CN111285806A - Pyrazole compound and preparation method, application and pharmaceutical composition thereof - Google Patents

Pyrazole compound and preparation method, application and pharmaceutical composition thereof Download PDF

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CN111285806A
CN111285806A CN201811484464.9A CN201811484464A CN111285806A CN 111285806 A CN111285806 A CN 111285806A CN 201811484464 A CN201811484464 A CN 201811484464A CN 111285806 A CN111285806 A CN 111285806A
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许恒
陈晓光
林松文
季鸣
张敬博
杜婷婷
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Abstract

The invention relates to a substituted pyrazole compound, a preparation method, application and a pharmaceutical composition thereof. The pyrazole compound is shown as a formula (I), is an STAT3 signal pathway inhibitor, and can be used for preventing and/or treating diseases related to STAT3 activity, such as tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammatory diseases, metabolism/endocrine dysfunction or neurological diseases.

Description

Pyrazole compound and preparation method, application and pharmaceutical composition thereof
Technical Field
The invention belongs to the technical field of pharmacy, and relates to a pyrazole compound, and a preparation method, application and a pharmaceutical composition thereof.
Background
The Signal Transducer and Activator of Transcription (STAT) protein family performs this dual function of Signal transduction and transcriptional regulation. Although the members of the STAT family are structurally similar, they are involved in different cellular processes. There are 7 STAT family members that have been isolated and purified so far, namely STAT1, STAT2, STAT3, STAT4, STAT5A, STAT5B and STAT 6.
As a member of the STAT family, STAT3 plays an important role in the development and progression of cancer, inflammation, ischemia/reperfusion injury, and stem cell self-renewal. STAT3 can be activated by receptor tyrosine kinases and non-receptor tyrosine kinases. When the cytokine is combined with the receptor, the receptor is dimerized to cause phosphorylation of JAK kinase, so that tyrosine residue (Tyr705) at the C-terminal of STAT3 molecule is phosphorylated, and is activated by forming a dimer through SH2 region, and the receptor is transferred into nucleus to be combined with a specific DNA sequence to regulate and control the transcription of target genes. In addition, Ser727, located in the Stat3 transcriptional activation domain, is further activated by the MAPK or mTOR pathway, regulating Stat3 transcriptional activity, which is thought to be necessary for its full activation. In addition to phosphorylation of Tyr705 and Ser727, acetylation of Lys685 stabilizes Stat3 dimer, modulating Stat3 activity.
Currently, several STAT3 inhibitors have entered clinical research for tumor therapy and autoimmune diseases. STAT3 inhibitor napabucain, developed by boston biomedical corporation, qualifies as an FDA orphan drug for the treatment of gastroesophageal junction cancer at 2016, 6 months, currently in its clinical trial stage III; FDA orphan drug qualification for the treatment of pancreatic cancer is currently in its clinical trial stage III at 11 months of 2016. Still other STAT3 inhibitors are also being studied in various clinical trials, such as the STAT3 inhibitor GLG-302 developed by GLG company in phase I clinical trials for the treatment of tumors, polycystic kidney disease; GLG-801 is used in the treatment of renal leukemia, psoriasis, in phase II clinical trials. The STAT3 inhibitor MOL-4249 developed by the Moleculin biotechnology is used for treating mild-to-moderate psoriasis and is currently in the phase II clinical trial stage; AK-114 from Takeda is used for the treatment of ulcerative colitis and is in phase II clinical studies. In addition, Bt354 is reported in the literature to be used as an inhibitor of STAT3 signaling pathway and can be used for treating cancers such as triple negative breast cancer.
STAT3 has become a very attractive drug target, but there is also a need to develop safer and more effective STAT3 inhibitors for the prevention and/or treatment of tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammatory diseases, metabolic/endocrine dysfunction or neurological diseases.
Disclosure of Invention
The technical problem solved by the invention is to provide a novel STAT3 signal pathway inhibitor, a preparation method, a pharmaceutical composition and an application thereof, wherein the STAT3 signal pathway inhibitor has stronger inhibition activity on tumor cells with high STAT expression, particularly human prostate cancer cell DU-145, so that the STAT3 signal pathway inhibitor has better prevention and/or treatment effects on diseases mediated by STAT3, such as tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammatory diseases, metabolism/endocrine dysfunction or neurological diseases.
In order to solve the technical problem, the invention provides the following technical scheme:
in a first aspect of the technical scheme of the present invention, there is provided a compound represented by formula (I), a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof:
Figure BDA0001894123760000021
wherein
R1Is selected from C1-4Alkyl radical, C3-6A cycloalkyl group;
R2selected from: phenyl, wherein the phenyl is substituted at the para-position with Ra; 5-10 membered heteroaryl, wherein the 5-10 membered heteroaryl is substituted with Ra; or with C4-6Cyclo-olefins or C4-6Heterocyclic olefin fused phenyl;
ra is selected from: chlorine, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy and ethoxy;
R3and R4Each independently selected from hydrogen, halogen, methyl, ethyl and methoxy.
In a further preferred embodiment, R1Selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl.
In another preferred embodiment, the present invention provides a compound of formula (I), a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof:
wherein
R1Is selected from C1-4Alkyl radical, C3-6A cycloalkyl group; preferably selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl;
R2selected from:
Figure BDA0001894123760000022
ra is selected from the group consisting of: chlorine, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy and ethoxy;
R3and R4Each independentlySelected from hydrogen, halogen, methyl, ethyl and methoxy.
In a further preferred embodiment, Ra is selected from the group consisting of: methyl, ethyl, cyclopropyl, methoxy and ethoxy.
In yet another preferred embodiment, the present invention provides a compound of formula (I), a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof:
wherein
R1Is selected from C1-4Alkyl radical, C3-6A cycloalkyl group; preferably selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl;
R2selected from:
Figure BDA0001894123760000031
R3and R4Each independently selected from hydrogen, halogen, methyl, ethyl and methoxy.
In yet another preferred embodiment, the present invention provides a compound of formula (I), a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof:
wherein
R1Is selected from C1-4Alkyl radical, C3-6A cycloalkyl group; preferably selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl;
R2selected from phenyl, wherein said phenyl is substituted in the para position with Ra; 5-10 membered heteroaryl, wherein the 5-10 membered heteroaryl is substituted with Ra; or with C4-6Cyclo-olefins or C4-6Heterocyclic olefin fused phenyl;
ra is selected from: chlorine, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy and ethoxy;
preferably, R2Selected from:
Figure BDA0001894123760000032
ra is selected from the group consisting of: chlorine, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy and ethoxy;
further preferably, Ra is selected from the group consisting of: methyl, ethyl, cyclopropyl, methoxy and ethoxy;
or R2Selected from:
Figure BDA0001894123760000033
R3and R4Each independently selected from hydrogen, halogen, methyl, ethyl and methoxy.
In a further preferred embodiment, R3And R4Each independently selected from hydrogen, chlorine, fluorine, methyl, ethyl, and R3And R4At least one is hydrogen.
In a further preferred embodiment, R3Selected from methyl, R4Selected from hydrogen; or R3Selected from hydrogen, R4Selected from chlorine and fluorine.
In particular, preferred compounds according to the invention are the following:
Figure BDA0001894123760000041
in a second aspect of the present invention, there is provided a process for the preparation of said compounds, stereoisomers, geometric isomers, tautomers thereof, comprising the steps of:
Figure BDA0001894123760000051
(1) taking the compounds A and B as starting materials, and preparing a compound C through a condensation reaction;
(2) reacting compound C with arylsulfonyl chloride or heteroarylsulfonyl chloride to prepare a compound of formula (I), a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof.
The second aspect of the present technical solution also provides another method for preparing the compound, its stereoisomers, geometric isomers and tautomers, which comprises the following steps:
Figure BDA0001894123760000052
(1) taking the compound D as a starting material, and preparing a compound E through a sulfonylation reaction;
(2) hydrolyzing the compound E with lithium hydroxide to prepare a compound F;
(3) reacting compound F with (1-R)1-1H-pyrazol-4-yl) methylamine or a hydrochloride thereof, to prepare a compound of formula (I), a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof.
A third aspect of the present invention provides a pharmaceutical composition comprising said compound, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier and/or excipient; preferably, the pharmaceutical composition further comprises one or more pharmaceutically active ingredients for preventing and/or treating tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammatory diseases, metabolism/endocrine function disorders or neurological diseases, other than the compound, its stereoisomer, geometric isomer, tautomer or pharmaceutically acceptable salt thereof; preferably, the pharmaceutical composition is a pharmaceutically acceptable pharmaceutical preparation for preventing and/or treating tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammatory diseases, metabolism/endocrine dysfunction or neurological diseases.
In yet another aspect, the present invention also provides a pharmaceutical formulation comprising at least one of the compounds, stereoisomers, geometric isomers, tautomers or pharmaceutically acceptable salts thereof, and optionally a pharmaceutically acceptable carrier or excipient; preferably, the pharmaceutical formulation is selected from the following pharmaceutical dosage forms: parenteral formulations, such as injection solutions or suspensions; enterally, e.g., orally, such as tablets or capsules; formulations for topical administration, for example lotions, gels, ointments, creams, nasal preparations, suppositories, transdermal preparations or ophthalmic preparations.
In still another aspect, the present invention also provides a use of the compound, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition for the preparation of a medicament for the prevention and/or treatment of tumor, autoimmune disease, kidney disease, cardiovascular disease, inflammatory disease, metabolism/endocrine function disorder, or neurological disease. In other words, the present invention provides a method for preventing and/or treating tumors, autoimmune diseases, renal diseases, cardiovascular diseases, inflammatory diseases, metabolic/endocrine dysfunctions or neurological diseases, which comprises administering to a subject in need thereof a prophylactically and/or therapeutically effective amount of the compound, a stereoisomer, geometric isomer, tautomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition.
Some of the terms used in the present invention are defined below, and other undefined terms have meanings well known to those skilled in the art.
Halogen means fluorine, chlorine, bromine or iodine.
C1-4Alkyl refers to straight and branched chain saturated aliphatic hydrocarbon groups having 1 to 4 carbon atoms. Examples of such groups include, but are not limited to: methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.
C3-6Cycloalkyl refers to a saturated monocyclic, fused, spiro or polycyclic structure having 3 to 6 carbon ring atoms. Examples of such groups include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
C4-6Cyclic olefins refer to monocyclic structures having 4 to 6 carbon ring atoms containing one C ═ C double bond. Examples of such groups include, but are not limited to: cyclobutene, cyclopentene and cyclohexene.
