WO2025031360A1 - Sulfonamide compound and use thereof - Google Patents

Abstract

A sulfonamide compound represented by formula (I) or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, a preparation method for same and a use of same, and a pharmaceutical composition comprising same. The compound serves as a WDR5 inhibitor, has a remarkable effect as a WDR5 and Myc interaction blocker, and is beneficial to cancer treatment. A¹ and A² each are of a ring structure, and R¹ and R² are substituents.

Description

Sulfonamide compounds and uses thereof Technical Field

The present invention belongs to the field of pharmaceutical chemistry, relates to the technical field of sulfonamide derivatives, and specifically relates to a sulfonamide compound as an anti-neuroblastoma agent, a preparation method and use thereof.

Background Art

It is known that WD40 repeat domain protein 5 (WDR5) belongs to the WD40 protein family and is a protein containing a circular seven-blade β-propeller domain. WDR5 is highly conserved among different vertebrates, with a sequence identity of about 90%. WDR5 was first discovered to be involved in regulating osteoblast differentiation and promoting bone formation during skeletal development. In recent years, more and more studies have revealed the biological functions of WDR5, finding that it not only regulates biological behaviors such as cell proliferation, division, apoptosis, signal transduction, gene transcription, and DNA damage repair under physiological conditions, but is also highly correlated with the occurrence and development of a variety of tumors, making it a popular choice for tumor treatment targets.

The most important function of WDR5 in vivo is epigenetic regulation. As a member of the histone methyltransferase (HMT) complex, it regulates histone methylation levels and activates the transcription of target genes. Abnormal WDR5 function can upregulate the expression of multiple tumor-promoting factors, activate tumor-related signaling pathways, and then enhance tumor cell proliferation, metastasis and drug resistance, driving the occurrence of epithelial-mesenchymal transition (EMT). WDR5 is also an auxiliary factor of myelocytoma viral oncogene homolog (Myc). As a transcription factor, Myc regulates the transcription of downstream genes by binding to the promoter region of the target gene. The limiting factor of this function is the ability of Myc to localize to chromatin. Studies have found that about 80% of the binding of Myc to its downstream target gene sequence requires the participation of WDR5. After the binding of WDR5 and Myc is destroyed, the ability of Myc to induce pluripotent stem cells is greatly weakened. Therefore, WDR5 is a key factor in the process of Myc-driven tumorigenesis. This mechanism has been confirmed in a variety of Myc-related tumors. In glioblastoma and neuroblastoma, WDR5 can promote the binding of Myc to the coactivator protein-associated arginine methyltransferase 1 (CARM1) promoter, enhancing the proliferation and self-renewal ability of tumor cells. In neuroblastoma, WDR5 co-localizes with N-Myc in the double minute homolog gene 2 (MDM2) promoter region, activates the transcription of MDM2, and inhibits the expression of tumor suppressor protein p53. In cholangiocarcinoma, WDR5 maintains a high level of HIF-1α in the body through epigenetic regulation, and also increases the expression of HIF-1α through Myc in the transcription link, promoting the EMT, metastasis and invasion of tumor cells.

There are two interaction sites on the surface of WDR5, namely the WDR5 binding domain (WBM) site and the WDR5 interaction domain (WIN). The WBM site is a shallow and relatively hydrophobic binding pocket, mainly composed of Asn225, Tyr228, Leu240, Phe266, Val268 and Gln289. Retinoblastoma binding protein 5 (RBBP5), C-myc, L-myc, N-myc, KAT8 regulatory NSL complex subunit 2 (KANSL2), etc. all interact with WDR5 through this site. The WIN site is an arginine binding pocket, composed of amino acids such as Ala65, Ser91, Asp107, Phe133, Tyr191, Tyr260 and Phe263. It has been reported that mixed lineage leukemia 1-4 (MLL1-4), KAT8-regulated NSL complex subunit 1 (KANSL1), H3, methylphosphoguanosine phosphate binding protein (MBD3) and kinesin heavy chain member 2A (KIF2A) interact with WDR5 through this site. The key biological functions of WDR5 rely on the mediation of the above two sites, such as direct binding with RBBP5 and SET1/MLL, indirect binding with ash2 (lack, small, or homologous)-like (Drosophila) protein (ASH2L) and dpy-30 homolog protein (DPY30) to assemble into a complex with histone methyltransferase catalytic activity, regulating the transcription of target genes through epigenetic regulation (non-patent document 4); and interacting with the transcription factor Myc, recruiting Myc to chromatin, improving its binding to the promoter region of the target gene, and greatly enhancing its tumor-promoting function (non-patent documents 5,6). Therefore, targeting the WBM or WIN sites on the surface of WDR5 to design specific protein-protein interaction blockers has become the main strategy for the development of new WDR5 inhibitors.

At present, it has been reported that protein-protein interaction blockers designed and developed for the WIN site of WDR5 have strong affinity and good drugability, but their application is limited to the treatment of MLL gene translocation rearrangement leukemia, and they are not effective for solid tumors, while there are few studies on WBM site blockers. Stephen's research team at Vanderbilt University School of Medicine in the United States conducted multiple rounds of optimization and modification, and the obtained compound No. 12 can specifically bind to the WBM site, and the _KD value with WDR5 is 0.10±0.01μM (non-patent literature 7,8), but its anti-tumor effect has not been evaluated.

Therefore, designing and synthesizing highly active compounds that specifically block the interaction between WDR5 and Myc for the treatment of malignant tumors, especially neuroblastoma, has important research significance and application prospects, and is an important direction for potential drug targets and the search for lead compounds.

Non-patent references

1Matthay KK, Maris JM, Schleiermacher G, Nakagawara A, Mackall CL, Diller L, et al. Neuroblastoma. Nature reviews. Disease primers 2016; 2:16078.

2Kholodenko IV, Kalinovsky DV, Doronin, II, Deyev SM, Kholodenko RV. Neuroblastoma Origin and Therapeutic Targets for Immunotherapy. Journal of immunology research 2018; 2018:7394268.

3Migliori V,Mapelli M,Guccione E.On WD40 proteins:propelling our knowledge of transcriptional control? Epigenetics 2012;7:815-22.

4Ernst P, Vakoc CR.WRAD:enabler of the SET1-family of H3K4 methyltransferases.Briefings in functional genomics 2012;11:217-26.

5Thomas LR, Wang Q, Grieb BC, Phan J, Foshage AM, Sun Q, et al. Interaction with WDR5 promotes target gene recognition and tumorigenesis by MYC. Molecular cell 2015;58:440-52.

6Sun Y, Bell JL, Carter D, Gherardi S, Poulos RC, Milazzo G, et al. WDR5 Supports an N-Myc Transcriptional Complex That Drives a Protumorigenic Gene Expression Signature in Neuroblastoma. Cancer research 2015;75:5143-54.

7Chen X, Xu J, Wang X, Long G, You Q, Guo

8Chacón Simon S, Wang F, Thomas LR, Phan J, Zhao B, Olejniczak ET, et al. Discovery of WD Repeat-Containing Protein 5(WDR5)- MYC Inhibitors Using Fragment-Based Methods and Structure-Based Design.Journal of medicinal chemistry 2020;63:4315-33.

Summary of the invention

Based on the above research background, the purpose of the present disclosure is to provide a sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, so as to screen out compounds having excellent performance in terms of effectiveness, safety and selectivity as WDR5 inhibitors.

More specifically, the present disclosure provides a sulfonamide compound represented by formula (I) or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof,

in:

^A1 ring and ^A2 ring are each independently selected from one of a C6-C30 aryl ring, a C6-C30 aliphatic hydrocarbon ring, a C3-C30 heteroaryl ring, and a C3-C30 aliphatic heterocycle, and the "ring" herein includes a monocyclic ring and a polycyclic ring;

R ¹ is a substituent of the A ¹ ring, m=0-3; R ¹ is independently selected from halogen, C3-C5 cycloalkyl, C1-C3 haloalkyl, -CN, C1-C3 alkyl optionally substituted by 1-3 R ^b , C2-C6 alkenyl, C2-C6 alkynyl; R ^b is independently selected from substituted or unsubstituted C1-C3 alkoxy, where "substituted" means optionally substituted by 1-3 hydroxyl, halogen or C ₁ -C ₃ alkoxy substituents;

^R2 is a substituent of the ^A2 ring, n=0-3; ^R2 is selected from -OH, halogen, _C1 - _C3 alkoxy, _C1 - _C3 haloalkyl, -C(O) ^NHRc , -NHC(O) _CH2NHC (O) ^Rc , 3-5 membered cycloalkyl optionally substituted by 1-3 "-CN", 5-7 membered aryl optionally substituted by 1-3 "-CN"; ^Rc is _C1 - _C3 alkyl;

L is a chemical bond or a divalent group selected from one or more combinations of C1-6 alkylene, saturated or partially unsaturated C3-10 cycloalkylene, -O-, -NR ^a -, and -NR ^a -C1-6 alkylene; ^Ra is independently selected from H, C1-C10 alkyl, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated 3-10 membered heterocyclic group or C6-10 aryl, CH ₂ in ^Ra can be replaced by -O- or -S-, and H in ^Ra can be replaced by hydroxyl, halogen or C1-C3 alkoxy.

The compounds disclosed herein are compounds that specifically bind to the WBM site of WDR5, which can block the interaction between WDR5 and Myc, thereby regulating Myc-mediated oncogene transcription and achieving tumor treatment effects. In pharmacological experiments, it was shown that the compounds disclosed in the present invention have a strong affinity for WDR5 protein. In neuroblastoma cell lines with high MYCN amplification, the compounds can significantly inhibit cell proliferation, while having no significant effect on the survival of human embryonic kidney cells HEK293T, showing good selectivity and safety. According to the results of crystal complex structure analysis, the representative compounds disclosed herein can specifically bind to the WBM domain of WDR5, blocking the interaction between WDR5 and Myc, thereby exerting anti-tumor activity.

In an optional embodiment of the present disclosure, ^A1 ring represents a cyclic group selected from 5-7 membered cycloalkyl, 5-7 membered cycloalkenyl, 5-7 membered aryl, 5-7 membered heterocyclyl, 5-7 membered heteroaryl, ^A2 ring represents a cyclic group selected from 5-7 membered cycloalkyl, 5-7 membered cycloalkenyl, 5-7 membered aryl, 5-7 membered heterocyclyl, 5-10 membered heteroaryl; at least one halogen is present in ^R1 and ^R2 as a substituent.

In an optional embodiment of the present disclosure, the ^A1 ring or the ^A2 ring is one of the following cyclic structures:

* indicates the site of connection with the mother core, and ^R1 and ^R2 as substituents can replace any site that is chemically substitutable.

In an optional embodiment of the present disclosure, ^R1 or ^R2 is selected from H, F, Cl, Br, I, hydroxyl, methyl, methoxy, trifluoromethyl, ethyl, ethynyl, cyclopropyl, cyclopentyl, cyclohexyl, cyano, formamide, cyano-substituted cyclopropyl, cyano-substituted cyclopentyl, cyano-substituted cyclohexyl, phenyl, cyano-substituted phenyl, naphthyl, cyano-substituted naphthyl and one of the following groups:

* indicates the site of attachment to the parent nucleus.

In an optional embodiment of the present disclosure, the combination of the ^A1 ring and its substituent ^R1 is one selected from the following groups:

In an optional embodiment of the present disclosure, the combination of the ^A2 ring and its substituent ^R2 is one selected from the following groups:

* indicates the site of attachment to the parent nucleus.

The compounds disclosed herein can be used as WDR5-Myc interaction blockers. Specifically, the pharmacological experimental results show that the compounds disclosed herein can specifically bind to the WBM domain of WDR5 and block the interaction between WDR5 and Myc. Based on this use, the compounds disclosed herein can be used in the preparation of drugs for treating diseases related to abnormal activation of Myc, RBBP5, KANSL2 and other related signaling pathways caused by abnormal expression of WDR5.

It is well known that diseases related to abnormal activation of Myc, RBBP5, KANSL2 and other related signaling pathways caused by abnormal expression of WDR5 include but are not limited to neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, ganglioneuroma, sympathetic neuroblastoma, neurilemmoma, neurofibroma, prostate cancer, triple-negative breast cancer, nasopharyngeal cancer, esophageal cancer, laryngeal cancer, lung cancer, gastric cancer, liver cancer, colorectal cancer, cervical cancer, pancreatic cancer, bladder cancer, retinoblastoma, osteosarcoma, chondrosarcoma, chordoma, rhabdomyosarcoma, multiple myeloma, lymphoma, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia and other diseases, especially suitable for neurogenic tumors including: neuroblastoma, gangliocytoma, ganglioblastoma, Treatment and relief drugs for ganglioneuroma, ganglioneuroma, sympathetic neuroblastoma, schwannoma, and neurofibroma.

Representative compounds of the present invention include the following compounds, but are not limited thereto:

Another aspect of the present disclosure provides a pharmaceutical composition, characterized in that the pharmaceutical composition comprises the sulfonamide compound disclosed above or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, and a pharmaceutically acceptable carrier and/or excipient, and the pharmaceutical composition can be a solid preparation, a semisolid preparation, a liquid preparation or a gaseous preparation;

The dosage form of the pharmaceutical composition can be an oral dosage form or an injection, and the oral dosage form includes capsules, tablets, pills, powders and granules. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures, and the injection contains physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions.

In the present disclosure, a "pharmaceutically acceptable" ingredient is a substance that is suitable for use in humans and/or animals without excessive adverse side effects (such as toxicity, irritation, and allergic response) at a reasonable benefit/risk ratio.

In the present disclosure, "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or excipient used to deliver the active substance of the present disclosure or a physiologically acceptable salt thereof to animals or humans. The carrier may be liquid or solid.

In the present disclosure, the pharmaceutical composition contains a safe and effective amount (such as 0.001-99.9 parts by weight, 0.01-99 parts by weight, or 0.1-90 parts by weight) of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 parts by weight.

Alternatively, the pharmaceutical composition disclosed herein contains 0.001-99.9 wt %, more preferably 0.01-99 wt %, and more preferably 0.1-90 wt % of the total weight of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 wt %.

In another optional embodiment, the optional ratio of the compound of formula (I) to the pharmaceutically acceptable carrier, excipient or sustained-release agent is that the compound of formula (I) as the active ingredient accounts for more than 70% of the total weight, and the remaining part accounts for 0.5-30% of the total weight, or better 1-15%, or most preferably 2-10%.

The various formulation forms of the pharmaceutical composition disclosed herein have a unit dose containing 0.5 mg-200 mg, 2 mg-100 mg, or even 5 mg-50 mg of the compound of formula (I), enantiomers, racemates, pharmaceutically acceptable salts or mixtures thereof.

When the pharmaceutical composition contains an additional active pharmaceutical ingredient for treating or preventing cancer, the dosage of the active ingredient can generally be the conventional dosage in the prior art or lower.

The pharmaceutical composition of the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, etc., wherein the compound of formula (I) can be present in a suitable solid or liquid carrier or diluent. The pharmaceutical composition of the present invention can also be stored in a suitable sterilizing apparatus for injection or instillation. The pharmaceutical composition can also contain odorants, flavoring agents, etc.

The compound of formula (I) disclosed herein or the pharmaceutical composition comprising the compound of formula (I) can be used clinically in mammals (including humans) through administration routes such as oral, nasal, skin, lung or gastrointestinal tract. The optional administration route is oral. The optional daily dose is 0.5mg-100mg/kg body weight, taken once or in divided doses. Regardless of the method of administration, the optimal dose for an individual should be determined according to the specific treatment. Usually, it is started with a small dose and the dose is gradually increased until the most suitable dose is found.

The effective dose of the active ingredient used may vary with the compound used, the mode of administration, and the severity of the disease to be treated. However, generally, satisfactory results can be obtained when the compounds of the present disclosure are administered at a dose of about 1-100 mg/kg of animal body weight per day, preferably in 1-3 divided doses per day, or in a sustained release form. For most large mammals, the total daily dose is about 5-500 mg, preferably about 10-250 mg. A dosage form suitable for oral administration comprises about 1-100 mg of active compound closely mixed with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen can be adjusted to provide the best therapeutic response. For example, several divided doses may be administered per day, or the dose may be reduced proportionally, depending on the urgency of the treatment condition.

The compound or its pharmaceutically acceptable salt and composition thereof can be administered orally as well as intravenously, intramuscularly or subcutaneously. From the standpoint of ease of preparation and administration, the optional pharmaceutical composition is a solid composition, especially a tablet and a solid-filled or liquid-filled capsule. Oral administration of the pharmaceutical composition is optional.

Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include sterile water, polyethylene glycol, nonionic surfactants and edible oils (such as corn oil, peanut oil and sesame oil), as long as they are suitable for the characteristics of the active ingredient and the specific administration method required. Adjuvants commonly used in the preparation of pharmaceutical compositions may also be advantageously included, such as flavoring agents, pigments, preservatives and antioxidants such as vitamin E, vitamin C, BHT and BHA.

The active compounds or their pharmaceutically acceptable salts and compositions thereof may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds (as free bases or pharmaceutically acceptable salts) may also be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquids, polyethylene glycols and mixtures thereof in oils. Under conventional storage and use conditions, these preparations contain preservatives to prevent the growth of microorganisms.

The pharmaceutical forms adapted for injection include: sterile aqueous solutions or dispersions and sterile powders (for the temporary preparation of sterile injection solutions or dispersions). In all cases, these forms must be sterile and must be fluid to facilitate the discharge of fluid from a syringe. Must be stable under manufacturing and storage conditions, and must be able to prevent the contamination effects of microorganisms (such as bacteria and fungi). The carrier can be a solvent or dispersion medium, which contains, for example, water, alcohol (such as glycerol, propylene glycol and liquid polyethylene glycol), their appropriate mixtures and vegetable oils.

The compound represented by formula (I) or its pharmaceutically acceptable salt and the composition thereof can also be administered in combination with other active ingredients or drugs for treating or preventing cancer diseases. When two or more drugs are administered in combination, it generally has a better effect than the two drugs administered separately.

Another aspect of the present disclosure provides the use of the compounds of the present disclosure and the corresponding pharmaceutical compositions in the preparation of cancer treatment or relief drugs. The compositions and methods provided by the present disclosure can be used to treat a variety of cancers, including prostate, breast, brain, skin, cervical cancer, testicular cancer, etc. More specifically, the cancers that can be treated by the compositions and methods of the present disclosure include, but are not limited to, tumor types, such as astrocytic carcinoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, stomach, head and neck, hepatocellular carcinoma, laryngeal cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer and thyroid cancer and sarcoma. More specifically, these compounds are useful in the treatment of: Heart: sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma; Lung: bronchial carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagon, gastrinoma, carcinoid, VIPoma), small intestine (adenocarcinoma, lymphoma, carcinoid, Kaposi's sarcoma, leiomyosarcoma, hemangioma, lipoma), Urogenital tract: Kidney (adenocarcinoma, Wilms' tumor (Nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); Liver: Hepatocellular carcinoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma ; Biliary tract: Gallbladder cancer, ampullary cancer, bile duct cancer; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, chronic bone tumor (osteochondrosarcoma), benign tumors, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumor; Nervous system: skull (osteoma, hemangioma, granuloma, xanthomas, osteitis), meninges (meningioma, meningosarcoma, glioma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, reproductive Gynecological: uterus (endometrial cancer), cervix (cervical cancer, precancerous cervical dysplasia), ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell tumor, supporting cell tumor, germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, cystoid sarcoma) (embryonal rhabdomyosarcoma), fallopian tube (cancer); hematology: blood (acute and chronic myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus, lipoma, hemangioma, dermatofibroma, lupus erythematosus, psoriasis and adrenal gland: neuroblastoma. In certain embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL).

