WO2025190395A2

WO2025190395A2 - Mta-cooperative prmt5 inhibitor

Info

Publication number: WO2025190395A2
Application number: PCT/CN2025/082618
Authority: WO
Inventors: Cheng-hao LIU; Long-zhi LIN; Hong-ren WANG; Jianfeng MOU; Xinli DUAN; Qingfang YU; Tong Wang; Bin Chen; Peiyu Zhang
Original assignee: PharmaEngine Inc
Current assignee: PharmaEngine Inc
Priority date: 2024-03-14
Filing date: 2025-03-14
Publication date: 2025-09-18
Anticipated expiration: 2026-09-14
Also published as: WO2025190395A3

Abstract

The present disclosure relates to compounds used as MTA-cooperative inhibitors of protein arginine N-methyl transferase 5 (PRMT5). In particular, the present invention relates to MTA-cooperative PRMT5 inhibitors, including pharmaceutical compositions comprising the compounds and the methods of use thereof.

Description

MTA-COOPERATIVE PRMT5 INHIBITOR

TECHNICAL FIELD

The present invention relates to compounds used as MTA-cooperative inhibitors of protein arginine N-methyl transferase 5 (PRMT5) . In particular, the present invention relates to MTA-cooperative PRMT5 inhibitors, including their pharmaceutical compositions comprising the compounds and the methods of use thereof.

BACKGROUND

The protein arginine methyltransferase PRMT5 is crucial for maintaining cellular homeostasis by regulating gene transcription, ribosomal biogenesis, mRNA splicing, protein translation, DNA damage response, and immune functions (Musiani, D. et al., Proteomics profiling of arginine methylation defines PRMT5 substrate specificity. Sci. Signaling 2019, 12 (575) , eaat8388) . PRMT5 is a type II arginine methyltransferase that symmetrically dimethylate histones and non-histone proteins. It catalyzes a methyl group transfer from S-adenosyl-L-methionine (SAM) to an omega-nitrogen of the guanidino function of protein L-arginine residues (omega-monomethylation) and a second methyl group to the other omega-nitrogen, resulting in symmetric dimethylarginine (sDMA) (Gary, J.D., and Clarke, S. Prog. Nucleic Acid Res. Mol. Biol 1998, RNA and protein interactions modulated by protein arginine methylation. 61, 65–131) .

Due to multiple functions involved with PRMT5, complete inactivation of PRMT5 is lethal in most cell lines. Therefore, PRMT5 is considered as an essential gene for cell viability (McDonald, E.R., et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 2017, 170 (3) , 577-592) . PRMT5 is an attractive drug target in the field of oncology. Several potent and selective inhibitors targeting the catalytic pocket of PRMT5 have been developed. However, given the essential role of PRMT5 in normal tissue homeostasis, solely inhibiting PRMT5 may cause the toxicity and lead to a limited therapeutic window.

Methylthioadenosine (MTA) is an endogenous competitor of SAM and partially inhibits PRMT5 without inhibiting PRMT family members other than PRMT5. Studies have shown that inhibition of PRMT5 by MTA will reduce the methylation activity and sensitize cells to PRMT5 inhibitors. MTA is a substrate of methylthioadenosine phosphorylase (MTAP) and can be converted into 5-methylthioribose 1-phosphate (MTR-1-P) by MTAP. Therefore, MTA will not be accumulated in the wild type cells to reduce PRMT5 activity. On the contrary, cells with homozygous deletion of MTAP will cause MTA to be accumulated in the cells, resulting in decreased PRMT5 activity (Kryukov, G.V. et al., MTAP deletion confers enhanced dependency on the arginine methyltransferase PRMT5 in human cancer cells. Science 2016, 351 (6278) , 1214-1218) . Therefore, MTA-cooperated inhibition of PRMT5 will selectively inhibit PRMT5 activity in cells with homozygous deletion of MTAP.

Homozygous deletions of MTAP are relatively common in cancers with about 15%incidence rate. Higher incidence rates were noticed in glioblastoma, bladder cancer, non-small cell lung cancer, and pancreatic cancer. This loss is due to its proximity to the P16/CDKN2A tumor suppressor locus. Homozygous deletions of p16/CDKN2a commonly involve the co-deletion of adjacent genes, such as MTAP (Mavrakis, K.J. et al., Disordered methionine metabolism in MTAP/CDKN2A-deleted cancers leads to dependence on PRMT5. Science 2016, 351 (6278) , 1208-1213) .

We realized that MTA-cooperative inhibition of PRMT5 activity in MTAP-deficient cancers will provide therapeutic benefit for a wide range of cancers. Thus, there is a need to develop new MTA-cooperative PRMT5 inhibitors that are capable of inhibiting PRMT5 activity in the MTAP-deficient cells.

SUMMARY OF THE INVENTION

The compounds of the present invention provide this therapeutic benefit as MTA-cooperative inhibitors of PRMT5 that negatively modulate the activity of MTA-bound PRMT5 in a cell, particularly an MTAP-deficient cell, or for treating various forms of MTAP-associated cancer.

One aspect of the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, L, X₁, X₂, X₃, R_A, R_B, R_C, m, n, p and q are as defined herein.

One aspect of the present invention provides a compound of formula (I-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, L, R_A, R_B, R_C, m, n, p and q are as defined herein.

One aspect of the present invention provides a compound of formula (IA-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined herein, and ring B and ring C are connected in the form of a spirocyclic structure.

One aspect of the present invention provides a compound of formula (IA-1-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, R_A, R_B, R_C, n, p and q are as defined herein, and ring B and ring C are connected in the form of a spirocyclic structure.

One aspect of the present invention provides a compound of formula (IA-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, X₁, X₂, X₃, R_A, R_B, R_C, m₁, m₂, Z₁, Z₂, Z₃, Z₄, Z₅, n, p1 and q are as defined herein. In one aspect, ring B and ring C of formula (I) form a spiro ring to give formula (IA-1-2) .

One aspect of the present invention provides a compound of formula (IB-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined herein.

One aspect of the present invention provides a compound of formula (IB-1-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, R_A, R_B, R_C, n, p and q are as defined herein.

One aspect of the present invention provides a compound of formula (IB-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p1 and q are as defined herein.

One aspect of the present invention provides a compound of formula (IC-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined herein; ring B and ring C are fused to form a fused tricyclic structure.

One aspect of the present invention provides a compound of formula (IC-1-1) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, ring B, ring C, R_A, R_B, R_C, n, p and q are as defined herein; ring B and ring C are fused to form a fused tricyclic structure.

One aspect of the present invention provides a compound of formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein ring A, X₁, X₂, X₃, W₁, W₂, W₃, W₄, W₅, W₆, W₇, m₃, m₄, R_A, R_B, R_C, n, p and q are as defined herein.

Another aspect of the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, and one or more pharmaceutically acceptable carriers or excipients. The pharmaceutical composition of the present invention is used as an MTA-cooperative PRMT5 inhibitor.

Another aspect of the invention provides the use of a compound of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, in the manufacture of a drug or a medicament of MTA-cooperative PRMT5 inhibitors

Another aspect of the invention provides a method for treating cancer, the method comprising administering a therapeutically effective amount of a compound of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled to a subject in need thereof to inhibit the growth of the cancer in the subject. The compounds of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) , a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled may be used alone or in combination with another therapeutic agent and/or therapy.

Another aspect of the invention provides use of a compound of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, in the manufacture of a medicament for treating cancer. In one aspect, the medicament further comprises another therapeutic agent, or the medicament is used in combination with another therapeutic agent and/or therapy.

Another aspect of the invention provides a compound of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, for treating cancer

Another aspect of the invention provides a pharmaceutical composition for treating cancer, the pharmaceutical composition comprising formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled.

Another aspect of the invention provides a method for preparing a compound of formula (I) , formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) , or formula (IC-1-2) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled.

DETAILED DESCRIPTION OF THE INVENTION

The present invention can be understood easily by referring to the following detailed description of various embodiments of the invention, the examples, and the tables with their relevant descriptions. Unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms such as those defined in commonly used dictionaries should be interpreted consistently with their meaning in the context of the relevant art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

Definition

The definitions set forth in this section are intended to clarify terms used throughout this application. The term "herein" means the entire application.

It must be noted that, as used herein, the singular forms "a, " "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, unless otherwise required by context, singular terms shall include the plural and plural terms shall include the singular.

Usually, ranges are expressed herein as from "about" one particular value and/or to "about" another particular value. When such a range is expressed, an embodiment includes the range from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the word "about, " it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to and independently of the other endpoint. As used herein, the term "about" refers to ± 20%, preferably ± 10%, and even more preferably ± 5%.

As used herein, the phrase "optionally substituted" means that substitution is optional. In the event a substitution is desired, such substitution means that any number of hydrogens on the designated atom are replaced with a selection from the indicated group, provided that the normal valence of the designated atom is not exceeded, and that the substitution results in a stable compound. For example, when a substituent is keto (i.e., =O) , 2 hydrogens on the atom are replaced. Examples of substituents for a "substituted" group are those found in the exemplary compounds and embodiments disclosed herein and can include, but are not limited to, halogen, cyano, alkyl, alkoxy, haloalkyl, alkylamino, aminoalkyl, dialkylamino, hydroxyalkyl, alkoxyalkyl, hydroxyalkoxy, alkoxyalkoxy, aminoalkoxy, alkylaminoalkoxy, alkylaminoalkyl and the like.

As used herein, the term "halogen" includes fluorine, chlorine, bromine and iodine. "Halo, " used as a prefix of a group, means one or more hydrogens on the group are replaced with one or more halogens.

The term "alkyl" used herein refers to a monovalent, saturated, straight or branched hydrocarbon group containing 1 to 12 carbon atoms. Preferably, the alkyl is a C₁-C₈ alkyl group. More preferably, the alkyl is a C₁-C₆ alkyl group. The alkyl can be unsubstituted or substituted with one or more substituents. Examples of a C₁-C₆ alkyl group include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl (including all isomeric forms) , and hexyl (including all isomeric forms) , heptyl (including all isomeric forms) , octyl (including all isomeric forms) and the like.

Unless the number of rings is specified otherwise, the term "cycloalkyl" used herein refers to a saturated, monovalent hydrocarbon group having cyclic configurations, including monocyclic, bicyclic, tricyclic, and higher polycyclic alkyl groups (and, when multicyclic, including fused and bridged bicyclic and spirocyclic moieties) wherein each cyclic moiety has from 3 to 12 carbon atoms. Preferably, the cycloalkyl has from 3 to 8 carbon atoms. More preferably, the cycloalkyl has from 3 to 6 carbon atoms. When cycloalkyl contains more than one ring, the rings may be fused or unfused and include bicyclic groups. Fused rings generally refer to at least two rings sharing two atoms therebetween. Such cycloalkyl groups include, for example, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl and the like, or bicyclic, polycyclic or bridged ring structures, such as adamantyl and the like.

Unless the number of rings is otherwise specified, the term "aryl" means a monovalent 6-14 membered mono-or bicyclic ring, such as an 8-11 membered mono-or bicyclic ring, wherein at least one of the rings in the bicyclic ring is a carbon aromatic ring. Representative examples include, but are not limited to, phenyl, biphenyl, naphthyl, indenyl, benzocyclopentyl and the like.

Unless the number of rings is otherwise specified, "heteroaryl" means a monocyclic, fused bicyclic or fused tricyclic monovalent group of 5 to 14 ring atoms, which contains one or more, for example one, two, three or four, ring heteroatom moieties independently selected from the following: -O-, -S (O) _n- (n is 0, 1 or 2) , -N-, -N (R^x) -, and the remaining ring atoms are carbon, wherein the ring containing the monocyclic group is an aromatic ring and at least one of the fused rings containing the bicyclic or tricyclic group is an aromatic ring. One or two ring carbon atoms of any non-aromatic ring containing the bicyclic or tricyclic group may be replaced by -C (O) -, -C (S) -or -C (=NH) -groups. R^x is hydrogen, C₁-C₈ alkyl, hydroxy, C₁-C₈ alkoxy, C₁-C₈ acyl or C₁-C₈ alkylsulfonyl. More preferably, C₁-C₈ alkyl is C₁-C₆ alkyl, C₁-C₈ alkoxy is C₁-C₆ alkoxy, C₁-C₈ acyl is C₁-C₆ acyl, C₁-C₈ alkylsulfonyl is C₁-C₆ alkylsulfonyl. Fused bicyclic groups include bridged ring systems. Unless otherwise indicated, valences may be located on any atom of any ring of the heteroaryl group as long as valence rules permit. Specifically, when the valence position is located on nitrogen, R^x is absent. More specifically, the term "heteroaryl" includes, but is not limited to, 2, 3-dihydrobenzofuranyl, 2, 3-dihydrofuro [3, 2-b] pyridine, 2, 3-dihydrofuro [3, 2-c] pyridine, 6, 7-dihydro-5H-cyclopenta [b] pyridine, chromanyl, isochromanyl, 3, 4-dihydro-2H-pyrano [2, 3-b] pyridine, 5, 8-dihydro-6H-pyrano [3, 4-b] pyridine, 1, 3, 4, 5-tetrahydrobenzo [c] oxazepine, phthalidyl, indolinyl, 1, 2, 4-triazolyl, 1, 3, 5-triazolyl, phthalimide, pyridyl, pyrrolyl, imidazolyl, thienyl, furanyl, indolyl, 2, 3-dihydro-1H-indolyls (including, for example, 2, 3-dihydro-1H-indol-2-yl or 2, 3-dihydro-1H-indol-5-yl, etc. ) , isoindolyl, dihydroindolyl, isoindolyl, benzimidazolyl, benzodioxol-4-yl, benzofuranyl, cinnolinyl, indolizinyl, naphthyridin-3-yl, phthalazin-3-yl, phthalazin-4-yl, pteridinyl, purinyl, quinazolinyl, quinoxalinyl, tetrazolyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, diazinyl, oxazolyl, isoxazolyl, oxadiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyls (including, for example, tetrahydroisoquinolin-4-yl or tetrahydroisoquinolin-6-yl, etc. ) , pyrrolo [3, 2-c] pyridinyls (including, for example, pyrrolo [3, 2-c] pyridin-2-yl or pyrrolo [3, 2-c] pyridin-7-yl, etc. ) , benzopyranyl, thiazolyl, isothiazolyl, thiadiazolyl, benzothiazolyl, benzothiophenyl, tetrahydrofuropyridyl, pyridotetrahydrofuranyl, tetrahydropyranopyridyl, pyridotetrahydropyranyl, tetrahydropyranophenyl or benzotetrahydropyranyl, pyridoxazolyl, oxazolopyridyl, pyridopyrrol, pyrrolopyridyl, oxadiazolphenyl, benzodiazolyl, benzooxazolyl, oxazolophenyl, and derivatives thereof, or N-oxides or protected derivatives thereof.

Unless the number of rings is otherwise specified, the term "heterocyclyl" means a saturated or partially unsaturated monovalent monocyclic group of 3 to 9 ring atoms or a saturated or partially unsaturated monovalent fused bicyclic group of 5 to 12 ring atoms, wherein one or more, such as one, two, three or four, ring heteroatoms are independently selected from the following: -O-, -S (O) _x- (x is 0, 1 or 2) , -N=, -N (R^y) - (wherein R^y is hydrogen, C₁-C₈ alkyl, hydroxy, C₁-C₈ alkoxy, C₁-C₈ acyl or C₁-C₈ alkylsulfonyl) , more preferably, C₁-C₈ alkyl is C₁-C₆ alkyl, C₁-C₈ alkoxy is C₁-C₆ alkoxy, C₁-C₈ acyl is C₁-C₆ acyl, C₁-C₈ alkylsulfonyl is C₁-C₆ alkylsulfonyl, and the remaining ring atoms are carbon. One or two ring carbon atoms may be replaced by -C (O) -, -C (S) -, or -C (=NH) -groups. Fused bicyclic groups include bridged ring systems. Unless otherwise stated, the valence of a group may be located on any atom of any ring within the group as long as the valence rules allow. Specifically, when the valence position is located on the nitrogen atom, Ry is absent. More specifically, the term "heterocyclyl" includes, but is not limited to, piperidinyl, pyrimidinyl, morpholinyl, piperazinyl, azetidinyl, pyrrolidinyl, 2-oxopyrrolidinyl, 2, 5-dihydro-1H-pyrrolyl, 4-piperidonyl, 2-oxopiperazinyl, dihydropyranyl, tetrahydropyranyl, 2-oxopiperidinyl, thiomorpholinyl, thiomorpholinyl, perhydroazepinyl, pyrazolidinyl, dihydroimidazolyl, imidazolidinyl, dihydropyridinyl, tetrahydropyridinyl, oxazolinyl, oxazolidinyl, isoxazolidinyl, thiazolinyl, thiazolidinyl, quinuclidine, isothiazolidinyl, octahydroindolyl, octahydroisoindolyl, decahydroisoquinolinyl, tetrahydrofuranyl and tetrahydropyranyl, and derivatives thereof, as well as N-oxides or protected derivatives thereof. The term "ring" as used herein refers to "cycloalkyl" , "aryl" , "heteroaryl" , "heterocyclic group" , and "cycloalkyl ring" , "aromatic ring" , "heteroaromatic ring" , "heterocyclic ring" which do not exist in the form of a radical.

As used herein, the term "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, wherein the parent compound is modified by preparing a pharmaceutically acceptable acid or base salt thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali metal or organic salts of acidic residues such as carboxylic acids; and the like. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Suitable nontoxic acids include, but are not limited to, inorganic and organic acids such as acetic acid, alginic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, formic acid, fumaric acid, furoic acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, propionic acid, salicylic acid, stearic acid, succinic acid, p-aminobenzenesulfonic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid. Non-limiting examples of salts of the compounds of the invention include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, 2-hydroxyethanesulfonate, phosphate, hydrogenphosphate, acetate, adipate, alginate, aspartate, benzoate, butyrate, camphorate, camphorsulfonate, citrate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, succinate, malonate, fumarate, maleate, methanesulfonate, mesitylenesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, trimethylacetate, propionate, trichloroacetate, trifluoroacetate, glutamate, bicarbonate, undecanoate, lactate, citrate, tartrate, gluconate, benzenesulfonate, and p-toluenesulfonate.

As used herein, the term "geometric isomers" includes, but is not limited to, cis and trans; E-and Z-forms; c-, t-, and r-forms; internal and external; R-, S-and meso; boat, chair, twist, envelope and semi-chair; and combinations thereof.

As used herein, the term "enantiomers" refers to a pair of stereoisomers that are nonsuperimposable mirror images of each other. A 1: 1 mixture of a pair of enantiomers is a racemic mixture. The term "enantiomers" is used where appropriate to refer to racemic mixtures. A "diastereomer" is a stereoisomer having at least two atoms that are asymmetric but are not mirror images of each other. Absolute stereochemistry can be assigned according to the Cahn-Ingold-Prelog R-Ssystem. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be designated by R or S. Resolved compounds can be represented as (+) or (-) depending on the direction (dextral or levorotary) which they rotate plane polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers or axes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R) -or (S) -. The present invention is meant to include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R) -and (S) -isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be in the E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis-or trans-configuration.

As used herein, "prodrugs" are intended to include any covalently bonded carriers that release the active parent drug according to the compounds described herein through in vivo physiological action, such as hydrolysis, metabolism and the like, when such prodrug is administered to a subject. The applicability and technology involved in preparing and using prodrugs are well known to those of ordinary skill in the art. Prodrugs of the compounds of formula (I) (parent compounds) can be prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compounds. "Prodrugs" include the compounds described herein wherein a hydroxy, amino, or sulfhydryl group is bonded to any group that, when the prodrugs are administered to a subject, cleaves to form a free hydroxyl, free amino, or free sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, derivatives and metabolites of the compounds described herein that include biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogues. In certain embodiments, prodrugs of the compounds of described herein with carboxyl functional groups are the lower alkyl (e.g., C₁-C₆) esters of the carboxylic acid. The carboxylate esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule.

