CN119462555A

CN119462555A - Sulfonamide compounds and uses thereof

Info

Publication number: CN119462555A
Application number: CN202411072614.0A
Authority: CN
Inventors: 白芳; 梅良和; 李凯; 曹宇; 张斌; 韩祺蕾; 孟晓冬; 王林; 任鹏璇; 王作鹏; 张向磊
Original assignee: Suzhou Zhongke New Drug Basket Biomedical Technology Co ltd; ShanghaiTech University; Childrens Hospital of Fudan University
Current assignee: Suzhou Zhongke New Drug Basket Biomedical Technology Co ltd; ShanghaiTech University; Childrens Hospital of Fudan University
Priority date: 2023-08-09
Filing date: 2024-08-06
Publication date: 2025-02-18

Abstract

The present disclosure relates to a sulfonamide compound represented by formula (I) or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, a preparation method and use thereof, and a pharmaceutical composition thereof. The present disclosure also relates to its use as a WDR5 inhibitor, which has a significant WDR5 and Myc interaction blocking effect, and is helpful for cancer treatment. ^A1 and ^A2 are ring structures, and ^R1 and ^R2 are substituents.

Description

Sulfonamide compound and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, relates to the technical field of sulfonamide derivatives, and in particular relates to a sulfonamide compound serving as an anti-neuroblastoma agent, a preparation method and application thereof.

Background

WD40 repeat domain protein 5 (WDR 5) is known to belong to the WD40 protein family, a protein containing the annular esculentus β -propeller domain. WDR5 is highly conserved among different vertebrates, with about 90% sequence identity. WDR5 was first found to be involved in regulating osteoblast differentiation during skeletal development, promoting skeletal formation. In recent years, more and more researches reveal the biological functions of WDR5, and find that the WDR5 not only regulates biological behaviors such as cell proliferation, division, apoptosis, signal transduction, gene transcription, DNA damage repair and the like under physiological states, but also is highly related to the occurrence and development of various tumors, thereby becoming a popular choice of tumor treatment targets.

The most important function of WDR5 in vivo is epigenetic regulation, which regulates histone methylation levels as a member of the Histone Methyltransferase (HMT) complex, activating transcription of target genes. WDR5 dysfunction can up-regulate the expression of various oncolytic factors, activate tumor-associated signaling pathways, and in turn enhance tumor cell proliferation, metastasis and drug resistance, driving the occurrence of epithelial-mesenchymal transition (EMT). WDR5 is also a cofactor for the myeloma viral oncogene homolog (Myc). Myc acts as a transcription factor, regulating transcription of downstream genes by binding to the promoter region of the target gene, and the limiting factor for this function is Myc's ability to localize to chromatin. It was found that about 80% of Myc binding to its downstream gene sequence of interest requires WDR5 involvement. After disruption of binding of WDR5 and Myc, myc's ability to induce pluripotent stem cells is greatly diminished, so WDR5 is a key factor in Myc-driven tumorigenesis. This mechanism was demonstrated in a variety of Myc-related tumors. In glioblastomas and neuroblastomas, WDR5 promotes Myc binding to the coactivator-associated arginine methyltransferase 1 (CARM 1) promoter, enhancing tumor cell proliferation and self-renewal capacity. In neuroblastoma, WDR5 and N-Myc co-localize to the double-microbody homologous gene 2 (MDM 2) promoter region, activate transcription of MDM2, and inhibit expression of oncostatin p 53. In cholangiocarcinoma, WDR5 maintains high levels of HIF-1α in vivo through epigenetic regulation, and simultaneously increases expression of HIF-1α through Myc in the transcriptional link, promoting EMT, metastasis and invasion of tumor cells.

There are two interaction sites on the WDR5 surface, the WDR5 binding domain (WBM) site and WDR5 interaction domain (WIN), respectively. The WBM site is a shallow and relatively hydrophobic binding pocket consisting essentially of Asn225, tyr228, leu240, phe266, val268, and Gln289, among others. Retinoblastoma binding protein 5 (RBBP 5), C-myc, L-myc, N-myc, KAT8 regulate NSL complex subunit 2 (KANSL 2), etc. all interact with WDR5 through this site. The WIN site is an arginine binding pocket and consists of amino acids such as Ala65, ser91, asp107, phe133, tyr191, tyr260, phe263 and the like. Reported interactions with WDR5 through this site have been reported for example for mixed lineage leukemia 1-4 (MLL 1-4), KAT8 regulatory NSL complex subunit 1 (KANSL), H3, cytidylylguanosine monophosphate binding protein (MBD 3), kinesin heavy chain member 2A (KIF 2A), and the like. The key biological functions of WDR5 depend on the mediation of the above two sites, such as direct binding to RBBP5, SET1/MLL, indirect binding to ASH2 (deficiency, small, or homologous) like (Drosophila) protein (ASH 2L) and DPY-30 homolog protein (DPY 30) to form a complex with histone methyltransferase catalytic activity, and through epigenetic regulation of transcription of target genes (non-patent document 4), and interaction with transcription factor Myc, through recruitment of Myc to chromatin, the binding to the promoter region of the target genes is enhanced greatly (non-patent documents 5, 6). Therefore, the design of specific protein-protein interaction blockers targeting WBM or WIN sites on WDR5 surface is a major strategy for development of novel inhibitors of WDR 5.

At present, a protein-protein interaction blocker designed and developed for the WIN site of WDR5 has strong affinity and good drug formation, but the application of the protein-protein interaction blocker is limited to the treatment of MLL gene translocation rearrangement leukemia, the effect on solid tumors is poor, and the research on the WBM site blocker is less. The Stephen research team at university of van der waals medical school, through multiple rounds of optimization modification, obtained compound No. 12 can specifically bind to WBM site, and the K _D value with WDR5 is 0.10±0.01 μm (non-patent documents 7, 8), but the antitumor effect thereof was not evaluated.

Therefore, the design and synthesis of the high-activity compound which specifically blocks the interaction of WDR5 and Myc is of great research significance and application prospect in the treatment of malignant tumor diseases, especially neuroblastoma and other diseases, and is a potential drug target and an important direction for searching lead compounds.

Non-patent reference

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 andTherapeutic Targets for Immunotherapy.Journal of immunology research 2018;2018:7394268.

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

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

5Thomas LR,Wang Q,Grieb BC,Phan J,Foshage AM,Sun Q,et al.Interaction with WDR5 promotestarget 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-MycTranscriptional 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 X.Targeting WD Repeat-Containing Protein 5(WDR5):AMedicinal Chemistry Perspective.Journal of medicinal chemistry 2021;64:10537-56.

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

Disclosure of Invention

Based on the above background, the present disclosure is directed to providing a sulfonamide compound or a pharmaceutically acceptable salt thereof, a tautomer thereof, or a stereoisomer thereof, to screen out a compound serving as a WDR5 inhibitor having excellent performance in terms of performance such as effectiveness, safety, and selectivity.

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

Wherein:

The A ¹ ring and the A ² ring are each independently selected from one of a C6-C30 aryl ring, a C6-C30 aliphatic hydrocarbon ring, a C3-C30 heteroaryl ring, a C3-C30 aliphatic heterocyclic ring, where "ring" includes monocyclic and polycyclic rings;

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 with 1-3R ^b, C2-C6 alkenyl, C2-C6 alkynyl, R ^b is independently selected from substituted or unsubstituted C1-C3 alkoxy, where "substituted" means optionally substituted with 1-3 hydroxy, halogen or a substituent of C ₁-C₃ alkoxy;

R ² is a substituent of the A ² ring, n=0-3;R ² is selected from-OH, halogen, C ₁-C₃ alkoxy, C ₁-C₃ haloalkyl, -C (O) NHR ^c、-NHC(O)CH₂NHC(O)R^c, 3-5 membered cycloalkyl optionally substituted by 1-3 "-CN", 5-7 membered aryl optionally substituted by 1-3 "-CN";

L is a bond or a divalent group formed by combining one or more selected from C1-6 alkylene, saturated or partially unsaturated C3-10 cycloalkylene, -O-, -NR ^a-、-NR^a -C1-6 alkylene, R ^a is independently selected from H, C-C10 alkyl, saturated or partially unsaturated C3-6 cycloalkyl, saturated or partially unsaturated 3-10 heterocyclyl or C6-10 aryl, CH ₂ in R ^a may be replaced by-O-or-S-, and H in R ^a may be replaced by hydroxy, halogen or C1-C3 alkoxy.

The compound disclosed by the disclosure is a compound specifically bound to the WBM locus of WDR5, and can block the interaction between WDR5 and Myc, so that Myc-mediated oncogene transcription is regulated and controlled, and the tumor treatment effect is achieved. Pharmacological experiments show that the compound disclosed by the embodiment of the invention has stronger affinity to WDR5 protein, can obviously inhibit proliferation of cells in a MYCN high-amplified neuroblastoma cell line, has no larger influence on survival of human embryonic kidney cells HEK293T, and shows good selectivity and safety. According to the analysis result of the crystal complex structure, the compound disclosed by the disclosure can specifically bind to the WBM domain of WDR5 to block the interaction between WDR5 and Myc, thereby exerting anti-tumor activity.

In an alternative embodiment of the present disclosure, the A ¹ 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, the A ² 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, and at least one halogen is present as a substituent in R ¹ and R ².

In an alternative embodiment of the present disclosure, the a ¹ ring or a ² ring is one of the ring structures selected from the group consisting of:

* Represents a site of attachment to the parent nucleus, and R ¹ and R ² as substituents may be substituted at any site that is chemically substitutable.

In an alternative embodiment of the disclosure, R ¹ or R ² is one selected from H, F, cl, br, I, hydroxy, methyl, methoxy, trifluoromethyl, ethyl, ethynyl, cyclopropyl, cyclopentyl, cyclohexyl, cyano, carboxamido, cyano-substituted cyclopropyl, cyano-substituted cyclopentyl, cyano-substituted cyclohexyl, phenyl, cyano-substituted phenyl, naphthyl, cyano-substituted naphthyl, and:

* Represents the site of ligation to the parent nucleus.

In an alternative embodiment of the present disclosure, the combination of the a ¹ ring and its substituent R ¹ is one selected from the group consisting of:

In an alternative embodiment of the present disclosure, the combination of the a ² ring and its substituent R ² is one selected from the group consisting of:

* Represents the site of ligation to the parent nucleus.

The compound disclosed by the invention can be used as a WDR5-Myc interaction blocker, and in particular, pharmacological experiment results show that the compound disclosed by the invention can specifically bind to a WBM domain of WDR5 to block the interaction between WDR5 and Myc. Based on the application, the compound disclosed by the disclosure can be applied to the preparation of medicines for treating diseases related to abnormal activation of Myc, RBBP5, KANSL and other related signal paths caused by abnormal expression of WDR 5.

It is known that diseases associated with abnormal activation of signal pathways associated with WDR5 such as Myc, RBBP5, KANSL and the like include, but are not limited to, neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroblastoma, sympathogenic neuroblastoma, schwannoma, neuroblastoma, 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, osteogenic sarcoma, chondrosarcoma, chordoma, rhabdomyosarcoma, multiple myeloma, lymphoma, acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphoblastic leukemia and chronic myelogenous leukemia, and the like, and are particularly suitable for treatment and alleviation of neurogenic tumors including neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroblastoma, neuroblastoma, sympathogenic tumor, neurofibroma.

As typical compounds of the present disclosure, the following compounds may be mentioned, but are not limited thereto:

Another aspect of the present disclosure provides a pharmaceutical composition, which is characterized in that the pharmaceutical composition comprises the sulfonamide compound of the present disclosure 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, wherein the oral dosage form comprises capsules, tablets, pills, powder and granules. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures, and the injection comprises 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 undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio.

In the present disclosure, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or excipient for delivering the active substances of the present disclosure, or physiologically acceptable salts thereof, to an animal or human. The carrier may be a liquid or a solid.

In the present disclosure, the pharmaceutical composition comprises a safe and effective amount (e.g., 0.001-99.9 parts by weight, may be 0.01-99 parts by weight, and may be 0.1-90 parts by 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 parts by weight.

Or the pharmaceutical composition of the present disclosure contains 0.001 to 99.9wt%, more preferably 0.01 to 99 wt%, and still more preferably 0.1 to 90 wt% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, based on the total weight, and a pharmaceutically acceptable carrier or excipient, wherein the total weight of the composition is 100 wt%.

In another alternative embodiment, the compound of formula (I) is present in an optional proportion with a pharmaceutically acceptable carrier, excipient or slow release agent, wherein formula (I) is present as active ingredient in an amount of more than 70% by weight, the remainder being present in an amount of 0.5 to 30% by weight, or more preferably 1 to 15% by weight, or most preferably 2 to 10% by weight.

The pharmaceutical compositions of the present disclosure may be in various dosage forms comprising a unit dose of 0.5mg to 200mg, may be 2mg to 100mg, and more may be 5mg to 50mg of the compound of formula (I), enantiomer, racemate, pharmaceutically acceptable salt or mixture thereof.

When the pharmaceutical composition contains an additional pharmaceutically active ingredient for treating or preventing cancer, the amount of the active ingredient may be generally the conventional amount or less in the art.

The pharmaceutical compositions of the present disclosure may be in a variety of forms, such as tablets, capsules, powders, syrups, solutions, suspensions, and aerosols, etc., wherein the compounds of formula (I) may be presented in a suitable solid or liquid carrier or diluent. The pharmaceutical compositions of the present disclosure may also be stored in a suitable injectable or instillation disinfection device. The pharmaceutical composition may also contain odorants, flavoring agents, etc.

The compounds of formula (I) or pharmaceutical compositions comprising the compounds of formula (I) of the present disclosure may be administered to mammals, including humans, clinically via oral, nasal, dermal, pulmonary or gastrointestinal routes of administration. An alternative route of administration is oral. The optional daily dose is 0.5mg-100mg/kg body weight, and can be administered once or in divided doses. Regardless of the method of administration, the optimal dosage for an individual will depend on the particular treatment. Typically starting from a small dose, the dose is gradually increased until the most appropriate 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 condition to be treated. However, generally, satisfactory results are obtained when the compounds of the present disclosure are administered at a dose of about 1-100mg/kg animal body weight per day, preferably at 1-3 divided doses per day, or in a sustained release form. For most large mammals, the total daily dose is about 5-500mg, preferably about 10-250mg. Dosage forms suitable for oral administration comprise about 1-100mg of the active compound intimately mixed with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen can be adjusted to provide the optimal therapeutic response. For example, separate doses may be administered several times per day, or the dose may be proportionally reduced, as dictated by the urgent need for the treatment of the condition.

The compounds or pharmaceutically acceptable salts thereof and compositions thereof may be administered orally as well as intravenously, intramuscularly or subcutaneously. From the standpoint of ease of preparation and administration, alternative pharmaceutical compositions are solid state compositions, especially tablets and solid filled or liquid filled capsules. 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 glycols, nonionic surfactants and edible oils (e.g., corn, peanut and sesame oils) as are appropriate to the nature of the active ingredient and the particular mode of administration desired. Adjuvants commonly used in the preparation of pharmaceutical compositions may also be advantageously included, for example, flavourings, colours, preservatives and antioxidants such as vitamin E, vitamin C, BHT and BHA.

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

Pharmaceutical forms suitable for injection include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, these forms must be sterile and must be fluid to facilitate the discharge of the fluid from the syringe. Must be stable under the conditions of manufacture and storage and must be able to prevent the contaminating effects of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, alcohols (such as glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

The compounds of formula (I) or pharmaceutically acceptable salts and compositions thereof may also be administered in combination with other active ingredients or drugs for the treatment or prophylaxis of cancer diseases. When two or more drugs are administered in combination, there is generally an effect superior to the effect of the two drugs administered separately.