In an optional embodiment, a compound having a structure represented by the following formula (I) or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof is provided for use as a drug for treating neurogenic tumors. The present disclosure also provides a compound having a structure represented by the following formula (I) or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof as a drug for treating and alleviating neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, ganglioblastoma, sympathetic neuroblastoma, schwannoma, and neurofibroma.

In an optional embodiment, the cancer is selected from neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, sympathetic neuroblastoma, neurotheca, neurofibroma, prostate cancer, triple-negative breast cancer, nasopharyngeal carcinoma, esophageal cancer, laryngeal cancer, lung cancer, gastric cancer, liver cancer, colorectal cancer, cervical cancer, pancreatic cancer, bladder cancer, retinoblastoma, osteosarcoma, chondrosarcoma, chordoma, rhabdomyosarcoma, multiple myeloma, lymphoma, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia; in an optional embodiment, the cancer is selected from neurogenic tumors including: neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, sympathetic neuroblastoma, neurotheca, neurofibroma.

The present disclosure also provides a method for treating and/or preventing cancer, which comprises administering a therapeutically effective amount of the sulfonamide compound of the present disclosure or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, or the pharmaceutical composition thereof to a human or animal body.

Explanation of terms

Unless otherwise stated, some of the terms used in the specification and claims of this disclosure are defined as follows:

definition

Unless otherwise defined below, the meanings of all technical and scientific terms used herein are intended to be the same as those generally understood by those skilled in the art. Reference to the technology used herein is intended to refer to the technology generally understood in the art, including those changes in technology or replacement of equivalent technology that are obvious to those skilled in the art. Although it is believed that the following terms are well understood by those skilled in the art, the following definitions are still set forth to better explain the present disclosure.

In the present disclosure, the terms "includes," "comprising," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude other unrecited elements or method steps.

As used herein, the term "alkylene" refers to a saturated divalent hydrocarbon group, and may refer to a saturated divalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene, ethylene, propylene or butylene.

The aliphatic cyclic group refers to a cycloalkyl, cycloalkenyl, cycloalkynyl, etc. composed of a ring structure of C and H, wherein C is replaced by a heteroatom to form an aliphatic heterocyclic group; it also includes the bridged ring and spiro ring forms described later.

As used herein, the term "alkyl" is defined as a linear or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl has 1 to 12, for example 1 to 6 carbon atoms. For example, as used herein, the term "C1-6 alkyl" refers to a linear or branched group of 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or n-hexyl), which is optionally substituted by 1 or more (such as 1 to ₃ ) suitable substituents such as halogen (in this case, the group is referred to as "haloalkyl") (e.g., _CH2F , _CHF2 , _CF3 , _CCl3 , _{C2F5 , C2Cl5} , _{CH2CF3 , CH2Cl} or _{-CH2CH2CF3 , etc.} ). The term "C1-4 alkyl" refers to a linear or branched aliphatic hydrocarbon chain of 1 to 4 carbon atoms (ie, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl).

As used herein, the term "alkenyl" means a linear or branched monovalent hydrocarbon radical containing one double bond and having 2 to 6 carbon atoms ("C The alkenyl group is, for example, vinyl, ₁ -propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl and 4-methyl-3-pentenyl. When the compounds of the present disclosure contain an alkenyl group, the compounds may be present in the pure E (entgegen) form, the pure Z (zusammen) form or any mixture thereof.

As used herein, the term "alkynyl" refers to a monovalent hydrocarbon group containing one or more triple bonds, which may be of 2, 3, 4, 5 or 6 carbon atoms, such as ethynyl or propynyl.

As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., a monocyclic ring such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or a bicyclic ring including spiro, fused or bridged systems (such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, decalinyl, etc.), optionally substituted with 1 or more (such as 1 to 3) suitable substituents. The cycloalkyl has 3 to 15 carbon atoms. For example, the term " _C3-6 cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring of 3 to 6 ring carbon atoms (such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), which is optionally substituted by 1 or more (such as 1 to 3) suitable substituents, such as methyl substituted cyclopropyl.

As used herein, the terms "cycloalkylene", "cycloalkyl" and "hydrocarbon ring" refer to a saturated (i.e., "cycloalkylene" and "cycloalkyl") or unsaturated (i.e., having one or more double bonds and/or triple bonds within the ring) monocyclic or polycyclic hydrocarbon ring having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring carbon atoms, including but not limited to (cyclo)propyl (ring), (cyclo)butyl (ring), (cyclo)pentyl (ring), (cyclo)hexyl (ring), (cyclo)heptyl (ring), (cyclo)octyl (ring), (cyclo)nonyl (ring), (cyclo)hexenyl (ring) and the like.

As used herein, the terms "heterocyclyl", "heterocyclylene" and "heterocycle" refer to a saturated (i.e., heterocycloalkyl) or partially unsaturated (i.e., having one or more double bonds and/or triple bonds within the ring) cyclic group having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring atoms, wherein at least one ring atom is a heteroatom selected from N, O and S and the remaining ring atoms are C. For example, a "3-10 membered (sub)heterocyclyl" is a saturated or partially unsaturated (sub)heterocyclyl having 2-9 (e.g., 2, 3, 4, 5, 6, 7, 8 or 9) ring carbon atoms and one or more (e.g., 1, 2, 3 or 4) heteroatoms independently selected from N, O and S. Examples of heterocyclylene and heterocyclyl include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, dioxolinyl, pyrrolidinyl, pyrrolidonyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl. The group also encompasses bicyclic systems, including spiro, fused or bridged systems (such as 8-azaspiro[4.5]decane, 3,9-diazaspiro[5.5]undecane, 2-azabicyclo[2.2.2]octane, etc.). The heterocyclylene and heterocyclyl groups may be optionally substituted with one or more (e.g., 1, 2, 3 or 4) suitable substituents.

As used herein, the terms "(ylidene)aryl" and "aromatic ring" refer to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π electron system. For example, as used herein, the terms "C _6-10 (ylidene)aryl" and "C _6-10 aromatic ring" mean an aromatic group containing 6 to 10 carbon atoms, such as (ylidene)phenyl (benzene ring) or (ylidene)naphthyl (naphthalene ring). The (ylidene)aryl group and the aromatic ring are optionally substituted with 1 or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO ₂ , C _1-6 alkyl, etc.).

As used herein, the terms "heteroaryl(ene)" and "heteroaromatic ring" refer to a monocyclic, bicyclic or tricyclic aromatic ring system having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and which contains at least one heteroatom which may be identical or different (the heteroatom being, for example, oxygen, nitrogen or sulfur) and, in each case additionally may be benzo-fused. In particular, "heteroaryl" or "heteroaromatic ring" is selected from thiophenyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, etc., and benzo derivatives thereof; or pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and benzo derivatives thereof.

As used herein, the term "aralkyl" preferably refers to an alkyl substituted with an aryl or heteroaryl, wherein the aryl, heteroaryl and alkyl are as defined herein. Typically, the aryl may have 6-14 carbon atoms, the heteroaryl may have 5-14 ring atoms, and the alkyl may have 1-6 carbon atoms. Exemplary aralkyls include, but are not limited to, benzyl, phenylethyl, phenylpropyl, phenylbutyl.

As a more specific explanation of the terms are as follows:

"Alkyl" refers to a saturated aliphatic hydrocarbon group, comprising 1-20 carbon atoms, or 1-10 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms, or 1-2 carbon atoms, a saturated straight or branched monovalent hydrocarbon group, wherein the alkyl group may be independently optionally substituted with one or more substituents described in the present disclosure. Further examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group may be optionally substituted or unsubstituted.

"Alkenyl" refers to a linear or branched monovalent hydrocarbon group of 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one CC is an ^sp2 double bond, wherein the alkenyl group may be independently optionally substituted with one or more substituents described in the present disclosure, wherein specific examples include, but are not limited to, vinyl, allyl, and butylene, etc. Alkenyl may be optionally substituted or unsubstituted.

"Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, the cycloalkyl ring comprising 3 to 20 carbon atoms, preferably comprising 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyl includes cycloalkyl of spiro ring, condensed ring and bridge ring. Cycloalkyl can be optionally substituted or unsubstituted.

"Spirocycloalkyl" refers to a polycyclic group with 5 to 18 members, two or more cyclic structures, and one carbon atom (called spiro atom) shared between the monocyclic rings, containing one or more double bonds in the ring, but no ring has a completely conjugated π electron aromatic system. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. According to the number of spiro atoms shared between the rings, the spirocycloalkyl is divided into single spiro, double spiro or multi-spirocycloalkyl, preferably single spiro and double spirocycloalkyl, preferably 4/5 members, 4/6 members, 5/5 members or 5/6 members. Non-limiting examples of "spirocycloalkyl" include, but are not limited to:

"Fused cycloalkyl" refers to a 5 to 18-membered, all-carbon polycyclic group containing two or more cyclic structures sharing a pair of carbon atoms, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic alkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to:

"Bridged cycloalkyl" refers to a 5 to 18-membered, all-carbon polycyclic group containing two or more cyclic structures, sharing two carbon atoms that are not directly connected to each other, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to:

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl and the like.

"Heterocyclyl", "heterocycle" or "heterocyclic" are used interchangeably in this application and refer to a saturated or partially unsaturated monocyclic, bicyclic or tricyclic non-aromatic heterocyclic group containing 3-12 ring atoms, wherein at least one ring atom is a heteroatom, such as an oxygen, nitrogen, sulfur atom, etc. Preferably, it has a 5-7 membered monocyclic ring or a 7-10 membered bi- or tricyclic ring, which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and piperazinyl. The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is the heterocyclyl. The heterocyclyl may be optionally substituted or unsubstituted.

"Spiro heterocyclyl" refers to a polycyclic group with 5 to 18 members, two or more ring structures, and one atom shared between the monocyclic rings, containing one or more double bonds in the ring, but no ring has a completely conjugated π-electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen, sulfur or S(O) _m heteroatoms, and the remaining ring atoms are carbon, m = 1 or 2. Preferably 6 to 14 members, more preferably 7 to 10 members. According to the number of spiro atoms shared between rings, spiro heterocyclyl is divided into single spiro heterocyclyl, double spiro heterocyclyl or multi-spiro heterocyclyl, preferably single spiro heterocyclyl and double spiro heterocyclyl. More preferably 4/4, 4/5, 4/6, 5/5 or 5/6 monospiro heterocyclyl. Non-limiting examples of "spiro heterocyclyl" include, but are not limited to:

"Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more cyclic structures that share a pair of atoms, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π-electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen, sulfur or S(O) _m heteroatoms, and the remaining ring atoms are carbon, m = 1 or 2. Preferably 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of "fused heterocyclic group" include, but are not limited to:

"Bridged heterocyclic group" refers to a polycyclic group with 5 to 18 members, containing two or more cyclic structures, sharing two atoms that are not directly connected to each other, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π-electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen, sulfur or S(O) _m heteroatoms, and the remaining ring atoms are carbon, m = 1 or 2. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclic groups" include, but are not limited to:

"Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be connected together in a fused manner. The term "aryl" includes aromatic groups such as phenyl, naphthyl, and tetrahydronaphthyl. Preferably, the aryl is a C ₆ -C ₁₀ aryl, more preferably, the aryl is phenyl and naphthyl, and most preferably, phenyl. The aryl may be substituted or unsubstituted. The "aryl" may be fused with a heteroaryl, a heterocyclic group, or a cycloalkyl group, wherein the aryl ring is connected to the parent structure, and non-limiting examples include, but are not limited to:

"Heteroaryl" refers to an aromatic 5 to 6-membered monocyclic or 9 to 10-membered bicyclic ring, which may contain 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. The embodiment of "heteroaryl" includes, but is not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzodioxolyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolyl, indazolyl, benzisothiazolyl, benzoxazolyl and benzisoxazolyl. Heteroaryl may be optionally substituted or unsubstituted. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples include but are not limited to:

"Alkoxy" refers to a group of (alkyl-O-), wherein alkyl is as defined herein. _{C 1} -C ₆ alkoxy is preferred, and examples thereof include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

"Haloalkyl" refers to an alkyl group having one or more halogen substituents, wherein the alkyl group has the meaning as described in the present disclosure. Examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, perfluoroethyl, 1,1-dichloroethyl, 1,2-dichloropropyl, and the like.

"Carboxylate" refers to -C(O)O(alkyl) or (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms and at least one nitrogen atom in the ring, which may optionally contain one or more (e.g., one, two, three or four) ring members selected from N, O, C=O, S, S=O and S(=O) ₂ , which is connected to the rest of the molecule via the nitrogen atom in the nitrogen-containing heterocycle and any remaining ring atoms, and the nitrogen-containing heterocycle is optionally benzo-fused and can be connected to the rest of the molecule via the nitrogen atom in the nitrogen-containing heterocycle and any carbon atom in the fused benzene ring.

The term "substituted" means that one or more (e.g., one, two, three, or four) hydrogens on the designated atom are replaced by a selection from the indicated group, provided that the normal valence of the designated atom in the present context is not exceeded and the substitution forms a stable compound. Combinations of substituents and/or variables are permitted only if such combinations form stable compounds.

If a substituent is described as "optionally substituted," the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as optionally substituted with one or more of the substituents listed, one or more hydrogens on the carbon (to the extent any hydrogens are present) may be substituted individually and/or in combination. If a nitrogen of a substituent is described as being optionally substituted with one or more of the substituent lists, then one or more hydrogens on the nitrogen (to the extent any hydrogens are present) may each be replaced with an independently selected optional substituent.

If substituents are described as being "independently selected" from a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

The present disclosure also includes all pharmaceutically acceptable isotopically labeled compounds, which are identical to the compounds of the present disclosure except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of isotopes suitable for inclusion in the compounds of the present disclosure include, but are not limited to, isotopes of hydrogen (e.g., deuterium ( ^2H ), tritium ( ^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 disclosure (e.g., those incorporating radioisotopes) are useful in drug and/or substrate tissue distribution studies (e.g., assays). The radioisotopes tritium (i.e., ³ H) and carbon-14 (i.e., 14 C) are particularly useful for this purpose because they are easily incorporated and easily detected. Substitution with positron emitting isotopes (e.g., ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N) can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the present disclosure can be prepared by methods similar to those described in the accompanying routes and/or examples and preparations by using appropriate isotopically labeled reagents in place of the non-labeled reagents previously employed. Pharmaceutically acceptable solvates of the present disclosure include those in which the crystallization solvent may be isotopically substituted, for example, D ₂ O, acetone-d ₆ , or DMSO-d ₆ .

"Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1-3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.

As used herein, “substituted” or “substituted”, unless otherwise specified, means that a group may be substituted by one or more groups selected from the following: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =O, -C(O) ^Rb , ^-OC (O) ^Rb , ^-NRbRb , ^-C (O) ^NRbRb , -NRbC(O) ^{Rb , -S} (O) ^NRbRb or -S(O ⁾ _2NRbRb ^, wherein ^Rb is as defined in the general formula (I).

As used herein, a "therapeutically effective dose" of a compound is an amount sufficient to ameliorate or in some way reduce symptoms, halt or reverse the progression of a condition. Such a dose may be used as a single dose or may be taken according to a regimen to be effective. As used herein, "treat" means to improve or otherwise alter the symptoms or pathology of a patient's condition, disorder or disease in any way.

As used herein, "amelioration of the symptoms of a particular disease by administration of a particular compound or pharmaceutical composition" refers to any reduction, whether permanent or temporary, lasting or transitory, attributable to or associated with the administration of that composition.

The compounds of the present disclosure may contain asymmetric centers or chiral centers and therefore exist in different stereoisomers. All stereoisomeric forms of the compounds of the present disclosure, including but not limited to, diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, constitute part of the present disclosure. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, lacking optical activity.

"Tautomers" or "tautomeric forms" refer to structural isomers of different energies that are interconvertible via a low energy barrier. Single stereochemical isomers of the disclosed compounds or mixtures of their enantiomers, diastereomers, or geometric isomers are within the scope of the present disclosure.

In this article, solid lines can be used Solid wedge Virtual wedge Depict the chemical bonds of the compounds of the present disclosure. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the stereoisomer shown is present. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present disclosure are intended to exist in the form of stereoisomers, which include cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, atropisomers, and mixtures thereof. The compounds of the present disclosure may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereoisomer pairs).

The present disclosure encompasses all possible crystalline forms or polymorphs of the compounds of the present disclosure, which may be a single polymorph or a mixture of more than one polymorph in any ratio.

It should also be understood that certain compounds of the present disclosure may be used in free form for treatment, or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present disclosure, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites, chelates, complexes, inclusion compounds or prodrugs, which, after being administered to a patient in need thereof, are capable of directly or indirectly providing Therefore, when referring to "compounds of the present disclosure" herein, the above-mentioned various derivative forms of the compounds are also intended to be covered.

Pharmaceutically acceptable salts of the compounds of the present disclosure include acid addition salts and base addition salts thereof, including but not limited to salts containing hydrogen bonds or coordinate bonds.

Suitable acid addition salts are formed from acids which form pharmaceutically acceptable salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate/carbonate, bisulfate/sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hyaluronate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogenphosphate/dihydrogenphosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, toluenesulfonate, trifluoroacetate, and xinofoate.

Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, benzathine penicillin salts, calcium salts, choline salts, diethylamine salts, diethanolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, ethanolamine salts, potassium salts, sodium salts, tromethamine salts, and zinc salts. Methods for preparing pharmaceutically acceptable salts of compounds of the present disclosure are known to those skilled in the art.

As used herein, the term "ester" means an ester derived from the compounds of the general formula in the present application, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present disclosure in free acid or alcohol form). The compounds of the present disclosure themselves can also be esters.

The compounds of the present disclosure may exist in the form of solvates (preferably hydrates), wherein the compounds of the present disclosure contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of the polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides, as nitrogen requires an available lone pair of electrons to oxidize to an oxide; those skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art, including oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and meta-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxirane such as dimethyldioxirane.

Also included within the scope of the present disclosure are metabolites of the compounds of the present disclosure, i.e., substances formed in vivo upon administration of the compounds of the present disclosure. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Thus, the present disclosure includes metabolites of the compounds of the present disclosure, including compounds produced by a process of contacting the compounds of the present disclosure with a mammal for a period of time sufficient to produce a metabolic product thereof.

The present disclosure further includes within its scope prodrugs of compounds of the present disclosure, which are certain derivatives of compounds of the present disclosure that may themselves have little or no pharmacological activity, but which when administered into or on the body may be converted, for example, by hydrolytic cleavage, into compounds of the present disclosure having the desired activity. Typically such prodrugs will be functional group derivatives of the compounds that are easily converted into the desired therapeutically active compound in vivo.

The present disclosure also encompasses compounds of the present disclosure containing protecting groups. In any process for preparing the compounds of the present disclosure, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present disclosure. This can be achieved by conventional protecting groups, for example, those described in T.W.Greene & P.G.M.Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups may be removed at an appropriate subsequent stage using methods known in the art.

The term "about" means within ±10% of the stated numerical value, preferably within ±5%, more preferably within ±2%.

The present disclosure provides a WDR5/Myc interaction blocker with a new structure. Experimental results show that this type of derivative exhibits excellent anti-neuroblastoma activity, as well as excellent safety and selectivity, and can be used to prepare drugs for treating cancer, especially neuroblastoma and other diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a schematic diagram of the crystal structure of the complex of the disclosed compound and WDR5.

DETAILED DESCRIPTION

The method of the present disclosure is described below by specific examples to make the technical solution of the present disclosure easier to understand and grasp, but the present disclosure is not limited thereto. In the following examples, ¹ H NMR spectra were measured using a Bruker instrument (400 MHz), and chemical shifts were expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. ¹ H NMR expression method: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad, dd = doublet of doublets, dt = doublet of triplet. If coupling constants are provided, the unit is Hz.

