and stirred for 12 hours. LC-MS showed that the starting material reacted completely. The reaction was quenched with 10 mL of purified water. The reaction solution was diluted with ethyl acetate (10 mL), and the aqueous phase was extracted with ethyl acetate (25 mL × 2). The combined organic phases were washed with brine (25 mL) and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under vacuum to give the crude product. The crude product was purified by reversed-phase HPLC (column: Prepulite XP tC18 40×200mm, 7μm; mobile phase A: H2O (10mM NH4HCO3), B: ACN, within 20.0 min, B: 50%-90%) to give compound 41 (139mg, 218μmol, yield 84.8%) as a white solid.

[0231] LCMS(ESI): m / z, 638.3 [M+H] + Rt = 0.599 min.

[0232] 1 H NMR: (400MHz, DMSO-d6) δppm 8.26-8.70(m,1H),7.58-7.72(m,2H),7.39-7.51(m,1H),7.27(s,2H),4.53-4.87(m,3H),4.30-4.49(m,1H),4.0 3-4.17(m,1H),3.50-3.61(m,3H),3.00-3.26(m,1H),2.66(s,3H),1.44-1.98(m,9H),1.29-1.39(m,9H),0.78(br d,J=5.60Hz,9H).

[0233] 19 F NMR: (400MHz, DMSO-d6) δppm-136.7,-153.3.

[0234] Step 5: Preparation of compound PI-7-2

[0235] Compound 41 (89.0 mg, 140 μmol, 1.00 eq) was reacted with a nitrogen-protected solution of HCl / EtOAc (2.00 M, 2.50 mL, 35.8 eq) at 25 °C for 6 h. LC-MS showed the reaction was complete. The mixture was filtered. The filter cake was washed with ethyl acetate (10 mL) and lyophilized to give compound PI-7-2 (59.9 mg, 104 μmol, yield 74.8%) as a yellow solid.

[0236] LCMS(ESI): m / z, 538.1 [M+H] + Rt = 0.429 min.

[0237] HPLC: Rt = 2.711 min, purity 99.2%.

[0238] 1 H NMR(DMSO-d6)δppm 9.92(br s,2H),9.31(d,J=2.80Hz,1H),8.42(s,1H),8.16-8.26(m,1H),8.07(d,J=9.40Hz,1H),7.61-7.76(m,1H),7.26-7.52(m,1H),4.70-4.89(m ,3H),4.21-4.37(m,2H),3.71-3.75(m,3H),3.28-3.36(m,1H),2.59- 2.68(m,3H),2.09-2.27(m,3H),1.45-2.06(m,6H),0.71-0.85(m,9H).

[0239] Example 3: Preparation of compounds PI-7-1iP-P1 and P2

[0240] Step 1: Synthesis of compound 7-1iP-b

[0241] At 0 °C, trifluoromethanesulfonic anhydride (4.84 g, 17.164 mmol) was added to a solution of N,N-dimethylacetamide (1.36 g, 15.604 mmol) in 60 mL of 1,2-dichloroethane. The mixture was then stirred at 0 °C for 10 min to obtain a reaction solution. At 15 °C, 2,4,6-trimethylpyridine (2.52 g, 20.805 mmol) and 1-(1,5-dimethyl-4-vinylpyrazole-3-yl)-2-methylprop-1-one (1 g, 5.201 mmol) were dissolved in 10 mL of 1,2-dichloroethane to obtain a mixed solution. The mixed solution was added dropwise to the reaction solution at 0 °C. The mixture was then stirred at 90 °C for 18 h. LC-MS showed that the reaction was complete. The reaction solution was diluted with saturated sodium bicarbonate aqueous solution (50 mL), and EtOAc (100 mL × 2) was added for extraction. The separated organic phase was concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography with silica gel (40 g, PE:EtOAc = 37%:63%) to obtain 7-1iP-b (570 mg, 2.433 mmol, yield 46.8%), which was a yellow oil.

[0242] LCMS (ESI): m / z, 234.9 [M] + Rt = 1.297 min.

[0243] Step 2: Preparation of compound 7-1iP-c

[0244] At 0 °C, cerium trichloride (55 mg, 0.222 mmol) and sodium borohydride (84 mg, 2.219 mmol) were added to an ethanol (10 mL) solution of 7-1iP-b (520 mg, 2.219 mmol). The mixture was then stirred at 40 °C for 30 minutes. LC-MS showed that the reaction was complete. The reaction solution was diluted with water (20 mL), extracted with ethyl acetate (50 mL × 2), and the aqueous phase was concentrated under vacuum to give the crude product. The crude product was purified by column chromatography with silica gel (40 g, PE:EtOAc = 27%:73%) to give 7-1iP-c (311 mg, 0.921 mmol, yield 41.5%) as a colorless oil.

[0245] LCMS (ESI): m / z, 236.7 [M] + , Rt=1.201min.

[0246] Step 3: Preparation of compound 7-1iP-d

[0247] Compound 16 (358 mg, 0.918 mmol), triphenylphosphine (482 mg, 1.837 mmol), and ethyl [(1E)-(ethoxycarbonyl)diazeline]formate (320 mg, 1.83 mmol) were added to a solution of 7-1iP-c (45 mg, 0.113 mmol) in 10 mL of dioxane at 15 °C. The mixture was then stirred at 100 °C for 16 hours under nitrogen. LC-MS showed that the reaction was complete. The reaction solution was diluted with water (20 mL), extracted with ethyl acetate (50 mL × 2), and the aqueous phase was concentrated under vacuum to give the crude product. The crude product was purified by column chromatography on C18 (0.1% TFA, H2O:ACN = 53%:47%) to give 7-1iP-d (311 mg, 0.358 mmol, yield 39%) as a colorless oily liquid.

[0248] LCMS (ESI): m / z, 608.0 [M+H] + , Rt=2.559min.

[0249] Step 4: Preparation of compound 7-1iP-e

[0250] At 0 °C, cerium trichloride (18 mg, 0.072 mmol) and sodium borohydride (14 mg, 0.358 mmol) were added to a methanol (6 mL) solution of 7-1iP-d (311 mg, 0.358 mmol). The mixture was then stirred at 15 °C for 2 hours. LC-MS showed that the reaction was complete. The reaction solution was diluted with water (10 mL), extracted with ethyl acetate (50 mL × 2), and the separated organic phase was concentrated under vacuum to obtain the crude product.

[0251] The crude product was purified by silica gel column chromatography (10 g, DCM:MeOH = 6%:94%) to give a yellow oily 7-1iP-e (100 mg, 0.164 mmol, yield 46%).

[0252] LCMS (ESI): m / z, 610.0 [M] + Rt = 1.342 min.

[0253] Step 5: Preparation of compounds PI-7-1iP-P1 and PI-7-1iP1-P2

[0254] At 15 °C, HCl-dioxane (2 mL, 4 M) was added to a solution of 7-1iP-e (100 mg, 0.164 mmol) in dioxane (2 mL). The mixture was then stirred at 15 °C for 30 min. LC-MS showed that the reaction was complete. The reactants were purified by high performance liquid chromatography (column: JB-C235-04, Prime C18, 30 × 150 mm, 5 μm; mobile phase A: 0.1% TFA, H2O; phase B: ACN, within 14 min, B: 15%–45%) to obtain the crude product. The crude product was then purified using Waters PrepSFC Daicel ChiralPak IC, 40 mm × 250 mm, 10 μm (A phase: supercritical CO2, B phase: 0.1% NH3·H2O (70% EtOH), A:B = 35%:65%) to obtain white solid PI-7-1iP-P1 (10.25 mg, 0.020 mmol, yield 12.26%) and PI-7-1iP-P2 (11.47 mg, 0.023 mmol, yield 13.72%).

[0255] LCMS (ESI): m / z, 510.7 [M+H] + Rt = 1.276 min.

[0256] 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.92(s,2H),8.11(s,1H),7.97(q,J=9.3Hz,2 H),7.50–7.35(m,2H),4.92(s,2H),4.72(s,3H),3.94(d,J=8.1Hz,1H),3.67–3.54( m, 4H), 2.66 (t, J = 4.8 Hz, 3H), 2.48–2.36 (m, 2H), 2.30 (s, 1H), 2.11 (s, 3H), 1.81 (dd, J = 14.5, 6.8 Hz, 1H), 0.88 (t, J = 9.0 Hz, 3H), 0.59 (dd, J = 14.7, 6.8 Hz, 3H). (racemic data).

[0257] Retention time on SFC of PI-7-1iP-P1: 4.499 min (IC25% EtOHB), Retention time on SFC of PI-7-1iP-P2: 6.059 min (IC25% EtOHB).

[0258] Example 4: Preparation of compound PI-H7-3iP-1,-2

[0259] Step 1: Preparation of compound H7-3iP-b

[0260] At room temperature, acetic acid (0.621 mL, 11.035 mmol, 5.0 eq) was added to an ethanol (30 mL) solution of compound H7-3iP-S (400 mg, 2.048 mmol, 1.0 eq) and compound 3-oxoacetic acid cyclobutyl ester (990 mg, 7.725 mmol, 2.0 eq), and the mixture was stirred at room temperature for 20 minutes. After 20 minutes, NaBH3CN (555 mg, 8.828 mmol, 4.0 eq) was added to the reaction system, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by C18 reversed-phase column chromatography (column: JB-C235-04, Prime C18, 30×150mm, 5μm; mobile phase: A: 0.1% TFA, H2O; B: ACN, within 30 min, B: 5%-80%) to obtain a yellow solid H7-3iP-b (240mg, yield 26%, purity 70%).

[0261] LCMS(ESI): m / z, 294.2 [M+H] + Rt = 3.77 min.

[0262] Step 2: Preparation of compound H7-3iP-c

[0263] At room temperature, LiOH hydrate (41.19 mg, 0.982 mmol, 1.71 eq) was added to a methanol (10.0 mL) solution of compound H7-3iP-b (240 mg, 70% purity, 0.573 mmol, 1.0 eq). The reaction solution was stirred at 70 °C for 2 hours under nitrogen atmosphere, and then allowed to return to room temperature. Boc₂O (886 mg, 4.06 mmol, 7.1 eq) was then added, and the reaction solution was stirred overnight at room temperature. The reaction solution was purified by C18 reversed-phase column chromatography (column: JB-C235-04, Prime C18, 30 × 150 mm, 5 μm; mobile phase: A: 0.1% TFA, H₂O; B: ACN, within 25 min, B: 5%-70%) to obtain a yellow solid H7-3iP-c (92 mg, two-step yield 23%).

[0264] LCMS(ESI): m / z, 374.2 [M+Na] + Rt = 1.56 min.

[0265] Step 3: Preparation of compounds H7-3iP-d1,-d2

[0266] Compound H7-3iP-c (50.0 mg, 0.142 mmol, 1.0 eq), compound 16 (55.4 mg, 0.142 mmol, 1.0 eq), and triphenylphosphine (112 mg, 0.426 mmol, 3.0 eq) were dissolved in 1,4-dioxane (5.0 mL), followed by the addition of diethyl azodicarbonate (74 mg, 0.426 mmol, 3.0 eq), and the mixture was purged under nitrogen atmosphere. The reaction mixture was stirred overnight at 100 °C. The reaction solution was filtered and then directly purified by high-performance liquid chromatography (HPLC) (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm; mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 20%-70%) to obtain two isomers. The first eluting isomer was designated H7-3iP-d1 (7.95 mg, yield 19.3%, yellow solid); the second eluting isomer was designated H7-3iP-d2 (15.18 mg, yield 36.8%, yellow solid). These should be cis-trans isomers.

[0267] LCMS(ESI):H7-3iP-d1 m / z, 723.4[M+H] + .

[0268] LCMS(ESI):H7-3iP-d2 m / z, 723.4[M+H] + .

[0269] Step 4: Preparation of compound H7-3iP-e1

[0270] Compound H7-3iP-d1 (7.95 mg, 0.011 mmol, 1.0 eq) was dissolved in methanol (2.0 mL), sodium borohydride (4.2 mg, 0.11 mmol, 10.0 eq) and cerium trichloride (0.5 mg, 0.0022 mmol, 0.2 eq) were added, and the mixture was purged under nitrogen. The reaction mixture was stirred overnight at room temperature. The reaction mixture was quenched with water (5.0 mL) and extracted with ethyl acetate (5.0 mL × 3). The solution was dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure and used directly in the next step.

[0271] LCMS (ESI): m / z, 725.4 [M+H] + .

[0272] Step 5: Preparation of compound PI-H7-3iP-1

[0273] At room temperature, the crude product of compound H7-3iP-e1 was dissolved in 2 mL of 4 M HCl in a 1,4-dioxane solution and reacted at room temperature for one hour. The reaction solution was concentrated under reduced pressure, and the crude product was purified by high performance liquid chromatography (HPLC) (column: JB-C235-04, Bonnasil-BS C18, 20×250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 20 min, B: 20%-80%) to obtain a yellow solid product PI-H7-3iP-1 (6.05 mg, two-step yield 86.12%).

[0274] LCMS(ESI): m / z, 525.4 [(M+H)] + .

[0275] 1 H NMR (400MHz, DMSO-d) 6) δ8.89(s,3H),7.98(s,1H),7.82(d,J=9.4Hz,1H),7.69(d,J=9.4Hz,1H),7.40(t,J=6.2Hz,2H),4.68(s,2H),4.32–4.26(m,2H),3 .64(s,3H),2.64(s,3H),2.21(s,1H),2.16(s,1H),2.09–1.93(m,3H),1.23(s,2H),0.88(d,J=6.8Hz,3H),0.76(d,J=6.8Hz,3H).

[0276] Step 4-2: Preparation of compound H7-3iP-e2

[0277] Compound H7-3iP-d2 (15 mg, 0.021 mmol, 1.0 eq) was dissolved in methanol (2 mL), sodium borohydride (8 mg, 0.210 mmol, 10.0 eq) and cerium trichloride (1 mg, 0.0042 mmol, 0.2 eq) were added, and the mixture was purged under nitrogen. The reaction mixture was stirred overnight at room temperature. LC-MS showed that the starting material reacted completely. The reaction mixture was quenched with water (5 mL), extracted with ethyl acetate (5 mL × 3), dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure and used directly in the next step.

[0278] LCMS(ESI): m / z, 725.4 [M+H] + .

[0279] Step 5-2: Preparation of compound PI-H7-3iIP-2

[0280] At room temperature, the crude product of H7-3iP-e2 was dissolved in 1,4-dioxane solution (2 mL) of 4M HCl and reacted at room temperature for one hour. LCMS showed that the starting material reacted completely. The reaction solution was concentrated under reduced pressure, and the crude product was purified by high performance liquid chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 14 min, B: 20%-80%) to obtain a yellow solid product PI-H7-3iP-2 (9.95 mg, two-step yield 74.19%).

[0281] LCMS(ESI): m / z, 525.4 [(M+H)] + .

[0282] 1 H NMR (400MHz, DMSO-d6) δ8.92(s,3H),8.00(s,1H),7.83(d,J=9.2Hz,1H),7 .75(d,J=9.2Hz,1H),7.44–7.37(m,2H),4.87(s,1H),4.68(s,2H),4.27(d, J=6.5Hz,2H),3.64(s,3H),2.64(s,3H),2.36–2.19(m,3H),2.14(s,3H),2. 10–1.92(m,3H),1.23(s,2H),0.87(d,J=6.8Hz,3H),0.75(d,J=6.8Hz,3H).

[0283] Example 5: Preparation of compound PI-10-6MeiP

[0284] Step 1: Preparation of compound 10-6MeiP-b

[0285] At room temperature, acetic acid (1.087 mL, 19.311 mmol, 5.0 eq) was added to a 20.0 mL ethanol solution of compound H7-3iP-S (700 mg, 3.862 mmol, 1.0 eq) and tert-butyldimethylsiloxane acetaldehyde (673.23 mg, 3.862 mmol, 1.0 eq), and the mixture was stirred at room temperature for 20 minutes. Subsequently, sodium borohydride (2427.06 mg, 38.623 mmol, 10.0 eq) was added to the reaction mixture, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 30 min, B: 5%-80%) to obtain a yellow oily substance 10⁻⁶MeiP-b (342mg, yield 26%).

[0286] LCMS(ESI): m / z, 240.2 [M+H] + Rt = 1.445 min.

[0287] Step 2: Preparation of compound 10-6MeiP-c

[0288] At room temperature, acetic acid (0.282 mL, 5.01 mmol, 5.0 eq) was added to a 6.0 mL ethanol solution of compound 10⁻⁶MeiP-b (170 mg, 0.501 mmol, 1.0 eq) and an aqueous solution of formaldehyde (138 μL, 5.01 mmol, 10.0 eq, 35–40%, 1.090 g / mL), and the mixture was stirred at room temperature for 20 minutes. Subsequently, sodium borohydride (157.30 mg, 2.503 mmol, 5.0 eq) was added to the reaction mixture, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 5%-70%) to obtain a yellow oily substance 10⁻⁶MeiP-c (103mg, yield 58.19%).