In another aspect of the present disclosure, there is provided the use of a compound of the present disclosure and a corresponding pharmaceutical composition in the manufacture of a medicament for the treatment or alleviation of cancer. The compositions and methods provided by the present disclosure are useful for treating a variety of cancers, including prostate, breast, brain, skin, cervical cancer, testicular cancer, and the like. More specifically, cancers treatable by the compositions and methods of the present disclosure include, but are not limited to, tumor types such as astrocyte, breast, cervical, colorectal, endometrial, esophageal, stomach, head-neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid cancers and sarcomas. More specifically, these compounds are useful in the treatment of sarcomas (hemangiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma in the heart, bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondria, mesothelioma, gastrointestinal tract (esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagon, gastrinoma, carcinoid, VIPoma), gastrointestinal tract, Small intestine (adenocarcinoma, lymphoma, carcinoid, kaposi's sarcoma, smooth myoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, chorionic adenoma, hamartoma, smooth myoma), genitourinary tract, kidney (adenocarcinoma, nephroblastoma (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryo carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, adenoma-like tumor, lipoma), liver (hepatocellular carcinoma), kidney (hepatocellular carcinoma), Bile duct cancer, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma, biliary tract, gall bladder cancer, ampulla cancer, bile duct cancer, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, ewing's sarcoma, malignant lymphoma (reticulocytosoma), multiple myeloma, malignant giant cell tumor, bone chronicity tumor (osteochondral sarcoma), benign tumor, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumor, nervous system, skull (osteoma, hemangioma, granuloma, xanthoma, osteomyelitis), meningioma (meningioma, glioma), brain (astrocytoma ), Medulloblastoma, glioma, ependymoma, germ cell tumor (pineal tumor), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), spinal fibroma, meningioma, glioma, sarcoma, gynaecology, uterus (endometrial carcinoma), cervix (cervical carcinoma, precancerous cervical dysplasia), ovary (ovarian carcinoma (serous cystic adenocarcinoma, mucinous cystic adenocarcinoma, unclassified carcinoma), granulosa sheath 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 (carcinoma), hematology: blood (acute and chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic 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, cutaneous fibroma, lupus erythematosus, psoriasis, and adrenal gland: neuroblastoma. In certain embodiments, the cancer is diffuse large B-cell lymphoma (DLBCL).

In an alternative embodiment, there is provided the use of a compound having a structure represented by the following formula (I) or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof as a medicament for treating neurogenic tumors, and the use of a compound having a structure represented by the following formula (I) or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof as a medicament for treating and alleviating neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroma, ganglioneuroblastoma, sympathogenic neuroblastoma, schwannoma, neurosheath tumor, neurofibroma.

In alternative embodiments, the cancer is selected from the group consisting of neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroblastoma, sympathogenic neuroblastoma, schwannoma, 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, osteogenic sarcoma, chondrosarcoma, chordoma, rhabdomyosarcoma, multiple myeloma, lymphoma, acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and in alternative embodiments the cancer is selected from the group consisting of neurogenic tumors including neuroblastoma, gangliocytoma, ganglioblastoma, ganglioneuroblastoma, sympathogenic neuroblastoma, schwannoma, neurofibroma.

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

Interpretation of the terms

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

Definition of the definition

Unless defined otherwise hereinafter, all technical and scientific terms used herein are intended to be identical to what is commonly understood by one of ordinary skill in the art. References to techniques used herein are intended to refer to techniques commonly understood in the art, including variations of those that are obvious to those skilled in the art or alternatives to equivalent techniques. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present disclosure.

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

As used herein, the term "alkylene" means a saturated divalent hydrocarbon group, which may represent 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 means a cycloalkyl group, a cycloalkenyl group, a cycloalkynyl group, etc. having a cyclic structure composed of C and H, wherein C is substituted with a hetero atom and is an aliphatic heterocyclic group, and also includes bridged and spiro forms described later.

As used herein, the term "alkyl" is defined as a linear or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group 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) optionally substituted with 1 or more (such as 1 to 3) suitable substituents such as halogen (in which case the group is referred to as "haloalkyl") (e.g., CH₂F、CHF₂、CF ₃、CCl₃、C₂F₅、C₂Cl₅、CH₂CF₃、CH₂Cl or-CH ₂CH₂CF₃, etc.). The term "C1-4 alkyl" refers to a linear or branched aliphatic hydrocarbon chain of 1 to 4 carbon atoms (i.e., 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 _2-6 alkenyl"). The alkenyl group is, for example, vinyl, 1-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 a compound of the present disclosure contains an alkenyl group, the compound may exist in pure E (ipsilateral (entgegen)) form, pure Z (ipsilateral (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 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 multicyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, 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 as1 to 3) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term "C _3-6 cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring of 3 to 6 ring-forming carbon atoms (e.g., cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), optionally substituted with 1 or more (such as1 to 3) suitable substituents, for example methyl substituted cyclopropyl.

As used herein, the terms "cycloalkylene", "cyclic hydrocarbon" and "hydrocarbon ring" refer to a saturated (i.e., "cycloalkylene" and "cycloalkyl") or unsaturated (i.e., having one or more double and/or triple bonds within the ring) mono-or polycyclic hydrocarbon ring having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring carbon atoms, including, but not limited to, cyclopropyl (cyclo), (cyclobutyl (cyclo) ene, (cyclopentyl (cyclo) ene), (cyclohexyl (cyclo) ene), (cycloheptyl (cyclo), (cyclooctyl (cyclo), (cyclonon) (cyclohexenyl (cyclo) ene), 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 and/or triple bonds within the ring) cyclic group having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring atoms, at least one of which is a heteroatom selected from N, O and S, and the remaining ring atoms being C. For example, a "3-10 membered (sub) heterocyclic (group)" is a saturated or partially unsaturated (sub) heterocyclic (group) 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 heterocyclic (yl) groups include, but are not limited to, ethylene oxide, (methylene) aziridinyl, (methylene) azetidinyl (azetidinyl), (methylene) oxetanyl (oxetanyl), (methylene) tetrahydrofuranyl, (methylene) dioxolyl (dioxolinyl), (methylene) pyrrolidinyl, (methylene) pyrrolidonyl, (methylene) imidazolidinyl, (methylene) pyrazolidinyl, (methylene) pyrrolinyl, (methylene) tetrahydropyranyl, (methylene) piperidinyl, (methylene) morpholinyl, (methylene) dithianyl (dithianyl), (methylene) thiomorpholinyl, (methylene) piperazinyl or (methylene) trithianyl (trithianyl). The groups also encompass 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, and the like). The heterocyclylene and heterocyclic (groups) may be optionally substituted with one or more (e.g., 1,2, 3 or 4) suitable substituents.

As used herein, the terms "(arylene) and" aromatic ring "refer to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated pi-electron system. For example, as used herein, the terms "C _6-10 (arylene) and" C _6-10 aromatic ring "mean an aromatic group containing 6 to 10 carbon atoms, such as a phenyl (phenylene) or a naphthyl (phenylene) group. The aryl (ene) and aromatic rings 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 "(arylene) heteroaryl" and "heteroaryl ring" refer to a monocyclic, bicyclic or tricyclic aromatic ring system having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, particularly 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 the same or different, such as oxygen, nitrogen or sulfur), and which may additionally be benzo-fused in each case. In particular, "(arylene) heteroaryl" or "heteroaryl ring" is selected from thienyl (ene) furyl (ene) pyrrolyl (ene) oxazolyl (ene) thiazolyl (ene) imidazolyl (ene) pyrazolyl (ene) isoxazolyl (ene) isothiazolyl (ene) oxadiazolyl (ene) triazolyl (ene) thiadiazolyl and the like, and their benzo derivatives, or pyridyl (ene) pyridazinyl (ene) pyrimidinyl (ene) pyrazinyl (ene) triazinyl and the like, and their benzo derivatives.

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

As more specific terms are explained as follows:

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

"Alkenyl" refers to a straight or branched chain monovalent hydrocarbon radical of 2 to 12 carbon atoms, or 2 to 8 carbon atoms, or 2 to 6 carbon atoms, or 2 to 4 carbon atoms, wherein at least one C-C is an sp ² double bond, wherein the alkenyl group may be independently optionally substituted with 1 or more substituents described in this disclosure, specific examples of which include, but are not limited to, vinyl, allyl, and the like. Alkenyl groups may be optionally substituted or unsubstituted.

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

"Spirocycloalkyl" refers to a 5 to 18 membered, two or more cyclic structure, and monocyclic polycyclic groups sharing one carbon atom (called spiro atom) with each other, containing 1 or more double bonds within the ring, but no ring has a completely conjugated pi-electron aromatic system. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The spirocycloalkyl group is classified into a single spiro group, a double spiro group or a multiple spirocycloalkyl group according to the number of common spiro atoms between rings, preferably single spiro group and double spirocycloalkyl group, preferably 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered. Non-limiting examples of "spirocycloalkyl" include, but are not limited to:

"fused ring alkyl" refers to an all-carbon polycyclic group containing two or more cyclic structures sharing a pair of carbon atoms with each other, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicycloalkyl group. Non-limiting examples of "fused ring alkyl" include, but are not limited to:

"bridged cycloalkyl" means an aromatic system having 5 to 18 members, containing two or more cyclic structures, sharing two all-carbon polycyclic groups with one another that are not directly attached to a carbon atom, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi electron, preferably 6 to 12 members, more preferably 7 to 10 members. Cycloalkyl groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are 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 cycloalkyl, non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, and the like.

"Heterocyclyl", "heterocycle" or "heterocyclic" are used interchangeably herein and refer to a saturated or partially unsaturated, monocyclic, bicyclic or tricyclic, non-aromatic heterocyclic group containing 3 to 12 ring atoms, at least one of which is a heteroatom such as oxygen, nitrogen, sulfur, and the like. Preferably having a5 to 7 membered mono-or 7 to 10 membered bi-or tri-ring, which may contain 1,2 or 3 atoms selected from nitrogen, oxygen and/or sulphur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydropyranyl, 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 heterocyclyl. The heterocyclyl group may be optionally substituted or unsubstituted.

"Spiroheterocyclyl" refers to a 5 to 18 membered, two or more cyclic structure, polycyclic groups sharing one atom between each other between single rings, containing 1 or more double bonds within the ring, but no ring having a fully conjugated pi-electron aromatic system wherein one or more ring atoms are selected from nitrogen, oxygen, sulfur or a heteroatom of S (O) _m, the remaining ring atoms being carbon, m=1 or 2. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The spiroheterocyclyl groups are classified into a single spiroheterocyclyl group, a double spiroheterocyclyl group or a multiple spiroheterocyclyl group according to the number of common spiro atoms between rings, and preferably a single spiroheterocyclyl group and a double spiroheterocyclyl group. More preferably a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered single spiro heterocyclic group. Non-limiting examples of "spiroheterocyclyl" include, but are not limited to:

"fused heterocyclyl" refers to an all-carbon polycyclic group containing two or more cyclic structures sharing a pair of atoms with each other, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron aromatic system in which one or more of the ring atoms is selected from nitrogen, oxygen, sulfur or a heteroatom of S (O) _m, the remaining ring atoms being carbon, m=1 or 2. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The number of constituent rings may be classified as 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 heterocyclyl" include, but are not limited to:

"bridged heterocyclyl" means a 5-to 18-membered, polycyclic group containing two or more cyclic structures sharing two atoms not directly attached to each other, one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron aromatic system in which one or more of the ring atoms is selected from nitrogen, oxygen, sulfur or a heteroatom of S (O) _m, the remaining ring atoms being carbon, m=1 or 2. Preferably 6 to 14 membered, more preferably 7 to 10 membered. Heterocyclic groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclyl" include, but are not limited to:

"aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be linked together in a fused manner. The term "aryl" includes aromatic groups such as phenyl, naphthyl, tetrahydronaphthyl. Preferably aryl is C ₆-C₁₀ aryl, more preferably aryl is phenyl and naphthyl, most preferably phenyl. Aryl groups may be substituted or unsubstituted. The "aryl" may be fused to a heteroaryl, heterocyclyl, or cycloalkyl group, wherein the aryl ring is attached to the parent structure, 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. Examples of "heteroaryl" include, but are 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, quinolinyl, indazolyl, benzisothiazolyl, benzoxazolyl and benzisoxazolyl. Heteroaryl groups may be optionally substituted or unsubstituted. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached 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. Alkoxy of C ₁-C₆ is a preferred choice. Examples 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 halo 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-dichloroethyl, 1, 2-dichloropropyl, and the like.

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

As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or unsaturated mono-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 further comprise 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 attached to the remainder of the molecule through the nitrogen atom in the nitrogen-containing heterocycle and any remaining ring atoms, which is optionally benzo-fused, and which may be attached to the remainder of the molecule through the nitrogen atom in the nitrogen-containing heterocycle and any carbon atom in the fused benzene ring.

The term "substitution" means that one or more (e.g., one, two, three, or four) hydrogens on the designated atom are replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution forms a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in 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 list of substituents, one or more hydrogens on the carbon (to the extent any hydrogens are present) may be replaced with an independently selected optional substituent, alone and/or together. If the nitrogen of a substituent is described as optionally substituted with one or more of the list of substituents, 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 "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.

The term "one or more" as used herein means 1 or more than 1, such as 2, 3,4, 5 or 10, under reasonable conditions.

As used herein, unless indicated, the point of attachment of a substituent may be from any suitable position of the substituent.

When the bond of a substituent is shown as a bond through the ring connecting two atoms, then such substituent may be bonded to any ring-forming 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 prevailing 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 (² H), Tritium (³ H)), isotopes of carbon (e.g., ¹¹C、¹³ C and ¹⁴ C), isotopes of chlorine (e.g., ³⁶ Cl), isotopes of fluorine (e.g., ¹⁸ F), isotopes of iodine (e.g., ¹²³ I and ¹²⁵ I), isotopes of nitrogen (e.g., ¹³ N and ¹⁵ N), isotopes of oxygen (e.g., ¹⁵O、¹⁷ O and ¹⁸ O), isotopes of phosphorus (e.g., ³² P), and isotopes of sulfur (e.g., ³⁵ S). Certain isotopically-labeled compounds of the present disclosure (e.g., those into which a radioisotope is incorporated) are useful in pharmaceutical and/or substrate tissue distribution studies (e.g., assays). The radioisotope tritium (i.e., ³ H) and carbon-14 (i.e., 14C) are particularly useful for this purpose because of their ease of incorporation and ease of detection. Substitution with positron emitting isotopes (such as ¹¹C、¹⁸F、¹⁵ O and ¹³ N) can be used in Positron Emission Tomography (PET) studies to examine substrate receptor occupancy. isotopically-labeled compounds of the present disclosure can be prepared by processes analogous to those described in the accompanying schemes and/or in the examples and preparations by substituting an appropriate isotopically-labeled reagent for the previously employed non-labeled reagent. Pharmaceutically acceptable solvates of the present disclosure include those in which the crystallization solvent may be isotopically substituted, e.g., 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 to 3 hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable when bound to carbon atoms having unsaturated (e.g., olefinic) bonds.

"Substituted" or "substituted" as used herein, unless otherwise indicated, means that the group may be substituted with one or more groups selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate 、＝O、-C(O)R^b、-OC(O)R^b、-NR^bR^b、-C(O)NR^bR^b、-NR^bC(O)R^b、-S(O)NR^bR^b, or-S (O) ₂NR^bR^b, where R ^b is as defined in formula (I).

As used herein, a "therapeutically effective dose" of a compound refers to an amount sufficient to ameliorate or somehow reduce symptoms, stop or reverse progression of a condition. Such doses may be administered as a single dose or may be administered according to a regimen so as to be effective. As used herein, "treating" refers to ameliorating or otherwise altering the condition, disorder, or symptom or pathology of a disease in a patient in any manner.