The present invention also includes all pharmaceutically acceptable isotopically labeled compounds identical to those of the present invention except that one or more atoms are replaced by atoms with the same atomic number but different atomic masses or mass numbers from those that predominate in nature. Examples of isotopes suitable for inclusion in the compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium (D, ²H) , tritium (T, ³H) ) ; isotopes of carbon (e.g., ¹¹C, ¹³C, and ¹⁴C) ; isotopes of chlorine (e.g., ³⁶Cl) ; isotopes of fluorine (e.g., ¹⁸F) ; isotopes of iodine (e.g., ¹²³I and ¹²⁵I) ; isotopes of nitrogen (e.g., ¹³N and ¹⁵N) ; isotopes of oxygen (e.g., ¹⁵O, ¹⁷O and ¹⁸O) ; isotopes of phosphorus (e.g., ³²P) ; and isotopes of sulfur (e.g., ³⁵S) . Certain isotopically labeled compounds of the present invention (e.g., those incorporating radioactive isotopes) may be used in drug and/or substrate tissue distribution studies (e.g., assays) . The radioactive isotopes tritium (i.e., ³H) and carbon-14 (i.e., ¹⁴C) are particularly useful for this purpose because they are easy to incorporate and detect. Substitution with positron-emitting isotopes such as ¹¹C, ¹⁸F, ¹⁵O, and ¹³N can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the invention may be prepared by methods analogous to those described in the accompanying Schemes and/or Examples and Preparations by using appropriate isotopically labeled reagents in place of the previously employed non-labeled reagents. Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted, for example, D₂O, acetone-d₆ or DMSO-d6.

As used herein, the term "solvate" means a compound or a pharmaceutically acceptable salt thereof that further includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. If the solvent is water, the solvate may be suitably called a "hydrate, " such as hemi-hydrate, mono-hydrate, sesqui-hydrate, di-hydrate, tri-hydrate, etc.

Compounds

In one embodiment, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein:

X₁, X₂ and X₃ are each independently selected from CR and N; preferably X₁, X₂ and X₃ are each independently selected from CR;

ring A, which is fused withis selected from a 5-6 membered cycloalkyl, a 5-6 membered heterocyclyl, a 5-6 membered aryl and a 5-6 membered heteroaryl, or is absent, and preferably ring A is selected from a 5-membered heterocyclyl and a 5-6 membered heteroaryl or is absent, and more preferably ring A is 1, 2-oxadiazole, 1, 2-thiazole, tetrahydrofuran or pyridine; whereindenotes the position fused with ring A;

ring B is a 5-6 membered heterocyclic group, and preferably the heteroatom of ring B is connected to L;

ring C is selected from a 5-6 membered heterocyclyl, a 5-6 membered aryl, a 5-6 membered heteroaryl, a 8-11 membered fused bicyclic heterocyclyl, a 8-11 membered fused bicyclic aryl and a 8-11 membered fused bicyclic heteroaryl;

L is -C (=O) -;

denotes the connection between ring B and ring C; the connection is selected from: (1) a direct bond; (2) ring B and ring C being fused to form a fused tricyclic ring;

and (3) ring B and ring C forming a spiro ring;

each R is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably hydrogen, deuterium, halogen or C₁-C₆ alkyl;

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, oxo (=O) , cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably hydrogen, deuterium, amino or C₁-C₆ alkyl;

each R_B is independently hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, amino, C₁-C₆ alkyl, C₃-C₆ cycloalkyl-C₁-C₆ alkyl-, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , or

two R_B attached to the same carbon atom together with the carbon atom to which they are attached form a C₃-C₈ cycloalkyl group, or

R_B attached to different carbon atoms together with the carbon atoms to which they are attached form a C₃-C₇ cycloalkyl group;

each R_C is independently hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, mercapto, amino, C₁-C₆ alkyl optionally substituted with 1-3 R_Z, methylene optionally substituted with 1-3 R_Z, C₂-C₆ alkenyl optionally substituted with 1-3 R_Z, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ alkylthio, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -NH- (C₁-C₆ haloalkyl) , -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , C₁-C₆ haloalkoxy, C₁-C₆ haloalkylthio, pentafluorosulfur, -S (O) ₂- (C₁-C₆ alkyl) , -S (=O) (C₁-C₆ alkyl) (=N) R_Z, -P (=O) (C₁-C₆ alkyl) ₂, C₂-C₆ haloalkenyl, 3-6 membered cycloalkyl optionally substituted by 1-3 R_Z, 5-8 membered bridged cycloalkyl optionally substituted by 1-3 R_Z, 4-7 membered heterocyclyl optionally substituted by 1-3 R_Z, 5-6 membered aryl optionally substituted by 1-3 R_Z, 5-6 membered heteroaryl optionally substituted by 1-3 R_Z, or

two R_C connected to the same carbon atom together with the carbon atom to which they are connected form a C₃-C₈ cycloalkyl, or

R_C connected to different carbon atoms together with the carbon atom to which they are connected form a C₃-C₇ cycloalkyl or a 3-7 membered heterocyclyl, or

two R_C connected to the same carbon atom together with the carbon atom to which they are connected form C=T, wherein T is selected from CR_ER_F, and R_E and R_F are each independently selected from hydrogen, deuterium, halogen and C₁-C₆ alkyl;

each R_Z is independently selected from hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, amino, pentafluorosulfur, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , 3-6 membered cycloalkyl, -NH- (C₁-C₆ haloalkyl) and -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) ;

m is 0 or 1; wherein when m is 0, it means that L is a direct bond; when m is 1, L is -C (=O) -, and ring B and ring C are fused to form a fused tricyclic ring;

n is 0, 1, 2, 3 or 4;

p is 0, 1, 2, 3 or 4; and

q is 0, 1, 2, 3 or 4.

In the preceding embodiment, the compound has formula (I-1) ,

wherein ring A, ring B, ring C, L, m, R_A, R_B, R_C, n, p and q are as defined above; preferably:

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl , - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl.

In any preceding embodiments, the compound has formula (IA-1) ,

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; and ring B and ring C form a spiro ring.

In any preceding embodiments, is the compound has formula (IA-1-1) ,

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; and ring B and ring C form a spiro ring; preferably:

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl.

In any preceding embodiments, ring B is a 5-6 membered heterocyclic group containing at least one N; preferably:

is selected from more preferably

wherein *denotes the spiral atom; and

R_B and p are as defined above.

In any preceding embodiments, p is at least 1, and at least one R_B is an oxo group; preferably, ring B is a 5-6 membered heterocyclic group containing at least one N, p is at least 1, and at least one R_B is an oxo group; more preferably:

is selected from

wherein *denotes the spiral atom.

In any preceding embodiment, ring C is selected from a fused bicyclic ring formed by a 5-membered ring fused to a 6-membered ting and a fused bicyclic ring formed by a 6-membered ring fused to a 6-membered ring; preferably, ring C is selected from a fused bicyclic ring formed by: a 5-membered heterocyclyl fused to a 6-membered aryl, a 5-membered heterocyclyl fused to 6-membered heteroaryl, a 6-membered heterocyclyl fused to 6-membered aryl, and a 6-membered heterocyclyl fused to 6-membered heteroaryl; more preferably is selected from

wherein *denotes the spiral atom;

q1 is 0, 1, 2 or 3; and

R_C and q are as defined above.

In any preceding embodiments,

is selected from

wherein *represents the spiral atom.

In any preceding embodiments, the compound has formula (IA-1-2) ,

wherein:

Z₁ is selected from CH₂, NH and O, wherein H is optionally replaced by R_C;

Z₂, Z₃, Z₄ and Z₅ are each independently selected from CH and N, wherein H is optionally replaced by R_C;

m₁ and m₂ are each independently 0 or 1, preferably when one of m₁ and m₂ is 0, then the other is 1;

p1 is 0, 1, 2 or 3; and

ring A, X₁, X₂, X₃, R_A, R_B, R_C, n and q are as defined above.

In any preceding embodiments, the compound has formula (IA-1-2-1) , formula (IA-1-2-2) , formula (IA-1-2-3) , formula (IA-1-2-4) or formula (IA-1-2-5) :

preferably

wherein ring A, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above;

p1 and q1 are independently 0, 1, 2 or 3; and

Z₂, Z₃, Z₄ and Z₅ are as defined above.

In any preceding embodiments, the compound has formula (IB-1) ,

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above.

In any embodiments, the compound has formula (IB-1-1) ,

wherein ring A, ring B, ring C, R_A, R_B, R_C, n, p and q are as defined above, preferably

is selected from

more preferably, #denotes the connection with (as defined above) , $ denotes the connection with ring C; and

R_B and p are as defined above.

is selected from

wherein #denotes the connection with (as defined above) , and $ denotes the connection with ring C.

In any preceding embodiments,

is selected from

R_c and q are as defined above.

In any preceding embodiments,

is selected from

In any embodiment, the compound has formula (IB-1-2) ,

Whereindenotes a single bond or a double bond;

p1 is 0, 1, 2 or 3; and

ring A, X₁, X₂, X₃, R_A, R_B, R_C, n and q are as defined above.

In any preceding embodiments, the compound has formula (IC-1) ,

wherein ring A, ring B, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; ring B and ring C are fused to form a fused tricyclic ring; and

ring C is selected from 8-11 membered fused bicyclic ring, preferably 8-11 membered fused bicyclic heterocyclic group, 8-11 membered fused bicyclic aryl group or 8-11 membered fused bicyclic heteroaryl group.

In any preceding embodiments, the compound has formula (IC-1-1) ,

wherein ring A, ring B, R_A, R_B, R_C, n, p and q are as defined above; preferably

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl;

ring B and ring C are fused to form a fused tricyclic ring; and

In any preceding embodiments, ring B is a 5-6 membered heterocyclic group containing at least one N; preferably: ring B and ring C are fused to form a fused tricyclic ring, and

is selected from

whereindenotes the chemical bond shared by ring B and ring C; when there is an N atom in the shared chemical bond, the shared chemical bond denoted in ring C is predominant over the shared bond denoted in ring C;

p2 is 0, 1 or 2; and

R_B and p are as defined above.

In any preceding embodiments, ring B and ring C are fused to form a fused tricyclic ring, and

is selected from

whereindenotes the chemical bond shared by ring B and ring C; when there is an N atom in the shared chemical bond, the shared chemical bond denoted in ring C is predominant over the shared bond denoted in ring C.

In any preceding embodiments, ring C is selected from a fused bicyclic ring formed by a 5-membered ring fused to a 6-membered ring and a fused bicyclic ring formed by a 6-membered ring fused to a 6-membered ring; preferably, ring C is selected from a fused bicyclic ring formed by: a 5-membered heterocyclyl fused to a 6-membered aryl, a 5-membered heterocyclyl fused to a 6-membered heteroaryl, a 6-membered heterocyclyl fused to a 6-membered aryl, a 6-membered heterocyclyl fused to a 6-membered heteroaryl, a 5-membered cycloalkyl fused to a 6-membered aryl, a 5-membered cycloalkyl fused to a 6-membered heteroaryl, a 6-membered cycloalkyl fused to a 6-membered aryl, and a 6-membered cycloalkyl fused to a 6-membered heteroaryl; more preferably, ring B and ring C are fused to form a fused tricyclic ring, and

is selected from

whereindenotes the chemical bond shared by ring B and ring C;

q1 is 0, 1, 2 or 3; and

R_C and q are as defined above.

In any preceding embodiments, ring B and ring C are fused to form a fused tricyclic ring, andis selected from

whereindenotes the chemical bond shared by ring B and ring C.

In any preceding embodiments, the compound has formula (IC-1-2) ,

wherein:

W₁ is selected from CH₂, NH and O;

W₂ is selected from C, CH and N;

W₃ is selected from CH₂, C=O, NH and O; preferably CH₂, C=O and O;

W₄, W₅, W₆ and W₇ are each independently selected from CH and N; preferably W₄, W₅, W₆, and W₇ are CH;

m₃ is 0 or 1;

m₄ is 0 or 1, preferably 0;

ring A, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; and

denotes a single bond or a double bond.

In any preceding embodiments, the compound has formula (IC-1-2-1) , formula (IC-1-2-2) , formula (IC-1-2-3) or formula (IC-1-2-4) :

preferably

more preferably

preferably

more preferably

preferably more preferably

preferablymore preferably

W₁, W₂, W₃, W₄, W₅, W₆ and W₇ are as defined above;

q1 is 0, 1, 2 or 3; and

p2 is 0, 1 or 2.

In any preceeding embodiments, wherein

is selected from

R_A and n are as defined above.

In any preceeding embodiments, wherein

is selected from

n₁ is 0, 1, 2 or 3; and

R_A, X₁, X₂, X₃, R_A, and n are as defined above;

preferably, is selected from

In any preceding embodiments, the present invention provides a compound selected from:

or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled.

In any preceding embodiments, the compound has formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) or formula (IC-1-2) .

Pharmaceutical compositions, uses and methods

The compounds or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled of the present invention can be therapeutically administered as the neat chemical, but it may be more beneficial to administer the compounds or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled in a form of a pharmaceutical composition or formulation. Thus, the present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, and one or more pharmaceutically acceptable carriers or excipients.

The pharmaceutical compositions can be administered in a variety of dosage forms including, but not being limited to, a solid dosage form, a liquid dosage form, an oral dosage form, a parenteral dosage form, an intranasal dosage form, suppository, lozenge, sugar-coated dosage form, oral administered, a controlled release dosage form, a pulsed release dosage form, an immediate release dosage form, an intravenous solution, suspension or combinations thereof. The compounds can be administered, for example, via oral or parenteral routes, including such as intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, airway (aerosol) , rectal, vaginal and topical (including buccal and sublingual) administration.

In one embodiment of the present invention, the compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled is orally administered. For oral administration, the compounds may generally be provided in unit dosage form suitable for ingestion by a subject, such as tablets, pills, sugar-coated tablets, lozenges, capsules, powders, granules, aqueous solutions, suspensions, liquids, gels, syrups, slurries and the like. The dosage form may be a tablet or a controlled release dosage form formulated as a tablet. Tablets for oral use may include the active ingredients mixed with one or more pharmaceutically acceptable excipients.

An "excipient" generally refers to a substance, often an inert substance, added to a pharmacological composition or otherwise used as a vehicle to further facilitate administration of a compound. Examples of excipients include, but are not limited to, inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents, preservatives, effervescent mixtures, and adsorbents. Suitable inert diluents include, but are not limited to, sodium carbonate, calcium carbonate, sodium phosphate, calcium phosphate, lactose, etc. Suitable disintegrating agents include, but are not limited to: starch (such as corn starch) , cross-linked polyvinylpyrrolidone, agar, alginic acid or its salts (such as sodium alginate) , etc. Binding agents may include, but are not limited to: magnesium aluminum silicate, starch (such as corn, wheat, or rice starch) , gelatin, methylcellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone, etc. If present, lubricating agents are usually magnesium stearate, calcium stearate, stearic acid, talc, or hydrogenated vegetable oil. If necessary, the tablet may be coated with a material of such as mono-stearin glycerides or di-stearin glycerides to delay absorption thereof in the gastrointestinal tract. The compositions can also be further formulated into chewable tablets.

Pharmaceutical compositions of this invention for oral use can be obtained by combining a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled with a solid excipient, optionally grinding the mixture, and processing the mixture after adding suitable additional compounds (if necessary) to give tablets or sugar-coated drug cores. Suitable solid excipients, in addition to those previously mentioned, include carbohydrates or protein fillers, including, but not being limited to, sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; celluloses such as methyl cellulose, hydroxypropylmethyl-cellulose or sodium carboxymethylcellulose; and gums including gum arabic and gum tragacanth; as well as proteins such as gelatin and collagen.

Capsules for oral administration include, but are not limited to, hard gelatin capsules comprising an active ingridient (s) mixed with a solid diluent (s) and soft gelatin capsules comprising an active ingredient (s) mixed with water or an oild (such as peanut oil, mineral oild or olive oil) .

Suitable sugar coating (s) can be applied on tablet cores. Concentrated sugar solutions, optionally further comprising gum arabic, talc, polyvinylpyrrolidone, carbopolgel, poly (ethylene glycol) and/or titanium dioxide, lacquer solution and suitable organic solvents or solvent mixtures, cab be used for this purpose. Pigments or colorants can be introduced to tablets or sugar-coatings cores to distinguish or characterize different (combination) of dosages of an active compound (s) .

The pharamceutical composition can further comprise a suitable solid or gel phase carrier. Examples of such carriers include, but are not limited to, calcium carbonate, calcium phosphate, various types of sugars, starch, cellulose derivatives, gelatin and polymers such as poly (ethylene glycol) .

Compounds and pharmaceutical compositions or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled in the present invention include an active ingredient that is administered in a therapeutically effective amount to achieve its intended purpose. The term "therapeutically effective amount" refers to that amount of a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, alone or in combination with ionizing radiation or an anticancer agent which, upon single or multiple dose administration to the subject, provides the desired effect in the subject under treatment. Toxicity and therapeutic efficacy of such compound can be determined by standard pharmacological procedures with cell cultures or experimental animals, e.g., for determining the IC₅₀ values. As used herein, "IC₅₀" refers to the concentration of an agent which can produce 50%of the maximal inhibitory response by that agent.

The actual amount of a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled to be administered will be determined by a physician under relevant circumstances, including the condition to be treated, the size and type of neoplasia or tumor, the possible routes of administration, the specific compound of the invention to be administered, timing of administration of hedgehog pathway modulators relative to other therapies, the subject's type, species, age, weight, gender and medical condition, the subject's renal and liver function, and the severity of the subject's symptoms. Achieving optimal precision in producing drug concentrations within the therapeutic range requires a scheme based on the kinetics of the drug's availability to the target site. This involves the distribution, balance, and elimination of the drug. In some cases, dosage levels below the lowest end of the foregoing ranges may be more than the sufficient amount, while in other cases higher dosages may still be employed.

A "subject" to be treated by the method of the present invention means either a human or non-human animal, such as primate, mammal, and vertebrate.

"In vivo" means within a living subject, such as within an animal or human. In this context, an agent can be used therapeutically in vivo to retard or eliminate the proliferation of aberrantly replicating cells. The agents also can be used in vivo as a prophylactic to prevent aberrant cell proliferation or the manifestation of symptoms associated therewith.

"In vitro" means outside a living subject. Examples of in vitro cell populations include cell cultures and biological samples, such as fluid or tissue samples from humans or animals. Such samples can be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine and saliva. Exemplary tissue samples include tumors and biopsies thereof. In this context, the present compounds can be employed in numerous applications, both therapeutic and experimental.

In addition, the present invention relates to a method for treating cancer in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound as described herein, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, prodrug or solvate thereof, or an isotopically labeled version of any of the foregoing.

The present invention also relates to compounds as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled for use in treating cancer.

The present invention also relates to use of compounds as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled in the manufacture of a medicament for treating cancer.