[0289] LCMS (ESI): m / z, 354.4 [M+H] + Rt = 5.095 min.

[0290] Step 3: Preparation of compound 10-6MeiP-d

[0291] At room temperature, a solution of 10⁻⁶MeiP-c (93 mg, 0.263 mmol, 1.0 eq) in tetrahydrofuran (2.0 mL) was added to a solution of hydrofluoric acid / pyridine (260.68 mg, 2.630 mmol, 10.0 eq), and the mixture was stirred at room temperature for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was directly purified by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20 × 250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 5%–80%) to give a yellow oily substance 10⁻⁶MeiP-d (23.4 mg, 37.17% yield).

[0292] LCMS (ESI): m / z, 240.5 [M+H] + Rt = 1.176 min.

[0293] Step 4: Preparation of compound 10-6MeiP-e

[0294] Compound 10-6MeiP-d (23.4 mg, 0.098 mmol, 1.0 eq), compound 16 (38.07 mg, 0.098 mmol, 1.0 eq), and triphenylphosphine (76.94 mg, 0.293 mmol, 3.0 eq) were dissolved in 1,4-dioxane (3.0 mL), followed by the addition of diethyl azodicarbonate (51.09 mg, 0.293 mmol, 3.0 eq), and the mixture was purged under nitrogen atmosphere. The reaction mixture was stirred overnight at 100 °C. LC-MS showed that the starting materials reacted completely. The reaction solution was filtered and then purified directly by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 5%-70%) to obtain a yellow solid 10-6MeiP-e (28mg, 46.89% yield).

[0295] LCMS (ESI): m / z, 611.6 [M+H] + Rt = 1.369 min.

[0296] Step 5: Preparation of compound 10-6MeiP-f

[0297] At room temperature, compound 10⁻⁶MeiP-e (28 mg, 0.046 mmol, 1.0 eq) was dissolved in methanol (3.0 mL), and sodium borohydride (17.34 mg, 0.460 mmol, 10.0 eq) was added. The reaction mixture was reacted at 50 °C for two hours. The reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was used directly in the next step.

[0298] LCMS (ESI): m / z, 613.2 [M+H] + Rt = 1.30 min.

[0299] Step 6: Preparation of compound PI-10-6MeiP

[0300] The crude product from the previous step was dissolved in 4M HCl in a 1,4-dioxane solution (4 mL) and reacted at room temperature for one hour. LCMS showed that the starting material reacted completely, and the reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was purified by high performance liquid chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 14 min, B: 20%-80%) to obtain a white solid product PI-10-6MeiP (4.08 mg, two-step yield 17.41%).

[0301] LCMS(ESI): m / z, 513.4, [M+H] + Rt = 1.332 min.

[0302] 1 H NMR (400MHz, DMSO-d6) δ8.86(s,1H),8.75(s,2H),7.96(s,1H),7.78(s,2H),7.42–7.35(m,2H),5.42–5.23(m,1H),4.66(s,2H),4.05–3.99(m ,3H),3.12(t,J=5.9Hz,2H),2.63(t,J=5.1Hz,3H),2.58(s,3H),2.03(s,3H),2.01–1.94(m,4H),0.86(d,J=6.7Hz,3H),0.62(d,J=6.7Hz,3H).

[0303] Example 6: Preparation of compound PI-10-6iP

[0304] Step 1: Preparation of compound 10-6iP-c

[0305] At room temperature, LiOH hydrate (21.01 mg, 0.501 mmol, 1.0 eq) and di-tert-butyl dicarbonate (1092.66 mg, 5.006 mmol, 10.0 eq) were added to a methanol (7.0 mL) / water (3.0 mL) mixture of compound 10-6MeiP-b (170 mg, 0.501 mmol, 1.0 eq). The reaction mixture was stirred at room temperature for 18 hours. LC-MS showed that the starting material reacted completely. The reaction mixture was purified by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20 × 250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 30 min, B: 5%–70%) to give a yellow solid 10-6iP-c (115.0 mg, yield 52.24%).

[0306] LCMS(ESI): m / z, 462.4 [M+Na] + Rt = 2.22 min.

[0307] Step 2: Preparation of compound 10-6iP-d

[0308] A solution of compound 10-6iP-c (105 mg, 0.239 mmol, 1.0 eq) and TABF (tetrabutylammonium fluoride, 2.0 mL, 1 M THF solution, 8.4 eq) was stirred at room temperature for two hours. The reaction solution was quenched with water (10 mL) and extracted with ethyl acetate (10 mL × 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel normal-phase column chromatography (CH3OH:CH2Cl2 = 0:100-5:95) to give a yellow oily substance 10-6iP-d (73 mg, yield 93.94%).

[0309] LCMS(ESI): m / z, 226.2[M-Boc+H] + Rt = 1.30 min.

[0310] Step 3: Preparation of compound 10-6iP-e

[0311] Compound 10-6iP-d (73 mg, 0.224 mmol, 1.0 eq), compound 16 (95.95 mg, 0.246 mmol, 1.1 eq), and triphenylphosphine (176.26 mg, 0.672 mmol, 3.0 eq) were dissolved in 1,4-dioxane (10 mL), followed by the addition of diethyl azodicarbonate (117.04 mg, 0.672 mmol, 3.0 eq), and the mixture was purged under nitrogen atmosphere. The reaction mixture was stirred overnight at 100 °C. LC-MS showed that the starting materials reacted completely. The reaction solution was filtered and then purified directly by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 5%-60%) to obtain a yellow solid 10-6iP-e (150mg, yield 57.58%).

[0312] LCMS (ESI): m / z, 697.5 [M+H] + Rt = 1.41 min.

[0313] Step 4: Preparation of compound 10-6iP-f

[0314] At room temperature, compound 10-6iP-e (150 mg, 0.215 mmol, 1.0 eq) was dissolved in methanol (4.0 mL), and sodium borohydride (81.33 mg, 2.150 mmol, 10.0 eq) was added. The reaction mixture was reacted at 50 °C for two hours. The reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (10 mL × 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was used directly in the next step.

[0315] LCMS (ESI): m / z, 699.4 [M+H] + Rt = 1.269 min.

[0316] Step 5: Preparation of compound PI-10-6iP

[0317] The crude product from the previous step was dissolved in 4M HCl in a 1,4-dioxane solution (4 mL) and reacted at room temperature for one hour. LCMS showed that the starting material reacted completely, and the reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was purified by high performance liquid chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 14 min, B: 25%-75%) to obtain a white solid product PI-10-6iP (31.94 mg, two-step yield 29.84%).

[0318] PI-10-6iP: LCMS(ESI): m / z, 499.4[(M+H)] + Rt = 0.808 min.

[0319] 1 H NMR (400MHz, DMSO) δ8.98(s,1H),8.07(s,1H),7.92–7.81(m,2H),7.48–7.38(m,2H),4.70(s,2H),4.28–4.20(m,3H), 3.62(s,3H),3.27(s,2H),2.64(s,3H),2.11(s,3H),2.01–1.93(m,1H),0.84(d,J=6.7Hz,3H),0.73(d,J=6.8Hz,3H).

[0320] Example 7: Preparation of compounds PI-H7-3iPCH2 and PI-H7-3iPCH2dH

[0321] Step 1: Preparation of compound H7-3iPCH2-b

[0322] At room temperature, acetic acid (745 mg, 12.414 mmol, 5.0 eq) was added to ethanol (5.0 mL) containing compound H7-3iP-S (450 mg, 2.483 mmol, 1.0 eq) and 3-(hydroxymethyl)cyclobutane-1-one (870.06 mg, 8.690 mmol, 3.5 eq), and the mixture was stirred at room temperature for 20 minutes. After 20 minutes, NaBH3CN (1560 mg, 24.829 mmol, 10.0 eq) was added to the reaction mixture, and the mixture was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 30 min, B: 5%-80%) to obtain a yellow solid H7-3iPCH2-b (600mg, 36% yield, 40% purity).

[0323] LCMS(ESI): m / z, 266.2 [M+H] + Rt = 1.93 min.

[0324] Step 2: Preparation of compound H7-3iPCH2-c

[0325] At room temperature, LiOH hydrate (38 mg, 0.904 mmol, 1.0 eq) and di-tert-butyl dicarbonate (1973 mg, 9.040 mmol, 10.0 eq) were added to a methanol (7.0 mL) / water (3.0 mL) solution of compound H7-3iPCH2-b (600 mg, 40% purity, 0.904 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for 18 hours. LC-MS showed that the starting material reacted completely. The reaction mixture was purified by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20 × 250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 5%–70%) to give a yellow solid H7-3iPCH2-c (90 mg, yield 27%).

[0326] LCMS(ESI): m / z, 266.2[M-Boc+H] + Rt = 1.14 min.

[0327] Step 3: Preparation of compound H7-3iPCH2-d

[0328] Compound H7-3iPCH2-c (45 mg, 0.123 mmol, 1.0 eq), compound 16 (47.95 mg, 0.123 mmol, 1.0 eq), and triphenylphosphine (97.0 mg, 0.369 mmol, 3.0 eq) were dissolved in 1,4-dioxane (3.0 mL), followed by the addition of diethyl azodicarbonate (64.3 mg, 0.369 mmol, 3.0 eq), and the mixture was purged under nitrogen atmosphere. The reaction mixture was stirred overnight at 100 °C. LC-MS showed that the starting materials reacted completely. The reaction solution was filtered and then purified directly by C18 reversed-phase column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 25 min, B: 5%-70%) to obtain a yellow solid H7-3iPCH2-d (67mg, 59.08% yield).

[0329] LCMS(ESI): m / z, 737.4 [M+H] + Rt = 1.49 min.

[0330] Step 4: Preparation of compound PI-H7-3iPCH2dH

[0331] Compound H7-3iPCH2-d (67 mg, 0.073 mmol, 1.0 eq) was dissolved in 4 M HCl in a 1,4-dioxane solution (4 mL) and reacted at room temperature for one hour. LC-MS showed that the starting material reacted completely. The reaction solution was concentrated under reduced pressure, and the crude product was used directly in the next step.

[0332] LCMS(ESI): m / z, 537.4 [M+H] + Rt = 1.48 min.

[0333] Step 5: Preparation of compound PI-H7-3iPCH2

[0334] At room temperature, the crude PI-H7-3iPCH2dH was dissolved in methanol (2.0 mL), and sodium borohydride (9 mg, 0.219 mmol, 3.0 eq) and cerium trichloride (3.6 mg, 0.015 mmol, 0.2 eq) were added. The reaction solution was reacted at 40 °C for one hour. LCMS showed that half of the starting material was consumed. The reaction solution was concentrated under reduced pressure, and the crude product was purified by high performance liquid chromatography (column: JB-C235-04, Bonnasil-BS C18, 20 × 250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 14 min, B: 20%-70%) to obtain a white solid product PI-H7-3iPCH2 (6.45 mg, two-step yield 16.40%). The white solid PI-H7-3iPCH2dH was recovered (4.99 mg, recovery rate 12.74%).

[0335] LCMS(ESI): m / z, 539.3 [M+H] + Rt = 0.77 min.

[0336] 1 H NMR (400MHz, DMSO) δ9.11–8.82(m,3H),7.97(d,J=6.0Hz,1H),7.80(t,J=13.8Hz,1H),7.72(s,1H),7.46–7.37(m,2H),4.67(s,2H),4.36(t,J=6.8H z,1H),4.02–3.92(m,3H),3.69(s,2H),3.67(s,1H),2.63(s,3H),2.34–2 .19(m,5H),2.12–1.87(m,4H),1.23(s,1H),0.85(dd,J=41.2,6.8Hz,6H).

[0337] Example 8: Preparation of compound PI-R1dH

[0338] Step 1: Preparation of N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide

[0339] 1,5-Dimethyl-1H-pyrazole-3-carboxylic acid (105 g, 749.2 mmol) was dissolved in dichloromethane (1000 mL), and oxalyl chloride (66 mL, 786.7 mmol) and triethylamine (219 mL, 1573.4 mmol) were added at room temperature. The reaction mixture was stirred at 25 °C for 1 hour. Then, dimethylhydroxylamine hydrochloride (73.1 g, 749.2 mmol) was added in portions. Stirring was continued for 16 hours, and the reaction solution was slowly poured into water and stirred until room temperature. Then, dichloromethane (300 mL × 3) was added for extraction. The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (EtOAc:PE = 0:100-10:90) to give a brown solid compound N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide (112 g, yield 81.6%).

[0340] LCMS (ESI): m / z, 184.2 [M+H] + Rt = 1.20 min.

[0341] 1 H NMR (400MHz, DMSO-d6) δ6.41 (s, 1H), 3.77 (d, J = 2.3Hz, 3H), 3.67 (s, 3H), 3.30 (s, 3H), 2.26 (s, 3H).

[0342] Step 2: Preparation of compound 1-(1,5-dimethyl-1H-pyrazole-3-yl)-2,2-dimethylprop-1-one

[0343] At -70°C, tert-butyllithium (141 mL, 1.3 M, 183.3 mmol, 2.1 eq) was slowly added dropwise to tetrahydrofuran (200 mL) containing N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide (16 g, 87.3 mmol, 1.0 eq) and stirred at -70°C for 1 hour. The reaction solution was slowly poured into water, stirred to room temperature, and then extracted with dichloromethane (300 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (EtOAc:PE = 0:100–10:90) to give a yellow solid 1-(1,5-dimethyl-1H-pyrazole-3-yl)-2,2-dimethylprop-1-one (7.5 g, yield 48%).

[0344] LCMS (ESI): m / z, 181.4 [M+H] +Rt = 1.38 min.

[0345] Step 3: Preparation of compound 1-(4-bromo-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one

[0346] N-bromosuccinimide (7.78 g, 43.6 mmol, 1.0 eq) was slowly added to acetonitrile (80 mL) containing 1-(1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one (7.5 g, 41.6 mmol, 1.0 eq) and stirred at 30 °C for 1 hour. The reaction solution was slowly poured into water and stirred to room temperature, then extracted with dichloromethane (300 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (EtOAc:PE = 0:100–10:90) to give the yellow solid 1-(4-bromo-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one (11.0 g, 92% yield).

[0347] 1 H NMR (400MHz, DMSO-d6) δ3.87(s,3H),2.26(s,3H),1.30(s,9H).

[0348] Step 4: Preparation of Compound 7

[0349] Compound 1-(4-bromo-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one (8 g, 30.8 mmol, 1 eq) and compound potassium ethylenetrifluoroborate (5.0 g, 37.0 mmol, 1.2 eq) were dissolved in dioxane (80 mL), and potassium carbonate (8.53 g, 61.7 mmol, 2.0 eq) and 1,1'-bis(diphenylphosphine)ferrocene palladium(II) dichloride (1.13 g, 1.5 mmol, 0.1 eq) were added. The reaction mixture was stirred at 100 °C for 16 hours. The reaction solution was slowly poured into water and stirred to room temperature, then extracted with dichloromethane (100 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product (EtOAc:PE = 0:100-10:90) was purified by column chromatography to give a brown solid compound 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one (3.12 g, yield 49%).

[0350] LCMS(ESI): m / z, 207.4 [M+H] + Rt = 1.53 min.

[0351] 1 H NMR (400MHz, DMSO-d6) δ7.08–6.96(m,1H),5.32–5.19(m,2H),3.84(d,J=9.0Hz,3H),2.35(d,J=12.1Hz,3H),1.45–1.35(m,9H).

[0352] Step 5: Preparation of Compound 8

[0353] Compound 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one (1.56 g, 7.5 mmol, 1 eq) was dissolved in acetonitrile (20 mL), and iodophenyl diacetic acid (2.57 g, 7.9 mmol, 1.05 eq) was added. Sulfuric acid (1.6 mL, 5%) was added at room temperature, and the reaction mixture was stirred at 25 °C for 1 hour. The reaction solution was slowly poured into water, stirred to room temperature, and then extracted with dichloromethane (50 mL × 3). The separated organic phase was concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (EtOAc:PE = 0:100-10:90) to give the yellow oily compound 2-(1,5-dimethyl-3-pentanoyl-1H-pyrazol-4-yl)acetaldehyde (1.2 g, yield 71%).

[0354] LCMS(ESI): m / z, 223.2 [M+H] + Rt = 1.22 min.

[0355] Step 6: Preparation of Compound 9

[0356] 2-(1,5-dimethyl-3-pentanoyl-1H-pyrazol-4-yl)acetaldehyde (2.7 g, 12.1 mmol, 1 eq) was dissolved in ethanol (20 mL), and sodium borohydride (460 mg, 12.1 mmol, 1.05 eq) was added. The reaction mixture was stirred at 0 °C for 1 hour. The reaction solution was slowly poured into water and stirred to room temperature, then extracted with dichloromethane (50 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (EtOAc:PE = 0:100–10:90) to give a yellow solid compound 1-(4-(2-hydroxyethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylprop-1-one (1.24 g, yield 45.5%).