As used herein, "ameliorating a symptom of a particular disease by use of a particular compound or pharmaceutical composition" refers to any reduction, whether permanent or temporary, persistent or temporary, attributable to or associated with the use of the composition.

The compounds of the present disclosure may contain asymmetric centers or chiral centers and thus different stereoisomers are present. All stereoisomers of the compounds of the present disclosure, including, but in no way limited to, diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, form part of the present disclosure. The terms "racemic mixture" and "racemate" refer to a mixture of two enantiomers in equimolar amounts, lacking optical activity.

"Tautomer" or "tautomeric form" refers to isomers of structures of different energies that can be interconverted by a low energy barrier. Individual stereochemical isomers of the compounds of the present disclosure, or enantiomers, diastereomers, or mixtures of geometric isomers thereof, are all within the scope of the present disclosure.

Solid lines may be used hereinSolid wedge shapeOr virtual wedge shapeDepicting the chemical bond of the compounds of the present disclosure. The use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom (e.g., particular enantiomers, racemic mixtures, etc.) are included. The use of a solid or virtual wedge to depict a bond to an asymmetric carbon atom is intended to indicate the presence of the stereoisomers shown. When present in a racemic mixture, real and imaginary wedges are used to define the relative stereochemistry, not the absolute stereochemistry. Unless otherwise indicated, compounds of the present disclosure are intended to exist as stereoisomers (which include cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotamers, 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 diastereomeric pairs).

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

It will also be appreciated that certain compounds of the present disclosure may exist in free form for use in therapy, 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, clathrates, or prodrugs that, upon administration to a patient in need thereof, are capable of providing the compounds of the present disclosure, or metabolites or residues thereof, directly or indirectly. Thus, when reference is made herein to "a compound of the present disclosure," it is also intended to encompass the various derivative forms of the compounds described above.

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 coordination bonds.

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

Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, benzathine 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 the compounds of the present disclosure are known to those skilled in the art.

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

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

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides, as nitrogen requires available lone pairs to oxidize to oxides, and those skilled in the art will recognize nitrogen-containing heterocycles 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 and include oxidizing heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes (dioxirane) such as dimethyl dioxirane.

Also included within the scope of the present disclosure are metabolites of the compounds of the present disclosure, i.e., substances that form in vivo upon administration of the compounds of the present disclosure. Such products may result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the compound being administered. Accordingly, the present disclosure includes metabolites of the compounds of the present disclosure, including compounds made by a method of contacting a compound of the present disclosure with a mammal for a time sufficient to produce a metabolite thereof.

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

The present disclosure also encompasses compounds of the present disclosure that contain a protecting group. During any process of 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 of interest, 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. The protecting group may be removed at a suitable subsequent stage using methods known in the art.

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

The WDR5/Myc interaction blocker with a novel structure is provided, and test results show that the derivative has excellent anti-neuroblastoma activity, excellent safety and selectivity, and can be used for preparing medicines for treating cancers, especially neuroblastoma and other diseases.

Drawings

FIG. 1 is a schematic diagram of the complex crystal structure of a compound of the present disclosure with WDR 5.

Detailed Description

The method of the present disclosure is described below by way of specific examples to facilitate understanding and grasping of the technical solution of the present disclosure, but the present disclosure is not limited thereto. The ¹ H NMR spectra in the examples below were determined using a Bruker instrument (400 MHz) and the chemical shifts were expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. ¹ H NMR representation method: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, q=four-fold peak, m = multiple peak(s). If coupling constants are provided, they are in Hz.

The mass spectrum is measured by an LC/MS instrument, and the ionization mode is ESI. High performance liquid chromatograph model No. Agilent 1260, siemens flight U3000, chromatographic column No. Waters xbrige C18 (4.6X105 mm,3.5 μm), mobile phase A ACN, B Water (0.1% H ₃PO₄), flow rate 1.0mL/min, gradient ：5％A for 1min,increase to 20％A within 4min,increase to 80％A within 8min,80％A for2min,back to 5％A within 0.1min; wavelength 220nm, column oven 35 ℃. The thin layer chromatography silica gel plate is a smoke table yellow sea HSGF254 or Qingdao GF254 silica gel plate, the specification of the silica gel plate used by the Thin Layer Chromatography (TLC) is 0.2mm-0.3mm, and the specification of the thin layer chromatography separation and purification product is 0.4mm-0.5mm. Column chromatography generally uses tobacco stage yellow sea silica gel 200-300 mesh silica gel as carrier.

In the following examples, unless otherwise indicated, all temperatures are in degrees celsius and unless otherwise indicated, various starting materials and reagents are either commercially available or synthesized according to known methods, and are used without further purification, and unless otherwise indicated, commercially available manufacturers include, but are not limited to, the national pharmaceutical community, the carbofuran technologies, the tencel (Shanghai) chemical industry development limited, the Shanghai Pico pharmaceutical technologies limited, the Shanghai Michelson chemical technologies limited, and the like. CD ₃ OD is deuterated methanol, CDCl ₃ is deuterated chloroform, DMSO-d ₆ is deuterated dimethyl sulfoxide, pd ₂(dba)₃ is tris (dibenzylideneacetone) dipalladium, pd (dppf) Cl ₂ is [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride, xantphos is 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene, XPhos is 2-dicyclohexylphosphorus-2, 4, 6-triisopropylbiphenyl, HATU is 2- (7-benzotriazol) -N, N, N ', N ' -tetramethylurea hexafluorophosphate, EDCI is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, HOBT is 1-hydroxybenzotriazole, BINAP is 1,1' -binaphthyl-2, 2' -bisdiphenylphosphine, DCM is dichloromethane, PE is petroleum ether is EA is ethyl acetate, methanol is DMF is N, N-dimethylformamide, HPLC is a thin layer chromatography is performed at a high-to a thin layer chromatography-to the point of origin of the thin layer chromatography which is located at a distance from the TLC of 4980. The hydrogen atmosphere is a reaction flask connected to a hydrogen balloon of about 1L volume.

The examples are not particularly described, and the solution in the reaction is an aqueous solution. The temperature of the reaction was room temperature, 20 ℃ to 30 ℃, unless otherwise specified in the examples. The monitoring of the reaction progress in the examples adopts Thin Layer Chromatography (TLC), the developing agent used in the reaction, the system of column chromatography eluent used for purifying the compound or the developing agent system of the thin layer chromatography comprises A petroleum ether and ethyl acetate system, B methylene dichloride and methanol system, C normal hexane and ethyl acetate, wherein the volume ratio of the solvent is different according to the polarity of the compound, and small amount of acid or alkaline reagent such as acetic acid or triethylamine and the like can be added for adjustment. The reagent for providing alkaline conditions is selected from organic base or inorganic base, wherein the organic base is one or more of triethylamine, N-diisopropylethylamine, N-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamino, sodium tert-butoxide, sodium methoxide and potassium tert-butoxide, the inorganic base is one or more of sodium hydride, potassium phosphate, sodium carbonate, potassium acetate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium bicarbonate and lithium hydroxide, and 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, tetra-triphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1 '-bis (diphenylphosphino) ferrocene ] palladium dichloride (Pd (dppf) Cl ₂), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex, bis-triphenylphosphine palladium dichloride (Pd (PPh ₃)₂Cl₂) and tris (dibenzylideneacetone) dipalladium (Pd ₂(dba)₃);

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

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

the oxidant is one or more of potassium permanganate, manganese dioxide, potassium dichromate, sodium dichromate and potassium osmium;

The above reaction is preferably carried out in a solvent selected from one or more of N, N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, 1, 4-dioxane, water, tetrahydrofuran, methylene chloride, 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

First step2, 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℃and reacted at room temperature for 2 hours, and TLC showed disappearance of starting material. The reaction solution was directly concentrated to give the title compound IN-1 (320 mg, crude) as a white solid, which was used directly IN the next step.

Example 1

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

First step 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, the reaction was allowed to proceed for 18 hours at 140℃after the addition, the reaction mixture was naturally cooled to room temperature, phosphorus pentachloride (18.4 g,88.4 mmol) was slowly added, and there was a severe exotherm (note of blowout prevention) and the addition was allowed to proceed to 120℃after the addition, and the reaction was continued for 3 hours, TLC showed the disappearance of starting material. The reaction solution was cooled to room temperature, slowly added dropwise to ice water (100 mL), dichloromethane (20 ml×3) was added for extraction, the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 1b (9.3 g, crude) as a brown yellow oil, which was used directly in the next step.

Second step 2, 4-dimethylthiazole-5-sulfonamide 1c

Compound 1b (8.8 g, crude product) was dissolved in 1, 4-dioxane (50 mL), cooled to 0deg.C, ammonia (15 mL) was slowly added dropwise, and the reaction was continued for 10 min at 0deg.C after the addition, and TLC showed the disappearance of starting material. The reaction solution was slowly poured into ice water (100 mL), acidified to ph=3 by adding dilute hydrochloric acid (3N), extracted by adding dichloromethane (30 ml×3), the organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound 1c (7.2 g, crude) as a brown yellow oil, which was used directly in the next step.

Third step 2, 4-dichloro-N- ((2, 4-dimethylthiazol-5-yl) sulfonyl) benzamide 1

Compound 1c (230 mg, crude) was dissolved IN ethyl acetate (10 mL), 4-dimethylaminopyridine (5 mg,0.04 mmol), triethylamine (2 mL,14.4 mmol) and intermediate IN-1 (320 mg, crude) IN toluene (15 mL) were added sequentially at room temperature, and after addition, the reaction was continued at room temperature for 10 hours and TLC showed the disappearance of starting material. The reaction mixture was quenched with water (10 mL), extracted with ethyl acetate (10 mL. Times.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 give 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)

¹H NMR(400MHz,DMSO-d₆ ) Delta 7.75 (d, j=2.0 hz, 1H), 7.56-7.50 (m, 2H), 2.69 (s, 3H), 2.58 (s, 3H), (sulfonamide active hydrogen not shown)

Example 2

N ¹ - ((5-bromothiophen-2-yl) sulfonyl) -5- (1-cyanocyclopentyl) -2-hydroxy-N ³ -methyl isophthalamide 2

First step 3-bromo-5-formyl-2-hydroxybenzoic acid 2b

5-Formyl salicylic acid 2a (10.10 g,60.19 mmol) was dissolved in DMF (100 mL), cooled to 0deg.C, N-bromo-succinimide (NBS) (10.87 g,61.07 mmol) was slowly added thereto, the reaction was continued for 2 hours after slowly warming to 70deg.C, TLC monitoring (DCM/MeOH=5/1, R _f =0.2) was complete, the reaction solution was quenched with water (300 mL), extracted with ethyl acetate (100 mL x 2), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give the title compound 2b (15.02 g, crude) as a yellow solid, which was used directly in the next step.

Second step 3-bromo-5-formyl-2-methoxybenzoic acid methyl ester 2c

Compound 2b (15.02 g, crude) was added to DMF (100 mL), potassium carbonate (25.34 g,183.62 mmol) and methyl iodide (21.73 g,153.05 mmol) were added, the reaction was stirred at room temperature for 18 hours, TLC was checked (DCM/meoh=5/1, r _f =0.2) starting material was reacted completely, the reaction was quenched with water (300 mL), ethyl acetate (100 mL x 3) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude was purified by silica gel column chromatography (PE/ea=10/1) to give the title compound 2c (5.40 g, 32% in two steps) as a white solid

¹H NMR(400MHz,CDCl₃)δ9.93(s,1H),8.26(dd,J=4.8,2.4Hz,2H),4.01(s,3H),3.97(s,3H).

Third step 3-bromo-5- (hydroxymethyl) -2-methoxybenzoic acid methyl ester 2d

Compound 2C (10.00 g,36.62 mmol) was added to methanol (200 mL), cooled to 0-5 ℃, sodium borohydride (813 mg,21.9 mmol) was slowly added thereto, stirring was continued for 30 minutes after the addition, TLC detection (PE/ea=4/1, r _f =0.6) starting material reacted completely, the reaction solution was quenched with water (200 mL), extracted with ethyl acetate (100 mL x 3), and the organic phases were combined. Dried over anhydrous sodium sulfate and concentrated to give the title compound 2d (10.50 g, crude) as a yellow oil, which was used directly in the next step.

Fourth step methyl 3-bromo-2-methoxy-5- (((methylsulfonyl) oxy) methyl) benzoate 2e

Compound 2d (10.50 g, crude) was added to dichloromethane (100 mL) to which triethylamine (5.50 g,54.4 mmol) was added. The reaction was cooled to 0-5 ℃ and methylsulfonyl chloride (4.68 g,40.8 mmol) was slowly added dropwise, the reaction was stirred for 30min after addition, TLC monitoring (PE/ea=4/1, r _f =0.2) was complete, the reaction was quenched with saturated brine (100 mL), extracted with dichloromethane (50 mL x 2), the organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give the title compound 2e (11.82 g, crude) as a yellow oil, which was used directly in the next step.

Fifth step 3-bromo-5- (cyanomethyl) -2-methoxybenzoic acid methyl ester 2f

Compound 2e (11.82 g, crude product) was added to acetonitrile (100 mL), potassium carbonate (9.24 g,66.95 mmol) and trimethylcyanosilane (5.00 g,50.39 mmol) were added thereto, the reaction mixture was stirred at room temperature for 18 hours, TLC was checked (PE/EA=4/1, R _f =0.6) and was reacted completely, the reaction mixture was quenched with water (100 mL), ethyl acetate (100 mL x 3) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (PE/EA=10/1) to give the title compound 2f (4.24 g, three-step yield 40.76%)

Sixth step 2g of 3-bromo-5- (1-cyanocyclopentyl) -2-methoxybenzoic acid methyl ester

Compound 2f (104 mg,0.385 mmol) was added to dry tetrahydrofuran (5 mL), the reaction solution was cooled to 0 ℃, a solution of bis (trimethylsilyl) sodium amide in tetrahydrofuran (0.45 mL,0.90mmol,2 m/L) was slowly added dropwise thereto, the reaction was continued at 0 ℃ for 30 minutes after the addition, 1, 4-dibromobutane (83 mg,0.39 mmol) was slowly added dropwise thereto, the reaction was continued at 0 ℃ for 30 minutes after the addition, and TLC (PE/ea=4/1, r _f =0.3) was monitored for completion. The reaction mixture was quenched with saturated brine (20 mL), extracted with dichloromethane (20 mL x 3), the combined organic phases dried over anhydrous sodium sulfate, concentrated, and the crude product purified by silica gel column chromatography (PE/ea=5/1) to give 2g (30 mg, yield 24.23%) of the title compound as a colorless oil.

¹H NMR(400MHz,CDCl₃)δ7.81(s,2H),3.94(s,3H),3.93(s,3H),2.53-2.46(m,2H),2.08-1.94(m,6H).

Seventh step 3-bromo-5- (1-cyanocyclopentyl) -2-methoxybenzoic acid 2h

2G (2.94 g,8.69 mmol) of the compound was added to a mixed solution of methanol (15 mL) and water (5 mL), sodium hydroxide (452 mg,11.3 mmol) was added thereto, and the reaction was allowed to react at room temperature for 4 hours, and the reaction was completed as monitored by TLC (PE/EA=4/1, R _f =0.6). The reaction solution was concentrated under reduced pressure at 20℃to remove methanol from the reaction solution. To this was slowly added dropwise (1M/L) dilute hydrochloric acid to acidify to ph=4-5, extracted with ethyl acetate (20 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate, and the organic phase was concentrated to dryness to give the title compound as a white solid for 2h (2.81 g, 99%).