"Cancer" refers to a cell proliferative disease state that includes, but is not limited to: cardiac cancers such as sarcomas (e.g., angiosarcoma, fibrosarcoma, rhabdomyosarcoma, and liposarcoma) , myxomas, rhabdomyomas, fibromas, lipomas, and teratomas; lung cancers, such as bronchial carcinomas (e.g., squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, and adenocarcinoma) , alveolar carcinomas (e.g., bronchiolar carcinoma) , bronchial adenomas, sarcomas, lymphomas, cartilaginous hamartomas, and mesothelioma; gastrointestinal cancers, such as esophageal cancer (e.g., squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, and lymphoma) , gastric cancer (e.g., lymphoma and leiomyosarcoma) , pancreatic cancer (e.g., ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid and vasoactive intestinal peptide tumor) , small bowel cancer (e.g., adenocarcinoma, lymphoma, carcinoid, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, and fibroma) , colorectal cancer (e.g., adenocarcinoma, ductal adenoma, villous adenoma, hamartomas, and leiomyomas) ; genitourinary tract cancers, such as kidney cancer (e.g., adenocarcinoma, Wilm's tumor [nephroblastoma] , lymphoma, and leukemia) , bladder and urethra cancer (e.g., squamous cell carcinoma, transitional cell carcinoma, and adenocarcinoma) , prostate cancer (e.g., adenocarcinoma and sarcoma) , testicular cancer (e.g., sperm cell carcinoma, teratoma, embryonal tumor, teratoma, choriocarcinoma carcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, and lipoma) ; liver cancer, such as hepatocellular carcinoma (e.g., hepatocellular carcinoma) , cholangiocarcinoma, hepatoblastoma, angiosarcoma, liver cancer Cellular adenomas and hemangioma; bone cancers such as osteogenic sarcoma (e.g., osteosarcoma) , fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (e.g., reticular cellular sarcoma) , multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (e.g., osteochondral exostosis) , benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and Giant cell tumors; cancers of the nervous system, such as skull cancers (e.g., osteomas, hemangiomas, granulomas, xanthomas, and osteitis malformations) , meningeal cancers (e.g., meningiomas, meningiosarcomas, and polygliosis) , brain cancer (e.g., astrocytoma, medulloblastoma, glioma, ependymoma, blastoma (pineal tumor) , glioblastoma multiforme, oligodendroglioma, schwannomas, retinoblastomas, and congenital tumors) , spinal neurofibromas, meningiomas, gliomas, and sarcomas; gynecological cancers, such as uterine cancer (e.g., endometrial cancer) , cervical cancer (e.g., cervical cancer and preneoplastic cervical dysplasia) , ovarian cancer (e.g., serous cystadenocarcinoma, mucinous cystadenocarcinoma, and carcinoma of unknown category] , theca granulosa cell tumors, Stelli-Rediger cell tumors (Sertoli-Leydig cell tumor, dysgerminoma, and malignant teratoma) , vulvar cancer (e.g., squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, and melanoma) , vaginal cancer (e.g., clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma [e.g., embryonal rhabdomyosarcoma] , and fallopian tube cancer) ; blood cancers, such as blood cancers (myeloid leukemia [acute and chronic] , acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferation diseases, multiple myeloma and myelodysplasia) , Hodgkin's disease, non-Hodgkin's lymphoma [malignant lymphoma] ; skin cancers such as malignant melanoma, basal cell carcinoma, squamous cell carcinoma, cell carcinoma, Kaposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; adrenal adenocarcinoma, such as neuroblastoma; or breast cancer.

The present invention also relates to the use of compounds as described hereinor a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled in the manufacture of medicaments for modulating the hedgehog pathway.

The compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled can be administered as the sole active agent, or in combination with other known cancer therapy.

The term "in combination with" means that the compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled may be administered before, after, concurrently with any one or more other antineoplastic therapies, or before, after, or concurrently with any combination of other antineoplastic therapies. Thus, the compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled and the second anticancer agent may be administered as a single composition or as two separate compositions simultaneously or as two separate compositions sequentially. Likewise, a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled and chemotherapy or ionizing radiation therapy may be administered simultaneously, separately, or sequentially. One skilled in the art will understand that the amount of a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled administered with an anti-cancer therapy is preferably an amount sufficient to enhance the effects of the anti-cancer therapy or an amount sufficient to induce apoptosis or cell death in conjunction with anti-cancer therapy to maintain an anti-angiogenic effect.

The term "second anticancer agent" as used herein, unless otherwise indicated, refers to an agent (s) capable of inhibiting or preventing the growth of neoplasms or tumors, or inhibiting the maturation and proliferation of malignant (cancer) cells. Second anticancer agents suitable for use in combination with the compounds of formula (I) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled include, but are not limited to targeted cancer drugs, such as trastuzumab, ramucirumab, vismodegib, sonidegib, bevacizumab, everolimus, tamoxifen, toremifene, fulvestrant, anastrozole, exemestane, lapatinib, letrozole, pertuzumab, ado-trastuzumab emtansine, palbociclib, cetuximab, panitumumab, ziv-aflibercept, regorafenib, lmatinib mesylate, lanreotide acetate, sunitinib, regorafenib, denosumab, alitretinoin, sorafenib, pazopanib, temsirolimus, everolimus, tretinoin, dasatinib, nilotinib, bosutinib, rituximab, alemtuzumab, ofatumumab, obinutuxumab, ibrutinib, idelalisib, blinatumomab, soragenib, crizotinib, erlotinib, gefitinib, afatinib dimaleate, ceritnib, ramucirumab, nivolumab, pembrolizumab, osimertinib, and necitumumab; an alkylating agent, such as busulfan, chlorambucil, cyclophosphamide, iphosphamide, melphalan, nitrogen mustard, streptozocin, thiotepa, uracil nitrogen mustard, triethylenemelamine, temozolomide, and 2-chloroethyl-3-sarcosinamide-1-nitrosourea (SarCNU) ; an antibiotic or plant alkaloid, such as actinomycin-D, bleomycin, cryptophycins, daunorubicin, doxorubicin, idarubicin, irinotecan, L-asparaginase, mitomycin-C, mitramycin, navelbine, paclitaxel, docetaxel, topotecan, vinblastine, vincristine, teniposide (VM-26) , and etoposide (VP-16) ; a hormone or steroid, such as 5α-reductase inhibitor, aminoglutethimide, anastrozole, bicalutamide, chlorotrianisene, diethylstilbestrol (DES) , dromostanolone, estramustine, ethinyl estradiol, flutamide, fluoxymesterone, goserelin, hydroxyprogesterone, letrozole, leuprolide, medroxyprogesterone acetate, megestrol acetate, methyl prednisolone, methyltestosterone, mitotane, nilutamide, prednisolone, arzoxifene (SERM-3) , tamoxifen, testolactone, testosterone, triamicnolone, and zoladex; a synthetic, such as all-trans retinoic acid, carmustine (BCNU) , carboplatin (CBDCA) , lomustine (CCNU) , cis-diaminedichloroplatinum (cisplatin) , dacarbazine, gliadel, hexamethylmelamine, hydroxyurea, levamisole, mitoxantrone, o, p'-dichlorodiphenyldichloroethane (o, p'-DDD) (also known as lysodren or mitotane) , oxaliplatin, porfimer sodium, procarbazine, and imatinib mesylate an antimetabolite, such as chlorodeoxyadenosine, cytosine arabinoside, 2'-deoxycoformycin, fludarabine phosphate, 5-fluorouracil (5-FU) , 5-fluoro-2'-deoxyuridine (5-FUdR) , gemcitabine, camptothecin, 6-mercaptopurine, methotrexate, and thioguanine; and a biologic, such as alpha interferon, BCG (Bacillus Calmette-Guerin) , granulocyte colony stimulating factor (G-CSF) , granulocyte-macrophage colony-stimulating factor (GM-CSF) , interleukin-2, and herceptin.

Unless otherwise indicated, the term "treating" as used herein means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. Unless otherwise indicated, the term "treatment" as used herein refers to the behavior or action of "treating" as defined above.

Synthesis of Compound

The invention also relates to a process for the preparation of compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled. The compounds of the present invention can be prepared by those skilled in the art using conventional organic synthesis methods and commercially available materials.

In one embodiment, the invention relates to a method for preparing a compound as described herein or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled. Examples of the methods of preparation comprise, but are not limited to, those procedures illustrated in the examples described herein.

It is to be understood that the examples used herein are for the purpose of explanation but not intended to be limiting. Those skilled person in this field may vary the process or steps of the following examples to obtain the desired products.

Intermediate Example 1: Synthesis of intermediate 1226-6

Step 1

To a solution of compound 1226-1 (8.0 g, 29.7 mmol) in THF (80 mL) was added n-BuLi (38 mL, 60.8 mmol, 1.6 M) dropwise at -78 ℃ under N₂. After the reaction solution was stirred at -78 ℃ for 5 hours, N, N-dimethyl formamide (DMF, 3.0 mL, 38.6 mmol) was added. After stirring for additional 3 hours, the reaction solution was poured slowly into cold water (80 mL) . The resulting aqueous solution was washed with EtOAc, acidified to pH = 3 using 1 M aq. HCl, and then extracted with EtOAc (80 mL, 2 ×) . The combined organic phase was concentrated to afford the crude compound 1226-2 without further purification. MS m/z (ESI) : 219.0 [M+H] ⁺.

Step 2

To a solution of compound 1226-2 (9.6 g, 44.0 mmol) in acetonitrile (100 mL) were added K₂CO₃ (12.2 g, 88.3 mmol) and methyl iodide (7.2 mL, 88.8 mmol) at room temperature. The reaction solution was heated to 80 ℃ for 18 hours. After the reaction was completed, the reaction mixture was cooled and quenched with water (50 mL) . The aqueous solution was extracted with EtOAc (100 mL, 2 ×) . The combined organic phase was dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (EtOAc/petroleum ether = 1/20) to afford the desired compound 1226-3 (5.7 g, 22.6 mmol, 55.8%yield) as a colorless liquid. MS m/z (ESI) : 233.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 10.44 (s, 1H) , 8.23 –8.14 (m, 2H) , 8.04 (d, J = 8.0 Hz, 1H) , 3.94 (s, 3H) .

Step 3

To a solution of compound 1226-3 (750 mg, 3.23 mmol) in toluene (15 mL) were added compound 1226-4 (650 μL, 4.14 mmol) , anhydrous MgSO₄ (2.0 g) , and catalyst (CAS: 791616-63-2, 240 mg, 0.32 mmol) at room temperature. After stirring for 18 hours, the reaction mixture was filtered through a pad of celite. The filtrate was collected and subsequently concentrated, and the resulting residue was purified by silica gel column chromatography (EtOAc/petroleum ether = 1/1) to afford the desired compound 1226-5 (160 mg, 0.47 mmol, 14.5%yield) as a white solid.

Step 4

To a solution of compound 1226-5 (120 mg, 0.35 mmol) in CH₂Cl₂ (7.5 mL) was added TFA (1.5 mL, 19.6 mmol) at room temperature. After stirring for 0.5 hours, the reaction mixture was concentrated to afford the crude intermediate 1226-6 without further purification. MS m/z (ESI) : 243.0 [M+H] ⁺.

Intermediate Example 2: Synthesis of intermediate 1226-7

Step 1

To a solution of compound 1226-8 (5.0 g, 21.7 mmol) in DMSO (150 mL) were added bis (pinacolato) diboron (16.5 g, 65.2 mmol) , KOAc (6.40 g, 65.2 mmol) , and Pd (dppf) Cl₂ (3.18 g, 4.35 mmol) at room temperature under N₂. The reaction solution was heated to 100 ℃ and stirred for 16 hours. After the reaction was completed, the reaction mixture was cooled and filtered. The filtrate was treated with water (500 mL) , and the aqueous solution was extracted with EtOAc (300 mL) . The organic phase was collected and washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (EtOAc/petroleum ether = 1/10) to afford the desired compound 1126-9 (5.9 g, 21.3 mmol, 98.0%yield) as a yellow solid.

Step 2

To a solution of compound 1226-9 (700 mg, 2.53 mmol) and compound 1226-10 (564 mg, 3.03 mmol) in 1, 4-dioxane (5 mL) and water were added X-Phos Pd G3 (214 mg, 0.25 mmol) , X-Phos (241 mg, 0.51 mmol) , and K₃PO₄ (1.07 g, 5.05 mmol) at room temperature under N₂. The reaction solution was heated to 100 ℃ and stirred for 16 hours. After the reaction was completed, the reaction mixture was cooled and filtered. The filtrate was collected and treated with water (20 mL) , and the aqueous solution was extracted with EtOAc (30 mL) . The organic phase was collected washed with brine (20 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (EtOAc/petroleum ether = 1/10) to afford the desired compound 1226-11 (630 mg, 2.46 mmol, 97.3%yield) as a yellow solid.

Step 3

A solution of compound 1226-11 (90 mg, 0.35 mmol) and lithium hydroxide (133 mg, 3.51 mmol) in MeOH (3 mL) and water (0.6 mL) was heated to 60 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was acidified to pH = 3 using 1 M aq. HCl and formed a suspension. The suspension was filtered, and the filter cake was washed with water (3 mL) and then dried under reduced pressure to obtain intermediate 1226-7 (54.0 mg, 0.22 mmol, 63.5%yield) as a white solid.

Intermediate Example 3: Synthesis of intermediate 1235-4

Step 1

Preparation of compound 1235-7 Grignard reagent:

To a solution of compound 1235-7 (3.0 g, 11.0 mmol) in THF (11 mL) was slowly added Mg dust (0.8 g, 32.9 mmol) at 25 ℃ under N₂. The reaction mixture was stirred for 0.5 hours to form the compound 1235-7 Grignard reagent (3 M) .

To a solution of compound 1235-6 (1.0 g, 4.65 mmol) in THF (20 mL) was added compound 1235-7 Grignard reagent (3 M, 18 mL) dropwise at 0 ℃ under N₂. The reaction solution was heated to 25 ℃ and stirred for 1 hour. After the reaction was completed, the reaction mixture was poured into saturated aq. NH₄Cl (200 mL) . The aqueous solution was extracted with EtOAc (200 mL, 3 ×) , and the combined organic phase was washed with brine (500 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (20-40%EtOAc in petroleum ether) to afford the desired intermediate 1235-1 (1.40 g, 3.87 mmol, 83.4%yield) as a yellow solid. MS m/z (ESI) : 362.2 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ: 7.60 –7.55 (m, 2H) , 7.49 –7.44 (m, 2H) , 7.38 –7.27 (m, 5H) , 4.67 –4.55 (m, 2H) , 3.74 (d, J = 10.8 Hz, 1H) , 3.60 (d, J = 10.8 Hz, 1H) , 1.31 –1.19 (m, 2H) , 0.96 –0.85 (m, 1H) , 0.44 –0.33 (m, 1H) .

Step 2

To a solution of compound 1235-1 (200 mg, 0.55 mmol) in CHCl₃ (5 mL) were added compound 1235-2 (10 μL, 0.13 mmol) and N-Bromosuccinimide (NBS, 246 mg, 1.38 mmol) at 25 ℃ under N₂. After stirring for 16 hours, the reaction mixture was concentrated to afford the crude compound 1235-3 (501 mg, 0.53 mmol, 96.5%) as a brown oil without purification. MS m/z (ESI) : 376.2 [M+H] ⁺.

Step 3

To a solution of compound 1235-3 (501 mg, 0.53 mmol) in methanol (20 mL) was added K₂CO₃ (153 mg, 1.11 mmol) at 25 ℃ under N₂. After stirring for 2 hours, the reaction mixture was filtered. The filter cake was washed with acetonitrile (10 mL, 3 ×) . The filtrate was collected, combined and concentrated. The residue was purified by silica gel column chromatography (0-10%methanol in CH₂C₂) to afford the desired intermediate 1235-4 (60 mg, 0.22 mmol, 41.4%yield) as a brown solid. MS m/z (ESI) : 272.0 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d₆) δ: 7.91 (s, 1H) , 7.71 (d, J = 8.4 Hz, 2H) , 7.61 (d, J = 8.4 Hz, 2H) , 5.85 (s, 1H) , 3.69 (d, J = 10.4 Hz, 1H) , 3.53 (d, J = 10.4 Hz, 1H) , 0.96 –0.75 (m, 3H) , 0.08 –0.02 (m, 1H) .

Intermediate Example 4: Synthesis of intermediate 1235-5

Step 1

To a solution of intermediate 1226-7 (500 mg, 2.06 mmol) in DMSO (10 mL) were added Et₃N (TEA, 0.6 mL, 4.33 mmol) and diphenylphosphoryl azide (DPPA, 0.90 mL, 4.14 mmol) at room temperature under N₂. The reaction solution was heated to 100 ℃ and stirred for 4 hours. After the reaction was completed, the reaction mixture was cooled and poured into water (20 mL) . The aqueous solution was extracted with EtOAc (20 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-10%methanol in CH₂C₂) to afford the desired compound 1235-8 (56.0 mg, 0.21 mmol, 10.2%yield) as a yellow solid. MS m/z (ESI) : 214.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.16 (s, 1H) , 7.43 (d, J = 2.4 Hz, 1H) , 7.35 (d, J = 8.8 Hz, 1H) , 6.90 (dd, J = 8.8, 2.4 Hz, 1H) , 6.59 (s, 2H) , 5.08 (s, 2H) , 4.33 (s, 3H) .

Step 2

To a solution of compound 1235-8 (56.0 mg, 0.21 mmol) in acetonitrile (2 mL) were added p-TsOH (54 mg, 0.31 mmol) , n-Bu₄NBr (TBAB, 170 mg, 0.53 mmol) , CuBr (6.0 mg, 0.03 mmol) , and t-BuONO (16.0 mg, 0.16 mmol) at room temperature under N₂. After stirring for 2 hours, the reaction mixture was concentrated. The residue was purified by silica gel column chromatography (0-5%methanol in CH₂C₂) to afford the desired intermediate 1235-5 (155 mg, 0.13 mmol, 85.2%yield) as a brown solid. MS m/z (ESI) : 277.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ: 8.32 (d, J = 2.4 Hz, 1H) , 8.30 (s, 1H) , 7.65 (dd, J = 2.4, 8.8 Hz, 1H) , 7.53 (d, J = 8.8 Hz, 1H) , 7.32 (s, 2H) , 4.39 (s, 3H) .

Intermediate Example 5: Synthesis of intermediate 1236-10

Route 1

Step 1

To a solution of compound 1236-1 (7.0 g, 34.8 mmol) in toluene (70 mL) was added NaH (2.1 g, 52.5 mmol, 60%purity) at 0 ℃. The reaction mixture was then heated to 80 ℃, and dimethyl sulfate (3.6 mL, 38.0 mmol) was added. After stirring for 3 hours, the reaction solution was cooled to room temperature and the reaction was quenched with saturated aq. NH₄Cl (100 mL) . The reaction mixture was extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-15%EtOAc in petroleum ether) to afford the desired compound 1236-2 (3.28 g, 15.2 mmol, 43.8%yield) as a red solid. MS m/z (ESI) : 216.0 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ 7.37 –7.30 (m, 2H) , 7.02 (s, 1H) , 3.58 (s, 2H) , 3.25 (s, 3H) .

Step 2

A solution of NaH (2.4 g, 50.0 mmol, 60%purity) in toluene (30 mL) was stirred at 25 ℃for 10 minutes. Then a solution of compound 1236-2 (3.1 g, 14.4 mmol) and dimethyl carbonate (2.5 mL, 29.7 mmol) in toluene (30 mL) was added dropwise into the above-mentioned NaH solution. After stirring at 25 ℃ for 30 hours, the reaction mixture was quenched with saturated aq. NH₄Cl (50 mL) . The reaction mixture was extracted with EtOAc (30 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-20%EtOAc in petroleum ether) to afford the desired compound 1236-3 (1.35 g, 4.94 mmol, 34.3%yield) as a yellow solid. MS m/z (ESI) : 274.0 [M+H] ⁺.

Step 3

A solution of compound 1236-3 (1.1 g, 4.03 mmol) , compound 1236-5 (1.1 g, 4.83 mmol) , and K₂CO₃ (1.1 g, 8.05 mmol) in acetonitrile (20 mL) was stirred at 80 ℃ for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and filtered. The resulting filtrate was concentrated and purified by silica gel column chromatography (0-15%EtOAc in petroleum ether) to afford the desired compound 1236-6 (1.18 g, 3.04 mmol, 75.6%yield) as a white solid. MS m/z (ESI) : 389.0 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ7.39 –7.33 (m, 6H) , 7.15 –7.07 (m, 2H) , 4.76 (s, 2H) , 3.35 (d, J = 18.0 Hz, 1H) , 3.31 (s, 3H) , 2.97 (d, J = 18.0 Hz, 1H) .

Step 4

To a solution of compound 1236-6 (1.3 g, 3.35 mmol) in CH₂Cl₂ (15 mL) was added LiEt₃BH (4 mL, 4.00 mmol, 1 M in THF) dropwise at -60 ℃ under N₂. After stirring for 2 hours, the reaction mixture was heated to room temperature and quenched with saturated aq. NH₄Cl (20 mL) . The reaction mixture was extracted with EtOAc (20 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-20%EtOAc in petroleum ether) to afford the desired compound 1236-7 (0.69 g, 1.77 mmol, 62.3%yield) as a white solid. MS m/z (ESI) : 413.2 [M+Na] ⁺. ¹H NMR (400 MHz, CDCl₃) δ 7.46 –7.28 (m, 7H) , 7.06 (s, 1H) , 5.16 (s, 0.7H) , 5.07 (d, J = 14.4 Hz, 0.3 H) , 4.95 (d, J = 14.8 Hz, 0.7H) , 4.88 (s, 0.3H) , 4.35 (d, J = 14.8 Hz, 0.7H) , 4.19 (d, J = 14.4 Hz, 0.3H) , 3.28 (s, 1H) , 3.24 (s, 2H) , 3.20 (d, J = 16.8 Hz, 0.3H) , 2.93 (d, J = 17.2 Hz, 0.7H) , 2.80 (d, J = 16.8 Hz, 0.7H) , 2.55 (d, J = 17.2 Hz, 0.3H) .

