CN115433107A - S1PR2 antagonist and application thereof in preparation of drugs for treating lung diseases - Google Patents

S1PR2 antagonist and application thereof in preparation of drugs for treating lung diseases Download PDF

Info

Publication number
CN115433107A
CN115433107A CN202211135871.5A CN202211135871A CN115433107A CN 115433107 A CN115433107 A CN 115433107A CN 202211135871 A CN202211135871 A CN 202211135871A CN 115433107 A CN115433107 A CN 115433107A
Authority
CN
China
Prior art keywords
alkyl
general formula
rings
group
compound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202211135871.5A
Other languages
Chinese (zh)
Other versions
CN115433107B (en
Inventor
万升标
佟晓永
曲显俊
赵万洲
罗栋栋
陈辉
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Ogpharmaceutical Technology Service Co ltd
Original Assignee
Nanjing Ogpharmaceutical Technology Service Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Ogpharmaceutical Technology Service Co ltd filed Critical Nanjing Ogpharmaceutical Technology Service Co ltd
Priority to CN202211135871.5A priority Critical patent/CN115433107B/en
Publication of CN115433107A publication Critical patent/CN115433107A/en
Application granted granted Critical
Publication of CN115433107B publication Critical patent/CN115433107B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C275/28—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C275/32—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by singly-bound oxygen atoms
    • C07C275/34—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by singly-bound oxygen atoms having nitrogen atoms of urea groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring
    • C07C275/36—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by singly-bound oxygen atoms having nitrogen atoms of urea groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring with at least one of the oxygen atoms further bound to a carbon atom of a six-membered aromatic ring, e.g. N-aryloxyphenylureas
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/62—Oxygen or sulfur atoms
    • C07D213/63—One oxygen atom
    • C07D213/68—One oxygen atom attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C07D213/75—Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/20—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carbonic acid, or sulfur or nitrogen analogues thereof
    • C07D295/215—Radicals derived from nitrogen analogues of carbonic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/22—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with hetero atoms directly attached to ring nitrogen atoms
    • C07D295/26—Sulfur atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2603/00—Systems containing at least three condensed rings
    • C07C2603/56—Ring systems containing bridged rings
    • C07C2603/58—Ring systems containing bridged rings containing three rings
    • C07C2603/70—Ring systems containing bridged rings containing three rings containing only six-membered rings
    • C07C2603/74—Adamantanes

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Urology & Nephrology (AREA)
  • Vascular Medicine (AREA)
  • Pulmonology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses an S1PR2 antagonist and application thereof in preparing a medicament for treating lung diseases. The compounds shown in general formulas I, II, III and IV, or pharmaceutically acceptable salts, solvates, prodrugs, active metabolites, N-oxides, stereoisomers, tautomers or geometrical isomers thereof are used as S1PR2 antagonists. The general formula compound is used for preparing medicines for inhibiting the expression of MMP2, IL-5 and TGF-beta 1 and increasing the expression of BMPR 2; or in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder or condition that would benefit from inhibition of expression of MMP2, IL-5, TGF- β 1, or increased expression of BMPR 2; or in the preparation of medicine for preventing and/or treating pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia, chronic tracheitis and other pulmonary diseases.