LC-MS:m/z=326.0[M+H]⁺。

Eighth step 3-bromo-5- (1-cyanocyclopentyl) -2-methoxy-N-methylbenzamide 2i

Compound 2h (2.81 g,8.67 mmol) was added to dichloromethane (30 mL), to which was added urea N, N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate (6.60 g,17.3 mmol), methylamine hydrochloride (877 mg,13.0 mmol) and N, N-diisopropylethylamine (4.47 g,34.6 mmol), the reaction was reacted at room temperature for 18 h, TLC detection (DCM/meoh=10/1, r _f =0.2) was complete, the reaction was quenched with water (20 mL), extracted with dichloromethane (50 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 (PE/ea=2/1) to give compound 2i (2.62 g, yield 89.73%) as a colourless oil.

Ninth step 5- (1-cyanocyclopentyl) -2-methoxy-3- (methylcarbamoyl) benzoic acid methyl ester 2j

Compound 2i (284 mg,0.842 mmol) was dissolved in a mixed solution of methanol (10 mL) and DMF (2 mL), transferred to an autoclave, and bis-triphenylphosphine palladium dichloride (100 mg,0.142 mmol) was added thereto, carbon monoxide was displaced 3 times, carbon monoxide was flushed therein, the pressure was maintained at 3-4 atmospheres, the reaction solution was heated to 90℃for 18 hours, a large amount of starting material remained by TLC monitoring (PE/EA=1/1, R _f =0.4), the reaction solution was concentrated to dryness, and the crude product was purified by silica gel column chromatography (PE/EA=1/1) to give the title compound 2j (21 mg, yield 7.88%) as a colorless oil.

LC-MS:m/z=317.2[M+H]⁺

Tenth step 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), sodium hydroxide (15 mg,0.37 mmol) was added thereto, and the reaction was continued at room temperature for 18 hours after the addition, and the reaction was completed by TLC monitoring (PE/ea=1/1, r _f =0.5). The reaction mixture was concentrated to remove methanol, quenched by adding water (20 mL), and slowly acidified by dropwise addition of (1M/L) dilute hydrochloric acid to ph=4-5. Ethyl acetate extraction (20 ml x 3), combining the organic phases, drying over anhydrous sodium sulfate, and concentration gave the title compound 2k (30 mg, crude) as a white solid, which was used directly in the next step.

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

Eleventh step N ¹ - ((5-bromothiophen-2-yl) sulfonyl) -5- (1-cyanocyclopentyl) -2-methoxy-N ³ -methyl isophthalamide 2L

Compound 2k (30 mg,0.099 mmol) was added to dichloromethane (5 mL), and to this was added nitrogen, nitrogen' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (60 mg,0.15 mmol) and N, N-diisopropylethylamine (0.1 mL,0.5 mmol), and the reaction was reacted at room temperature for 18 hours, after which the reaction was completed by TLC monitoring (DCM/meoh=10/1, r _f =0.2). The reaction mixture 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), 5-bromothiophene-2-sulfonamide (30 mg,0.12 mmol) and N, N-diisopropylethylamine (0.1 mL,0.5 mmol) were added sequentially and reacted at room temperature for 4 hours, TLC monitoring (DCM/meoh=20/1, r _f =0.4) was complete, the reaction quenched with water (5 mL), extracted with dichloromethane (10 mL x 2), the organic phases were combined, concentrated and the crude product purified by silica gel column chromatography (DCM/meoh=60/1) to give the title compound 2L as a white solid (30 mg, two step yield 30.00%).

LC-MS:m/z=526.0[M+H]⁺

Twelfth step N ¹ - ((5-bromothiophen-2-yl) sulfonyl) -5- (1-cyanocyclopentyl) -2-hydroxy-N ³ -methyl isophthalamide 2

Compound 2L (30 mg,0.056 mmol) was added to dichloromethane (4 mL), to which was added a boron tribromide-dichloromethane solution (2 mL,2mmol,1 m/L), and the reaction was allowed to react at room temperature for 1 hour, monitored by TLC (DCM/meoh=20/1, r _f =0.5) and complete. The reaction was quenched with water (10 mL), extracted with dichloromethane (5 mL x 3), the combined organic phases dried over anhydrous sodium sulfate, and concentrated, and the crude product purified by silica gel column chromatography (DCM/meoh=20/1) to give the title compound 2 (20 mg, yield 68.49%) as a white solid.

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

¹H NMR(400MHz,DMSO-d₆)δ15.98(s,1H),8.51(d,J＝4.4Hz,1H),8.02(dd,J＝6.8,2.8Hz,2H),7.41(d,J＝3.6Hz,1H),7.19(d,J＝4.0Hz,1H),2.83(d,J＝4.8Hz,3H),2.36(dd,J＝17.6,4.8Hz,2H),2.05-1.95(m,2H),1.91-1.84(m,4H).( Sulfonamide active hydrogens not shown).

Example 3

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

First step 2-chloro-4-methylthiazole-5-sulfonamide 3b

2-Chloro-4-methylthiazole-5-sulfonyl chloride 3a (160 mg,0.689 mmol) was dissolved in 1, 4-dioxane (1 mL), cooled to 0℃and ammonia (0.5 mL) was slowly added dropwise, and the reaction mixture was warmed to room temperature after the addition to react for 10 minutes, and TLC showed complete reaction of the starting materials. The reaction solution was directly concentrated to give the title compound 3b (169 mg, crude) as a white solid, which was used directly in the next step.

LC-MS:m/z=212.9[M+H]⁺

Second step 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 sequentially at room temperature, followed by a toluene solution (1 mL) of compound IN-1 (214 mg,1.02 mmol) and reacted overnight at room temperature after the addition, and TLC showed complete starting material reaction. The reaction mixture 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, concentrated, and the crude product was purified by Prep-TLC (ethyl acetate/petroleum ether=1/3) to give the title compound 3 as a white solid (60 mg, 23% yield in two steps).

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

¹H NMR(400MHz,DMSO-d₆)δ7.64(d,J＝2.0Hz,1H),7.56(d,J＝8.4Hz,1H),7.45(dd,J＝8.4,2.0Hz,1H),2.53(s,3H).( Sulfonamide active hydrogens not shown).

Example 4

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

First step 1-cyclopropyl-2-thiocyanate-1-one 4b

Compound A-bromo-cyclopropylethanone 4a (20.1 g,123 mmol) was dissolved in absolute ethanol (170 mL), sodium thiocyanate (12.1 g,149 mmol) was added at room temperature, the reaction was completed for 10 hours at room temperature, and TLC monitored for disappearance of starting material. Filtration, washing of the filter cake with anhydrous diethyl ether (50 ml x 2), concentration of the filtrate, dissolution of the crude anhydrous diethyl ether (50 ml x 2), filtration, concentration of the filtrate, and direct proceeding to the next step gave the title compound 4b (10.2 g, crude) as a dark brown liquid.

Second step 2-chloro-4-cyclopropylthiazole 4c

Compound 4b (10.2 g, crude) was dissolved in dichloromethane (120 mL) and dried hydrogen chloride gas was slowly vented at 0 ℃ for 1 hour, slowly warmed to room temperature for further reaction for 10 hours, TLC showed the disappearance of starting material. The reaction mixture was quenched with water (200 mL), extracted with dichloromethane (60 mL x 3), the organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the crude product purified by silica gel column chromatography (ethyl acetate/petroleum ether=1/5) to give the title compound 4c (4.11 g, two-step yield 20.4%)

LCMS:m/z=160.1[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ6.73(s,1H),1.96(tt,J=8.2,5.2Hz,1H),0.95-0.86(m,4H).

Third step 2-chloro-4-cyclopropylthiazole-5-sulfonyl chloride 4d

Compound 4C (300 mg,1.88 mmol) was dissolved in anhydrous tetrahydrofuran (7 mL) under nitrogen, cooled to-70℃and n-butyllithium (2.5M in n-hexane, 1 mL) was slowly added dropwise with a small warming. The reaction was continued for 10 minutes at-70 ℃. And introducing dry sulfur dioxide gas for 20 minutes, continuing the reaction for 10 minutes, and returning to room temperature for reaction for 1 hour. The reaction was concentrated, the residue was dissolved in dichloromethane (3 mL), N-chlorosuccinimide (376 mg,2.82 mmol) was added and reacted at room temperature for 10 hours, and TLC showed the disappearance of starting material. The reaction 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 give the title compound 4d (322 mg, crude) as a colorless oil, which was used directly in the next step.

Fourth step 2-chloro-4-cyclopropylthiazole-5-sulfonamide 4e

Compound 4d (322 mg, crude) was dissolved in 1, 4-dioxane (2 mL), ammonia (25%, 0.5 mL) was added at room temperature, and after addition, reaction was performed at room temperature for 10 min, TLC showed the disappearance of starting material. The reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with saturated brine (30 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 give the title compound 4e (178 mg, 40% yield in two steps) as a white solid

LCMS:m/z=238.9[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ5.10(s,2H),2.52(dq,J=8.0,5.2Hz,1H),1.18-1.08(m,4H).

Fifth step 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), HATU (239 mg,0.63 mmol) and N, N-diisopropylethylamine (108 mg,0.84 mmol) were added at room temperature, and after the addition, the reaction was continued at room temperature for 2 hours and TLC showed the disappearance of starting material. The reaction was quenched with water (2 mL), dried over anhydrous sodium sulfate, concentrated, and the crude was dissolved in dichloromethane (2 mL), compound 4e (100 mg,0.42 mmol) was added and reacted at room temperature for 10 hours, TLC showed the starting 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, concentrated, and the crude product was purified by Pre-HPLC (acetonitrile/water=60/40) to give the title compound 4 (35 mg, yield 20%) as a white solid

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

¹H NMR(400MHz,CDCl₃)δ9.20(s,1H),7.77(d,J＝8.4Hz,1H),7.48(d,J＝2.0Hz,1H),7.39(dd,J＝8.8,2.0Hz,1H),2.72-2.65(m,1H),1.21-1.13(m,4H).( Sulfonamide active hydrogens not shown).

Example 5

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

First step 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 0deg.C, N-dimethylformamide (4 drops) was added, and after completion of the reaction at 0deg.C for 2 minutes oxalyl chloride (2 mL,23.6 mmol) was slowly added dropwise, and the addition was warmed to room temperature for 4 hours, and TLC showed disappearance of starting material. The reaction solution was concentrated to dryness to give the title compound 5b (230 mg, crude) as a white solid, which was used directly in the next step.

Second step 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), 4-dimethylaminopyridine (5 mg,0.04 mmol), compound 5b (230 mg, crude product) and triethylamine (2 mL,14.4 mmol) were added sequentially at room temperature, and after addition, the reaction was continued at room temperature for 2 hours, and TLC showed the disappearance of starting material. The reaction mixture was quenched with water (10 mL), extracted with ethyl acetate (10 mL. Times.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 give 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)

¹H NMR(400MHz,DMSO-d₆)δ7.99(s,1H),7.78(q,J＝8.4Hz,2H),7.70(d,J＝4.0Hz,1H),7.42(d,J＝4.4Hz,1H).( Sulfonamide active hydrogens not shown)

Example 6

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

First step 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), urotropine (4.86 g,34.71 mmol) and cuprous oxide (2.48 g,17.3 mmol) were added at room temperature, and the reaction mixture was heated to 80℃for 10 hours after the addition, and TLC showed the disappearance of starting material. The reaction solution was cooled to room temperature, hydrochloric acid (3 n,70 mL) was slowly added dropwise, and after the addition, the reaction was continued at room temperature for 2 hours, concentrated, the remainder was added to water (100 mL), filtered, and the cake was rinsed again with water (300 mL), and dried to give the title compound 6b (7.02 g, crude product) as a white solid, which was used directly in the next step.

Second step 4-chloro-3-formyl-2-hydroxybenzoic acid methyl ester 6c

Compound 6b (7.02 g, crude) was added to anhydrous methanol (70 mL), concentrated sulfuric acid (7 mL) was added at room temperature, and the reaction mixture was heated to 80℃for 10 hours after the addition, and TLC showed the disappearance of starting material. The reaction solution was cooled to room temperature, quenched with water (100 mL), extracted with ethyl acetate (40 ml×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 give the title compound 6c (3.47 g, two-step yield 93%)

Third step 4-chloro-3-formyl-2- (methoxymethoxy) benzoic acid methyl ester 6d

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

Fourth step 6-chloro-2-hydroxy-3- (methoxycarbonyl) benzoic acid 6e

Compound 6d (1.00 g,3.86 mmol) was added to acetonitrile (20 mL), 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, and after the addition, the reaction was continued at room temperature for 10 hours, and TLC showed the disappearance of starting material. The reaction solution was added with saturated aqueous sodium sulfite solution (40 mL), at least a part of the solvent was concentrated, hydrochloric acid (3N) was slowly added dropwise to ph=1-2, stirred for 2 hours, and then filtered, and the filter cake was dried to give the title compound 6e (500 mg, yield 56%) as a white solid.

Fifth step 4-chloro-2-hydroxy-3- (methylcarbamoyl) benzoic acid methyl ester 6f

Compound 6e (227 mg,2.28 mmol) was added to dichloromethane (20 mL), and methyl amine 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 and reacted at room temperature for 10 hours, TLC showed the starting material disappeared. The reaction mixture 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 give the title compound 6f (450 mg, crude) as an off-white solid, which was used directly in the next step.

Sixth step 6g of 4-chloro-2-hydroxy-3- (methylcarbamoyl) benzoic acid

Compound 6f (310 mg, crude) was dissolved in tetrahydrofuran (6 mL), aqueous sodium hydroxide (15%, 1 mL) was added at room temperature, and the reaction was continued at room temperature for 10 hours after the addition, and TLC showed the disappearance of starting material. The reaction solution was quenched with water (6 mL), acidified with dilute hydrochloric acid (3N) to ph=3, concentrated, filtered, and the filter cake was washed with water (5 ml×2) and dried to give the title compound 6g (190 mg, two step yield 52%)

Seventh step N ¹ - ((5-bromothiophen-2-yl) sulfonyl) -4-chloro-2-hydroxy-N3-methyl isophthalamide 6

6G (190 mg,0.83 mmol) of the compound was dissolved in dichloromethane (4 mL), N-diisopropylethylamine (214 mg,1.65 mmol), HATU (470 mg,1.24 mmol) and 5-bromo-2-thiophenesulfonamide (220 mg,0.91 mmol) were added in this order at room temperature, and after the addition, the reaction was continued at room temperature for 3 hours and TLC showed the disappearance of starting material. The reaction mixture was quenched with water (2 mL), extracted with dichloromethane (1 mL. Times.3), the organic phases were combined, washed with saturated brine (1 mL), and the organic phase was concentrated to give the title compound 6 (4.51 mg, yield 1.2%) as a white solid

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

¹H NMR(400MHz,CDCl₃)δ11.02(s,1H),8.14(d,J＝8.8Hz,1H),7.84(s,1H),7.71(d,J＝4.0Hz,1H),7.09(d,J＝4.0Hz,1H),7.03(d,J＝8.8Hz,1H),3.09(d,J＝4.8Hz,3H).( Sulfonamide active hydrogens not shown)

Example 7

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

First step 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 ℃, a dilution of chlorosulfonic acid (3 mL) in dichloromethane (3 mL) was slowly added, and the reaction was continued for 10min at 0 ℃ with TLC indicating the disappearance of starting material. The reaction was slowly poured into ice water (20 mL) to quench, extracted with dichloromethane (6 ml×3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 7b (916 mg, crude) as a colourless oil, which was used directly in the next step.

Second step 3-bromo-5-methylthiophene-2-sulfonamide 7c

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

Third step N- ((3-bromo-5-methylthiophene-2-yl) sulfonyl) -2, 4-dichlorobenzamide 7

Compound 7C (293 mg, crude) was dissolved IN ethyl acetate (2 mL), triethylamine (383 mg,3.82 mmol) was added, reacted at room temperature for 10 min, cooled to 0 ℃, intermediate IN-1 (200 mg,0.95 mmol) IN toluene (1 mL) was added and reacted at room temperature for 10h, TLC showed the disappearance of starting material. The reaction was quenched with water (20 mL), extracted with ethyl acetate (8 mL x 3), the organic phases combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the crude product purified by Prep-TLC (dichloromethane/methanol=10/1) to give the title compound 7 (110 mg, three step yield 35%) as a white solid.