C4-6A heterocycloalkene refers to a monocyclic structure containing one C ═ C double bond with 4 to 6 ring atoms, where one or more ring atoms are selectedFrom nitrogen, oxygen or S (O)m(wherein m is an integer of 0 to 2) but excludes the ring moiety of-O-O-, -O-S-or-S-S-, the remaining ring atoms being carbon. Examples of such groups include, but are not limited to: 2, 3-dihydrofuran, 2, 5-dihydrofuran, 1, 3-dioxole, 1, 4-dioxa-2-hexene, 2, 3-dihydro-1H-pyrrole, 2, 5-dihydro-1H-pyrrole, 1,2,3, 4-tetrahydropyridine and 1,2,3, 6-tetrahydropyridine.
Aryl refers to a monocyclic or bicyclic aromatic hydrocarbon group, typically having 6 to 10 carbon atoms. Examples of such groups include phenyl or naphthyl; phenyl is preferred.
Heteroaryl refers to 5-10 membered aromatic heterocyclic groups including, but not limited to: 5-membered heteroaryl: furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, triazolyl (1,2, 4-triazolyl, 1,3, 4-triazolyl or 1,2, 3-triazolyl), thiadiazolyl (1,3, 4-thiadiazolyl, 1,2, 5-thiadiazolyl, 1,2, 3-thiadiazolyl or 1,2, 4-thiadiazolyl), and oxadiazolyl (1,3, 4-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,2, 3-oxadiazolyl or 1,2, 4-oxadiazolyl); and 6-membered heteroaryl: pyridyl, pyrimidinyl, pyrazinyl and pyridazinyl; and bicyclic groups such as benzofuranyl, benzothienyl, indazolyl, purinyl, quinolyl, isoquinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, indolizinyl, indolyl, isoindolyl. Preferred heteroaryl groups are thienyl, thiazolyl, pyridyl, pyrimidinyl.
"optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, "alkyl optionally substituted with halogen" means that halogen may, but need not, be present, and the description includes the case where alkyl is substituted with halogen and the case where alkyl is not substituted with halogen.
The compounds of the invention may contain one or more chiral centers, which exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, and atropisomers, and mixtures thereof (e.g., racemic mixtures), are within the scope of the present invention.
The compounds of the present invention include geometric isomers thereof. For example, if the compounds of the present invention contain double bonds or fused rings, these compounds may exist as geometric isomers, and their cis, trans forms and mixtures of cis and trans forms are included in the scope of the present invention.
The compounds of the present invention include tautomers thereof. Tautomers refer to structural isomers of different energies that interconvert via a low energy barrier, such as keto-enol and imine-enamine tautomerism.
The compounds of the present invention also include isotopically-labeled compounds thereof, in which one or more atoms are replaced by an atom having the same atomic number, but an atomic mass number different from the atomic mass number usually found in nature. Examples of isotopes suitable for the invention include, but are not limited to: isotopes of hydrogen2H and3h; isotopes of carbon11C、13C and14c; isotopes of chlorine36Cl; isotopes of fluorine18F; isotope of iodine123I and125i; isotopes of nitrogen13N and15n; isotopes of oxygen15O、17O and18o; isotopes of phosphorus32Isotopes of P and sulfur35S。
Various hydrates and solvates of the compound or salt thereof of the present invention and polymorphic forms thereof (polymorphisms) are also included in the scope of the present invention.
Prodrugs of the compounds described herein are also included within the scope of the invention. Some derivatives of the compounds described in the present invention have weak pharmacological activity or no pharmacological activity themselves, but when these derivatives are administered into or onto the body, they may be converted into the compounds described in the present invention having pharmacological activity by means of, for example, hydrolytic cleavage, and the like, and these derivatives are referred to as "prodrugs". Further information on the use of prodrugs can be found in Pro-drugs as Novel delivery systems, vol.14, ACS Symposium Series (t.higuchi and w.stella) and BioreversibleCarriers in Drug Design, Pergamon Press,1987(ed.e.b.roche, american pharmaceutical Association).
The compounds of the present invention include pharmaceutically acceptable salts thereof. Pharmaceutically acceptable salts are salts that are pharmaceutically acceptable and possess the desired pharmacological activity of the parent compound. Pharmaceutically acceptable salts are described in detail in j.pharma.sci.,1977,66,1-19 by Berge et al, which is incorporated herein by reference. The compounds of the present invention may contain sufficient acidic groups, sufficient basic groups, or both types of functional groups, and accordingly react with some inorganic or organic bases, or inorganic and organic acids, to form pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, hydrochloride, hydrobromide, hydroiodide, acetate, propionate, caprate, caprylate, acrylate, formate, isobutyrate, hexanoate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1, 4-dioate, hexyne-1, 6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, dihydrogenphosphate, metaphosphate, hydrochloride, hydrobromide, hydroiodide, acetate, propionate, caprylate, or a mixture thereof, Citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate and mandelate.
When used as a medicament, the compounds of the present invention are generally administered in the form of a pharmaceutical composition. Accordingly, pharmaceutical compositions of the compounds of the present invention and a pharmaceutically acceptable carrier, diluent or excipient are also included within the scope of the present invention. Carriers, adjuvants, and excipients used herein include any and all solvents, diluents, or other liquid excipients, dispersing or suspending agents, surfactants, isotonic agents, thickening agents, as appropriate to the particular dosage form desiredAgents or emulsifiers, preservatives, solid binders, lubricants and the like. In Remington: the Science and Practice of Pharmacy, 21stedition，2005，ed.D.B.Troy，Lippincott Williams&Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds.J.Swarbrick and J.C.Boylan, 1988. 1999, Marcel Dekker, New York, disclose various carriers for the formulation of pharmaceutically acceptable compositions and known techniques for their preparation, the contents of which are incorporated herein by reference.
The compositions of the present invention may be administered by any route suitable for the condition to be treated. In particular by administration in the form: parenterally, e.g., in the form of injectable solutions or suspensions; enterally, e.g., orally, e.g., in tablet or capsule form; topically, for example in the form of a lotion, gel, ointment or cream or in the form of a nasal or suppository. Topical application is, for example, application to the skin. Another form of topical administration is to the eye.
The pharmaceutical compositions may be administered in solid, semi-solid, liquid or gaseous form, or may be in a dry powder, such as lyophilized form. Pharmaceutical compositions can be packaged in a form convenient for delivery, including, for example, solid dosage forms such as capsules, sachets, cachets, gelatin, paper, tablets, suppositories, pellets, pills, troches, and lozenges. The type of packaging will generally depend on the route of administration. Implantable sustained release formulations, as well as transdermal formulations, are also contemplated.
Some examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to: ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffer substances (e.g., phosphates), glycine, sorbic acid or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts), colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene block copolymers, lanolin, sugars (e.g., lactose, glucose and sucrose), starches (e.g., corn starch and potato starch), cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; tragacanth powder; malt; gelatin; talc powder; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols, such as propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; ringer's solution; ethanol; and phosphate buffers, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate. Coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preserving and anti-oxidizing agents may also be present in the composition, according to the judgment of the person skilled in the art.
The compounds of the present invention may be used alone or in combination with other therapeutic agents for the treatment of the diseases or conditions described herein, such as cancer. In certain embodiments, the compounds of the present invention are combined in a pharmaceutical combination formulation, or as a combination therapy in a dosing regimen, with a second compound having anti-hyperproliferative properties or for the treatment of a hyperproliferative disease, such as cancer. The second compound of the pharmaceutical combination or dosing regimen preferably has complementary activities to the compounds of the present invention such that they do not adversely affect each other. Such compounds are suitably present in the combination in an amount effective for the intended purpose. In one embodiment, the compounds of the present invention are combined with other anti-tumor agents. The anti-tumor medicine comprises: alkylating agents including, but not limited to, cyclophosphamide, mechlorethamine, melphalan, cinchonine, carmustine; platinoids including but not limited to carboplatin, cisplatin, oxaliplatin; topoisomerase inhibitors including, but not limited to, topotecan, camptothecin, topotecan, irinotecan; antibiotics, including but not limited to, anthracyclines, actinomycin D, daunorubicin, doxorubicin, mitoxantrone, bleomycin, and plicamycin; anti-microtubule or anti-mitotic agents including, but not limited to, paclitaxel, vinorelbine, docetaxel, doxorubicin; antimetabolites including, but not limited to, fluorouracil, methotrexate, cytarabine, mercaptopurine (mecaptoprine), thioguanine, and gemcitabine; antibodies, including but not limited to herceptin, bevacizumab; hormones, including but not limited to letrozole (letrozole), vorozole (vorazole), tamoxifen, toremifene, fulvestrant, flutamide, nilutamide, triptorelin; kinase inhibitors, EGFR kinase inhibitors, including but not limited to gefitinib (gefitinib), erlotinib (erlotinib), lapatinib (lapatinib), afatinib (afatinib); VEGFR inhibitors including, but not limited to, Sorafenib (Sorafenib), regrafenib (Regorafenib), Sunitinib (Sunitinib), Cabozantinib (Cabozantinib), Pazopanib (Pazopanib), vandetanib (vandetanib), axitinib (axitinib); ALK inhibitors including, but not limited to, Crizotinib (Crizotinib), ceritinib (ceritinib), Alectinib; Bcr-Abl inhibitors, including but not limited to Imatinib (Imatinib), panatinib (Ponatinib), Nilotinib (Nilotinib), Dasatinib (Dasatinib); BTK inhibitors, including but not limited to Ibrutinib (Ibrutinib); B-RAF inhibitors, including but not limited to Vemurafenib (Vemurafenib); cyclin-dependent kinase CDK4/6 inhibitors, Palbociclib (Palbociclib); mTOR inhibitors, including but not limited to rapamycin (rapamycin), everolimus (everolimus); sirtuin inhibitors, including but not limited to vorinostat (vorinostat); PD1/PDL1 antibody, Keytruda (Pembrolizumab), Opdivo (Nivolumab).
In a fourth aspect of the present invention, there is provided a use of the compound of the first aspect, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the third aspect, in the preparation of a medicament for preventing and/or treating a STAT 3-mediated disease, wherein the STAT 3-mediated disease includes cancer, an immune disease, a cardiovascular disease, a viral infection, an inflammatory disease, a metabolic/endocrine dysfunction, or a neurological disease.