Mass spectra were obtained using LC/MS instrument, and the ionization mode was ESI. High performance liquid chromatograph model: Agilent 1260, Thermo Fisher U3000; chromatographic column model: Waters xbrige C18 (4.6*150mm, 3.5μm); mobile phase: A: ACN, B: Water (0.1% H ₃ PO ₄ ); flow rate: 1.0mL/min; gradient: 5% A for 1min, increase to 20% A within 4min, increase to 80% A within 8min, 80% A for 2min, back to 5% A within 0.1min; wavelength: 220nm; column oven: 35℃. The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) uses a specification of 0.2mm-0.3mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm-0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available materials and reagents are used directly without further purification. Unless otherwise specified, commercial manufacturers include but are not limited to Sinopharm Group, J&K Technology Co., Ltd., TCI (Shanghai) Chemical Industry Development Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd. and Shanghai Myrel Chemical Technology Co., Ltd. CD ₃ OD: deuterated methanol; CDCl ₃ : deuterated chloroform; DMSO-d ₆ : deuterated dimethyl sulfoxide; Pd ₂ (dba) ₃ : tris(dibenzylideneacetone)dipalladium; Pd(dppf)Cl ₂ ：[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium; XantPhos: 4,5-bis(diphenylphosphino)ferrocene-9,9-dimethylxanthene; XPhos: 2-dicyclohexylphospho-2,4,6-triisopropylbiphenyl; HATU: 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate; EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; HOBT: 1-hydroxybenzotriazole; BINAP: 1,1'-binaphthyl-2,2'-bis(diphenylphosphine); DCM: dichloromethane; PE: petroleum ether; EA: ethyl acetate; MeOH: methanol; DMF: N,N-dimethylformamide; TLC: thin layer chromatography; HPLC: high performance liquid chromatography; purity: purity; R _f : The ratio of the distance from the origin to the center of the spot to the distance from the origin to the solvent front in thin layer chromatography. ; Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a volume of about 1L.

Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution. Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20°C-30°C. The monitoring of the reaction progress in the examples is performed by thin layer chromatography (TLC), and the developing solvent used in the reaction, the system of eluents for column chromatography or the developing solvent system for thin layer chromatography used for purifying compounds include: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane: ethyl acetate; wherein the volume ratio of the solvent varies according to the polarity of the compound, and a small amount of acidic or alkaline reagents may also be added for adjustment, such as acetic acid or triethylamine. The reagent for providing alkaline conditions is selected from organic bases or inorganic bases, the organic base is one or more of triethylamine, N,N-diisopropylethylamine, n-butyl lithium, diisopropyl lithium amide, bistrimethylsilyl lithium amide, sodium tert-butoxide, sodium methoxide and potassium tert-butoxide, the inorganic base is one or more of sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium bicarbonate and lithium hydroxide; the reagent for providing acidic conditions is one or more of hydrogen chloride, 1,4-dioxane solution of hydrogen chloride, methanol solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid and phosphoric acid;

The metal catalyst is one or more of palladium/carbon, Raney nickel, tetrakistriphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (Pd(dppf)Cl ₂ ), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, bistriphenylphosphine palladium dichloride (Pd(PPh ₃ ) ₂ Cl ₂ ) and tris(dibenzylideneacetone)dipalladium (Pd ₂ (dba) ₃ );

The ligand is one or more of 2-dicyclohexylphosphino-2,6'-dimethoxybiphenyl (SPhos), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (XantPhos), 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (XPhos), 2-dicyclohexylphosphino-2'-(N,N-dimethylamino)-biphenyl (DavePhos), 1,1'-bis(diphenylphosphino)ferrocene (Dppf) and 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (BINAP), preferably 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (BINAP);

The reducing agent is one or more of sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, and lithium aluminum tetrahydride;

The oxidant is one or more of potassium permanganate, manganese dioxide, potassium dichromate, sodium dichromate and potassium osmate;

The above reaction is preferably carried out in a solvent, and the solvent used is one or more of N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, 1,4-dioxane, water, tetrahydrofuran, dichloromethane, 1,2-dichloroethane, methanol, ethanol, toluene, petroleum ether, ethyl acetate, n-hexane and acetone.

Pharmaceutical Chemical Synthesis

Preparation of intermediates

Intermediate 1

2,4-Dichlorobenzoyl chloride IN-1

Step 1 2,4-Dichlorobenzoyl chloride IN-1

N,N-dimethylformamide (3 drops) was slowly added dropwise to a mixture of 2,4-dichlorobenzoic acid IN-1a (300 mg, 1.57 mmol) and oxalyl chloride (3 mL, 35.5 mmol) in anhydrous tetrahydrofuran (15 mL) at 0°C, and reacted at room temperature for 2 hours. TLC showed that the starting material disappeared. The reaction solution was directly concentrated to obtain the title compound IN-1 (320 mg, crude product) as a white solid, which was directly used in the next step.

Example 1

2,4-Dichloro-N-((2,4-dimethylthiazol-5-yl)sulfonyl)benzamide 1

Step 1 2,4-Dimethyl-1,3-thiazole-5-sulfonyl chloride 1b

2,4-Dimethylthiazole 1a (5.00 g, 44.2 mmol) was slowly added dropwise to chlorosulfonic acid (10 mL) at room temperature, and the temperature was raised to 140 ° C for 18 hours. The reaction solution naturally cooled to room temperature, and phosphorus pentachloride (18.4 g, 88.4 mmol) was slowly added. There was a violent exotherm (pay attention to prevent spraying). After the addition, the temperature was raised to 120 ° C, and the reaction was continued for 3 hours. TLC showed that the raw material disappeared. The reaction solution cooled to room temperature and was slowly added dropwise to ice water (100 mL). Dichloromethane (20 mL x 3) was added for extraction, and the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 1b (9.3 g, crude product) as a brown oil, which was directly used in the next step.

Step 2 2,4-Dimethylthiazole-5-sulfonamide 1c

Compound 1b (8.8 g, crude product) was dissolved in 1,4-dioxane (50 mL), cooled to 0°C, and ammonia water (15 mL) was slowly added dropwise. After the addition was completed, the reaction was continued at 0°C for 10 minutes. TLC showed that the raw material disappeared. The reaction solution was slowly poured into ice water (100 mL), and diluted hydrochloric acid (3N) was added to acidify to pH = 3. Dichloromethane (30 mL x 3) was added for extraction. The organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound 1c (7.2 g, crude product) as a brown oily liquid, which was directly used in the next step.

Step 3 2,4-dichloro-N-((2,4-dimethylthiazol-5-yl)sulfonyl)benzamide 1

Compound 1c (230 mg, crude product) was dissolved in ethyl acetate (10 mL), and 4-dimethylaminopyridine (5 mg, 0.04 mmol), triethylamine (2 mL, 14.4 mmol) and toluene solution (15 mL) of intermediate IN-1 (320 mg, crude product) were added in sequence at room temperature. After addition, the mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 1 (116 mg, three-step yield 23.73%) as a white solid.

LC-MS: m/z＝364.9[M+H] ⁺ (99.30% purity by HPLC, 210nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.75 (d, J＝2.0 Hz, 1H), 7.56-7.50 (m, 2H), 2.69 (s, 3H), 2.58 (s, 3H). (Active hydrogen of sulfonamide is not shown)

Example 2

N ¹ -((5-bromothien-2-yl)sulfonyl)-5-(1-cyanocyclopentyl)-2-hydroxy-N ³ -methylisophthalamide 2

Step 1: 3-Bromo-5-formyl-2-hydroxybenzoic acid 2b

5-Formylsalicylic acid 2a (10.10 g, 60.19 mmol) was dissolved in DMF (100 mL), cooled to 0°C, and N-bromosuccinimide (NBS) (10.87 g, 61.07 mmol) was slowly added thereto. After the addition, the temperature was slowly raised to 70°C and the reaction was continued for 2 hours. TLC monitoring (DCM/MeOH=5/1, _Rf =0.2) showed that the raw material was completely reacted. The reaction solution was quenched with water (300 mL), extracted with ethyl acetate (100 mL x 2), and the organic phases were combined and dried over anhydrous sodium sulfate. The residue was dried and concentrated to give the title compound 2b (15.02 g, crude product) as a yellow solid, which was used directly in the next step.

Step 2 Methyl 3-bromo-5-formyl-2-methoxybenzoate 2c

Compound 2b (15.02 g, crude product) was added to DMF (100 mL), and potassium carbonate (25.34 g, 183.62 mmol) and iodomethane (21.73 g, 153.05 mmol) were added. After the addition was completed, the reaction solution was stirred at room temperature for 18 hours. TLC detection (DCM/MeOH=5/1, R _f =0.2) showed that the raw material reacted completely. The reaction solution was quenched with water (300 mL), extracted with ethyl acetate (100 mL x3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE/EA=10/1) to obtain the title compound 2c (5.40 g, two-step yield 32%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ9.93 (s, 1H), 8.26 (dd, J = 4.8, 2.4Hz, 2H), 4.01 (s, 3H), 3.97 (s, 3H).

Step 3 Methyl 3-bromo-5-(hydroxymethyl)-2-methoxybenzoate 2d

Compound 2c (10.00 g, 36.62 mmol) was added to methanol (200 mL), cooled to 0-5°C, sodium borohydride (813 mg, 21.9 mmol) was slowly added thereto, and stirring was continued for 30 minutes after the addition. TLC detection (PE/EA=4/1, R _f =0.6) showed that the raw material reacted completely, the reaction solution was quenched with water (200 mL), extracted with ethyl acetate (100 mL x3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 2d (10.50 g, crude product) as a yellow oil, which was used directly in the next step.

Step 4: Methyl 3-bromo-2-methoxy-5-(((methylsulfonyl)oxy)methyl)benzoate 2e

Compound 2d (10.50 g, crude product) was added to dichloromethane (100 mL), and triethylamine (5.50 g, 54.4 mmol) was added thereto. The reaction solution was cooled to 0-5°C, and methylsulfonyl chloride (4.68 g, 40.8 mmol) was slowly added dropwise. After the addition, the reaction solution was stirred for 30 minutes. TLC monitoring (PE/EA=4/1, R _f =0.2) showed that the reaction was complete. The reaction solution was quenched by adding saturated brine (100 mL), extracted with dichloromethane (50 mL x2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 2e (11.82 g, crude product) as a yellow oil, which was directly used in the next step.

Step 5: Methyl 3-bromo-5-(cyanomethyl)-2-methoxybenzoate 2f

Compound 2e (11.82 g, crude product) was added to acetonitrile (100 mL), and potassium carbonate (9.24 g, 66.95 mmol) and trimethylsilyl cyanide (5.00 g, 50.39 mmol) were added thereto. After the addition was completed, the reaction solution was stirred at room temperature for 18 hours. TLC detection (PE/EA=4/1, R _f =0.6) showed that the reaction was complete. The reaction solution was quenched with water (100 mL), extracted with ethyl acetate (100 mL x3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE/EA=10/1) to obtain the title compound 2f (4.24 g, three-step yield 40.76%) as a white solid.

Step 6: 2 g methyl 3-bromo-5-(1-cyanocyclopentyl)-2-methoxybenzoate

Compound 2f (104 mg, 0.385 mmol) was added to dry tetrahydrofuran (5 mL), the reaction solution was cooled to 0°C, sodium bis(trimethylsilyl)amide tetrahydrofuran solution (0.45 mL, 0.90 mmol, 2 M/L) was slowly added dropwise, the reaction solution was continued to react at 0°C for 30 minutes, 1,4-dibromobutane (83 mg, 0.39 mmol) was slowly added dropwise, the reaction was continued at 0°C for 30 minutes, and TLC (PE/EA=4/1, R _f =0.3) monitored the reaction to be complete. The reaction solution was quenched by adding saturated brine (20 mL), extracted with dichloromethane (20 mL x3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (PE/EA=5/1) to obtain the title compound 2g (30 mg, yield 24.23%) as a colorless oil.

¹ H NMR (400MHz, CDCl ₃ ) δ7.81(s,2H),3.94(s,3H),3.93(s,3H),2.53-2.46(m,2H),2.08-1.94(m,6H).

Step 7: 3-Bromo-5-(1-cyanocyclopentyl)-2-methoxybenzoic acid 2h

Compound 2g (2.94 g, 8.69 mmol) was added to a mixed solution of methanol (15 mL) and water (5 mL), and sodium hydroxide (452 mg, 11.3 mmol) was added thereto. The reaction solution was reacted at room temperature for 4 hours. TLC monitoring (PE/EA=4/1, R _f =0.6) showed that the reaction was complete. The reaction solution was concentrated under reduced pressure at 20°C to remove the methanol in the reaction solution. Dilute hydrochloric acid (1M/L) was slowly added dropwise to acidify to pH=4-5, and ethyl acetate (20 mL x3) was used for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and the organic phases were concentrated to obtain the title compound 2h (2.81 g, 99%) as a white solid.

LC-MS: m/z=326.0[M+H] ⁺ .

Step 8: 3-Bromo-5-(1-cyanocyclopentyl)-2-methoxy-N-methylbenzamide 2i

Compound 2h (2.81 g, 8.67 mmol) was added to dichloromethane (30 mL), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)urea hexafluorophosphate (6.60 g, 17.3 mmol), methylamine hydrochloride (877 mg, 13.0 mmol) and N,N-diisopropylethylamine (4.47 g, 34.6 mmol) were added thereto. The reaction solution was reacted at room temperature for 18 hours. TLC detection (DCM/MeOH=10/1, R _f =0.2) showed that the reaction was complete. The reaction solution was quenched with water (20 mL), extracted with dichloromethane (50 mL x3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE/EA=2/1) to obtain the title compound 2i (2.62 g, yield 89.73%) as a colorless oily liquid.

Step 9: Methyl 5-(1-cyanocyclopentyl)-2-methoxy-3-(methylcarbamoyl)benzoate 2j

Compound 2i (284 mg, 0.842 mmol) was dissolved in a mixed solution of methanol (10 mL) and DMF (2 mL), and the mixture was transferred into an autoclave. Bistriphenylphosphine palladium dichloride (100 mg, 0.142 mmol) was added thereto. Carbon monoxide was replaced three times, and carbon monoxide was flushed therein. The pressure was maintained at 3-4 atmospheres. The reaction solution was heated to 90°C for 18 hours. TLC monitoring (PE/EA=1/1, _Rf =0.4) showed that a large amount of raw materials remained. The reaction solution was concentrated to dryness, and the crude product was purified by silica gel column chromatography (PE/EA=1/1) to obtain the title compound 2j (21 mg, yield 7.88%) as a colorless oily liquid.

LC-MS: m/z = 317.2 [M + H] ⁺

Step 10: 5-(1-Cyanocyclopentyl)-2-methoxy-3-(methylcarbamoyl)benzoic acid 2k

Compound 2j (61 mg, 0.19 mmol) was added to a mixed solution of methanol (2 mL) and water (2 mL), and sodium hydroxide (15 mg, 0.37 mmol) was added thereto. After addition, the reaction was continued at room temperature for 18 hours. TLC monitoring (PE/EA=1/1, R _f =0.5) showed that the reaction was complete. The methanol in the reaction solution was concentrated and removed, and water (20 mL) was added thereto to quench the reaction. Dilute hydrochloric acid (1 M/L) was slowly added dropwise to acidify to pH=4-5. The mixture was extracted with ethyl acetate (20 mL x3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 2k (30 mg, crude product) as a white solid, which was used directly in the next step.

LC-MS: m/z = 301.1 [MH] ^-

Step 11: N ¹ -((5-bromothiophen-2-yl)sulfonyl)-5-(1-cyanocyclopentyl)-2-methoxy-N ³ -methylisophthalamide 2L

Compound 2k (30 mg, 0.099 mmol) was added to dichloromethane (5 mL), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)urea hexafluorophosphate (60 mg, 0.15 mmol) and N,N-diisopropylethylamine (0.1 mL, 0.5 mmol) were added thereto. After the addition was completed, the reaction solution was reacted at room temperature for 18 hours. TLC monitoring (DCM/MeOH=10/1, R _f =0.2) showed that the reaction was complete. The reaction solution was quenched with water (10 mL), extracted with dichloromethane (5 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was dissolved in dichloromethane (5 mL), and 5-bromothiophene-2-sulfonamide (30 mg, 0.12 mmol) and N,N-diisopropylethylamine (0.1 mL, 0.5 mmol) were added in sequence. The mixture was reacted at room temperature for 4 hours. The reaction was complete under TLC monitoring (DCM/MeOH=20/1, R _f =0.4). The reaction solution was quenched with water (5 mL), extracted with dichloromethane (10 mL x2), and the organic phases were combined and concentrated. The crude product was purified by silica gel column chromatography (DCM/MeOH=60/1) to obtain the title compound 2L (30 mg, two-step yield 30.00%) as a white solid.

LC-MS: m/z = 526.0 [M + H] ⁺

Step 12: N ¹ -((5-bromothiophen-2-yl)sulfonyl)-5-(1-cyanocyclopentyl)-2-hydroxy-N ³ -methylisophthalamide 2

Compound 2L (30 mg, 0.056 mmol) was added to dichloromethane (4 mL), and a boron tribromide-dichloromethane solution (2 mL, 2 mmol, 1 M/L) was added thereto. After the addition was completed, the reaction solution was reacted at room temperature for 1 hour. TLC monitoring (DCM/MeOH=20/1, R _f =0.5) showed that the reaction was complete. The reaction solution was quenched with water (10 mL), extracted with dichloromethane (5 mL x3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/MeOH=20/1) to obtain the title compound 2 (20 mg, yield 68.49%) as a white solid.

LC-MS: m/z=512.0[MH] ⁺ (97.35% purity, 220nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 15.98 (s, 1H), 8.51 (d, J = 4.4 Hz, 1H), 8.02 (dd, J = 6.8, 2.8 Hz, 2H), 7.41 (d, J = 3.6 Hz, 1H), 7.19 (d, J = 4.0 Hz, 1H), 2.83 (d, J = 4.8 Hz, 3H), 2.36 (dd, J = 17.6, 4.8 Hz, 2H), 2.05-1.95 (m, 2H), 1.91-1.84 (m, 4H). (The active hydrogen of sulfonamide is not shown).

Example 3

2,4-Dichloro-N-((2-chloro-4-methylthiazol-5-yl)sulfonyl)benzamide 3

Step 1 2-Chloro-4-methylthiazole-5-sulfonamide 3b

Dissolve 2-chloro-4-methylthiazole-5-sulfonyl chloride 3a (160 mg, 0.689 mmol) in 1,4-dioxane (1 mL), cool to 0°C, slowly add ammonia (0.5 mL), and after addition, warm the reaction solution to room temperature for 10 minutes. TLC shows that the reaction of the raw material is complete. The reaction solution is directly concentrated to obtain the title compound 3b (169 mg, crude product) as a white solid, which is directly used in the next step.

LC-MS: m/z = 212.9 [M + H] ⁺

Step 2 2,4-dichloro-N-((2-chloro-4-methylthiazol-5-yl)sulfonyl)benzamide 3

Compound 3b (213 mg, 1.02 mmol) was dissolved in ethyl acetate (2 mL), triethylamine (322 mg, 3.18 mmol) and 4-dimethylaminopyridine (10 mg, 0.079 mmol) were added in turn at room temperature, followed by a toluene solution (1 mL) of compound IN-1 (214 mg, 1.02 mmol), and the reaction was allowed to proceed overnight at room temperature. TLC showed that the reaction of the raw materials was complete. The reaction solution was quenched with water (2 mL), extracted with ethyl acetate (1 mL x 3), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-TLC (ethyl acetate/petroleum ether = 1/3) to obtain the title compound 3 (60 mg, two-step yield 23%) as a white solid.