[0357] LCMS(ESI): m / z, 225.4 [M+H] + Rt = 1.13 min.

[0358] Step 7: Preparation of Compound 12

[0359] 6-Bromoimidazole[1,2-a]pyridin-3-carboxaldehyde (7 g, 31.1 mmol, 1 eq) was dissolved in methanol (20 mL), and methylamine hydrochloride (1.16 g, 37.3 mmol, 1.2 eq) was added. The reaction mixture was stirred at room temperature for 16 hours. Sodium borohydride (1.77 g, 46.6 mmol, 1.5 eq) was added in portions, and the mixture was stirred at room temperature for another hour. The reaction solution was then slowly poured into water and stirred until room temperature was reached. Dichloromethane (50 mL × 3) was then added for extraction. The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (EtOAc:PE = 0:100-10:90) to give a pale yellow solid compound 1-(6-bromoimidazole[1,2-a]pyridin-3-yl)-N-methylmethylamine (3000 mg, yield 40%).

[0360] LCMS(ESI): m / z, 240.4 [M+H] + Rt = 0.79 min.

[0361] Step 8: Preparation of Compound 13

[0362] 1-(6-bromoimidazole[1,2-a]pyridin-3-yl)-N-methylmethylamine (3000 mg, 12.4 mmol, 1 eq) was dissolved in dichloromethane (20 mL), and di-tert-butyl dicarbonate (3.25 g, 14.9 mmol, 1.2 eq) and triethylamine (2.52 g, 24.9 mmol, 2.0 eq) were added. The reaction mixture was stirred at 30 °C for 16 hours. The reaction solution was slowly poured into water and stirred to room temperature, then extracted with dichloromethane (50 mL × 3). The separated organic phase was concentrated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (EtOAc:PE = 0:100-10:90) to give a white solid compound tert-butyl((6-bromoimidazole[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate (2.5 g, yield 58%).

[0363] LCMS(ESI): m / z, 340.4 [M+H] + Rt = 0.96 min.

[0364] Step 9: Preparation of Compound 16

[0365] Compound (3,4-difluoro-2-hydroxyphenyl)boronic acid (450 mg, 2.5 mmol, 1 eq) and compound tert-butyl((6-bromoimidazole[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate (924 mg, 2.7 mmol, 1.05 eq) were dissolved in dioxane (80 mL), and potassium carbonate (715 mg, 5.1 mmol, 2.0 eq) and 1,1'-bis(diphenylphosphine)ferrocene palladium(II) dichloride (189 mg, 0.2 mmol, 0.1 eq) were added. The reaction mixture was stirred at 100 °C for 16 hours. The reaction solution was slowly poured into water and stirred to room temperature, and then extracted with dichloromethane (30 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product (EtOAc:PE = 0:100-10:90) was purified by column chromatography to give a yellow solid compound 16 (900 mg, yield 89%).

[0366] LCMS(ESI): m / z, 390.2 [M+H] + Rt = 1.02 min.

[0367] Step 10: Preparation of Compound 17

[0368] Compound 16 (200 mg, 0.5 mmol, 1 eq) was dissolved in dioxane (10 mL), and compound 1-(4-(2-hydroxyethyl)-1,5-dimethyl-1H-pyrazole-3-yl)-2,2-dimethylprop-1-one (138.24 mg, 0.6 mmol, 1.2 eq), triphenylphosphine (purchased from Sigma) (270 mg, 1.03 mmol, 2.0 eq), and DEAD (purchased from Leyan) (179 mg, 1.03 mmol, 2.0 eq) were added. The reaction mixture was stirred at 100 °C for 16 hours. The reaction solution was slowly poured into water and stirred to room temperature, then extracted with dichloromethane (30 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (EtOAc:PE = 0:100-70:30) to obtain the crude product. The crude product was further purified by HPLC column chromatography (column: JB-C235-04, Bonnasil-BS C18, 20×250mm, 8μm, mobile phase: A: water (0.1% TFA), B: ACN, within 30 min, B: 10%-60%). The product was then lyophilized to obtain a yellow solid compound 17 (200mg, yield 65.3%).

[0369] LCMS(ESI): m / z, 596.4 [M+H] + Rt = 1.79 min.

[0370] Step 11: Preparation of compound Pl-R1dH

[0371] Compound 17 (40 mg, 0.068 mmol) was dissolved in dioxane (5 mL), and dioxane hydrochloride (0.3 mL, 4 M, 10 eq) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain crude PI-R1dH. The crude product was prepared by high performance liquid chromatography (column: JB-C235-04, Bonnasil-BS C18, 20 × 250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 14 min, B: 12%-42%). The eluent was lyophilized to give a white solid product PI-R1dH (8.0 mg, yield 32%).

[0372] LCMS(ESI): m / z, 496.4 [M+H] + Rt = 1.41 min.

[0373] 1 H NMR (400MHz, DMSO-d6) δ8.92(d,J=13.2Hz,3H),8.03(d,J=19.2Hz,1H),7.74(dd,J=24.2,10.4Hz,2H),7.38(dd,J=8.4,4 .1Hz,2H),4.68(s,2H),4.01(s,2H),3.74(s,4H),2.87(t,J=6.8Hz,2H),2.64(s,3H),2.03(d,J=2.0Hz,3H),1.21(s,9H).

[0374] Example 9: Preparation of compound PI-R1

[0375] Step 1: Preparation of compound PI-R1-b

[0376] Compound 17 (120 mg, 0.2 mmol, 1 eq) was dissolved in methanol (20 mL), and sodium borohydride (15.24 mg, 0.4 mmol, 1.05 eq) was added. The reaction mixture was stirred at 0 °C for 1 hour. The reaction solution was slowly poured into water and stirred until room temperature, then extracted with dichloromethane (50 mL × 3). The separated organic phases were concentrated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (EtOAc:PE = 0:100–10:90) to give a yellow solid compound PI-R1-b (110 mg, yield 91.3%).

[0377] LCMS(ESI): m / z, 598.4 [M+H] + Rt = 1.44 min.

[0378] Step 2: Preparation of compound PI-R1

[0379] At 25 °C, dioxane hydrochloride (1.2 mL, 4 M) was added to a solution of compound PI-R1-b (300 mg, 0.5 mmol) in dioxane (5 mL), and the mixture was stirred at 30 °C for 1 hour. The separated organic phase was concentrated under vacuum to obtain the crude product. The crude product was prepared by high performance liquid chromatography (column: BRIX-2860, Phenomenex Luna C18 50 × 250 mm, 10 μm; mobile phase: A: water (0.225% HCOOH), B: ACN, within 14 min, B: 12%–42%). The eluent was lyophilized to give a white solid product PI-R1 (200 mg, yield 80%).

[0380] LCMS(ESI): m / z, 496.2 [M+H] + Rt = 0.96 min.

[0381] 1 H NMR (400MHz, DMSO-d6) δ8.55(s,1H),7.57(d,J=9.6Hz,1H),7.50(s,1H),7.34(m,3H),5.37–4.62(m,1H),4.08(s,1 H),4.00(s,2H),3.89(m,2H),3.53(s,3H),2.81(m,1H),2.70–2.59(m,1H),2.25(s,3H),1.84(s,3H),0.76(s,9H).

[0382] Example 10 Preparation of compound PI-10-6Et

[0383] Step 1: Preparation of compound 10-6Et-c

[0384] At room temperature, fluorenemethyloxycarbonyl chloride (Fmoc-Cl) (3199.89 mg, 12.369 mmol, 3.0 eq) and triethylamine (1251.67 mg, 1.715 mL, 12.369 mmol, 3.0 eq) were added to a dichloromethane (45 mL) solution of compound 10-6MeiP-b (1400 mg, 4.123 mmol, 1.0 eq), nitrogen was purged, and the reaction was carried out at 45 °C for 18 h. The reaction solution was quenched with saturated sodium bicarbonate solution (50 mL), and the dichloromethane layer was collected. The aqueous layer was extracted twice with dichloromethane (50 mL * 2). The combined dichloromethane layers were washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1 / 5, 100 mL / min, 30 min) to give a yellow oily product compound 10-6Et-c (1600 mg, yield: 69.0%).

[0385] LCMS(ESI): m / z, 562.2 [M+H] + Rt = 1.80 min.

[0386] Step 2: Preparation of compound 10-6Et-d

[0387] The compound 10-6Et-c (1500 mg, 2.670 mmol, 1.0 eq) was added to a tetrahydrofuran (30 mL) solution with triethylamine trifluoroate (3984.17 mg, 26.700 mmol, 10.0 eq), and stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated sodium bicarbonate solution (100 mL) and extracted twice with ethyl acetate (100 mL * 2). The combined ethyl acetate layers were washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether = 0–30% (ethyl acetate gradient), 80 mL / min, 25 min) to give the yellow oily product compound 10-6Et-d (750 mg, yield: 62.0%).

[0388] LCMS(ESI): m / z, 448.4 [M+H] + Rt = 1.79 min.

[0389] Step 3: Preparation of compound 10-6Et-e

[0390] A solution of compound 10-6Et-d (750 mg, 1.676 mmol, 1.0 eq) and compound 16 (652.58 mg, 1.676 mmol, 1.0 eq) in dioxane (15 mL) was mixed with triphenylphosphine (1318.65 mg, 5.028 mmol, 3.0 eq) and DEAD (875.55 mg, 5.028 mmol, 3.0 eq), purged with nitrogen, and stirred at 100 °C for 3 hours. The reaction mixture was quenched with water (60 mL) and extracted twice with ethyl acetate (50 mL x 2). The combined ethyl acetate layers were washed with saturated brine (50 mL) and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 0–5% (methanol gradient), 60 mL / min, 35 min) to give the brown solid product compound 10-6Et-e (600 mg, yield: 60.0%).

[0391] LCMS (ESI): m / z, 596.8 [M+H] + Rt = 2.09 min.

[0392] 1H NMR(400MHz, CDCl3)δ8.55(s,1H),7.72–7.43(m,3H),7.09–7.03(m,1H),6.92(d,J=7.4Hz,1H),4.76(s,2H),4.12(q,J=7.0Hz,1H),4.0 5(t,J=5.6Hz,2H),3.73(s,3H),3.66–3.56(m,1H),3.27(d,J=5.2Hz,2H),2.74(s,3H),2.10(s,3H),1.45(s,9H),1.14(d,J=7.0Hz,6H).

[0393] Step 4: Preparation of compound 10-6Et-f

[0394] At room temperature, compound 10⁻⁶Et-e (160 mg, 0.268 mmol, 1.0 eq), acetaldehyde solution (0.536 mL, 5 M in THF, 10.0 eq), and acetic acid (79.16 mg, 1.341 mmol, 5.0 eq) in ethanol (5.0 mL) were stirred for 20 min, followed by the addition of NaBH₃CN (84.25 mg, 1.341 mmol, 5.0 eq). The reaction was stirred for 2 h, and LC-MS showed complete reaction. Water (20 mL) was added to the reaction solution at room temperature, followed by extraction with ethyl acetate (20 mL × 3). The organic phases were combined, washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / dichloromethane = 1 / 10 to 1 / 1) to give a yellow oily compound 10⁻⁶Et-f (100 mg, yield 59.2%).

[0395] LCMS (ESI): m / z, 625.5 [M+H] + .

[0396] Step 5: Preparation of compound 10⁻⁶Et-g

[0397] At room temperature, NaBH4 (60 mg, 1.601 mmol, 10 eq) was added to a MeOH (3 mL) solution of compound 10⁻⁶Et-f (100 mg, 0.160 mmol, 1.0 eq), and the mixture was stirred at 50 °C for 1 hour. LC-MS showed that the reaction was complete. Two drops of 1N HCl solution were added dropwise to the reaction mixture at room temperature, and the mixture was diluted with water (15 mL). The mixture was extracted twice with dichloromethane (15 mL), and the organic phases were combined. The mixture was washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. A crude product of the yellow oil compound 10⁻⁶Et-g (76 mg, yield 88.3%) was obtained.

[0398] LCMS(ESI): m / z, 627.4[(M+H)] + .

[0399] Step 6: Preparation of compound PI-10-6Et

[0400] At room temperature, compound 10⁻⁶Et-g (70 mg, 0.067 mmol, 1.0 eq) was added to 4 M dioxane hydrochloride (3.0 mL) and stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was purified by high performance liquid chromatography (JB-C235-04 column, Bonnasil-BS C18, 20*250 mm, 8 μm) with mobile phase A: water (0.1% TFA) and mobile phase B: acetonitrile, with acetonitrile elution ratios ranging from 20% to 60% to obtain a white solid product PI-10⁻⁶Et (39.20 mg, yield 91.4%).

[0401] PI-10-6Et:1H NMR (400MHz, DMSO-d6) δ8.94(d,J=9.8Hz,2H),8.09(s,1H),7.88(s,2H),7.39(dd,J=8.3,5.2Hz,2H),4.70(s,2H),4.00(dd,J=9.8,6.6Hz,4H),3.59(s ,3H),3.17(d,J=5.9Hz,2H),2.88(d,J=6.8Hz,2H),2.65(s,3H),2.12–1.84 (m, 4H), 0.87 (d, J = 6.6Hz, 3H), 0.77 (t, J = 7.8Hz, 3H), 0.63 (d, J = 6.8Hz, 3H).

[0402] Example 11 Preparation of compound PI-10-6Pr

[0403] Step 1: Preparation of compound 10-6Pr-f

[0404] At room temperature, acetic acid (331 mg, 0.316 mL, 5.614 mmol, 5.0 eq) and propionaldehyde (652.17 mg, 11.229 mmol, 10.0 eq) were added to 50.0 mL of ethanol containing 10-6Et-e (670 mg, 1.123 mmol, 1.0 eq) and NaBH3CN (352.81 mg, 5.614 mmol, 5.0 eq), purging with nitrogen, and the mixture was stirred in an oil bath at 50 °C for 3 hours. The reaction solution was quenched with 50 mL of saturated sodium bicarbonate solution and concentrated under reduced pressure to obtain a crude solution. The crude solution was then added to 50 mL of saturated sodium bicarbonate solution and extracted twice with ethyl acetate (50 mL x 2). The combined ethyl acetate layers were washed with 50 mL of saturated brine and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (CH3OH:CH2Cl2=1 / 20, 60mL / min, 25min) to give a yellow solid product compound 10-6Pr-f (650mg, yield 90.1%).

[0405] LCMS (ESI): m / z, 639.3 [M+H] + Rt = 1.39 min.

[0406] Step 2: Preparation of compound 10-6Pr-g

[0407] Compound 10⁻⁶Pr-f (650 mg, 1.018 mmol, 1.0 eq) was dissolved in methanol (10.0 mL) at room temperature, and sodium borohydride (385 mg, 10.176 mmol, 10.0 eq) was added. The reaction mixture was reacted at 50 °C for two hours. The reaction mixture was quenched with water (50 mL) and extracted three times with ethyl acetate (50 mL x 3). The combined ethyl acetate layers were washed with saturated brine (50 mL) and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to give the crude yellow solid compound 10⁻⁶Pr-g.

[0408] LCMS(ESI): m / z, 641.3[(M+H)] + Rt = 1.33 min.

[0409] Step 3: Preparation of compound PI-10-6Pr

[0410] At room temperature, compound 10⁻⁶Pr-g (652 mg, 1.018 mmol, 1.0 eq) was dissolved in dioxane (10 mL), and a 1,4-dioxane solution of 4 M HCl (5 mL) was added. The mixture was reacted at room temperature for one hour. LC-MS showed that the starting material reacted completely. The reaction solution was concentrated under reduced pressure to obtain the crude product. Approximately 20% of the crude product was purified by high-performance liquid chromatography (HPLC) (JB-C235-04 column, Bonnasil-BS C18, 30*150 mm, 5 μm) with mobile phase A: water (0.1% TFA) and mobile phase B: acetonitrile, with acetonitrile elution ratios ranging from 19% to 33%, 25 mL / min, 30 min) to obtain the yellow solid product PI-10⁻⁶Pr (55.13 mg).

[0411] PI-10-6Pr:LCMS(ESI):m / z,541.4[M+H] + Rt = 0.97 min.

[0412] 1H NMR(400MHz,DMSO-d6)δ9.06(s,2H),9.01(s,1H),8.18(s,1H),7.99–7.89 (m,2H),7.44–7.33(m,2H),4.73(s,2H),4.04–3.95(m,3H),3.58(s,3H),3. 24–3.09(m,2H),2.84–2.73(m,2H),2.66(s,3H),2.01(s,3H),1.21–1.11( m, 2H), 0.88 (d, J = 6.6Hz, 3H), 0.69 (t, J = 7.4Hz, 3H), 0.63 (d, J = 6.6Hz, 3H).