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

¹H NMR(400MHz,DMSO-d₆ ) Delta 7.73 (d, j=1.2 hz, 1H), 7.57-7.50 (m, 2H), 7.10 (s, 1H), 2.52 (s, 3H), (sulfonamide active hydrogen not shown).

Example 8

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

First step 3-chloro-4-hydroxy-5-methoxybenzaldehyde 8b

Vanillin 8a (15.1 g,99.3 mmol) was added to N, N-dimethylformamide (100 mL), N-chlorosuccinimide (19.9 g,149 mmol) was added at room temperature, and the reaction was continued at room temperature for 10 hours, with TLC showing the disappearance of starting material. The reaction mixture 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 purified by silica gel column chromatography (ethyl acetate/petroleum ether=1/15) to give the title compound 8b (8.22 g, yield 44%) as a white solid, which was used directly in the next step.

Second step 3-chloro-4-trifluoromethanesulfonyl-5-methoxybenzaldehyde 8c

Compound 8b (8.22 g,44.0 mmol) was added to dichloromethane (150 mL), triethylamine (13.4 mg,132 mmol) and 4-dimethylaminopyridine (279 mg,2.20 mmol) were added at room temperature, the reaction solution was cooled to 0℃after the addition, N-phenylbis (trifluoromethanesulfonyl) imine solution (20.5 g,57.3mmol, dissolved in 40mL of dichloromethane) was added dropwise, and the reaction was continued at room temperature for 10 hours with TLC showing the disappearance of starting material. The reaction was quenched with water (200 mL), extracted with dichloromethane (50 mL x 3), the organic phases combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 8c (15.3 g, crude) as an off-white solid, which was used directly in the next step.

Third step 2-chloro-4-formyl-6-methoxybenzoic acid methyl ester 8d

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

Fourth step 2-chloro-4- (hydroxymethyl) -6-methoxybenzoic acid methyl ester 8e

Compound 8d (8.49 g,37.1 mmol) was added to methanol (10 mL), sodium borohydride (702 mg,18.6 mmol) was added at room temperature, and the reaction was completed at room temperature for 30min, with TLC indicating the disappearance of starting material. The reaction was quenched with water (80 mL), extracted with ethyl acetate (30 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 8e (6.81 g, crude) as a pale yellow oil, which was used directly in the next step.

Fifth step methyl 2-chloro-6-methoxy-4- (((methylsulfonyl) oxy) methyl) benzoate 8f

Compound 8e (6.81 g, crude) was added to dichloromethane (50 mL), cooled to 0deg.C, triethylamine (6.01 g,59.1 mmol) was added, stirred for 10min, methylsulfonyl chloride (4.06 g,35.4 mmol) was added, and the addition was warmed to room temperature for 30 min, and TLC showed the starting material disappeared. The reaction 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 give the title compound 8f (7.26 g, crude) as a colorless oil, which was used directly in the next step.

Sixth step 8g of 2-chloro-4- (cyanomethyl) -6-methoxybenzoic acid methyl ester

Compound 8f (7.26 g, crude) was added to acetonitrile (40 mL), anhydrous potassium carbonate (3.90 g,28.2 mmol) and trimethylcyanosilane (3.50 g,35.2 mmol) were added sequentially at room temperature, and the addition was warmed to 50deg.C after 12 hours of reaction, and TLC showed the disappearance of starting material. The reaction mixture 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, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether=1/5) to give 8g (1.81 g, three-step yield 20%) of the title compound as a colorless oil.

Seventh step 2-chloro-4- (1-cyanocyclopropyl) -6-methoxybenzoic acid methyl ester 8h

8G (1.22 g,5.09 mmol) of the compound was added to tetrahydrofuran (10 mL), cooled to 0℃and potassium tert-butoxide (1.26 mg,11.2 mmol) was added slowly in portions, reacted at 0℃for 30 minutes, 1, 2-dibromoethane (956 mg,5.09 mmol) was added and the addition was warmed to room temperature for 30 minutes, and TLC showed the disappearance of starting material. The reaction mixture was quenched with water (10 mL), extracted with ethyl acetate (5 ml×3), the organic phases were combined, washed with saturated brine (8 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 give the title compound as a white solid 8h (224 mg, yield 16%).

Eighth step 2-chloro-4- (1-cyanocyclopropyl) -6-methoxybenzoic acid 8i

Compound 8h (224 mg,0.84 mmol) was dissolved in anhydrous methanol (2 mL), sodium hydroxide solution (15%, 1 mL) and water (0.5 mL) were added at room temperature, and after the addition, the temperature was raised to 80℃for 10 hours, and TLC showed the disappearance of starting material. The reaction solution was cooled naturally to room temperature, acidified to ph=7 with hydrochloric acid (3N), concentrated to remove methanol, 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×2) and dried to give the title compound 8i (110 mg, yield 52%) as a white solid.

Ninth step 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-diisopropylethylamine (113 mg,0.87 mmol) and 5-bromo-2-thiophenesulfonamide (106 mg,0.44 mmol) were added sequentially at room temperature. The reaction was carried out at room temperature for 10 hours, and TLC showed the disappearance of starting material. The reaction mixture 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 give the title compound 8 (33 mg, 16% yield) as a white solid

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

¹H NMR(400MHz,DMSO-d₆)δ7.64(d,J＝3.6Hz,1H),7.42(d,J＝4.0Hz,1H),7.03(d,J＝1.2Hz,1H),6.90(s,1H),3.79(s,3H),1.79(dd,J＝8.4,5.2Hz,2H),1.63(dd,J＝8.0,4.8Hz,2H).( Sulfonamide active hydrogens not shown).

Example 9

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

First step N- ((5-bromothiophen-2-yl) sulfonyl) -2-chloro-4- (1-cyanocyclopropyl) -6-hydroxybenzoamide 9

Compound 8 (20 mg,42 mmol) was dissolved in dichloromethane (1 mL), a solution of boron tribromide in dichloromethane (1 mL,1mmol, 1N) was added at room temperature, and the reaction was completed at room temperature for 30 min, and TLC showed the disappearance of starting material. The reaction mixture 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, concentrated, and the crude product was purified by Prep-HPLC to give the title compound 9 (8.51 mg, 43% yield) as a white solid.

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

¹H NMR(400MHz,DMSO-d₆)δ13.78(s,1H),7.38(s,1H),7.19(s,1H),6.70(d,J＝2.0Hz,1H),6.68(s,1H),1.74(dd,J＝8.4,5.2Hz,2H),1.55(dd,J＝8.0,5.2Hz,2H).( Sulfonamide active hydrogens not shown).

Example 10

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

First step (3R, 4S, 5R) -3,4, 5-Triacetoxycyclohex-1-en-1-carboxylic acid 10b

(3R, 4S, 5R) -3,4, 5-triacetoxy-cyclohex-1-ene-1-carboxylic acid 10a (1.02 g,5.74 mmol) was dissolved in methylene chloride (10 mL), triethylamine (1.16 g,11.5 mmol) was added at room temperature, the reaction was completed at room temperature for 10 minutes, acetic anhydride (2.34 g,23.0 mmol) was slowly added dropwise to the reaction solution after cooling to 0℃and the reaction solution was warmed up slightly. After addition, reaction was carried out at room temperature for 30min, TLC showed disappearance of starting material. The reaction was quenched with water (10 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 to give the title compound 10b (1.21 g, crude) as a colorless oil, which was used directly in the next step.

Second step (1R, 2S, 3R) -5- (Chlorocarbonyl) cyclohex-4-en-1, 2, 3-triacetate triester 10c

Compound 10b (1.21 g, crude product) was dissolved in tetrahydrofuran (10 mL), cooled to 0deg.C, oxalyl chloride (1.42 g,11.2 mmol) was slowly added dropwise, reacted at 0deg.C for 10 min, followed by N, N-dimethylformamide (3 drops) slowly added dropwise, the reaction was vigorous with a large amount of gas generation, warmed to room temperature after addition, reacted for 30 min, and TLC monitored the completion of the starting material reaction. The reaction solution was concentrated to give the title compound 10c (725 mg, crude) as a white solid, which was used directly in the next step.

Third step (1R, 2S, 3R) -5- (((5-bromothiophen-2-yl) sulfonyl) carbamoyl) cyclohex-4-en-1, 2, 3-triacetate 10d

5-Bromothiophene-2-sulfonamide (706 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, the reaction was completed for 10min at room temperature, a toluene (2 mL) solution of compound 10c (725 mg,2.27 mmol) was added, and the reaction was continued at room temperature for 12 hours, TLC showed the starting material to disappear. The reaction mixture was quenched with water (5 mL), extracted with ethyl acetate (2 mL. Times.3), the organic phases were combined, washed with saturated brine (3 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 10d (1.20 g, crude) as a colorless oil, which was used directly in the next step.

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

Compound 10d (1.20 g, crude) was dissolved in anhydrous methanol (10 ml), anhydrous potassium carbonate (1.26 g,9.15 mmol) was added at room temperature, and the reaction was completed at room temperature for 2 hours, and TLC showed the disappearance of starting material. The reaction 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 give the title compound 10 as a white solid (47 mg, four step yield 2%).

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

¹H NMR(400MHz,CD₃OD)δ7.52(d,J＝4.0Hz,1H),7.11(d,J＝4.4Hz,1H),6.46-6.45(m,1H),4.28(s,1H),3.88(dt,J＝6.7,4.8Hz,1H),3.60(dd,J＝6.8,4.0Hz,1H),2.58-2.52(m,1H),2.02(dd,J＝18.0,4.8Hz,1H).( Sulfonamide active hydrogens not shown).

Example 11

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

First step (Z) -N' - ((5-bromothiophen-2-yl) sulfonyl) -N, N-dimethylformamide 11b

5-Bromothiophene-2-sulfonamide 11a (2.40 g,9.92 mmol) was added to dichloromethane (10 mL), N-dimethylformamide dimethyl acetal (2 mL) was added at room temperature, and after addition, the reaction was continued at room temperature for 20min, and TLC showed the disappearance of starting material. The reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 11b (3.01 g, crude) as a pale yellow solid, which was used directly in the next step.

Second step (Z) -N, N-dimethyl-N' - ((5- ((trimethylsilyl) ethynyl) 2-thienyl) sulfonyl) imide 11c

Compound 11b (2.00 g, 6.284 mmol) was added to dry N, N-dimethylformamide (20 mL), triethylamine (4.70 mL,33.8 mmol), ketiden iodide (130 mg,0.68 mmol), bis-triphenylphosphine palladium dichloride (240 mg,0.34 mmol) and trimethylethynyl silicon (1.35 mL,9.46 mmol) were added sequentially at room temperature, the addition was completed, nitrogen was displaced 3 times, and the reaction was continued at room temperature for 24 hours, and TLC showed the disappearance of starting material. The reaction was filtered, the filter cake was washed with ethyl acetate (10 mL x 2), the filtrate was diluted with water (100 mL), ethyl acetate (30 mL x 3) was extracted, the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by Prep-TLC (petroleum ether/ethyl acetate=10/1) to give the title compound 11c (1.15 g, 37% yield in two steps) as a pale yellow solid.

Third step 5-Acetylthiophene-2-sulfonamide 11d

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

Fourth step 2, 4-dichloro-N- ((5-ethynyl thiophen-2-yl) sulfonyl) benzamide 11

2, 4-Dichlorobenzoic acid (204 mg,1.07 mmol) was dissolved in dichloromethane (4 mL), HATU (319 mg,1.60 mmol) and N, N-diisopropylethylamine (276 mg,2.14 mmol) were added at room temperature, and after addition the reaction was completed at room temperature for 2 hours, TLC showed the disappearance of starting material. The reaction mixture was quenched with water (4 mL), extracted with dichloromethane (2 mL x 3), the organic phases combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, concentrated, the residue dissolved in dichloromethane (4 mL), N-diisopropylethylamine (276 mg,2.14 mmol) and compound 11d (200 mg,1.61 mmol) were added sequentially at room temperature, and reacted at room temperature for 10 hours, TLC showed the disappearance of starting material. The reaction mixture was quenched with water (4 mL), extracted with dichloromethane (2 mL x 3), the organic phases combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product purified by Prep-HPLC to give the title compound 11 as a white solid (75 mg, 19% yield).

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

¹H NMR(400MHz,CDCl₃)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-iodothiophen-2-yl) sulfonyl) benzamide 12

First step 5-iodothiophene-2-sulfonyl chloride 12b

2-Iodothiophene 12a (4.01 g,19.0 mmol) was added to dichloromethane (20 mL), cooled to-60℃and chlorosulfonic acid (5 mL) was slowly added dropwise, reacted at-60℃for 10 min, warmed to room temperature for 2h, and TLC showed the disappearance of starting material. The reaction was quenched by addition of ice water (50 mL), extracted with dichloromethane (20 mL x 3), the organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, and concentrated to afford the title compound 12b (3.02 g, crude) as a pale yellow oil, which was used directly in the next step.

Second step 5-iodothiophene-2-sulfonamide 12c

Compound 12b (3.02 g, crude) was added to 1, 4-dioxane (10 mL), ammonia (2 mL) was added at room temperature, and the reaction was completed at room temperature for 20 min, with TLC indicating the disappearance of starting material. The reaction was quenched by pouring into water (20 mL), extracting with dichloromethane (10 mL. Times.3), combining the organic phases, washing with saturated brine (10 mL), drying over anhydrous sodium sulfate, and concentrating to give the title compound 12c (3.35 g, crude) as a pale yellow oil, which was used directly in the next step.

Third step 2, 4-dichloro-N- ((5-iodothiophen-2-yl) sulfonyl) benzamide 12

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

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

¹H NMR(400MHz,DMSO-d₆)δ7.72(d,J＝2.0Hz,1H),7.55(d,J＝8.4Hz,1H),7.51(dd,J＝4.0,3.2Hz,2H),7.48(d,J＝3.6Hz,1H).( Sulfonamide active hydrogens not shown).

Example 13

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

First step 6-bromo-N- ((5-bromothiophen-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), 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 sequentially at room temperature and reacted at room temperature for 12 hours, TLC showed the starting material disappeared. The reaction mixture was quenched with water (2 mL), extracted with ethyl acetate (2 ml×3), the organic phases combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product purified by Prep-TLC (ethyl acetate/petroleum ether=1/8) to give the title compound 13 (48 mg, yield 28%) as a white solid

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

¹H NMR(400MHz,DMSO-d₆)δ:8.22(d,J＝1.6Hz,1H),7.84-7.77(m,2H),7.50(dd,J＝1.6,2.0Hz,1H),7.33(d,J＝4.0Hz,1H),7.14(d,J＝4.0Hz,1H).( Sulfonamide active hydrogens not shown).

Example 14

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

First step 2, 4-dichloro-N- ((4, 5-dichlorothiophen-2-yl) sulfonyl) benzamide 14

Intermediate IN-1 (185 mg, crude) 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 sequentially at room temperature and reacted at room temperature for 1 hour. TLC detection of completion of the reaction starting material, quenching of the reaction solution with water (20 mL), extraction of dichloromethane (10 mL x 3), combining organic phases, drying over anhydrous sodium sulfate, concentration, purification of the crude product by Prep-HPLC gave the title compound 14 (65 mg, yield 19.34%) as a white solid.

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

¹H NMR(400MHz,DMSO-d₆)δ7.88(s,1H),7.71(d,J＝2.0Hz,1H),7.61(d,J＝8.4Hz,1H),7.50(dd,J＝8.4,2.0Hz,1H).( Sulfonamide active hydrogens not shown).