LC-MS: m/z＝384.9[M+H] ⁺ (98.43% purity by HPLC, 210nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.64 (d, J=2.0 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.45 (dd, J=8.4, 2.0 Hz, 1H), 2.53 (s, 3H). (The active hydrogen of sulfonamide is not shown).

Example 4

2,4-Dichloro-N-((2-chloro-4-cyclopropylthiazol-5-yl)sulfonyl)benzamide

Step 1-Cyclopropyl-2-thiocyanate-1-one 4b

Compound α-bromocyclopropylethyl ketone 4a (20.1 g, 123 mmol) was dissolved in anhydrous ethanol (170 mL), sodium thiocyanate (12.1 g, 149 mmol) was added at room temperature, and the reaction was continued at room temperature for 10 hours after the addition was completed. The raw material disappeared after TLC monitoring. The filter cake was filtered, washed with anhydrous ether (50 mL x 2), the filtrate was concentrated, the crude product was dissolved with anhydrous ether (50 mL x 2), filtered, and the filtrate was concentrated to obtain the title compound 4b (10.2 g, crude product) as a dark brown liquid, which was directly carried out to the next step.

Step 2 2-Chloro-4-cyclopropylthiazole 4c

Compound 4b (10.2 g, crude product) was dissolved in dichloromethane (120 mL), and dry hydrogen chloride gas was slowly introduced at 0°C for 1 hour, and the temperature was slowly raised to room temperature and the reaction was continued for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (200 mL), extracted with dichloromethane (60 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/5) to obtain the light yellow liquid title compound 4c (4.11 g, two-step yield 20.4%)

LCMS: m/z = 160.1 [M + H] ⁺

¹ H NMR (400MHz, CDCl ₃ ) δ6.73 (s, 1H), 1.96 (tt, J = 8.2, 5.2Hz, 1H), 0.95-0.86 (m, 4H).

Step 3 2-Chloro-4-cyclopropylthiazole-5-sulfonyl chloride 4d

Under nitrogen protection, compound 4c (300 mg, 1.88 mmol) was dissolved in anhydrous tetrahydrofuran (7 mL), cooled to -70 ° C, and n-butyl lithium (2.5 M in n-hexane, 1 mL) was slowly added dropwise, with a slight temperature rise. After addition, the reaction was continued at -70 ° C for 10 minutes. Dry sulfur dioxide gas was introduced for 20 minutes, and the reaction was continued for 10 minutes and then returned to room temperature for 1 hour. The reaction solution was concentrated, the residue was dissolved in dichloromethane (3 mL), N-chlorosuccinimide (376 mg, 2.82 mmol) was added, and the reaction was carried out at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (20 mL), extracted with dichloromethane (10 mL x 3), the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 4d (322 mg, crude product) as a colorless oil, which was directly used in the next step.

Step 4: 2-Chloro-4-cyclopropylthiazole-5-sulfonamide 4e

Compound 4d (322 mg, crude product) was dissolved in 1,4-dioxane (2 mL), and aqueous ammonia (25%, 0.5 mL) was added at room temperature. After addition, the mixture was reacted at room temperature for 10 minutes. TLC showed that the raw material disappeared. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/5) to obtain the title compound 4e (178 mg, two-step yield 40%) as a white solid.

LCMS: m/z = 238.9 [M + H] ⁺

¹ H NMR (400MHz, CDCl ₃ ) δ5.10 (s, 2H), 2.52 (dq, J = 8.0, 5.2Hz, 1H), 1.18-1.08 (m, 4H).

Step 5: 2,4-dichloro-N-((2-chloro-4-cyclopropylthiazol-5-yl)sulfonyl)benzamide 4

2,4-Dichlorobenzoic acid (80 mg, 0.42 mmol) was dissolved in dichloromethane (2 mL), and HATU (239 mg, 0.63 mmol) and N,N-diisopropylethylamine (108 mg, 0.84 mmol) were added at room temperature. After addition, the mixture was reacted at room temperature for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (2 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was dissolved in dichloromethane (2 mL), and compound 4e (100 mg, 0.42 mmol) was added. The mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (2 mL), extracted with dichloromethane (1 mL x 3), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Pre-HPLC (acetonitrile/water = 60/40) to obtain the title compound 4 as a white solid (35 mg, yield 20%)

LC-MS: m/z＝408.9[MH] ^- (99.38% purity by HPLC, 210nm)

¹ H NMR (400 MHz, CDCl ₃ ) δ9.20 (s, 1H), 7.77 (d, J＝8.4 Hz, 1H), 7.48 (d, J＝2.0 Hz, 1H), 7.39 (dd, J＝8.8, 2.0 Hz, 1H), 2.72-2.65 (m, 1H), 1.21-1.13 (m, 4H). (The active hydrogen of sulfonamide is not shown).

Example 5

N-((5-bromothiophen-2-yl)sulfonyl)-2-chloro-4-(trifluoromethyl)benzamide 5

Step 1 2-Chloro-4-(trifluoromethyl)benzoyl chloride 5b

2-Chloro-4-(trifluoromethyl)benzoic acid 5a (202 mg, 0.90 mmol) was added to tetrahydrofuran (10 mL), cooled to 0°C, and N,N-dimethylformamide (4 drops) was added. After the addition, the mixture was reacted at 0°C for 2 minutes, and oxalyl chloride (2 mL, 23.6 mmol) was slowly added dropwise. After the addition, the mixture was heated to room temperature and reacted for 4 hours. TLC showed that the raw material disappeared. The reaction solution was concentrated to dryness to obtain the title compound 5b (230 mg, crude product) as a white solid, which was used directly in the next step.

Step 2 N-((5-bromothiophen-2-yl)sulfonyl)-2-chloro-4-(trifluoromethyl)benzamide 5

5-Bromothiophene-2-sulfonamide (258 mg, 1.06 mmol) was dissolved in dichloromethane (10 mL), and 4-dimethylaminopyridine (5 mg, 0.04 mmol), compound 5b (230 mg, crude product) and triethylamine (2 mL, 14.4 mmol) were added in sequence at room temperature. After addition, the mixture was reacted at room temperature for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-HPLC to obtain the title compound 5 (25 mg, yield 6%) as a white solid.

LC-MS: m/z＝447.8[M+H] ⁺ (96.61% purity by HPLC, 254nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.99 (s, 1H), 7.78 (q, J＝8.4 Hz, 2H), 7.70 (d, J＝4.0 Hz, 1H), 7.42 (d, J＝4.4 Hz, 1H). (Active hydrogen of sulfonamide is not shown)

Example 6

N1-((5-bromothiophen-2-yl)sulfonyl)-4-chloro-2-hydroxy-N3-methylisophthalamide 6

Step 1 4-Chloro-3-formyl-2-hydroxybenzoic acid 6b

4-Chloro-3-formyl-2-hydroxybenzoic acid 6a (3.01 g, 17.4 mmol) was added to trifluoroacetic acid (70 mL), and urotropine (4.86 g, 34.71 mmol) and cuprous oxide (2.48 g, 17.3 mmol) were added at room temperature. After the addition was completed, the reaction solution was heated to 80°C for 10 hours. TLC showed that the raw material disappeared. The reaction solution was cooled to room temperature, and hydrochloric acid (3N, 70 mL) was slowly added dropwise. After the addition was completed, the reaction was continued at room temperature for 2 hours, concentrated, and the residue was added to water (100 mL), filtered, and the filter cake was rinsed with water (300 mL) again and dried to obtain the title compound 6b (7.02 g, crude product) as a white solid, which was directly used in the next step.

Step 2: Methyl 4-chloro-3-formyl-2-hydroxybenzoate 6c

Compound 6b (7.02 g, crude product) was added to anhydrous methanol (70 mL), and concentrated sulfuric acid (7 mL) was added at room temperature. After the addition was completed, the reaction solution was heated to 80 ° C for 10 hours. TLC showed that the raw material disappeared. The reaction solution was cooled to room temperature, quenched with water (100 mL), extracted with ethyl acetate (40 mL x 3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/5) to obtain the title compound 6c as a white solid (3.47 g, two-step yield 93%)

Step 3: Methyl 4-chloro-3-formyl-2-(methoxymethoxy)benzoate 6d

Compound 6c (3.47 g, 16.2 mmol) was added to dichloromethane (100 mL), and triethylamine (3.26 g, 30.3 mmol) was added at room temperature. After the addition was completed, the ice bath was cooled to 0°C, and chloromethyl methyl ether (1.95 g, 24.2 mmol) was added dropwise. After the addition was completed, the mixture was heated to room temperature and stirred for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (100 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/10) to obtain the title compound 6d (2.56 g, yield 61%) as a white solid.

Step 4: 6-Chloro-2-hydroxy-3-(methoxycarbonyl)benzoic acid 6e

Compound 6d (1.00 g, 3.86 mmol) was added to acetonitrile (20 mL), and sodium chlorite (487 mg, 5.41 mmol), sodium dihydrogen phosphate (650 mg, 5.42 mmol) and hydrogen peroxide (3 mL, 30%) were added at room temperature. After addition, the mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. Saturated sodium sulfite aqueous solution (40 mL) was added to the reaction solution, concentrated to a small amount of solvent, and hydrochloric acid (3 N) was slowly added dropwise until pH = 1-2. After stirring for 2 hours, the filtrate was filtered. The cake was dried to give the title compound 6e (500 mg, yield 56%) as a white solid.

Step 5: Methyl 4-chloro-2-hydroxy-3-(methylcarbamoyl)benzoate 6f

Compound 6e (527 mg, 2.28 mmol) was added to dichloromethane (20 mL), and methylamine hydrochloride (246 mg, 3.64 mmol), HATU (1.74 mg, 4.57 mmol) and triethylamine (1 mL, 7.15 mmol) were added at room temperature. After addition, the mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (5 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 6f (450 mg, crude product) as an off-white solid, which was used directly in the next step.

Step 6: 4-chloro-2-hydroxy-3-(methylcarbamoyl)benzoic acid 6g

Compound 6f (310 mg, crude product) was dissolved in tetrahydrofuran (6 mL), and sodium hydroxide aqueous solution (15%, 1 mL) was added at room temperature. After addition, the reaction was continued at room temperature for 10 hours. TLC showed that the starting material disappeared. The reaction solution was quenched with water (6 mL), acidified to pH = 3 with dilute hydrochloric acid (3N), concentrated, filtered, and the filter cake was washed with water (5 mL x 2) and dried to obtain the title compound 6g (190 mg, two-step yield 52%) as a white solid.

Step 7 N ¹ -((5-bromothiophen-2-yl)sulfonyl)-4-chloro-2-hydroxy-N3-methylisophthalamide 6

Compound 6g (190mg, 0.83mmol) was dissolved in dichloromethane (4mL), and N,N-diisopropylethylamine (214mg, 1.65mmol), HATU (472mg, 1.24mmol) and 5-bromo-2-thiophenesulfonamide (220mg, 0.91mmol) were added in sequence at room temperature. After addition, the mixture was reacted at room temperature for 3 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (2mL), extracted with dichloromethane (1mL x3), the organic phases were combined, washed with saturated brine (1mL), and the organic phases were concentrated to obtain the title compound 6 (4.51mg, yield 1.2%) as a white solid.

LC-MS: m/z＝450.9[MH] ^- (97.75% purity by HPLC, 220nm)

¹ H NMR (400 MHz, CDCl ₃ ) δ11.02 (s, 1H), 8.14 (d, J＝8.8 Hz, 1H), 7.84 (s, 1H), 7.71 (d, J＝4.0 Hz, 1H), 7.09 (d, J＝4.0 Hz, 1H), 7.03 (d, J＝8.8 Hz, 1H), 3.09 (d, J＝4.8 Hz, 3H). (Active hydrogen of sulfonamide is not shown)

Example 7

N-((3-bromo-5-methylthiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 7

Step 1 3-Bromo-5-methylthiophene-2-sulfonyl chloride 7b

3-Bromo-5-methylthiophene 7a (500 mg, 2.82 mmol) was dissolved in dichloromethane (5 mL), cooled to 0°C, and a diluent of chlorosulfonic acid (3 mL) in dichloromethane (3 mL) was slowly added. After the addition was completed, the reaction was continued at 0°C for 10 minutes. TLC showed that the raw material disappeared. The reaction solution was slowly poured into ice water (20 mL) for quenching, and extracted with dichloromethane (6 mL x 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 7b (916 mg, crude product) as a colorless oil, which was used directly in the next step.

Step 2 3-Bromo-5-methylthiophene-2-sulfonamide 7c

Compound 7b (916 mg, crude product) was dissolved in 1,4-dioxane (9 mL), and aqueous ammonia (25%, 1 mL) was added at room temperature. After reacting at room temperature for 30 minutes, the starting material disappeared under TLC monitoring. The reaction solution was directly concentrated to obtain the title compound 7c (778 mg, crude product) as a white solid, which was directly used in the next step.

Step 3 N-((3-bromo-5-methylthiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 7

Compound 7c (293 mg, crude product) was dissolved in ethyl acetate (2 mL), triethylamine (386 mg, 3.82 mmol) was added, the mixture was reacted at room temperature for 10 minutes, cooled to 0 ° C, and a mixture of intermediate IN-1 (200 mg, 0.95 mmol) in toluene (1 mL) was added, and the mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (8 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the crude product was purified by Prep-TLC (dichloromethane/methanol = 10/1) to obtain the title compound 7 (110 mg, three-step yield 35%) as a white solid.

LC-MS: m/z＝427.8[MH] ^- (99.88% purity by HPLC, 210nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.73 (d, J＝1.2 Hz, 1H), 7.57-7.50 (m, 2H), 7.10 (s, 1H), 2.52 (s, 3H). (The active hydrogen of sulfonamide is not shown).

Example 8

N-((5-bromothien-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopropyl)-6-methoxybenzamide 8

Step 1: 3-Chloro-4-hydroxy-5-methoxybenzaldehyde 8b

Vanillin 8a (15.1 g, 99.3 mmol) was added to N, N-dimethylformamide (100 mL), and N-chlorosuccinimide (19.9 g, 149 mmol) was added at room temperature. After the addition was completed, the reaction was continued at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (600 mL), extracted with ethyl acetate (200 mL x 3), the organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/15) to obtain the title compound 8b (8.22 g, yield 44%) as a white solid, which was directly used in the next step.

Step 2 3-Chloro-4-trifluoromethanesulfonyl-5-methoxybenzaldehyde 8c

Compound 8b (8.22 g, 44.0 mmol) was added to dichloromethane (150 mL), and triethylamine (13.4 mg, 132 mmol) and 4-dimethylaminopyridine (269 mg, 2.20 mmol) were added at room temperature. After the addition was completed, the reaction solution was cooled to 0 ° C, and N-phenylbis (trifluoromethanesulfonyl) imide solution (20.5 g, 57.3 mmol, dissolved in 40 mL dichloromethane) was added dropwise. After the addition was completed, the temperature was raised to room temperature and reacted for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (200 mL), extracted with dichloromethane (50 mL x 3), the organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 8c (15.3 g, crude product) as an off-white solid, which was directly used in the next step.

Step 3 Methyl 2-chloro-4-formyl-6-methoxybenzoate 8d

Compound 8c (15.3 g, crude product) was added to methanol (465 mL) and N,N-dimethylformamide (20 mL), and triethylamine (19.5 g, 192 mmol), palladium acetate (2.70 g, 12.0 mmol) and 1,1'-bis(diphenylphosphino)ferrocene (2.66 g, 4.81 mmol) were added at room temperature, and carbon monoxide was replaced 3 times. The pressure was maintained at 0.3-0.5 MPa, and the temperature was raised to 80°C for 12 hours. TLC showed that the raw material disappeared. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate (80 mL x 2). The crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/15) to obtain the title compound 8d (8.49 g, two-step yield 85%) as a white solid.

Step 4: Methyl 2-chloro-4-(hydroxymethyl)-6-methoxybenzoate 8e

Compound 8d (8.49 g, 37.1 mmol) was added to methanol (10 mL), and sodium borohydride (702 mg, 18.6 mmol) was added at room temperature. After addition, the mixture was reacted at room temperature for 30 minutes. TLC showed that the starting material disappeared. The reaction solution was quenched with water (80 mL), extracted with ethyl acetate (30 mL x 3), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 8e (6.81 g, crude product) as a light yellow oil, which was used directly in the next step.

Step 5: Methyl 2-chloro-6-methoxy-4-(((methylsulfonyl)oxy)methyl)benzoate 8f

Compound 8e (6.81 g, crude product) was added to dichloromethane (50 mL), cooled to 0°C, triethylamine (6.01 g, 59.1 mmol) was added, and methylsulfonyl chloride (4.06 g, 35.4 mmol) was added after stirring for 10 minutes. After the addition, the mixture was heated to room temperature and reacted for 30 minutes. TLC showed that the raw material disappeared. The reaction solution was quenched with water (100 mL), extracted with dichloromethane (20 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 8f (7.26 g, crude product) as a colorless oil, which was used directly in the next step.

Step 6: 2-chloro-4-(cyanomethyl)-6-methoxybenzoic acid methyl ester 8g

Compound 8f (7.26 g, crude product) was added to acetonitrile (40 mL), and anhydrous potassium carbonate (3.90 g, 28.2 mmol) and trimethylsilyl cyanide (3.50 g, 35.2 mmol) were added in sequence at room temperature. After addition, the temperature was raised to 50 °C and the reaction was continued for 12 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by centrifugation. Purification by silica gel column chromatography (ethyl acetate/petroleum ether=1/5) gave 8 g (1.81 g, three-step yield 20%) of the title compound as a colorless oil.

Step 7: Methyl 2-chloro-4-(1-cyanocyclopropyl)-6-methoxybenzoate 8h

Compound 8g (1.22g, 5.09mmol) was added to tetrahydrofuran (10mL), cooled to 0℃, potassium tert-butoxide (1.26mg, 11.2mmol) was slowly added in batches, reacted at 0℃ for 30 minutes, 1,2-dibromoethane (956mg, 5.09mmol) was added, and the mixture was heated to room temperature for 30 minutes after the addition. TLC showed that the raw material disappeared. The reaction solution was quenched with water (10mL), extracted with ethyl acetate (5mL x3), the organic phases were combined, washed with saturated brine (8mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/5) to obtain the title compound 8h (224mg, yield 16%) as a white solid.

Step 8 2-Chloro-4-(1-cyanocyclopropyl)-6-methoxybenzoic acid 8i

Compound 8h (224 mg, 0.84 mmol) was dissolved in anhydrous methanol (2 mL), and sodium hydroxide solution (15%, 1 mL) and water (0.5 mL) were added at room temperature. After addition, the temperature was raised to 80°C for 10 hours. TLC showed that the starting material disappeared. The reaction solution was cooled to room temperature naturally, acidified to pH = 7 with hydrochloric acid (3N), concentrated to remove methanol, and the residue was diluted with water (1 mL), acidified to pH = 1 with hydrochloric acid (3N). A large amount of white solid precipitated, filtered, and the filter cake was washed with water (1 mL x 2), and dried to obtain the title compound 8i (110 mg, yield 52%) as a white solid.

Step 9: N-((5-bromothiophen-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopropyl)-6-methoxybenzamide 8

Compound 8i (110 mg, 0.44 mmol) was dissolved in dichloromethane (2 mL), and HATU (250 mg, 0.66 mmol), N,N-diisopropylethylamine (113 mg, 0.87 mmol) and 5-bromo-2-thiophenesulfonamide (106 mg, 0.44 mmol) were added in sequence at room temperature. The reaction was carried out at room temperature for 10 hours, and TLC showed that the raw material disappeared. The reaction solution was quenched with water (3 mL), extracted with dichloromethane (1 mL x 3), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and the crude product was purified by Prep-HPLC to obtain the title compound 8 (33 mg, yield 16%) as a white solid.