Step 5

To a solution of compound 1236-7 (0.69 g, 1.77 mmol) and Et₃SiH (1.4 mL, 8.84 mmol) in CH₂Cl₂ (8 mL) was added BF₃·Et₂O (0.6 mL, 4.86 mmol) at -60 ℃ under N₂. The reaction solution was heated to -20 ℃ and stirred for 5 hours. After the reaction was completed, the reaction mixture was heated to room temperature and quenched with water (50 mL) . The aqueous solution was extracted with EtOAc (30 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-20%EtOAc in petroleum ether) to afford the desired compound 1236-8 (576 mg, 1.54 mmol, 87.1%yield) as a white solid. MS m/z (ESI) : 375.2 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ 7.39 –7.27 (m, 6H) , 7.21 (d, J = 7.6 Hz, 1H) , 7.04 (s, 1H) , 4.72 (d, J = 14.8 Hz, 1H) , 4.43 (d, J = 14.8 Hz, 1H) , 3.72 (d, J = 9.6 Hz, 1H) , 3.25 (d, J = 10.0 Hz, 1H) , 3.24 (s, 3H) , 3.10 (d, J = 16.8 Hz, 1H) , 2.58 (d, J = 16.8 Hz, 1H) .

Step 6

A solution of compound 1236-8 (250 mg, 0.67 mmol) , NBS (180 mg, 1.01 mmol) , and 2, 2’-Azobis (2-methylpropionitrile) (AIBN, 18 mg, 0.11 mol) in CHCl₃ (3 mL) was stirred at 130 ℃ for 12 hours under N₂. After the reaction was completed, the reaction mixture was cooled to room temperature and treated with water (50 mL) . The aqueous solution was extracted with EtOAc (30 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated to afford the crude compound 1236-9 (259 mg, 0.67 mmol, 100%yield) as an off-white solid without further purification. MS m/z (ESI) : 389.1 [M+H] ⁺.

Step 7

A solution of compound 1236-9 (259 mg, 0.67 mmol) and K₂CO₃ (185 mg, 1.34 mmol) in methanol (3 mL) was stirred at 20 ℃ for 2 hours. After the reaction was completed, the reaction mixture was treated with water (50 mL) , and the aqueous solution was extracted with EtOAc (30 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-30%EtOAc in petroleum ether) to afford the crude intermediate 1236-10 (75 mg, 0.26 mmol, 39.5%yield) as an off-white solid. MS m/z (ESI) : 285.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.04 (s, 1H) , 7.63 (d, J = 8.0 Hz, 1H) , 7.45 (d, J = 7.6 Hz, 1H) , 7.39 (s, 1H) , 3.50 (d, J = 9.6 Hz, 1H) , 3.41 (d, J = 10.0 Hz, 1H) , 3.19 (s, 3H) , 2.53 –2.51 (m, 2H) .

Route 2

Step 1

To a solution of compound 1264-2 (5.0 g, 11.8 mmol) in methanol (50 mL) and water (10 mL) were added Fe powder (6.60 g, 118) and NH₄Cl (6.3 g, 118 mmol) at room temperature under N₂. The reaction solution was heated to 60 ℃ and stirred for 12 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, filtered, and concentrated. The residue was added with water (100 mL) , and the aqueous solution was extracted with CH₂Cl₂ (30 mL, 3 ×) . The combined organic phase was washed with brine (50 mL, 2 ×) , dried over Na₂SO₄, filtered, and concentrated. The crude residue was purified by silica gel column chromatography (methanol/CH₂Cl₂ = 1/20) to afford the desired compound 1236-11 (4.0 g, 11.1 mmol, 93.8%yield) as a yellow solid. MS m/z (ESI) : 361.0 [M+H] ⁺.

Step 2

To a solution of compound 1236-11 (5.0 g, 13.4 mmol) in DMF (50 mL) was added NaH (0.67 g, 27.75 mmol, 60%purity) at 0 ℃. After the solution was stirred for 0.5 hours, methyl iodide (1.97 g, 13.9 mmol) was added. The reaction solution was stirred at 0 ℃ for 3 hours and then quenched with water (100 mL) . The aqueous solution was extracted EtOAc (20 mL, 2 ×) . The combined organic phase was washed with brine (40 mL, 2 ×) , dried over Na₂SO₄, filtered, and concentrated to afford the desired compound 1236-8 (4.80 g, 12.8 mmol, 92.4%yield) as a yellow oil. MS m/z (ESI) : 375.0 [M+H] ⁺.

Step 3

A solution of compound 1236-8 (370 mg, 1.06 mmol) , AIBN (28.0 mg, 0.17 mmol) , and NBS (274 mg, 1.60 mmol) in chlorobenzene (5 mL) was stirred at 130 ℃ for 12 hours under N₂. Then the reaction was cooled and treated with water (0.5 mL) . The reaction was heated again to130 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and concentrated. The residue was treated with methanol (5 mL) , water (1 mL) , and LiOH (281 mg, 6.70 mmol) . The reaction solution was heated to 60 ℃stirred for 12 hours. After the reaction was completed, the reaction mixture was cooled and concentrated. The residue was purified by silica gel column chromatography (methanol/CH₂Cl₂ = 1/20) to afford the desired intermediate 1236-10 (60 mg, 0.21 mmol, 31.6%yield) as a yellow solid. MS m/z (ESI) : 285.0 [M+H] ⁺.

Intermediate Example 6: Synthesis of intermediate 1236-15

Step 1

A solution of compound 1236-17 (10 g, 53.9 mmol) and NBS (11.5 g, 64.7 mmol) in THF (100 mL) was stirred at room temperature for 10 hours. The reaction mixture was quenched with saturated aq. NH₄Cl (300 mL) . The aqueous solution was extracted with EtOAc (100 mL, 3 ×) . The combined organic phase was washed with brine (300 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-33%EtOAc in petroleum ether) to afford the desired compound 1236-18 (8.0 g, 30.3 mmol, 56.1%yield) . MS m/z (ESI) : 263.8, 265.8 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.91 (s, 1H) , 6.86 (s, 1H) , 6.39 (s, 2H) , 3.76 (s, 3H) .

Step 2

Following the same procedures as in step 1 of Intermediate Example 2, compound 1236-19 was obtained (4.0 g, 12.8 mmol, 42.5%yield) . MS m/z (ESI) : 312.0, 314.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.02 (s, 1H) , 6.74 (s, 1H) , 6.38 (s, 2H) , 3.75 (s, 3H) , 1.30 (s, 12H) .

Step 3

Following the same procedures as in step 2 of Intermediate Example 2, compound 1236-20 was obtained (0.5 g, 1.72 mmol, 53.6%yield) . MS m/z (ESI) : 291.0, 293.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.70 (s, 1H) , 8.28 (s, 1H) , 7.60 (s, 1H) , 7.54 (s, 2H) , 4.39 (s, 3H) , 3.90 (s, 3H) .

Step 4

Following the same procedures as in step 3 of Intermediate Example 2, compound 1236-21 was obtained (0.8 g, 2.89 mmol, 93.4%yield) . MS m/z (ESI) : 277.0, 279.0 [M+H] ⁺.

Step 5

To a solution of compound 1236-21 (3.0 g, 10.3 mmol) and PMBCl (3.3 mL, 23.2 mmol) in DMF (30 mL) was added NaH (1.70 g, 42.5 mmol, 60%purity) at 0 ℃. The reaction solution was heated to 25 ℃ and stirred for 12 hours. After the reaction was completed, the reaction mixture was poured into aq. 1 M HCl (100mL) , and the quenched solution was stirred for 10 minutes. After white solids were precipitated, the suspension was filtered. The collected filter cake was washed with acetonitrile (50 mL) and dried to afford the desired compound 1236-11 (3.53 g, 6.83 mmol, 66.1%yield) as a white solid. MS m/z (ESI) : 517.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.78 (s, 1H) , 7.83 (s, 1H) , 7.71 (s, 1H) , 7.26 (d, J = 8.0 Hz, 4H) , 6.89 (d, J = 8.8 Hz, 4H) , 5.03 (s, 4H) , 4.37 (s, 3H) , 3.71 (s, 6H) .

Step 6

To a solution of compound 1236-11 (3.53 g, 6.83 mmol) and Et₃N in t-BuOH (30 mL) was added DPPA (2.3 mL, 10.6 mmol) . The reaction was heated to 80 ℃ and stirred for 12 hours. After completion, the reaction mixture was cooled and diluted with water (100 mL) . The mixture was then extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-15%EtOAc in petroleum ether) to afford the desired compound 1236-13 (2.91 g, 4.95 mmol, 72.4%yield) MS m/z (ESI) : 588.4 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.74 (s, 1H) , 8.42 (s, 1H) , 7.80 (s, 1H) , 7.73 (s, 1H) , 7.26 (d, J = 8.8 Hz, 4H) , 6.88 (d, J = 8.8 Hz, 4H) , 4.98 (s, 4H) , 4.33 (s, 3H) , 3.71 (s, 6H) , 1.48 (s, 9H) .

Step 7

To a solution of compound 1236-13 (2.91 g, 4.95 mmol) in CH₂Cl₂ (15 mL) was added TFA (5 mL, 67.1 mmol) at 20 ℃. After stirring for 2 hours, the reaction mixture was poured into the saturated aq. NaHCO₃ (50 mL) . The aqueous was extracted with CH₂Cl₂ (30 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-20%EtOAc in petroleum ether) to afford the desired compound 1236-14 (1.3 g, 2.66 mmol, 53.9%yield) as a pink solid.

Step 8

Following the same procedures as in step 2 of Intermediate Example 4, intermediate 1236-15 was obtained (260 mg, 0.47 mmol, 18.4%yield) as a yellow solid. MS m/z (ESI) : 551.3, 553.3 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.36 (s, 1H) , 7.94 (s, 1H) , 7.71 (s, 1H) , 7.25 (d, J = 8.4 Hz, 4H) , 6.86 (d, J = 8.8 Hz, 4H) , 4.98 (s, 4H) , 4.39 (s, 3H) , 3.79 (s, 6H) .

Intermediate Example 7: Synthesis of intermediate 1241-7

Step 1

To a solution of compound 1235-1 (500 mg, 1.38 mmol) in DMF (7 mL) were added pyridine (0.22 mL, 2.73 mmol) and SOCl₂ (0.16 mL, 2.21 mmol) at 0 ℃ under N₂. After heating the solution to 25 ℃ and stirring for 2 hours, the reaction mixture was poured into water (50 mL) . The aqueous solution was extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-10%EtOAc in petroleum ether) to afford the desired compound 1241-4 (204 mg, 0.53 mmol, 38.7%yield) as a yellow oil. MS m/z (ESI) : 344.2 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ: 7.51 (d, J = 8.0 Hz, 2H) , 7.40 –7.29 (m, 5H) , 7.20 (d, J = 8.0 Hz, 2H) , 6.83 (s, 1H) , 4.79 (s, 2H) , 1.74 –1.69 (m, 2H) , 1.68 –1.60 (m, 2H) .

Step 2

A solution of compound 1241-4 (154 mg, 0.45 mmol) and Pd (OH) ₂ on charcoal (120 mg, 10%purity) in ethanol (20 mL) was degassed by purging H₂ for 3 times at 0 ℃. The reaction solution was stirred at 25 ℃ under H₂ for 3 hours. After the reaction was completed, the reaction mixture was filtered, and the filter cake was washed with methanol (10 mL, 3 ×) . The combined filtrate was concentrated. The residue was purified by silica gel column chromatography (0-30%EtOAc in petroleum ether) to afford the desired compound 1241-5 (108 mg, 0.31 mmol, 69.7%yield) as a yellow oil. MS m/z (ESI) : 346.0 [M+H] ⁺.

Step 3

To a solution of compound 1241-5 (135 mg, 0.39 mmol) in CHCl₃ (3.5 mL) were added compound 1235-2 (6 μL, 0.08 mmol) and NBS (174 mg, 0.98 mmol) at 25 ℃ under N₂. After stirring for 16 hours, the reaction mixture was poured into water (40 mL) , and the aqueous solution was extracted with EtOAc (40 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-10%methanol in CH₂Cl₂) to afford the desired intermediate 1241-7 (8.0 mg, 0.11 mmol, 28.1%yield) as a yellow solid. MS m/z (ESI) : 256.0 [M+H] ⁺; ¹H NMR (500 MHz, DMSO-d₆) δ: 7.85 (s, 1H) , 7.69 (d, J = 8.0 Hz, 2H) , 7.43 (d, J = 8.0 Hz, 2H) , 3.84 –3.73 (m, 1H) , 3.69 –3.56 (m, 1H) , 3.37 –3.34 (m, 1H) , 0.94 –0.89 (m, 1H) , 0.80 –0.73 (m, 2H) , 0.17 –0.09 (m, 1H) .

Intermediate Example 8: Synthesis of intermediate 1250-1

Step 1

Following the same procedures as in step 1 of Intermediate Example 6, compound 1250-2 was converted to compound 1250-3 (7.8 g, 31.5 mmol, 88.7%yield) as an off-white solid. MS m/z (ESI) : 247.8, 249.8 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Intermediate Example 6, compound 1250-3 was converted to compound 1250-4 (2.2 g, 7.45 mmol, 92.5%yield) as an off-white solid. MS m/z (ESI) : 296.0 [M+H] ⁺.

Step 3

Following the same procedures as in step 3 of Intermediate Example 6, compound 1250-4 was converted to compound 1250-5 (300 mg, 1.09 mmol, 32.3%yield) as an off-white solid. MS m/z (ESI) : 275.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.72 (d, J = 8.0 Hz, 1H) , 8.27 (s, 1H) , 7.55 (s, 2H) , 7.28 (d, J = 13.2 Hz, 1H) , 4.39 (s, 3H) , 3.89 (s, 3H) .

Step 4

Following the same procedures as in step 3 of Intermediate Example 2, compound 1250-5 was converted to intermediate 1250-1 (200 mg, 0.77 mmol, 70.3%yield) as an off-white solid. MS m/z (ESI) : 261.0 [M+H] ⁺.

Intermediate Example 9: Synthesis of intermediate 1252-4

Step 1

Compounds 1226-3 and Compounds 1252-2 were used to follow a procedure same as described in step 3 of Intermediate Example 1 to give compound 1250-3 (500 mg, 1.40 mmol, 32.6%yield) as colorless oil. MS m/z (ESI) : 357.4 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.04 (dd, J = 1.6, 8.0 Hz, 1H) , 7.98 (s, 1H) , 7.93 (d, J = 8.0 Hz, 1H) , 6.25 (s, 1H) , 4.25 –4.14 (m, 1H) , 3.67 –3.56 (m, 1H) , 3.32 –3.23 (m, 1H) , 2.98 –2.84 (m, 1H) , 1.93 –1.71 (m, 2H) , 1.32 (s, 9H) .

Step 2

Following the same procedures as in step 4 of Intermediate Example 1, compound 1250-3 was converted to crude intermediate 1250-4 without further purification as colorless oil. MS m/z (ESI) : 357.4 [M+H] ⁺.

Intermediate Example 10: Synthesis of intermediate 1264-6

Step 1

To a solution of compound 1264-1 (20.0 g, 95.7 mmol) in THF (500 mL) was added KHMDS (143 mL, 143 mmol, 1.0 M) at -78 ℃ under N₂. After the reaction solution was stirred for 1 hour, compound 1264-A (24.5 g, 105 mmol) was added. The resulting reaction mixture was stirred at -78 ℃ for 2 hours, heated to room temperature, and stirred for another 16 hours. After the reaction was completed, the reaction was quenched with saturated aq. NH₄Cl (300 mL) . Then the aqueous was diluted with water (200 mL) and extracted with EtOAc (200 mL, 2 ×) . The combined organic phase was washed with brine (200 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-25%EtOAc in petroleum ether) to afford the desired compound 1264-2 (18.0 g, 42.0 mmol, 44.6%yield) as yellow oil. MS m/z (ESI) : 423.2 [M+H] ⁺.

Step 2

To a solution of compound 1264-2 (3.6 g, 8.52 mmol) , CaCl₂ (1.89 g, 17.1 mmol) in THF (20 mL) /water (10 mL) was added NaBH₄ (0.97 g, 25.6 mmol) in portions at 0 ℃ under N_2. The reaction solution was first stirred for 0.5 hours and warmed to room temperature for another 2 hours. After the reaction was completed, the reaction was quenched with water, and the aqueous solution was extracted with EtOAc (50 mL, 2 ×) . The combined organic phase was washed with brine (70 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-20%EtOAc in petroleum ether) to afford the desired compound 1264-3 (1.75 g, 4.44 mmol, 52.0%yield) as a white solid. MS m/z (ESI) : 395.0 [M+H] ⁺.

Step 3

A solution of compound 1264-3 (1.75 g, 4.44 mmol) and Pd on charcoal (200 mg, 10%purity) in methanol (20 mL) was degassed through H₂ for 3 times at room temperature. The reaction solution was stirred under H₂ for 2 hours. After the reaction was completed, the reaction mixture was filtered and concentrated to afford the desired compound 1264-4 (1.46 g, 3.95 mmol, 89.0%yield) as yellow oil. MS m/z (ESI) : 365.2 [M+H] ⁺.

Step 4

To a solution of compound 1264-4 (1.46 g, 3.95 mmol) in acetonitrile (10 mL) were added copper iodide (1.5 g, 7.90 mmol) and Amyl nitrile (555 mg, 4.74 mmol) at 0 ℃. The reaction solution was first stirred for 0.5 hours and warmed to room temperature for another 2 hours. After the reaction was completed, the reaction mixture was filtered and concentrated. The residue was purified by silica gel column chromatography (0-25%EtOAc in petroleum ether) to afford the desired compound 1264-5 (513 mg, 1.44 mmol, 36.0%yield) as a white solid. MS m/z (ESI) : 348.0 [M+H] ⁺.

Step 5

A solution of compound 1264-5 (370 mg, 1.06 mmol) , AIBN (28.0 mg, 0.17 mmol) , and NBS (274 mg, 1.60 mmol) in chlorobenzene (5 mL) was stirred at 130 ℃ for 16 hours under N₂. Then water (0.5 mL) was added, the reaction solution was stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and concentrated. The residue was treated with methanol (5 mL) and K₂CO₃ (294 mg, 2.13 mmol) . The reaction solution was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was filtered and concentrated. The residue was purified by silica gel column chromatography (methanol/CH₂Cl₂ = 1/30) to afford the desired intermediate 1264-6 (100 mg, 0.39 mmol, 36.0%yield) as a white solid. MS m/z (ESI) : 257.9 [M+H] ⁺.

Intermediate Example 11: Synthesis of intermediate 1267-10

Step 1

Following the same procedures as in step 5 of Intermediate Example 6, compound 1267-2 was obtained (6.0 g, 11.7 mmol, 32.0%yield) as a colorless liquid. MS m/z (ESI) : 515.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 3 of Intermediate Example 2, compound 1267-3 was obtained (1.0 g, 1.93 mmol, 41.1%yield) as a solid. MS m/z (ESI) : 501.2 [M+H] ⁺.

Step 3

Following the same procedures as in step 6 of Intermediate Example 6, compound 1267-4 was obtained (2.8 g, 4.90 mmol, 49.0%yield) as a white solid. MS m/z (ESI) : 572.2 [M+H] ⁺.

Step 4

Following the same procedures as in step 4 of Intermediate Example 1, compound 1267-5 was obtained (2.0 g, 1.24 mmol, 97.1%yield) as a white solid. MS m/z (ESI) : 472.2 [M+H] ⁺.