[0413] Example 12 Preparation of compound PI-10-6Bu

[0414] Step 1: Preparation of compound 10-6Bu-c

[0415] Acetic acid (0.828 mL, 14.725 mmol) was added to ethanol (50.0 mL) containing compound 10-6MeiP-b (1.00 g, 2.945 mmol, 1.0 eq) and butyraldehyde (2.193 mL, 29.45 mmol, 10.0 eq), and the mixture was stirred at 25 °C for 20 min. Then, NaBH3CN (0.93 g, 14.725 mmol) was added to the mixture, and the mixture was stirred at 25 °C for 2 h. LC-MS showed that the reaction was complete. The mixture was quenched with NH4Cl (20 mL), and the solvent was removed under reduced pressure. The mixture was extracted with EtOAc (30 mL × 3). The organic phase was washed with brine, dried over anhydrous Na2SO4, and the solvent was removed under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (EtOAc:PE = 0–30% (EtOAc gradient), 60 mL / min, 25 min) to obtain compound 10⁻⁶Bu⁻c (800 mg, yield: 68%), which was a colorless oil.

[0416] LCMS(ESI): m / z, 396.8[(M+H)] + Rt = 2.856 min.

[0417] Step 2: Preparation of compound 10-6Bu-d

[0418] HF / Py (3259.59 mg, 20.219 mmol) was added to a THF (10 mL) solution of compound 10⁻⁶Bu⁻c (800 mg, 2.022 mmol) at 25 °C, and the mixture was stirred at 25 °C for 18 hours. LC-MS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (MeOH:DCM = 0–5% (MeOH gradient), 60 mL / min, 25 min) to give a colorless oily substance 10⁻⁶Bu⁻d (420 mg, yield: 74%).

[0419] LCMS(ESI): m / z, 282.6[(M+H)] + Rt = 1.755 min.

[0420] Step 3: Preparation of compound 10-6Bu-e

[0421] DEAD (594.15 mg, 3.412 mmol) was added to a solution of compound 10-6Bu-d (320 mg, 1.137 mmol), compound 16 (442.82 mg, 1.137 mmol), and Ph3P (894.81 mg, 3.412 mmol) in 20 mL of Dioxane at 0 °C under nitrogen atmosphere. The mixture was then stirred at 100 °C for 18 hours. LC-MS showed that the reaction was complete. The reaction mixture was diluted with 50 mL of H2O and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (MeOH:DCM = 0–3% (MeOH gradient), 60 mL / min, 25 min) to give compound 10-6Bu-e (1.62 g, yield: 98%) as a brown oil.

[0422] LCMS(ESI): m / z, 654.0 [M+H] + Rt = 2.783 min.

[0423] Step 4: Preparation of compound 10-6Bu-f

[0424] At 0 °C, under nitrogen atmosphere, NaBH4 (5.00 g, 132.164 mmol) was added to a MeOH (20 mL) solution of compound 10⁻⁶Bu⁻e (1.42 g, 0.979 mmol). The resulting mixture was stirred at 50 °C under N₂ for 2 hours. LC-MS showed that the reaction was complete. The mixture was quenched with NH₄Cl (10 mL). The mixture was extracted with EtOAc (20 mL × 3). The organic phase was washed with brine, dried over anhydrous Na₂SO₄, and the solvent was removed under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (MeOH:DCM = 0–6% (MeOH gradient), 60 mL / min, 25 min) to give compound 10⁻⁶Bu⁻f (660 mg, yield: 99%) as a brown oil.

[0425] LCMS(ESI): m / z, 655.5 [M+H] + Rt = 2.290 min.

[0426] Step 5: Preparation of compound PI-10-6Bu

[0427] At 25 °C, HCl / dioxane (4 M) (1 mL) was added to compound 10⁻⁶Bu-f (50.00 mg, 0.076 mmol). The resulting mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give the crude product. The residue was purified by preparative HPLC (JB-C235-04 column, Bonnasil-BS C18, 20*250 mm, 8 μm) with mobile phase A: water (0.1% TFA) and mobile phase B: acetonitrile, with acetonitrile elution ratios ranging from 10% to 45% to give PI-10⁻⁶Bu (20.97 mg, yield: 49%) as a white solid.

[0428] LCMS(ESI): m / z, 555.5 [M+H] + Rt = 1.933 min.

[0429] 1H NMR (400MHz, CD3OD) δ9.00(s,1H),8.27(s,1H),8.11(dd,J=9.4,1.5Hz,1H),7.96(dd,J=9.4,0.7H z,1H),7.44–7.35(m,1H),7.22(dt,J=16.5,8.3Hz,1H),4.84(s,2H),4.53(d,J=5.4Hz,1H),4.14( t,J=5.4Hz,2H),3.74(s,3H),3.56(s,2H),3.30–3.22(m,2H),2.86(s,3H),2.29(s,3H),2.14(dq, J=13.4,6.7Hz,1H),1.44(dt,J=15.3,7.7Hz,2H),1.35–1.25(m,2H),0.88(dd,J=9.9,4.7Hz,9H).

[0430] Example 13 Preparation of compound PI-10-6

[0431] Step 1: Preparation of compound 10-6-b

[0432] At 0 °C, a solution of compound 10⁻⁶-S (5.00 g, 45.0 mmol) in dichloromethane (80 mL) was successively treated with TEA (13.7 g, 135 mmol), DMAP (0.55 g, 4.50 mmol), and (BOC)₂O (11.8 g, 54.0 mmol). The mixture was stirred at 25 °C for 16 hours. LC-MS showed that the reaction was complete. The mixture was concentrated to give a crude product. The residue was purified by silica gel column chromatography (SiO₂, PE / EA = 10 / 1 to 1 / 2) to give a colorless oily 10⁻⁶-b (3.55 g, yield: 36.0%).

[0433] LCMS(ESI): m / z, 211.6 [M+H] + Rt = 1.13 min.

[0434] Step 2: Preparation of compound 10-6-c

[0435] NaH (1.08 g, 26.9 mmol) was slowly added to a DMF (50 mL) solution of 10⁻⁶-b (3.55 g, 16.8 mmol) at 0 °C. The mixture was stirred at 0 °C for 30 min. Then, 2-bromoethoxytert-butyldiphenylsilane (7.94 g, 21.8 mmol) was added, and the reaction mixture was stirred at 25 °C for 4 h. LC-MS showed that the reaction was complete; the reaction mixture was quenched with saturated NH₄Cl at 0 °C, diluted with 50 mL H₂O, and extracted with ethyl acetate (60 mL × 3). The combined organic layers were washed with brine (80 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (SiO₂, PE / EA = 10 / 1 to 1 / 1) to give a yellow oily 10⁻⁶-c (5.90 g, yield: 71%).

[0436] LCMS(ESI): m / z, 493.5 [M+H] + Rt = 1.98 min.

[0437] Step 3: Preparation of compound 10-6-d

[0438] Compound 10⁻⁶-c (5.70 g, 11.5 mmol) was dissolved in TFA (5 mL) and DCM (50 mL), and the reaction mixture was stirred at 25 °C for 16 hours. LC-MS showed that the reaction was complete. The reaction mixture was quenched with saturated NaHCO₃, diluted with H₂O, and then extracted with DCM (120 mL × 3). The combined organic layers were washed with saturated brine (120 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give a yellow oily substance 10⁻⁶-d (4.15 g, yield: 91%).

[0439] LCMS (ESI): m / z, 393.6 [M+H] + Rt = 1.35 min.

[0440] Step 4: Preparation of compound 10-6-e

[0441] Potassium carbonate (2.91 g, 21.1 mmol) and CH3I (1.50 g, 10.5 mmol) were added sequentially to a DMF (40 mL) solution of 10⁻⁶-d (4.15 g, 10.5 mmol); the reaction mixture was stirred at 25 °C for 16 hours. LC-MS showed that the reaction was complete. The mixture was quenched with H₂O (50 mL) and extracted with EtOAc (80 mL × 3). The EtOAc phase was washed with brine, dried over Na₂SO₄, and subjected to reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 1 / 10 to 1 / 3) to give 10⁻⁶-e (3.2 g, yield: 74%).

[0442] LCMS (ESI): m / z, 407.6 [M+H] + Rt = 1.44 min.

[0443] Step 5: Preparation of compound 10-6-f

[0444] At room temperature, NBS (1.40 g, 7.85 mmol) was added to a chloroform (340 mL) solution of compound 10-6-e (3.2 g, 7.85 mmol), and the reaction mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The mixture was quenched with water (60 mL) and extracted with DCM (60 mL × 2). The combined organic layers were washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EtOAc = 10:1) to give a yellow oily 10-6-f (2.2 g, yield: 57%).

[0445] LCMS (ESI): m / z, 485.4 [M+H] + Rt = 2.08 min.

[0446] Step 6: Preparation of compound 10⁻⁶-g

[0447] Under nitrogen protection at -78°C, n-butyllithium (0.987 mL, 2.47 mmol, 2.5 M) was slowly added to a 20 mL solution of 10⁻⁶-f (1.00 g, 2.06 mmol) in diethyl ether. The reaction mixture was stirred at -78°C for half an hour, followed by the slow addition of DMF (0.23 g, 3.08 mmol, 0.237 mL), and the mixture was stirred at -78°C for 2 hours. LC-MS showed that the reaction was complete. The reaction mixture was heated to 0°C, and the reaction was quenched by the slow addition of HCl (4 M). The mixture was then diluted with H₂O (50 mL) and extracted with ethyl acetate (60 mL × 3). The combined organic layers were washed with brine (80 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (SiO2, PE / EA = 50 / 1 to 5 / 1) to give 10⁻⁶-g (750 mg, purity: 65%, yield: 54%) as a yellow oil.

[0448] LCMS(ESI): m / z, 494.4 [M+H] + , Rt = 1.89 min.

[0449] Step 7: Preparation of compound 10⁻⁶-h

[0450] At 0 °C, tert-butylmagnesium chloride (0.987 mL, 1.68 mmol, 1.7 M) was slowly added to a 10⁻⁶ g (750 mg, 1.12 mmol, 65% purity) solution of THF (10 mL). The reaction mixture was stirred at 0 °C for 2 h. LC-MS showed that the reaction was complete; the reaction mixture was quenched at 0 °C with saturated NH₄Cl, diluted with H₂O (30 mL), and extracted with ethyl acetate (40 mL × 3). The combined organic layers were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by passing it through a C18 reversed-phase column (mobile phase A: 0.1% TFA, mobile phase B: acetonitrile, B: 5-50%) to give 500 mg of 10⁻⁶-h (purity: 60%, yield: 54%) as a yellow oil.

[0451] LCMS(ESI): m / z, 494.4 [M+H] + , Rt = 1.89 min.

[0452] Step 8: Preparation of compound 10-6-i

[0453] At room temperature, triethylamine trihydrofluoric acid (979 mg, 6.08 mmol) was added to a 5 mL THF solution of compound 10⁻⁶-h (500 mg, purity: 60%, 0.608 mmol), and the mixture was stirred at 25 °C for 2 hours. LC-MS showed that the reaction was complete. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / MeOH = 100:1 to 92:8) to give a brown oily compound 10⁻⁶-i (150 mg, yield: 96%).

[0454] LCMS (ESI): m / z, 255.6 [M+H] + Rt = 0.81 min.

[0455] Step 9: Preparation of compound 10-6-j

[0456] Compound 16 (183 mg, 0.470 mmol), triphenylphosphine (370 mg, 1.41 mmol), and DEAD (246 mg, 1.41 mmol) were added sequentially to a 1,4-dioxane (5 mL) solution of compound 10-6-i (120 mg, 0.470 mmol). The reaction mixture was stirred at 100 °C under a nitrogen atmosphere for 4 hours. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by C18 column chromatography (mobile phase A: 0.1% TFA / H2O, mobile phase B: acetonitrile, B: 5-50%) to give a yellow solid 10-6-j (55.0 mg, purity: 80%, yield: 15%).

[0457] LCMS (ESI): m / z, 627.4 [M+H] + Rt = 1.22 min.

[0458] Step 10: Preparation of compound PI-10-6

[0459] Compound 10⁻⁶-j (30.0 mg, purity: 80%, 0.038 mmol) was dissolved in a mixed solution of DCM (3 mL) and TFA (1 mL) at 0 °C. The reaction solution was stirred at 25 °C for 2 hours. LC-MS showed that the reaction was complete. The solution was removed by concentration under reduced pressure, and the crude product was purified by high performance liquid chromatography (flow rate: 25 mL / min, column: Ultimate XB-C18, 30*150 mm, 5 μm, mobile phase A: 0.1% / water, mobile phase B: acetonitrile, B: 15-35%) to give a yellow solid PI-10⁻⁶ (19.0 mg, yield: 94%).

[0460] PI-10-6: LCMS(ESI): m / z, 527.4[M+H] + Rt = 0.99 min.

[0461] 1H NMR (400MHz, CD3OD) δ9.01(s,1H),8.27(s,1H),8.13(dd,J=9.2,1.2Hz,1H),7.98(d,J=9.2Hz,1H),7.44–7.36(m,1H),7.23(dd,J=16.8,9.2Hz,1H ),4.84(s,2H),4.44(s,1H),4.23–4.15(m,1H),4.14–4.06(m,1H),3.73( s,3H),3.54(s,2H),3.01(s,3H),2.85(s,3H),2.32(s,3H),0.91(s,9H).

[0462] Example 14 Preparation of compound PI-10-6Ett

[0463] Step 1: Preparation of compound 10-6Ett-e

[0464] Potassium carbonate (2.91 g, 21.1 mmol) and iodoethane (1.64 g, 10.5 mmol) were added sequentially to a DMF (40 mL) solution of 10⁻⁶-d (4.15 g, 10.5 mmol); the reaction mixture was stirred at 25 °C for 22 h. LC-MS showed that the reaction was complete. The mixture was quenched with H₂O (50 mL) and extracted with EtOAc (80 mL × 3). The EtOAc phase was washed with brine, dried over Na₂SO₄, and subjected to reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 1 / 10 to 1 / 3) to give 10⁻⁶Ett-e (3.6 g, yield: 81%).

[0465] LCMS (ESI): m / z, 422.3 [M+H] + Rt = 1.44 min.

[0466] Step 2: Preparation of compound 10-6Ett-f

[0467] At room temperature, NBS (1.52 g, 8.53 mmol) was added to a chloroform (360 mL) solution of compound 10⁻⁶Ett-e (3.6 g, 8.53 mmol), and the reaction mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The mixture was quenched with water (60 mL) and extracted with DCM (60 mL × 2). The combined organic layers were washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography (PE:EtOAc = 10:1) to give a yellow oily 10⁻⁶Ett-f (2.3 g, yield: 53%).

[0468] LCMS (ESI): m / z, 500.2 [M+H] + Rt = 2.10 min.

[0469] Step 3: Preparation of compound 10-6Ett-g

[0470] Under nitrogen protection at -78°C, n-butyllithium (2.2 mL, 5.52 mmol, 2.5 M) was slowly added to a solution of 10⁻⁶ Ett-f (2.3 g, 4.6 mmol) in 45 mL of diethyl ether. The reaction mixture was stirred at -78°C for half an hour, followed by the slow addition of DMF (0.46 g, 6.16 mmol, 0.474 mL), and the mixture was stirred at -78°C for 2.5 hours. LC-MS showed that the reaction was complete. The reaction mixture was heated to 0°C, and the reaction was quenched by the slow addition of HCl (4 M). The mixture was then diluted with H₂O (120 mL) and extracted with ethyl acetate (120 mL × 3). The combined organic layers were washed with brine (120 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (SiO2, PE / EA = 50 / 1 to 5 / 1) to give a yellow oily 10⁻⁶Ett-g (1.63 g, purity: 70%, yield: 55%).

[0471] LCMS (ESI): m / z, 450.3 [M+H] + Rt = 1.91 min.

[0472] Step 4: Preparation of compound 10-6Ett-h

[0473] At 0 °C, tert-butylmagnesium chloride (1.48 mL, 2.52 mmol, 1.7 M) was slowly added to a solution of 10⁻⁶ Ett⁻ ...

[0474] LCMS (ESI): m / z, 508.3 [M+H] + Rt = 1.90 min.

[0475] Step 5: Preparation of compound 10-6Ett-i

[0476] At room temperature, triethylamine trihydrofluoric acid (2.12 g, 13.1 mmol) was added to a THF (5 mL) solution of compound 10⁻⁶Ett-h (1.11 g, purity: 60%, 1.31 mmol), and the mixture was stirred at 25 °C for 2.5 h. LC-MS showed that the reaction was complete. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (DCM / MeOH = 100:1 to 92:8) to give a brown oily substance 10⁻⁶Ett-i (325 mg, yield: 97%).

[0477] LCMS (ESI): m / z, 255.2 [M+H] + Rt = 0.83 min.