Example 15

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

First step 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), triethylamine (3.50 g,34.6 mmol) was added at room temperature, the reaction solution cooled to-60℃after the addition, bromine (5.01 g,31.3mmol, dissolved in 20mL dichloromethane) was slowly added dropwise, and the reaction was continued for 1 hour at-60℃until the addition was complete, and TLC showed the disappearance of starting material. 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 (1 n,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 give the title compound 15b (2.20 g, yield 28%)

Second step 5-bromo-4-chloro-2-hydroxybenzoic acid ethyl ester 15c

Compound 15b (1.00 g,3.98 mmol) was dissolved in absolute ethanol (20 mL), concentrated sulfuric acid (2 mL) was added at room temperature, and the temperature was raised to 80℃under nitrogen protection for 10 hours, and TLC showed the disappearance of starting material. The reaction 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 give the title compound 15c (1.02 g, yield 92%) as a colorless oil.

Third step 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), 2-cyanobenzeneboronic acid (566 mg,3.85 mmol), potassium phosphate (1.16 mg,5.47 mmol) and Pd (dppf) ₂Cl₂ (200 mg,0.27 mmol) were added sequentially at room temperature, and after the addition, the reaction was warmed to 90℃and allowed to react for 2 hours, and TLC showed the disappearance of starting material. The reaction mixture 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 give the title compound 15d as an off-white solid (230 mg, yield 28%).

Fourth step 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), sodium hydroxide solid (50 mg,1.25 mmol) was added at room temperature, and the reaction mixture was reacted at room temperature for 12 hours after the addition, and TLC showed the disappearance of starting material. The reaction mixture 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 give the title compound 15e as a white solid (85 mg, yield 76%).

Fifth step 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), HATU (177 mg,0.47 mmol), N-diisopropylethylamine (80 mg,0.62 mmol) and 5-bromo-2-thiophenesulfonamide (75 mg,0.31 mmol) were added sequentially at room temperature, and the reaction solution was reacted at room temperature for 12 hours. The reaction mixture 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 purified by Prep-HPLC to give the title compound 15 (40 mg, 26% yield) as a white solid

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

¹H NMR(400MHz,CDCl₃)δ7.76(d,J＝7.6Hz,1H),7.70-7.66(m,2H),7.55-7.51(m,2H),7.42(d,J＝8.0Hz,1H),7.19(s,1H),7.11(d,J＝3.6Hz,1H).( Sulfonamide and phenolic hydroxyl active hydrogens are not shown).

Example 16

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

First step 2-chloro-4- (1-cyanocyclopentyl) -6-methoxybenzoic acid methyl ester 16a

8G (1.13 g,4.71 mmol) of the compound was added to tetrahydrofuran (20 mL), cooled to 0℃and potassium tert-butoxide (1.17 g,10.4 mmol) was added slowly in portions, after 30 minutes of reaction at 0℃1, 4-dibromobutane (1.02 g,4.73 mmol) was added, the reaction mixture was warmed slowly to room temperature and reacted for 30 minutes, and TLC showed the disappearance of starting material. The reaction mixture was quenched with water (30 mL), extracted with ethyl acetate (15 ml×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=1/5) to give the title compound 16a as a white solid (1.02 g, yield 74%).

Second step 2-chloro-4- (1-cyanocyclopentyl) -6-methoxybenzoic acid 16b

Compound 16a (200 mg,0.68 mmol) was dissolved in anhydrous methanol (6 mL), sodium hydroxide solution (15%, 1 mL) and water (1 mL) were added at room temperature, and the reaction mixture was warmed to 80℃for 10 hours after the addition, and TLC showed the disappearance of starting material. The reaction solution was cooled naturally to room temperature, diluted hydrochloric acid (3N) was added to acidify the reaction solution to ph=7, methanol was concentrated to minimum volume, the remainder was diluted with water (1 mL), hydrochloric acid (3N) was acidized to ph=1, a large amount of white solid was precipitated, filtered, and the cake was washed with water (1 mL x 2) and dried to give the title compound 16b (165 mg, yield 87%) as a white solid.

Third step 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-diisopropylethylamine (152 mg,1.18 mmol) and 5-bromo-2-thiophenesulfonamide (143 mg, 0.560 mmol) were added sequentially at room temperature and reacted for 10 hours at room temperature, TLC showed the starting material disappeared. The reaction mixture 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 purified by Prep-HPLC to give the title compound 16 (60 mg, 20.19% yield) as a white solid

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

¹H NMR(400MHz,CDCl₃)δ8.44(s,1H),7.71(d,J＝4.0Hz,1H),7.13(d,J＝4.0Hz,1H),7.04(d,J＝1.2Hz,1H),6.96(d,J＝1.2Hz,1H),3.86(s,3H),2.47(d,J＝5.2Hz,2H),2.10-1.93(m,6H).

Example 17

N- ((5-bromothiophen-2-yl) sulfonyl) -2-chloro-4- (1-cyanocyclopentyl) -6-hydroxybenzoamide 17

First step N- ((5-bromothiophen-2-yl) sulfonyl) -2-chloro-4- (1-cyanocyclopentyl) -6-methoxybenzamide 17

Compound 16 (20 mg,40 mmol) was dissolved in dichloromethane (1 mL), a solution of boron tribromide in dichloromethane (1 mL,1mmol, 1N) was added at room temperature, and after addition, reaction was performed at room temperature for 30 min, and TLC showed the disappearance of starting material. The reaction mixture 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, concentrated, and the crude product was purified by Prep-HPLC to give the title compound 17 (10 mg, 53% yield) as a white solid

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

¹H NMR(400MHz,CDCl₃)δ11.77(s,1H),10.04(s,1H),7.74(d,J＝4.0Hz,1H),7.14(d,J＝4.0Hz,1H),7.07(d,J＝2.0Hz,1H),7.03(d,J＝2.0Hz,1H),2.50-2.40(m,2H),2.07-1.95(m,6H).

Example 18

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

First 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) maintaining the drop temperature below 30 ℃. After the completion of the addition, the reaction was allowed to warm to 120℃for 24 hours, the reaction mixture was cooled to room temperature, thionyl chloride (60 mL) was added, and after the completion of the addition, the reaction was allowed to warm to 120℃for 24 hours, and TLC detected complete reaction of the starting materials (DCM/MeOH=20/1, R _f =0.2). The reaction solution was cooled to 0-10 ℃. Quench slowly dropwise in ice water (200 mL), extract with dichloromethane (50 mL x 5), combine the organic phases, dry over anhydrous sodium sulfate, concentrate, and purify the crude by silica gel column chromatography (ea=100%) to give the title compound 18b (15.02 g,2 points of tlc detection unable to be separated) as a yellow oil

Second step 1-ethyl-1H-imidazole-4-sulfonamide 18c

25% -28% Ammonia water (100 mL) is cooled to minus 15 ℃, tetrahydrofuran solution (15.02 g, crude product, dissolved in tetrahydrofuran 50 mL) of compound 18b is added dropwise, the dropwise adding temperature is kept below-10 ℃, and after the dropwise adding, TLC detection (DCM/MeOH=20/1, R _f =0.4) is completed. The reaction was concentrated and the crude product was purified by column chromatography on silica gel (DCM/meoh=10/1) to give 6.52g of crude product, which was heated to reflux with ethyl acetate (65 mL) and slurried for 30min, cooled naturally to room temperature, filtered, the ethyl acetate (20 mL) was rinsed with solids and the grey solids dried to give 3.10g of impurity in the product of ¹ H NMR analysis. The crude product was purified again by column chromatography on silica gel (DCM/meoh=15/1) to give 2.01g, which was heated to reflux with ethyl acetate (20 mL) and slurried for 30min, cooled naturally to room temperature, filtered, rinsed with ethyl acetate (10 mL) and dried to give the title compound 18c as an off-white solid (1.60 g, two step yield 4.3%).

¹H NMR(400MHz,DMSO-d₆)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)

Third step 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 mixture was warmed to 60 ℃ and reacted for 2 hours, the reaction mixture was naturally cooled to room temperature, compound 18C (352 mg,2.00 mmol) was added, after stirring the reaction mixture at room temperature for 10 minutes, 1, 8-diazabicyclo [5.4.0] undec-7-ene (300 mg,1.97 mmol) was added, stirring was continued at room temperature for 18 hours, TLC detection (DCM/MeOH=20/1, R _f =0.2) was complete, the reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (20 mL×2), and the aqueous phase was collected. Dilute hydrochloric acid (3M/L) was slowly added dropwise to acidify the aqueous phase to ph=4-5. Ethyl acetate (40 ml x 2) and the organic phases were combined. Drying over anhydrous sodium sulfate, concentrating, adding ethyl acetate (20 mL) into the crude product, heating to reflux, pulping for 0.5h, naturally cooling to room temperature, filtering, leaching the solid with ethyl acetate (10 mL), and drying 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)

¹H NMR(400MHz,DMSO-d₆)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).

Example 19

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

First step 1, 4-oxazane-4-sulfonamide 19b

1, 4-Oxazepine 19a (400 mg,3.95 mmol) was dissolved in 1, 4-dioxane (6 mL), sulfonamide (458 mg,4.74 mmol) was added at room temperature, and the reaction mixture was warmed to 120℃for 18 hours. TLC detection (DCM/meoh=20/1, r _f =0.1) starting material reacted completely. Naturally cooling the reaction solution to room temperature, and concentrating to dryness. Ethyl acetate (10 mL) was added to the residue and heated to reflux until complete dissolution, filtration was carried out while hot, the filtrate was naturally cooled to room temperature, a large amount of solids precipitated, filtration was carried out, the filter cake was rinsed with ethyl acetate (10 mL), and the filter cake was dried to give the title compound 19b (250 mg, yield 35.11%) as a yellow solid.

Second step N- (1, 4-oxazacyclo-4-ylsulfonyl) -2, 4-dichlorobenzamide 19

2, 4-Dichlorobenzoic acid (300 mg,1.57 mmol) was added to dichloromethane (10 mL), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (900 mg,2.36 mmol) and N, N-diisopropylethylamine (1 mL,5.42 mmol) were added sequentially at room temperature, and the reaction was continued for 18 hours. TLC detection (DCM/meoh=20/1, r _f =0.2) starting material reacted completely. The reaction mixture 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), compound 19b (250 mg,1.38 mmol) and N, N-diisopropylethylamine (1 mL,5.42 mmol) were added sequentially at room temperature, the reaction was continued for 18 hours after the addition, TLC detection (DCM/meoh=20/1, r _f =0.5) was complete, the reaction was quenched with water (20 mL), extracted with dichloromethane (20 ml×2) and the organic phases were combined. Dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/meoh=80/1) to give 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)

¹H NMR(400MHz,DMSO-d₆)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).

Example 20

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

First step 3-bromo-4-fluorobenzenesulfonamide 20b

3-Bromo-4-fluorobenzenesulfonyl chloride 20a (235 mg,0.859 mmol) was dissolved in1, 4-dioxane (4 mL), 25% -28% aqueous ammonia (0.5 mL) was slowly added dropwise, and after addition, the reaction was completed at room temperature for 30 minutes, and TLC detection (DCM/meoh=20/1, r _f =0.8) starting material was complete. The reaction was quenched with water (20 mL), extracted with ethyl acetate (15 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give the title compound 20b (229 mg, crude) as a white solid, which was used directly in the next step.

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

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

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

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

¹H NMR(400MHz,CDCl₃)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).

Example 21

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

First step 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 added slowly. After the addition, the reaction solution was heated to 90℃and reacted for 5 hours. TLC detection (PE/ea=10/1, r _f =0.4) starting material reacted completely. The reaction was cooled to room temperature, quenched with ice-water (500 mL), extracted with ethyl acetate (300 mL. Times.3), and the organic phases 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 give the title compound 21b (13.55 g, yield 93.12%) as a yellow solid.

¹H NMR(400MHz,CDCl₃)δ8.23(dd,J=6.4,2.4Hz,1H),6.25(s,2H).

Second step 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 added slowly. After the addition, the reaction solution was heated to 55℃and reacted for 18 hours. TLC detection (PE/ea=4/1, r _f =0.5) starting material reacted completely and the reaction was cooled to room temperature. Quench with water (100 mL), extract with ethyl acetate (200 mL x 3), and combine the organic phases. Dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (pe=100%) to give the title compound 21c (10.38 g, yield 87.96%) as a yellow oil.

Third step 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 sequentially at room temperature. After the addition, the reaction mixture was raised to 80 ℃ and reacted for 18 hours, and TLC detection (PE/ea=4/1, r _f =0.8) was complete. The reaction was cooled to room temperature, filtered, the solids rinsed with ethanol (100 ml x 3) and the filtrate concentrated. The residue was diluted with water (200 mL), extracted with ethyl acetate (200 mL x 3), and the organic phases combined. Dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (PE/ea=7/1) to give the title compound 21d (4.80 g, yield 52.9%) as a yellow solid.

LC-MS:m/z=210.0[M+H]⁺

Fourth step 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 well. Cooled to-5 ℃, sodium nitrite solution (497 mg,7.20mmol, dissolved in 4mL water) was slowly added dropwise thereto, and the reaction was continued at that temperature for 1 hour, and the solution was ready for use. Anhydrous copper dichloride (192 mg,1.42 mmol) was added to acetic acid (15 mL) and sulfur dioxide gas was continuously vented for 1 hour and cooled to 5 ℃. The diazonium salt solution was added to the reaction and reacted at-5-0 ℃ for 2 hours, and TLC detection (PE/ea=4/1, r _f =0.3) was complete. The reaction was quenched by addition to ice water (100 mL), extracted with dichloromethane (50 mL x 3), the organic phases combined, washed with water (100 mL x 2), dried over anhydrous sodium sulfate and concentrated to afford the title compound 21e (1.20, crude) as a brown oil, which was used directly in the next step.

Fifth step 3-bromo-4, 5-difluorobenzenesulfonamide 21f

25% -28% Ammonia (10 mL) was cooled to-5 ℃, a tetrahydrofuran solution of compound 21e (1.20 g crude product, dissolved in 5mL tetrahydrofuran) was added dropwise thereto, and after the addition, the reaction solution was slowly warmed to room temperature and reacted for 30 minutes, and TLC detection (PE/ea=4/1, r _f =0.8) was complete. The reaction was quenched by addition of saturated brine (50 mL), extracted with ethyl acetate (30 mL x 3), the combined organic phases dried over anhydrous sodium sulfate, concentrated, and the crude product purified by silica gel column chromatography (DCM/meoh=40/1) to give the title compound 21f as a yellow solid (735 mg, two step yield 55.6%).

¹H NMR(400MHz,DMSO-d₆)δ7.95(dt,J=5.6,2.0Hz,1H),7.09-7.85(m,1H),7.66(s,2H).

Sixth step 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), oxalyl chloride (1 mL,11.8 mmol) and DMF (1 drop) were added sequentially at room temperature, the reaction was completed at room temperature for 4 hours, the starting materials were detected by TLC (DCM/MeOH=20/1, R _f =0.2), the reaction solution was concentrated to dryness, 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 sequentially at room temperature, and the reaction was completed at room temperature for 2 hours. TLC detection (DCM/meoh=20/1, r _f =0.5) starting material reacted completely. The reaction was quenched with water (15 mL), extracted with dichloromethane (15 mL x 3), the combined organic phases dried over anhydrous sodium sulfate, concentrated, and the crude product purified by silica gel column chromatography (DCM/meoh=30/1) to give 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[M-H]^-(99.91%purity,210nm)

¹H NMR(400MHz,DMSO-d₆)δ8.15-8.02(m,2H),7.74(d,J＝2.0Hz,1H),7.61(d,J＝8.0Hz,1H),7.52(dd,J＝8.4,2.0Hz,1H).( Sulfonamide active hydrogens not shown).