Step 5

Following the same procedures as in step 2 of Intermediate Example 4, compound 1267-10 was obtained (200 mg, 0.36 mmol, 49.0%yield) as a white solid. MS m/z (ESI) : 535.2, 537.2 [M+H] ⁺.

Intermediate Example 12: Synthesis of intermediate 1268-8 and 1268-11

Step 1

Following the same procedures as in step 2 of Intermediate Example 2, compound 1268-3 was obtained (3.0 g, 11.4 mmol, 75.0%yield) as a yellow solid. MS m/z (ESI) : 263.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 5 of Intermediate Example 6, compound 1268-4 was obtained (3.5 g, 6.96 mmol, 91.3%yield) as a yellow oil. MS m/z (ESI) : 503.2 [M+H] ⁺.

Step 3

Following the same procedures as in step 3 of Intermediate Example 2, compound 1268-5 was obtained (1.8 g, 3.68 mmol, 92.6%yield) as a yellow solid. MS m/z (ESI) : 489.2 [M+H] ⁺.

Step 4

Following the same procedures as in step 6 of Intermediate Example 6, compound 1268-6 was obtained (2.0 g, 3.57 mmol, 72.7%yield) as a yellow solid. MS m/z (ESI) : 560.3 [M+H] ⁺.

Step 5

Following the same procedures as in step 4 of Intermediate Example 1, compound 1268-7 was obtained (300 mg, 0.65 mmol, 73.1%yield) as a white solid. MS m/z (ESI) : 460.2 [M+H] ⁺.

Step 6

Following the same procedures as in step 2 of Intermediate Example 4, compound 1268-8 was obtained (80.0 mg, 0.15 mmol, 58.5%yield) . MS m/z (ESI) : 523.1, 525.1 [M+H] ⁺.

Step 7

Following the same procedures as in step 2 of Intermediate Example 3, compound 1268-11 was obtained.

Intermediate Example 13: Synthesis of intermediate 1275-10

Step 1

To a solution of compound 1264-3 (2.60 g, 6.59 mmol) in CH₂Cl₂ (20 mL) were added imidazole (670 mg, 6.89 mmol) and TBSCl (1.99 g, 13.2 mmol) at room temperature. After stirring for 3 hours, the reaction solution was quenched with water, and the aqueous solution was extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (70 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1275-4 (2.5 g, 5.22 mmol, 79.2%yield) as a white solid. MS m/z (ESI) : 509.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 3 of Intermediate Example 10, compound 1275-5 was obtained (1.9 g, 4.05 mmol, 82.0%yield) as a yellow oil. MS m/z (ESI) : 479.2 [M+H] ⁺.

Step 3

Following the same procedures as in step 4 of Intermediate Example 10, compound 1275-6 was obtained (330 mg, 0.56 mmol, 62.0%yield) as a yellow oil. MS m/z (ESI) : 590.2 [M+H] ⁺.

Step 4

To a solution of compound 1275-6 (210 mg, 0.36 mmol) in DMF (2 mL) and MeOH (1 mL) were added Pd (dppf) Cl₂ (29.0 mg, 0.04 mmol) and Et₃N (72.0 mg, 0.71 mmol) at room temperature under CO. The reaction solution was heated to 80 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled, filtered, and concentrated. The residue was purified by silica gel column chromatography (16%EtOAc in petroleum ether) to afford the desired compound 1275-7 (175 mg, 0.34 mmol, 69.4%yield) as a yellow oil. MS m/z (ESI) : 522.2 [M+H] ⁺.

Step 5

Following the same procedures as in step 2 of Intermediate Example 10, compound 1275-8 was obtained (230 mg, 0.47 mmol, 43.0%yield) as a white solid. MS m/z (ESI) : 494.2 [M+H] ⁺.

Step 6

To a solution of compound 1275-8 (200 mg, 0.41 mmol) in toluene (5 mL) was added conc. H₂SO₄ (0.1 mL) at room temperature. The reaction solution was heated to 100 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled and quenched with water, and the aqueous solution was extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (70 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (20%EtOAc in petroleum ether) to afford the desired compound 1275-9 (88.0 mg, 0.24 mmol, 60.0%yield) as a white solid. MS m/z (ESI) : 361.2 [M+H] ⁺.

Step 7

A solution of compound 1275-9 (80.0 mg, 0.24 mmol) and CAN (616 mg, 1.11 mmole) in acetonitrile (3 mL) and water (1.5 mL) was stirred at 100 ℃ for 16 hours. After the reaction was completed, the reaction was solution cooled, filtered, and concentrated. The residue was purified by RP-silica gel column chromatography (30%acetonitrile in 0.1%TFA of water) to afford the desired intermediate 1275-10 (18.0 g, 0.03 mmol, 15.0%yield) . MS m/z (ESI) : 375.1 [M+H] ⁺.

Intermediate Example 14: Synthesis of intermediate 1277-5

Step 1

Following the same procedures as in step 2 of Intermediate Example 6, compound 1277-2 was obtained (6.50 g, 21.8 mmol, 65.0%yield) as a white solid. MS m/z (ESI) : 298.0, 300.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Intermediate Example 2, compound 1277-4 was obtained (60.0 g, 0.23 mmol, 67.4%yield) as a white solid. MS m/z (ESI) : 265.0, 267.0 [M+H] ⁺.

Step 3

Following the same procedures as in step 5 of Intermediate Example 6, intermediate 1277-5 was obtained (20.0 mg, 0.04 mmol, 10.5%yield) as a white solid. MS m/z (ESI) : 505.0, 507.0 [M+H] ⁺.

Intermediate Example 15: Synthesis of intermediate 1281-3

Step 1

A solution of compound 1236-11 (200 mg, 0.60 mmol) , cyclopropyl boronic acid (463 mg, 8.91 mmol) , K₂CO₃ (230 mg, 1.71 mmol) , and Cu (OAc) ₂ (443 mg, 2.22 mmol) in 1, 2-dichloroethane (8 mL) was stirred at 70 ℃ for 16 hours. After the reaction was completed, the reaction solution was cooled and filtered. The residue was purified by silica gel column chromatography (17%EtOAc in petroleum ether) to afford the desired compound 1281-2 (100 g, 0.25 mmol, 45.2%yield) as a yellow oil. MS m/z (ESI) : 400.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 5 of Intermediate Example 10, intermediate 1281-3 was obtained (50.0 mg, 0.16 mmol, 14.0%yield) as a white solid. MS m/z (ESI) : 311.0 [M+H] ⁺.

Intermediate Example 16: Synthesis of intermediate 1285-4

Step 1

To a solution of compound 1285-1 (5.0 g, 22.0 mmol) in CH₂Cl₂ (50 mL) were added AcOH (4.0 mL, 73.0 mmol) and NIS (6.0 g, 26.6 mmol) at room temperature. After stirring for 16 hours, the reaction mixture was poured into water (200 mL) , and the aqueous solution was extracted with EtOAc (100 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1285-2 (4.3 g, 13.0 mmol, 53.3%yield) as a white solid. MS m/z (ESI) : 332.0, 334.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Intermediate Example 6, compound 1285-3 was obtained (1.30 g, 3.90 mmol, 30.2%yield) . MS m/z (ESI) : 332.0, 334.0 [M+H] ⁺.

Step 3

Following the same procedures as in step 2 of Intermediate Example 2, intermediate 1285-4 was obtained (330 mg, 1.10 mmol, 28.2%yield) . MS m/z (ESI) : 299.0, 301.0 [M+H] ⁺.

Intermediate Example 17: Synthesis of intermediate 1286-4

Step 1

Following the same procedures as in step 2 of Intermediate Example 6, compound 1286-2 was obtained (1.10 g, 4.51 mmol, 88.8%yield) as an off-white solid. MS m/z (ESI) : 245.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Intermediate Example 2, crude compound 1286-3 was obtained without further purification. MS m/z (ESI) : 299.0, 212.0 [M+H] ⁺.

Step 3

To a solution of compound 1286-3 (230 mg, 1.09 mmol) in water (10 mL) was added NaOH (435 mg, 10.9 mmol) at room temperature. After stirring for 16 hours, the reaction solution was adjusted to pH = 6 using formic acid, forming a suspension with brown solids. The solids were collected by filtration, followed by washing with water (10 mL) , and then dried over under vacuum to afford crude desired intermediate 1286-4 without further purification. MS m/z (ESI) : 231.0 [M+H] ⁺.

Intermediate Example 18: Synthesis of intermediate 1289-5

Step 1

Following the same procedures as in step 1 of Intermediate Example 6, compound 1289-2 was obtained (7.80 g, 31.5 mmol, 88.7%yield) as an off-white solid. MS m/z (ESI) : 247.8, 249.8 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Intermediate Example 6, compound 1289-3 was obtained (2.20 g, 7.45 mmol, 92.5%yield) as a white solid. MS m/z (ESI) : 296.0 [M+H] ⁺.

Step 3

Following the same procedures as in step 2 of Intermediate Example 2, compound 1289-4 was obtained (150 mg, 0.57 mmol, 30.4%yield) as a yellow solid. MS m/z (ESI) : 262.9 [M+H] ⁺.

Step 4

Following the same procedures as in step 3 of Intermediate Example 2, compound 1289-5 was obtained (90.0 mg, 0.36 mmol, 95.1%yield) as a yellow solid. MS m/z (ESI) : 248.9 [M+H] ⁺.

Intermediate Example 19: Synthesis of intermediate 1290-3

Step 1

To a solution of compound 1290-4 (20.0 g, 105 mmol) in CH₂Br₂ (150 mL) were added K₂HPO₄ (55.0 g, 316 mmol) and Pd (OAc) ₂ (2.40 g, 10.5 mmol) at room temperature under N₂. After heating to 130 ℃ and stirring for 5 days, the reaction mixture was cooled and filtered. The filtrate was concentrated, and the resulting residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1290-5 (6.50 g, 32.1 mmol, 24.5%yield) . ¹H NMR (400 MHz, CDCl₃) δ 8.06 (d, J = 8.4 Hz, 1H) , 7.93 –7.74 (m, 2H) , 5.42 (s, 2H) .

Step 2

To a solution of compound 1290-5 (13.0 g, 64.3 mmol) in CH₂Cl₂ (130 mL) was added DIBAL-H (70 mL, 70 mmol, 1 M) dropwise at –70 ℃ under N₂. After stirring for 2 hours, the reaction mixture was quenched with water (250 mL) , and the aqueous solution was extracted with EtOAc (150 mL, 3 ×) . The combined organic phase was washed with brine (500 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1290-6 (8.0 g, 39.2 mmol, 60.9%yield) . ¹H NMR (400 MHz, CDCl₃) δ 7.63 (d, J = 7.6 Hz, 1H) , 7.56 (d, J = 8.4 Hz, 2H) , 6.54 (d, J = 2.0 Hz, 1H) , 5.31 –5.26 (m, 1H) , 5.09 (t, J = 12.4 Hz, 1H) .

Step 3

To a solution of compound 1290-6 (6.0 g, 29.4 mmol) in CH₂Cl₂ (60 mL) were added TMSCN (8.75 g, 88.2 mmol) and BF₃·OEt₂ (6.26 g, 44.1 mmol) at room temperature under N₂. The reaction solution was cooled to –20 ℃ and stirred for 0.5 hours. After reaction was completed, the reaction solution was poured into water (100 mL) , and the aqueous solution was extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1290-7 (5.50 g, 25.8 mmol, 87.8%yield) . ¹H NMR (400 MHz, CDCl₃) δ 7.69 (d, J = 8.00 Hz, 1H) , 7.62 –7.53 (m, 2H) , 6.00 –5.90 (m, 1H) , 5.35 (dd, J = 12.8, 2.4 Hz, 1H) , 5.23 (d, J = 12.8 Hz, 1H) .

Step 4

To a solution of compound 1290-7 (5.50 g, 25.8 mmol) and allyl bromide (9.36 g, 77.4 mmol) in DMF (50 mL) was added NaH (2.06 g, 51.6 mmol, 60%purity) at room temperature. After stirring for 2 hours, the reaction was quenched with saturated aq. NH₄Cl (200 mL) , and the aqueous solution was extracted with EtOAc (50 mL, 3 ×) . The combined organic phase was washed with brine (100 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1290-8 (5.30 g, 20.9 mmol, 81.1%yield) . ¹H NMR (400 MHz, CDCl₃) δ 7.67 (d, J = 8.05 Hz, 1H) , 7.56 (s, 1H) , 7.50 (d, J = 8.0 Hz, 1H) , 5.85 –5.72 (m, 1H) , 5.33 –5.15 (m, 4H) , 3.02 –2.91 (m, 1H) , 2.89 –2.81 (m, 1H) .

Step 5

To a solution of compound 1290-8 (2.0 g, 7.90 mmol) and RuCl₃ (0.21 g, 0.79 mmol) in acetonitrile (10 mL) , THF (10 mL) , and water (10 mL) was added NaIO₄ (8.45 g, 39.5 mmol) in portions at room temperature. After stirring for 2 hours, the reaction solution was dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1290-1 (1.10 g, 4.06 mmol, 51.4%yield) . MS m/z (ESI) : 270.0 [M-H] ^-.

Step 6

A solution of compound 1290-1 (750 mg, 2.77 mmol) and Ranny Ni (81.0 mg, 1.38 mmol) in 0.1%ammonia solution (in methanol, 10 mL) under H₂ was stirred for 12 hours. After the reaction was completed, the reaction mixture was filtered and concentrated to afford the desired compound 1290-2 (620 mg, 2.25 mmol, 81.5%yield) . MS m/z (ESI) : 275.0 [M+H] ⁺.

Step 7

A solution of compound 1290-2 (120 mg, 0.44 mmol) , Et₃N (132 mg, 1.31 mmol) , and HATU (249 mg, 0.65 mmol) in CH₂Cl₂ (2 mL) was stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was quenched with water (10 mL) , and the aqueous solution was extracted with CH₂Cl₂ (5 mL, 3 ×) . The combined organic phase was washed with brine (10 mL, 3 ×) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (50%EtOAc in petroleum ether) to afford the desired intermediate 1290-3 (100 mg, 0.39 mmol, 89.2%yield) . MS m/z (ESI) : 258.0 [M+H] ⁺.

Intermediate Example 20: Synthesis of intermediate 1295-5

Step 1

Following the same procedures as in step 1 of Intermediate Example 16, compound 1295-2 was obtained (2.38 g, 5.34 mmol, 99.0%yield) . MS m/z (ESI) : 313.8 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Intermediate Example 6, compound 1295-3 was obtained (4.10 g, 13.1 mmol, 82.0%yield) as a white solid. MS m/z (ESI) : 312.0 [M+H] ⁺.

Step 3

Following the same procedures as in step 2 of Intermediate Example 2, compound 1295-4 was obtained (140 mg, 0.50 mmol, 31.0%yield) . MS m/z (ESI) : 284.0 [M+H] ⁺.

Step 4

Following the same procedures as in step 5 of Intermediate Example 6, intermediate 1295-5 was obtained (143 mg, 0.28 mmol, 59.0%yield. MS m/z (ESI) : 519.2 [M+H] ⁺.

Intermediate Example 21: Synthesis of intermediate 1297-7

Step 1

To a solution of compound 1297-1 (9.0 g, 39.1 mmol) in CH₂Cl₂ (450 mL) was added DAST (31.5 g, 196 mmol) at 0 ℃ under N₂. After stirring for 10 hours at room temperature, the reaction mixture was quenched with saturated aq. NaHCO₃ (50 mL) . The aqueous solution was extracted with EtOAc (20 mL, 3 ×) . The combined organic phase was washed with brine (50 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (10%EtOAc in petroleum ether) to afford the desired compound 1297-2 (8.0 g, 31.7 mmol, 81.1%yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.54 (d, J = 2.8 Hz, 1H) , 8.23 (dd, J = 8.8, 2.8 Hz, 1H) , 7.86 (dt, J = 8.8, 1.2 1H) , 6.96 (t, J = 54.4 Hz, 1H) .

Step 2

To a solution of compound 1297-2 (8.0 g, 31.7 mmol) in ethanol (40 mL) and THF (80 mL) were added Zn powder (20.7 g, 317 mmol) and NH₄Cl (17.0 g, 317 mmol) at 0 ℃. After stirring for 10 hours, the reaction mixture was filtered, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (CH₂Cl₂) to afford the desired compound 1297-3 (6.50 g, 29.3 mmol, 92.2%yield) as a white solid. MS m/z (ESI) : 222.0, 224.0 [M+H] ⁺.

Step 3

Following the same procedures as in step 1 of Intermediate Example 16, compound 1297-4 was obtained (6.50 g, 18.7 mmol, 59.3%yield) . MS m/z (ESI) : 347.8, 349.8 [M+H] ⁺.

Step 4

Following the same procedures as in step 2 of Intermediate Example 6, compound 12997-5 was obtained (5.50 g, 15.8 mmol, 84.6%yield) as a white solid. MS m/z (ESI) : 347.8, 249.8 [M+H] ⁺.

Step 5

Following the same procedures as in step 2 of Intermediate Example 2, intermediate 1297-7 was obtained (20.0 mg, 0.06 mmol, 1.1%yield) . MS m/z (ESI) : 315.0.317.0 [M+H] ⁺.

Intermediate Example 22: Synthesis of intermediate 1302-2

Step 1

Following the same procedures as in step 2 of Intermediate Example 2, intermediate 1302-2 was obtained (80.0 mg, 0.26 mmol, 17.1%yield) . MS m/z (ESI) : 311.0, 313.0, 315.0 [M+H] ⁺.

Intermediate Example 23: Synthesis of intermediate 1303-3

Step 1

Following the same procedures as in step 2 of Intermediate Example 2, compound 1303-2 was obtained (300 mg, 0.96 mmol, 64.6%yield) . MS m/z (ESI) : 308.0, 310.0, 312.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 5 of Intermediate Example 6, intermediate 1303-3 was obtained (150 mg, 0.34 mmol, 28.0%yield. MS m/z (ESI) : 548.0, 550.0, 552.0 [M+H] ⁺.

Intermediate Example 24: Synthesis of intermediate 1304-6

Step 1

A solution of compound 1304-1 (5.0 g, 44.0 mmol) in SOCl₂ (75 mL) was stirred at 90 ℃for 4 days. After the reaction was completed, the reaction mixture was cooled and filtered. The filter cake was washed with EtOAc and dried to afford the desired compound 1304-2 (4.60 g, 24.0 mmol, 54.0%yield) as a white solid. MS m/z (ESI) : 189.0 [M+H] ⁺.

Step 2

To a solution of compound 1304-2 (0.50 g, 2.66 mmol) in THF (20 mL) was added LiHMDS (12.8 mL, 5.31 mmol) dropwise at –10 ℃ under N₂. After the reaction solution was stirred for 1 hour, compound 1304-2a (1.19 g, 5.31 mmol) was then added. The resulting reaction was stirred for another 2 hours and adjusted to pH = 7 using AcOH. The mixture was concentrated and diluted with aq. NaHCO₃. The aqueous solution was extracted with EtOAc (100 mL, 3 ×) . The combined organic phase was washed with brine (200 mL) , dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (25%EtOAc in petroleum ether) to afford the desired compound 1304-3 (0.83 g, 2.61 mmol, 98.0%yield) . MS m/z (ESI) : 318.0 [M+H] ⁺; ¹H NMR (400 MHz, CDCl₃) δ 12.79 (s, 1H) , 9.56 (s, 1H) , 8.39 (d, J = 7.3 Hz, 1H) , 7.99 –7.81 (m, 3H) .

Step 3

To a solution of compound 1304-3 (830 mg, 2.61 mmol) in DMA (5 mL) was added NaH (75 mg, 3.13 mmol, 60%purity) at 0 ℃. The reaction solution was heated to 200 ℃ and stirred for 18 hours. After the reaction was completed, the reaction mixture was cooled and concentrated. The residue was diluted with aq. NH₄Cl, and the resulting suspension was filtered. The filter solids were then slurred in the system of 50%EtOAc in petroleum ether to afford the desired compound 1304-4 (0.70 g, 2.34 mmol, 90.0%yield) . MS m/z (ESI) : 295.0 [M+H] ⁺.