LC-MS: m/z＝474.9[MH] ^- (98.05% purity by HPLC, 254nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.64 (d, J = 3.6 Hz, 1H), 7.42 (d, J = 4.0 Hz, 1H), 7.03 (d, J = 1.2 Hz, 1H), 6.90 (s, 1H), 3.79 (s, 3H), 1.79 (dd, J = 8.4, 5.2 Hz, 2H), 1.63 (dd, J = 8.0, 4.8 Hz, 2H). (The active hydrogen of sulfonamide is not shown).

Embodiment 9

N-((5-bromothien-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopropyl)-6-hydroxybenzamide 9

Step 1 N-((5-bromothiophen-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopropyl)-6-hydroxybenzamide 9

Compound 8 (20 mg, 42 mmol) was dissolved in dichloromethane (1 mL), and a dichloromethane solution of boron tribromide (1 mL, 1 mmol, 1 N) was added at room temperature. The mixture was reacted at room temperature for 30 minutes. TLC showed that the starting material disappeared. The reaction solution was quenched with water (1 mL), extracted with ethyl acetate (1 mL x 3), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 9 (8.51 mg, yield 43%) as a white solid.

LC-MS: m/z＝488.9[MH] ^- (98.77% purity by HPLC, 220nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.78 (s, 1H), 7.38 (s, 1H), 7.19 (s, 1H), 6.70 (d, J＝2.0 Hz, 1H), 6.68 (s, 1H), 1.74 (dd, J＝8.4, 5.2 Hz, 2H), 1.55 (dd, J＝8.0, 5.2 Hz, 2H). (The active hydrogen of sulfonamide is not shown).

Example 10

(3R,4S,5R)-N-((5-bromothiophen-2-yl)sulfonyl)-3,4,5-trihydroxycyclohexyl-1-ene-1-carboxamide 10

Step 1 (3R,4S,5R)-3,4,5-triacetoxycyclohex-1-ene-1-carboxylic acid 10b

Dissolve (3R,4S,5R)-3,4,5-triacetoxycyclohex-1-ene-1-carboxylic acid 10a (1.02 g, 5.74 mmol) in dichloromethane (10 mL), add triethylamine (1.16 g, 11.5 mmol) at room temperature, react at room temperature for 10 minutes, cool the reaction solution to 0°C and slowly add acetic anhydride (2.34 g, 23.0 mmol), the reaction solution will slightly heat up. After addition, react at room temperature for 30 minutes, TLC shows that the raw material disappears. The reaction solution is quenched with water (10 mL), extracted with dichloromethane (2 mL x 3), the organic phases are combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 10b (1.21 g, crude product) as a colorless oil, which is directly used in the next step.

Step 2 (1R,2S,3R)-5-(Chlorocarbonyl)cyclohex-4-ene-1,2,3-triacetate triester 10c

Compound 10b (1.21 g, crude product) was dissolved in tetrahydrofuran (10 mL), cooled to 0°C, and oxalyl chloride (1.42 g, 11.2 mmol) was slowly added dropwise. After the addition was completed, the mixture was reacted at 0°C for 10 minutes. Then, N,N-dimethylformamide (3 drops) was slowly added dropwise. The reaction was vigorous and a large amount of gas was generated. After the addition was completed, the mixture was heated to 40°C. The mixture was allowed to react at room temperature for 30 minutes, and the reaction was completed by TLC monitoring. The reaction solution was concentrated to obtain the title compound 10c (725 mg, crude product) as a white solid, which was used directly in the next step.

Step 3 (1R,2S,3R)-5-(((5-bromothiophen-2-yl)sulfonyl)carbamoyl)cyclohex-4-ene-1,2,3-triacetate 10d

5-Bromothiophene-2-sulfonamide (826 mg, 3.41 mmol) was dissolved in ethyl acetate (5 mL), triethylamine (1.15 g, 11.4 mmol) and 4-dimethylaminopyridine (28 mg, 0.23 mmol) were added at room temperature, and the mixture was reacted at room temperature for 10 minutes. A solution of compound 10c (725 mg, 2.27 mmol) in toluene (2 mL) was added, and the reaction was continued at room temperature for 12 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (5 mL), extracted with ethyl acetate (2 mL × 3), the organic phases were combined, washed with saturated brine (3 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 10d (1.20 g, crude product) as a colorless oil, which was used directly in the next step.

Step 4: (3R, 4S, 5R)-N-((5-bromothiophen-2-yl)sulfonyl)-3,4,5-trihydroxycyclohexyl-1-ene-1-carboxamide 10

Compound 10d (1.20 g, crude product) was dissolved in anhydrous methanol (10 ml), and anhydrous potassium carbonate (1.26 g, 9.15 mmol) was added at room temperature. After addition, the mixture was reacted at room temperature for 2 hours. TLC showed that the starting material disappeared. The reaction solution was filtered, the filter cake was washed with ethyl acetate (20 mL x 2), the filtrate was concentrated, and the crude product was purified by Prep-HPLC to obtain the title compound 10 (47 mg, four-step yield 2%) as a white solid.

LC-MS: m/z＝397.9[M+H] ⁺ (99.86% purity by HPLC, 254nm)

¹ H NMR (400 MHz, CD ₃ OD) δ7.52 (d, J=4.0 Hz, 1H), 7.11 (d, J=4.4 Hz, 1H), 6.46-6.45 (m, 1H), 4.28 (s, 1H), 3.88 (dt, J=6.7, 4.8 Hz, 1H), 3.60 (dd, J=6.8, 4.0 Hz, 1H), 2.58-2.52 (m, 1H), 2.02 (dd, J=18.0, 4.8 Hz, 1H). (The active hydrogen of sulfonamide is not shown).

Embodiment 11

2,4-Dichloro-N-((5-ethynylthiophen-2-yl)sulfonyl)benzamide 11

Step 1 (Z)-N'-((5-bromothiophen-2-yl)sulfonyl)-N,N-dimethylformimide 11b

5-Bromothiophene-2-sulfonamide 11a (2.40 g, 9.92 mmol) was added to dichloromethane (10 mL), and N,N-dimethylformamide dimethyl acetal (2 mL) was added at room temperature. After addition, the mixture was reacted at room temperature for 20 minutes. TLC showed that the raw material disappeared. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (10 mL x 3), and the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 11b (3.01 g, crude product) as a light yellow solid, which was used directly in the next step.

Step 2 (Z)-N,N-dimethyl-N'-((5-((trimethylsilyl)ethynyl)2-thienyl)sulfonyl)carboximide 11c

Compound 11b (2.00 g, 6.734 mmol) was added to dry N,N-dimethylformamide (20 mL), and triethylamine (4.70 mL, 33.8 mmol), iodide (130 mg, 0.68 mmol), bistriphenylphosphine palladium dichloride (240 mg, 0.34 mmol) and trimethylethynylsilane (1.35 mL, 9.46 mmol) were added in sequence at room temperature. After addition, nitrogen was replaced 3 times, and the reaction was carried out at room temperature for 24 hours. TLC showed that the raw material disappeared. The reaction solution was filtered, the filter cake was washed with ethyl acetate (10 mL x 2), the filtrate was diluted with water (100 mL), and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-TLC (petroleum ether/ethyl acetate = 10/1) to obtain the title compound 11c (1.15 g, two-step yield 37%) as a light yellow solid.

Step 3 5-ethynylthiophene-2-sulfonamide 11d

Compound 11c (1.15 g, 3.66 mmol) was added to dry N,N-dimethylformamide (20 mL), and tetrabutylammonium fluoride solution (7.3 mL, 1 M) was added thereto. After addition, the mixture was reacted at room temperature for 4 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (100 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-TLC (petroleum ether/ethyl acetate = 10/1) to obtain the title compound 11d (450 mg, yield 66%) as a light yellow solid.

Step 4: 2,4-dichloro-N-((5-ethynylthiophen-2-yl)sulfonyl)benzamide 11

2,4-Dichlorobenzoic acid (204 mg, 1.07 mmol) was dissolved in dichloromethane (4 mL), HATU (609 mg, 1.60 mmol) and N,N-diisopropylethylamine (276 mg, 2.14 mmol) were added at room temperature, and the mixture was reacted at room temperature for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (4 mL), extracted with dichloromethane (2 mL x 3), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was dissolved in dichloromethane (4 mL), and N,N-diisopropylethylamine (276 mg, 2.14 mmol) and compound 11d (200 mg, 1.61 mmol) were added in sequence at room temperature. The mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (4 mL) and extracted with dichloromethane (2 mL x 3). The organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 11 (75 mg, yield 19%) as a white solid.

LC-MS: m/z＝359.9[M+H] ⁺ (99.94% purity by HPLC, 254nm)

¹ H NMR (400MHz, CDCl ₃ )δ9.16(s,1H),7.82(d,J＝4.0Hz,1H),7.72(d,J＝8.4Hz,1H),7.44(d,J＝1.6Hz,1H),7.36(dd,J＝8.4,1.6Hz,1H),7.24(d,J＝4.0Hz,1H),3.55(s,1H).

Example 12

2,4-Dichloro-N-((5-iodothien-2-yl)sulfonyl)benzamide 12

Step 1 5-Iodothiophene-2-sulfonyl chloride 12b

2-Iodothiophene 12a (4.01 g, 19.0 mmol) was added to dichloromethane (20 mL), cooled to -60 ° C, and chlorosulfonic acid (5 mL) was slowly added dropwise. After addition, the mixture was reacted at -60 ° C for 10 minutes, and then warmed to room temperature for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched by adding ice water (50 mL), extracted with dichloromethane (20 mL x 3), and the organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 12b (3.02 g, crude product) as a light yellow oil, which was directly used in the next step.

Step 2 5-Iodothiophene-2-sulfonamide 12c

Compound 12b (3.02 g, crude product) was added to 1,4-dioxane (10 mL), and ammonia (2 mL) was added at room temperature. After addition, the mixture was reacted at room temperature for 20 minutes. TLC showed that the starting material disappeared. The reaction solution was poured into water (20 mL) for quenching, extracted with dichloromethane (10 mL x 3), and the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 12c (3.35 g, crude product) as a light yellow oil, which was used directly in the next step.

Step 3 2,4-dichloro-N-((5-iodothien-2-yl)sulfonyl)benzamide 12

Compound 12c (200 mg, crude product) was dissolved in ethyl acetate (1 mL), and triethylamine (2 mL, 14.4 mmol) and toluene solution (1 mL) of intermediate IN-1 (320 mg, crude product) were added successively at room temperature. After addition, the mixture was reacted at room temperature for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (2 mL), extracted with ethyl acetate (1 mL x 3), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 12 (28 mg, three-step yield 5.34%) as a white solid.

LC-MS: m/z＝459.8[MH] ^- (99.89% purity by HPLC, 210nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.72 (d, J=2.0 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 7.51 (dd, J=4.0, 3.2 Hz, 2H), 7.48 (d, J=3.6 Hz, 1H). (The active hydrogen of sulfonamide is not shown).

Example 13

6-Bromo-N-((5-bromothiophen-2-yl)sulfonyl)benzo[b]thiophene-2-carboxamide 13

Step 1 6-Bromo-N-((5-bromothien-2-yl)sulfonyl)benzo[b]thiophene-2-carboxamide 13

5-Bromothiophene-2-sulfonamide 13a (87 mg, 0.36 mmol) was dissolved in N,N-dimethylformamide (1 mL), and N,N-diisopropylethylamine (85 mg, 0.65 mmol), HATU (186 mg, 0.49 mmol) and 6-bromobenzo[B]thiophene-2-carboxylic acid (84 mg, 0.33 mmol) were added in sequence at room temperature. After addition, the mixture was reacted at room temperature for 12 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (2 mL), extracted with ethyl acetate (2 mL x 3), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-TLC (ethyl acetate/petroleum ether = 1/8) to obtain the title compound 13 as a white solid (48 mg, yield 28%).

LCMS: m/z＝477.8[MH] ^- (99.63% purity by HPLC, 254nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 8.22 (d, J=1.6 Hz, 1H), 7.84-7.77 (m, 2H), 7.50 (dd, J=1.6, 2.0 Hz, 1H), 7.33 (d, J=4.0 Hz, 1H), 7.14 (d, J=4.0 Hz, 1H). (The active hydrogen of sulfonamide is not shown).

Embodiment 14

2,4-Dichloro-N-((4,5-dichlorothiophen-2-yl)sulfonyl)benzamide 14

Step 1: 2,4-dichloro-N-((4,5-dichlorothiophen-2-yl)sulfonyl)benzamide 14

The intermediate IN-1 (185 mg, crude product) was dissolved in dichloromethane (10 mL), and 2,3-dichlorothiophene-5-sulfonamide 14a (193 mg, 0.83 mmol) and 4-dimethylaminopyridine (5 mg, 0.05 mmol) were added in sequence at room temperature. After addition, the reaction was allowed to proceed at room temperature for 1 hour. TLC detected that the reaction of the raw materials was complete, and the reaction solution was quenched with water (20 mL), extracted with dichloromethane (10 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 14 (65 mg, yield 19.34%) as a white solid.

LCMS: m/z＝403.8[MH] ^- (99.97% purity by HPLC, 254nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.88 (s, 1H), 7.71 (d, J=2.0 Hz, 1H), 7.61 (d, J=8.4 Hz, 1H), 7.50 (dd, J=8.4, 2.0 Hz, 1H). (The active hydrogen of sulfonamide is not shown).

Embodiment 15

N-((5-bromothiophen-2-yl)sulfonyl l)-6-chloro-2'-cyano-4-hydroxy-[1,1'-biphenyl]-3-carboxamide 15

Step 1 5-Bromo-4-chloro-2-hydroxybenzoic acid 15b

4-Chloro-2-hydroxybenzoic acid 15a (5.40 g, 31.3 mmol) was added to dichloromethane (120 mL), and triethylamine (3.50 g, 34.6 mmol) was added at room temperature. After the addition was completed, the reaction solution was cooled to -60 ° C, and bromine (5.01 g, 31.3 mmol, dissolved in 20 mL dichloromethane) was slowly added dropwise. After the addition was completed, the reaction was continued at -60 ° C for 1 hour. TLC showed that the raw material disappeared. The reaction solution was concentrated, the residue was diluted with water (150 mL), extracted with ethyl acetate (100 mL x 2), the organic phases were combined, washed with dilute hydrochloric acid (1N, 50 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/1) to obtain the title compound 15b as a white solid (2.20 g, yield 28%)

Step 2 Ethyl 5-bromo-4-chloro-2-hydroxybenzoate 15c

Compound 15b (1.00 g, 3.98 mmol) was dissolved in anhydrous ethanol (20 mL), concentrated sulfuric acid (2 mL) was added at room temperature, and the mixture was heated to 80 °C for 10 hours under nitrogen protection. TLC showed that the starting material disappeared. The reaction solution was cooled to room temperature, quenched with water (20 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/10) to obtain the title compound 15c (1.02 g, yield 92%) as a colorless oil.

Step 3 6-Chloro-2'-cyano-4-hydroxy-[1,1'-biphenyl]-3-carboxylic acid ethyl ester 15d

Compound 15c (769 mg, 2.75 mmol) was added to dry N,N-dimethylformamide (20 mL), and 2-cyanophenylboronic acid (566 mg, 3.85 mmol), potassium phosphate (1.16 mg, 5.47 mmol) and Pd(dppf) ₂ Cl ₂ (200 mg, 0.27 mmol) were added in sequence at room temperature. After the addition, the reaction solution was heated to 90°C for 2 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (100 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/8) to obtain the title compound 15d (230 mg, yield 28%) as an off-white solid.

Step 4: 6-Chloro-2'-cyano-4-hydroxy-[1,1'-biphenyl]-3-carboxylic acid 15e

Compound 15d (125 mg, 0.41 mmol) was added to a mixture of ethanol (3 mL) and water (3 mL), and solid sodium hydroxide (50 mg, 1.25 mmol) was added at room temperature. After addition, the reaction solution was reacted at room temperature for 12 hours, and TLC showed that the raw material disappeared. The reaction solution was quenched with water (3 mL), extracted with ethyl acetate (1 mL x 3), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/1) to obtain the title compound 15e (85 mg, yield 76%) as a white solid.

Step 5: N-((5-bromothiophen-2-yl)sulfonyl l)-6-chloro-2'-cyano-4-hydroxy-[1,1'-biphenyl]-3-carboxamide 15

Compound 15e (85 mg, 0.31 mmol) was dissolved in dichloromethane (2 mL), and HATU (177 mg, 0.47 mmol), N,N-diisopropylethylamine (80 mg, 0.62 mmol) and 5-bromo-2-thiophenesulfonamide (75 mg, 0.31 mmol) were added in sequence at room temperature. After addition, the reaction solution was reacted at room temperature for 12 hours. The reaction solution was quenched with water (3 mL), extracted with dichloromethane (1 mL x 3), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-HPLC to obtain the title compound 15 (40 mg, yield 26%) as a white solid.

LC-MS: m/z＝494.9[MH] ^- (98.92% purity by HPLC, 254nm)

¹ H NMR (400 MHz, CDCl ₃ ) δ7.76 (d, J＝7.6 Hz, 1H), 7.70-7.66 (m, 2H), 7.55-7.51 (m, 2H), 7.42 (d, J＝8.0 Hz, 1H), 7.19 (s, 1H), 7.11 (d, J＝3.6 Hz, 1H). (The active hydrogens of sulfonamide and phenolic hydroxyl groups are not shown).

Example 16

N-((5-bromothien-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopentyl)-6-methoxybenzamide 16

Step 1: Methyl 2-chloro-4-(1-cyanocyclopentyl)-6-methoxybenzoate 16a

Compound 8g (1.13g, 4.71mmol) was added to tetrahydrofuran (20mL), cooled to 0℃, and potassium tert-butoxide (1.17g, 10.4mmol) was slowly added in batches. After reacting at 0℃ for 30 minutes, 1,4-dibromobutane (1.02g, 4.73mmol) was added. After the addition was completed, the reaction solution was slowly warmed to room temperature and reacted for 30 minutes. TLC showed that the raw material disappeared. The reaction solution was quenched with water (30mL), extracted with ethyl acetate (15mL x3), the organic phases were combined, washed with saturated brine (10mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/5) to obtain the title compound 16a (1.02g, yield 74%) as a white solid.

Step 2 2-Chloro-4-(1-cyanocyclopentyl)-6-methoxybenzoic acid 16b

Compound 16a (200 mg, 0.68 mmol) was dissolved in anhydrous methanol (6 mL), and sodium hydroxide solution (15%, 1 mL) and water (1 mL) were added at room temperature. After the addition, the reaction solution was heated to 80°C for 10 hours. TLC showed that the raw material disappeared. The reaction solution was naturally cooled to room temperature, and the reaction solution was acidified to pH = 7 by adding dilute hydrochloric acid (3N). The methanol was concentrated to the minimum volume, and the residue was diluted with water (1 mL). Hydrochloric acid (3N) was acidified to pH = 1. A large amount of white solid precipitated, which was filtered, and the filter cake was washed with water (1 mL x 2) and dried to obtain the title compound 16b (165 mg, yield 87%) as a white solid.

Step 3 N-((5-bromothiophen-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopentyl)-6-methoxybenzamide 16

Compound 16b (165 mg, 0.59 mmol) was dissolved in dichloromethane (2 mL), and HATU (336 mg, 0.885 mmol), N,N-diisopropylethylamine (152 mg, 1.18 mmol) and 5-bromo-2-thiophenesulfonamide (143 mg, 0.590 mmol) were added in sequence at room temperature. After addition, the mixture was reacted at room temperature for 10 hours. TLC showed that the raw material disappeared. The reaction solution was quenched with water (3 mL), extracted with dichloromethane (1 mL x 3), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 16 (60 mg, yield 20.19%) as a white solid.