Example 22

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

First step morpholine-4-sulfonamide 22b

Morpholine 22a (800 mg,9.18 mmol) was added to 1, 4-dioxane (6 mL), sulfonamide (1.06 g,11.0 mmol) was added thereto, the reaction mixture was heated to 120 ℃ and reacted for 18 hours after the addition, TLC monitoring (DCM/meoh=20/1, r _f =0.2) was complete and the reaction mixture was cooled to room temperature. The reaction solution was concentrated to dryness, to which was added ethyl acetate (20 mL), and the reaction solution was heated under reflux until complete dissolution. Filtering while the mixture is hot, and collecting filtrate. The filtrate was naturally cooled to room temperature, a large amount of solid was precipitated, filtered, rinsed with ethyl acetate (10 mL) and the filter cake was dried to give the title compound 22b (805 mg, yield 54.12%) as a yellow solid.

LC-MS:m/z=167.1[M+H]⁺

¹H NMR(400MHz,DMSO-d₆)δ6.82(s,2H),3.65(t,J=4.8Hz,4H),2.92(t,J=4.8Hz,4H)

Second step 2, 4-dichloro-N- (morpholinosulfonyl) benzamide 22

2, 4-Dichlorobenzoic acid (300 mg,1.57 mmol) was added to dichloromethane (10 mL), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (900 mg,2.36 mmol) and N, N-diisopropylethylamine (1 mL,5.42 mmol) were added thereto, the reaction was stirred at room temperature for 18 hours after the addition, TLC monitoring (DCM/MeOH=20/1, R _f =0.2) was complete, water (10 mL) was added thereto to extract the fractions, anhydrous sodium sulfate was dried, and the organic phase was concentrated to dryness. To this was added dichloromethane (10 mL) to dissolve, then compound 22b (240 mg,1.44 mmol) and N, N-diisopropylethylamine (1 mL,5.42 mmol) were added, and after the addition was completed, stirring was performed at room temperature for 18 hours, TLC monitoring (DCM/meoh=20/1, r _f =0.6) was complete, the reaction was quenched with water (10 mL), extracted with dichloromethane (10 mL x 2), and the organic phases were combined. Dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (DCM/meoh=80/1) to give 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)

¹H NMR(400MHz,DMSO-d₆)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).

Example 23

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

First step 3- (2-methoxyethyl) thiophene 23b

Sodium hydride (9.36 g, 60%) was added to anhydrous tetrahydrofuran (100 mL) under ice bath conditions, 3-thiopheneethanol 23a (10.0 g,78.0 mmol) was slowly added, methyl iodide (33.0 g,234 mmol) was slowly added at 0 ℃ after the addition, the reaction liquid nitrogen gas was allowed to react under protection for 2 hours at 60 ℃, TLC monitored the starting material (ethyl acetate/petroleum ether=1/10, r _f =0.2) was reacted completely, and new spots (ethyl acetate/petroleum ether=1/10, r _f =0.6) were formed. The reaction solution was cooled to room temperature, slowly poured into a saturated aqueous ammonium chloride solution (150 mL) at 0 ℃ and extracted with ethyl acetate (60 ml×3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 23b (13.6 g, crude) as a pale yellow liquid, which was directly subjected to the next step.

Second step 3- (2-methoxyethyl) thiophene-2-sulfonyl chloride 23c

The dry ice/ethanol bath was lowered to-10 ℃, and a dichloromethane solution (50 mL) of compound 23b (11.0 g, crude) was slowly added dropwise to a dichloromethane solution (50 mL) of chlorosulfonic acid (10 mL) (with vigorous warming, temperature control below 0 ℃). After the completion of the dropping, TLC monitoring the disappearance of the starting material (ethyl acetate/petroleum ether=1/10, r _f =0.6) and the formation of new points (ethyl acetate/petroleum ether=1/10, r _f =0.3). The reaction was slowly poured into ice water (80 mL) to quench, 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 give the title compound 23c (4.46 g, crude) as a pale yellow liquid, which was directly subjected to the next step.

Third step 3- (2-methoxyethyl) thiophene-2-sulfonamide 23d

A solution of compound 23C (4.46 g, crude) in tetrahydrofuran (20 mL) was slowly added dropwise to aqueous ammonia (20 mL) and reacted at 20 ℃ for 30 minutes, TLC monitored the disappearance of starting material (ethyl acetate/petroleum ether=1/10, r _f =0.3). The reaction solution was concentrated, the crude product was diluted with water (50 mL), ethyl acetate (20 mL x 2) was extracted, the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate/petroleum ether=1/3) to give the title compound 23d (200 mg, three-step yield 1%)

¹H NMR(400MHz,CDCl₃)δ7.45(d,J＝4.8Hz,1H),6.97(d,J＝5.2Hz,1H),5.39(s,2H),3.66(t,J＝5.6Hz,2H),3.32(s,3H),3.27(t,J＝5.6Hz,2H).

Fourth step 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 ℃ and reacted for 12 hours at 20 ℃ after addition, TLC monitored the disappearance of starting material (ethyl acetate/petroleum ether=1/3, r _f =0.3). The reaction mixture was quenched with water (5 mL), extracted with ethyl acetate (2 ml×2), the organic phases combined, washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product purified by Pre-TLC (ethyl acetate/petroleum ether=1/3) to give the title compound 23e (40 mg, yield 15%) as a pale yellow solid

LCMS:m/z=299.9[M+H]⁺

¹H NMR(400MHz,CDCl₃)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).

Fifth step 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), EDCI (33 mg,0.173 mmol) and HOBT (24 mg,0.173 mmol) were added sequentially at 20℃and the reaction was stirred for a further 1 hour, and TLC monitored for disappearance of starting material (dichloromethane/methanol=1/15, R _f =0.2). The reaction mixture was quenched with water (2 mL), extracted with dichloromethane (1 mL. Times.2), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was added compound 23e (40 mg,0.133 mmol), N-diisopropylethylamine (35 mg,0.266 mmol) and dichloromethane (0.5 mL), reacted at 20℃for 10 hours after the addition, TLC monitored compound 24e (ethyl acetate/petroleum ether=1/3, R _f =0.3) disappeared, the reaction solution was quenched with water (2 mL), extracted with dichloromethane (1 mL x 2), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. Purification of the crude product by Pre-TLC (dichloromethane/methanol=15/1) gave the title compound 23 (4 mg, 6% yield) as a white solid

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

¹H NMR(400MHz,DMSO-d₆)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).

Example 24

5- (2-Acetamido) -N- ((5-bromothiophen-2-yl) sulfonyl) -2, 4-dichlorobenzamide 24

First step 5- ((tert-Butoxycarbonyl) amino) -2, 4-dichlorobenzoic acid methyl ester 24b

Methyl 2, 4-dichloro-5-bromobenzoate 24a (12.3 g,43.3 mmol) was dissolved in toluene (120 mL), palladium acetate (4816 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 this order, the addition was completed, nitrogen was replaced three times, the reaction mixture was warmed to 120 ℃ and reacted for 20 hours, TLC monitored for disappearance of starting material (ethyl acetate/petroleum ether=1/5, r _f =0.5) and a new point was formed (ethyl acetate/petroleum ether=1/5, r _f =0.4). The reaction solution was cooled to room temperature, filtered, the filter cake was washed with ethyl acetate (30 mL x 2), the organic phases were combined, diluted with water (100 mL), extracted with ethyl acetate (50 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 give the title compound 24b (6.65 g, yield 47%)

Second step 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 ℃, the reaction was incubated for 1 hour, TLC monitored for disappearance of starting material (ethyl acetate/petroleum ether=1/5, r _f =0.4) and a new spot was formed (ethyl acetate/petroleum ether=1/5, r _f =0.2). The reaction solution was concentrated to about 30mL, diluted with water (50 mL), basified to ph=8-9 by dropwise addition of 30% sodium hydroxide solution at 0 ℃, extracted with ethyl acetate (30 ml×3), combined with the organic phase, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the title compound 24C (4.77 g, crude product) as a white solid, which was directly subjected to the next step.

Third step 5- (2-acetamido) -2, 4-dichlorobenzoic acid methyl ester 24d

Compound 24C (4.67 g, crude product) was dissolved in dichloromethane (45 mL), 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 this order at 20 ℃ and the reaction solution was reacted at 20 ℃ for 10 hours after the addition, and TLC monitored the disappearance of starting material (ethyl acetate/petroleum ether=1/1, r _f =0.6) and a new spot was formed (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 give the title compound 24d (11.2 g, crude product) as a pale yellow solid, which was used directly in the next step.

LCMS:m/z=319.0[M+H]⁺

Fourth step 5- (2-acetamido) -2, 4-dichlorobenzoic acid 24e

Compound 24d (10.7 g, crude) was dissolved in anhydrous methanol (100 mL), sodium hydroxide solid (1.34 g,33.5 mmol) and water (100 mL) were added at 10 ℃, and the reaction was continued for 3 hours at 20 ℃ after the addition, TLC monitored the disappearance of starting material (dichloromethane/methanol=10/1, r _f =0.7) with a new point formation (dichloromethane/methanol=5/1, r _f =0.1). The reaction was concentrated, acidified to ph=2-3 with hydrochloric acid (25 mL,6 n) at 0 ℃, extracted with ethyl acetate (40 ml×4), the organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, slurried at room temperature after concentration (ethyl acetate/petroleum ether=1/3, 50 mL), filtered, and the filter cake dried to give the title compound 24e as a pale yellow solid (7.46 g, three-step yield 73%).

Fifth step 5- (2-acetamido) -N- ((5-bromothiophen-2-yl) sulfonyl) -2, 4-dichlorobenzamide 24

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

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

¹H NMR(400MHz,DMSO-d₆)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).

Example 25

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

First step 5- (2-acetamido) -N- ((5-bromothiophen-2-yl) sulfonyl) -2, 4-dichlorobenzamide 25

Compound 24e (97 mg,0.32 mmol) was added to dichloromethane (1 mL), EDCI (80 mg,0.41 mmol) and HOBT (56 mg,0.0.41 mmol) were added sequentially at 20 ℃, and after addition, the reaction was stirred for further 1 hour and TLC monitored for disappearance of starting material (dichloromethane/methanol=1/15, r _f =0.2). The reaction mixture was quenched with water (2 mL), extracted with dichloromethane (1 mL. Times.2), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was added compound 23e (100 mg,0.33 mmol), N-diisopropylethylamine (205 mg,1.58 mmol) and dichloromethane (1 mL), and the reaction was continued for 10 hours at 20 ℃ after addition, TLC monitored compound 24e (ethyl acetate/petroleum ether=1/3, r _f =0.3) disappeared, the reaction was quenched with water (2 mL), extracted with dichloromethane (1 ml×2), the organic phases were combined, washed with saturated brine (1 mL), dried over anhydrous sodium sulfate, and concentrated. Purification of the crude product by Pre-TLC (dichloromethane/methanol=1/10) gave the title compound 25 (20 mg, 11% yield) as a white solid

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

¹H NMR(400MHz,DMSO-d₆)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,1H),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).

Example 26

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

First step 2- (benzylthio) -5-bromothiazole 26b

Benzyl mercaptan (6.14 g,49.40 mmol) was added to a solution of 2, 5-dibromothiazole 26a (10.0 g,41.17 mmol) in DMF (50 mL) of potassium carbonate (11.38 g,82.33 mmol), stirred at 25℃for 16 hours, TLC showed the end of the reaction (PE/EA=15/1, starting material R _f =0.5, product R _f =0.55). The reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (50 mL), washed with water (50 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude title compound 26b (10.6 g, 90.0% yield) as a pale yellow liquid by column chromatography over silica gel.

Second step 5-bromothiazole-2-sulfonyl chloride 26c

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) at 0℃and the reaction mixture was warmed to 25℃and stirred for 2 hours, TLC showed the end of the reaction (PE/EA=10/1, starting 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. Times.2), washed with saturated sodium bicarbonate solution (150 mL. Times.2), washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude title compound 26c (8.75 g, crude) as a pale yellow liquid, which was directly used in the next reaction.

Third step 5-bromothiazole-2-sulfonyl chloride 26d

Ammonia (5 mL) was added to a solution of compound 26C (8.75 g, crude) in tetrahydrofuran (50 mL) at 0 ℃, the reaction was warmed to 25 ℃ and stirred for 30min, TLC showed the reaction to end (PE/ea=3/1, starting material R _f =0.5, product R _f =0.3). The reaction was concentrated, acidified to ph=3-4 with 2M hydrochloric acid, extracted with ethyl acetate (50 mL), washed with water (5 mL x 2), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude title compound 26d (2.76 g, 32.2% yield in two steps) as a white solid by separation and purification on a silica gel column.

LC-MS:m/z=242.9[M+H]⁺

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

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) at 0℃and the reaction was warmed to 25℃and reacted for 1 hour, the reaction concentrated and the concentrate dissolved in DCM (2 mL). A solution of the concentrate in DCM was added to compound 26d (150 mg,0.62 mmol) and triethylamine (87 mg,1.85 mmol) in DCM (3 mL) at 0deg.C, warmed to 25deg.C and stirred for 30 min, TLC showed complete reaction (PE/EA=2/1, starting material R _f =0.3, DCM/MeOH=10:1, product R _f =0.1), the reaction concentrated, the crude was dissolved in water (15 mL), extracted with PE/EA (3/1, 15mL x 3), aqueous phase 2M hydrochloric acid acidified to pH=3-4, extracted with ethyl acetate (15 mL), water (5 mL x 2) dried, filtered, the filtrate concentrated, the crude was suspended in DCM (5 mL), and PE (15 mL) was added and stirred for 15min, filtered, the filter cake was washed with PE/EA (3/1, 2 mL) and dried to give 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)

¹H NMR(400MHz,DMSO-d₆)δ8.24(s,1H),7.72(d,J＝1.6Hz,1H),7.59(d,J＝8.0Hz,1H),7.51(dd,J＝8.4,2.0Hz,1H).

Example 27

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

First step 5- ((trimethylsilyl) ethynyl) thiazole-2-sulfonamide 27a

Cuprous iodide (20 mg,0.1 mmol) and Pd (PPh ₃)₂Cl₂ (72 mg,0.1 mol) were added to a solution of compound 26d (500 mg,2.06 mmol) and triethylamine (264 mg,6.17 mmol) in DMF (5 mL), the protection was replaced with nitrogen three times, trimethylethynyl silicon (1.14 g,11.57 mmol) was added to the reaction solution by syringe, the reaction solution was warmed to 80 ℃ and stirred for 30 min, TLC showed the end of the reaction (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 was precipitated, filtered, the filter cake was washed with EA (5 mL), the filtrate was washed with water (50 ml×3), the EA phase was dried with anhydrous sodium sulfate, filtered, concentrated to give the crude product, which was separated by silica gel column chromatography to purify the title compound 27a (310 mg, 57.9%) as pale yellow liquid.

LC-MS:m/z=261.0[M+H]⁺

Second step 5-ethynyl thiazole-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 25deg.C, the reaction stirred for 30min, and TLC showed the end of the reaction (PE/EA=5/1, starting material R _f =0.5, product R _f =0.3). The reaction 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×2), the organic phase dried over anhydrous sodium sulfate, filtered and the filtrate concentrated to give the title compound 27b (108 mg, 100%) as a pale yellow liquid.