Step 4

To a solution of compound 1304-4 (600 mg, 2.01 mmol) in POCl₃ (32 mL) was added DIPEA (779 mg, 6.03 mmol) at 0 ℃. After stirring for 0.5 hours, the reaction solution was heated to 120 ℃ and stirred for another 18 hours. After the reaction was completed, the reaction solution was cooled and concentrated. The crude was purified by silica gel column chromatography (25%EtOAc in petroleum ether) to afford the desired compound 1304-5 (600 mg, 1.89 mmol, 94.0%yield) . MS m/z (ESI) : 317.8 [M+H] ⁺.

Step 5

A solution of compound 1304-5 (300 mg, 0.95 mmol) , K₂CO₃ (392 mg, 2.84 mmol) , KI (471 mg, 2.84 mmol) , and PMB₂NH (489 mg, 1.90 mmol) in DMF (5 mL) was stirred at 120 ℃and stirred for another 18 hours. After the reaction was completed, the reaction solution was cooled and concentrated. The residue was purified by prep. HPLC to afford the desired intermediate 1304-6 (75.0 mg, 0.14 mmol, 14.0%yield) . MS m/z (ESI) : 539.0 [M+H] ⁺;

Intermediate Example 25: Synthesis of intermediate 1305-3

Step 1

To a solution of compound 1305-1 (1.0 g, 5.80 mmol) and DHP (0.98 g, 11.5 mmol) in 1, 2-dichloroethane (10 mL) was added TFA (cat. ) at room temperature. After stirring for 24 hours, the reaction mixture was diluted with CH₂Cl₂ (25 mL) . The solution was washed with 10%aq. NaHCO₃ (15 mL, 2 ×) and brine (20 mL) . The resulting organic phase was then dried over Na₂SO₄, filtered, and concentrated. The residue was purified by silica gel column chromatography (25%EtOAc in petroleum ether) to afford the desired compound 1305-2 (600 mg, 2.34 mmol, 40.3%yield) .

Step 2

Following the same procedures as in step 2 of Intermediate Example 2, intermediate 1305-3 was obtained (110 mg, 0.30 mmol, 40.0%yield) . MS m/z (ESI) : 383.3 [M+H] ⁺.

Example 1: Synthesis of compound 1226

Step 1

To a solution of intermediate 1226-6 (80 mg, 0.33 mmol) and intermediate 1226-7 (80 mg, 0.33 mmol) in DMSO (2 mL) were added N, N, N’, N’-tetramethylchloroformamidinium hexafluorophosphate (TCFH, 140 msg, 0.50 mmol) and N-methylimidazole (NMI, 135 μL, 1.69 mmol) at room temperature. After stirring for 1 hour, the reaction mixture was diluted with DMSO (1 mL) and then filtered. The filtrate was purified by prep. HPLC (Welch-Xtimate-C18-5μm-21.2*150mm. A: 10mM NH₄HCO₃; B: acetonitrile; 10-65%; retention time = 9.6 min) to afford the desired compound 1226 (5.65 mg, 0.01 mmol, 3.7%yield) as a white solid. MS m/z (ESI) : 467.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.41 (d, J = 2.0 Hz, 1H) , 8.28 –8.23 (m, 2H) , 8.15 –8.04 (m, 2H) , 7.80 (d, J = 8.4 Hz, 1H) , 7.61 (d, J = 8.4 Hz, 1H) , 7.33 (brs, 2H) , 6.62 (s, 1H) , 4.36 (s, 3H) , 4.21 –4.16 (m, 1H) , 4.12 –4.02 (m, 1H) , 3.83 (dd, J = 6.8, 9.6 Hz, 1H) , 3.59 –3.47 (m, 1H) .

Example 2: Synthesis is of compound 1235

Step 1

To a solution of intermediate 1235-4 (6 mg, 0.02 mmol) and intermediate 1235-5 (20 mg, 0.02 mmol) in 1, 4-dioxane (0.5 mL) were added Cu powder (6 mg, 0.09 mmol) , copper iodide (18 mg, 0.09 mmol) , and Cs₂CO₃ (47 mg, 0.14 mmol) at 25 ℃ under N₂. The reaction solution was heated to 95 ℃ and stirred for 16 hours. After the reaction was completed, the reaction mixture was cooled and filtered. The filter cake was washed with methanol (3 mL, 3 ×) , and the combined filtrate was concentrated. The residue was purified by prep. HPLC (Waters-Xbridge-C18-10μm-19*250mm. A: 10mM NH₄HCO₃; B: acetonitrile; 32-62%) to afford the desired compound 1235 (1.24 mg, 0.003 mmol, 15.3%yield) as a white solid. MS m/z (ESI) : 468.0 [M+H] ⁺. ¹H NMR (500 MHz, DMSO-d₆) δ: 8.63 (d, J = 2.4 Hz, 1H) , 8.23 (s, 1H) , 7.87 (dd, J = 2.4, 8.8 Hz, 1H) , 7.76 (s, 4H) , 7.59 (d, J = 8.8 Hz, 1H) , 6.89 (s, 2H) , 4.58 (d, J = 10.4 Hz, 1H) , 4.39 (s, 3H) , 4.23 (d, J = 10.4 Hz, 1H) , 1.10 –0.98 (m, 3H) , 0.30 –0.21 (m, 1H) .

Example 3: Synthesis of compound 1236

Step 1

Intermediate 1236-10 and Intermediate 1236-15 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1236-16 (60 mg, 0.08 mmol, 87.7%yield) as a white solid. MS m/z (ESI) : 755.4 [M+H] ⁺.

Step 2

A solution of compound 1236-16 (50 mg, 0.07 mmol) in TFA (0.5 mL) was stirred at 80 ℃for 3 hours. After the reaction was completed, the reaction mixture was concentrated, and the residue was purified by prep. HPLC (Waters-Xbridge-C18-10μm-19*250mm. A: 10mM NH₄HCO₃; B: acetonitrile; 35-65%) to afford the desired compound 1236 (16.9 mg, 0.03 mmol, 49.6%yield) as a white solid. MS m/z (ESI) : 514.9 [M+H] ⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.34 (s, 1H) , 8.30 (s, 1H) , 7.90 (d, J = 7.5 Hz, 1H) , 7.70 (s, 1H) , 7.55 (d, J = 8.0 Hz, 1H) , 7.46 (d, J = 1.5 Hz, 1H) , 7.38 (brs, 2H) , 4.40 (s, 3H) , 4.13 (d, J = 9.5 Hz, 1H) , 3.94 (d, J = 10.0 Hz, 1H) , 3.26 (s, 3H) , 3.00 (d, J = 17.0 Hz, 1H) , 2.81 (d, J = 17.0 Hz, 1H) .

Example 4: Synthesis of compound 1241

Step 1

Intermediate 1241-7 and Intermediate 1235-5 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1241 (2.81 mg, 0.01 mmol, 17.3%yield) as a white solid. LC-MS (ESI) : m/z = 451.9 [M+H] ⁺; ¹H NMR (500 MHz, DMSO-d₆) δ: 8.61 (d, J = 2.5 Hz, 1H) , 8.23 (s, 1H) , 7.90 (dd, J = 2.5, 9.0 Hz, 1H) , 7.74 (d, J = 8.0 Hz, 2H) , 7.59 (d, J = 9.0 Hz, 1H) , 7.55 (d, J = 8.0 Hz, 2H) , 6.89 (s, 2H) , 4.56 (t, J = 9.0 Hz, 1H) , 4.39 (s, 3H) , 4.16 (dd, J = 5.0, 10.0 Hz, 1H) , 3.86 –3.78 (m, 1H) , 1.02 –0.96 (m, 2H) , 0.89 –0.82 (m, 1H) , 0.38 –0.32 (m, 1H) .

Example 5: Synthesis of compound 1244

Step 1

Intermediate 1241-7 and Intermediate 1236-15 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1244-3 (10.0 mg, 0.01 mmol, 19.0%yield) as a yellow oil. LC-MS (ESI) : m/z = 726.0 [M+H] ⁺.

Step 2

Following the same procedures as step 2 of Example 3, compound 1244-3 was converted to compound 1244 was obtained (6.63 mg, 0.01 mmol, 97.2%yield) as a white oil. LC-MS (ESI) : m/z = 485.9 [M+H] ⁺; ¹H NMR (500 MHz, DMSO-d₆) δ: 8.27 (s, 2H) , 7.77 (d, J = 8.0 Hz, 2H) , 7.68 (s, 1H) , 7.64 (d, J = 8.0 Hz, 2H) , 7.25 (s, 2H) , 4.43 –4.33 (m, 1H) , 4.41 –3.36 (m, 4H) , 3.94 –3.83 (m, 2H) , 1.16 –1.10 (m, 1H) , 1.01 –0.93 (m, 2H) , 0.35 –0.27 (m, 1H) .

Example 6: Synthesis of compound 1246

Step 1

Compound 1236-21 and Intermediate 1226-6 were used to follow a procedure same as described in step 1 of Example 1 to give compound 1246 (4.84 mg, 0.01 mmol, 2.8%yield) as a white solid. LC-MS (ESI) : m/z = 501.3 [M+H] ⁺; ¹H NMR (500 MHz, DMSO-d₆) δ 8.34 (d, J = 8.0 Hz, 1H) , 8.26 (s, 1H) , 8.22 (s, 1H) , 8.17 (d, J = 8.0 Hz, 1H) , 8.06 (s, 1H) , 7.60 (s, 1H) , 7.34 (s, 2H) , 6.58 (s, 1H) , 4.34 (s, 3H) , 4.14 (d, J = 4.5 Hz, 1H) , 3.58 –3.45 (m, 3H) .

Example 7: Synthesis of compound 1248 and 1249

Step 1

Compound 1248 and 1249 were obtained by separation of Compound 1236 (see Example 3) via SFC purification (Column: AD, 30*250 mm, 5 μm; Flow: 100 mL/min; Eluent: A: CO₂, B: 0.1%of 7 M methanolic ammonia solution in ethanol) . Compound 1248 (5.34 mg, 0.01 mmol, 41.1%yield) was obtained as a white solid. MS m/z (ESI) : 514.9 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.32 (s, 1H) , 8.27 (s, 1H) , 7.90 (d, J = 7.6 Hz, 1H) , 7.68 (s, 1H) , 7.55 (d, J = 7.6 Hz, 1H) , 7.46 (s, 1H) , 7.26 (s, 2H) , 4.39 (s, 3H) , 4.12 (d, J = 10.0 Hz, 1H) , 3.93 (d, J = 9.6 Hz, 1H) , 3.26 (s, 3H) , 3.00 (d, J = 17.2 Hz, 1H) , 2.80 (d, J = 17.2 Hz, 1H) ; Compound 1249 (5.01 mg, 0.01 mmol, 38.5%yield) was obtained as a white solid. MS m/z (ESI) : 514.9 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.32 (s, 1H) , 8.27 (s, 1H) , 7.90 (d, J = 7.6 Hz, 1H) , 7.68 (s, 1H) , 7.55 (d, J = 8.0 Hz, 1H) , 7.46 (s, 1H) , 7.26 (s, 2H) , 4.39 (s, 3H) , 4.12 (d, J = 9.6 Hz, 1H) , 3.93 (d, J = 10.0 Hz, 1H) , 3.26 (s, 3H) , 3.00 (d, J = 16.8 Hz, 1H) , 2.80 (d, J = 16.8 Hz, 1H) .

Example 8: Synthesis of compound 1250

Step 1

Intermediate 1250-1 and Intermediate 1226-6 were used to follow a procedure same as described in step 1 of Example 1 to give compound 1250 (23.0 mg, 0.05 mmol, 17.7%yield) as a white solid. LC-MS (ESI) : m/z = 485.3 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.30 –8.25 (m, 3H) , 8.14 (d, J = 8.0 Hz, 1H) , 8.06 (s, 1H) , 7.38 (brs, 2H) , 7.31 (d, J = 12.4 Hz, 1H) , 6.59 (s, 1H) , 4.34 (s, 3H) , 4.15 (dd, J = 6.6, 11.6 Hz, 1H) , 3.81 –3.69 (m, 1H) , 3.67 –3.59 (m, 1H) , 3.57 –3.47 (m, 1H) .

Example 9: Synthesis of compound 1252

Step 1

Intermediate 1226-7 and Intermediate 1252-4 were used to follow a procedure same as described in step 1 of Example 1 to give compound 1252 (11.2 mg, 0.02 mmol, 9.4%yield) as a white solid. LC-MS (ESI) : m/z = 481.4 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 35 (d, J = 2.0 Hz, 1H) , 8.26 (s, 1H) , 8.16 (d, J = 8.0 Hz, 1H) , 8.05 (d, J = 9.6 Hz, 2H) , 7.71 –7.53 (m, 2H) , 7.21 (s, 2H) , 6.70 (s, 1H) , 4.39 (s, 3H) , 4.30 –4.16 (m, 1H) , 3.83 –3.67 (m, 1H) , 3.51 –3.42 (m, 1H) , 3.19 –3.02 (m, 1H) , 2.13 –1.98 (m, 1H) , 1.96 –1.82 (m, 1H) .

Example 10: Synthesis of compound 1262 and 1263

Step 1

Compound 1262 and 1263 were obtained by separation of Compound 1250 (see Example 8) via SFC purification (Column: ChiralPak I. D., 30*250 mm, 5 μm; Flow: 100 mL/min; Eluent: A: CO₂, B: 0.1%of 7 M methanolic ammonia solution in ethanol) . Compound 1262 (7.19 mg, 0.02 mmol, 35.8%yield) . MS m/z (ESI) : 485.1 [M+H] ⁺; ¹H NMR (400 MHz, methanol-d₄) δ 8.49 –8.45 (m, 1H) , 8.37 –8.31 (m, 1H) , 8.24 (s, 1H) , 8.11 (s, 1H) , 8.08 –8.04 (m, 1H) , 7.40 –7.34 (m, 1H) , 6.68 (s, 1H) , 4.45 (s, 3H) , 4.37 –4.31 (m, 1H) , 3.86 –3.79 (m, 1H) , 3.78 –3.71 (m, 1H) , 3.62 –3.55 (m, 1H) ; Compound 1263 (9.11 mg, 0.02 mmol, 45.5%yield) . MS m/z (ESI) : 485.1 [M+H] ⁺; ¹H NMR (400 MHz, methanol-d₄) δ 8.49 -8.43 (m, 1H) , 8.37 -8.30 (m, 1H) , 8.24 (s, 1H) , 8.11 (s, 1H) , 8.09 -8.04 (m, 1H) , 7.40 -7.33 (m, 1H) , 6.68 (s, 1H) , 4.44 (s, 3H) , 4.37 -4.31 (m, 1H) , 3.87 -3.78 (m, 1H) , 3.77 -3.70 (m, 1H) , 3.64 -3.57 (m, 1H) .

Example 11: Synthesis of compound 1264 and 1265

Step 1

Intermediate 1264-6 and Intermediate 1236-15 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1264-7 (34.0 mg, 0.05 mmol, 42.0%yield) as a white solid. MS m/z (ESI) : 725.4 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1264-7 was covered to compound 1264-9 (30.0 mg, 0.06 mmol, 98.0%yield) as a white solid. MS m/z (ESI) : 725.4 [M+H] ⁺.

Step 3

Compound 1264 and 1265 were obtained by separation of compound 1264-9 via SFC purification (Column: ChiralPak I. D., 30*250 mm, 5 μm; Flow: 100 mL/min; Eluent: A: CO₂, B: 0.2%of TFA in ethanol) . Compound 1264 (11.8 mg, 0.02 mmol, 35.8%yield) . MS m/z (ESI) : 488.2 [M+H] ⁺; ¹H NMR (400 MHz, methanol-d₄) δ 8.51 –8.44 (m, 2H) , 7.91 (s, 1H) , 7.67 (s, 1H) , 7.32 –7.28 (m, 1H) , 7.12 –7.07 (m, 1H) , 4.93 –4.89 (m, 1H) , 4.74 –4.71 (m, 1H) , 4.50 (s, 3H) , 4.17 –4.09 (m, 2H) , 3.19 –3.13 (m, 1H) , 2.99 –2.94 (m, 1H) ; Compound 1265 (11.6 mg, 0.02 mmol, 38.0%yield) . MS m/z (ESI) : 488.2 [M+H] ⁺; ¹H NMR (400 MHz, methanol-d₄) δ 8.50 –8.46 (m, 2H) , 7.92 (s, 1H) , 7.70 –7.66 (m, 1H) , 7.32 –7.28 (m, 1H) , 7.11 (s, 1H) , 4.93 –4.89 (m, 1H) , 4.74 –4.71 (m, 1H) , 4.50 (s, 3H) , 4.17 –4.09 (m, 2H) , 3.20 –3.13 (m, 1H) , 2.99 –2.94 (m, 1H) .

Example 12: Synthesis of compound 1267

Step 1

Intermediate 1267-10 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1267-6 (60.0 mg, 0.08 mmol, 87.0%yield) as a white solid. MS m/z (ESI) : 739.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1267-6 was covered to compound 1267 (11.4 mg, 0.02 mmol, 33.9%yield) as a white solid. MS m/z (ESI) : 725.4 [M+H] ⁺.

Example 13: Synthesis of compound 1268

Step 1

Intermediate 1268-8 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1268-9 (16.0 mg, 0.02 mmol, 46.1%yield) as a white solid. MS m/z (ESI) : 727.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1268-9 was converted to compound 1268 (7.0 mg, 0.01 mmol, 65.4%yield) as a white solid. MS m/z (ESI) : 487.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.81 –7.75 (m, 1H) , 7.73 –7.67 (m, 1H) , 7.56 –7.50 (m, 1H) , 7.47 –7.43 (m, 1H) , 7.41 –7.35 (m, 1H) , 6.75 (s, 2H) , 5.37 –5.28 (m, 2H) , 5.04 –4.96 (m, 2H) , 4.14 –4.08 (m, 1H) , 3.98 –3.91 (m, 1H) , 3.24 (s, 3H) , 2.94 –2.80 (m, 2H) .

Example 14: Synthesis of compound 1269 and 1270

Step 1

Compounds 1269 and 1270 were obtained by separation of Compound 1252 (see Example 9) via SFC purification (Column: AD-3, 4.6*100 mm, 3 μm; Flow: 100 mL/min; Eluent: A: CO₂, B: IPA (0.1%DEA) ) . Compound 1269 (7.63 mg, 0.01 mmol, 25.4%yield) . MS m/z (ESI) : 481.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 –8.35 (m, 1H) , 8.26 (s, 1H) , 8.17 –8.15 (m, 1H) , 8.06 –8.04 (m, 2H) , 7.66 –7.59 (m, 2H) , 7.22 (s, 2H) , 6.71 (s, 1H) , 4.39 (s, 3H) , 4.26 –4.22 (m, 1H) , 3.77 –3.73 (m, 1H) , 3.49 –3.45 (m, 1H) , 3.14 –3.09 (m, 1H) , 2.09 –1.89 (m, 2H) . Compound 1270 (7.40 mg, 0.02 mmol, 45.5%yield) . MS m/z (ESI) : 481.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 -8.35 (m, 1H) , 8.27 (s, 1H) , 8.17 -8.15 (m, 1H) , 8.06 -8.04 (m, 2H) , 7.66 –7.60 (m, 2H) , 7.23 (s, 2H) , 6.71 (s, 1H) , 4.39 (s, 3H) , 4.27 -4.20 (m, 1H) , 3.76 -3.73 (m, 1H) , 3.49 –3.45 (m, 1H) , 3.14 -3.11 (m, 1H) , 2.08 –1.91 (m, 2H) .