LC-MS: m/z＝501.0[MH] ^- (98.32% purity by HPLC, 220nm)

¹ H NMR (400MHz, CDCl ₃ )δ8.44(s,1H),7.71(d,J＝4.0Hz,1H),7.13(d,J＝4.0Hz,1H),7.04(d,J＝1.2Hz,1 H),6.96(d,J＝1.2Hz,1H),3.86(s,3H),2.47(d,J＝5.2Hz,2H),2.10-1.93(m,6H).

Embodiment 17

N-((5-bromothien-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopentyl)-6-hydroxybenzamide 17

The first step is N-((5-bromothien-2-yl)sulfonyl)-2-chloro-4-(1-cyanocyclopentyl)-6-methoxybenzamide 17

Compound 16 (20 mg, 40 mmol) was dissolved in dichloromethane (1 mL), and a dichloromethane solution of boron tribromide (1 mL, 1 mmol, 1 N) was added at room temperature. After addition, the mixture was reacted at room temperature for 30 minutes. TLC showed that the raw material disappeared. The reaction solution was quenched with water (1 mL), extracted with ethyl acetate (1 mL x 3), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Prep-HPLC to obtain the title compound 17 (10 mg, yield 53%) as a white solid.

LC-MS: m/z＝486.9[MH] ^- (98.77% purity by HPLC, 220nm)

¹ H NMR (400MHz, CDCl ₃ )δ11.77(s,1H),10.04(s,1H),7.74(d,J＝4.0Hz,1H),7.14(d,J＝4.0Hz,1H),7.0 7(d,J＝2.0Hz,1H),7.03(d,J＝2.0Hz,1H),2.50-2.40(m,2H),2.07-1.95(m,6H).

Embodiment 18

2,4-Dichloro-N-((1-ethyl-1H-imidazol-4-yl)sulfonyl)benzamide 18

Step 1-Ethyl-1H-imidazole-4-sulfonyl chloride 18b

1-Ethylimidazole 18a (20.00 g, 208.0 mmol) was slowly added dropwise to chlorosulfonic acid (60 mL), and the temperature of addition was kept below 30°C. After addition, the temperature was raised to 120°C for reaction for 24 hours. The reaction solution was cooled to room temperature, and thionyl chloride (60 mL) was added. After addition, the temperature was raised to 120°C for reaction for 24 hours. TLC detected that the reaction of the raw material was complete (DCM/MeOH=20/1, R _f =0.2). The reaction solution was cooled to 0-10°C. Slowly added dropwise to ice water (200 mL) to quench, extracted with dichloromethane (50 mL x5), combined organic phases, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (EA=100%) to obtain the yellow oily title compound 18b (15.02 g, TLC detected that 2 points could not be separated)

Step 2 1-Ethyl-1H-imidazole-4-sulfonamide 18c

25% to 28% ammonia water (100 mL) was cooled to -15°C, and a tetrahydrofuran solution of compound 18b (15.02 g, crude product, dissolved in 50 mL of tetrahydrofuran) was added dropwise thereto, and the dropping temperature was kept below -10°C. After the dropping was completed, TLC detection (DCM/MeOH=20/1, R _f =0.4) showed that the raw materials reacted completely. The reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (DCM/MeOH=10/1) to obtain 6.52 g of crude product. Ethyl acetate (65 mL) was added and heated to reflux for 30 minutes, and the mixture was naturally cooled to room temperature, filtered, and the solid was washed with ethyl acetate (20 mL). The gray solid was dried to obtain 3.10 g. ¹ H NMR analysis showed that the product contained impurities. The crude product was purified again by silica gel column chromatography (DCM/MeOH=15/1) to obtain 2.01 g. Ethyl acetate (20 mL) was added and heated to reflux for 30 minutes. The mixture was naturally cooled to room temperature, filtered, washed with ethyl acetate (10 mL), and dried to obtain the title compound 18c (1.60 g, two-step yield 4.3%) as an off-white solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.82 (d, J = 0.8Hz, 1H), 7.71 (d, J = 0.4Hz, 1H), 7.14 (s, 2H), 4.03 (q, J = 7.2Hz, 2H), 1.35 (t, J = 7.2Hz, 3H)

Step 3 2,4-dichloro-N-((1-ethyl-1H-imidazol-4-yl)sulfonyl)benzamide 18

2,4-Dichlorobenzoic acid (191 mg, 1.00 mmol) was added to tetrahydrofuran (5 mL), and N,N'-carbonyldiimidazole (370 mg, 2.30 mmol) was added. After the addition, the reaction solution was heated to 60 ° C for 2 hours. The reaction solution was naturally cooled to room temperature and compound 18c (352 mg, 2.00 mmol) was added. The reaction solution was stirred at room temperature for 10 minutes and then 1,8-diazabicyclo[5.4.0]undec-7-ene (300 mg, 1.97 mmol) was added. After the addition, the reaction solution was stirred at room temperature for 18 hours. TLC detection (DCM/MeOH=20/1, R _f =0.2) showed that the reaction was complete. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (20 mL x2), and the aqueous phase was collected. Dilute hydrochloric acid (3 M/L) was slowly added to the aqueous phase to acidify it to pH = 4-5. Ethyl acetate (40 mL x2) was used for extraction, and the organic phases were combined. Dry over anhydrous sodium sulfate, concentrate, add ethyl acetate (20 mL) to the crude product, heat to reflux and slurry for 0.5 h, cool naturally to room temperature, filter, rinse the solid with ethyl acetate (10 mL), and dry to obtain the title compound 18 (250 mg, yield 71.8%) as a white solid.

LC-MS: m/z＝348.0[M+H] ⁺ (98.09% purity, 220nm)

¹ H NMR (400MHz, DMSO-d ₆ )δ12.63(s,1H),8.13(d,J＝1.2Hz,1H),7.94(s,1H),7.72(d,J＝2.0Hz,1H),7.51(dd,J =8.4, 2.0Hz, 1H), 7.45 (d, J = 8.0Hz, 1H), 4.09 (q, J = 7.2Hz, 2H), 1.37 (t, J = 7.2Hz, 3H).

Embodiment 19

N-(1,4-oxazacycline-4-ylsulfonyl)-2,4-dichlorobenzamide 19

Step 1,4-Oxazoline-4-sulfonamide 19b

1,4-Oxazacyclohexane 19a (400 mg, 3.95 mmol) was dissolved in 1,4-dioxane (6 mL), and sulfonamide (456 mg, 4.74 mmol) was added at room temperature. After the addition was completed, the reaction solution was heated to 120°C and reacted for 18 hours. TLC detection (DCM/MeOH=20/1, R _f =0.1) showed that the raw material reacted completely. The reaction solution naturally cooled to room temperature and was concentrated to dryness. Ethyl acetate (10 mL) was added to the residue and heated to reflux until it was completely dissolved. It was filtered while hot, and the filtrate naturally cooled to room temperature. A large amount of solid precipitated. It was filtered, and the filter cake was rinsed with ethyl acetate (10 mL). The filter cake was dried to obtain the yellow solid title compound 19b (250 mg, yield 35.11%).

Step 2 N-(1,4-oxazacycline-4-ylsulfonyl)-2,4-dichlorobenzamide 19

2,4-Dichlorobenzoic acid (300 mg, 1.57 mmol) was added to dichloromethane (10 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (900 mg, 2.36 mmol) and N,N-diisopropylethylamine (1 mL, 5.42 mmol) were added successively at room temperature. After addition, the reaction was continued for 18 hours. TLC detection (DCM/MeOH=20/1, R _f =0.2) showed that the raw material was completely reacted. The reaction solution was quenched with water (10 mL), separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was dissolved in dichloromethane (10 mL), and compound 19b (250 mg, 1.38 mmol) and N,N-diisopropylethylamine (1 mL, 5.42 mmol) were added in sequence at room temperature. After addition, the reaction was continued for 18 hours. TLC detection (DCM/MeOH=20/1, R _f =0.5) showed that the raw material was completely reacted. The reaction solution was quenched with water (20 mL), extracted with dichloromethane (20 mL x2), and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/MeOH=80/1) to obtain the title compound 19 (102 mg, yield 20.9%) as a yellow solid.

LC-MS: m/z＝355.0[M+H] ⁺ (97.44% purity, 220nm)

¹ H NMR (400MHz, DMSO-d ₆ )δ12.19(s,1H),7.76(d,J＝2.0Hz,1H),7.58(d,J＝8.4Hz,1H),7.53(dd,J＝ 8.4,2.0Hz,1H),3.79-3.65(m,4H),3.60-3.53(m,4H),1.92-1.82(m,2H).

Embodiment 20

N-(3-bromo-4-fluorobenzenesulfonyl)-2,4-dichlorobenzamide 20

Step 1 3-Bromo-4-fluorobenzenesulfonamide 20b

3-Bromo-4-fluorobenzenesulfonyl chloride 20a (235 mg, 0.859 mmol) was dissolved in 1,4-dioxane (4 mL), and 25%-28% ammonia water (0.5 mL) was slowly added dropwise. After addition, the mixture was reacted at room temperature for 30 minutes. TLC detection (DCM/MeOH=20/1, R _f =0.8) showed that the raw material was completely reacted. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (15 mL x3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 20b (229 mg, crude product) as a white solid, which was used directly in the next step.

LC-MS: m/z = 251.9 [MH] ^-

Step 2 N-(3-bromo-4-fluorobenzenesulfonyl)-2,4-dichlorobenzamide 20

2,4-Dichlorobenzoic acid (172 mg, 0.900 mmol) was added to dichloromethane (10 mL), and N, N, N', N'-tetramethyl-oxy-(7-azabenzotriazole-1-yl) urea hexafluorophosphate (513 mg, 1.35 mmol) and N, N-diisopropylethylamine (0.2 mL, 1.08 mmol) were added in sequence at room temperature. After the addition, the reaction was continued for 18 hours. TLC detection (DCM/MeOH=20/1, R _f =0.2) showed that the raw material was completely reacted. The reaction solution was extracted with water (10 mL), separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was dissolved in dichloromethane (10 mL), and compound 20b (229 mg, 0.901 mmol) and N, N-diisopropylethylamine (0.2 mL, 1.08 mmol) were added in sequence at room temperature. After the addition, the reaction was continued for 4 hours. TLC detection (DCM/MeOH=20/1, R _f =0.5) showed that the raw material reacted completely. The reaction solution was quenched with water (15 mL), extracted with dichloromethane (10 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/MeOH=30/1) to obtain the title compound 20 (180 mg, two-step yield 49.07%) as a white solid.

LC-MS: m/z＝425.9[M+H] ⁺ (98.97% purity, 210nm)

¹ H NMR (400MHz, CDCl ₃ )δ8.96(s,1H),8.37(dd,J＝6.0,2.4Hz,1H),8.13(ddd,J＝8.8,4.4,2.4Hz,1H),7.72(d,J＝8 .0Hz,1H),7.45(d,J＝4.0Hz,1H),7.36(dd,J＝8.8,2.0Hz,1H),7.31(dd,J＝8.4,8.0Hz,1H).

Embodiment 21

N-((3-bromo-4,5-difluorophenyl)sulfonyl)-2,4-dichlorobenzamide 21

Step 1 4-Bromo-2,3-difluoro-6-nitroaniline 21b

2,3-Difluoro-6-nitroaniline 21a (10.0 g, 57.43 mmol) was dissolved in DMF (230 mL), and N-bromosuccinimide (12.30 g, 69.10 mmol) was slowly added. After the addition, the reaction solution was heated to 90°C and reacted for 5 hours. TLC detection (PE/EA = 10/1, R _f = 0.4) showed that the raw material reacted completely. The reaction solution was cooled to room temperature, quenched with ice water (500 mL), extracted with ethyl acetate (300 mL x 3), and the organic phases were combined. Washed with water (400 mL x 2), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (PE/EA = 7/1) to obtain the yellow solid title compound 21b (13.55 g, yield 93.12%).

¹ H NMR (400MHz, CDCl ₃ ) δ8.23 (dd, J=6.4, 2.4Hz, 1H), 6.25 (s, 2H).

Step 2 1-Bromo-2,3-difluoro-5-nitrobenzene 21c

Compound 21b (12.55 g, 49.60 mmol) was dissolved in tetrahydrofuran (90 mL), and isoamyl nitrite (11.62 g, 98.89 mmol) was slowly added. After the addition, the reaction solution was heated to 55 °C and reacted for 18 hours. TLC detection (PE/EA = 4/1, R _f = 0.5) showed that the raw materials reacted completely, and the reaction solution was cooled to room temperature. Water (100 mL) was added to quench, and ethyl acetate (200 mL x 3) was used for extraction, and the organic phases were combined. Drying with anhydrous sodium sulfate, concentration, and the crude product was purified by silica gel column chromatography (PE = 100%) to obtain the title compound 21c (10.38 g, yield 87.96%) as a yellow oily liquid.

Step 3 1-Bromo-2,3-difluoro-5-nitrobenzene 21d

Compound 21c (10.38 g, 43.61 mmol) was dissolved in ethanol (100 mL), and reduced iron powder (24.42 g, 436.1 mmol), ammonium chloride (23.32 g, 436.0 mmol) and water (50 mL) were added in sequence at room temperature. After the addition, the reaction solution was heated to 80 °C for 18 hours. TLC detection (PE/EA = 4/1, R _f = 0.8) showed that the raw material was completely reacted. The reaction solution was cooled to room temperature, filtered, and the solid was washed with ethanol (100 mL x3). The filtrate was concentrated. The residue was diluted with water (200 mL), extracted with ethyl acetate (200 mL x3), and the organic phases were combined. Drying over anhydrous sodium sulfate, concentration, and the crude product was purified by silica gel column chromatography (PE/EA = 7/1) to obtain the yellow solid title compound 21d (4.80 g, yield 52.9%).

LC-MS: m/z = 210.0 [M + H] ⁺

Step 4: 3-Bromo-4,5-difluorobenzenesulfonyl chloride 21e

Compound 21d (1.01 g, 4.85 mmol) was added to a mixture of concentrated hydrochloric acid (10 mL) and water (10 mL) and stirred evenly. The mixture was cooled to -5°C and a sodium nitrite solution (497 mg, 7.20 mmol, dissolved in 4 mL of water) was slowly added dropwise thereto. The temperature was maintained and the reaction continued for 1 hour. The solution was set aside. Anhydrous copper dichloride (192 mg, 1.42 mmol) was added to acetic acid (15 mL) and sulfur dioxide gas was continuously introduced therein for 1 hour. The mixture was cooled to 5°C. The above diazonium salt solution was added to the reaction and the reaction was carried out at -5°C to 0°C for 2 hours. The reaction was complete as determined by TLC (PE/EA=4/1, R _f =0.3). The reaction solution was quenched by adding ice water (100 mL), extracted with dichloromethane (50 mL x 3), and the organic phases were combined, washed with water (100 mL x 2), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 21e (1.20, crude) as a brown oil, which was used directly in the next step.

Step 5: 3-Bromo-4,5-difluorobenzenesulfonamide 21f

A 25%-28% ammonia solution (10 mL) was cooled to -5°C, and a tetrahydrofuran solution of compound 21e (1.20 g crude product, dissolved in 5 mL tetrahydrofuran) was added dropwise thereto. After the addition was completed, the reaction solution was slowly warmed to room temperature for 30 minutes. TLC detection (PE/EA=4/1, R _f =0.8) showed that the raw material was completely reacted. The reaction solution was quenched by adding saturated saline (50 mL) solution, extracted with ethyl acetate (30 mL x3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/MeOH=40/1) to obtain the title compound 21f (735 mg, two-step yield 55.6%) as a yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.95 (dt, J = 5.6, 2.0Hz, 1H), 7.09-7.85 (m, 1H), 7.66 (s, 2H).

Step 6: N-((3-bromo-4,5-difluorophenyl)sulfonyl)-2,4-dichlorobenzamide 21

2,4-Dichlorobenzoic acid (210 mg, 1.10 mmol) was added to dichloromethane (10 mL), and oxalyl chloride (1 mL, 11.8 mmol) and DMF (1 drop) were added in sequence at room temperature. After the addition was completed, the reaction was carried out at room temperature for 4 hours. TLC detection (DCM/MeOH=20/1, R _f =0.2) showed that the raw material reacted completely. The reaction solution was concentrated to dryness, and the residue was dissolved in dichloromethane (15 mL). Compound 21f (300 mg, 1.10 mmol) and N,N-diisopropylethylamine (0.5 mL, 2.71 mmol) were added in sequence at room temperature. After the addition was completed, the reaction was carried out at room temperature for 2 hours. TLC detection (DCM/MeOH=20/1, R _f =0.5) showed that the raw material reacted completely. The reaction solution was quenched with water (15 mL), extracted with dichloromethane (15 mL x3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give a crude product. Purification by silica gel column chromatography (DCM/MeOH=30/1) gave a white solid (325 mg, HPLC 95%). The crude product was further purified by prep-HPLC to give the title compound 21 as a white solid (200 mg, yield 40%)

LC-MS: m/z＝443.8[MH] ^- (99.91% purity, 210nm)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ8.15-8.02 (m, 2H), 7.74 (d, J＝2.0 Hz, 1H), 7.61 (d, J＝8.0 Hz, 1H), 7.52 (dd, J＝8.4, 2.0 Hz, 1H). (The active hydrogen of sulfonamide is not shown).

Embodiment 22

2,4-Dichloro-N-(morpholinosulfonyl)benzamide 22

Step 1 Morpholine-4-sulfonamide 22b

Morpholine 22a (800 mg, 9.18 mmol) was added to 1,4-dioxane (6 mL), and sulfonamide (1.06 g, 11.0 mmol) was added thereto. After the addition was completed, the reaction solution was heated to 120 °C for 18 hours. TLC monitoring (DCM/MeOH=20/1, R _f =0.2) showed that the reaction was complete, and the reaction solution was cooled to room temperature. The reaction solution was concentrated to dryness, and ethyl acetate (20 mL) was added thereto. After the addition was completed, the reaction solution was heated to reflux until it was completely dissolved. Filter while hot and collect the filtrate. The filtrate naturally cooled to room temperature, and a large amount of solid precipitated. It was filtered, rinsed with ethyl acetate (10 mL), and dried on the filter cake to obtain the yellow solid title compound 22b (805 mg, yield 54.12%).

LC-MS: m/z = 167.1 [M + H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ6.82 (s, 2H), 3.65 (t, J = 4.8Hz, 4H), 2.92 (t, J = 4.8Hz, 4H)

Step 2 2,4-dichloro-N-(morpholinosulfonyl)benzamide 22

2,4-Dichlorobenzoic acid (300 mg, 1.57 mmol) was added to dichloromethane (10 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (900 mg, 2.36 mmol) and N,N-diisopropylethylamine (1 mL, 5.42 mmol) were added thereto. After the addition was completed, the reaction solution was stirred at room temperature for 18 hours. The reaction was completed under TLC monitoring (DCM/MeOH=20/1, R _f =0.2). Water (10 mL) was added thereto for extraction and separation. The mixture was dried over anhydrous sodium sulfate and the organic phase was concentrated to dryness. Dichloromethane (10 mL) was added to dissolve the mixture, and then compound 22b (240 mg, 1.44 mmol) and N,N-diisopropylethylamine (1 mL, 5.42 mmol) were added. After the addition was completed, the mixture was stirred at room temperature for 18 hours. TLC monitoring (DCM/MeOH=20/1, R _f =0.6) showed that the reaction was complete. The reaction solution was quenched with water (10 mL), extracted with dichloromethane (10 mL x2), and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/MeOH=80/1) to obtain the title compound 22 (50 mg, yield 10.25%) as a yellow solid.