LC-MS:m/z=189.0[M+H]⁺

Third step 2, 4-dichloro-N- ((5-ethynyl thiazol-2-yl) sulfonyl) benzamide 27

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) at 0℃and the reaction was warmed to 25℃and reacted for 1 hour, the reaction concentrated and the concentrate dissolved in DCM (2 mL). The above solution was added to compound 27b (108 mg,0.57 mmol) and triethylamine (174 mg,1.72 mmol) in DCM (3 mL) at 0 ℃, the reaction was warmed to 25 ℃ and stirred for 30 min, TLC showed complete reaction (PE/ea=2/1, starting material R _f =0.3, DCM/meoh=10:1, product R _f =0.1), the reaction concentrated, the crude was dissolved in water (15 mL), the PE/EA mixture (3/1, 15mL x 3) extracted, aqueous phase 2M hydrochloric acid acidified to ph=3-4, ethyl acetate (15 mL) extracted, water (5 mL x 2) washed, the organic phase dried over anhydrous sodium sulfate, filtered, the filtrate concentrated, the crude was suspended in DCM (5 mL), petroleum ether (15 mL) was added and stirred for 15 min, filtered, the filter cake was washed with PE/EA (3/1, 2 mL) to afford 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)

¹H NMR(400MHz,DMSO-d₆)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 at molecular level with WDR5

Affinity testing of compounds of the present disclosure with WDR5 at the molecular level was performed by the following method:

Compound preparation compound was accurately weighed and dissolved to 10mM stock solution using DMSO (Sigma, D5879). The mother liquor was diluted 2-fold in a gradient to 9 concentrations using 1 XHBS-EP buffer (10mM HEPES,pH 7.4,150mM NaCl,3.0mM EDTA,and 0.005% (v/v) TW-20), the final concentration of the compounds in the reaction system was 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78, 0.39. Mu.M, and the final concentration of the DMSO of the compounds was 5%. A number of compound samples were sequentially transferred to 96-well plates (Greiner, 650101) according to the corresponding experimental program design.

Surface plasmon resonance experiments (SPR) WDR5 proteins were coupled to CM5 chips (Cytiva, BR 100530) using an amino coupling method with a signal value of about 9000 Response Units (RU). The three initialization cycles were first run, followed by sequential injection of the compounds to be tested, setting a binding time of 120 seconds and a dissociation time of 100 seconds, and then washing out the residual compounds in the chip using a solution containing 50% DMSO. Solvent correction was performed every 48 cycles using 1x HBS-EP buffer with 4.5% -5.8% dmso.

Detection and analysis signal detection, data collection and processing analysis were performed using Biacore 8K. The raw data generated by the experiment were corrected by subtracting the control group parameters and solvent using Biacore8K data processing software, and the corresponding K _D values were fitted using a static affinity model, as detailed in table 1.

Conclusion the compounds of the examples of the present disclosure have a stronger affinity at the molecular level for WDR5 proteins.

TABLE 1 affinity test results of compounds of the present disclosure with WDR5 protein

Numbering of compounds	Affinity (K _D, mu M)	Numbering of compounds	Affinity (K _D, mu M)
				1	18.7	11	9.8
2	10.4	12	5.4
				3	65.5	13	5.3
4	21.0	14	2.5
				5	31.2	15	24.1
6	36.4	16	7.2
				7	15.6	17	20.8
8	5.3	20	129.0
				9	2.42	21	23.1

Test example 2 anti-proliferation Activity of Compounds against neuroblastoma cells

The anti-proliferation activity test of the compounds in the present disclosure on neuroblastoma cells was performed by the following method:

Compound preparation compound was accurately weighed and dissolved to 10mM stock solution using DMSO for use. Compounds were diluted to a final test concentration of 10. Mu.M using MEM or RPMI 1640 medium and DMSO to a final concentration of 0.1% and transferred to 96 well plates for subsequent experiments.

Cell counting kit-8 test (CCK-8) the 293T, SK-N-AS, IMR32 and LAN5 cells were seeded into 96 well cell culture plates at 2X10 ⁴ per well and incubated for 72 hours after addition of the corresponding compounds and DMSO controls per well. After that, 10. Mu.L of CCK-8 reagent was added to each well, and incubation at 37℃was continued for 2 hours. The absorbance of the cells at 450nm was measured using a microplate detector.

Detection and analysis, collecting absorbance data corresponding to each well of cells, and calculating corresponding cell survival rate and inhibition rate, as detailed in Table 2.

TABLE 2 results of antiproliferative activity of certain compounds of the present disclosure on neuroblastoma cells

Conclusion the compounds of the examples of the present disclosure have good antiproliferative activity against neuroblastoma cells. Wherein "-" indicates no activity tested.

Test example 3 Complex structural resolution of a portion of the Compounds of the present disclosure with WDR5

The resolution of the complex structure of selected compounds 13 and 14 in the present disclosure is performed by the following method:

Protein crystal preparation, namely placing WDR5 expression vector Rosetta2 competent cells in a 37 ℃ constant temperature shaking incubator to culture until the OD value is 0.6-0.8, adding IPTG with the final concentration of 0.1mM, and continuously culturing at 16 ℃ for 20 hours to collect the cells. Cells were disrupted using a high pressure cell disruptor, and the supernatant was removed after centrifugation at high speed for 30 minutes. 10mL of Ni-NTA affinity chromatography gel resin is added, stirred and combined for 1 hour at 4 ℃, and the supernatant is centrifugally taken and treated with a purifying buffer (50mM Tris pH 7.5,150mM NaCl,50mM imidazole, 1mM TCEP) and an eluting buffer (50mM Tris pH 7.5,150mM NaCl,300mM imidazole, 1mM TCEP) respectively to obtain the preliminary target protein. And then respectively using an anion exchange chromatographic column and a size exclusion chromatographic column to obtain high-purity WDR5 protein, collecting a sample, and using SDS-polyacrylamide gel electrophoresis to identify the purity of the protein.

The obtained high purity WDR5 protein was concentrated to 11mg/mL using an ultrafiltration tube and centrifuged at 12000rpm for 30 minutes. A crystallization buffer, 0.1M Bis-Tris, pH 5.8,0.2M ammonium acetate, 30% (w/v) pEG3350 was prepared. Crystallization condition optimization was performed using a 24-well hanging-drop plate. 1mL of buffer per well. The two solutions were mixed in a ratio of 1.5. Mu.L of crystallization buffer+1.5. Mu.L of protein solution, and crystals were grown in a plate-like form after 2-3 days in an incubator at 18 ℃. The composite crystal of WDR5 and active compound is obtained by soaking method.

The diffraction data of the crystals were collected at the open sea synchrotron radiation light source (SSRF) microcrystal compound line station BL18U 1. The collected data is processed by HKL3000, which comprises the steps of data point selection, indexing, correction, integration, data merging, normalization and the like, and an MTZ file is generated. The WDR5 empty protein crystal structure analyzed in the laboratory is used as a template, and the Phaser module in CCP4 is used for carrying out molecular replacement to generate initial structure coordinates. Model construction and optimization are carried out by using COOT, and then further optimization is carried out by using Phenix, and R factor in structural parameters is made to be smaller than 0.25 repeatedly. The results are shown in FIG. 1.

Conclusion compounds of the examples of the present disclosure bind to WDR5 by occupying the WBM site to block WDR5 interaction with Myc.

Test example 4 in vitro potency test of some of the compounds of the present disclosure against acute myelogenous leukemia, hepatoma cell lines and pancreatic cancer cell lines

Cell counting kit-8 test (CCK-8) Hep3B, huh-7, MV4-11 and Mia-Paca2 cells were seeded into 96-well cell culture plates at 2X10 ⁴ cells/well, and incubated for 72 hours after addition of the corresponding compounds and DMSO controls per well. After that, 10. Mu.L of CCK-8 reagent was added to each well, and incubation at 37℃was continued for 2 hours. The absorbance of the cells at 450nm was measured using a microplate detector.

Detection and analysis, collecting absorbance data corresponding to each well of cells, and calculating corresponding inhibition rate, as shown in Table 3.

TABLE 3 results of in vitro potency test of some of the compounds of the present disclosure on acute myelogenous leukemia, hepatoma cell lines and pancreatic cancer cell lines

Conclusion the compounds of the examples of the present disclosure have good antiproliferative activity against hepatoma cells, acute myeloid leukemia and pancreatic cancer cells.

Test example 5 pharmaceutical substitution test of some of the compounds of the present disclosure on Normal mice

The pharmacokinetic properties of Compound 11 on mice were studied in this test example, using Balb/c female mice, 6-8 weeks old, and each of which was administered by gastric lavage or tail vein injection of Compound 11. Whole blood was collected 5min, 15min, 30min, 1h, 2h, 4h, 8h, 24h after dosing, respectively, and after centrifugation at 8000rpm for 5min, the upper plasma was collected, and the drug was tested for plasma exposure at different time points by HPLC-MS/MS method, and pharmacokinetic parameters were calculated. The specific experimental results are as follows:

TABLE 4 results of drug substitution test of some of the compounds of the present disclosure on normal mice

Conclusion the compound of the embodiment of the disclosure has rapid oral absorption and extremely high bioavailability, and the half-life T _1/2 of plasma is as long as 15.9h, thus being an ideal potential oral drug candidate molecule.

Test example 6 efficacy test of some compounds of the present disclosure against IMR32 CDX mouse neuroblastoma model

The drug effect of the compound 11 on a neuroblastoma model is studied, and human neuroblastoma IMR32 cells are selected for in vitro monolayer culture, and conventional digestion treatment and passage are carried out twice a week by using pancreatin-EDTA. When the saturation of the cells is 80% -90%, collecting the cells, counting, adjusting the inoculation concentration to 10 million/animal, inoculating the cells under the back skin of the upper right side of the animal, and carrying out random grouping and administration according to the weight and the tumor volume of the animal when the tumor grows to about 120mm ³. Animal body weight and tumor volume were measured twice weekly. The efficacy data are expressed as Tumor Volume (TV), tumor inhibition rate (TGI), relative volume (T/C), and the specific experimental data are as follows:

table 5. Results of potency test of partial compounds of the present disclosure on IMR32 CDX mouse neuroblastoma model n=6

Experimental results show that the animal health state is not obviously abnormal during the administration period, and compared with a solvent control (Vehicle) group, the compound of the embodiment of the disclosure has obvious anti-tumor drug effect in three dosage groups of low, medium and high, and has good dose-effect relationship, the TGI reaches 85.5%, 107.1% and 107.1% respectively, and part of animals in the high and medium dosage groups can completely disappear in the later period of administration. It can be seen that the compound of the embodiment of the disclosure has remarkable effects in an IMR32 human neuroblastoma model.

Test example 7 test of efficacy of some compounds of the present disclosure against Hep3B CDX mouse hepatoma model

The test example is used for researching the drug effect of the compound 11 on a liver cancer model, and selecting human liver cancer Hep3B cells, carrying out in vitro monolayer culture, and carrying out conventional digestion treatment passage by using pancreatin-EDTA twice a week. When the saturation of the cells is 80% -90%, collecting the cells, counting, adjusting the inoculation concentration to 10million cells/animal, inoculating the cells to the back of the upper right side of the animal to be measured subcutaneously, and carrying out random grouping and administration according to the weight and the tumor volume of the animal when the tumor grows to about 110mm ³. Animal body weight and tumor volume were measured twice weekly. The efficacy data are expressed as Tumor Volume (TV), tumor inhibition rate (TGI), relative volume (T/C), and the specific experimental data are as follows:

Table 6. Results of test of efficacy of partial compounds of the present disclosure on Hep3B CDX mouse hepatoma model n=5

The experimental result shows that the animal health state is not obviously abnormal during the administration period, and compared with a solvent control (Vehicle) group, the compound in the embodiment of the disclosure has obvious anti-tumor drug effect in both low and high dose groups and good dose-effect relationship, the TGI of the compound reaches 40% and 106.8% respectively, and the tumor of part of animals in the high dose 90mg/kg group can completely disappear in the later period of administration. It can be seen that the compound of the embodiment of the disclosure has remarkable effects in a Hep3B liver cancer xenograft tumor model.

Test example 8 safety test of some of the compounds of the present disclosure on normal mice

The safety of compound 11 was studied in this test example, and female, 6-8 week old ICR mice were selected as subjects, and the animals were randomized according to their body weights and then orally administered at 100mg/kg, 300mg/kg, 400mg/kg, 500mg/kg, respectively, to observe their body weight changes, morbidity and mortality, as follows:

TABLE 7 safety test results of some of the compounds of the present disclosure on normal mice

Group of	Number of animals	Dead mice	Death/total number	Time to death
					Compound 11,100mg/kg	3	0	0/3	NA
Compound 11,300mg/kg	3	0	0/3	NA
					Compound 11,400mg/kg	3	0	0/3	NA
Compound 11,500mg/kg	3	1	1/3	Day3

Experimental results show that on ICR mice, all animals do not cause morbidity or mortality at doses of 100mg/kg,300mg/kg and 400mg/kg of the compound of the embodiment of the disclosure, and only 1 in the 500mg/kg group dies on the 3 rd day of the experiment, so that the compound of the embodiment of the disclosure expects tolerance doses as high as 400mg/kg, and has good safety.

Test example 9 protein binding Rate test of some of the compounds of the present disclosure

The plasma protein binding rate of the compound 11 in human plasma is measured by adopting an equilibrium dialysis method, a semipermeable membrane is utilized to separate a left chamber from a right chamber, a protein solution containing medicine is added to the left side, a blank buffer solution is added to the right side, unbound free medicine can freely pass through the semipermeable membrane, after a certain period of incubation, the two sides reach equilibrium, the concentration of the free medicine is equal, and the plasma protein binding rate can be calculated by measuring the concentration of the medicine on the two sides. The results are shown in Table 8:

TABLE 8 protein binding Rate test results for some of the compounds of the present disclosure

Numbering of compounds	Species of genus	Binding Rate (%)	Free fraction (%)	Recovery (%)
					11	Human body	98.52	1.48	88.33

Experimental results show that the free drug of the compound in the embodiment of the disclosure reaches 1.48%, and the compound is enough to meet the in-vivo therapeutic drug concentration requirement.

Test example 10 hepatocyte stability test of some of the compounds of the present disclosure

Transfer 198 μl of viable cell suspension to 96-well deep well plates and place the deep well plates on vortex for 10min pre-heating in incubator. The experiments were performed with double parallel incubations. 2. Mu.L of 100. Mu.M of the test substance or verapamil positive drug was added to each well to initiate the reaction, and the deep well plate was returned to the incubator shaker. Samples were incubated for 0, 15, 30, 60, 90 and 120 minutes, 25. Mu.L of the suspension was taken and the reaction was stopped by the addition of 150. Mu.L of acetonitrile containing the internal standard. Vortex for 10min and centrifuge at 3220g, 4 ℃ for 45 min. Transfer 100 μl of supernatant to sample plate, add 100 μl of purified water and mix well for UPLC-MS/MS analysis. The results are shown in Table 9:

TABLE 9 hepatocyte stability test results for some of the compounds of the present disclosure

Experimental results show that the compound of the embodiment of the disclosure is easy to be metabolically cleared in human liver and is not easy to generate accumulated toxicity.

Applicant states that the disclosure illustrates, by way of the above examples, a sulfonamide compound of the disclosure as an interaction blocker of WDR5 and Myc, and methods of making and using the same, but the disclosure is not limited to, i.e., does not mean that the disclosure must rely on, to be practiced. It should be apparent to those skilled in the art that any modifications of the present disclosure, equivalent substitutions of raw materials for the products of the present disclosure, addition of auxiliary components, selection of specific modes, etc., are within the scope of the present disclosure and the scope of the disclosure. The above describes in detail the optional embodiments of the present disclosure, but the present disclosure is not limited to the specific details in the above embodiments, and various simple modifications may be made to the technical solutions of the present disclosure within the scope of the technical concept of the present disclosure, and all the simple modifications belong to the protection scope of the present disclosure. In addition, the specific features described in the foregoing embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, the present disclosure does not further describe various possible combinations.

Claims

1. 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.

2. 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.

3. 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.

4. The sulfonamide compound according to claim 1 or a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, wherein the sulfonamide compound represented by formula (I) is the following compound:

5. A pharmaceutical composition, comprising a sulfonamide compound as described in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, 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.

6. Use of the sulfonamide compound according to any one of claims 1 to 4 or its pharmaceutically acceptable salt, its tautomer or its stereoisomer 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.

7. 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.

8. 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.