Example 15: Synthesis of compound 1271

Step 1

A solution of compound 1236 (see Example 3) (20.0 mg, 0.04 mmol) and Pd on charcoal (5.0 mg, 10%purity) in methanol (1 mL) was degassed by purging hydrogen gases for 3 times at room temperature. The reaction mixture was stirred under H₂ for 10 hours. After the reaction was completed, the reaction mixture was filtered. The resulting filtrate was concentrated to give a residue. The residue was further purified by prep. HPLC (Waters-Xbridge-C18-10μm-19*250mm. A: 10mM NH₄HCO₃; B: acetonitrile; 10-95%) to afford the desired compound 1271 (1.5 mg, 0.01 mmol, 8.0%yield) . MS m/z (ESI) : 481.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.42 (s, 1H) , 8.23 (s, 1H) , 7.99 –7.91 (m, 1H) , 7.80 (d, J = 7.80 Hz, 1H) , 7.60 (d, J = 9.2 Hz, 1H) , 7.50 (d, J = 7.6 Hz, 1H) , 7.45 (s, 1H) , 6.91 (s, 2H) , 4.37 (s, 3H) , 4.31 –4.22 (m, 2H) , 3.24 (s, 3H) , 3.05 (d, J = 17.2 Hz, 1H) , 2.92 (d, J = 17.2 Hz, 1H) .

Example 16: Synthesis of compound 1275

Step 1

Intermediate 1236-15 and Intermediate 1275-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1275-11 (30.0 mg, 0.04 mmol, 36.0%yield) . MS m/z (ESI) : 742.0 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1275-11 was converted to compound 1275 (20.0 mg, 0.04 mmol, 98.0%yield) . MS m/z (ESI) : 502.2 [M+H] ⁺; ¹H NMR (400 MHz, Methanol-d₄) δ 8.35 (s, 1H) , 8.21 (s, 1H) , 7.97 –7.93 (m, 1H) , 7.76 (s, 1H) , 7.68 –7.63 (m, 1H) , 7.44 (s, 1H) , 4.92 (s, 2H) , 4.44 (s, 3H) , 4.31 –4.25 (m, 1H) , 4.11 –4.05 (m, 2H) , 3.98 –3.93 (m, 1H) , 3.19 –3.12 (m, 1H) , 2.79 –2.72 (m, 1H) .

Example 17: Synthesis of compound 1277

Step 1

Intermediate 1277-5 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1277-7 (3.0 mg, 0.01 mmol, 21.4%yield) . MS m/z (ESI) : 709.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1277-7 was converted to compound 1277 (0.85 mg, 0.01 mmol, 6.4%yield) . MS m/z (ESI) : 469.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.08 –8.04 (m, 1H) , 7.80 (d, J = 8.0 Hz, 1H) , 7.59 (d, J = 9.2 Hz, 1H) , 7.53 –7.48 (m, 2H) , 7.44 (s, 1H) , 6.45 (s, 2H) , 5.30 –5.25 (m, 2H) , 5.06 –4.90 (m, 2H) , 3.23 (s, 3H) , 3.05 –3.02 (m, 2H) , 2.90 –2.86 (m, 2H) .

Example 18: Synthesis of compound 1281

Step 1

Intermediate 1268-11 and Intermediate 1281-3 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1281 (3.0 mg, 0.01 mmol, 18.2%yield) . MS m/z (ESI) : 513.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.78 –7.76 (m, 1H) , 7.70 –7.67 (m, 1H) , 7.55 –7.52 (m, 1H) , 7.39 –7.36 (m, 2H) , 6.74 (s, 2H) , 5.31 –5.29 (m, 2H) , 5.01 –4.99 (m, 2H) , 4.09 –4.06 (m, 1H) , 3.94 –3.92 (m, 1H) , 2.91 –2.77 (m, 3H) , 1.07 –1.04 (m, 2H) , 0.89 –0.86 (m. 2H) .

Example 19: Synthesis of compound 1282

Step 1

Compound 1282-1 and Intermediate 1236-10 were used to following the same procedures as in step 1 of Example 2 to give compound 1282 (9.06 mg, 0.02 mmol, 14.5%yield) . MS m/z (ESI) : 441.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 713.89 (s, 1H) , 8.47 (s, 2H) , 8.24 (s, 1H) , 8.15 –8.06 (m, 2H) , 7.81 (d, J = 7.6 Hz, 1H) , 7.73 (d, J = 9.2 Hz, 1H) , 7.50 (d, J = 7.6 Hz, 1H) , 7.45 (d, J = 1.6 Hz, 1H) , 4.51 –3.95 (m, 2H) , 3.23 (s, 3H) , 3.10 (d, J = 16.8 Hz, 1H) , 2.90 (d, J = 16.8 Hz, 1H) , 2.31 (d, J = 1.2 Hz, 3H) .

Example 20: Synthesis of compound 1285

Step 1

Intermediate 1285-4 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1285 (1.01 mg, 0.01 mmol, 0.6%yield) . MS m/z (ESI) : 503.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.85 (d, J = 7.6 Hz, 1H) , 7.73 (s, 1H) , 7.69 (s, 1H) , 7.54 (d, J = 7.6 Hz, 1H) , 7.45 (s, 1H) , 6.83 (s, 2H) , 5.32 (d, J = 4.0 Hz, 2H) , 5.01 (s, 2H) , 4.06 (d, J = 9.6 Hz, 1H) , 3.85 (d, J = 9.6 Hz, 1H) , 3.25 (s, 3H) , 2.94 (d, J = 17.2 Hz, 1H) , 2.78 (d, J = 17.2 Hz, 1H) .

Example 21: Synthesis of compound 1286

Step 1

Intermediate 1226-6 and Intermediate 1286-4 were used to follow a procedure same as described in step 1 of Example 1 to give compound 1286 (2.01 mg, 0.01 mmol, 2.0%yield) . LC-MS (ESI) : m/z = 455.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.19 (s, 1H) , 8.11 –8.02 (m, 2H) , 7.77 –7.70 (m, 2H) , 7.57 (d, J = 8.8 Hz, 1H) , 6.82 –6.72 (m, 2H) , 6.59 (s, 1H) , 5.34 –5.24 (m, 2H) , 5.02 –4.96 (m, 2H) , 4.20 –4.12 (m, 1H) , 4.08 –3.95 (m, 1H) , 3.72 –3.65 (m, 1H) , 3.54 –3.45 (m, 1H) .

Example 22: Synthesis of compound 1289

Step 1

Intermediate 1226-6 and Intermediate 1289-5 were used to follow a procedure same as described in step 1 of Example 1 to give compound 1289 (2.0 mg, 0.004 mmol, 2.0%yield) as a white solid. LC-MS (ESI) : m/z = 473.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.26 –8.24 (m, 1H) , 8.14 –8.12 (m, 1H) , 8.06 (s, 1H) , 7.70 –7.68 (m, 1H) , 7.31 –7.28 (m, 1H) , 6.90 (s, 2H) , 6.56 (s, 1H) , 5.32 –5.26 (m, 2H) , 4.98 –4.97 (m, 2H) , 4.14 –4.11 (m, 2H) , 3.72 –3.70 (m, 1H) , 3.53 –3.51 (m, 1H) .

Example 23: Synthesis of compound 1290

Step 1

Intermediate 1267-10 and Intermediate 1290-3 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1290-9 (2.0 mg, 0.003 mmol, 3.6%yield) as a white solid. MS m/z (ESI) : 712.4 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1290 was obtained (5.0 mg, 0.01 mmol, 75.5%yield) as a white solid. MS m/z (ESI) : 472.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.27 –8.25 (m, 2H) , 7.89 -7.87 (m, 1H) , 7.81 –7.79 (m, 2H) , 7.39 -7.36 (m, 1H) , 7.19 (s, 2H) , 5.17 (s, 2H) , 4.36 –4.33 (m, 4H) , 4.04 -4.02 (m, 1H) , 3.30 –3.19 (m, 1H) , 2.84 -2.80 (m, 1H) .

Example 24: Synthesis of compound 1291 and 1292

Step 1

Compounds 1291 and 1292 were obtained by separation of Compound 1285 (see Example 20) via SFC purification (Waters 150 preparative SFC; Flow: 100 mL/min; Eluent: A: CO₂, B: Methanol (0.1%ammonia, 7 M in methanol) ) . Compound 1291 (15.4 mg, 0.03 mmol, 39.0%yield) . MS m/z (ESI) : 503.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.51 –8.44 (m, 2H) , 7.91 (s, 1H) , 7.67 (s, 1H) , 7.32 –7.28 (m, 1H) , 7.12 –7.07 (m, 1H) , 4.93 –4.89 (m, 1H) , 4.74 –4.71 (m, 1H) , 4.50 (s, 3H) , 4.17 –4.09 (m, 2H) , 3.19 –3.13 (m, 1H) , 2.99 –2.94 (m, 1H) ; Compound 1292 (11.8 mg, 0.03 mmol, 39.0%yield) . MS m/z (ESI) : 503.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.88 –7.83 (m, 1H) , 7.73 (s, 1H) , 7.69 (s, 1H) , 7.56 –7.51 (m, 1H) , 7.47 –7.44 (m, 1H) , 6.83 (s, 2H) , 5.32 (t, J = 3.5 Hz, 2H) , 5.04 –4.97 (m, 2H) , 4.09 –4.04 (m, 1H) , 3.88 –3.83 (m, 1H) , 3.25 (s, 3H) , 2.94 (d, J = 17.0 Hz, 1H) , 2.78 (d, J =17.0 Hz, 1H) .

Example 25: Synthesis of compound 1293 and 1294

Step 1

Compounds 1293 and 1294 were obtained by separation of Compound 1281 (see Example 18) via SFC purification (Column: AD-3, 4.6*100 mm, 3 μm; Flow: 100 mL/min; Eluent: A: CO₂, B: Methanol (0.1%DEA) ) . Compound 1293 (4.12 mg, 0.007 mmol, 35.2%yield) . MS m/z (ESI) : 513.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.78 –7.76 (m, 1H) , 7.70 –7.67 (m, 1H) , 7.55 –7.53 (m, 1H) , 7.39 –7.36 (m, 2H) , 6.74 (s, 2H) , 5.30 –5.29 (m, 2H) , 5.00 –4.49 (m, 2H) , 4.09 –4.06 (m, 1H) , 3.94 –3.92 (m, 1H) , 2.91 –2.77 (m, 3H) , 1.05 –1.04 (m, 2H) , 0.87 –0.86 (m, 2H) ; Compound 1294 (1.90 mg, 0.003 mmol, 16.2%yield) . MS m/z (ESI) : 513.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.77 -7.76 (m, 1H) , 7.70 -7.67 (m, 1H) , 7.55 -7.53 (m, 1H) , 7.39 -7.36 (m, 2H) , 6.74 (s, 2H) , 5.31 -5.29 (m, 2H) , 5.01 -4.99 (m, 2H) , 4.09 -4.06 (m, 1H) , 3.94 -3.92 (m, 1H) , 2.91 -2.77 (m, 3H) , 1.07 -1.04 (m, 2H) , 0.89 -0.86 (m, 2H) .

Example 26: Synthesis of compound 1295

Step 1

Intermediate 1295-5 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1295-6 (80.0 mg, 0.11 mmol, 78.0%yield) . MS m/z (ESI) : 723.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1295-6 was converted to compound 1295 (10.0 mg, 0.02 mmol, 18.0%yield) . MS m/z (ESI) : 483.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.91 –7.86 (m, 1H) , 7.56 –7.52 (m, 1H) , 7.49 –7.45 (m, 2H) , 7.44 –7.41 (m, 1H) , 6.53 (s, 2H) , 5.30 (t, J = 3.6 Hz, 2H) , 5.00 (t, J = 3.6 Hz, 2H) , 4.05 (d, J = 10.0 Hz, 1H) , 3.90 (d, J = 10.0 Hz, 1H) , 3.24 (s, 3H) , 2.93 (s, 1H) , 2.83 (s, 1H) , 2.41 (s, 3H) .

Example 27: Synthesis of compound 1296

Step 1

Intermediate 1285-4 was used to follow a procedure same as in step 5 of Intermediate Example 6 to give compound 1296-1.

Step 2

Compound 1296-1 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1296-2 (200 mg, 0.27 mmol, 38.0%yield) . MS m/z (ESI) : 743.2 [M+H] ⁺.

Step 3

A solution of compound 1296-2 (100 mg, 0.13 mmol) , Pd [Pd (tBu) ₃] ₂ (27.0 mg, 0.05 mmol) , and Zn (CN) ₂ (47.0 mg, 0.40 mmol) in NMP was stirred at 180 ℃ in a microwave reactor for 20 minutes. After the reaction was completed, the reaction mixture was cooled and filtered. The filter cake was washed with EtOAc and water, and dried to afford the compound 1296-3 (90.0 mg, 0.12 mmol, 91.0%yield) . MS m/z (ESI) : 734.2 [M+H] ⁺.

Step 4

Following the same procedures as in step 2 of Example 3, compound 1296-3 was converted to compound 1296 (2.85 mg, 0.01 mmol, 4.0%yield) . MS m/z (ESI) : 494.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.06 (s, 1H) , 7.87 (d, J = 7.6 Hz, 1H) , 7.82 (s, 1H) , 7.53 –7.45 (m, 2H) , 6.97 (s, 2H) , 5.33 (d, J = 3.6 Hz, 2H) , 5.04 (d, J = 3.6 Hz, 2H) , 4.27 (d, J = 9.6 Hz, 1H) , 3.98 (d, J = 9.6 Hz, 1H) , 3.26 (s, 3H) , 2.99 (d, J = 17.2 Hz, 1H) , 2.82 (d, J = 17.2 Hz, 1H) .

Example 28: Synthesis of compound 1297

Step 1

Intermediate 1297-7 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1297 (0.97 mg, 0.01 mmol, 50.0%yield) . MS m/z (ESI) : 519.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 (s, 1H) , 8.00 (s, 1H) , 7.82 (d, J = 7.6 Hz, 1H) , 7.55 (d, J = 7.6 Hz, 1H) , 7.47 (s, 1H) , 7.23 (t, J = 54.8 Hz, 2H) , 7.05 (s, 2H) , 5.06 (s, 2H) , 4.94 (s, 2H) , 4.21 –4.01 (m, 2H) , 3.26 (s, 3H) , 2.96 (d, J = 6.0 Hz, 2H) .

Example 29: Synthesis of compound 1298

Step 1

Intermediate 1285-4 and Intermediate 1275-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1298 (1.56 mg, 0.001 mmol, 2.9%yield) . MS m/z (ESI) : 490.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.06 (s, 1H) , 7.87 (d, J = 7.6 Hz, 1H) , 7.82 (s, 1H) , 7.53 –7.45 (m, 2H) , 6.97 (s, 2H) , 5.33 (d, J = 3.2 Hz, 2H) , 5.04 (d, J = 3.5 Hz, 2H) , 4.27 (d, J = 9.6 Hz, 1H) , 3.98 (d, J = 9.6 Hz, 1H) , 3.26 (s, 3H) , 2.99 (d, J = 17.2 Hz, 1H) , 2.82 (d, J = 17.2 Hz, 1H) .

Example 30: Synthesis of compound 1299

Step 1

Intermediate 1268-11 and Intermediate 1275-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1299 (4.85 mg, 0.01 mmol, 9.1%yield) . MS m/z (ESI) : 474.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.94 –7.90 (m, 1H) , 7.70 –7.63 (m, 2H) , 7.54 –7.50 (m, 1H) , 7.38 –7.33 (m, 1H) , 6.72 (s, 2H) , 5.33 –5.26 (m, 2H) , 5.02 –4.96 (m, 2H) , 4.90 –4.85 (m, 2H) , 4.08 –4.02 (m, 1H) , 3.99 –3.94 (m, 1H) , 3.92 –3.80 (m, 2H) , 3.05 –2.96 (m, 1H) , 2.68 –2.61 (m, 1H) .

Example 31: Synthesis of compound 1300

Step 1

Intermediate 1277-5 and Intermediate 1275-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1300-5 (120 mg, 0.17 mmol, 88.0%yield) . MS m/z (ESI) : 695.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1300-5 was converted to compound 1300 (134 mg, 0.07 mmol, 43.0%yield) . MS m/z (ESI) : 456.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 7.94 –7.90 (m, 1H) , 7.70 –7.63 (m, 2H) , 7.54 –7.50 (m, 1H) , 7.38 –7.33 (m, 1H) , 6.72 (s, 2H) , 5.33 –5.26 (m, 2H) , 5.02 –4.96 (m, 2H) , 4.90 –4.85 (m, 2H) , 4.08 –4.02 (m, 1H) , 3.99 –3.94 (m, 1H) , 3.92 –3.80 (m, 2H) , 3.05 –2.96 (m, 1H) , 2.68 –2.61 (m, 1H) .

Example 32: Synthesis of compound 1302

Step 1

Intermediate 1302-2 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1302 (9.21 mg, 0.02 mmol, 9.3%yield) . MS m/z (ESI) : 515.3, 517.3 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.51 (s, 1H) , 8.29 (s, 1H) , 7.88 (d, J = 7.6 Hz, 1H) , 7.68 (s, 1H) , 7.55 (d, J = 7.6 Hz, 1H) , 7.47 (d, J = 1.6 Hz, 1H) , 7.05 (s, 2H) , 4.36 (s, 3H) , 4.12 (d, J = 9.6 Hz, 1H) , 3.90 –3.87 (m, 1H) , 3.26 (s, 3H) , 2.98 (d, J = 17.0 Hz, 1H) , 2.79 (d, J = 17.0 Hz, 1H) .

Example 33: Synthesis of compound 1303

Step 1

Intermediate 1303-3 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1303-4 (65.0 mg, 0.09 mmol, 34.6%yield) . MS m/z (ESI) : 752.2, 754.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1303-4 was converted to compound 1303 (12.5 mg, 0.02 mmol, 28.2%yield) . MS m/z (ESI) : 512.1, 514.1 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 10.08 (s, 1H) , 8.91 –8.84 (m, 2H) , 8.25 (d, J = 5.6 Hz, 1H) , 7.95 (d, J = 7.6 Hz, 1H) , 7.66 (m, 3H) , 7.57 (d, J = 7.6 Hz, 1H) , 7.47 (d, J = 1.6 Hz, 1H) , 4.22 (d, J = 9.6 Hz, 1H) , 3.90 (d, J = 9.6 Hz, 1H) , 3.28 (s, 3H) , 3.02 (d, J = 17.0 Hz, 1H) , 2.78 (d, J = 17.0 Hz, 1H) .

Example 34: Synthesis of compound 1304

Step 1

Intermediate 1304-6 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1303-7 (50.0 mg, 0.05 mmol, 44.0%yield) . MS m/z (ESI) : 741.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1304-7 was converted to compound 1304 (8.0 mg, 0.02 mmol, 23.0%yield) . MS m/z (ESI) : 501.0 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 8.44 (s, 1H) , 7.90 –7.83 (m, 1H) , 7.64 –7.51 (m, 4H) , 7.47 (s, 1H) , 4.19 –4.13 (m, 1H) , 3.89 –3.82 (m, 1H) , 3.27 (s, 3H) , 3.03 –2.96 (m, 1H) , 2.78 (s, 1H) .

Example 35: Synthesis of compound 1305

Step 1

Intermediate 1305-3 and Intermediate 1236-10 were used to follow a procedure same as described in step 1 of Example 2 to give compound 1305-4 (120 mg, 0.10 mmol, 58.0%yield) . MS m/z (ESI) : 585.2 [M+H] ⁺.

Step 2

Following the same procedures as in step 2 of Example 3, compound 1305-4 was converted to compound 1305 (8.0 mg, 0.02 mmol, 15.0%yield) . MS m/z (ESI) : 501.2 [M+H] ⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 15.00 (s, 1H) , 9.04 (s, 1H) , 8.38 (s, 1H) , 7.89 –7.83 (m, 2H) , 7.58 –7.53 (m, 1H) , 7.50 –7.46 (m, 1H) , 4.14 –4.09 (m, 1H) , 3.96 –3.91 (m, 1H) , 3.26 (s, 3H) , 3.01 –2.95 (m, 1H) , 2.88 –2.82 (m, 1H) .

Biological examples:

Example A. PRMT5 inhibition based on TR-FRET assay

Example compounds were prepared in the form of stock solutions in DMSO at 10mM and 20 mM. The stock solutions were further formulated to serial diluted solutions for testing, wherein 4 μL of each sample of the test compounds was introduced into a 384-well microplate. Then, 5 μL of a mixture solution of PRMT5 enzyme and MTA was added to each well of said microplate, and the test samples were incubated for 30 minutes at room temperature.