LC-MS: m/z＝339.0[M+H] ⁺ (99.82% purity, 254nm)

¹ H NMR (400MHz, DMSO-d ₆ )δ12.25(s,1H),7.77(d,J＝1.6Hz,1H),7.61(d,J＝8.4Hz,1H),7.53(dd,J＝8.4,2.0Hz,1H),3.69-3.62(m,4H),3.32-3.28(m,4H).

Embodiment 23

N-((5-bromo-3-(2-methoxyethyl)thiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 23

Step 1 3-(2-methoxyethyl)thiophene 23b

Under ice bath conditions, sodium hydride (9.36 g, 60%) was added to anhydrous tetrahydrofuran (100 mL), and 3-thiopheneethanol 23a (10.0 g, After the addition, iodomethane (33.0 g, 234 mmol) was slowly added at 0°C. After the addition, the reaction liquid was heated to 60°C under nitrogen protection for 2 hours. TLC monitored that the raw material (ethyl acetate/petroleum ether = 1/10, R _f = 0.2) was completely reacted, and a new point (ethyl acetate/petroleum ether = 1/10, R _f = 0.6) was generated. The reaction liquid was cooled to room temperature and slowly poured into a saturated aqueous ammonium chloride solution (150 mL) at 0°C. It was extracted with ethyl acetate (60 mL x 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the light yellow liquid title compound 23b (13.6 g, crude product), which was directly carried out to the next step.

Step 2 3-(2-Methoxyethyl)thiophene-2-sulfonyl chloride 23c

The dry ice/ethanol bath was cooled to -10°C, and a dichloromethane solution (50 mL) of compound 23b (11.0 g, crude product) was slowly added dropwise to a dichloromethane solution (50 mL) of chlorosulfonic acid (10 mL), (there was a sharp rise in temperature, and the temperature was controlled below 0°C before addition). After the addition was completed, TLC monitored that the raw material (ethyl acetate/petroleum ether = 1/10, R _f = 0.6) disappeared, and a new spot (ethyl acetate/petroleum ether = 1/10, R _f = 0.3) was generated. The reaction solution was slowly poured into ice water (80 mL) for quenching, and extracted with dichloromethane (40 mL x 3). The organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the light yellow liquid title compound 23c (4.46 g, crude product), which was directly carried out to the next step.

Step 3 3-(2-methoxyethyl)thiophene-2-sulfonamide 23d

A tetrahydrofuran solution (20 mL) of compound 23c (4.46 g, crude product) was slowly added dropwise to ammonia water (20 mL), and the reaction was carried out at 20°C for 30 minutes. The raw material (ethyl acetate/petroleum ether = 1/10, R _f = 0.3) disappeared after TLC monitoring. The reaction solution was concentrated, the crude product was diluted with water (50 mL), extracted with ethyl acetate (20 mL x 2), the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/3) to obtain the title compound 23d as a white solid (200 mg, three-step yield 1%)

¹ H NMR (400MHz, CDCl ₃ ) δ7.45 (d, J = 4.8 Hz, 1H), 6.97 (d, J = 5.2 Hz, 1H), 5.39 (s, 2H), 3.66 (t, J = 5.6 Hz, 2H), 3.32 (s, 3H), 3.27 (t, J = 5.6 Hz, 2H).

Step 4: 5-Bromo-3-(2-methoxyethyl)thiophene-2-sulfonamide 23e

Compound 23d (200 mg, 0.90 mmol) was dissolved in dichloromethane (2 mL), and bromine (159 mg, 0.99 mmol) was slowly added at 0°C. After addition, the mixture was reacted at 20°C for 12 hours. The starting material disappeared after TLC monitoring (ethyl acetate/petroleum ether = 1/3, R _f = 0.3). The reaction solution was quenched with water (5 mL), extracted with ethyl acetate (2 mL x 2), the organic phases were combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Pre-TLC (ethyl acetate/petroleum ether = 1/3) to obtain the title compound 23e (40 mg, yield 15%) as a light yellow solid.

LCMS: m/z = 299.9 [M + H] ⁺

¹ H NMR (400MHz, CDCl ₃ ) δ6.95 (s, 1H), 5.39 (s, 2H), 3.63 (t, J = 5.2Hz, 2H), 3.32 (s, 3H), 3.21 (t, J = 5.2Hz, 2H).

Step 5: N-((5-bromo-3-(2-methoxyethyl)thiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 23

2,4-Dichlorobenzoic acid (25 mg, 0.133 mmol) was added to dichloromethane (0.5 mL), and EDCI (33 mg, 0.173 mmol) and HOBT (24 mg, 0.173 mmol) were added in sequence at 20°C. After the addition, the reaction solution was stirred for 1 hour, and the raw material disappeared under TLC monitoring (dichloromethane/methanol = 1/15, R _f = 0.2). The reaction solution was quenched with water (2 mL), extracted with dichloromethane (1 mL x 2), and the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. Compound 23e (40 mg, 0.133 mmol), N,N-diisopropylethylamine (35 mg, 0.266 mmol) and dichloromethane (0.5 mL) were added to the crude product. After addition, the reaction was continued at 20°C for 10 hours. The disappearance of compound 24e (ethyl acetate/petroleum ether = 1/3, R _f = 0.3) was monitored by TLC. The reaction solution was quenched by adding water (2 mL), extracted with dichloromethane (1 mL x 2), and the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Pre-TLC (dichloromethane/methanol = 15/1) to obtain the title compound 23 as a white solid (4 mg, yield 6%)

LCMS: m/z＝471.8[MH] ^- (97.70% purity, 204nm)

¹ H NMR (400MHz, DMSO-d ₆ )δ7.72(s,1H),7.58-7.47(m,2H),7.32(s,1H),7.08(t,J＝51.2Hz,1H),3.56(t,J＝6.4Hz,2H),3.23(s,3H),3.10(t,J＝6.4Hz,2H).

Embodiment 24

5-(2-Acetamidoacetamido)-N-((5-bromothien-2-yl)sulfonyl)-2,4-dichlorobenzamide 24

Step 1: Methyl 5-((tert-Butyloxycarbonyl)amino)-2,4-dichlorobenzoate 24b

Methyl 2,4-dichloro-5-bromobenzoate 24a (12.3 g, 43.3 mmol) was dissolved in toluene (120 mL), and palladium acetate (486 mg, 2.17 mmol), tert-butyl carbamate (6.59 g, 56.3 mmol), BINAP (2.70 g, 4.33 mmol) and cesium carbonate (28.2 g, 86.6 mmol) were added in sequence. After the addition was completed, the atmosphere was replaced with nitrogen three times. The reaction solution was heated to 120°C and reacted for 20 hours. The starting material disappeared under TLC monitoring (ethyl acetate/petroleum ether = 1/5, R _f = 0.5), and a new spot was generated (ethyl acetate/petroleum ether = 1/5, R _f = 0.4). The reaction solution was cooled to room temperature and filtered. The filter cake was washed with ethyl acetate (30 mL x 2). The organic phases were combined and diluted with water (100 mL). The mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/8) to obtain the title compound 24b (6.65 g, yield 47%) as a white solid.

Step 2 5-amino-2,4-dichlorobenzoic acid methyl ester 24c

Compound 24b (6.65 g, 20.8 mmol) was dissolved in dichloromethane (30 mL), trifluoroacetic acid (60 mL) was added at 20°C, and the mixture was kept warm for 1 hour. The starting material disappeared under TLC monitoring (ethyl acetate/petroleum ether = 1/5, R _f = 0.4), and a new spot was generated (ethyl acetate/petroleum ether = 1/5, R _f = 0.2). The reaction solution was concentrated to about 30 mL, diluted with water (50 mL), alkalized to pH = 8-9 by adding 30% sodium hydroxide solution dropwise at 0°C, extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the title compound 24c (4.77 g, crude product) as a white solid, which was directly carried out to the next step.

Step 3 5-(2-acetamidoacetamido)-2,4-dichlorobenzoic acid methyl ester 24d

Compound 24c (4.67 g, crude product) was dissolved in dichloromethane (45 mL), and acetylglycine (2.61 g, 22.3 mmol), HATU (12.1 g, 31.8 mmol) and N,N-diisopropylethylamine (5.48 g, 42.4 mmol) were added in sequence at 20°C. After addition, the reaction solution was reacted at 20°C for 10 hours. TLC monitoring showed that the starting material disappeared (ethyl acetate/petroleum ether = 1/1, R _f = 0.6), and a new spot was generated (ethyl acetate/petroleum ether = 1/1, R _f = 0.2). The reaction solution was quenched with water (70 mL), extracted with dichloromethane (30 mL x 3), the organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 1/1) to obtain the title compound 24d (11.2 g, crude product) as a light yellow solid, which was directly used in the next step.

LCMS: m/z = 319.0 [M + H] ⁺

Step 4: 5-(2-acetamidoacetamido)-2,4-dichlorobenzoic acid 24e

Compound 24d (10.7 g, crude product) was dissolved in anhydrous methanol (100 mL), and solid sodium hydroxide (1.34 g, 33.5 mmol) and water (100 mL) were added at 10°C. After addition, the reaction was continued at 20°C for 3 hours. The starting material disappeared under TLC monitoring (dichloromethane/methanol = 10/1, R _f = 0.7), and a new spot was generated (dichloromethane/methanol = 5/1, R _f = 0.1). The reaction solution was concentrated, and hydrochloric acid (25 mL, 6N) was added at 0°C to acidify to pH = 2-3, and ethyl acetate (40 mL x 4) was used for extraction. The organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, concentrated, slurried at room temperature (ethyl acetate/petroleum ether = 1/3, 50 mL), filtered, and the filter cake was dried to obtain the title compound 24e (7.46 g, three-step yield 73%) as a light yellow solid.

Step 5: 5-(2-acetamidoacetamido)-N-((5-bromothiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 24

Compound 24e (300 mg, 0.98 mmol) was added to dichloromethane (3 mL), and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (767 mg, 1.47 mmol) and N,N-diisopropylethylamine (254 mg, 1.97 mmol) were added in sequence at room temperature. After addition, the reaction was continued for 2 hours. The starting material disappeared under TLC monitoring (dichloromethane/methanol = 5/1, R _f = 0.1). The reaction solution was quenched with water (5 mL), extracted with dichloromethane (1 mL x 2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. 5-bromo-2-thiophenesulfonamide (262 mg, 1.08 mmol), N,N-diisopropylethylamine (254 mg, 1.966 mmol) and dichloromethane (3 mL) were added to the crude product. After addition, the reaction was continued at room temperature for 10 hours. The active ester of compound 25e disappeared under LCMS monitoring. The reaction solution was quenched with water (5 mL), extracted with dichloromethane (2 mL), and the organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by pre-TLC (dichloromethane/methanol=10/1) to give the title compound 24 (120 mg, yield 23%) as a light yellow solid.

LCMS: m/z＝527.8[M+H] ⁺ (99.78% purity, 210nm)

¹ H NMR (400MHz, DMSO-d ₆ )δ9.67(s,1H),8.30(t,J＝5.6Hz,1H),7.95(s,1H),7.77(s,1H),7.65(d,J＝4.0Hz,1H ),7.39(d,J＝4.1Hz,1H),7.10(t,J＝50.8Hz,1H),3.94(d,J＝6.0Hz,2H),1.89(s,3H).

Embodiment 25

5-(2-acetamidoacetamido)-N-((5-bromo-3-(2-methoxyethyl)thiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 25

Step 1 5-(2-acetamidoacetamido)-N-((5-bromothiophen-2-yl)sulfonyl)-2,4-dichlorobenzamide 25

Compound 24e (97 mg, 0.32 mmol) was added to dichloromethane (1 mL), and EDCI (80 mg, 0.41 mmol) and HOBT (56 mg, 0.0.41 mmol) were added in sequence at 20°C. After the addition, the reaction solution was stirred for 1 hour, and the starting material disappeared under TLC monitoring (dichloromethane/methanol = 1/15, R _f = 0.2). The reaction solution was quenched with water (2 mL), extracted with dichloromethane (1 mL x 2), and the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was added with compound 23e (100 mg, 0.33 mmol), N,N-diisopropylethylamine (205 mg, 1.58 mmol) and dichloromethane (1 mL). After the addition was completed, the reaction was continued at 20°C for 10 hours. The disappearance of compound 24e (ethyl acetate/petroleum ether = 1/3, R _f = 0.3) was monitored by TLC. The reaction solution was quenched with water (2 mL), extracted with dichloromethane (1 mL x 2), and the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by Pre-TLC (dichloromethane/methanol = 1/10) to obtain the title compound 25 as a white solid (20 mg, yield 11%)

LCMS: m/z＝585.9[M+H] ⁺ (95.27% purity, 204nm)

¹ H NMR (400MHz, DMSO-d ₆ )δ9.67(s,1H),8.31(t,J＝5.6Hz,1H),7.95(s,1H),7.77(s,1H),7.31(s,1H),7.09(t,J＝51.2Hz,1 H), 3.94 (d, J = 6.0Hz, 2H), 3.55 (t, J = 6.8Hz, 2H), 3.22 (s, 3H), 3.09 (t, J = 6.8Hz, 2H), 1.89 (s, 3H).

Embodiment 26

N-((5-bromothiazol-2-yl)sulfonyl)-2,4-dichlorobenzamide 26

Step 1 2-(Benzylthio)-5-bromothiazole 26b

Benzyl mercaptan (6.14 g, 49.40 mmol) was added to a DMF (50 mL) solution of 2,5-dibromothiazole 26a (10.0 g, 41.17 mmol) and potassium carbonate (11.38 g, 82.33 mmol), and stirred at 25°C for 16 hours. TLC showed that the reaction was complete (PE/EA=15/1, raw material R _f =0.5, product R _f =0.55). The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (50 mL), washed with water (50 mL x3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was separated and purified by silica gel column chromatography to obtain the light yellow liquid title compound 26b (10.6 g, yield 90.0%).

Step 2 5-Bromothiazole-2-sulfonyl chloride 26c

At 0°C, N-chlorosuccinimide (9.42 g, 70.58 mmol) was added to a mixed solution of compound 26b (10.1 g, 35.29 mmol) in acetic acid/water (3:1, 200 mL). The reaction solution was heated to 25°C and stirred for 2 hours. TLC showed that the reaction was complete (PE/EA = 10/1, raw material R _f = 0.5, product R _f = 0.3). The reaction solution was quenched with water (150 mL), extracted with ethyl acetate (150 mL), washed with water (150 mL x 2), washed with saturated sodium bicarbonate solution (150 mL x 2), washed with saturated brine (150 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography. The title compound 26c (8.75 g, crude product) was obtained as a light yellow liquid and was used directly in the next step.

Step 3 5-Bromothiazole-2-sulfonyl chloride 26d

At 0°C, ammonia water (5 mL) was added to a tetrahydrofuran (50 mL) solution of compound 26c (8.75 g, crude product), and the reaction solution was heated to 25°C and stirred for 30 minutes. TLC showed that the reaction was complete (PE/EA=3/1, raw material R _f =0.5, product R _f =0.3). The reaction solution was concentrated, acidified to pH=3-4 with 2M hydrochloric acid, extracted with ethyl acetate (50 mL), washed with water (5 mL x2), washed with saturated brine (50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was separated and purified by silica gel column chromatography to obtain the title compound 26d (2.76 g, two-step yield 32.2%) as a white solid.

LC-MS: m/z = 242.9 [M + H] ⁺

Step 4: N-((5-bromothiazol-2-yl)sulfonyl)-2,4-dichlorobenzamide 26

At 0°C, oxalyl chloride (206 mg, 1.62 mmol) and DMF (1 drop) were added to a suspension of 2,4-dichlorobenzoic acid (155 mg, 0.81 mmol) in DCM (2 mL). The reaction mixture was heated to 25°C for 1 hour. The reaction mixture was concentrated and the concentrate was dissolved in DCM (2 mL). At 0°C, the DCM solution of the concentrate was added to DCM (3 mL) of compound 26d (150 mg, 0.62 mmol) and triethylamine (187 mg, 1.85 mmol), and the mixture was heated to 25°C and stirred for 30 minutes. TLC showed that the reaction was complete (PE/EA=2/1, raw material R _f =0.3, DCM/MeOH=10:1, product R _f =0.1). The reaction solution was concentrated, and the crude product was dissolved in water (15 mL), extracted with PE/EA (3/1, 15 mL x3), the aqueous phase was acidified to pH=3-4 with 2M hydrochloric acid, extracted with ethyl acetate (15 mL), and water (5 mL x2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was suspended in DCM (5 mL), PE (15 mL) was added and stirred for 15 minutes, filtered, and the filter cake was washed with PE/EA (3/1, 2 mL), drained, and dried to obtain the title compound 26 (128 mg, yield 57.6%) as a white solid.

LC-MS: m/z＝414.8[M+H] ⁺ (97.46% purity by HPLC, 254nm)

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.24 (s, 1H), 7.72 (d, J = 1.6 Hz, 1H), 7.59 (d, J = 8.0 Hz, 1H), 7.51 (dd, J = 8.4, 2.0 Hz, 1H).

Embodiment 27

2,4-Dichloro-N-((5-ethynylthiazol-2-yl)sulfonyl)benzamide 27

Step 1 5-((Trimethylsilyl)ethynyl)thiazole-2-sulfonamide 27a

Cuprous iodide CuI (20 mg, 0.1 mmol) and Pd(PPh ₃ ) ₂ Cl ₂ (72 mg, 0.1 mol) were added to a DMF (5 mL) solution of compound 26d (500 mg, 2.06 mmol) and triethylamine (624 mg, 6.17 mmol). The atmosphere was replaced with nitrogen three times for protection. Trimethylethynylsilane (1.14 g, 11.57 mmol) was added into the reaction solution by syringe. The reaction solution was heated to 80° C. and stirred for 30 minutes. TLC showed that the reaction was completed (PE/EA=1/1, raw material R _f =0.3, product R _f =0.35). EA (50 mL) was added to the reaction solution, solid precipitated, and the solid was filtered. The filter cake was washed with EA (5 mL), and the filtrate was washed with water (50 mL x 3). The EA phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified by silica gel column chromatography to obtain the title compound 27a (310 mg, 57.9%) as a light yellow liquid.

LC-MS: m/z = 261.0 [M + H] ⁺

Step 2 5-ethynylthiazole-2-sulfonamide 27b

Potassium carbonate (119 mg, 0.86 mmol) was added to methanol (5 mL) of compound 27a (150 mg, 0.57 mmol) at 25°C, and the reaction solution was stirred for 30 minutes. TLC showed that the reaction was complete (PE/EA=5/1, raw material R _f =0.5, product R _f =0.3). The reaction solution was diluted with water (10 mL), acidified to pH=3 with 2M hydrochloric acid, extracted with ethyl acetate (15 mL), washed with water (15 mL x2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the light yellow liquid title compound 27b (108 mg, 100%).

LC-MS: m/z = 189.0 [M + H] ⁺

Step 3 2,4-dichloro-N-((5-ethynylthiazol-2-yl)sulfonyl)benzamide 27

At 0°C, oxalyl chloride (174 mg, 1.37 mmol) and DMF (1 drop) were added to a suspension of 2,4-dichlorobenzoic acid (131 mg, 0.69 mmol) in DCM (2 mL). The reaction solution was heated to 25°C for 1 hour. The reaction solution was concentrated and the concentrate was dissolved in DCM (2 mL). At 0°C, the above solution was added to a suspension of compound 27b (108 mg, 0.57 mmol) and triethylamine (174 mg, 1.72 mmol) in DCM (3 mL). The reaction solution was heated to 25°C and stirred for 30 minutes. TLC showed that the reaction was complete (PE/EA=2/1, raw material R _f =0.3, DCM/MeOH=10:1, product R _f = 0.1), the reaction solution was concentrated, the crude product was dissolved in water (15 mL), extracted with PE/EA mixed solution (3/1, 15 mL x3), the aqueous phase was acidified with 2M hydrochloric acid to pH = 3-4, extracted with ethyl acetate (15 mL), washed with water (5 mL x2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was suspended in DCM (5 mL), petroleum ether (15 mL) was added and stirred for 15 minutes, filtered, and the filter cake was washed with PE/EA (3/1, 2 mL) and dried to obtain the title compound 27 (118 mg, yield 56.9%) as a white solid.