Description

S1PR2 antagonist and application thereof in preparation of drugs for treating lung diseases
Technical Field
The invention belongs to the field of medicines, relates to a novel S1PR2 antagonist, a pharmaceutical composition and application thereof, and particularly relates to an S1PR2 antagonist and application thereof in preparation of medicines for treating pulmonary hypertension, pulmonary stenosis, chronic obstructive pneumonia, chronic tracheitis and other pulmonary related diseases.
Background
Sphingosine-1-type phosphate receptor S1PR2, also known as EDG-5, is located on chromosome 19, has a total length of 9899nt, consists of 2 exons in total, encodes a protein consisting of 353 amino acid residues, has a molecular weight of 38kDa, and is involved in intracellular signal transduction, proliferation, differentiation, inflammation and immune response. The structure of the S1PR2 protein mainly comprises three parts: an N-terminal helix located outside the cell membrane, an alpha-helix region located inside the membrane, and a C-terminal helix region located inside the cell membrane that binds to G-proteins. S1PR2 plays an important role in cell survival, angiogenesis, vasodilation, inflammation, liver metabolism, immunity, muscle function, neuronal function, renal function, cancer, and the like (2016, heringdorf, etc.).
There is increasing evidence that S1P and S1PR signaling play a role in the development of pulmonary disease. Through research, the S1PR2 is found to be closely related to the expression of proteins such as bone morphogenetic protein receptor (BMPR 2), matrix metalloproteinase 2 (MMP 2), interleukin (IL-5), transforming growth factor-beta 1 (TGF-beta 1) and the like, and the proteins are used as markers and play an important role in pulmonary diseases such as pulmonary hypertension, pulmonary stenosis, chronic obstructive pneumonia, chronic tracheitis and the like. Antagonizing S1PR2 can effectively reduce the expression of MMP2, IL-5 and TGF-beta 1, promote the expression of BMPR2, and is beneficial to the treatment of pulmonary diseases such as pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia, chronic tracheitis and the like.
BMPR2 was originally determined to play a role in regulating the growth and maturation (differentiation) of bone and cartilage. BMPR2 was later found to play a broader role in regulating the growth and differentiation of many types of cells. BMPR2 inhibits growth factor-stimulated smooth muscle cell proliferation (2010, rabinovitch et al). Researchers have identified that more than 350 BMPR2 gene mutations are associated with pulmonary hypertension. The loss of BMPR2 results in the proliferation of pulmonary artery smooth muscle cells in response to TGF- β 1 and the bone morphogenic proteins themselves that normally inhibit its proliferation. Deletion of BMPR2 may also induce susceptibility to apoptosis.
MMP2 is one of members of the matrix metalloproteinase gene family (MMPs), MMP2 protein encoded by the gene mainly degrades collagen, MMP2 is widely involved in various physiological functions such as vascular remodeling, angiogenesis, tissue repair, tumor invasion, inflammation and atherosclerotic plaque rupture (2015, br a, etc.).
IL-5 is a major factor in the growth and differentiation of activated T and B cells, affecting the differentiation, activation, survival and proliferation of eosinophils, and is widely involved in disease progression. The use of anti-IL-5 antibody therapy has potential therapeutic effects in patients with asthma, atopic dermatitis, nasal polyposis, hypereosinophilic syndrome, eosinophilic esophagitis, and Churg-Strauss syndrome (2014, kutikhin et al).
Currently available diaryloxyphenyl S1PR2 antagonists are mainly used for treating fibrosis and PET-CT imaging (org. Biomol. Chem.,2015,13,7928, J.Med.chem.2021,64, 6037-6058), and no relevant reports on treating pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia, chronic tracheitis and other related diseases are found.
Disclosure of Invention
In view of the above problems, the present invention aims to provide a class of compounds which are structurally different from those reported in the literature and are used as S1PR2 antagonists, and applications of the compounds in preparing medicines for treating pulmonary diseases such as pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia and chronic tracheitis.
In a first aspect, the present invention provides compounds of formula I, II, III, IV, or pharmaceutically acceptable salts, solvates, prodrugs, active metabolites, N-oxides, stereoisomers, tautomers, or geometric isomers thereof, as S1PR2 antagonists:
Figure BDA0003852045040000021
in the general formula I, A represents aromatic ring or aromatic heterocyclic ring, including but not limited to pyridine ring, benzene ring, thiophene ring, thiazole ring and the like;
in the general formula I, R 1 Represents alkyl, aryl, alkanoyl, arylacyl, arylsulfonyl, alkylsulfonyl, alkyl groups containing a six-membered ring; wherein the alkyl in the alkyl, the alkyl acyl and the alkyl sulfonyl is the alkyl with 1-10 carbons;
x in the general formula I represents oxygen or sulfur;
in the general formula I, R 2 Represents fluorine, chlorine, bromine or iodine;
in the general formula I, R 3 Represents an alkoxy group, an alkylamide group, an ester group, a sulfonamide group, an acyl group or a sulfonyl group; wherein the alkyl in the alkoxy and the alkyl amide is the alkyl with 1-10 carbons;
in the general formula II, A represents alkylamino, alkylamino containing three-membered ring, four-membered ring, five-membered ring, six-membered ring, seven-membered ring, adamantyl, morpholine, piperazine or piperidine;
r in the general formula II 1 Represents alkyl, aryl, alkanoyl, arylacyl, arylsulfonyl, alkylsulfonyl or alkyl containing a six-membered ring; wherein the alkyl in the alkyl, the alkyl acyl and the alkyl sulfonyl is the alkyl with 1-10 carbons;
x in the general formula II represents oxygen or sulfur;
r in the general formula II 2 Represents fluorine, chlorine, bromine or iodine;
r in the general formula II 3 Represents an alkyl group of 1 to 10 carbons;
in the general formula III, A represents aromatic ring or aromatic heterocyclic ring, including but not limited to pyridine ring, benzene ring, thiophene ring, thiazole ring and the like;
in the general formula III R 1 Represents alkyl, aryl, alkanoyl, arylacyl, arylsulfonyl, alkylsulfonyl or alkyl containing a six-membered ring; wherein the alkyl in the alkyl, the alkyl acyl and the alkyl sulfonyl is the alkyl with 1-10 carbons;
x in the general formula III represents oxygen or sulfur;
in the general formula III R 2 Represents fluorine, chlorine, bromine or iodine;
in the general formula III R 3 Represents an alkoxy group, an alkylamide group, an ester group, a sulfonamide group, an acyl group or a sulfonyl group; wherein the alkyl group in the alkyl group, the alkylamide group, the alkylacyl group and the alkylsulfonyl group is 1-Alkyl of 10 carbons;
in the formula IV, A represents aromatic ring or aromatic heterocyclic ring, including but not limited to pyridine ring, benzene ring, thiophene ring, thiazole ring and the like;
in the general formula IV, B represents a pyridine ring or a benzene ring;
r in the general formula IV 1 Represents fluorine, chlorine, bromine, iodine, alkyl, alkoxy or acyl;
x in formula IV represents oxygen or sulfur;
in the general formula IV, y represents 1 or 2;
r in the general formula IV 2 Represents fluorine, chlorine, bromine or iodine;
r in the general formula IV 3 Represents an alkoxy group, an alkylamide group, an alkyl ester group, an alkylsulfonamide group, an alkanoyl group or an alkylsulfonyl group; wherein the alkyl in the alkoxy, alkyl amide, alkyl ester, alkyl sulfonamide, alkyl acyl or alkyl sulfonyl is 1-10 carbon alkyl.
In a second aspect, the present invention provides the use of a compound, or a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, as an antagonist of S1PR 2:
Figure BDA0003852045040000031
Figure BDA0003852045040000041
Figure BDA0003852045040000051
in a third aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the first aspect, a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, as described above, and a pharmaceutically acceptable dressing; optionally, the pharmaceutical composition further comprises one or more additional therapeutic agents.
In a fourth aspect, the present invention provides a compound according to the first aspect above, a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, or a pharmaceutical composition according to the third aspect above for use as an antagonist of S1PR 2.