[0478] Step 6: Preparation of compound 10-6Ett-j

[0479] Compound 16 (494 mg, 1.27 mmol), triphenylphosphine (999 mg, 3.81 mmol), and DEAD (664 mg, 3.81 mmol) were added sequentially to a 1,4-dioxane (12 mL) solution of compound 10-6Ett-i (325 mg, 1.27 mmol). The reaction mixture was stirred at 100 °C under a nitrogen atmosphere for 4 hours. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by C18 column chromatography (mobile phase A: 0.1% TFA / H2O, mobile phase B: acetonitrile, B: 5-50%) to give a yellow solid 10-6Ett-j (169 mg, purity: 82%, yield: 17%).

[0480] LCMS (ESI): m / z, 641.4 [M+H] + Rt = 1.24 min.

[0481] Step 7: Preparation of compound PI-10-6Ett

[0482] Compound 10-6Ett-j (169 mg, purity: 82%, 0.22 mmol) was dissolved in a mixed solution of DCM (6 mL) and TFA (2 mL) at 0 °C. The reaction mixture was stirred at 25 °C for 2 hours. LC-MS showed that the reaction was complete. The solution was removed by concentration under reduced pressure, and the crude product was purified by high performance liquid chromatography (flow rate: 25 mL / min, column: Ultimate XB-C18, 30*150 mm, 5 μm, mobile phase A: 0.1% / water, mobile phase B: acetonitrile, B: 15-35%) to give a yellow solid PI-10-6Ett (107 mg, yield: 94%).

[0483] PI-10-6Ett: LCMS(ESI): m / z, 541.3[M+H] +, Rt=1.01min.

[0484] 1 H NMR (400MHz, CD3OD) δ9.01 (s, 1H), 8.26 (s, 1H), 8.11 (dd, J = 9.2, 1.2Hz, 1H), 7.9 8(d,J=9.2Hz,1H),7.45–7.35(m,1H),7.23(dd,J=16.8,9.2Hz,1H),4.84(s,2H), 4.44(s,1H),4.22–4.15(m,1H),4.14–4.05(m,1H),3.71(s,3H),3.53(s,2H),2.9 9(s,3H),2.75(d,J=6.8Hz,2H),2.32(s,3H),0.91(s,9H),0.76(t,J=6.8Hz,3H).

[0485] Linker-payload preparation

[0486] Example 15: Synthesis of compound LPi-B1

[0487] Step 1: Preparation of compound LP-R1-a

[0488] To a solution of compound LP-R1-a-1 (5.0 g, 12.6 mmol, 1 eq) in ACN (125 mL), compounds LP-R1-a-2 (3.53 g, 21.2 mmol, 1.68 eq) and silver oxide (13.1 g, 105 mmol, 8.4 eq) were added. After the addition was complete, the mixture was stirred at 25 °C for 4 hours. LC-MS showed that the reaction was complete. The reaction mixture was filtered, and the filtrate was diluted with water (300 mL) and then extracted with EtOAc (400 mL × 3). The combined organic phases were washed with brine (300 mL), dried over anhydrous sodium sulfate, and filtered to obtain the filtrate. The filtrate was concentrated under reduced pressure to give the crude yellow solid product LP-R1-a (6.0 g, 90% purity, yield 89%).

[0489] LCMS (ESI): m / z, 506.0 [M+Na] + Rt = 1.14 min.

[0490] Step 2: Preparation of compound LP-R1-b

[0491] Compound LP-R1-a (6.00 g, 12.4 mmol, 1 eq) was dissolved in isopropanol (25 mL) and chloroform (25 mL). NaBH4 (700 mg, 18.5 mmol, 1.5 eq) was added at 0 °C. The reaction mixture was stirred at 25 °C for 2 hours, and LC-MS showed that the starting material reacted completely. The reaction mixture was filtered, and the filtrate was diluted with water (200 mL) and then extracted with DCM (300 mL × 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, and filtered to obtain the filtrate. The filtrate was concentrated under reduced pressure to obtain the crude yellow solid product LP-R1-b (4.10 g, 70% purity, yield 47.63%).

[0492] LCMS(ESI): m / z, 508.0 [M+Na] + , Rt = 1.05 min.

[0493] Step 3: Preparation of compound LP-R1-c

[0494] Platinum dioxide (0.21 g, 0.923 mmol, 0.32 eq) was added to a 20 mL solution of compound LP-R1-b (2.00 g, 2.88 mmol, 1 eq) in ethyl acetate at room temperature. The reaction mixture was stirred at 25 °C for 18 hours. LC-MS showed that the starting material reacted almost completely. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:2) to give a white solid product (1.30 g, 99% yield).

[0495] LCMS(ESI): m / z, 455.7 [M+H] + Rt = 1.226 min.

[0496] Step 4: Preparation of compound LP-R1-d

[0497] Compound LP-R1-c (100 mg, 0.220 mmol, 1 eq) was dissolved in DCM (2 mL) and MeOH (1 mL) solutions. LP-R1-c-1 (103 mg, 0.329 mmol, 1.5 eq) and EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, 109 mg, 0.439 mmol, 2 eq) were added. The reaction mixture was stirred at 25 °C for 18 hours, and LC-MS showed complete reaction of the starting material. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (PE:EtOAc = 0:1) to give 154 mg of a white solid product, yield 4.93%, purity 80%.

[0498] LCMS(ESI): m / z, 748.7 [M+H] +, Rt=1.551min.

[0499] Step 5: Preparation of compound LP-R1-e

[0500] Compound LP-R1-d (154 mg, 0.165 mmol, 1 eq) was dissolved in DMF (2 mL), and NPC (di(p-nitrobenzene) carbonate, 100 mg, 0.329 mmol, 2 eq) and DIEA (N,N-diisopropylethylamine, 42.5 mg, 0.329 mmol, 2 eq) were added. The reaction mixture was stirred at 25 °C for 2 hours, and LC-MS showed that the starting material reacted completely. The reaction mixture was filtered, and the filtrate was diluted with EtOAc (50 mL × 3), washed with water (50 mL), dried over the organic phase, filtered, and evaporated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 0:1) to give a yellow oily liquid product (120 mg, yield 80%).

[0501] LCMS(ESI): m / z, 913.4 [M+H] + Rt = 1.714 min.

[0502] Step 6: Preparation of compound LP-R1-f

[0503] Compound LP-R1-e (65 mg, 0.071 mmol, 1 eq) was dissolved in DMF (2 mL), and PI-R1 (89 mg, 0.178 mmol, 2.5 eq), HOBt (1-hydroxybenzotriazole, 10 mg, 0.071 mmol, 1 eq), and DIEA (28 mg, 0.213 mmol, 3 eq) were added. The reaction mixture was stirred at 25 °C for 1 hour, and LC-MS showed that the starting material reacted completely. The reaction mixture was filtered, and the filtrate was diluted with EtOAc (50 mL × 3), washed with water (50 mL), dried over the organic phase, filtered, and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give a yellow oily liquid product (84 mg, yield 93%).

[0504] LCMS(ESI): m / z, 1271.5 [M+H] + Rt = 1.547 min.

[0505] Step 7: Preparation of compound LP-R1-g

[0506] Compound LP-R1-f (40.0 mg, 0.031 mmol, 1 eq) was dissolved in acetonitrile (1.2 mL) and water (0.8 mL). 1 M NaOH (0.22 mL) was added at 0 °C, and the reaction mixture was stirred at 25 °C for 1 hour. LC-MS showed that the starting material reacted completely. 2 M HCl was added to the reaction mixture until pH = 5. The reaction mixture was filtered and evaporated to dryness. The residue was lyophilized to give a white solid product (35.0 mg, yield 61.17%, 50% purity).

[0507] LCMS(ESI): m / z, 910.3 [M+H] + Rt = 0.884 min.

[0508] Step 8: Preparation of compound LPi-B1

[0509] Compound LP-R1-g (35 mg, 0.038 mmol, 1 eq) was dissolved in DMF (1.5 mL), and LP-R1-g-1 (13 mg, 0.038 mmol, 1 eq) and DIEA (10 mg, 0.077 mmol, 2 eq) were added. The reaction mixture was stirred at 25 °C for 1 hour, and LCMS showed that the starting material reacted completely. The reaction mixture was concentrated, and the residue was prepared by high performance liquid chromatography to give a white solid product LPi-B1 (11.32 mg, yield 26%).

[0510] The preparation conditions for high performance liquid chromatography are as follows:

[0511] The preparative chromatogram was manufactured by Shimadzu, model JB-C235-04. The column was a Prime C18 30×150mm×5μm. The mobile phase was water (0.1% TFA)-ACN, with acetonitrile elution from 25% to 45% over 14 min.

[0512] LCMS(ESI): m / z, 1149.4 [M+H] + Rt = 1.138 min.

[0513] 1H NMR (400MHz, DMSO-d6) δ9.03(s,1H),8.93(s,1H),8.22-8.20(m,2H),8.00(d,J=9.2Hz,1H),7.9 2(d,J=8.2Hz,2H),7.34(dd,J=16.8,9.2Hz,1H),7.15(s,1H),7.01(s,2H),5.05(s,2H),4.94(d, J=8.4Hz,2H),4.77(d,J=7.4Hz,1H),4.00(s,1H),3.95-3.81(m,5H),3.54(dd,J=5.6,3.2Hz,23H ), 2.92 (s, 3H), 2.74 (s, 1H), 2.63-2.55 (m, 2H), 2.28 (t, J = 6.4Hz, 2H), 1.95 (s, 3H), 0.70 (s, 9H).

[0514] Example 16: Preparation of compound LPi-7-1

[0515] Step 1: Preparation of compound LPi-7-1-a

[0516] Compound LP-R1-e (52 mg, 0.046 mmol, 1 eq) was dissolved in DMF (4 mL), and PI-7-1 (20 mg, 0.038 mmol, 2.5 eq), HOBt (5.16 mg, 0.038 mmol, 1 eq), and DIEA (15 mg, 0.114 mmol, 3 eq) were added. The reaction mixture was stirred at 30 °C for 1 hour, and LC-MS showed that the starting material reacted completely. The reaction mixture was filtered, and the filtrate was diluted with EtOAc (50 mL × 3), washed with water (50 mL), dried over the organic phase, filtered, and evaporated to dryness. The residue was purified by silica gel column chromatography (DCM:MeOH = 5:95) to give a white solid product (24 mg, yield 48.40%).

[0517] LCMS(ESI): m / z, 1297.5 [M+H] + Rt = 1.455 min.

[0518] Step 2: Preparation of compound LPi-7-1-b

[0519] Compound LPi-7-1-a (24 mg, 0.018 mmol, 1 eq) was dissolved in acetonitrile (2.4 mL) and water (1.2 mL). 1 M NaOH (0.12 mL) was added at 0 °C, and the reaction mixture was stirred at 15 °C for 1 hour. LC-MS showed that the starting material reacted completely. 2 M HCl was added to the reaction mixture to pH 6-7. The reaction mixture was filtered and evaporated to dryness. The residue was purified by C18 chromatography (0.1% TFA, H2O:ACN = 59:41) to give a yellow solid compound LPi-7-1-b (11 mg, yield 63%).

[0520] LCMS(ESI): m / z, 935.6 [M] + Rt = 1.025 min.

[0521] Step 3: Preparation of compound LPi-7-1

[0522] Compound LPi-7-1-b (11 mg, 0.012 mmol, 1 eq) was dissolved in DMF (2 mL), and LP-R1-g-1 (commercially available, 6 mg, 0.018 mmol, 1.5 eq) and DIEA (3 mg, 0.024 mmol, 2 eq) were added. The reaction mixture was stirred at 15 °C for 1 hour, and LC-MS showed that the starting material reacted completely. The reaction mixture was concentrated, and the residue was prepared by high performance liquid chromatography (column: JB-C235-04, Bonnasil-BS C18, 20 × 250 mm, 8 μm; mobile phase: A: water (0.1% TFA), B: ACN, within 14 min, B: 21%–41%) to give a white solid product LPi-7-1 (5.23 mg, yield 38%).

[0523] LCMS(ESI): m / z, 1175.4 [M+1] + Rt = 1.32 min.

[0524] Example 17: Preparation of compound LPi-10-6Et

[0525] Step 1: Preparation of compound LPi-10-6Et-a

[0526] Ethyl diisopropylamine (0.362 mL, 2.184 mmol) was added in a single addition to a DMF (5 mL) solution of PI-10-6Et (230 mg, 0.437 mmol) at 0 °C. LP-R1-e (379.02 mg, 0.437 mmol) and HOBt (59.01 mg, 0.437 mmol) were then added. The mixture was stirred at 25 °C for 30 min. LC-MS showed the reaction was complete. The reaction mixture was quenched with water (20 mL) at 0 °C. The resulting mixture was extracted with EtOAc (20 mL × 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was purified by rapid silica gel column chromatography, eluting with 8% MeOH:DCM, to give compound LPi-10-6Et-a (300 mg, 0.231 mmol, 52.78%) as a white solid.

[0527] LCMS(ESI): m / z, 1301.5 [M+H] + Rt = 1.598 min.

[0528] Step 2: Preparation of compound LPi-10-6Et-b

[0529] Compound LPi-10-6Et-a (300 mg, 0.231 mmol, 1 eq) was dissolved in acetonitrile (7.5 mL) and water (5.0 mL). NaOH (73.76 mg, 1.844 mmol) was slowly added at 0 °C, and the mixture was stirred at 25 °C for 1 hour. LC-MS showed the reaction was complete. The solution was concentrated under reduced pressure to give the crude product. The residue was purified by preparative high-performance liquid chromatography (20-70% ACN / H₂O, 0.1% TFA) to give compound LPi-10-6Et-b (200 mg, 0.213 mmol, 92.40%) as a white solid.

[0530] LCMS(ESI): m / z, 939.5 [M] + Rt = 1.132 min.

[0531] Step 3: Preparation of compound LPi-10-6Et

[0532] Ethyl diisopropylamine (0.095 mL, 0.575 mmol) was added to a DMF (2 mL) solution of compound LPi-10-6Et-b (180 mg, 0.192 mmol) and compound LP-R1-g-1 (169.80 mg, 0.479 mmol) at 0 °C. The mixture was then stirred at 25 °C for 10 min. LCMS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give the crude product. The residue was purified by preparative high performance liquid chromatography (0-100% ACN / H2O, 0.1% TFA) to give compound LPi-10-6Et (137.92 mg, 0.103 mmol, 53.82%) as a white solid.

[0533] LPI-10-6Et: LCMS(ESI): m / z, 1179.7[M+1] + , Rt=1.194min.

[0534] 1 H NMR(400MHz,MeOD)δ9.02(s,1H),8.20(d,J=8.6Hz,1H),8.10(t,J=4.0Hz,2H),7.96(dt,J=14.4,5.6Hz,1H),7.08(s,2H),6.98(s,1H),6.91–6.8 2(m,1H),6.78(d,J=4.3Hz,2H),5.17(s,2H),5.00(s,2H),4.90–4.87(m, 4H),4.83(d,J=1.6Hz,4H),4.38(d,J=30.4Hz,1H),4.13(s,2H),3.82(dd ,J=26.0,8.4Hz,1H),3.72–3.64(m,5H),3.61(dt,J=10.0,3.1Hz,3H),3. 56–3.41(m,12H),3.20(s,1H),3.13(dt,J=3.4,1.7Hz,1H),3.04(s,3H), 2.55(s,2H),2.41(t,J=6.1Hz,2H),2.20(s,3H),2.12–2.02(m,1H),1.29 (s,1H),0.97(s,3H),0.84(dd,J=6.6,1.3Hz,3H),0.78(t,J=8.3Hz,3H).

[0535] Example 18: Preparation of compound LPi-10-6Pr

[0536] Step 1: Preparation of compound LPi-10-6Pr-a

[0537] At room temperature, ethyl diisopropylamine (657.40 mg, 5.086 mmol, 5.0 eq) and HOBT (137.46 mg, 1.017 mmol, 1.0 eq) were added to a DMF (10.0 mL) solution of crude PI-10-6Pr (520 mg, 0.961 mmol) and compound LP-R1-e (929.63 mg, 1.017 mmol, 1.0 eq), and the mixture was reacted at room temperature for one hour. The reaction solution was quenched with water (50 mL) and extracted three times with ethyl acetate (50 mL × 3). The combined ethyl acetate layers were washed with saturated brine (50 mL) and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 1 / 15, 50 mL / min, 40 min) to give the yellow solid product compound LPi-10-6Pr-a (1580 mg, purity: 80%, yield: 94%).

[0538] LCMS(ESI): m / z, 1315.6 [M+H] + Rt = 1.498 min.

[0539] Step 2: Preparation of compound LPi-10-6Pr-b

[0540] A solution of compound LPi-10-6Pr-a (1580 mg, purity: 80%, 0.961 mmol, 1 eq) in acetonitrile (24.0 mL) / H2O (15.0 mL) was added to an aqueous solution of 1N NaOH (7.7 mL, 7.687 mmol, 8.0 eq) under ice bath conditions, and the mixture was stirred at room temperature for one hour. The pH of the reaction solution was adjusted to 3 with formic acid, and then purified by C18 reversed-phase column chromatography (mobile phase A: H2O (0.1% TFA), mobile phase B: acetonitrile, B: 5–50%, 70 mL / min, 20 min) to obtain a white solid product LPi-10-6Pr-b (530 mg, yield: 58%).