The beneficial technical effects are as follows: the compound has strong in-vitro antiproliferative activity on human prostate cancer cell DU145 with high STAT3 expression. The compound shows obvious inhibition activity on STAT3 phosphorylation of human prostate cancer DU145 in an immunoblotting experiment. In vivo pharmacodynamic studies show that the compound of the invention shows a remarkable growth inhibition effect in a nude mouse human prostate cancer DU145 xenograft model.
Drawings
FIG. 1 is an image of the immunoblot results showing the inhibition of p-STAT3 expression by human prostate cancer cells DU145 cells of examples 1-7, 12, 13 and 15.
FIG. 2 is a tumor growth curve showing the growth inhibition of example 1 in a human prostate cancer DU145 xenograft model in nude mice.
Detailed Description
The following are specific examples of the present invention, which further describe the technical solutions of the present invention, but the scope of the present invention is not limited to these examples. All changes which come within the meaning and range of equivalency of the claims are to be embraced within their scope.
In the following examples, molecules with a single chiral center exist as a racemic mixture unless otherwise noted by structural formula or chemical name. Unless otherwise noted by structural formula or chemical name, those molecules having two or more chiral centers exist as racemic mixtures of diastereomers. The single enantiomers/diastereomers may be obtained by methods known to those skilled in the art.
Preparation method
The compounds of the present invention may be synthesized according to the synthetic schemes herein and/or by techniques well known in the art. For example, the compounds provided by the present invention can be prepared according to the following general synthetic methods.
In one general synthetic approach, compounds of formula (I) were prepared according to method-1.
Method-1
Figure BDA0001894123760000111
Specifically, in method-1, the pyrazole compound of the present invention can be prepared by 2-step reaction. For example, compound A and compound B are used as starting materials, and compound C is prepared through condensation reaction; and (3) carrying out sulfonylation reaction on the compound C to prepare the pyrazole compound shown in the formula (I).
In another general synthetic method, a compound of formula (I) is prepared according to method-2.
Method-2
Figure BDA0001894123760000112
Specifically, in the general synthetic method-2, the pyrazole compound of the present invention can be prepared by 3-step reaction. For example, compound D is used as a starting material, compound E is prepared by sulfonylation, and it is hydrolyzed with lithium hydroxide to obtain compound F. Compounds F and (1-R)1-1H-pyrazol-4-yl) methylamine or hydrochloride thereof to prepare the pyrazole compound shown in the formula (I).
The compounds of the present invention may be synthesized according to one or more of the synthetic schemes herein and/or by techniques well known in the art. One skilled in the art will recognize that the synthetic methods of certain embodiments described in detail herein can be readily adapted to synthesize other embodiments. In some embodiments, the compounds described herein may be prepared by an appropriate combination of synthetic methods well known in the art. Many starting materials and other reagents are available from commercial suppliers, such as alfa aesar (china) chemical limited, or are readily prepared using synthetic methods commonly used in the art.
1H NMR spectra were recorded on an instrument operating at 400 MHz. The H NMR spectra were obtained as a solution (reported in ppm) using the internal standard tetramethylsilane (0.00ppm) as a reference standard. When peak multiplicities are reported, the following abbreviations are used: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad), dd (doublet of doublets), dt (doublet of triplets). Coupling constants given are in hertz (Hz).
If desired, the (R) -and (S) -isomers of the non-limiting exemplary compounds, if present, can be resolved by methods known to those skilled in the art, e.g., by forming diastereomeric salts or complexes, which can be separated, e.g., by crystallization; by forming diastereomeric derivatives, which can be separated, for example, by crystallization or chromatography; selectively reacting one enantiomer with an enantiomer-specific reagent, followed by separation of the modified and unmodified enantiomers; or chromatographic separation in a chiral environment such as a chiral chromatography column. Alternatively, specific enantiomers may be prepared by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to another by asymmetric conversion.
In the following preparations and examples, "Me" means methyl, "Et" means ethyl, "PE" means petroleum ether, "EtOAc" means ethyl acetate, "MeOH" means methanol, "DMSO-d" means6"refers to deuterated dimethylsulfoxide," DCM "refers to dichloromethane," DMAP "refers to 4-dimethylaminopyridine," HATU "refers to O- (7-azabenzotriazolyl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate," mL "refers to milliliters," mmol "refers to millimoles," μ M "refers to micromoles," nM "refers to nanomoles, and" ° C "refers to degrees Celsius.
Example 1: synthesis of 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000121
Step 1: synthesis of methyl 4-methyl-3- ((4-methylphenyl) sulfonylamino) benzoate
Figure BDA0001894123760000131
A reaction mixture of 3-amino-4-methylbenzoic acid (1.65g, 10mmol), p-toluenesulfonyl chloride (2.8g, 12mmol), pyridine (1.19g, 15mmol) and DMAP (0.122g, 1mmol) in DCM (40mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified with dilute hydrochloric acid to a pH of 3-4. The mixture was extracted with dichloromethane (50 mL. times.3). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel, PE/EtOAc ═ 3:1) to give the product as a yellow oil (2.82g, 88% yield).
1H NMR(400MHz,DMSO-d6)δ9.73(s,1H),7.70–7.63(m,2H),7.55(d,J＝8.0H,2H),7.36(d,J＝8.0Hz,2H),7.28(d,J＝7.7Hz,1H),3.81(s,3H),2.36(s,3H),2.03(s,3H)。
MS(ESI+)m/z 319.8[M+H]+。
Step 2: synthesis of 4-methyl-3- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000132
A mixture of methyl 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoate (2.71g, 8.5mmol) and sodium hydroxide (1.36g, 34mmol) in methanol (51mL) and water (17mL) was refluxed for 7 hours. The resulting mixture was evaporated to dryness under reduced pressure, the reaction mixture was diluted with water (100mL), acidified to pH 2 with concentrated hydrochloric acid, and extracted with ethyl acetate (50mL × 3). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to a solution where solids began to precipitate. The resulting suspension was left to stand for 1 hour, and the resulting solid was collected by suction filtration and dried to give the product as a pale yellow solid (2.1g, 81% yield).
1H NMR(400MHz,DMSO-d6)δ12.89(br s,1H),9.69(br s,1H),7.66(dd,J＝7.8,1.6Hz,1H),7.63(d,J＝1.5Hz,1H),7.55(d,J＝8.2Hz,2H),7.36(d,J＝8.1Hz,2H),7.25(d,J＝7.9Hz,1H),2.36(s,3H),2.05(s,3H)。
MS(ESI+)m/z 305.8[M+H]+。
And step 3: synthesis of 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000133
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.073g, 0.65mmol), HATU (0.285g, 0.75mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 10:1) to give the product as a white solid (111mg, 56% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.73(t,J＝5.7Hz,1H),7.62(d,J＝1.8Hz,1H),7.60–7.56(m,2H),7.52(d,J＝8.3Hz,2H),7.37–7.30(m,3H),7.18(d,J＝8.0Hz,1H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),2.36(s,3H),1.93(s,3H)。
MS(ESI+)m/z399.3[M+H]+。
Example 2: n- ((1-ethyl-1H-pyrazol-4-yl) methyl) -4-methyl-3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000141
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1-ethyl-1H-pyrazol-4-yl) methylamine (0.063g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (115mg, 56% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.73(t,J＝5.7Hz,1H),7.65–7.60(m,2H),7.58(dd,J＝7.9,1.8Hz,1H),7.52(d,J＝8.3Hz,2H),7.37–7.30(m,3H),7.18(d,J＝8.0Hz,1H),4.25(d,J＝5.7Hz,2H),4.07(q,J＝7.3Hz,2H),2.36(s,3H),1.93(s,3H),1.33(t,J＝7.3Hz,3H)。
MS(ESI+)m/z 412.8[M+H]+。
Example 3: 4-methyl-3- ((4-methylphenyl) sulfonylamino) -N- ((1-propyl-1H-pyrazol-4-yl) methyl) -benzamide
Figure BDA0001894123760000151
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1-propyl-1H-pyrazol-4-yl) methylamine (0.070g,0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (83mg, 39% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.73(t,J＝5.7Hz,1H),7.63(d,J＝1.5Hz,1H),7.61–7.56(m,2H),7.52(d,J＝8.3Hz,2H),7.36–7.30(m,3H),7.18(d,J＝8.0Hz,1H),4.25(d,J＝5.7Hz,2H),3.99(t,J＝7.0Hz,2H),2.36(s,3H),1.93(s,3H),1.80–1.68(m,2H),0.81(t,J＝7.4Hz,3H)。
MS(ESI+)m/z 426.8[M+H]+。
Example 4: n- ((1-isopropyl-1H-pyrazol-4-yl) methyl) -4-methyl-3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000152
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1-isopropyl-1H-pyrazol-4-yl) methylamine (0.070g,0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (127mg, 60% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.73(t,J＝5.7Hz,1H),7.66–7.62(m,2H),7.59(dd,J＝7.9,1.8Hz,1H),7.52(d,J＝8.3Hz,2H),7.37–7.30(m,3H),7.18(d,J＝8.0Hz,1H),4.48–4.40(m,1H),4.26(d,J＝5.7Hz,2H),2.36(s,3H),1.93(s,3H),1.38(d,J＝6.7Hz,6H)。
MS(ESI+)m/z 426.8[M+H]+。
Example 5: 3- ((4-ethylphenyl) sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000161
Step 1: synthesis of methyl 3- ((4-ethylphenyl) sulfonylamino) -4-methylbenzoate
Figure BDA0001894123760000162
A reaction mixture of methyl 3-amino-4-methylbenzoate ((1.652g, 10mmol), 4-ethylbenzenesulfonyl chloride (2.456g, 12mmol), pyridine (1.186g, 15mmol) and DMAP (0.244g, 2mmol) in DCM (50mL) was stirred at room temperature overnight, the reaction mixture was diluted with water (100mL), acidified to pH 3 with dilute hydrochloric acid, the mixture was extracted with dichloromethane (50mL × 3), the combined organic layers were washed with water (50mL × 2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 5:1 then 3:1) to give the product as a yellow solid (2.994g, 90% yield).
1H NMR(400MHz,DMSO-d6)δ9.71(s,1H),7.67(dd,J＝7.9,1.7Hz,1H),7.60(d,J＝1.6Hz,1H),7.56(d,J＝8.2Hz,2H),7.39(d,J＝8.1Hz,2H),7.28(d,J＝8.0Hz,1H),3.80(s,3H),2.67(q,J＝7.6Hz,2H),2.04(s,3H),1.17(t,J＝7.6Hz,3H)。
MS(ESI+)m/z 333.8[M+H]+。
Step 2: synthesis of 3- ((4-ethylphenyl) sulfonylamino) -4-methylbenzoic acid
Figure BDA0001894123760000163
A mixture of methyl 3- ((4-ethylphenyl) sulfonylamino) -4-methylbenzoate (2.2g, 6.6mmol), lithium hydroxide monohydrate (1.109g, 26mmol), tetrahydrofuran (13mL), methanol (3.3mL) and water (6.6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (150mL), acidified to pH 2 with concentrated hydrochloric acid and extracted with ethyl acetate (100mL × 3). The combined organic layers were washed with water (100mL) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (2.083g, 99% yield).