In a fifth aspect, the present invention provides the use of a compound of the first aspect above, a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer or geometric isomer thereof, or a pharmaceutical composition of the third aspect above, or a combination thereof with one or more other therapeutic agents, in the manufacture of a medicament for the inhibition of the expression of MMP2, IL-5, TGF- β 1, the increase in the expression of BMPR 2; or in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder or condition that would benefit from inhibition of expression of MMP2, IL-5, TGF- β 1, or increased expression of BMPR 2; or in preparing medicine for preventing and/or treating pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia, chronic tracheitis, etc.
In a sixth aspect, the present invention provides a method for inhibiting the expression of MMP2, IL-5, TGF- β 1, promoting the expression of BMPR2 comprising administering to said cell an effective amount of a compound of the first aspect described above, a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer or geometric isomer thereof, or a pharmaceutical composition of the third aspect described above, or a combination thereof with one or more other therapeutic agents, or an antagonist of S1PR2 of the sixth aspect described above, wherein said method is performed in vitro.
In a seventh aspect, the present invention provides a method for preventing and/or treating pulmonary diseases such as pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pulmonary disease, chronic tracheitis, etc., comprising administering to the cell an effective amount of a compound of the first aspect, a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer or geometric isomer thereof, or a pharmaceutical composition of the third aspect, or a combination thereof with one or more other therapeutic agents.
The invention discovers for the first time that an S1PR2 antagonist can be used for inhibiting the expression of MMP2, IL-5 and TGF-beta 1 and promoting the expression of BMPR2 by inhibiting S1PR2, and BMPR2, MMP2, TGF-beta 1 and IL-5 are taken as disease markers and are closely related to the occurrence and development of diseases such as pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia and chronic tracheitis, and the provided general formula compound has potential application in treating pulmonary related diseases such as pulmonary hypertension, pulmonary artery stenosis, chronic obstructive pneumonia and chronic tracheitis related to BMPR2, MMP2, IL-5 and TGF-beta 1 as the S1PR2 antagonist.
Drawings
FIG. 1 is a Western blot experiment result chart showing that the compound synthesized by the present invention inhibits the expression of MMP2, IL-5 and TGF-beta 1 in Pulmonary Artery Smooth Muscle Cells (PASMCs) isolated from SKI mice and promotes the expression of BMPR 2.
FIG. 2 is a graph showing a comparison of the cytotoxic activity of the synthesized compounds of the present invention in isolated Pulmonary Artery Smooth Muscle Cells (PASMCs) from SKI mice.
Detailed Description
The following describes embodiments of the present invention in further detail with reference to specific examples. The following examples are for illustrative purposes only and are not intended to limit the scope of the present application.
Example 1:
the synthetic routes for compounds 29 and 37 are shown below, and compounds of formula I, II, III, IV can all be prepared by reference to the synthetic routes for compounds 29 and 37:
(1) Adding the compound T-1, CDI, methylamine hydrochloride, potassium carbonate and anhydrous magnesium sulfate into tetrahydrofuran, reacting overnight at 60 ℃, and washing and performing column chromatography on reaction liquid to obtain a compound T-2;
(2) Adding the compound T-2 and boron tribromide into dry dichloromethane, reacting for three hours at room temperature, washing reaction liquid and performing column chromatography to obtain a compound T-3;
(3) Adding the compound T-3, a nitro compound and cesium carbonate into dry N, N-diethylformamide, reacting overnight at 65 ℃, washing reaction liquid and performing column chromatography to obtain a compound T-4;
(4) Adding the compound T-4 and palladium carbon into ethyl acetate, adding a hydrogen balloon, ventilating by a water pump for three times, reacting for three hours at room temperature, filtering and concentrating to obtain a colloid; dissolving the colloid in ethyl acetate, adding 2, 2-ethyl trichloroformate and sodium bicarbonate, reacting at room temperature for two hours, washing the reaction solution with water and performing column chromatography to obtain a compound T-5;
(5) Adding a compound T-5, 1-Boc piperazine and diisopropylethylamine into N, N-diethylformamide, reacting overnight at 100 ℃, and washing and performing column chromatography on reaction liquid to obtain a compound T-6;
(6) Adding the compound T-6 and trifluoroacetic acid into tetrahydrofuran, reacting for three hours at room temperature, and concentrating to obtain a compound T-7;
(7) Adding the compound T-7, bromomethylcyclohexane and potassium carbonate into acetonitrile, reacting at 60 ℃ overnight, and washing reaction liquid with water and carrying out column chromatography to obtain a compound 29;
(7) Adding the compound T-7, 3-methoxyphenylacetic acid, DMAP and EDCI into dichloromethane, reacting overnight at room temperature, washing the reaction solution with water and performing column chromatography to obtain a compound 37.
Figure BDA0003852045040000071
Example 2:
the following S1PR2 antagonists were prepared according to the synthetic route of example 1:
Figure BDA0003852045040000081
Figure BDA0003852045040000091
1-(3,5-Bis(4-fluorophenoxy)phenyl)-3-(2,6-dichloropyridin-4-yl)urea(9)
1 H NMR(400MHz,DMSO-d 6 )δ9.47(s,1H),9.33(s,1H),7.49(s,2H),7.26(t,J＝8.7Hz, 4H),7.20–7.11(m,4H),6.84(d,J＝2.2Hz,2H),6.27(t,J＝2.2Hz,1H). 13 C NMR(100MHz, DMSO-d 6 )δ160.27,159.44,157.88,152.14,152.12,151.84,151.42,150.03,141.62,121.97, 121.88,117.27,117.04,111.45,102.78,102.23,31.14.HRESIMS m/z 502.0538[M+H] + (calcd for C 24 H 16 Cl 2 F 2 N 3 O 3 502.0531).
4-(3-(3-(2,6-Dichloropyridin-4-yl)ureido)-5-(4-fluorophenoxy)phenoxy)-N-methylbenzamide (10)
1 H NMR(400MHz,DMSO-d 6 )δ9.43(d,J＝48.1Hz,2H),8.42(q,J＝4.4Hz,1H),7.95– 7.87(m,2H),7.51(s,2H),7.27(t,J＝8.7Hz,2H),7.22–7.11(m,4H),6.98(t,J＝2.0Hz,1H), 6.90(t,J＝2.0Hz,1H),6.38(t,J＝2.2Hz,1H),2.81(d,J＝4.5Hz,3H). 13 C NMR(101MHz, DMSO-d 6 )δ166.34,159.51,158.84,158.01,152.12,151.85,151.38,150.03,141.74,130.38, 129.72,121.98,121.89,118.62,117.28,117.05,111.46,104.10,103.59,103.46,26.71.HRESIMS m/z 541.0844[M+H] + (calcd for C 26 H 20 Cl 2 FN 4 O 4 541.0840).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(pyridin-4-yloxy)phenyl)piperazine-1-carboxami de(11)
1 H NMR(400MHz,CDCl 3 )δ7.59–7.51(m,3H),7.33(s,1H),7.05(qd,J＝9.4,7.7,3.7Hz, 5H),6.56–6.52(m,1H),6.44–6.38(m,2H),3.63–3.49(m,4H),2.45–2.35(m,4H),2.17(d,J ＝6.8Hz,2H),1.44–1.31(m,5H),0.85(t,J＝7.4Hz,6H). 13 C NMR(100MHz,CDCl 3 )δ179.04, 159.59,154.45,142.88,139.11,121.42,121.34,118.79,116.86,116.63,108.41,107.94,105.69, 62.39,62.26,44.21,37.56,29.71,23.97,23.93,10.76.HRESIMS m/z 493.2602[M+H] + (calcd for C 28 H 34 FN 4 O 3 493.2609).
Tert-butyl (1-((3-(4-fluorophenoxy)-5-(pyridin-4-yloxy)phenyl)carbamoyl)piperidin-4-yl)carbamate(12)
1 H NMR(400MHz,DMSO-d 6 )δ8.81(s,1H),7.95–7.90(m,2H),7.48(t,J＝2.0Hz,1H), 7.30–7.25(m,2H),7.19–7.13(m,3H),6.87(d,J＝7.9Hz,1H),6.83(t,J＝2.2Hz,1H),6.24– 6.20(m,2H),3.99(d,J＝13.8Hz,2H),3.45(s,1H),2.86(t,J＝11.7Hz,2H),1.72(d,J＝10.4Hz, 2H),1.38(s,9H),1.26(d,J＝11.8Hz,2H). 13 C NMR(100MHz,DMSO-d 6 )δ177.89,158.82, 155.28,154.58,144.33,143.91,140.01,121.90,121.82,118.40,117.34,117.10,108.08,107.00, 105.81,78.07,47.68,43.20,32.25,28.73.HRESIMS m/z 523.2351[M+H] + (calcd for C 28 H 32 FN 4 O 5 523.2351).
4-((2-Ethylbutyl)amino)-N-(3-(4-fluorophenoxy)-5-(pyridin-4-yloxy)phenyl)piperidine-1-car boxamide(13)
1 H NMR(400MHz,DMSO-d 6 )δ7.91(d,J＝7.7Hz,2H),7.27(dd,J＝17.2,8.3Hz,4H), 7.15(dd,J＝9.1,4.5Hz,2H),7.04(t,J＝1.9Hz,1H),6.72(t,J＝2.2Hz,1H),6.20(d,J＝7.8Hz, 2H),2.67(s,2H),2.07(d,J＝7.2Hz,2H),1.98(dq,J＝17.1,10.3,8.8Hz,5H),1.73(d,J＝12.5 Hz,2H),1.49–1.31(m,6H),0.82(t,J＝7.4Hz,7H).HRESIMS m/z 507.2758[M+H] + (calcd for C 29 H 36 FN 4 O 3 507.2766).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)piperazine -1-carboxamide(14)
1 H NMR(400MHz,DMSO-d 6 )δ8.62(s,1H),8.39(q,J＝4.5Hz,1H),7.87(d,J＝8.7Hz, 2H),7.25(t,J＝8.7Hz,2H),7.17–7.06(m,4H),7.04(t,J＝2.1Hz,1H),6.98(t,J＝2.1Hz,1H), 6.29(t,J＝2.2Hz,1H),3.36(s,4H),2.78(d,J＝4.5Hz,3H),2.28(t,J＝4.9Hz,4H),2.10(d,J＝ 7.2Hz,2H),1.38–1.17(m,5H),0.82(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ 166.34,159.16,159.06,157.55,154.80,152.46,143.81,130.09,129.66,121.79,121.71,118.47, 117.21,116.98,104.52,104.01,102.64,62.20,53.63,44.21,37.14,26.71,23.86,11.01.HRESIMS m/z 549.2877[M+H] + (calcd for C 31 H 38 FN 4 O 4 549.2872).
5-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)hexahydro pyrrolo[3,4-c]pyrrole-2(1H)-carboxamide(15)