[0541] LCMS(ESI): m / z, 953.4 [M] + Rt = 0.91 min.

[0542] Step 3: Preparation of compound LPi-10-6Pr

[0543] At room temperature, ethyl diisopropylamine (81.37 mg, 0.630 mmol, 2.0 eq) was added to a DMF (6.0 mL) solution of compound LPi-10-6Pr-b (300 mg, 0.315 mmol, 1.0 eq) and maleimide-PEG2-succinimide ester (223.07 mg, 0.630 mmol, 2.0 eq), and the mixture was stirred at room temperature for 10 min. The reaction solution was then directly purified by high performance liquid chromatography (JB-C235-04 column, Bonnasil-BS C18, 30 × 150 mm, 5 μm) using mobile phase A: water (0.1% TFA) and mobile phase B: acetonitrile, with acetonitrile elution ratios ranging from 19% to 40%, at 25 mL / min for 30 min, to obtain a white solid product LPi-10-6Pr (202.63 mg, yield: 54%).

[0544] LPI-10-6Pr: LCMS (ESI): m / z, 1192.2[M] + , Rt=1.099min.

[0545] 1 H NMR (400MHz, DMSO-d6) δ9.04(s,1H),8.95(s,1H),8.30(s,1H),8.17(s,1H),8.10–8.00(m,2H),7.92(t,J=5.6Hz,1H),7.31(d d,J=16.8,9.0Hz,1H),7.20(d,J=18.8Hz,1H),7.06–6.90(m,4H),5.05(s,2H),4.95(s,2H),4.77(d,J=7.6Hz,1H),3.97(d,J= 8.0Hz,5H),3.85(d,J=9.6Hz,2H),3.56(s,3H),3.55–3.51(m,4H),3.50–3.23(m,13H),3.13(s,2H),2.92(s,3H),2.72(s,2H) ,2.53(d,J=7.0Hz,1H),2.28(t,J=6.6Hz,2H),2.00-1.92(m,4H),1.13-1.03(m,2H),0.83(d,J=6.6Hz,3H),0.66-0.55(m,6H).

[0546] Example 19: Preparation of compound LPi-10-6MeiP

[0547] Step 1: Preparation of compound LPi-10-6MeiP-a

[0548] At room temperature, ethyl diisopropylamine (252.0 mg, 1.951 mmol, 5.0 eq) and HOBT (52.72 mg, 0.390 mmol, 1.0 eq) were added to a DMF (8.0 mL) solution of PI-10-6MeiP (200 mg, 0.390 mmol, 1.0 eq) and compound LP-R1-e (356.55 mg, 0.390 mmol, 1.0 eq), and the reaction was carried out at room temperature for one hour. The reaction solution was quenched with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined ethyl acetate layers were washed with saturated brine (50 mL) and dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (methanol:dichloromethane = 0–8%, 50 mL / min, 40 min) to give the yellow solid product compound LPi-10-6MeiP-a (345 mg; yield: 68.69%).

[0549] LCMS(ESI): m / z, 1287.4 [M+H] + Rt = 1.71 min.

[0550] Step 2: Preparation of compound LPi-10-6MeiP-b

[0551] To a solution of compound LPi-10-6MeiP-a (517 mg, 0.402 mmol, 1.0 eq) in acetonitrile (12.0 mL) / H2O (8.0 mL), an aqueous solution of 1N NaOH (3.2 mL, 3.2 mmol, 8.0 eq) was added, and the mixture was stirred at room temperature for one hour under ice bath conditions. The reaction solution was then purified directly by C18 reversed-phase column chromatography (mobile phase A: H2O (0.01% TFA), mobile phase B: acetonitrile, elution ratio of mobile phase A: 5–50%, 70 mL / min, 20 min) to obtain a white solid product LPi-10-6MeiP-b (320 mg, yield: 86%).

[0552] LCMS(ESI): m / z, 924.3 [M] + Rt = 1.136 min.

[0553] Step 3: Preparation of compound LPi-10-6MeiP

[0554] At room temperature, ethyl diisopropylamine (44.71 mg, 0.346 mmol, 2.0 eq) was added to a DMF (6.0 mL) solution of compound LPi-10-6MeiP-b (160 mg, 0.173 mmol, 1.0 eq) and maleimide-PEG2-succinimide ester (122.58 mg, 0.346 mmol, 2.0 eq) and stirred at room temperature for 10 minutes. The reaction solution was directly purified by high performance liquid chromatography (JB-C235-04 column, Bonnasil-BS C18, 30×150mm, 5μm, mobile phase A: water (0.1% TFA), mobile phase B: acetonitrile, elution ratio of acetonitrile from 19% to 33%, elution rate 25mL / min, 30min) to obtain a white solid product LPi-10-6MeiP (103.89mg, yield: 51%).

[0555] LPi-10-6MeiP: LCMS (ESI): m / z, 1164.4[M] + Rt = 3.767 min.

[0556] 1 H NMR (400MHz, DMSO-d6) δ9.04(s,1H),8.94(s,1H),8.27(s,1H),8.17(s,1H),8.10(d,J=9.3Hz,1H),8.00(d,J=9.4Hz,1H),7.92 (t,J=5.6Hz,1H),7.33(d,J=7.5Hz,1H),7.19(s,1H),7.02(s,1H),7.01(s,2H),5.05(s,2H),4.94(s,2H),4.78(d,J=7.5Hz,1H) ,4.02(s,3H),3.98(d,J=7.8Hz,2H),3.85(d,J=9.6Hz,2H),3.56(s,3H),3.55-3.27(m,17H),3.12(s,2H),2.92(s,3H),2.57(s, 3H), 2.53 (d, J = 7.2Hz, 1H), 2.28 (t, J = 6.5Hz, 2H), 2.02 (s, 3H), 1.98-1.88 (m, 1H), 0.81 (d, J = 6.6Hz, 3H), 0.58 (d, J = 6.8Hz, 3H).

[0557] Example 20: Preparation of compound LPI-10-6TP

[0558] Step 1: Preparation of compound LTP-b

[0559] Oxone (4206.28 mg, 6.842 mmol) was added to a solution of compound LTP-a (300 mg, 0.684 mmol) in THF (100 mL) and H₂O (100 mL) at 25 °C. The mixture was stirred overnight at 25 °C. LCMS showed that the reaction was complete. The resulting solution was extracted with DCM / MeOH = 10:1 (300 mL × 3), and the combined organic layers were washed with brine (150 mL). The organic layers were dried over Na₂SO₄, and the resulting solution was concentrated under reduced pressure to give crude compound LTP-b (100 mg, 0.170 mmol, 25%) as a yellow oil.

[0560] LCMS(ESI): m / z, 470.9 [M+H] + , Rt=1.174min.

[0561] Step 2: Preparation of compound LPI-10-6TP

[0562] Diisopropylethylamine (0.042 mL, 0.256 mmol) was added to a 2 mL DMF solution of compound LPi-10-6Et-b (80 mg, 0.085 mmol) and compound LTP-b (100.21 mg, 0.213 mmol) under ice bath conditions. The mixture was stirred at 25 °C for 10 min. LC-MS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (0–100% ACN / H₂O) to give the white solid product LPI-10-6TP (37.5 mg, yield: 31%).

[0563] LPI-10-6TP: LCMS(ESI): m / z, 1294.6[M+H] + Rt = 1.219 min.

[0564] 1H NMR (400MHz, MeOD) δ9.01(s,1H),8.61(d,J=8.6Hz,1H),8.37(d,J=8.6Hz,1H),8.20(s,1H),8.08(d,J=8.9Hz,1H),8.00(dd,J=14. 1,5.6Hz,2H),7.12(d,J=8.0Hz,1H),7.01(d,J=8.4Hz,1H),6.90(dd,J=8.3,1.6Hz,2H),5.11(s,2H),4.99(s,2H),4.62(s,1H),4. 52(d,J=4.8Hz,1H),4.18(s,2H),3.80(s,1H),3.78–3.70(m,6H),3.63(dt,J=11.2,6.1Hz,5H),3.53(dd,J=17.7,8.7Hz,2H),3.49 –3.33(m,8H),3.02(d,J=4.5Hz,3H),2.58-2.41(m,4H),2.30(s,3H),2.11(dq,J=13.5,6.8Hz,1H),1.09(s,3H),0.88-0.77(m,6H).

[0565] Example 21: Preparation of compound LPI-10-6TP2

[0566] Step 1: Preparation of compound LPI-10-6TP2

[0567] Diisopropylethylamine (0.011 mL, 0.065 mmol) was added to a DMF (1 mL) solution of compound LPi-10-6MeiP-b (30 mg, 0.032 mmol) and compound LTP-b (19.84 mg, 0.042 mmol) under ice bath conditions. The mixture was stirred at 25 °C for 18 hours. LC-MS showed that the reaction was complete. The reaction solution was filtered and purified by high-performance liquid chromatography (HPLC) (column: Ultimate C18, 21.2 × 250 mm, 5 μm; mobile phase A: water (0.1% TFA); mobile phase B: acetonitrile:B: 5-55%, 70 mL / min). The purified product was lyophilized to give a white solid product LPi-10-6TP2 (20.6 mg, yield: 47%).

[0568] LPI-10-6TP2: LCMS (ESI): m / z, 1280.6[M+H] + , Rt = 1.115 min.

[0569] 1H NMR (400MHz, CD3OD) δ9.02(s,1H),8.61(d,J=8.6Hz,1H),8.37(d,J=8.6Hz,1H),8.10(s,2H),8.04(s,1H),7.95(d, J=9.4Hz,1H),6.97(d,J=42.5Hz,4H),5.11(s,2H),4.98(s,2H),4.62(s,1H),4.32(s,1H),4.14(s,2H),3.75(t,J=5 .9Hz,3H),3.69(s,3H),3.66-3.58(m,5H),3.55-3.46(m,3H),3.44(s,3H),3.40(d,J=6.8Hz,3H),3.02(s,3H),2.88 (s,3H),2.51-2.45(m,4H),2.24(s,3H),2.01(dd,J=13.3,6.8Hz,1H),0.84(d,J=6.6Hz,3H),0.76(d,J=6.8Hz,3H).

[0570] Example 22: Preparation of compound LPI-10-6TP3

[0571] Step 1: Preparation of compound LPI-10-6TP3

[0572] Diisopropylethylamine (12.4 mg, 0.096 mmol) was added to a DMF (1 mL) solution of compound LPi-10-6-b (30.0 mg, 0.032 mmol) and compound LTP-b (15.0 mg, 0.056 mmol) under ice bath conditions. The mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was filtered and purified by high-performance liquid chromatography (HPLC) (column: Ultimate C18, 21.2 × 250 mm, 5 μm, mobile phase A: water (0.1% TFA); mobile phase B: acetonitrile:B: 13-38%, 70 mL / min). The purified product was lyophilized to give a white solid product LPI-10-6TP3 (19.2 mg, yield: 46%).

[0573] LPI-10-6TP3: LCMS (ESI): m / z, 1294.3[M+H] + Rt = 1.09 min.

[0574] 1H NMR (400MHz, CD3OD) δ9.01 (s, 1H), 8.60 (d, J = 8.4Hz, 1H), 8.36 (d, J = 8.4Hz, 1H), 8.18-8.06 (m, 8H), 8.02(s,1H),7.95(d,J=9.2Hz,1H),7.16-6.87(m,4H),5.09(s,2H),4.97(s,1H),4.66-4.62(m,1H), 4.29(d,J=2.8Hz,1H),4.20–4.07(m,2H),3.82(s,1H),3.74(t,J=5.6Hz,2H),3.68(s,3H),3.66-3.4 9(m,7H),3.48-3.35(m,8H),2.99(s,3H),2.90(s,3H),2.51-2.42(m,4H),2.27(s,3H),0.83(s,9H).

[0575] Example 23: Preparation of compound LPI-10-6SPr

[0576] Step 1: Preparation of compound LPI-10-6SPr

[0577] Diisopropylethylamine (22.1 mg, 0.171 mmol) was added to a 2 mL solution of compound LPi-10-6Pr-b (64.8 mg, 0.068 mmol) and compound LTP-b (27.0 mg, 0.057 mmol) in DMF. The mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was purified by silica gel column chromatography (C18, 0.1% TFA, H2O:ACN = 51%:49%) to give a white solid product LPi-10-6SPr (21.9 mg, yield: 29%).

[0578] LPI-10-6SPr: LCMS (ESI): m / z, 1308.0[M+H] + , Rt=1.12min.

[0579] 1H NMR (400MHz, DMSO-d6) δ9.02(s,2H),8.79(d,J=8.6Hz,1H),8.54(s,1H),8.32(s,1H),8.12(s,1H),7.96(s ,1H),7.54(s,2H),7.34(s,1H),7.13(s,2H),6.95(s,2H),5.79(s,1H),5.22(s,1H),4.97(s,2H),4.74(d,J =29.4Hz,3H),3.96-3.69(m,4H),3.58(s,5H),3.47(s,5H),3.44-3.40(m,3H),2.97(s,3H),2.82-2.52(m, 7H), 2.28 (s, 3H), 1.94 (d, J = 33.3Hz, 2H), 1.80 (s, 3H), 1.18 (s, 3H), 0.97 (s, 2H), 0.79 (s, 3H), 0.50 (s, 6H).

[0580] Example 24: Preparation of compound LPI-10-6Bu

[0581] Step 1: Preparation of compound LPi-10-6Bu-a

[0582] At 25 °C, HOBt (12.18 mg, 0.09 mmol) and ethyl diisopropylamine (0.075 mL, 0.451 mmol) were added to a DMF (1 mL) solution of compound PI-10-6Bu (78.23 mg, 0.09 mmol, 1.0 eq) and compound LP-R1-e (82.3 mg, 0.09 mmol, 1.0 eq). The mixture was stirred at 25 °C for 1 hour. LCMS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give the crude product. The residue was purified by preparative HPLC (20–70% ACN / (H2O, 0.1% TFA)) to give the white solid compound LPi-10-6Bu-a (90 mg, yield: 75%).

[0583] LCMS(ESI): m / z, 1330.3 [M+H] + Rt = 2.519 min.

[0584] Step 2: Preparation of compound LPi-10-6Bu-b

[0585] NaOH (21.66 mg, 0.542 mmol) was added to an acetonitrile (1.0 mL) / H₂O (1.0 mL) solution of compound LPi-10-6Bu-a (90 mg, 0.068 mmol) under ice bath conditions. The mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (20–70% ACN / (H₂O, 0.1% TFA)) to give a white solid compound LPi-10-6Bu-b (50 mg, yield: 76%).

[0586] LCMS (ESI): m / z, 967.6[M]+, Rt=1.111min.

[0587] Step 3: Preparation of compound LPi-10-6Bu

[0588] Diisopropylethylamine (0.021 mL, 0.124 mmol) was added to a DMF (1 mL) solution of compound LPi-10-6Bu-b (40 mg, 0.041 mmol) and compound LTP-b (23.6 mg, 0.050 mmol) at 25 °C. The mixture was stirred at 25 °C for 1 hour. LCMS showed that the reaction was complete. The resulting solution was concentrated under reduced pressure to give a crude product. The crude product was purified by preparative HPLC (JB-C235-04 column, Bonnasil-BS C18, 20 × 250 mm, 8 μm) with mobile phase A: water (0.1% FA) and mobile phase B: acetonitrile, with acetonitrile elution ratios ranging from 10% to 45% to give a white solid product LPI-10-6Bu (8.2 mg, yield: 15%).

[0589] LPI-10-6Bu: LCMS (ESI): m / z, 1322.5[M+H] + , Rt=1.981min.

[0590] 1H NMR (400MHz, CD3OD) δ8.60(d,J=8.7Hz,1H),8.35(d,J=8.4Hz,1H),8.13(s,1H),7.63(d,J=9.2Hz,3H),7.03(s,4H),5.09(s,2H),4. 89(s,2H),4.68–4.61(m,1H),4.15(d,J=8.3Hz,1H),3.94(s,2H),3.73(t,J=5.8Hz,3H),3.64–3.59(m,7H),3.58–3.51(m,2H),3.48 (dd,J=3.3,1.6Hz,2H),3.45(s,3H),3.44–3.42(m,1H),3.13(dd,J=3.3,1.7Hz,3H),2.92(s,3H),2.75(s,2H),2.55(t,J=6.6Hz,2H ), 2.46 (t, J = 5.7Hz, 2H), 2.03 (s, 1H), 1.96 (s, 3H), 1.10 (s, 4H), 0.91 (d, J = 6.6Hz, 3H), 0.73 (t, J = 6.9Hz, 3H), 0.63 (d, J = 6.6Hz, 3H).