LC-MS: m/z＝360.9[M+H] ⁺ (99.98% purity by HPLC, 254nm)

¹ H NMR (400MHz, DMSO-d ₆ ) δ8.34(s,1H),7.71(d,J＝2.0Hz,1H),7.59(d,J＝8.4Hz,1H),7.51(dd,J＝8.0,2.0Hz,1H),5.10(s,1H).

Pharmacological activity test

Test Example 1: Affinity test of compounds with WDR5 at the molecular level

The affinity test of the compounds in the present disclosure to WDR5 at the molecular level was carried out by the following method:

Compound preparation: Weigh the compound accurately and dissolve it in DMSO (Sigma, D5879) to 10mM stock solution for use. Use 1x HBS-EP buffer (10mM HEPES, pH 7.4, 150mM NaCl, 3.0mM EDTA, and 0.005% (v/v) TW-20) to dilute the stock solution 2-fold to 9 concentrations. The final concentration of the compound in the reaction system is 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78, 0.39μM, and the final DMSO concentration of the compound is 5%. According to the corresponding experimental procedure design, a certain amount of compound samples are transferred to a 96-well plate (Greiner, 650101) in sequence.

Surface plasmon resonance experiment (SPR): WDR5 protein was coupled to a CM5 chip (Cytiva, BR100530) using the amino coupling method, with a signal value of approximately 9000 response units (RU). Three initialization cycles were first run, followed by sequential injection of the test compounds, with the binding time set to 120 seconds and the dissociation time set to 100 seconds, followed by washing the residual compounds in the chip with a solution containing 50% DMSO. Solvent correction was performed every 48 cycles using 1x HBS-EP buffer containing 4.5%-5.8% DMSO.

Detection and analysis: Biacore8K was used for signal detection, data collection, processing and analysis. The raw data generated by the experiment was processed by Biacore8K data processing software by subtracting the control group parameters and solvent correction, and the static affinity model was used to fit the corresponding _KD value, as shown in Table 1.

Conclusion: The compounds of the embodiments of the present disclosure have a strong affinity with the WDR5 protein at the molecular level.

Table 1. Affinity test results of the compounds disclosed in the present invention and WDR5 protein

Test Example 2: Anti-proliferation activity test of compounds on neuroblastoma cells

The antiproliferative activity of the compounds of the present disclosure on neuroblastoma cells was tested by the following method:

Compound preparation: Weigh the compound accurately and dissolve it in DMSO to 10mM stock solution for later use. Use MEM or RPMI 1640 medium to dilute the compound to a final test concentration of 10μM and a final DMSO concentration of 0.1%, and transfer the compound to a 96-well plate for subsequent experiments.

Cell Counting Kit-8 Test (CCK-8): 293T, SK-N-AS, IMR32 and LAN5 cells were inoculated into 96-well cell culture plates at a seeding volume of 2x10 ⁴ cells/well. The corresponding compounds and DMSO control were added to each well and cultured for 72 hours. Then 10 μL of CCK-8 reagent was added to each well and incubated at 37 degrees for 2 hours. The absorbance of the cells at 450 nm was measured using a microplate reader.

Detection and analysis: Collect the absorbance data corresponding to the cells in each well, and calculate the corresponding cell survival rate and inhibition rate, see Table 2 for details.

Table 2. Antiproliferative activity test results of some compounds in the present disclosure on neuroblastoma cells

Conclusion: The compounds of the examples disclosed in this disclosure have good anti-proliferative activity against neuroblastoma cells. Wherein "-" indicates that the activity was not tested.

Test Example 3: Structural analysis of the complex of some compounds disclosed herein and WDR5

The structural elucidation of the complex of selected compound 13 and compound 14 in the present disclosure was performed by the following method:

Protein crystal preparation: The expression vector Rosetta2 competent cells of WDR5 were placed in a 37°C constant temperature shaking incubator and cultured until the OD value was 0.6-0.8. IPTG with a final concentration of 0.1mM was added and cultured at 16°C for 20 hours, and the cells were collected. The cells were broken using a high-pressure cell disruptor and the supernatant was obtained after high-speed centrifugation for 30 minutes. 10mL of Ni-NTA affinity chromatography gel resin was added, stirred and bound at 4°C for 1 hour, and the supernatant was obtained by centrifugation. The resin was treated with impurity removal buffer (50mM Tris pH 7.5, 150mM NaCl, 50mM imidazole, 1mM TCEP) and elution buffer (50mM Tris pH 7.5, 150mM NaCl, 300mM imidazole, 1mM TCEP) to obtain the preliminary target protein. Afterwards, an anion exchange chromatography column and a molecular exclusion chromatography column were used to obtain high-purity WDR5 protein, and samples were collected and protein purity was identified using SDS-polyacrylamide gel electrophoresis.

The obtained high-purity WDR5 protein was concentrated to 11 mg/mL using an ultrafiltration tube and centrifuged at 12000 rpm for 30 minutes. Crystallization buffer was prepared: 0.1 M Bis-Tris, pH 5.8, 0.2 M ammonium acetate, 30% (w/v) pEG3350. Crystallization conditions were optimized using a 24-well hanging drop plate. 1 mL of buffer per well. The two solutions were mixed in a ratio of 1.5 μL crystallization buffer + 1.5 μL protein solution, and crystals were grown in a constant temperature incubator at 18°C for 2-3 days. The crystals were in the form of flakes. The complex crystals of WDR5 and active compounds were obtained by immersion method.

Detection and analysis: The diffraction data of the crystals were collected at the microcrystal complex beamline BL18U1 of the Shanghai Synchrotron Radiation Source (SSRF). The collected data were processed using HKL3000, including data point selection, indexing, correction, integration, data merging and normalization, to generate MTZ files. The WDR5 empty protein crystal structure resolved by this laboratory was used as a template, and the Phaser module in CCP4 was used for molecular replacement to generate the initial structural coordinates. COOT was used for model construction and optimization, and then Phenix was used to further optimize the model, and the R factor in the structural parameters was repeatedly made less than 0.25. The results are shown in Figure 1.

Conclusion: The compounds of the examples disclosed herein bind to WDR5 by occupying the WBM site to block the interaction between WDR5 and Myc.

Test Example 4: In vitro efficacy test of some compounds disclosed herein on acute myeloid leukemia, hepatoma cell lines and pancreatic cancer cell lines

Compound preparation: Weigh the compound accurately and dissolve it in DMSO to a 10mM stock solution for later use. Use MEM or RPMI 1640 medium to dilute the compound to a final test concentration of 10μM and a final DMSO concentration of 0.1%, and transfer the compound to a 96-well plate for subsequent experiments.

Cell Counting Kit-8 Test (CCK-8): Hep3B, Huh-7, MV4-11 and Mia-Paca2 cells were inoculated into 96-well cell culture plates at a seeding volume of 2x10 ⁴ cells/well. The corresponding compounds and DMSO control were added to each well and cultured for 72 hours. Then 10 μL of CCK-8 reagent was added to each well and incubated at 37 degrees for 2 hours. The absorbance of the cells at 450 nm was measured using a microplate reader.

Detection and analysis: Collect the absorbance data corresponding to the cells in each well and calculate the corresponding inhibition rate, see Table 3 for details.

Table 3. In vitro efficacy test results of some compounds of the present disclosure on acute myeloid leukemia, hepatoma cell lines and pancreatic cancer cell lines

Conclusion: The disclosed example compounds have good anti-proliferative activity against hepatoma cells, acute myeloid leukemia cells and pancreatic cancer cells.

Test Example 5: Pharmacokinetic test of some compounds disclosed in the present invention on normal mice

This test example studied the pharmacokinetic properties of compound 11 in mice. Balb/c female mice, 6-8 weeks old, were selected and injected with compound 11 by gavage or tail vein. Whole blood was collected at 5min, 15min, 30min, 1h, 2h, 4h, 8h, and 24h after administration. After centrifugation at 8000rpm for 5min, the upper plasma was collected. The plasma exposure of the drug at different time points was tested by HPLC-MS/MS method, and the pharmacokinetic parameters were calculated. The specific experimental results are as follows:

Table 4. Pharmacokinetic test results of some compounds of the present disclosure on normal mice

Conclusion: The compound of the embodiment of the present disclosure is rapidly absorbed orally, has extremely high bioavailability, and has a plasma half-life T _1/2 of up to 15.9 h, making it an ideal potential oral drug candidate molecule.

Test Example 6: Efficacy test of some compounds disclosed herein on the IMR32 CDX mouse neuroblastoma model

The efficacy of compound 11 on the neuroblastoma model was studied. Human neuroblastoma IMR32 cells were selected and cultured in vitro in a monolayer. The cells were routinely digested and passaged with trypsin-EDTA twice a week. When the cell saturation was 80%-90%, the cells were collected, counted, and the inoculation concentration was adjusted to 10 million/mouse. The cells were inoculated subcutaneously on the upper right back of the animal. When the tumor grew to about 120 mm ³ , the animals were randomly grouped and dosed according to their weight and tumor volume. The weight and tumor volume of the animals were measured twice a week. The efficacy data were expressed in terms of tumor volume (TV), tumor inhibition rate (TGI), and relative volume (T/C). The specific experimental data are as follows:

Table 5. Results of efficacy test of some compounds disclosed in the present invention on IMR32 CDX mouse neuroblastoma model n=6

The experimental results showed that there was no obvious abnormality in the health status of the animals during the administration period; compared with the solvent control (Vehicle) group, the low, medium and high dose groups of the disclosed embodiment compound all showed obvious anti-tumor efficacy and had a good dose-effect relationship, with TGI reaching 85.5%, 107.1% and 107.1% respectively, and the tumors of some animals in the high and medium dose groups completely disappeared in the late stage of administration. It can be seen that the disclosed embodiment compound has significant efficacy in the IMR32 human neuroblastoma model.

Test Example 7: Efficacy test of some compounds disclosed herein on Hep3B CDX mouse hepatoma model

This test example studied the efficacy of compound 11 on a liver cancer model. Human liver cancer Hep3B cells were selected for in vitro monolayer culture and routine digestion and passage using trypsin-EDTA twice a week. When the cell saturation was 80%-90%, the cells were collected, counted, and the inoculation concentration was adjusted to 10 million cells/mouse. The cells were inoculated subcutaneously on the upper right back of the animal. When the tumor grew to about 110 mm ³ , the animals were randomly grouped and dosed according to their weight and tumor volume. The animal weight and tumor volume were measured twice a week. The efficacy data were expressed in terms of tumor volume (TV), tumor inhibition rate (TGI), and relative volume (T/C). The specific experimental data are as follows:

Table 6. Results of efficacy test of some compounds disclosed in the present invention on Hep3B CDX mouse hepatoma model n=5

The experimental results showed that there was no obvious abnormality in the health status of the animals during the administration period; compared with the solvent control (Vehicle) group, the low-dose and high-dose groups of the disclosed embodiment compounds showed obvious anti-tumor efficacy and good dose-effect relationship, with TGI reaching 40% and 106.8% respectively, and the tumors of some animals in the high-dose 90 mg/kg group completely disappeared in the late stage of administration. It can be seen that the disclosed embodiment compounds have significant efficacy in the Hep3B liver cancer xenograft tumor model.

Test Example 8: Safety test of some compounds disclosed in the present invention on normal mice

This test example studied the safety of compound 11. Female ICR mice aged 6-8 weeks were selected as the research subjects. The animals were randomly divided into groups according to their weight and orally administered at 100 mg/kg, 300 mg/kg, 400 mg/kg, and 500 mg/kg, respectively. The weight changes, morbidity, and mortality of the animals were observed. The specific results are as follows:

Table 7. Safety test results of some compounds of the present disclosure on normal mice

The experimental results show that, in ICR mice, oral administration of the disclosed embodiment compound at doses of 100 mg/kg, 300 mg/kg and 400 mg/kg did not cause disease or death in all animals, and only one animal in the 500 mg/kg group died on the third day of the experiment. It can be seen that the expected tolerable dose of the disclosed embodiment compound is as high as 400 mg/kg, and the safety is good.

Test Example 9: Protein Binding Rate Test of Some Compounds Disclosed

The plasma protein binding rate of compound 11 in human plasma was determined by equilibrium dialysis. The left and right chambers were separated by a semipermeable membrane. The drug-containing protein solution was added to the left side, and the blank buffer was added to the right side. The unbound free drug can freely pass through the semipermeable membrane. After a certain period of incubation, the two sides reached equilibrium, and the free drug concentration was equal. The plasma protein binding rate can be calculated by measuring the drug concentration on both sides. The results are shown in Table 8:

Table 8. Protein binding test results of some compounds disclosed in this disclosure

The experimental results show that the free drug content of the compound of the embodiment of the present disclosure reaches 1.48%, which is sufficient to meet the therapeutic drug concentration requirement in vivo.

Test Example 10: Hepatocyte Stability Test of Some Compounds of the Disclosure

Transfer 198 μL of live cell suspension to a 96-well deep-well plate, place the deep-well plate on a vortex and preheat in an incubator for 10 minutes. The experiment was performed in parallel. Add 2 μL of 100 μM test substance or positive drug verapamil to each well to start the reaction, and place the deep-well plate back on the incubator vortexer. Incubate the sample, take 25 μL of the suspension at 0, 15, 30, 60, 90 and 120 minutes, and add 150 μL of acetonitrile containing the internal standard to terminate the reaction. Vortex for 10 minutes and centrifuge at 3220g and 4°C for 45 minutes. Transfer 100 μL of supernatant to the sample injection plate, add 100 μL of pure water to mix, and use for UPLC-MS/MS analysis. The results are shown in Table 9:

Table 9. Results of the hepatocyte stability test of some compounds of the present disclosure

The experimental results show that the compounds of the embodiments of the present disclosure are easily metabolized and eliminated in the human liver and are not prone to cumulative toxicity.

The applicant declares that the present disclosure uses the above-mentioned embodiments to illustrate the sulfonamide compounds disclosed herein as WDR5 and Myc interaction inhibitors and their preparation methods and applications, but the present disclosure is not limited to the above-mentioned embodiments, that is, it does not mean that the present disclosure must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvements to the present disclosure, equivalent replacement of the raw materials of the products disclosed herein, addition of auxiliary components, selection of specific methods, etc., all fall within the scope of protection and disclosure of the present disclosure. The above describes the present disclosure in detail. The optional implementation modes disclosed herein, however, are not limited to the specific details in the above implementation modes. Within the technical concept of the present disclosure, the technical solution of the present disclosure can be subjected to a variety of simple modifications, and these simple modifications all belong to the protection scope of the present disclosure. It should also be noted that the various specific technical features described in the above specific implementation modes can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present disclosure will not further describe various possible combinations.

Claims (8)

A sulfonamide compound represented by formula (I) or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof,
in: ^A1 ring and ^A2 ring are each independently selected from one of a C6-C30 aryl ring, a C6-C30 aliphatic hydrocarbon ring, a C3-C30 heteroaryl ring, and a C3-C30 aliphatic heterocycle, and the "ring" herein includes a monocyclic ring and a polycyclic ring; R ¹ is a substituent of the A ¹ ring, m=0-3; R ¹ is independently selected from halogen, C3-C5 cycloalkyl, C1-C3 haloalkyl, -CN, C1-C3 alkyl optionally substituted by 1-3 R ^b , C2-C6 alkenyl, C2-C6 alkynyl; R ^b is independently selected from substituted or unsubstituted C1-C3 alkoxy, where "substituted" means optionally substituted by 1-3 hydroxyl, halogen or C ₁ -C ₃ alkoxy substituents; ^R2 is a substituent of the ^A2 ring, n=0-3; ^R2 is selected from -OH, halogen, _C1 - _C3 alkoxy, _C1 - _C3 haloalkyl, -C(O) ^NHRc , -NHC(O) _CH2NHC (O) ^Rc , 3-5 membered cycloalkyl optionally substituted by 1-3 "-CN", 5-7 membered aryl optionally substituted by 1-3 "-CN"; ^Rc is _C1 - _C3 alkyl; L is a chemical bond or a divalent group selected from one or more combinations of C1-6 alkylene, saturated or partially unsaturated C3-10 cycloalkylene, -O-, -NR ^a -, and -NR ^a -C1-6 alkylene; ^Ra is independently selected from H, C1-C10 alkyl, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated 3-10 membered heterocyclic group or C6-10 aryl, CH ₂ in ^Ra can be replaced by -O- or -S-, and H in ^Ra can be replaced by hydroxyl, halogen or C1-C3 alkoxy. The sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof according to claim 1, wherein the ^A1 ring or the ^A2 ring is one of the following cyclic structures:
* indicates the site of connection with the parent core. ^R1 and ^R2 as substituents can replace any site that is chemically substitutable. ^R1 or ^R2 is selected from H, F, Cl, Br, I, hydroxyl, methyl, methoxy, trifluoromethyl, ethyl, ethynyl, cyclopropyl, cyclopentyl, cyclohexyl, cyano, formamido, cyano-substituted cyclopropyl, cyano-substituted cyclopentyl, cyano-substituted cyclohexyl, phenyl, cyano-substituted phenyl, naphthyl, cyano-substituted naphthyl and one of the following groups:
* indicates the site of attachment to the parent nucleus. The sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof according to claim 1, wherein the combination of the ^A1 ring and its substituent ^R1 is one selected from the following groups:

The combination of the ^A2 ring and its substituent ^R2 is one selected from the following groups:
* indicates the site of attachment to the parent nucleus. The sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof according to claim 1, wherein the sulfonamide compound represented by formula (I) is the following compound:

A pharmaceutical composition, comprising a sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof as described in any one of claims 1 to 4, and a pharmaceutically acceptable carrier and/or excipient, wherein the pharmaceutical composition is a solid preparation, a semisolid preparation, a liquid preparation or a gaseous preparation; the dosage form of the pharmaceutical composition is preferably an oral dosage form or an injection, wherein the oral dosage form includes capsules, tablets, pills, powders and granules, and the liquid dosage form for oral administration includes a pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture, and the injection contains a physiologically acceptable sterile aqueous or anhydrous solution, dispersion, suspension or emulsion. Use of the sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof according to any one of claims 1 to 4, or the pharmaceutical composition according to claim 5 in the preparation of an inhibitor of the interaction between WDR5 and Myc, preferably, by specifically binding to the WBM site of WDR5 to block the interaction between WDR5 and Myc. Use of the sulfonamide compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, or the pharmaceutical composition according to claim 5 in the preparation of a drug for treating or alleviating cancer, Preferably, the cancer is selected from neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, ganglioblastoma, sympathetic neuroblastoma, neurilemmoma, neurofibroma, prostate cancer, triple-negative breast cancer, nasopharyngeal cancer, esophageal cancer, laryngeal cancer, lung cancer, gastric cancer, liver cancer, colorectal cancer, cervical cancer, pancreatic cancer, bladder cancer, retinoblastoma, osteosarcoma, chondrosarcoma, chordoma, rhabdomyosarcoma, multiple myeloma, lymphoma, acute Lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia; further preferably, the cancer is selected from neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, ganglioblastoma, sympathetic neuroblastoma, neurotheca, neurotheca, neurofibroma, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, liver cancer, pancreatic cancer. A method for treating and/or preventing cancer, comprising administering to a human or animal body a therapeutically effective amount of a sulfonamide compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, and a pharmaceutical composition according to claim 5.