1 H NMR(400MHz,DMSO-d 6 )δ8.42(q,J＝4.5Hz,2H),7.91–7.84(m,2H),7.29–7.22 (m,2H),7.16–7.02(m,6H),6.30(s,1H),3.81(s,5H),2.78(d,J＝4.5Hz,8H),1.59(s,7H),0.83 (t,J＝7.2Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ166.30,159.13,159.05,157.53,152.44, 130.07,129.67,121.78,121.69,118.44,117.22,116.99,104.44,103.93,102.71,39.65,26.71,10.52. HRESIMS m/z 575.3032[M+H] + (calcd for C 33 H 40 FN 4 O 4 575.3028).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)-1,4-diaze pane-1-carboxamide(16)
1 H NMR(400MHz,DMSO-d 6 )δ8.43–8.31(m,2H),7.91–7.84(m,2H),7.28–7.20(m, 2H),7.15–7.04(m,6H),6.28(t,J＝2.2Hz,1H),3.51–3.40(m,6H),2.78(d,J＝4.5Hz,3H), 2.57(t,J＝5.2Hz,2H),2.21(d,J＝6.5Hz,2H),1.71(t,J＝5.8Hz,2H),1.28–1.22(m,5H),0.79 (t,J＝7.3Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ166.33,159.28,158.86,157.31,155.01, 152.59,144.02,129.95,129.63,121.58,121.50,118.24,117.17,116.93,105.10,104.60,102.75, 65.49,60.77,56.14,55.15,30.48,28.11,26.70,23.79,11.08.HRESIMS m/z 563.3038[M+H] + (calcd for C 32 H 40 FN 4 O 4 563.3028).
Methyl 4-(3-(4-(2-ethylbutyl)piperazine-1-carboxamido)-5-(4-fluorophenoxy)phenoxy) benzoate(17)
1 H NMR(400MHz,CDCl 3 )δ7.98(d,J＝8.8Hz,2H),7.08–6.95(m,6H),6.85(dt,J＝9.7, 2.1Hz,2H),6.56(s,1H),3.89(s,3H),3.44(t,J＝5.0Hz,4H),2.39(t,J＝4.9Hz,4H),2.16(d,J＝ 6.9Hz,2H),1.42–1.30(m,5H),0.85(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ 166.69,161.22,159.54,157.17,154.35,152.06,141.73,131.78,124.84,121.20,121.11,117.73, 116.65,116.42,105.65,105.03,104.23,77.45,77.14,76.81,62.39,53.28,52.16,44.10,37.47, 30.40,29.81,23.98,10.82.HRESIMS m/z 550.2725[M+H] + (calcd for C 31 H 37 FN 3 O 5 550.2712).
4-(3-(4-(2-Ethylbutyl)piperazine-1-carboxamido)-5-(4-fluorophenoxy)phenoxy)benzoic acid (18)
1 H NMR(400MHz,DMSO-d 6 )δ12.86(s,1H),9.16(s,1H),8.00–7.92(m,2H),7.51(s,1H), 7.38(s,1H),7.27(d,J＝2.4Hz,1H),7.24(d,J＝2.4Hz,1H),7.16–7.06(m,6H),6.34(t,J＝2.3 Hz,1H),4.35–4.05(m,2H),3.38(s,4H),2.95(s,4H),1.73(p,J＝6.2Hz,1H),1.39(m,J＝28.0, 14.0,6.6Hz,4H),0.84(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ167.22,160.91, 159.14,156.94,154.62,152.38,143.67,132.19,126.09,121.88,121.79,118.27,117.33,117.10, 105.22,104.60,103.38,59.82,56.52,51.67,34.96,23.63,19.09,10.62.HRESIMS m/z 536.2567 [M+H] + (calcd for C 30 H 35 FN 3 O 5 536.2555).
N-(3-(4-(dimethylcarbamoyl)phenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-ethylbutyl)piperazin e-1-carboxamide(19)
1 H NMR(400MHz,DMSO-d 6 )δ8.57(s,1H),7.41–7.34(m,2H),7.17(t,J＝8.7Hz,2H), 7.10–7.03(m,2H),7.03–6.89(m,4H),6.20(t,J＝2.2Hz,1H),3.30(d,J＝5.3Hz,4H),2.88(s, 6H),2.21(s,3H),2.03(d,J＝6.9Hz,2H),1.37(s,1H),1.21(dt,J＝10.2,5.1Hz,4H),0.75(t,J＝ 7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ170.05,159.14,157.72,157.64,154.82,152.43, 131.96,129.76,121.92,121.83,118.68,117.28,117.04,104.40,103.85,102.47,44.24,38.58,23.84, 22.95,11.04.HRESIMS m/z 563.3033[M+H] + (calcd for C 32 H 40 FN 4 O 4 563.3028).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(prop-2-yn-1-ylcarbamoyl)phenoxy)phenyl)pi perazine-1-carboxamide(20)
1 H NMR(400MHz,DMSO-d 6 )δ8.90(t,J＝5.6Hz,1H),8.63(s,1H),7.90(d,J＝8.7Hz, 2H),7.25(t,J＝8.7Hz,2H),7.17–7.07(m,4H),7.01(dt,J＝18.8,2.0Hz,2H),6.30(t,J＝2.2 Hz,1H),4.05(dd,J＝5.6,2.5Hz,2H),3.38(s,4H),3.12(t,J＝2.5Hz,1H),2.28(t,J＝4.9Hz, 4H),2.10(d,J＝7.2Hz,2H),1.43(p,J＝6.2Hz,1H),1.32–1.23(m,4H),0.81(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ165.62,159.52,159.07,157.41,154.78,143.81,129.94,129.28, 121.79,121.70,118.46,117.21,116.97,104.59,104.07,102.71,81.84,73.27,62.19,56.50,53.62, 44.19,37.12,28.94,23.85,19.02,11.01.HRESIMS m/z 573.2885[M+H] + (calcd for C 33 H 38 FN 4 O 4 573.2872).
N-(3-(4-carbamoylphenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-ethylbutyl)piperazine-1-carbo xamide(21)
1 H NMR(400MHz,DMSO-d 6 )δ8.62(s,1H),7.99–7.84(m,3H),7.32(s,1H),7.29–7.19 (m,2H),7.13(ddd,J＝6.8,5.2,2.8Hz,2H),7.10–7.05(m,2H),7.02(t,J＝2.0Hz,1H),6.98(t,J ＝2.0Hz,1H),3.37(s,4H),2.28(t,J＝4.8Hz,4H),2.10(d,J＝7.2Hz,2H),1.49–1.38(m,1H), 1.29(ddt,J＝18.1,13.9,7.3Hz,4H),0.82(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ167.63,159.35,159.13,157.58,143.83,130.20,129.85,121.89,121.81,118.47,117.29,117.06, 104.45,103.94,102.67,62.22,56.55,53.67,44.23,37.11,23.85,19.10,11.06.HRESIMS m/z 535.2725[M+H] + (calcd for C 30 H 36 FN 4 O 4 535.2715).
4-(2-Ethylbutyl)-N-(3-(4-(ethylcarbamoyl)phenoxy)-5-(4-fluorophenoxy)phenyl)piperazine-1 -carboxamide(22)
1 H NMR(400MHz,DMSO-d 6 )δ8.43(t,J＝5.6Hz,1H),7.92–7.80(m,2H),7.25(t,J＝8.8 Hz,2H),7.16–7.06(m,4H),7.02(q,J＝1.9Hz,1H),6.97(q,J＝1.9Hz,1H),6.28(t,J＝2.2Hz, 1H),3.27(tdd,J＝7.2,5.6,1.5Hz,2H),2.28(t,J＝4.9Hz,4H),2.10(d,J＝7.2Hz,2H),1.45(s, 1H),1.36–1.27(m,4H),1.11(t,J＝7.2Hz,3H),0.82(t,J＝7.4Hz,6H).HRESIMS m/z 563.3031 [M+H] + (calcd for C 32 H 40 FN 4 O 4 563.3028).
N-(3-(4-(cyclohexylcarbamoyl)phenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-ethylbutyl)piperaz ine-1-carboxamide(23)
1 H NMR(400MHz,DMSO-d 6 )δ8.63(s,1H),8.17(d,J＝8.0Hz,1H),7.93–7.83(m,2H), 7.31–7.20(m,2H),7.16–7.05(m,4H),6.99(dt,J＝22.7,2.0Hz,2H),3.36(s,4H),2.28(s,4H), 2.19–2.01(m,2H),1.77(ddd,J＝33.2,8.5,4.1Hz,4H),1.61(d,J＝12.6Hz,1H),1.44(s,1H), 1.33–1.23(m,8H),1.18–1.06(m,1H),0.82(t,J＝7.4Hz,7H). 13 C NMR(100MHz,DMSO-d 6 ) δ165.02,159.03,158.93,157.73,154.78,143.74,129.89,121.77,121.69,118.55,117.21,116.98, 104.30,103.88,102.46,62.16,53.59,48.80,44.17,37.08,32.92,25.74,25.43,23.83,10.99. HRESIMS m/z 617.3503[M+H] + (calcd for C 36 H 46 FN 4 O 4 617.3498).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(morpholine-4-carbonyl)phenoxy)phenyl)pipe razine-1-carboxamide(24)
1 H NMR(400MHz,DMSO-d 6 )δ7.49–7.42(m,2H),7.30–7.20(m,2H),7.17–7.10(m, 2H),7.10–7.05(m,2H),7.03(t,J＝2.0Hz,1H),6.97(t,J＝2.0Hz,1H),3.55(m,8H),3.38(s, 4H),2.28(t,J＝4.8Hz,4H),2.10(d,J＝7.1Hz,2H),1.38–1.28(m,4H),0.82(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ169.10,159.15,157.93,130.97,129.89,121.93,121.84,118.76, 117.29,117.06,66.60,53.67,44.23,37.11,23.86,11.06.HRESIMS m/z 605.3146[M+H] + (calcd for C 34 H 42 FN 4 O 5 605.3134).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(4-methylpiperazine-1-carbonyl)phenoxy)phe nyl)piperazine-1-carboxamide(25)
1 H NMR(400MHz,DMSO-d 6 )δ7.46–7.38(m,2H),7.29–7.20(m,2H),7.16–7.10(m, 2H),7.10–7.05(m,2H),7.01(dq,J＝19.4,1.9Hz,2H),3.37(d,J＝4.9Hz,8H),2.39–2.24(m, 8H),2.20(s,3H),2.10(d,J＝7.2Hz,2H),1.44(p,J＝6.2Hz,1H),1.32–1.23(m,4H),0.82(t,J ＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ168.98,160.17,159.15,157.84,157.62,154.77, 152.43,143.74,131.35,129.74,121.93,121.84,118.75,117.29,117.06,104.41,103.81,102.54, 62.22,53.67,46.13,44.23,37.12,23.86,11.06.HRESIMS m/z 618.3445[M+H] + (calcd for C 35 H 45 FN 5 O 4 618.3450).
4-(2-Ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(piperidine-1-carbonyl)phenoxy)phenyl)piper azine-1-carboxamide(26)
1 H NMR(400MHz,DMSO-d 6 )δ7.45–7.36(m,2H),7.28–7.20(m,2H),7.14–6.97(m, 6H),3.38(s,8H),2.29(t,J＝4.9Hz,4H),2.10(d,J＝7.2Hz,2H),1.61(q,J＝4.8,3.6Hz,2H), 1.50(s,5H),1.36–1.25(m,4H),0.82(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ 167.63,159.36,159.14,157.58,154.82,152.44,143.83,130.20,129.84,121.90,121.81,118.47, 117.30,117.06,104.44,103.92,102.65,62.22,56.55,53.67,44.23,37.09,23.84,19.10,11.05. HRESIMS m/z 603.3340[M+H] + (calcd for C 35 H 44 FN 4 O 4 603.3341).
N-(3-(4-(azepane-1-carbonyl)phenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-ethylbutyl)piperazi ne-1-carboxamide(27)
1 H NMR(400MHz,DMSO-d 6 )δ8.63(s,1H),7.43–7.36(m,2H),7.29–7.20(m,2H),7.17 –7.09(m,2H),7.09–7.03(m,2H),7.00(dt,J＝18.6,2.0Hz,2H),6.25(t,J＝2.2Hz,1H),3.54(t, J＝5.8Hz,2H),3.36–3.30(m,6H),2.29(t,J＝5.0Hz,4H),2.11(d,J＝7.2Hz,2H),1.55(s,7H), 1.32–1.24(m,4H),0.82(t,J＝7.4Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ199.74,193.82, 170.23,159.13,157.78,157.28,154.85,152.42,151.45,143.82,132.94,129.03,121.93,121.84, 118.83,117.28,117.05,104.37,103.79,102.38,71.54,62.22,53.67,49.71,45.98,44.23,37.11, 29.33,27.78,27.36,26.32,23.86,19.09,11.06,9.84.HRESIMS m/z 617.3506[M+H] + (calcd for C 36 H 46 FN 4 O 4 617.3498).
Tert-butyl 4-((3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)carbamoyl)piperazine-1-carbox ylate(28)
1 H NMR(400MHz,DMSO-d 6 )δ8.69(s,1H),8.39(q,J＝4.4Hz,1H),7.90–7.82(m,2H), 7.29–7.21(m,2H),7.17–7.06(m,4H),7.01(t,J＝2.0Hz,1H),6.96(t,J＝2.0Hz,1H),6.31(t,J ＝2.3Hz,1H),3.36(dd,J＝7.2,3.4Hz,4H),3.30(d,J＝5.4Hz,4H),2.78(d,J＝4.5Hz,3H), 1.40(s,9H). 13 C NMR(100MHz,DMSO-d 6 )δ166.32,159.11,159.08,157.61,154.83,154.29, 143.63,130.14,129.66,121.84,121.76,118.53,117.23,116.99,104.48,103.96,102.73,79.57, 43.92,28.50,26.71.HRESIMS m/z 565.2473[M+H] + (calcd for C 30 H 34 FN 4 O 6 565.2457).
4-(Cyclohexylmethyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)piper azine-1-carboxamide(29)