[0591] Example 25: Preparation of compound LPI-B1T

[0592] Step 1: Preparation of compound LPi-B1T

[0593] Diisopropylethylamine (8.52 mg, 0.066 mmol) was added to a DMF (1 mL) solution of compound LP-R1-g (20 mg, 0.022 mmol) and compound LTP-b (10.3 mg, 0.022 mmol) under ice bath conditions. The mixture was stirred at 25 °C for 2 hours. LCMS showed that the reaction was complete. The reaction solution was purified by high performance liquid chromatography (flow rate: 20 mL / min, 25 °C, instrument: Gilson preparative instrument, column: Ultimate XB-C18, 21.2 × 250 mm, 5 μm (PARP-02), mobile phase A: water (0.1% TFA); mobile phase B: acetonitrile, B: 13-38%) to give a white solid LPi-B1T (6.1 mg, yield: 22%).

[0594] LPI-B1T: LCMS(ESI):m / z, 633.2[M / 2+H] + Rt = 1.08 min.

[0595] 1HNMR(400MHz,CD3 OD)δ9.02(s,1H),8.60(d,J=8.4Hz,1H),8.36(d,J=8.4Hz,1H),8.17(s,1H),8.08–7.97(m,2H) ,7.86(d,J=9.2Hz,1H),7.14–6.80(m,4H),5.11(s,3H),4.98(d,J=7.2Hz,2H),4.67–4.52(m,1 H),4.15(s,1H),3.96–3.86(m,2H),3.82–3.71(m,3H),3.66–3.52(m,9H),3.46–3.35(m,6H),3 .02(s,3H),2.82–2.75(m,1H),2.72–2.64(m,1H),2.52–2.42(m,4H),2.05(s,3H),0.74(s,9H).

[0596] Example 26: Synthesis of compound LPI-10-6

[0597] Step 1: Preparation of compound LPi-10-6-a

[0598] At room temperature, LP-R1-e (74.2 mg, 0.085 mmol), DIEA (55.2 mg, 0.427 mmol), and HOBT (11.6 mg, 0.085 mmol) were added to a 3 mL solution of PI-10-6 (45.0 mg, 0.085 mmol) in DMF. The mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was directly purified by reverse-phase column chromatography (C18) (mobile phase A: 0.1% TFA / H2O, mobile phase B: acetonitrile, B: 5-50%) to give a white solid LPi-10-6-a (48.0 mg, yield: 43%).

[0599] LCMS(ESI), m / z = 1301.6 [M+H] + Rt = 1.528 min.

[0600] Step 2: Preparation of compound LPi-10-6-b

[0601] At 0 °C, a solution of NaOH (13.5 mg, 0.338 mmol) in H₂O (2 mL) was added to a 2 mL acetonitrile solution of LPi-10⁻⁶-a (55.0 mg, 0.042 mmol). The mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was directly purified by reverse-phase column chromatography (C18, mobile phase A: 0.1% TFA / H₂O, mobile phase B: acetonitrile, B: 5-35%) to give a yellow solid LPi-10⁻⁶-b (35.0 mg, yield: 88%).

[0602] LCMS(ESI), m / z = 939.4 [M+H] + Rt = 1.052 min.

[0603] Step 3: Preparation of compound LPI-10-6

[0604] At room temperature, LP-R1-g-1 (19.8 mg, 0.056 mmol) and DIEA (14.5 mg, 0.112 mmol) were added to a DMF (3 mL) solution of LPi-10-6-b (35.0 mg, 0.037 mmol), and the mixture was stirred at 25 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was purified by reverse-phase preparation (flow rate: 20 mL / min, 25 °C, column: Ultimate XB-C18, 21.2*250 mm, 5 μm (PARP-02), mobile phase A: water (0.1% TFA); mobile phase B: acetonitrile, B: 13-38%) to give a white solid LPi-10-6 (12.3 mg, yield: 28%, TFA salt).

[0605] LCMS(ESI), m / z = 1178.0 [M+H] + Rt = 1.101 min.

[0606] 1H NMR (400MHz, CD3OD) δ8.24-8.05(m,3H),7.99(d,J=9.2Hz,1H),7.21-6.84(m,4H),6.79(s, 2H),5.16(s,2H),5.00(s,2H),4.68(s,2H),4.33(s,1H),4.18(s,2H),3.83(s,1H),3.71(s, 3H),3.67(t,J=6.0Hz,2H),3.64-3.58(m,3H),3.57-3.52(m,4H),3.51-3.42(m,8H),3.03( s,3H),2.95(s,3H),2.56(t,J=6.8Hz,2H),2.42(t,J=6.0Hz,1H),2.30(s,3H),0.85(s,9H).

[0607] ADC fabrication

[0608] Procedure 1A, trastuzumab, DAR8

[0609] Example 27 Preparation of A01030-LPi-B1

[0610] At 25°C, an aqueous solution of tris(2-carboxyethyl)phosphine (TCEP) (40 mM, 2.334 μL, 93.3 nmol) was added to a PBS buffer solution (pH = 7.4, 0.01 M) of trastuzumab (Herceptin, Genentech, product number A01030, 10.0 mg / mL, 0.2 mL, 13.3 nmol), and the solution was placed in a water bath and reacted at 25°C for 2 hours, after which the reaction was stopped.

[0611] Compound LPi-B1 (2.52 mg, 1995 nmol) was dissolved in 124.7 μL of DMSO. 12.5 μL of this solution was added to the reaction mixture, and the mixture was placed in a water bath and reacted at 25 °C for 60 minutes. The reaction mixture was purified using an ultrafiltration centrifuge tube (15 mL, 30 KD, JETBIOFIL / FTT130150) to obtain T-LPi-B1 histidine buffer (5.071 mg / mL, 0.28 mL, 20 mM Histidine-HCl, pH 5.5), which was stored at 4 °C and kept at -80 °C for long-term preservation.

[0612] The purity was determined to be 98.5% by SEC-HPLC.

[0613] Average drug loading calculated by HIC-HPLC: n = 7.8

[0614] Procedure 1B, Trastuzumab, DAR4

[0615] Preparation of Example 28A06028-LPI-10-6TP-D4

[0616] At 25°C, an aqueous solution of tris(2-carboxyethyl)phosphine (TCEP) (40 mM, 11.7 μL, 467 nmol) was added to a PBS buffer solution (pH = 7.4, 0.01 M PBS buffer solution) containing the antibody (Trastuzumab-A141C-C220S) (US9527927B2, No. A06028, 10.0 mg / mL, 1.0 mL, 66.7 nmol). The solution was placed in a water bath and reacted at 25°C for 2 hours, after which the reaction was stopped.

[0617] The compound LPI-10-6TP (0.69 mg, 533 nmol) was dissolved in 33.3 μL of DMSO and added to the above reaction solution. The mixture was placed in a water bath and reacted at 25 °C for 90 minutes. The reaction solution was purified using an ultrafiltration centrifuge tube (BIOFIL, 30K, MWCO) to obtain histidine buffer (11.48 mg / mL, 0.78 mL, 20 mM Histidine-HCl, pH 5.5) of A06028-LPI-10-6TP-D4, which was stored at 4 °C and kept at -80 °C for long-term storage.

[0618] The purity was determined to be 98.7% by SEC-HPLC.

[0619] The average drug loading calculated by HIC-HPLC: n = 3.9.

[0620] DAR value measurement

[0621] The average drug loading (DAR) was calculated by HIC-HPLC. The HIC-HPLC instrument was an Agilent 1200 or Waters ACQ-rQSM / ACQ-rFTN / W2998 high-performance liquid chromatograph; the column was a TOSOH TSKgel Bulty-NPR (2.7 μm, 4.6 × 50 mm); mobile phase A was 20 mM phosphate + 2 M ammonium sulfate buffer (pH 6.5), and mobile phase B was 20 mM phosphate buffer (pH 6.5). Gradient elution was performed with mobile phase B-isopropanol (80:20) (0-2 min 10% B / 10% C, 2-15 min 10% B / 10% C-75% B / 25% C, 15-18 min 75% B / 25% C), the flow rate was 0.6 mL / min, the detection wavelength was 280 nm, and the column temperature was 25 °C.

[0622] Example 29

[0623] The ADC A01030-LPi-7-1 shown in Table 1 was obtained by referring to the experimental method of Example 27 above.

[0624] Example 30

[0625] The ADCs A06028-LPI-10-6TP2-D4, A06028-LPI-10-6TP3-D4, A06028-LPI-10-6SPr-D4, A06028-LPI-10-6Bu-D4, A06028-LPI-10-6-D4, A06028-LPI-B1T-D4, and A06028-LPI-B1-D4 shown in Table 1 were obtained by referring to the experimental method of Example 28 above.

[0626] Table 1 List of relevant ADCs

[0627] Test Example 1: Inhibitory activity of the compound of the present invention against cancer cells

[0628] (1) Experimental steps

[0629] Day 0: Cells in good growth condition were digested, sieved, and counted. 2000 or 1000 cells / 100μL of culture medium were placed in each well of a 96-well plate (N87 gastric cancer cells, Shanghai Academy of Sciences Type Culture Collection, SCSP-534; JIMT-1 breast cancer cells, Nanjing Kebai Biotechnology Co., Ltd., CBP60378; MDA-MB-468 cells, Shanghai Academy of Sciences Type Culture Collection, TcHu136). 200μL of sterile water or PBS was added to the outer ring, and the cells were incubated for 24h.

[0630] Day 1: The test molecules were serially diluted to 2 μM with culture medium, followed by a fourfold serial dilution in 9 wells. The 10th well served as a negative control for culture medium. 100 μL of the diluted sample was taken out with a pipette and gently piped into the incubated Day 0 cell plate along the edge of the well. The plate was then placed in an incubator for incubation.

[0631] Day 7: 144 hours after cell treatment, the chromogenic reagent CellCounting-Lite 2.0 (brand: Vazyme, catalog number: DD1101-03) was dissolved 30 minutes in advance. 100 μL was added to each well and the cells were developed in the dark for 2-5 minutes. The readings were taken twice and the average was calculated. The results are shown in Table 2.

[0632] (2) Experimental Results

[0633] The results showed that the small molecule NMT inhibitors of this application have good anticancer activity, especially PI-7-1, PI-R1dH, PI-7-1iP-P2, PI-10-6iP, PI-10-6MeiP, PI-10-6Et, PI-10-6Pr, PI-10-6Bu, and PI-10-6, which have excellent anticancer activity.

[0634] Table 2 shows the IC50 values ​​of the compounds in this application against N87 gastric cancer cells, MDA-MB-468 and JIMT-1 breast cancer cells. 50 value

[0635] Test Example 2: Efficacy Experiment of the ADC of the Present Invention in the JIMT-1 Xenograft Model of Human Breast Cancer

[0636] Human breast cancer JIMT-1 cell line (Nanjing Kebai Biotechnology, CBP60378) was cultured in DMEM medium (Gibco, catalog number: 11995-065) + 10% FBS (Excell Bio, catalog number: FCS500) at 37°C in an incubator containing 5% CO2. Cells were digested with 0.25% trypsin-EDTA, washed twice with PBS, counted using a cell counter, and diluted with PBS to a cell concentration of 1×10⁻⁶. 8 / mL.

[0637] CB17 SCID mice, female, 4-6 weeks old, were purchased from Vital River Laboratory Animal Co., Ltd. After acclimatization to the laboratory environment for 2-5 days, CB17 SCID mice were subcutaneously inoculated with JIMT-1 cells at a dose of 5 × 10⁻⁶ cells per rib. 6 / animal, inoculated with 0.1 mL (containing 50% Matrigel), until the tumor grows to 150-200 mm 3 When the medication is administered according to the regimen shown in Table 3, the DAR value is adjusted to 4 for all doses.

[0638] Table 3. Dosing regimens of ADCs in human breast cancer JIMT-1 xenograft mice.

[0639] At the end of the experiment, mice were euthanized, and tumors were collected and weighed. Tumor volume and body weight were measured twice a week, and tumor proliferation rate and relative tumor inhibition rate were calculated. The calculation formula is as follows:

[0640] Relative tumor proliferation rate T / C (%) = T RTV / C RTV ×100%

[0641] (T RTV The average RTV in the treatment group; C RTV The average RTV of the control group; RTV = V t / Vo V o V represents the tumor volume of the animal when grouped. t (This refers to the tumor volume in the animal after treatment);

[0642] Relative tumor inhibition rate (TGI%) = (1 - T / C) × 100%

[0643] (T and C represent the relative tumor volume (RTV) of the treatment group and the control group at a specific time point, respectively).

[0644] The formula for calculating tumor volume (V) is: V = l / 2 × L 长 ×L 短 2 ,

[0645] Where L 长 L 短 These represent the long diameter and short diameter of the tumor, respectively.

[0646] The experimental results are shown in Table 4 and Figure 1.

[0647] Table 4. Tumor volume data of the JIMT-1 breast cancer-bearing mouse model.

[0648] The results showed that the ADCs of this application have good anticancer activity, especially A06028-LPI-10-6TP-D4 and A06028-LPI-10-6TP2-D4, which have excellent anticancer activity.

[0649] Test Example 3: Evaluation of Membrane Permeability of the Compounds of the Invention

[0650] Experimental methods:

[0651] 1. Preparation of the drug solution

[0652] The 10.0 mM stock solution was diluted with DMSO to obtain a 0.200 mM working solution.

[0653] 760 μL of PBS was added to 40 μL of working solution to obtain a 10.0 μM dosing solution. The final concentration of DMSO was 5%.

[0654] 2. Add 150 μL of 10.0 μM drug delivery solution to each well of the donor plate, which is pre-coated with 5 μL of 1% porcine brain polar lipid extract / dodecane mixture on a polyvinylidene fluoride membrane.

[0655] 3. Add 300 μL of PBS to each well of the PTFE receptor plate.

[0656] 4. After the donor plate and recipient plate are combined, they are incubated at room temperature with shaking at 300 rpm for 4 hours.

[0657] 5. Preparation of T0 sample: Transfer 20 μL of donor solution to a new well, and then add 230 μL of PBS and 130 μL of stop solution (containing acetonitrile internal standard) to prepare T0 sample.

[0658] 6. Receptor sample preparation: Remove the plate from the incubator. Transfer 250 μL of solution from each receptor well and mix with 130 μL of stop solution to prepare the receptor sample.

[0659] 7. Preparation of donor samples: Transfer 20 μL of solution from each donor well and mix it with 230 μL of PBS and 130 μL of stop solution as the donor sample.

[0660] 8. All samples were analyzed by LC-MS / MS.

[0661] Data Analysis:

[0662] The equation used to determine permeability (Pe) is as follows: [drug] acceptor =(A a / A i ×DF)acceptor; [drug] donor =(A a / A i *DF)donor;

[0663] A a / A i : Analyze the area ratio of the internal standard in the analysis text; DF: dilution factor. [drug]equilibrium=([drug] donor ×VD+[drug] acceptor ×VA) / (VD+VA)

[0664] Where V D V is the volume of the donor well (0.150 mL); A The volume of the acceptor pore (0.300 mL); the area is the effective surface area of ​​the membrane (0.300 cm²). 2 ); time refers to the incubation time (14400 seconds in this experiment); C A and C D These are the area ratios of the control compound or test compound in the recipient compartment and the donor compartment, respectively; C0 is the initial area ratio of the control compound or test compound in the donor compartment.

[0665] Experimental results: The permeability of the compounds of this invention in the BBB PAMPA model is shown in Table 5.

[0666] Table 5 Summary of Permeability Evaluation of Compounds of the Invention Note: CNS-:P e <15.0nm / s; CNS+:P e ≥15.0nm / s

[0667] Experimental conclusions: The compounds PI-10-6MeiP, PI-10-6Et, PI-10-6Pr and PI-10-6 of this invention exhibit good permeability in the BBB PAMPA model, especially PI-10-6MeiP and PI-10-6Et, which have superior permeability compared to PI-R1.

[0668] Test Example 4: In vitro liver microsomal stability test of the compound of the present invention

[0669] Experimental methods:

[0670] Transfer the microparticle solution (100 μL) to all reaction plates (blank, T0, T5, T15, T30, T45, T60, and NCF60).

[0671] Add 2 μL of the compound working solution to all 96-well reaction plates except for the blanks (T0, T5, T15, T30, T45, T60 and NCF60).

[0672] All reaction plates containing the mixture of compounds and microparticles were pre-incubated at 37°C for 10 minutes.

[0673] Add 98 μL of 100 mM potassium phosphate buffer to the NCF60 reaction plate.

[0674] The NCF60 reaction plate was incubated at 37°C for 60 minutes.

[0675] After pre-incubation, add 98 μL of cofactor working solution to each reaction plate except NCF60 (blank, T0, T5, T15, T30, T45 and T60).

[0676] The reaction plate was incubated at 37°C.

[0677] Add stop solution (600 μL) at the appropriate end time point for each reaction plate to terminate the reaction.