1H NMR(400MHz,DMSO-d6)δ12.86(br s,1H),9.65(br s,1H),7.65(dd,J＝7.9,1.8Hz,1H),7.60–7.53(m,3H),7.39(d,J＝8.3Hz,2H),7.25(d,J＝8.0Hz,1H),2.66(q,J＝7.6Hz,2H),2.04(s,3H),1.17(t,J＝7.6Hz,3H)。
MS(ESI+)m/z 320.0[M+H]+。
And step 3: synthesis of 3- ((4-ethylphenyl) sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000171
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.096g, 0.3mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.043g, 0.39mmol,; HATU (0.171g, 0.45mmol) and triethylamine (0.091g,0.9mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 20:1) to give the product as a white solid (66mg, 53% yield).
1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.73(t,J＝5.6Hz,1H),7.62(d,J＝1.8Hz,1H),7.60-7.52(m,4H),7.37(d,J＝8.5Hz,2H),7.32(d,J＝0.5Hz,1H),7.18(d,J＝8.0Hz,1H),4.24(d,J＝5.6Hz,2H),3.78(s,3H),2.66(q,J＝7.6Hz,2H),1.92(s,3H),1.17(t,J＝7.6Hz,3H).
MS(ESI+)m/z 412.8[M+H]+.
Example 6: 3- ((4-cyclopropylphenyl) sulfonamido) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000172
Step 1: synthesis of methyl 3- ((4-cyclopropylphenyl) sulfonylamino) -4-methylbenzoate
Figure BDA0001894123760000173
A reaction mixture of methyl 3-amino-4-methylbenzoate (0.330g, 2mmol), 4-cyclopropylbenzenesulfonyl chloride (0.455g, 2.1mmol), pyridine (0.206g, 2.6mmol) and DMAP (0.049g, 0.4mmol) in DCM (10mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (30mL), adjusted to pH 2-3 with 1M hydrochloric acid solution, and the aqueous layer was extracted with dichloromethane (30 mL. times.2). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel, DCM/MeOH ═ 200:1) to give the product as a yellow solid (0.572g, 83% yield).
1H NMR(400MHz,DMSO-d6)δ9.69(s,1H),7.66(dd,J＝7.9,1.7Hz,1H),7.60(d,J＝1.7Hz,1H),7.50(d,J＝8.4Hz,2H),7.28(d,J＝8.0Hz,1H),7.23(d,J＝8.4Hz,2H),3.80(s,3H),2.03(s,3H),2.02–1.94(m,1H),1.12–0.96(m,2H),0.81–0.68(m,2H)。
MS(ESI+)m/z 345.8[M+H]+。
Step 2: synthesis of 3- ((4-cyclopropylphenyl) sulfonylamino) -4-methylbenzoic acid
Figure BDA0001894123760000181
A mixture of methyl 3- ((4-cyclopropylphenyl) sulfonylamino) -4-methylbenzoate (0.570g, 1.65mmol) and lithium hydroxide monohydrate (0.174g, 4.2mmol), tetrahydrofuran (4mL), methanol (0.75mL) and water (1.5mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (50mL), and the aqueous layer was acidified with concentrated hydrochloric acid to pH 2 and extracted with ethyl acetate (30mL × 3). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (0.496g, 91% yield).
1H NMR(400MHz,DMSO-d6)δ12.85(s,1H),9.63(s,1H),7.64(dd,J＝7.9,1.7Hz,1H),7.58(d,J＝1.7Hz,1H),7.50(d,J＝8.4Hz,2H),7.29–7.19(m,3H),2.04(s,3H),2.08–1.94(m,1H),1.08–1.00(m,2H),0.78–0.70(m,2H)。
MS(ESI+)m/z 331.8[M+H]+。
And step 3: synthesis of 3- ((4-cyclopropylphenyl) sulfonamido) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000182
A reaction mixture of 4-methyl-3- ((4-cyclopropylphenyl) sulfonamido) benzoic acid (0.099g, 0.3mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.054g,0.39mmol), HATU (0.171g, 0.45mmol) and triethylamine (0.091g,0.9mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 10:1) to give the product as a white solid (107mg, 84% yield).
1H NMR(400MHz,DMSO-d6)δ9.59(s,1H),8.73(t,J＝5.7Hz,1H),7.62(d,J＝1.8Hz,1H),7.60–7.54(m,2H),7.49(d,J＝8.4Hz,2H),7.33(d,J＝0.5Hz,1H),7.21(d,J＝8.4Hz,2H),7.17(d,J＝8.1Hz,1H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),2.04–1.94(m,1H),1.93(s,3H),1.10–0.96(m,2H),0.79–0.67(m,2H)。
MS(ESI+)m/z 424.8[M+H]+。
Example 7: 3- ((4-methoxyphenyl) sulfonamide) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000191
Step 1: synthesis of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000192
A reaction mixture of 3-amino-4-methylbenzoic acid (1.512g, 10mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (1.112g,10mmol), HATU (4.562g, 12mmol) and triethylamine (3.04g,30mmol) in DCM (100mL) was stirred overnight. Water (150mL) was added, and the resulting mixture was extracted with DCM (100 mL. times.3). the organic layers were combined, washed with water (100 mL. times.1), washed with brine (100 mL. times.1), dried, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, DCM/MeOH ═ 30:1) to give the product as a white solid (1.482g, 67% yield).
1H NMR(400MHz,DMSO-d6)δ8.47(t,J＝5.7Hz,1H),7.55(s,1H),7.32(d,J＝0.4Hz,1H),7.09(d,J＝1.4Hz,1H),6.98–6.90(m,2H),4.96(s,2H),4.22(d,J＝5.8Hz,2H),3.77(s,3H),2.07(s,3H)。
MS(ESI+)m/z 244.9[M+H]+。
Step 2: synthesis of 3- ((4-methoxyphenyl) sulfonamide) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000201
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), p-methoxysulfonyl chloride (0.079g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (79mg, 64% yield).
1H NMR(400MHz,DMSO-d6)δ9.53(s,1H),8.73(t,J＝5.7Hz,1H),7.62(d,J＝1.8Hz,1H),7.59–7.53(m,4H),7.33(d,J＝0.4Hz,1H),7.18(d,J＝8.1Hz,1H),7.05(d,J＝9.0Hz,2H),4.24(d,J＝5.7Hz,2H),3.81(s,3H),3.78(s,3H),1.95(s,3H)。
MS(ESI+)m/z 414.8[M+H]+。
Example 8: 3- ((4-ethoxyphenyl) sulfonamido) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000202
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 4-ethoxybenzenesulfonyl chloride (0.079g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (91mg, 71% yield).
1H NMR(400MHz,DMSO-d6)δ9.52(s,1H),8.73(t,J＝5.7Hz,1H),7.62(d,J＝1.7Hz,1H),7.60–7.51(m,4H),7.33(s,1H),7.18(d,J＝8.0Hz,1H),7.03(d,J＝8.9Hz,2H),4.24(d,J＝5.7Hz,2H),4.08(q,J＝7.0Hz,2H),3.78(s,3H),1.95(s,3H),1.33(t,J＝7.0Hz,3H)。
MS(ESI+)m/z 428.8[M+H]+。
Example 9: 3- ((4-isopropylphenyl) sulfonamide) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000211
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 4-isopropylbenzenesulfonyl chloride (0.079g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (81mg, 63% yield).
1H NMR(400MHz,DMSO-d6)δ9.60(s,1H),8.73(t,J＝5.8Hz,1H),7.64–7.50(m,5H),7.40(d,J＝8.4Hz,2H),7.32(s,1H),7.18(d,J＝8.0Hz,1H),4.23(d,J＝5.7Hz,2H),3.78(s,3H),3.04–2.87(m,1H),1.91(s,3H),1.19(d,J＝6.9Hz,6H)。
MS(ESI+)m/z 426.8[M+H]+。
Example 10: 3- ((4-chlorophenyl) sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000212
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 4-chlorobenzenesulfonyl chloride (0.076g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (92mg, 73% yield).
1H NMR(400MHz,DMSO-d6)δ9.83(s,1H),8.75(t,J＝5.6Hz,1H),7.65–7.55(m,7H),7.33(s,1H),7.21(d,J＝8.0Hz,1H),4.24(d,J＝5.6Hz,2H),3.78(s,3H),1.96(s,3H)。
MS(ESI+)m/z 418.7[M+H]+。
Example 11: 4-Ethyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000221
Step 1: synthesis of 3-amino-4-ethyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000222
A reaction mixture of 3-amino-4-ethylbenzoic acid (0.330g, 2.0mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.222g, 2.0mmol), HATU (0.912g, 2.4mmol) and triethylamine (0.607g, 6.0mmol) in DCM (20mL) was stirred overnight. Water (40mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel, DCM/MeOH ═ 70:1 then 50:1) to give the product as a yellow solid (0.482g, 93% yield).
MS(ESI+)m/z 259.2[M+H]+。
Step 2: synthesis of 4-ethyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000223
A mixture of 3-amino-4-ethyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.129g, 0.5mmol), p-methylbenzenesulfonyl chloride (0.114g, 0.6mmol), pyridine (0.059g, 0.75mmol) and DMAP (0.012g, 0.1mmol) in DCM (10mL) was stirred at room temperature overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (0.171g, 83% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.72(t,J＝5.7Hz,1H),7.62(dd,J＝8.0,1.8Hz,1H),7.58–7.50(m,4H),7.37–7.29(m,3H),7.23(d,J＝8.1Hz,1H),4.23(d,J＝5.7Hz,2H),3.78(s,3H),2.42(q,J＝7.5Hz,2H),2.36(s,3H),0.91(t,J＝7.5Hz,3H)。
MS(ESI+)m/z 413.2[M+H]+。
Example 12: n- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000231
Step 1: synthesis of methyl 3- ((4-methylphenyl) sulfonylamino) benzoate
Figure BDA0001894123760000232
A reaction mixture of methyl 3-aminobenzoate (0.756g, 5mmol), p-toluenesulfonyl chloride (1.144g, 6mmol), pyridine (0.593g, 7.5mmol) and DMAP (0.122g, 1mmol) in DCM (20mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with dichloromethane (50 mL. times.2). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 2:1) to give the product as a white solid (1.511g, 99% yield).