Histone H4 peptide and S-adenosylmethionine were mixed to produce a PRMT5 substrate solution and into each well of the microplate was added 8 μL of the PRMT5 substrate solution to initiate the reaction. The reaction samples were incubated for 90 minutes at room temperature and then each well received a mixed solution of Protin-A-Eu, anti-Histone H4 (symmetric dimethyl R3) antibody and streptavidin-d2 to terminate the reaction. The microplate was placed on a BMG reader for reading at wavelengths of 615 nm and 665nm in accordance with TR-FRET protocol. Percentage inhibition and concentration of the test compounds were plotted using GraphPad Prism to calculate the IC₅₀.

Table 1: TR-FRET MTA+ activity data

Example B. Cell proliferation assay in MTAPdel HCT116 and Wild Type HCT116 cell lines

On day 0, HCT116 MTAPdel cell and HCT116 wild type cell lines were seeded into a 384-well microplate supplemented with McCOYS 5A containing 10%fetal bovine serum and pen/strep and incubated overnight in an incubator at 37℃, 5%CO₂. On the following day, 40μL of DMSO vehicle, as control, and different concentrations of Example compounds were added to independent wells of the microplate. The microplate was placed in the incubator at 37℃, 5%CO₂ for incubation for ten days.

On Day 10, into each well of the microplate was added 40μL CTG solution (Promega, Cat No. G7573) and the microplate was placed in the incubator at 37℃, 5%CO₂ for incubation for 30 minutes. The inhibitory effect was evaluated using a CTG assay kit (CellTiterGlo; Promega cat. no G7573) in accordance with the manufacturer’s instructions. IC₅₀ values for each compound after 10 days of treatment were calculated using GraphPad PRISM software.

Cauculation of IC₅₀:

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

X: log of inhibitor concentration

Y: %Inhibition.

Table 2 shows cell amplification activity in MTAPdel HCT116 and Wild Type HCT116 cell lines (10 days) .

Example C: In vivo pharmacokinetics studies

Formulation preparation:

Compounds to be tested were formulated into solutions for intravenous (IV) administration or suspensions for oral (PO) administration, at different concentrations. The IV formulations were prepared with 20%SBE-β-CD in 50 mM citric acid, pH 5 solution, and the PO formulations were prepared with water containing 0.5%methylcellulose (viscosity 4,000 cP) and 0.2%Tween 80 (polyoxyetbylenesorbitan monooleate) .

Dosage administration and sample collection:

The formulations were administered intravenously via tail vein at dose of 3 mg/Kg or orally via oral gavage at dose of 10mg/kg to male CD-I mice, 6-8 weeks. Blood was collected via facial vein bleeding. Blood samples were put on ice and centrifuged to obtain plasma sample (2000g, 5 min at 4 ℃) within 15 minutes of collection. The concentration of tested compounds in mouse plasma was determined by LC/MS/MS.

The pharmacokinetic parameters were calculated using Phoenix WinNonlin V 8.2 statistics software with a non-compartmental model. Key parameters such as half-life (T_1/2) from IV dosing and maximum concentration (C_max) , AUC_0-t, oral bioavailability (F%) from PO dosing were obtained, summarized as follows.

Table 3: Pharmacokinetic (PK) Parameters of Exemplary Compounds

Pharmacokinetic studies showed desired PK parameters that provide drug exposure and flexibility for treatment of cancer.

Example D: In vivo pharmacodynamic evaluation of human lung cancer LU99 cell subcutaneous xenograft tumor BALB/c nude mouse model

The BALB/c nude mouse model of human lung cancer LU99 xenograft tumors with homozygous MTAP gene deletion was used to determine the inhibitory effect of compounds in MTAP gene deleted cancer.

Cell culture: LU99 cells were maintained in vitro as monolayer culture in RPMI-1640 Medium supplemented with 10%fetal bovine serum at 37℃ in an atmosphere of 5%CO₂ in air. The tumor cells were routinely subcultured weekly by trypsin-EDTA treatment, not to exceed 4-5 passages. The cells growing in an exponential growth phase were harvested and counted for tumor inoculation.

Method for Tumor Inoculation and Randomization: 0.1 mL of LU99 tumor cells (1 × 10⁷) was subcutaneously inoculated on the right flank of each mouse. When the average tumor volume reached 150 mm³, they were divided into groups by randomization to minimize group to group variation in tumor volume.

Compounds were suspended in an aqueous solution containing 0.5% (w/v) methyl cellulose (MC) and 0.2% (v/v) Tween 80, and administered at 10mL/kg Volumetric gavage was given to mice once a day (QD) .

The measurement of tumor size was conducted twice per week with a caliper and the tumor volume (mm³) was estimated using the formula TV = a × b²/2, where “a” and “b” are the long and short diameters of a tumor, respectively. The TVs are used for calculation of the tumor growth inhibition rate (TGI (%) , an indicator of antitumor effectiveness) using the formula: TGI (%) = (1-T/C) × 100%, where “T” and “C” are the mean relative volumes (%tumor growth) of the tumors in the treated and the control groups, respectively.

sDMA levels in tumor from treated and control groups were also determined to evaluate the PRMT5 inhibition in tumors. Tumors from each mouse were weighted and homogenized in RIPA buffer supplemented with protease inhibitor (Halt Protease&Phosphatase inhibitor cocktail, EDTA-free (100x) , ThermoFisher Scientific, Catalog #: 78445) . Lysate were centrifuged at 12,000 rpm for 15 min at 4 ℃ to remove debris. Total protein concentrations of lysate were determined by BCA assay (ThermoFisher Scientific, Catalog #: 23227) . Equal amount of total proteins from each tumor were separated on SDS-PAGE gel, and sDMA levels were determined by Western Blotting.

Western Blotting: Total 20μg protein from each sample were loaded for electrophoresis and the gels run at constant voltage of 120 min in MOPS running buffer. After transferring to a nitrocellulose membrane using Transblot (0.3A, 1.5h) , the membrane was blocked with blocking buffer for 1 hour at room temperature. The membrane was then incubated overnight at 4℃ with primary antibody (SYM11, Sigma, Catalog #: 07-413) . The membrane was washed 6 times with 1 x 0.5%TBST and incubated with secondary antibody (anti-rabbit or anti-mouse IgG, Li-Cor 926-32211 or 926-68070) for 1 hour at room temperature. The membrane was washed 6 times again with 1 x 0.5%TBST and the result was detected with Odyssey CLx imager. The scale of the target band was analyzed using Image studio. Normalization with control was applied to calculate the related expression of groups following the equations as below: Related expression = Densitytarget /Densitycontrol

The result showed that Compound 263 and Compound 292, administered QD at doses of 25 mg/kg, demonstrated significant anti-tumor effect. After 21 days of administration, the tumor growth inhibition rate (TGI%) was 95.6%, and 94.4%, respectively. The sDMA level was measured from tumor samples collected at the time around C_max (T_max) after final dosing. Comparing to control, significant sDMA level reductions (>90%) were observed for Compound 1263 and Compound 1292.

While the invention has been described and illustrated with reference to specific embodiments thereof, these descriptions and illustrations are not limiting. It will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention as defined by the appended claims. Illustrations may not necessarily be drawn to scale. Differences may exist between the process reproductions in this disclosure and actual equipment due to manufacturing processes and tolerances. There may be other embodiments of the invention not specifically illustrated. The specification and drawings are to be regarded as illustrative rather than restrictive. Modifications may be made to adapt a particular situation, material, composition of matter, method or process to the object, spirit and scope of the invention. All such modifications are intended to be within the scope of the appended claims. Although the methods disclosed herein have been described with reference to specific operations performed in a specific order, it should be understood that these operations may be combined, subdivided, or reordered to form equivalent methods without departing from the teachings of the invention. Accordingly, the order and grouping of operations are not limitations of the invention unless specifically indicated herein.

Claims

A compound of formula (I) or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled:

wherein:

X₁, X₂ and X₃ are each independently selected from CR and N; preferably X₁, X₂ and X₃ are each independently selected from CR;

ring A, which is fused withis selected from a 5-6 membered cycloalkyl, a 5-6 membered heterocyclyl, a 5-6 membered aryl and a 5-6 membered heteroaryl, or is absent, and preferably ring A is selected from a 5-membered heterocyclyl and a 5-6 membered heteroaryl or is absent, and more preferably ring A is 1, 2-oxadiazole, 1, 2-thiazole, tetrahydrofuran or pyridine; whereindenotes the position fused with ring A;

ring B is a 5-6 membered heterocyclic group, and preferably the heteroatom of ring B is connected to L;

ring C is selected from a 5-6 membered heterocyclyl, a 5-6 membered aryl, a 5-6 membered heteroaryl, a 8-11 membered fused bicyclic heterocyclyl, a 8-11 membered fused bicyclic aryl and a 8-11 membered fused bicyclic heteroaryl;

L is -C (=O) -;

idenotes the connection between ring B and ring C; the connection is selected from: (1) a direct bond; (2) ring B and ring C being fused to form a fused tricyclic ring; and (3) ring B and ring C forming a spiro ring;

each R is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably hydrogen, deuterium, halogen or C₁-C₆ alkyl;

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, oxo (=O) , cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably hydrogen, deuterium, amino or C₁-C₆ alkyl;

each R_B is independently hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, amino, C₁-C₆ alkyl, C₃-C₆ cycloalkyl-C₁-C₆ alkyl-, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , or

two R_B attached to the same carbon atom together with the carbon atom to which they are attached form a C₃-C₈ cycloalkyl group, or

R_B attached to different carbon atoms together with the carbon atoms to which they are attached form a C₃-C₇ cycloalkyl group;

each R_C is independently hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, mercapto, amino, C₁-C₆ alkyl optionally substituted with 1-3 R_Z, methylene optionally substituted with 1-3 R_Z, C₂-C₆ alkenyl optionally substituted with 1-3 R_Z, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ alkylthio, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -NH- (C₁-C₆ haloalkyl) , -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , C₁-C₆ haloalkoxy, C₁-C₆ haloalkylthio, pentafluorosulfur, -S (O) ₂- (C₁-C₆ alkyl) , -S (=O) (C₁-C₆ alkyl) (=N) R_Z, -P (=O) (C₁-C₆ alkyl) ₂, C₂-C₆ haloalkenyl, 3-6 membered cycloalkyl optionally substituted by 1-3 R_Z, 5-8 membered bridged cycloalkyl optionally substituted by 1-3 R_Z, 4-7 membered heterocyclyl optionally substituted by 1-3 R_Z, 5-6 membered aryl optionally substituted by 1-3 R_Z, 5-6 membered heteroaryl optionally substituted by 1-3 R_Z, or

two R_C connected to the same carbon atom together with the carbon atom to which they are connected form a C₃-C₈ cycloalkyl, or

R_C connected to different carbon atoms together with the carbon atom to which they are connected form a C₃-C₇ cycloalkyl or a 3-7 membered heterocyclyl, or

two R_C connected to the same carbon atom together with the carbon atom to which they are connected form C=T, wherein T is selected from CR_ER_F, and R_E and R_F are each independently selected from hydrogen, deuterium, halogen and C₁-C₆ alkyl;

each R_Z is independently selected from hydrogen, deuterium, halogen, hydroxyl, oxo, cyano, amino, pentafluorosulfur, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , 3-6 membered cycloalkyl, -NH- (C₁-C₆ haloalkyl) and -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) ;

m is 0 or 1; wherein when m is 0, it means that L is a direct bond; when m is 1, L is -C (=O) -, and ring B and ring C are fused to form a fused tricyclic ring;

n is 0, 1, 2, 3 or 4;

p is 0, 1, 2, 3 or 4; and

q is 0, 1, 2, 3 or 4.
The compound according to Claim 1, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein the compound has formula (I-1) :

wherein ring A, ring B, ring C, L, m, R_A, R_B, R_C, n, p and q are as defined above; preferably:

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl , - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl.
The compound according to Claim 1 or 2, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein the compound has formula (IA-1) :

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; and ring B and ring C form a spiro ring.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein the compound has formula (IA-1-1) :

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; and ring B and ring C form a spiro ring; preferably:

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein ring B is a 5-6 membered heterocyclic group containing at least one N; preferably:

is selected from more preferably

wherein *denotes the spiral atom; and

R_B and p are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein:

p is at least 1, and at least one R_B is an oxo group; preferably, ring B is a 5-6 membered heterocyclic group containing at least one N, p is at least 1, and at least one R_B is an oxo group; more preferably:

is selected from

wherein *denotes the spiral atom.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: ring C is selected from a fused bicyclic ring formed by a 5-membered ring fused to a 6-membered ting and a fused bicyclic ring formed by a 6-membered ring fused to a 6-membered ring; preferably, ring C is selected from a fused bicyclic ring formed by: a 5-membered heterocyclyl fused to a 6-membered aryl, a 5-membered heterocyclyl fused to 6-membered heteroaryl, a 6-membered heterocyclyl fused to 6-membered aryl, and a 6-membered heterocyclyl fused to 6-membered heteroaryl; more preferably

is selected from

wherein *denotes the spiral atom;

q1 is 0, 1, 2 or 3; and

R_C and q are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein:

is selected from

wherein *represents the spiral atom.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IA-1-2) ,

wherein:

Z₁ is selected from CH₂, NH and O, wherein H is optionally replaced by R_C;

Z₂, Z₃, Z₄ and Z₅ are each independently selected from CH and N, wherein H is optionally replaced by R_C;

m₁ and m₂ are each independently 0 or 1, preferably when one of m₁ and m₂ is 0, then the other is 1;

p1 is 0, 1, 2 or 3; and

ring A, X₁, X₂, X₃, R_A, R_B, R_C, n and q are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IA-1-2-1) , formula (IA-1-2-2) , formula (IA-1-2-3) , formula (IA-1-2-4) or formula (IA-1-2-5) :
preferably
preferably
preferably
preferably
preferably

wherein ring A, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above;

p1 and q1 are independently 0, 1, 2 or 3; and

Z₂, Z₃, Z₄ and Z₅ are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IB-1) :

wherein ring A, ring B, ring C, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein:

the compound has formula (IB-1-1) :

wherein ring A, ring B, ring C, R_A, R_B, R_C, n, p and q are as defined above, preferably

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: ring B is a 5-6 membered heterocyclic group containing at least one N; preferably:

is selected from

more preferably, #denotes the connection with (as defined above) , $ denotes the connection with ring C; and

R_B and p are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: p is at least 1, and at least one R_B is an oxo group; preferably, ring B is a 5-6 membered heterocyclic group containing at least one N, p is at least 1, and at least one R_B is an oxo group; more preferably:

is selected from

wherein #denotes the connection with (as defined above) , and $ denotes the connection with ring C.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein:

is selected from and

R_c and q are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: is selected from
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IB-1-2) :

whereindenotes a single bond or a double bond;

p1 is 0, 1, 2 or 3; and

Rring A, X₁, X₂, X₃, R_A, R_B, R_C, n and q are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein:

the compound has formula (IC-1) :

wherein ring A, ring B, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; ring B and ring C are fused to form a fused tricyclic ring; and

ring C is selected from 8-11 membered fused bicyclic ring, preferably 8-11 membered fused bicyclic heterocyclic group, 8-11 membered fused bicyclic aryl group or 8-11 membered fused bicyclic heteroaryl group.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IC-1-1) :

wherein ring A, ring B, R_A, R_B, R_C, n, p and q are as defined above; preferably

each R_A is independently hydrogen, deuterium, halogen, hydroxyl, cyano, amino, C₁-C₆ alkyl, - (C₁-C₆ alkyl) -OH, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, -NH (C₁-C₆ alkyl) , -N (C₁-C₆ alkyl) (C₁-C₆ alkyl) , -O- (C₁-C₆ haloalkyl) , -NH- (C₁-C₆ haloalkyl) or -N (C₁-C₆ haloalkyl) (C₁-C₆ haloalkyl) , preferably deuterium, amino or C₁-C₆ alkyl;

ring B and ring C are fused to form a fused tricyclic ring; and

ring C is is selected from 8-11 membered fused bicyclic ring, preferably 8-11 membered fused bicyclic heterocyclic group, 8-11 membered fused bicyclic aryl group or 8-11 membered fused bicyclic heteroaryl group.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: ring B is a 5-6 membered heterocyclic group containing at least one N; preferably: ring B and ring C are fused to form a fused tricyclic ring, and

is selected from

whereindenotes the chemical bond shared by ring B and ring C; when there is an N atom in the shared chemical bond, the shared chemical bond denoted in ring C is predominant over the shared bond denoted in ring C;

p2 is 0, 1 or 2; and

R_B and p are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein:

ring B and ring C are fused to form a fused tricyclic ring, and

is selected from

whereindenotes the chemical bond shared by ring B and ring C; when there is an N atom in the shared chemical bond, the shared chemical bond denoted in ring C is predominant over the shared bond denoted in ring C.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: ring C is selected from a fused bicyclic ring formed by a 5-membered ring fused to a 6-membered ring and a fused bicyclic ring formed by a 6-membered ring fused to a 6-membered ring; preferably, ring C is selected from a fused bicyclic ring formed by: a 5-membered heterocyclyl fused to a 6-membered aryl, a 5-membered heterocyclyl fused to a 6-membered heteroaryl, a 6-membered heterocyclyl fused to a 6-membered aryl, a 6-membered heterocyclyl fused to a 6-membered heteroaryl, a 5-membered cycloalkyl fused to a 6-membered aryl, a 5-membered cycloalkyl fused to a 6-membered heteroaryl, a 6-membered cycloalkyl fused to a 6-membered aryl, and a 6-membered cycloalkyl fused to a 6-membered heteroaryl; more preferably, ring B and ring C are fused to form a fused tricyclic ring, and

is selected from

whereindenotes the chemical bond shared by ring B and ring C;

q1 is 0, 1, 2 or 3; and

R_C and q are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: ring B and ring C are fused to form a fused tricyclic ring, andis selected from

whereindenotes the chemical bond shared by ring B and ring C.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IC-1-2) ,

wherein:

W₁ is selected from CH₂, NH and O;

W₂ is selected from C, CH and N;

W₃ is selected from CH₂, C=O, NH and O; preferably CH₂, C=O and O;

W₄, W₅, W₆ and W₇ are each independently selected from CH and N; preferably W₄, W₅, W₆, and W₇ are CH;

m₃ is 0 or 1;

m₄ is 0 or 1, preferably 0;

ring A, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above; and

denotes a single bond or a double bond.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: the compound has formula (IC-1-2-1) , formula (IC-1-2-2) , formula (IC-1-2-3) or formula (IC-1-2-4) :
preferably more preferably
preferably more preferably
preferably more preferably
preferably
more preferably

wherein ring A, X₁, X₂, X₃, R_A, R_B, R_C, n, p and q are as defined above;

W₁, W₂, W₃, W₄, W₅, W₆ and W₇ are as defined above;

q1 is 0, 1, 2 or 3; and

p2 is 0, 1 or 2.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: is selected from and

R_A and n are as defined above.
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein: is selected from

n₁ is 0, 1, 2 or 3; and

R_A, X₁, X₂, X₃, R_A, and n are as defined above;

preferably, is selected from
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein the compound has formula (I-1) , formula (IA-1) , formula (IA-1-1) , formula (IA-1-2) , formula (IB-1) , formula (IB-1-1) , formula (IB-1-2) , formula (IC-1) , formula (IC-1-1) or formula (IC-1-2) .
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein the compound is selected from:
The compound according to any preceding claims, or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, wherein the compound is selected from:
A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of Claims 1-30 or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled, and one or more pharmaceutically acceptable carriers or excipients.
Use of a compound according to any one of Claims 1-30 or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled or a pharmaceutical composition according to Claim 31 in the manufacture of a medicament for modulating the hedgehog pathway.
Use of a compound according to any one of Claims 1-30 or a pharmaceutically acceptable salt, geometric isomer, enantiomer, diastereomer, racemate, prodrug, solvate, hydrate thereof or any of the foregoing being isotope-labeled or a pharmaceutical composition according to Claim 31 in the manufacture of a medicament for treating cancer.