[0678] Shake all the sample plates for 10 minutes, then centrifuge at 3220×g for 20 minutes at 4°C.

[0679] The supernatant from each reaction plate was transferred to eight new 96-well plates for LC-MS / MS analysis.

[0680] Data Analysis:

[0681] The remaining percentage of the compound after incubation is calculated using the following formula:

[0682] T is calculated using the first-order dynamic equation. 1 / 2 C t =C0·e -k·t

[0683] Experimental results: The stability test results of the liver microsomes of the compound of this invention are shown in Figure 6.

[0684] Table 6 Summary of the liver microsomal stability of the compounds of the present invention

[0685] Experimental conclusions: The compounds of this invention exhibit faster clearance rates in liver microsomes of various genera. After exerting their anti-tumor proliferation effects in tumor tissue, the toxins of ADC drugs can be rapidly cleared from the liver, reducing toxicity. In particular, PI-10-6MeiP, PI-10-6Et, PI-10-6Pr, and PI-10-6 are cleared more rapidly than PI-R1.

Claims

1. An NMT inhibitor, which is a compound of formula (I), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt thereof. in: R1 is a group of formula -XLA; L represents -(CR9R) 10 )r-3-8-membered cycloalkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-8-membered cycloalkylene-O-*、-(CR9R 10 )r-3-8-membered heterocyclic alkylene-O-*、-(CR9R 10 )tO-* or -(CR9R 10 )t-(NR 11 )s-*; * represents the linking site with group A, r is 0, 1, 2, 3 or 4, s is 0 or 1, t is 1, 2, 3, 4, 5 or 6; each R9, R 10 Each of these can be independently H, halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, -COOH, or -CON(R). 11 2. 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; R 11 H, optionally substituted C1-8 alkyl, -COOH, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl; The cycloalkylene, heteroalkylene, cycloalkylene, and heteroalkylene groups may optionally be substituted with substituents selected from the following: halogen, amino, hydroxyl, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino. X represents -O-, -NR 11 -, -S- or a single key; A is a 6-10 aryl or a 5-10 heteroaryl group, wherein the aryl or heteroaryl group is optionally substituted with a substituent selected from the following: halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino, optionally substituted C1-8 alkylacyl, -COOH, -CON(R) 11 )2; Y is either C or N; Each R2 is independently a halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio or optionally substituted C1-8 alkylamino; m is 0, 1, 2 or 3 or 4. K, Z, and M are each independently C or N; Each R3 is independently a halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, or optionally substituted C1-8 alkylamino; n is 0, 1, 2, or 3. Each R4 is independently H or an optional substituted C1-8 alkyl group, and p is 1, 2 or 3; Each R5 and R6 is independently H, halogen, hydroxyl, amino, nitro, cyano, mercapto, or optionally substituted C1-8 alkyl, and q is 1, 2, 3, or 4. R7 and R8 are each independently H or optionally substituted C1-8 alkyl groups; The optional substituents in the C1-8 alkyl, C1-8 alkoxy, C1-8 alkylthio, C1-8 alkylamino, and C1-8 alkylacyl groups are selected from the following groups: halogen, hydroxyl, amino, nitro, cyano, mercapto, C1-8 alkoxy, C1-8 alkylamino, C1-8 alkylacyl, -COOH, -CON(R) 11 2, 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl.

2. The NMT inhibitor according to claim 1, characterized in that, L represents -(CR9R) 10 )r-3-6-membered cycloalkylene-(NR 11 )s-*、-(CR9R 10 )r-3-6-membered heterocyclic alkylene-(NR) 11 )s-*、-(CR9R 10 )r-3-6-membered cycloalkylene-O-*、-(CR9R 10 )r-3-6-membered heterocyclic alkylene-O-* or -(CR9R 10 )t-(NR 11 )s-*; r is 0, 1 or 2, s is 0 or 1, t is 1, 2 or 3.

3. The NMT inhibitor according to claim 1 or 2, characterized in that, Each R9, R 10 Each of these can be independently H, C1-8 alkyl, hydroxyl-substituted C1-8 alkyl, amino-substituted C1-8 alkyl, C1-8 alkoxy-substituted C1-8 alkyl, C1-8 alkylamino-substituted C1-8 alkyl, -COOH-substituted C1-8 alkyl, 3-6 membered cycloalkyl, or 3-6 membered heterocycloalkyl; R 11 It can be H, C1-8 alkyl, hydroxyl-substituted C1-8 alkyl, amino-substituted C1-8 alkyl, C1-8 alkoxy-substituted C1-8 alkyl, C1-8 alkylamino-substituted C1-8 alkyl, -COOH-substituted C1-8 alkyl, 3-6 membered cycloalkyl or 3-6 membered heterocycloalkyl.

4. The NMT inhibitor according to claim 3, characterized in that, L represents a 3-6 membered cycloalkylene group, -(CR9R) 10 )r-3-6-membered cycloalkylene*, -3-6-membered cycloalkylene-NR 11 -* or -(CR9R) 10 )r-3-6-membered cycloalkylene-NR 11 -*; r is 1 or 2.

5. The NMT inhibitor according to claim 3, characterized in that, L represents -(CR9R) 10 )t-NR 11 -* or -(CR9R) 10 )t-, where t is 1, 2, or 3, optionally, R9, R 10 R 11 At least one of them is a hydroxyl-substituted C1-8 alkyl, an amino-substituted C1-8 alkyl, a C1-8 alkoxy-substituted C1-8 alkyl, a C1-8 alkylamino-substituted C1-8 alkyl, a -COOH-substituted C1-8 alkyl, a 3-6 membered cycloalkyl, or a 3-6 membered heterocycloalkyl; preferably, R9, R 10 R 11 Each can be independently H, methyl, ethyl, n-propyl, n-butyl, hydroxymethyl, or hydroxyethyl.

6. The NMT inhibitor according to claim 3, characterized in that, L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H, C1-4 alkyl; preferably, L represents -(CH2)2-NR 11 *, R 11 H represents methyl, ethyl, n-propyl, or n-butyl.

7. The NMT inhibitor according to claim 1, characterized in that, L represents -CH2CH2-NH-*, -CH2CH2-N(-CH3)-*, -CH(-CH2OH)-CH2-N(-CH3)-*, -CH2CH2-N(-CH2CH3)-*, -CH2CH2-N(-CH2CH2CH3)-, -CH2CH2-N(-CH2CH2CH2CH3)-*, -CH2-CH(-CH2OH)-N(-CH3)-*, -CH2CH2-N(-CH2CH2OH)-*, -CH2CH2-* or -CH2CH2-O-*; * represents the linking site with group A.

8. The NMT inhibitor according to any one of claims 1-7, characterized in that, A is a 6-10 aryl or a 5-10 heteroaryl, wherein the aryl or heteroaryl group is optionally substituted with a substituent selected from the following: C1-8 alkyl, C1-8 alkylacyl, -COOH, -CON(R) 11 )2, and those affected by hydroxyl, C1-8 alkyl acyl, -COOH, -CON(R) 11 2. C1-8 alkyl groups substituted with 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl.

9. The NMT inhibitor according to any one of claims 1-8, characterized in that, A is a 5-6-membered heteroaryl group, wherein the heteroaryl group contains 1, 2 or 3 N atoms; preferably, A is a 4-pyrazolyl group.

10. The NMT inhibitor according to claim 9, characterized in that, A is Among them, R 12 R 13 Each is independently selected from H, halogen, hydroxyl, amino, nitro, cyano, mercapto, optionally substituted C1-8 alkyl, optionally substituted C1-8 alkoxy, optionally substituted C1-8 alkylthio, optionally substituted C1-8 alkylamino or optionally substituted C1-8 alkylacyl. R 14 R 15 R 16 R 17 Each is independently selected from H, halogen, hydroxyl, amino, nitro, cyano, mercapto, or optionally substituted C1-8 alkyl groups; v is 0, 1, or 2; The substituents in the optionally substituted C1-8 alkyl, C1-8 alkoxy, C1-8 alkylthio, C1-8 alkylamino, and C1-8 alkylacyl groups are selected from the following groups: halogen, hydroxyl, amino, nitro, cyano, and mercapto.

11. The NMT inhibitor according to claim 10, characterized in that, R 12 R 13 Each is independently selected from H, C1-8 alkyl; R 14 R 15 R 16 Each is independently selected from H, C1-8 alkyl; R 17 Selected from C1-8 alkyl groups; v is 0, 1 or 2.

12. The NMT inhibitor according to claim 11, characterized in that, R 12 R 13 Each is independently selected from C1-8 alkyl groups; R 16 Selected from H;R 17 Selected from C1-8 alkyl groups; v is 0; preferably, R 12 R 13 Each is independently selected from methyl, R 17 Selected from isopropyl or tert-butyl.

13. The NMT inhibitor according to claim 9, characterized in that, A is R 12 R 13 Each is independently selected from C1-4 alkyl groups; R 16 Selected from H;R 17 Selected from C1-4 alkyl groups; v is 0; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups; Preferably, R 12 R 13 Each is independently selected from methyl, R 17 Selected from isopropyl or tert-butyl, L represents -(CH2)2-NR 11 *, R 11 It can be H, methyl, ethyl, n-propyl, or n-butyl; More preferably, R 12 R 13 Each is independently selected from methyl, R 17 Selected from isopropyl or tert-butyl, L represents -(CH2)2-NR 11 *, R 11 It can be methyl or ethyl.

14. The NMT inhibitor according to claims 1-13, characterized in that, Each R2 is independently a halogen; m is 0, 1, 2; preferably, each R2 is independently F or Cl; m is 2.

15. The NMT inhibitor according to claims 1-14, characterized in that, K and Z are N, and M is C; or K, Z, and M are all N; or K is C, and Z and M are N.

16. The NMT inhibitor according to claims 1-15, characterized in that, The compound of formula (I) is either a compound of formula (I-1) or formula (I-2):

17. The NMT inhibitor according to claim 16, characterized in that, The compound of formula (I) is either a compound of formula (I-3) or a compound of formula (I-4): Preferably, each R2 is independently F or Cl, R 16 Selected from H, R 17 It is selected from isopropyl or tert-butyl, and R7 and R8 are each independently H or C1-8 alkyl.

18. The NMT inhibitor according to claim 17, characterized in that, Each R2 is independently F, R7 and R8 are H or methyl, respectively; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups; Preferably, the compound of formula (I) is a compound of formula (I-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be H, methyl, ethyl, n-propyl, or n-butyl; More preferably, the compound of formula (I) is a compound of formula (I-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be methyl or ethyl.

19. The NMT inhibitor according to claim 1, characterized in that, Compound (I) is one of the following:

20. An NMT inhibitor, which is a compound of formula (II), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt thereof. in: Q represents a linker group that can be conjugated to antibody or antigen binding fragments; L1 represents the linking group; The definitions of other groups are as defined in any one of claims 1-19.

21. The NMT inhibitor according to claim 20, characterized in that, Q represents a linker group containing a maleimide structure, a linker group containing a methanesulfonylpyrimidine structure, a linker group containing a methanesulfonylpyridinothiazole structure, a linker group containing a methanesulfonylpyrimidineothiazole structure, a linker group containing a cycloenyne structure, or a linker group containing an oxime bond structure. Preferably, Q-L1- represents 22. The NMT inhibitor according to claim 20 or 21, characterized in that, The compound of formula (II) is either a compound of formula (II-1) or a compound of formula (II-2).

23. The NMT inhibitor according to claim 22, characterized in that, The compound of formula (II) is either a compound of formula (II-3) or a compound of formula (II-4). Preferably, each R2 is independently F or Cl, R 16 Selected from H, R 17 It is selected from isopropyl or tert-butyl, and R7 and R8 are each independently H or C1-8 alkyl.

24. The NMT inhibitor according to claim 23, characterized in that, Each R2 is independently F, R7 and R8 are H or methyl, respectively; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups; Preferably, the compound of formula (II) is a compound of formula (II-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be H, methyl, ethyl, n-propyl, or n-butyl; More preferably, the compound of formula (II) is a compound of formula (II-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be methyl or ethyl.

25. The NMT inhibitor according to claim 20, characterized in that, Compounds of formula (II) are the following compounds:

26. An antibody-drug conjugate, characterized in that, It is formed by conjugating the NMT inhibitor described in any one of claims 20-25 with an antibody or antigen-binding fragment Ab via a linker group.

27. The antibody-drug conjugate according to claim 26, characterized in that, It is a compound of formula (III), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer, or mixture of isomers thereof, or a pharmaceutically acceptable salt thereof. in: Ab represents an antibody or antigen-binding fragment; L' is a direct bond or a linking group connecting Ab and Q'; Q' is the group formed by connecting Q to Ab through L'; w is an integer or decimal between 0 and 20; preferably an integer or decimal between 0 and 8. Other groups are defined as described in any one of claims 20-25.

28. The antibody-drug conjugate according to claim 27, characterized in that, Ab is selected from murine antibodies, chimeric antibodies, humanized antibodies, fully human antibodies, or their antigen-binding fragments.

29. The antibody-drug conjugate according to claim 28, characterized in that, The antibody (Ab) is selected from monoclonal antibodies and polyclonal antibodies; preferably, the antibody is selected from anti-HER2 (ErbB2) antibody, anti-EGFR antibody, anti-B7-H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-ROR1 antibody, anti-CLDN6 antibody, anti-CLDN9 antibody, anti-CLDN18.2 antibody, anti-NaPi-2b antibody, anti-TNF-α antibody, anti-ENPP3 antibody, anti-DLL3 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD28 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD38 antibody, anti-CD44 antibody, anti-CD45 antibody, anti-CD47 antibody, anti-CD48 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD98 antibody, anti-CD105 antibody, anti-CEA antibody, anti-EphA2 antibody, anti-MUCI antibody, and anti-Lewis antibody. Anti-γ antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-CD79 antibody, anti-TROP-2 antibody, anti-CD79B antibody, anti-Mesothelin antibody, anti-Nectin-4 antibody, anti-TPBG antibody, or their antigen-binding fragments.

30. The antibody-drug conjugate according to claim 29, characterized in that, Ab is selected from monoclonal antibodies or their antigen-binding fragments, preferably from trastuzumab, cetuximab, pertuzumab, nimotuzumab, enoblituzumab, emibetuzumab, Inotuzumab, pinatuzumab, brentuximab, gemtuzumab, bivatuzumab, morovatuzumab, or their antigen-binding fragments.

31. The antibody-drug conjugate according to any one of claims 26-30, characterized in that, The compound of formula (III) is either a compound of formula (III-1) or a compound of formula (III-2). Preferably, the compound of formula (III) is a compound of formula (III-3) or a compound of formula (III-4). More preferably, each R2 is independently F or Cl, R 16 Selected from H, R 17 It is selected from isopropyl or tert-butyl, and R7 and R8 are each independently H or C1-8 alkyl.

32. The antibody-drug conjugate according to claim 31, characterized in that, Each R2 is independently F, R7 and R8 are H or methyl, respectively; L represents -(CH2)t-NR 11 -*, t is 1, 2, or 3, R 11 Selected from H and C1-4 alkyl groups; Preferably, the compound of formula (III) is a compound of formula (III-3), where L represents -(CH2)2-NR. 11 *, R 11 H represents methyl, ethyl, n-propyl, or n-butyl. More preferably, the compound of formula (III) is a compound of formula (III-3), where L represents -(CH2)2-NR. 11 *, R 11 It can be methyl or ethyl.

33. The antibody-drug conjugate according to claim 27, characterized in that, The compound of formula (III) is one of the following compounds:

34. A pharmaceutical composition comprising an NMT inhibitor according to any one of claims 1-25, or an antibody-drug conjugate according to any one of claims 26-33, and one or more pharmaceutically acceptable carriers or excipients.

35. Use of the NMT inhibitor according to any one of claims 1-25 in the preparation of antibody-drug conjugates.

36. Use of the NMT inhibitor according to any one of claims 1-25, or the antibody-drug conjugate according to any one of claims 26-33, or the pharmaceutical composition according to claim 34 in the preparation of a medicament for the prevention or treatment of tumors or cancer.

37. A method for preventing or treating tumors or cancer, comprising administering to a subject in need an effective amount of an NMT inhibitor according to any one of claims 1-25, an antibody-drug conjugate according to any one of claims 26-33, or a pharmaceutical composition according to claim 34.

38. The use according to claim 36, or the method according to claim 37, characterized in that, The tumor or cancer is selected from solid tumors or hematologic malignancies; preferably selected from breast cancer, gastric cancer, colon cancer, pancreatic cancer, kidney cancer, ovarian cancer, lung cancer, soft tissue sarcoma, liposarcoma, melanoma, head and neck cancer, cervical cancer, and prostate cancer; more preferably selected from HER-2 positive breast cancer, triple-negative breast cancer, HER-2 positive triple-negative breast cancer, renal clear glandular adenocarcinoma, small cell lung cancer, non-small cell lung cancer, or large cell lung cancer.

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