1H NMR(400MHz,DMSO-d6)δ10.51(s,1H),7.75–7.72(m,1H),7.66(d,J＝8.3Hz,2H),7.62–7.58(m,1H),7.41–7.37(m,2H),7.35(d,J＝8.1Hz,2H),3.82(s,3H),2.32(s,3H).MS(ESI+)m/z 306.1[M+H]+。
Step 2: synthesis of 3- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000233
A mixture of methyl-3- ((4-methylphenyl) sulfonylamino) benzoate (1.500g, 4.9mmol), lithium hydroxide (0.821g, 19.5mmol), tetrahydrofuran (15mL), methanol (3mL) and water (6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL), acidified to pH 2 with concentrated hydrochloric acid and extracted with ethyl acetate (50mL × 2). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (1.431g, 97% yield).
1H NMR(400MHz,DMSO-d6)δ13.02(s,1H),10.45(s,1H),7.71–7.68(m,1H),7.64(d,J＝8.3Hz,2H),7.60–7.56(m,1H),7.39–7.30(m,4H),2.33(s,3H)。
MS(ESI+)m/z 292.1[M+H]+。
And step 3: synthesis of N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000241
A reaction mixture of 3- ((4-methylphenyl) sulfonamido) benzoic acid (0.146g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.60mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (114mg, 59% yield).
1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.76(t,J＝5.7Hz,1H),7.64(d,J＝8.0Hz,2H),7.58(t,J＝1.9Hz,1H),7.56(s,1H),7.49–7.45(m,1H),7.33(d,J＝8.0Hz,2H),7.32(s,1H),7.28(t,J＝7.8Hz,1H),7.22(ddd,J＝8.1,2.2,1.0Hz,1H),4.23(d,J＝5.7Hz,2H),3.77(s,3H),2.32(s,3H)。
MS(ESI+)m/z 385.1[M+H]+。
Example 13: 3-chloro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -5- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000242
Step 1: synthesis of methyl 3-chloro-5- ((4-methylphenyl) sulfonylamino) benzoate
Figure BDA0001894123760000243
A reaction mixture of methyl 3-amino-5-chlorobenzoate (0.928g, 5mmol), p-toluenesulfonyl chloride (1.144g, 6mmol), pyridine (0.593g, 7.5mmol) and DMAP (0.122g, 1mmol) in DCM (20mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with dichloromethane (50 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 3:1) to give the product as a yellow solid (1.649g, 97% yield).
1H NMR(400MHz,DMSO-d6)δ10.81(s,1H),7.71–7.66(m,3H),7.55(t,J＝2.0Hz,1H),7.41–7.36(m,3H),3.83(s,3H),2.34(s,3H)。
MS(ESI+)m/z 340.0[M+H]+。
Step 2: synthesis of 3-chloro-5- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000251
A mixture of methyl 3-chloro-5- ((4-methylphenyl) sulfonamido) benzoate (1.600g, 4.71mmol), lithium hydroxide (0.791g, 19mmol), tetrahydrofuran (15mL), methanol (3mL) and water (6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL), acidified to pH 2 with concentrated HCl and extracted with ethyl acetate (50 mL. times.2). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (1.54g, 100% yield).
1H NMR(400MHz,DMSO-d6)δ13.44(br s,1H),10.77(br s,1H),7.68(d,J＝8.3Hz,2H),7.65(dd,J＝2.0,1.6Hz,1H),7.55–7.50(m,1H),7.39(d,J＝8.1Hz,2H),7.34(t,J＝2.0Hz,1H),2.34(s,3H)。
MS(ESI+)m/z 326.0[M+H]+。
And step 3: synthesis of 3-chloro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -5- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000252
A reaction mixture of 3-chloro-5- ((4-methylphenyl) sulfonamido) benzoic acid (0.163g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (0.120g, 57% yield).
1H NMR(400MHz,DMSO-d6)δ10.67(s,1H),8.88(t,J＝5.7Hz,1H),7.67(d,J＝8.3Hz,2H),7.60–7.52(m,3H),7.37(d,J＝8.1Hz,2H),7.32(s,1H),7.24(t,J＝2.0Hz,1H),4.23(d,J＝5.6Hz,2H),3.77(s,3H),2.34(s,3H)。
MS(ESI+)m/z 419.1[M+H]+。
Example 14: 3-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -5- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000261
Step 1: synthesis of methyl 3-methyl-5- ((4-methylphenyl) sulfonylamino) benzoate
Figure BDA0001894123760000262
A reaction mixture of methyl 3-amino-5-methylbenzoate (0.826g, 5mmol), p-toluenesulfonyl chloride (1.144g, 6mmol), pyridine (0.593g, 7.5mmol) and DMAP (0.122g, 1mmol) in DCM (20mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with dichloromethane (50 mL. times.2). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 2:1) to give the product as a white solid (1.579g, 99% yield).
1H NMR(400MHz,DMSO-d6)δ10.43(s,1H),7.65(d,J＝8.3Hz,2H),7.53(s,1H),7.43(s,1H),7.35(d,J＝8.1Hz,2H),7.18(s,1H),3.80(s,3H),2.32(s,3H),2.26(s,3H)。
MS(ESI+)m/z 320.1[M+H]+。
Step 2: synthesis of 3-methyl-5- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000263
A mixture of methyl 3-methyl-5- ((4-methylphenyl) sulfonamido) benzoate (1.550g,4.85mmol), lithium hydroxide (0.815g,19.4mmol), tetrahydrofuran (15mL), methanol (3mL) and water (6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL), acidified to pH 2 with concentrated hydrochloric acid and extracted with ethyl acetate (50mL × 2). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (1.492g, 100% yield).
1H NMR(400MHz,DMSO-d6) δ 12.96(s,1H),10.39(s,1H),7.66(d, J ═ 8.3Hz,2H),7.51(t, J ═ 1.3Hz,1H),7.41(s,1H),7.35(d, J ═ 8.0Hz,2H),7.16(s,1H),2.33(s,3H),2.26(s, 3H). And step 3: 3-methyl-N- ((1-methyl-1H-pyrazol-4-yl)) Synthesis of methyl) -5- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000271
A reaction mixture of 3-methyl-5- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (0.107g, 54% yield).
1H NMR(400MHz,DMSO-d6)δ10.28(s,1H),8.68(t,J＝5.7Hz,1H),7.64(d,J＝8.3Hz,2H),7.55(s,1H),7.38(s,1H),7.36–7.28(m,4H),7.04(s,1H),4.21(d,J＝5.7Hz,2H),3.77(s,3H),2.32(s,3H),2.23(s,3H)。
MS(ESI+)m/z 399.1[M+H]+。
Example 15: 3-fluoro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -5- ((4-methylphenyl) sulfonamido) benzamide
Figure BDA0001894123760000272
Step 1: synthesis of methyl 3-fluoro-5- ((4-methylphenyl) sulfonylamino) benzoate
Figure BDA0001894123760000273
A reaction mixture of methyl 3-amino-5-fluorobenzoate (0.846g, 5mmol), p-toluenesulfonyl chloride (1.144g, 6mmol), pyridine (0.593g, 7.5mmol) and DMAP (0.122g, 1mmol) in DCM (20mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with dichloromethane (50 mL. times.2). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 4:1) to give the product as a white solid (1.566g, 97% yield).
1H NMR(400MHz,DMSO-d6)δ10.82(s,1H),7.70(d,J＝8.3Hz,2H),7.57–7.55(m,1H),7.38(d,J＝8.1Hz,2H),7.35–7.31(m,1H),7.21–7.16(m,1H),3.83(s,3H),2.34(s,3H)。
MS(ESI+)m/z 324.1[M+H]+。
Step 2: synthesis of 3-fluoro-5- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000281
A mixture of methyl 3-fluoro-5- ((4-methylphenyl) sulfonamido) benzoate (1.550g,4.8 mmol), lithium hydroxide (0.805g, 19mmol), tetrahydrofuran (15mL), methanol (3mL) and water (6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL), acidified to pH 2 with concentrated hydrochloric acid and extracted with ethyl acetate (50mL × 2). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (1.465g, 99% yield).
1H NMR(400MHz,DMSO-d6)δ13.38(br s,1H),10.78(br s,1H),7.69(d,J＝8.3Hz,2H),7.55–7.49(d,J＝1.6Hz,1H),7.38(d,J＝8.1Hz,2H),7.29(ddd,J＝8.9,2.4,1.3Hz,1H),7.15(dt,J＝10.4,2.3Hz,1H),2.34(s,3H)。
MS(ESI+)m/z 310.0[M+H]+。
And step 3: synthesis of 3-fluoro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -5- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000282
A reaction mixture of 3-fluoro-5- ((4-methylphenyl) sulfonamido) benzoic acid (0.155g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (0.134g, 67% yield).
1H NMR(400MHz,DMSO-d6)δ10.68(s,1H),8.83(t,J＝5.7Hz,1H),7.68(d,J＝8.3Hz,2H),7.57(s,1H),7.42(t,J＝1.6Hz,1H),7.37(d,J＝8.0Hz,2H),7.34–7.29(m,2H),7.04(dt,J＝10.4,2.2Hz,1H),4.23(d,J＝5.6Hz,2H),3.77(s,3H),2.33(s,3H)。
MS(ESI+)m/z 403.1[M+H]+。
Example 16: 4-fluoro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonamido) benzamide
Figure BDA0001894123760000291
Step 1: synthesis of methyl 4-fluoro-3- ((4-methylphenyl) sulfonylamino) benzoate
Figure BDA0001894123760000292
A reaction mixture of methyl 3-amino-4-fluorobenzoate (0.846g, 5mmol), p-toluenesulfonyl chloride (1.144g, 6mmol), pyridine (0.593g, 7.5mmol) and DMAP (0.122g, 1mmol) in DCM (20mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with dichloromethane (50 mL. times.2). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 4:1) to give the product as a white solid (1.592g, 98% yield).