1 H NMR(400MHz,DMSO-d 6 )δ8.63(s,1H),8.39(q,J＝4.5Hz,1H),7.90–7.84(m,2H), 7.29–7.19(m,2H),7.16–7.05(m,4H),7.00(dt,J＝20.3,2.0Hz,2H),6.29(t,J＝2.3Hz,1H), 3.36(s,3H),2.78(d,J＝4.5Hz,3H),2.27(s,4H),2.07(d,J＝6.5Hz,2H),1.80–1.39(m,6H), 1.28–1.05(m,4H),0.83(t,J＝11.3Hz,2H). 13 C NMR(100MHz,DMSO)δ166.33,160.11, 159.13,159.05,157.72,157.54,154.79,152.46,143.79,130.09,129.65,121.77,121.69,118.46, 117.20,116.97,104.52,104.00,102.63,65.24,53.54,44.15,34.66,31.69,26.83,26.71,25.97. HRESIMS m/z 561.2878[M+H] + (calcd for C 32 H 38 FN 4 O 4 561.2872).
4-(4-Chlorophenyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)piperaz ine-1-carboxamide(30)
1 H NMR(400MHz,DMSO-d 6 )δ8.77(s,1H),8.41(q,J＝4.4Hz,1H),7.96–7.82(m,2H), 7.30–7.21(m,4H),7.19–7.06(m,5H),7.03(t,J＝2.1Hz,1H),7.00–6.93(m,2H),6.33(t,J＝ 2.3Hz,1H),3.55(t,J＝5.0Hz,4H),3.13(t,J＝5.1Hz,4H),2.80(d,J＝4.5Hz,3H). 13 C NMR (100MHz,DMSO)δ166.36,160.13,159.14,159.10,157.74,157.59,154.87,152.45,152.42, 150.11,143.70,130.12,129.67,129.10,123.23,121.79,121.70,118.49,117.67,117.20,116.97, 104.58,104.06,102.75,48.53,43.89,26.71.HRESIMS m/z 575.1871[M+H] + (calcd for C 31 H 29 ClFN 4 O 4 575.1856).
4-(4-Fluorobenzyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)piperaz ine-1-carboxamide(31)
1 H NMR(400MHz,DMSO-d 6 )δ8.63(s,1H),8.39(q,J＝4.5Hz,1H),7.89–7.83(m,2H), 7.33(dd,J＝8.5,5.8Hz,2H),7.25(t,J＝8.7Hz,2H),7.17–7.11(m,4H),7.10–7.06(m,2H), 6.99(dt,J＝20.9,2.0Hz,2H),6.29(t,J＝2.3Hz,1H),3.46(s,2H),3.37(t,J＝5.0Hz,4H),2.78 (d,J＝4.5Hz,3H),2.32(t,J＝4.9Hz,4H). 13 C NMR(100MHz,DMSO)δ166.32,160.11,159.11, 159.06,157.73,157.56,154.81,152.44,143.76,134.50,131.22,131.14,130.10,129.65,121.80, 121.72,118.49,117.21,116.98,115.49,115.28,104.48,103.96,102.64,61.41,52.76,44.13,26.71. HRESIMS m/z 573.2308[M+H] + (calcd for C 32 H 31 F 2 N 4 O 4 573.2308).
N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)-4-(4-(trifluoromethyl)phen yl)piperazine-1-carboxamide(32)
1 H NMR(400MHz,DMSO-d 6 )δ8.77(s,1H),8.41(q,J＝4.4Hz,1H),7.94–7.84(m,2H), 7.52(d,J＝8.8Hz,2H),7.31–7.22(m,2H),7.19–7.05(m,7H),7.02(t,J＝2.0Hz,1H),6.32(t, J＝2.2Hz,1H),3.56(dd,J＝6.7,3.8Hz,4H),3.35–3.25(m,4H),2.79(d,J＝4.5Hz,3H). 13 C NMR(101MHz,DMSO)δ166.34,160.14,159.12,157.75,157.60,154.87,153.51,152.44,143.67, 130.12,129.67,126.77,126.65,126.61,124.08,121.81,121.72,118.50,117.21,116.98,114.82, 104.56,104.04,102.77,47.28,43.68,26.70.HRESIMS m/z 609.2133[M+H] + (calcd for C 32 H 29 F 4 N 4 O 4 609.2119).
N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)-4-(3-methoxyphenyl)piper azine-1-carboxamide(33)
1 H NMR(400MHz,DMSO-d 6 )δ8.71(s,1H),8.37(d,J＝4.9Hz,1H),7.94–7.76(m,2H), 7.27–7.17(m,2H),7.15–7.00(m,6H),6.97(d,J＝6.0Hz,1H),6.59–6.21(m,4H),3.67(s,3H), 3.48(s,4H),3.07(s,4H),2.75(s,3H). 13 C NMR(100MHz,DMSO-D 6 )δ166.37,160.69,160.19, 159.17,157.63,154.91,152.76,152.46,152.43,143.77,130.18,130.13,129.73,121.92,121.83, 118.56,117.30,117.07,108.93,105.03,104.51,103.97,102.87,102.75,102.48,56.56,55.40,48.79, 44.04,26.79,19.10.HRESIMS m/z 571.2365[M+H] + (calcd for C 32 H 32 FN 4 O 5 571.2351).
N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)-4-(4-methoxyphenyl)piper azine-1-carboxamide(34)
1 H NMR(400MHz,DMSO-d 6 )δ8.72(s,1H),8.38(d,J＝4.6Hz,1H),7.93–7.81(m,2H), 7.25(t,J＝8.8Hz,2H),7.17–7.11(m,2H),7.11–7.07(m,2H),7.03(dt,J＝20.9,2.0Hz,2H), 6.94–6.88(m,2H),6.85–6.79(m,2H),6.30(t,J＝2.2Hz,1H),3.68(s,3H),3.52(t,J＝5.1Hz, 4H),2.97(t,J＝5.0Hz,4H),2.78(d,J＝4.5Hz,3H). 13 C NMR(100MHz,DMSO)δ166.33, 160.13,159.12,159.09,157.74,157.58,154.86,153.74,152.44,145.69,143.73,130.11,129.66, 121.81,121.73,118.49,118.43,117.22,116.99,114.73,104.51,104.00,102.70,55.64,50.30,44.18, 26.71.HRESIMS m/z 571.2366[M+H] + (calcd for C 32 H 32 FN 4 O 5 571.2351).
4-(4-Fluorobenzoyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)pipera zine-1-carboxamide(35)
1 H NMR(400MHz,DMSO-d 6 )δ8.75(s,1H),8.40(q,J＝4.5Hz,1H),7.93–7.83(m,2H), 7.55–7.45(m,2H),7.33–7.21(m,4H),7.18–7.06(m,4H),7.00(dt,J＝22.3,2.0Hz,2H),6.32 (t,J＝2.3Hz,1H),3.45(s,8H),2.79(d,J＝4.5Hz,3H). 13 C NMR(100MHz,DMSO)δ168.77, 166.33,164.30,161.84,160.15,159.13,159.08,157.76,157.64,154.84,152.40,143.60,132.56, 132.53,130.16,130.08,129.67,121.85,121.77,118.54,117.22,116.99,115.98,115.77,104.49, 103.96,102.76,44.08,26.71.HRESIMS m/z 587.2117[M+H] + (calcd for C 32 H 29 F 2 N 4 O 5 587.2101).
N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)-4-tosylpiperazine-1-carbox amide(36)
1 H NMR(400MHz,DMSO-d 6 )δ8.74(s,1H),8.43(q,J＝4.5Hz,1H),7.95–7.86(m,2H), 7.65(d,J＝8.3Hz,2H),7.46(d,J＝8.1Hz,2H),7.25(t,J＝8.7Hz,2H),7.19–7.06(m,4H),6.98 (dt,J＝20.2,2.0Hz,2H),6.33(t,J＝2.2Hz,1H),3.51(t,J＝4.8Hz,4H),2.85(m,7H),2.41(s, 3H). 13 C NMR(101MHz,DMSO)δ166.37,160.13,159.12,159.06,157.75,157.63,154.53, 152.36,152.34,144.32,143.45,132.11,130.34,130.15,129.66,128.07,121.79,121.71,118.52, 117.16,116.93,104.43,103.91,102.76,46.26,43.51,26.71,21.46.HRESIMS m/z 619.2036 [M+H] + (calcd for C 32 H 32 FN 4 O 6 S 619.2021).
N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)-4-(2-(3-methoxyphenyl)ac etyl)piperazine-1-carboxamide(37)
1 H NMR(400MHz,DMSO-d 6 )δ8.71(s,1H),8.41(q,J＝4.5Hz,1H),7.89(d,J＝8.7Hz, 2H),7.24(dt,J＝11.5,8.4Hz,3H),7.18–7.07(m,4H),7.02(dt,J＝21.9,2.0Hz,2H),6.87–6.75 (m,3H),6.33(t,J＝2.2Hz,1H),3.73(d,J＝4.0Hz,5H),3.49(dd,J＝6.9,3.7Hz,4H),3.39– 3.26(m,4H),2.80(d,J＝4.5Hz,3H). 13 C NMR(101MHz,DMSO)δ169.44,166.36,160.14, 159.72,159.11,157.75,157.61,154.81,152.43,143.62,137.66,130.14,129.81,129.67,121.80, 121.72,121.59,118.50,117.20,116.97,115.13,112.28,104.54,104.02,102.78,55.38,55.36,45.71, 44.19,43.92,41.55,26.71.HRESIMS m/z 613.2473[M+H] + (calcd for C 34 H 34 FN 4 O 6 613.2457).
4-(2-Naphthoyl)-N-(3-(4-fluorophenoxy)-5-(4-(methylcarbamoyl)phenoxy)phenyl)piperazine -1-carboxamide(38)
1 H NMR(400MHz,DMSO-d 6 )δ8.81(s,1H),8.43(q,J＝4.5Hz,1H),8.09–7.84(m,6H), 7.64–7.50(m,3H),7.31–6.97(m,8H),6.35(s,1H),3.88–3.44(m,8H),2.81(d,J＝4.5Hz,3H). 13 C NMR(101MHz,DMSO)δ169.68,166.38,160.15,159.16,159.11,157.77,157.67,154.91, 152.42,152.40,143.64,133.63,133.51,132.70,130.17,129.69,128.83,128.54,128.16,127.62, 127.23,127.10,124.97,121.84,121.75,118.55,117.21,116.98,104.56,104.04,102.79,55.36, 44.21,26.72.HRESIMS m/z 619.2368[M+H] + (calcd for C 36 H 32 FN 4 O 5 619.2351).
4-(2-ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-methoxyphenoxy)phenyl)piperazine-1-carboxa mide(39)
1 H NMR(400MHz,DMSO-d 6 )δ8.58(s,1H),7.16(d,J＝9.8Hz,2H),7.04(d,J＝4.5Hz, 2H),6.98(s,1H)，6.96(s,1H),6.91(d,J＝6.9Hz,2H),6.89(d,J＝4.0Hz,2H),6.10(s,1H),3.70 (s,3H),3.33(t,J＝4.0Hz,4H),2.23(t,J＝5.0Hz,4H),2.04(d,J＝7.2Hz,2H),1.27–1.16(m, 5H),0.76(t,J＝7.4Hz,6H)； 13 C NMR(100MHz,DMSO-d 6 )δ159.77,158.74,156.25,155.79, 149.31,143.66,121.59,121.54,121.50,117.14,116.91,115.50,103.00,102.93,100.81,55.84, 36.65.
4-(4-chlorophenyl)-N-(3-(4-fluorophenoxy)-5-(4-methoxyphenoxy)phenyl)piperazine-1-carb oxamide(40)
1 H NMR(400MHz,DMSO-d 6 )δ8.67(s,1H),7.26(d,J＝4.0Hz,2H),7.23(d,J＝3.5Hz, 2H),7.11(d,J＝4.0Hz,2H),7.05(s,1H),7.02(s,1H),6.98(d,J＝4.0Hz,2H),6.96(d,J＝4.0 Hz,2H),6.89(d,J＝7.9,Hz,2H),6.16(s,J＝2.2Hz,1H),3.76(s,3H),3.51(t,J＝5.1Hz,4H), 3.11(t,J＝5.1Hz,4H). 13 C NMR(100MHz,DMSO-d 6 )δ159.89,158.88,152.44,149.19,129.15, 123.26,121.71,121.63,117.75,117.19,116.96,115.53,102.78,100.94,55.85,48.54,43.86,39.12.
N-(3-(4-(diethylcarbamoyl)phenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-ethylbutyl)piperazine- 1-carboxamide(41)
1 H NMR(400MHz,DMSO-d 6 )δ7.39(d,J＝8.0Hz,2H),7.25(d,J＝4.0Hz,2H),7.14(d,J ＝4.0Hz,2H),7.08(d,J＝4.0Hz,2H),7.03(s,1H),6.98(s,1H),6.25(s,1H),3.36(t,J＝8.0Hz, 4H),3.35(q,4H),2.25(t,J＝8.0Hz,4H),2.10(d,J＝8.0Hz,2H),1.31–1.23(m,5H),1.10(t,J ＝8.0Hz,6H),0.83(t,J＝8.0Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ169.92,159.07,157.74, 157.32,154.76,152.41,132.82,128.78,121.84,121.75,118.81,117.21,116.98,104.32,103.84, 102.35,62.20,53.63,44.20,39.38,37.12,23.85,11.01.
4-(2-ethylbutyl)-N-(3-(4-fluorophenoxy)-5-(4-(isopropylcarbamoyl)phenoxy)phenyl)piperazi ne-1-carboxamide(42) 1 H NMR(400MHz,DMSO-d 6 )δ8.09(s,1H),7.89(s,1H),7.27–7.21(d, J＝8.0Hz,2H),7.14–7.11(d,J＝4.0Hz,2H),7.09–7.07(d,J＝8.0Hz,2H),7.03(s,1H),6.97(s, 1H),6.59–6.57(d,J＝8.0Hz,2H),6.27(s,1H),4.40–4.36(m,1H),3.37(t,J＝8.0Hz,4H),2.28 (t,J＝8.0Hz,4H),2.10(d,J＝4.0Hz,2H),1.26–1.22(m,5H),1.16(d,J＝8.0Hz,6H),0.82(t,J ＝8.0Hz,6H). 13 C NMR(100MHz,DMSO-d 6 )δ158.99,157.72,154.43,149.77,129.85,121.77, 121.68,118.52,117.20,116.97,107.16,102.46,62.19,56.50,56.38,53.62,44.19,40.44,39.36, 39.03,37.13,23.84,22.79,22.77,18.98,18.93,10.99.
N-(3-(4-(cyclopropylcarbamoyl)phenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-ethylbutyl)pipera zine-1-carboxamide(43)
1 H NMR(400MHz,DMSO-d 6 )δ8.09(s,1H),7.86(s,1H),7.24(d,J＝8.0Hz,2H),7.14(d,J ＝8.0Hz,2H),7.08(d,J＝4.0Hz,2H),7.03(s,1H),6.97(s,1H),6.58(d,J＝4.0Hz,2H),6.28(s, 1H),3.36(t,J＝4.8Hz,4H),2.82(m,1H),2.28(t,J＝4.9Hz,4H),2.10(d,J＝7.2Hz,2H),1.31– 1.23(m,5H),0.82(t,J＝7.4Hz,6H),0.69(dt,J＝7.2,2.4Hz,2H),0.56(dt,J＝4.3,2.5Hz,2H). 13 C NMR(100MHz,DMSO-d 6 )δ167.22,160.12,159.15,159.06,157.74,157.60,154.42,152.43, 149.80,143.79,130.00,129.80,121.80,121.71,118.48,117.23,116.99,107.17,104.43,103.95, 102.55,62.19,53.62,44.19,39.05,37.12,23.84,23.49,23.37,11.01,6.23.
N-(3-(4-(((3R,5R)-adamantan-1-yl)carbamoyl)phenoxy)-5-(4-fluorophenoxy)phenyl)-4-(2-et hylbutyl)piperazine-1-carboxamide(44)
1 H NMR(400MHz,DMSO-d 6 )δ7.84(d,J＝4.0Hz 2H),7.27(d,J＝4.0Hz,2H),7.14(d,J＝ 4.0Hz,2H),7.07(d,J＝4.0Hz,2H),7.02(s,1H),6.94(s,1H),6.26(s,1H),3.36(t,J＝4.0Hz, 4H),2.28(t,J＝5.0Hz,4H),2.10(d,J＝7.2Hz,2H),2.06(d,J＝4.0Hz,8H),1.66-1.64(m,3H), 1.27–1.24(m,5H),1.06(dd,J＝7.2Hz,4H),0.81(t,J＝7.5Hz,6H). 13 C NMR(100MHz, DMSO-d 6 )δ159.03,158.74,157.84,129.97,121.78,121.70,118.50,117.23,117.00,104.26, 103.82,102.38,62.19,56.50,53.62,51.94,44.19,41.35,39.41,37.12,36.56,29.36,23.84,19.01, 18.95,11.01.
methyl4-(3-(4-fluorophenoxy)-5-(3-(4-methoxyphenyl)ureido)phenoxy)benzoate(45)
1 H NMR(400MHz,CDCl 3 )δ8.03(d,J＝8.5Hz,2H),7.38(d,J＝8.7Hz,2H),7.07–7.03(m, 5H),7.01-6.99(m,5H),6.53(s,1H),3.90(s,3H),3.83(s,3H). 13 C NMR(100MHz,CDCl 3 )δ 160.19,157.99,151.19,149.03,137.60,131.89,126.96,125.73,122.19,121.55,121.47,118.24, 116.91,116.67,114.88,106.38,102.31,101.76,55.60,52.13.
methyl4-(3-(3-(4-acetylphenyl)ureido)-5-(4-fluorophenoxy)phenoxy)benzoate(46)
1 H NMR(400MHz,CDCl 3 )δ8.05(d,J＝4.0Hz,2H),7.58(d,J＝5.5Hz,2H),7.51(s,1H), 7.44(s,1H),7.05-7.01(m,7H),6.82(s,1H),6.73(s,1H),3.90(s,3H),3.89(s,3H). 13 C NMR(101 MHz,CDCl 3 )δ160.12,131.97,131.84,121.46,121.28,118.20,117.88,117.02,116.78,116.56, 112.17,111.14,107.93,107.36,29.71.
4-cyclohexyl-N-(3-(4-(4-ethylpiperazine-1-carbonyl)phenoxy)-5-(4-fluorophenoxy)phenyl)pi perazine-1-carboxamide(47)
1 H NMR(400MHz,DMSO-d 6 )δ8.61(s,1H),8.09(d,J＝8.0Hz,1H),7.41(d,J＝8.0Hz, 2H),7.25(d,J＝8.0Hz,2H),7.09(d,J＝8.0Hz,1H),7.04(d,J＝8.0Hz,2H),6.04(s,2H),6.23 (d,1H),2.97-2.95(m,2H),2.94(t,J＝4Hz,4H),2.49-2.46(m,1H),2.41(t,J＝4Hz,4H), 2.34-2.31(m,J＝8.0Hz,8H),1.70-1.67(m,4H),1.24–1.09(m,6H),0.97(t,J＝8.0Hz,3H). 13 C NMR(100MHz,DMSO-d 6 )δ168.88,157.79,157.60,154.81,152.42,148.00,129.68,121.83, 121.75,118.72,117.22,116.99,107.23,104.48,103.91,102.46,63.13,51.95,48.87,44.56,40.54, 39.44,39.25,28.69,26.32,25.76,12.32.
methyl4-(3-(3-(4-butylphenyl)ureido)-5-(4-fluorophenoxy)phenoxy)benzoate(48)
1 H NMR(400MHz,DMSO-d 6 )δ7.94(d,J＝8.0Hz,2H),7.39(d,J＝8.0Hz,2H),7.33(d,J ＝8.0Hz,2H),7.26(d,J＝8.0Hz,2H),7.21-7.15(m,4H),6.92(s,2H),6.65(s,1H),3.80(s,3H), 2.58-2.56(m,2H),1.58-1.48(m,2H),1.27-1.25(m,2H),0.86(t,J＝12.0Hz,3H). 13 C NMR(100 MHz,DMSO-d 6 )δ166.02,160.42,159.99,157.70,150.26,144.35,138.65,132.15,129.52,128.16, 126.97,125.63,122.38,122.30,118.89,117.54,117.30,106.49,102.85,102.21,52.60,39.45, 34.90,33.42,22.17,14.20.
example 3: surface plasmon resonance experiment
The affinity of the compounds for the S1PR2 protein was determined using a GE biacore t200 (GE, USA) instrument. S1PR2 protein was immobilized on a CM5 chip (GE, USA) in 15000 RU by amine coupling. After activation with sterile buffer (EDC/NHS, 1/1) at a flow rate of 30. Mu.L/min for 15 minutes, the S1PR2 proteins immobilized on the chip were blocked with ethanolamine for 8 minutes. After dissolving the compound in DMSO, the compound was diluted to a certain concentration with purified water and injected, PBSP buffer solution (1% DMSO) was used as a mobile phase, and the compound was flowed over the chip surface at a flow rate of 30. Mu.L/min for 60 seconds under the condition of 25 ℃. The dissociation rate constants of the compounds were determined by continuing the assay for 60 seconds and kinetic parameters were calculated using biacore t200SPR software. Kinetic fitting model fitting function plot using BiacoreT200SPR evaluation software, using binding Rate constant (K) a ) And dissociation rate constant (K) d ) Calculating outEquilibrium dissociation constant (K) D ). The results are shown in Table 1.
TABLE 1 binding Activity of the Compounds of the invention with S1PR2
Figure BDA0003852045040000191
Figure BDA0003852045040000201
Note: k D And EC 50 The value is measured at least 3 times (n is more than or equal to 3), and the error is less than 20 percent.
As can be seen from the data in table 1, most compounds bind well to S1PR2, exhibiting nanomolar binding activity, suggesting that these compounds may be useful as antagonists of S1PR2 for the treatment of related diseases.
Example 4: experiment for inhibiting MMP2, IL-5 and TGF-beta 1 expression and promoting BMPR2 expression in S1PR2 antagonist
Pulmonary Artery Smooth Muscle Cells (PASMCs) isolated from SKI mice (solvent control group, group administered with compounds 11, 13, 14, 18, 19, 22, 25, 28, 29 of table 1 alone) were removed and subjected to cell scraping, protein extraction and quantification.
Extracting cell protein: firstly, washing cells by PBS, precooling each group of cells at 4 ℃, and preparing cell lysate: NP-40 (960. Mu.L) + PPI (40. Mu.L) + PMSF (40. Mu.L). PBS was discarded, and 100-200. Mu.L of lysate was added depending on cell density, and the mixture was left at 4 ℃ for 40min. The cells were scraped off with a cell scraper and collected in a 1.5mL centrifuge tube, 12000rpm,15min for centrifugation. The supernatant was collected in another 1.5mL centrifuge tube, and 90. Mu.L of G250 staining solution was added to a 96-well plate, and 10. Mu.L of protein standard solution and 10. Mu.L of sample were added to prepare a duplicate well. The microplate reader determines the cell concentration, adjusts the cell concentration of each group to be consistent, adds 5 Xloading buffer solution to make the concentration to be 1 Xand boils for 5min in a metal bath.
Western blot detection: and (3) preparing separation gel and concentrated gel, and soaking in an electrophoresis solution. An equal amount of 20. Mu.g protein sample was added to the loading wells and labeled with either tag or 1 Xloading buffer. After the protein drops in the separation gel, the 80V constant voltage electrophoresis is changed into 120V, and the constant voltage electrophoresis is continued. The PVDF membrane was used for 100V,90min transfer. The PVDF membrane was washed once in 5min for 2 times in 1 XTBST on a shaker. The PVDF membrane was blocked with blocking solution (1h) and DPD primary antibody was incubated overnight at 4 ℃. The PVDF membrane was washed with 1 × TBST on a shaker for 3 times, once for 10 min. The secondary antibody was incubated for 40min. The PVDF membrane was washed with 1 XTBST on a shaker for 10min once for 3 times. ECL luminescence solution was prepared, and uniformly sprayed on PVDF membrane, and exposed by ChemiDoc XRS and molecular imager (molecular imager). Calculating the gray scale ratio of the analysis imprinting of the Bio-Rad Quantity One, and calculating a formula: relative expression level of the protein of interest = (grey value of protein of interest-background grey value)/(grey value of internal reference protein-background grey value).
The results show that S1PR2 antagonists 11, 13, 14, 18, 19, 22, 25, 28, 29 can effectively inhibit MMP2, IL-5 expression, promote BMPR2 expression compared to internal controls (see fig. 1).
Example 5: cytotoxic Activity of S1PR2 antagonists in Pulmonary Arterial Smooth Muscle Cells (PASMCs) isolated from SKI mice
In vitro cytotoxic activity assay: the log phase growth of PASMC cells were collected, the cell suspension concentration was adjusted, 100. Mu.L was added to each well, and the test cells were plated to 1000-10000/well (marginal wells filled with sterile PBS). 5% of CO 2 Incubate at 37 ℃ until the cell monolayer is confluent at the bottom of the wells (96-well flat bottom plate) and add a concentration gradient of the S1PR2 antagonist. 3 concentration gradients were set, 100 μ L per well, for 3 replicates. 5% of CO 2 Incubation at 37 ℃ and observation under an inverted microscope after different incubation times.
The results showed that there was no significant difference in the cell morphology and number of the administered group from the control group, indicating that most of the compounds did not have significant cytotoxic activity (see fig. 2).
In conclusion, the compound prepared by the invention can regulate the expression levels of BMPR2, IL-5, TGF-beta 1 and MMP2 of pulmonary artery smooth muscle cells by targeting antagonism of S1PR2 activity. Since these proteins are markers of pulmonary diseases such as pulmonary hypertension, pulmonary stenosis, chronic obstructive pneumonia, and chronic tracheitis, S1PR2 antagonists that inhibit the expression of MMP2, IL-5, and TGF- β 1 and promote the expression of BMPR2 by targeting S1PR2 are suitable for the treatment of pulmonary diseases such as pulmonary hypertension, pulmonary stenosis, chronic obstructive pneumonia, and chronic tracheitis.
The invention has been described in detail with respect to a general description and specific embodiments thereof, but it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Accordingly, it is intended that all such modifications and alterations be included within the scope of this invention as defined in the appended claims.