1H NMR(400MHz,DMSO-d6)δ10.39(s,1H),7.89(dd,J＝7.7,2.2Hz,1H),7.81–7.74(m,1H),7.62(d,J＝8.3Hz,2H),7.37(d,J＝8.4Hz,2H),7.31(dd,J＝10.1,8.6Hz,1H),3.84(s,3H),2.36(s,3H)。
MS(ESI+)m/z 324.1[M+H]+。
Step 2: synthesis of 4-fluoro-3- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000293
A mixture of methyl 4-fluoro-3- ((4-methylphenyl) sulfonamido) benzoate (1.580g, 4.9mmol), lithium hydroxide (0.821g, 19.5mmol), tetrahydrofuran (15mL), methanol (3mL) and water (6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL), acidified to pH 2 with concentrated HCl and extracted with ethyl acetate (50 mL. times.2). The combined organic layers were washed with water (100mL × 2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (1.498g, 97% yield).
1H NMR(400MHz,DMSO-d6)δ13.13(br s,1H),10.32(br s,1H),7.85(dd,J＝7.7,2.1Hz,1H),7.77–7.70(m,1H),7.61(d,J＝8.3Hz,2H),7.36(d,J＝8.1Hz,2H),7.28(dd,J＝10.1,8.6Hz,1H),2.36(s,3H)。
MS(ESI+)m/z[M+H]+。
And step 3: synthesis of 4-fluoro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000301
A reaction mixture of 4-fluoro-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.155g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (0.097g, 48% yield).
1H NMR(400MHz,DMSO-d6)δ10.24(s,1H),8.86(t,J＝5.7Hz,1H),7.84(dd,J＝7.7,2.2Hz,1H),7.68(ddd,J＝8.5,4.5,2.2Hz,1H),7.63–7.54(m,3H),7.37–7.31(m,3H),7.23(dd,J＝10.0,8.7Hz,1H),4.25(d,J＝5.6Hz,2H),3.78(s,3H),2.35(s,3H)。
MS(ESI+)m/z 403.1[M+H]+。
Example 17: 4-chloro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000302
Step 1: 4-chloro-3- ((4-methylphenyl) sulfonylamino) benzoic acid methyl ester
Figure BDA0001894123760000311
A reaction mixture of methyl 3-amino-4-chlorobenzoate (0.928g, 5mmol), p-toluenesulfonyl chloride (1.144g, 6mmol), pyridine (0.593g, 7.5mmol) and DMAP (0.122g, 1mmol) in DCM (20mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL) and acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with dichloromethane (50 mL. times.2). The combined organic layers were washed with water (50 mL. times.2) and brine (50mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (silica gel, PE/EtOAc ═ 2:1) to give the product as a white solid (1.226g, 72% yield).
1H NMR(400MHz,DMSO-d6)δ10.19(s,1H),7.86(d,J＝2.0Hz,1H),7.73(dd,J＝8.4,2.1Hz,1H),7.61(d,J＝8.3Hz,2H),7.56(d,J＝8.4Hz,1H),7.37(d,J＝8.1Hz,2H),3.85(s,3H),2.37(s,3H).
MS(ESI+)m/z 339.9[M+H]+.
Step 2: synthesis of 4-chloro-3- ((4-methylphenyl) sulfonylamino) benzoic acid
Figure BDA0001894123760000312
A mixture of methyl 4-chloro-3- ((4-methylphenyl) sulfonamido) benzoate (1.226g, 3.6mmol), lithium hydroxide (0.605g, 14.4mmol), tetrahydrofuran (15mL), methanol (3mL) and water (6mL) was stirred at room temperature overnight. The reaction mixture was diluted with water (100mL), acidified to pH 3 with concentrated hydrochloric acid and extracted with ethyl acetate (50mL × 2). The combined organic layers were washed with water (100 mL. times.2) and brine (100mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give the product as a white solid (1.057g, 90% yield).
1H NMR(400MHz,DMSO-d6)δ13.26(br s,1H),10.18(br s,1H),7.85(d,J＝2.0Hz,1H),7.72(dd,J＝8.3,2.1Hz,1H),7.61(d,J＝8.3Hz,2H),7.53(d,J＝8.4Hz,1H),7.37(d,J＝8.2Hz,2H),2.37(s,3H).
And step 3: synthesis of 4-chloro-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000313
A reaction mixture of 4-chloro-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.155g, 0.5mmol), (1-methyl-1H-pyrazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (0.103g, 49% yield).
1H NMR(400MHz,DMSO-d6)δ10.05(s,1H),8.91(t,J＝5.7Hz,1H),7.84(d,J＝2.0Hz,1H),7.66(dd,J＝8.4,2.1Hz,1H),7.61–7.55(m,3H),7.48(d,J＝8.4Hz,1H),7.38–7.31(m,3H),4.25(d,J＝5.6Hz,2H),3.78(s,3H),2.36(s,3H)。
MS(ESI+)m/z 419.1[M+H]+。
Example 18: 3- ((6-methoxypyridine) -3-sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000321
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 6-methoxypyridine-3-sulfonyl chloride (0.075g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, EtOAc/MeOH ═ 10:1) to give the product as a white solid (106mg, 85% yield).
1H NMR(400MHz,DMSO-d6)δ9.79(s,1H),8.76(t,J＝5.7Hz,1H),8.36(d,J＝2.5Hz,1H),7.88(dd,J＝8.8,2.6Hz,1H),7.62(dd,J＝7.9,1.8Hz,1H),7.57(d,J＝2.0Hz,2H),7.32(s,1H),7.23(d,J＝8.0Hz,1H),6.98(dd,J＝8.8,0.4Hz,1H),4.24(d,J＝5.7Hz,2H),3.91(s,3H),3.78(s,3H),2.03(s,3H)。
MS(ESI+)m/z 415.8[M+H]+。
Example 19: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((6-methylpyridine) -3-sulfonamido) benzamide
Figure BDA0001894123760000331
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 6-methylpyridine-3-sulfonyl chloride (0.069g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, EtOAc/MeOH ═ 10:1) to give the product as a white solid (88mg, 73% yield).
1H NMR(400MHz,DMSO-d6)δ9.85(s,1H),8.72(t,J＝5.7Hz,1H),8.58(d,J＝2.2Hz,1H),7.83(dd,J＝8.2,2.5Hz,1H),7.58(dd,J＝7.9,1.8Hz,1H),7.53(s,2H),7.40(d,J＝8.2Hz,1H),7.28(s,1H),7.18(d,J＝8.0Hz,1H),4.20(d,J＝5.7Hz,2H),3.74(s,3H),2.50(s,3H),1.94(s,3H)。
MS(ESI+)m/z 399.8[M+H]+。
Example 20: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((5-methylthiophene) -2-sulfonylamino) benzamide
Figure BDA0001894123760000332
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 5-methylthiophene-2-sulfonyl chloride (0.070g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (100mg, 83% yield).
1H NMR(400MHz,DMSO-d6)δ9.81(s,1H),8.75(t,J＝5.7Hz,1H),7.66(d,J＝1.8Hz,1H),7.62(dd,J＝8.0,1.8Hz,1H),7.58(s,1H),7.30(s,1H),7.23(d,J＝8.0Hz,1H),7.18(d,J＝3.6Hz,1H),6.84(d,J＝3.6Hz,1H),4.25(d,J＝5.7Hz,2H),3.78(s,3H),2.46(s,3H),2.03(s,3H)。
MS(ESI+)m/z 404.8[M+H]+。
Example 21: 3- ((2, 3-dihydrobenzo [ b ] [1,4] dioxinyl) -6-sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000341
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 2, 3-dihydrobenzo [ b ] [1,4] dioxin-6-sulfonyl chloride (0.084g, 0.36mmol), DMAP (0.007g, 0.06mmol), and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, EtOAc/MeOH ═ 10:1) to give the product as a white foamy solid (105mg, 79% yield).
1H NMR(400MHz,DMSO-d6)δ9.54(s,1H),8.73(t,J＝5.6Hz,1H),7.60–7.55(m,3H),7.32(s,1H),7.20(d,J＝8.9Hz,1H),7.11–7.06(m,2H),7.01–6.95(m,1H),4.33–4.22(m,6H),3.78(s,3H),2.00(s,3H)。
MS(ESI+)m/z 442.8[M+H]+。
Example 22: 3- ((2, 3-dihydro-1H-indene) -5-sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000342
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.095g, 0.39mmol), 2, 3-dihydro-1H-indene-5-sulfonyl chloride (0.102g, 0.47mmol), DMAP (0.010g, 0.08mmol) and pyridine (0.047g, 0.59mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, EtOAc/MeOH ═ 10:1) to give the product as a white solid (70mg, 55% yield).
1H NMR(400MHz,DMSO-d6)δ9.56(s,1H),8.73(t,J＝5.7Hz,1H),7.60(d,J＝1.7Hz,1H),7.59–7.55(m,2H),7.48(s,1H),7.40(dd,J＝7.9,1.7Hz,1H),7.35(d,J＝8.0Hz,1H),7.32(s,1H),7.19(d,J＝8.0Hz,1H),4.23(d,J＝5.7Hz,2H),3.78(s,3H),2.90(t,J＝7.5Hz,2H),2.85(t,J＝7.5Hz,2H),2.10–1.99(m,2H),1.98(d,J＝5.9Hz,3H)。
Example 23: 3- (benzo [ d ] [1,3] dioxol-5-sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000351
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), benzo [ d ] [1,3] dioxol-5-sulfonyl chloride (0.079g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, EtOAc/MeOH ═ 10:1) to give the product as a white solid (113mg, 88% yield).
1H NMR(400MHz,DMSO-d6)δ9.57(s,1H),8.74(t,J＝5.7Hz,1H),7.63–7.53(m,3H),7.32(d,J＝0.6Hz,1H),7.21(dd,J＝8.4,0.6Hz,1H),7.14(dd,J＝8.2,1.9Hz,1H),7.09(d,J＝1.8Hz,1H),7.02(d,J＝8.2Hz,1H),6.15(s,2H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),2.01(s,3H)。
MS(ESI+)m/z 428.8[M+H]+。
Example 24: 3- ((2, 3-dihydrobenzofuranyl) -5-sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000352
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.3mmol), 2, 3-dihydrobenzofuran-5-sulfonyl chloride (0.079g, 0.36mmol), DMAP (0.007g, 0.06mmol) and pyridine (0.036g, 0.45mmol) in DCM (5mL) was stirred overnight. Water (25mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (50 mL. times.2) and brine (50 mL. times.2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, EtOAc/MeOH ═ 10:1) to give the product as a white solid (90mg, 70% yield).
1H NMR(400MHz,DMSO-d6)δ9.46(s,1H),8.73(t,J＝5.7Hz,1H),7.58(dt,J＝7.8,2.5Hz,3H),7.50–7.46(m,1H),7.38(dd,J＝8.5,2.1Hz,1H),7.32(s,1H),7.19(d,J＝7.9Hz,1H),6.85(d,J＝8.4Hz,1H),4.62(t,J＝8.8Hz,2H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),3.18(t,J＝8.8Hz,2H),2.00(s,3H)。
MS(ESI+)m/z 426.5[M+H]+。
Comparative example 1: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- (phenylsulfonylamino) benzamide
Figure BDA0001894123760000361
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), benzenesulfonyl chloride (0.064g, 0.36mmol), pyridine (0.036g, 0.45mmol), and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (94mg, 82% yield).
1H NMR(400MHz,DMSO-d6)δ9.70(s,1H),8.73(t,J＝5.7Hz,1H),7.67–7.50(m,8H),7.32(s,1H),7.18(d,J＝7.7Hz,1H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),1.92(s,3H)。
MS(ESI+)m/z 384.8[M+H]+。
Comparative example 2: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((3-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000362
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), 3-methylbenzenesulfonyl chloride (0.069g, 0.36mmol), pyridine (0.036g, 0.45mmol), and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, ethyl acetate/methanol ═ 10:1) to give the product as a white solid (83mg, 69% yield).
1H NMR(400MHz,DMSO-d6)δ9.64(s,1H),8.73(t,J＝5.7Hz,1H),7.61–7.54(m,3H),7.48–7.38(m,4H),7.32(s,1H),7.19(d,J＝8.4Hz,1H),4.23(d,J＝5.7Hz,2H),3.78(s,3H),2.32(s,3H),1.95(s,3H)。
MS(ESI+)m/z 398.8[M+H]+。
Comparative example 3: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((2-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000371
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), 2-methylbenzenesulfonyl chloride (0.069g, 0.36mmol), pyridine (0.036g, 0.45mmol), and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, ethyl acetate/methanol ═ 10:1) to give the product as a white solid (104mg, 87% yield).
1H NMR(400MHz,DMSO-d6)δ9.72(s,1H),8.71(t,J＝5.7Hz,1H),7.62(dd,J＝7.9,1.3Hz,1H),7.59–7.54(m,3H),7.50(td,J＝7.5,1.4Hz,1H),7.39(d,J＝7.5Hz,1H),7.31(d,J＝0.4Hz,1H),7.31–7.25(m,1H),7.18(d,J＝7.5Hz,1H),4.23(d,J＝5.7Hz,2H),3.78(s,3H),2.54(s,3H),1.99(s,3H)。
MS(ESI+)m/z 398.8[M+H]+。
Comparative example 4: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4-propylphenyl) sulphonamido) benzamide
Figure BDA0001894123760000372
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), 4-propylbenzenesulfonyl chloride (0.079g, 0.36mmol), pyridine (0.036g, 0.45mmol), and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (100mg, 78% yield).
1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.73(t,J＝5.7Hz,1H),7.62(d,J＝1.7Hz,1H),7.60–7.55(m,2H),7.53(d,J＝8.3Hz,2H),7.34(d,J＝8.3Hz,2H),7.32(s,1H),7.17(d,J＝8.0Hz,1H),4.23(d,J＝5.7Hz,2H),3.78(s,3H),2.61(t,J＝7.5Hz,2H),1.90(s,3H),1.64–1.52(m,2H),0.85(t,J＝7.3Hz,3H)。
MS(ESI+)m/z 427.2[M+H]+。
Comparative example 5: 3- ((4-fluorophenyl) sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000381
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), 4-fluorobenzenesulfonyl chloride (0.070g, 0.36mmol), pyridine (0.036g, 0.45mmol), and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (87mg, 72% yield).
1H NMR(400MHz,DMSO-d6)δ9.76(s,1H),8.74(t,J＝5.7Hz,1H),7.73–7.65(m,2H),7.60(dd,J＝7.9,1.8Hz,1H),7.58–7.55(m,2H),7.43–7.36(m,2H),7.32(s,1H),7.21(d,J＝8.0Hz,1H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),1.96(s,3H)。
MS(ESI+)m/z 403.1[M+H]+。
Comparative example 6: 4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) -3- ((4- (trifluoromethyl) phenyl) sulfonamido) benzamide
Figure BDA0001894123760000382
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), 4- (trifluoromethyl) benzenesulfonyl chloride (0.088g, 0.36mmol), pyridine (0.036g, 0.45mmol) and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (94mg, 69% yield).
1H NMR(400MHz,DMSO-d6)δ10.01(s,1H),8.76(t,J＝5.7Hz,1H),7.96(d,J＝8.4Hz,2H),7.85(d,J＝8.3Hz,2H),7.63(dd,J＝7.9,1.7Hz,1H),7.59(s,1H),7.56(s,1H),7.32(s,1H),7.22(d,J＝8.0Hz,1H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),1.94(s,3H)。
MS(ESI+)m/z 453.1[M+H]+。
Comparative example 7: 3- ((4-cyanophenyl) sulfonylamino) -4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000391
A reaction mixture of 3-amino-4-methyl-N- ((1-methyl-1H-pyrazol-4-yl) methyl) benzamide (0.073g, 0.30mmol), 4-cyanobenzenesulfonyl chloride (0.073g, 0.36mmol), pyridine (0.036g, 0.45mmol), and DMAP (0.007g, 0.06mmol) in DCM (5mL) was stirred overnight. The filtrate was diluted with water (30 mL). The resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (20 mL. times.1) and brine (20 mL. times.1), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (74mg, 60% yield).
1H NMR(400MHz,DMSO-d6)δ10.06(s,1H),8.76(t,J＝5.7Hz,1H),8.05(d,J＝8.5Hz,2H),7.80(d,J＝8.5Hz,2H),7.63(dd,J＝7.9,1.8Hz,1H),7.57(s,1H),7.55(d,J＝1.7Hz,1H),7.32(s,1H),7.23(d,J＝8.1Hz,1H),4.24(d,J＝5.7Hz,2H),3.78(s,3H),1.96(s,3H)。
MS(ESI+)m/z 410.1[M+H]+。
Comparative example 8: 4-methyl-N- ((1-methyl-5-oxopyrrolidin-3-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000401
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), 4- (aminomethyl) -1-methylpyrrolidin-2-one (0.084g, 0.65mmol), HATU (0.285g, 0.75mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 10:1) to give the product as a white solid (105mg, 51% yield).
1H NMR(400MHz,CDCl3)δ7.80(d,J＝1.6Hz,1H),7.65(dd,J＝7.9,1.7Hz,1H),7.57(d,J＝8.3Hz,2H),7.36(s,1H),7.31(t,J＝5.7Hz,1H),7.20(d,J＝8.0Hz,2H),7.13(d,J＝8.0Hz,1H),3.65–3.48(m,2H),3.48–3.38(m,1H),3.24(dd,J＝10.1,5.0Hz,1H),2.83(s,3H),2.81–2.71(m,1H),2.57(dd,J＝17.0,9.0Hz,1H),2.38(s,3H),2.34(dd,J＝17.1,5.9Hz,1H),1.94(s,3H)。
MS(ESI+)m/z 416.3[M+H]+。
Comparative example 9: 4-methyl-N- ((1-methyl-1H-pyrazol-5-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000402
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1-methyl-1H-pyrazol-5-yl) methylamine (0.073g, 0.65mmol), HATU (0.285g, 0.75mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (50mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 10:1) to give the product as a white foamy solid (119mg, 60% yield).
1H NMR(400MHz,DMSO-d6)δ9.64(s,1H),8.90(t,J＝5.6Hz,1H),7.65–7.56(m,2H),7.55–7.49(m,2H),7.37–7.29(m,3H),7.20(d,J＝7.8Hz,1H),6.12(d,J＝1.8Hz,1H),4.47(d,J＝5.6Hz,2H),3.80(s,3H),2.36(s,3H),1.95(s,3H)。
MS(ESI+)m/z 399.3[M+H]+。
Comparative example 10: n- ((1, 3-dimethyl-1H-pyrazol-4-yl) methyl) -4-methyl-3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000411
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1, 3-dimethyl-1H-pyrazol-4-yl) methylamine (0.063g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 10:1) to give the product as a white foamy solid (133mg, 65% yield).
1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.62(t,J＝5.5Hz,1H),7.62(d,J＝1.7Hz,1H),7.57(dd,J＝7.9,1.8Hz,1H),7.51(d,J＝8.3Hz,2H),7.46(s,1H),7.33(d,J＝8.0Hz,2H),7.17(d,J＝8.0Hz,1H),4.19(d,J＝5.5Hz,2H),3.70(s,3H),2.36(s,3H),2.11(s,3H),1.93(s,3H)。
MS(ESI+)m/z 412.8[M+H]+。
Comparative example 11: 4-methyl-3- ((4-methylphenyl) sulfonylamino) -N- ((1,3, 5-trimethyl-1H-pyrazol-4-yl) methyl) benzamide
Figure BDA0001894123760000412
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1,3, 5-trimethyl-1H-pyrazol-4-yl) methylamine (0.070g,0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (109mg, 51% yield).
1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.50(t,J＝5.4Hz,1H),7.59(d,J＝1.6Hz,1H),7.55(dd,J＝7.9,1.7Hz,1H),7.51(d,J＝8.2Hz,2H),7.33(d,J＝8.1Hz,2H),7.16(d,J＝8.0Hz,1H),4.15(d,J＝5.3Hz,2H),3.60(s,3H),2.36(s,3H),2.20(s,3H),2.09(s,3H),1.93(s,3H)。
MS(ESI+)m/z 426.8[M+H]+。
Comparative example 12: n- ((1, 5-dimethyl-1H-pyrazol-4-yl) methyl) -4-methyl-3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000421
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), (1, 5-dimethyl-1H-pyrazol-4-yl) methylamine (0.063g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (115mg, 56% yield).
1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.64(t,J＝5.6Hz,1H),7.60(d,J＝1.7Hz,1H),7.56(dd,J＝7.9,1.8Hz,1H),7.51(d,J＝8.3Hz,2H),7.33(d,J＝8.2Hz,2H),7.24(s,1H),7.17(d,J＝8.0Hz,1H),4.19(d,J＝5.6Hz,2H),3.68(s,3H),2.36(s,3H),2.22(s,3H),1.93(s,3H).
MS(ESI+)m/z 412.9[M+H]+.
Comparative example 13: 4-methyl-3- ((4-methylphenyl) sulfonylamino) -N- (pyrazolo [1,5-a ] pyridin-3-ylmethyl) benzamide
Figure BDA0001894123760000422
A reaction mixture of 4-methyl-3- ((4-methylphenyl) sulfonamido) benzoic acid (0.153g, 0.5mmol), pyrazolo [1,5-a ] pyridin-3-ylmethylamine (0.110g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (30 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (89mg, 41% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.87(t,J＝5.7Hz,1H),8.63(d,J＝7.0Hz,1H),7.94(s,1H),7.79(d,J＝9.0Hz,1H),7.63(d,J＝1.8Hz,1H),7.57(dd,J＝7.9,1.8Hz,1H),7.51(d,J＝8.3Hz,2H),7.32(d,J＝8.2Hz,2H),7.25–7.18(m,1H),7.17(d,J＝8.0Hz,1H),6.86(t,J＝6.8Hz,1H),4.57(d,J＝5.7Hz,2H),2.34(s,3H),1.92(s,3H)。
MS(ESI+)m/z 434.8[M+H]+。
Comparative example 14: 4-methyl-N- ((1-methyl-1H-imidazol-4-yl) methyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000431
A reaction mixture of 3- ((4-methylphenyl) sulfonamido) -4-methylbenzoic acid (0.153g, 0.5mmol), (1-methyl-1H-imidazol-4-yl) methylamine (0.056g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (94mg, 47% yield).
1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),8.70(t,J＝5.6Hz,1H),7.63(d,J＝1.7Hz,1H),7.60(dd,J＝7.9,1.8Hz,1H),7.52(d,J＝8.3Hz,2H),7.48(s,1H),7.34(d,J＝8.0Hz,2H),7.17(d,J＝7.9Hz,1H),6.92(s,1H),4.27(d,J＝5.6Hz,2H),3.60(s,3H),2.36(s,3H),1.94(s,3H)。
MS(ESI+)m/z 399.2[M+H]+。
Comparative example 15: 4-methyl-N- (2- (1-methyl-1H-pyrazol-4-yl) ethyl) -3- ((4-methylphenyl) sulfonylamino) benzamide
Figure BDA0001894123760000432
A reaction mixture of 3- ((4-methylphenyl) sulfonamido) -4-methylbenzoic acid (0.153g, 0.5mmol), 2- (1-methyl-1H-imidazol-4-yl) ethan-1-amine hydrochloride (0.063g, 0.5mmol), HATU (0.228g, 0.6mmol) and triethylamine (0.152g, 1.5mmol) in DCM (10mL) was stirred overnight. Water (30mL) was added and the resulting mixture was extracted with DCM (20 mL. times.3). The combined organic layers were washed with water (30 mL. times.2) and brine (30mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative thin layer chromatography (silica gel, DCM/MeOH ═ 15:1) to give the product as a white solid (141mg, 68% yield).
1H NMR(400MHz,DMSO-d6)δ9.62(s,1H),8.47(t,J＝5.6Hz,1H),7.62(d,J＝1.7Hz,1H),7.58–7.49(m,4H),7.34(d,J＝8.1Hz,2H),7.26(s,1H),7.19(d,J＝8.0Hz,1H),3.77(s,3H),3.39–3.33(m,2H),2.63(t,J＝7.4Hz,2H),2.36(s,3H),1.93(s,3H)。
MS(ESI+)m/z 413.2[M+H]+。
Evaluation of pharmacological Activity
Experimental example 1: determination of tumor cell survival by MTT method
Digesting DU145 cells (human prostate cancer cells) in logarithmic growth phase with 0.25% pancreatin-EDTA to prepare single cell suspension, inoculating 1500 cells/well/100 μ L into 96-well plate overnight, adding fresh culture medium containing different concentrations of test compound and corresponding solvent control, and adding 100 μ L (DMSO final concentration) per well<0.1 percent) of the total serum, continuously culturing for 72 hours at 37 ℃, adding 20 mu L of freshly prepared PBS solution containing 5mg/mL MTT into each hole, continuously culturing for 4 hours, removing supernatant, adding 180 mu L of DMSO into each hole to dissolve MTT formazan precipitates, oscillating and uniformly mixing by a micro oscillator, measuring an optical density value (OD) under the condition of a detection wavelength of 570nm, taking tumor cells treated by the DMSO as a control group, calculating the inhibition rate of the compound to be detected on the growth of the tumor cells by the following formula, and calculating IC according to an intermediate effect equation50：
Inhibition (%) - (control mean OD value-dosing mean OD value)/control mean OD value × 100%
The results are shown in Table 1. The experimental results show that all examples show strong antiproliferative activity on human prostate cancer DU145, and IC thereof50The values are all less than 0.12 mu M; all comparative examples showed a weak antiproliferative activity on human prostate cancer DU145, the IC of which50The values are all greater than 1.0. mu.M.
Table 1: antiproliferative activity on human prostate cancer cell DU145
Figure BDA0001894123760000441
Figure BDA0001894123760000451
Figure BDA0001894123760000461
Figure BDA0001894123760000471
Figure BDA0001894123760000481
Experimental example 2: detection of expression of p-STAT3 by immunoblotting
Control and varying concentrations of DU145 cells treated for 16h in examples 1-7, 12, 13 and 15 were collected, washed 2 times with pre-chilled PBS, and appropriate amounts of RIPA lysate (50mM Tris-HCI, 1mM EDTA, 1% Triton X-100, 150mM NaCl, 0.1% SDS, 1mM NaF, Na3VO4Protease inhibitor, pH 7.4) is cracked on ice for 1 hour, then centrifuged for 20min at 12,000rpm at4 ℃, the supernatant is collected, quantified and boiled for denaturation, the same amount of protein is taken for 10% SDS-PAGE electrophoresis, the primary antibody is incubated overnight at4 ℃ by using specific antibodies of p-STAT3(Tyr705) and STAT3, the secondary antibody marked by corresponding HRP is incubated for 2 hours at room temperature, washed, ECL chemiluminescence substrate reaction solution is added, developed in a gel imaging system, and images are stored, wherein β -actin is used as an internal reference.
The results are shown in FIG. 1.
Experimental example 3: efficacy study of human prostate cancer DU145 xenograft model in nude mice
Collecting human prostate cancer DU145 tumor cells under aseptic condition, adjusting cell density to 1 × 10 with sterilized normal saline6And (2) inoculating 0.2mL of the cells per liter to the axillary back subcutaneous part of the nude mouse, taking out the cells under aseptic conditions when the tumor grows to the size of 1cm in diameter, cutting the cells into tumor blocks with the size of 1mm multiplied by 1mm, and evenly inoculating the tumor blocks to the axillary back subcutaneous part of the nude mouse. After two weeks, the tumor grows to 100-300 mm3Thereafter, animals were randomized and dosing was initiated (day 0). Test compound portThe medicine is taken orally. Body weights were weighed twice weekly and tumor lengths and widths were measured with a vernier caliper. After the experiment, the nude mice were dislocated and sacrificed, the tumor tissue was peeled off, weighed and photographed. Finally, the tumor inhibition rate is calculated, and the anti-tumor effect strength is evaluated according to the tumor inhibition rate, and the results are shown in tables 3 and 4 and figures 2 and 3.
Tumor volume was calculated according to the following formula:
tumor volume ═ a × b2) And/2, a and b represent tumor body length and width, respectively.
The percent tumor growth inhibition was calculated according to the following formula: tumor growth inhibition (%) - (1-T/C) × 100, T is the tumor volume of the test compound group, and C is the tumor volume of the solvent control group.
The results are shown in Table 2 and FIG. 2. The experimental results show that example 1 shows significant antitumor activity in the human prostate cancer DU145 xenograft model in nude mice at doses of 10mg/kg and 20 mg/kg.
TABLE 2 growth inhibition in EXAMPLE 1 human prostate cancer DU145 xenograft model in nude mice
Figure BDA0001894123760000491
***p<0.001
Summary of pharmacological activity:
all examples show strong in vitro antiproliferative activity on human prostate cancer DU145 cells, IC50The values are all less than 0.12. mu.M. In immunoblotting experiments, examples 1-7, 12, 13 and 15 showed significant inhibitory activity on STAT3 phosphorylation of human prostate cancer DU 145. In vivo pharmacodynamic studies showed that example 1 showed significant growth inhibition in the human prostate cancer DU145 xenograft model in nude mice.

Claims (12)

1. A compound of formula (I), a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof:
Figure FDA0001894123750000011
wherein
R1Is selected from C1-4Alkyl radical, C3-6A cycloalkyl group;
R2selected from: phenyl, wherein the phenyl is substituted at the para-position with Ra; 5-10 membered heteroaryl, wherein the 5-10 membered heteroaryl is substituted with Ra; or with C4-6Cyclo-olefins or C4-6Heterocyclic olefin fused phenyl;
ra is selected from: chloro, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy or ethoxy;
R3and R4Each independently selected from hydrogen, halogen, methyl, ethyl or methoxy.
2. The compound, stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof according to claim 1,
R1selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl or cyclobutyl.
3. The compound, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R is2Selected from:
Figure FDA0001894123750000012
ra is selected from the group consisting of: chlorine, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy or ethoxy.
4. A compound, stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof according to claim 3, wherein said R2Selected from:
Figure FDA0001894123750000013
ra is selected from the group consisting of: methyl, ethyl, cyclopropyl, methoxy or ethoxy.
5. The compound, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R is2Selected from:
Figure FDA0001894123750000014
6. the compound, stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein R is3And R4Each independently selected from hydrogen, chlorine, fluorine, methyl, ethyl, and R3And R4At least one is hydrogen.
7. The compound, stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof according to claim 6, wherein said R is3Is methyl, R4Is hydrogen; or said R is3Is hydrogen, R4Is chlorine or fluorine.
8. The compound of claim 1, a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
Figure FDA0001894123750000021
9. a pharmaceutical composition comprising at least one compound according to any one of claims 1 to 8, a stereoisomer, a geometric isomer, a tautomer or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier and/or excipient.
10. The pharmaceutical composition of claim 9, wherein the pharmaceutical composition further comprises a pharmaceutically active ingredient other than the compound, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof.
11. Use of a compound according to any one of claims 1 to 8, a stereoisomer, a geometric isomer, a tautomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of claims 9 or 10, for the preparation of a medicament for the prophylaxis and/or treatment of STAT 3-mediated diseases.
12. The use according to claim 11, wherein said STAT 3-mediated disease comprises a tumor, an autoimmune disease, a renal disease, a cardiovascular disease, an inflammatory disease, a metabolic/endocrine dysfunction, or a neurological disease.
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CN108239075A (en) * 2016-12-26 2018-07-03 中国医学科学院药物研究所 Quinazoline compounds and preparation method thereof, purposes and pharmaceutical composition

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