Claims (9)

1. A compound of formula I, II, III, IV, or a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, as an S1PR2 antagonist:
Figure FDA0003852045030000011
wherein:
in the general formula I, A is selected from aromatic rings or aromatic heterocycles and comprises pyridine rings, benzene rings, thiophene rings or thiazole rings;
in the general formula I, R 1 Selected from the group consisting of alkyl, aryl, alkanoyl, arylacyl, arylsulfonyl, alkylsulfonyl, alkyl groups containing a six-membered ring; wherein the alkyl in the alkyl, the alkyl acyl and the alkyl sulfonyl is the alkyl with 1-10 carbons;
x in the general formula I is selected from oxygen or sulfur;
in the general formula I, R 2 Selected from fluorine, chlorine, bromine or iodine;
in the general formula I, R 3 Selected from alkoxy, alkylamide, ester, sulfonamide, acyl or sulfonyl; wherein the alkyl in the alkoxy and the alkyl amide is the alkyl with 1-10 carbons;
in the general formula II, A is selected from alkylamino, alkylamino containing three-membered ring, four-membered ring, five-membered ring, six-membered ring, seven-membered ring and adamantyl, morpholine, piperazine or piperidine;
r in the general formula II 1 Selected from alkyl, aryl, alkanoyl, arylacyl, arylsulfonyl, alkylsulfonyl or alkyl groups containing a six-membered ring; wherein the alkyl in the alkyl, the alkyl acyl and the alkyl sulfonyl is the alkyl with 1-10 carbons;
x in the general formula II is selected from oxygen or sulfur;
r in the general formula II 2 Selected from fluorine, chlorine, bromine or iodine;
r in the general formula II 3 Selected from alkyl of 1-10 carbons;
in the general formula III, A is selected from aromatic rings or aromatic heterocycles, including pyridine rings, benzene rings, thiophene rings or thiazole rings;
in the general formula III R 1 Selected from the group consisting of alkyl, aryl, alkanoyl, arylacyl, arylsulfonyl, alkylsulfonyl, alkyl groups containing a six-membered ring; wherein the alkyl in the alkyl, the alkyl acyl and the alkyl sulfonyl is the alkyl with 1-10 carbons;
in the general formula III, X is selected from oxygen or sulfur;
in the general formula III R 2 Selected from fluorine, chlorine, bromine or iodine;
in the general formula III R 3 Selected from alkoxy, alkylamide, ester, sulfonamide, acyl or sulfonyl; wherein the alkyl in the alkyl, alkylamide, alkylacyl and alkylsulfonyl is alkyl with 1-10 carbons;
in the general formula IV, A is selected from aromatic rings or aromatic heterocycles, including pyridine rings, benzene rings, thiophene rings or thiazole rings;
in the general formula IV, B is selected from pyridine rings or benzene rings;
r in the general formula IV 1 Selected from fluorine, chlorine, bromine, iodine, alkyl, alkoxy or acyl;
x in the general formula IV is selected from oxygen or sulfur;
in the general formula IV, y is selected from 1 or 2;
r in the general formula IV 2 Selected from fluorine, chlorine, bromine or iodine;
r in the general formula IV 3 Selected from alkoxy, alkylamido, alkylester, alkylsulfonamido, alkanoyl or alkylsulfonyl; wherein the alkoxy group, the alkylamide group, the alkyl ester group, the alkylsulfonamide group, the alkylacyl group orThe alkyl group in the alkylsulfonyl group is an alkyl group of 1 to 10 carbons.
2. The compound of claim 1 as an antagonist of S1PR2, wherein the compound of formula I, II, III, IV has the structure:
Figure FDA0003852045030000021
Figure FDA0003852045030000031
Figure FDA0003852045030000041
3. a pharmaceutical composition comprising a compound of any one of claims 1-2, a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, and a pharmaceutically acceptable excipient.
4. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, for use as an S1PR2 antagonist.
5. Use of a pharmaceutical composition according to claim 3 as an S1PR2 antagonist.
6. Use of a compound of claim 1 or 2, or a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, for the manufacture of a medicament for the inhibition of expression of MMP2, IL-5, TGF- β 1, and increased expression of BMPR 2.
7. Use of a compound of claim 1 or 2, or a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer, or geometric isomer thereof, in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder, or condition that benefits from inhibition of expression of MMP2, IL-5, TGF- β 1, increased expression of BMPR 2.
8. Use of a compound according to claim 1 or 2, or a pharmaceutically acceptable salt, solvate, prodrug, active metabolite, N-oxide, stereoisomer, tautomer or geometric isomer thereof, for the manufacture of a medicament for the prophylaxis and/or treatment of pulmonary hypertension, pulmonary stenosis, chronic obstructive pulmonary disease, chronic tracheitis.
9. The pharmaceutical composition of claim 3, for use in the preparation of a medicament for the prevention and/or treatment of pulmonary hypertension, pulmonary stenosis, chronic obstructive pulmonary disease, chronic tracheitis.
CN202211135871.5A 2022-09-19 2022-09-19 S1PR2 antagonist and application thereof in preparation of medicines for treating lung diseases Active CN115433107B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202211135871.5A CN115433107B (en) 2022-09-19 2022-09-19 S1PR2 antagonist and application thereof in preparation of medicines for treating lung diseases

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202211135871.5A CN115433107B (en) 2022-09-19 2022-09-19 S1PR2 antagonist and application thereof in preparation of medicines for treating lung diseases

Publications (2)

Publication Number Publication Date
CN115433107A true CN115433107A (en) 2022-12-06
CN115433107B CN115433107B (en) 2024-06-18

Family

ID=84248743

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202211135871.5A Active CN115433107B (en) 2022-09-19 2022-09-19 S1PR2 antagonist and application thereof in preparation of medicines for treating lung diseases

Country Status (1)

Country Link
CN (1) CN115433107B (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103958465A (en) * 2011-09-29 2014-07-30 小野药品工业株式会社 Phenyl derivative
CN107849038A (en) * 2015-06-01 2018-03-27 达尔豪西大学 S1PR2 antagonists and application thereof
US20200369626A1 (en) * 2019-04-30 2020-11-26 Washington University Compositions of sphingosine-1-phosphate receptor 2 (s1pr2) binding agents and uses thereof
WO2021026479A1 (en) * 2019-08-07 2021-02-11 Stamford, Andrew Small molecule inhibitors of s1p2 receptor and uses thereof
CN112759564A (en) * 2019-11-05 2021-05-07 中国医学科学院药物研究所 Diaryl urea compound and its preparation method and medicinal use
CN113135909A (en) * 2020-01-17 2021-07-20 北京深蓝泰医药科技有限公司 DPD inhibitor and preparation method, pharmaceutical composition and application thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103958465A (en) * 2011-09-29 2014-07-30 小野药品工业株式会社 Phenyl derivative
CN107849038A (en) * 2015-06-01 2018-03-27 达尔豪西大学 S1PR2 antagonists and application thereof
US20200369626A1 (en) * 2019-04-30 2020-11-26 Washington University Compositions of sphingosine-1-phosphate receptor 2 (s1pr2) binding agents and uses thereof
WO2021026479A1 (en) * 2019-08-07 2021-02-11 Stamford, Andrew Small molecule inhibitors of s1p2 receptor and uses thereof
CN112759564A (en) * 2019-11-05 2021-05-07 中国医学科学院药物研究所 Diaryl urea compound and its preparation method and medicinal use
CN113135909A (en) * 2020-01-17 2021-07-20 北京深蓝泰医药科技有限公司 DPD inhibitor and preparation method, pharmaceutical composition and application thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DONGDONG LUO ET AL.: "Rational Design, Synthesis, and Biological Evaluation of Novel S1PR2 Antagonists for Reversing 5‑FU-Resistance in Colorectal Cancer", 《J. MED. CHEM.》, vol. 65, pages 14553 *
KENSUKE KUSUMI ET AL.: "Discovery of novel S1P2 antagonists, part 3: Improving the oral bioavailability of a series of 1, 3-bis(aryloxy)benzene derivatives", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》, vol. 26, pages 1209, XP029409769, DOI: 10.1016/j.bmcl.2016.01.031 *
ZONGHUA LUO ET AL.: "Design, synthesis, and in vitro bioactivity evaluation of fluorine-containing analogues for sphingosine-1-phosphate 2 receptor", 《BIOORGANIC & MEDICINAL CHEMISTRY》, vol. 27, pages 3619, XP085746100, DOI: 10.1016/j.bmc.2019.06.047 *

Also Published As

Publication number Publication date
CN115433107B (en) 2024-06-18

Similar Documents

Publication Publication Date Title
EP3280707B1 (en) Bicyclic quinazolinone derivatives
EP2068878B1 (en) Rho kinase inhibitors
ES2298288T3 (en) DERIVATIVES OF 4,6-DIFENYL PIRIDINE AS ANTI-INFLAMMATORY AGENTS.
US20220306589A1 (en) Triazine derivatives for treating diseases relating to neurotrophins
WO2014181287A1 (en) Heterocyclyl compounds and uses thereof
KR20140067048A (en) Lysophosphatidic acid receptor antagonists
JP2003012653A (en) Quinazoline derivative
KR20190003685A (en) Inhibitors of indoleamine 2,3-dioxygenase and methods of use thereof
EP4079748A1 (en) Modulators of sortilin activity
CN117642416A (en) Sortilin activity modulators
US20190367500A1 (en) Quinazoline derivative
JP6197971B1 (en) KCNQ2-5 channel-related disease prevention and / or treatment agent
CN114127054A (en) N-methyl, N- (6- (methoxy) pyridazin-3-yl) amine derivatives as Autotaxin (ATX) modulators for the treatment of inflammatory airways or fibrotic diseases
CN112513032A (en) 4-substituted phenyl-1, 3, 5-triazine derivatives as modulators of TRK receptors
US20090099243A1 (en) Organic compounds
WO2021127459A1 (en) Gpr139 receptor modulators
CN115433107B (en) S1PR2 antagonist and application thereof in preparation of medicines for treating lung diseases
KR20110045095A (en) Pyridine Derivatives, a VFR-2 Receptor and Protein Tyrosine Kinase Inhibitor
WO2021149767A1 (en) Heterocyclic derivative
WO2005028453A1 (en) Phenylacetic acid derivative, process for producing the same, and use
EP4032891A1 (en) Pyridine derivatives as modulators of sortilin activity
JPWO2017183723A1 (en) KCNQ2-5 channel activator
US20240076272A1 (en) Activators of Heme Regulated Inhibitor Kinase (HRI)
US20240076284A1 (en) New compounds and methods
WO2022129914A1 (en) Alkyne derivatives as inhibitors of c-abl

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant