WO2005103030A1 - Composes de benzene a di-substitution en positions 1,4, leur procede de preparation et leur utilisation - Google Patents

Composes de benzene a di-substitution en positions 1,4, leur procede de preparation et leur utilisation Download PDF

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WO2005103030A1
WO2005103030A1 PCT/CN2005/000029 CN2005000029W WO2005103030A1 WO 2005103030 A1 WO2005103030 A1 WO 2005103030A1 CN 2005000029 W CN2005000029 W CN 2005000029W WO 2005103030 A1 WO2005103030 A1 WO 2005103030A1
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compound
solvent
cypa
substituted
concentration
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Hualiang Jiang
Xu Shen
Jianhua Shen
Hong Liu
Xiaomin Luo
Chunshan Gui
Jian Li
Hongxia Guo
Chenjing Li
Kaixian Chen
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Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences
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    • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
    • C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
    • C07C69/84—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring of monocyclic hydroxy carboxylic acids, the hydroxy groups and the carboxyl groups of which are bound to carbon atoms of a six-membered aromatic ring
    • C07C69/86—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring of monocyclic hydroxy carboxylic acids, the hydroxy groups and the carboxyl groups of which are bound to carbon atoms of a six-membered aromatic ring with esterified hydroxyl groups
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    • 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
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    • 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
    • C07D213/24—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 substituted hydrocarbon radicals attached to ring carbon atoms
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/04—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to the ring carbon atoms
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    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/12—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms 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
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    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
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    • 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
    • C07D401/10—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 carbon chain containing aromatic rings
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    • 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
    • C07D401/14—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 three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/10—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing aromatic rings

Definitions

  • the present invention relates to a class of 1,4-disubstituted benzene compounds, and in particular to a class of 1,4-disubstituted benzene compounds that are small molecule non-peptide ligands of cyclophilin A, which compounds can be used as immunosuppressive agents.
  • the invention also relates to a method for preparing such compounds. Background technique
  • CsA Cyclosporine A
  • the first generation represented by corticosteroids, azathioprine, and anti-lymphocyte globulin (ALG), whose main role is to lyse immune-active cells and block cell differentiation, which is characterized by non-specific and extensive immune suppression;
  • Generation represented by cyclosporine and FK506, which block the effector interleukin 2 (IL-2) of immunocompetent cells, and are relatively specific because they are mainly lymphocytes.
  • Clone antibodies, cidolimus, and mycophenolate lipids are representative, which act on antigen presentation and intermolecular interactions, and have synergistic effects with second-generation preparations; fourth generation: anti-IL-2 receptor monoclonal antibodies FTY20, etc. are mainly aimed at changing the cytokine environment, such as inhibiting TH1 and enhancing TH2.
  • Another object of the present invention is to provide the compound of the present invention as a small molecule non-peptide ligand of cyclophilin A, used as an immunization inhibitor and a drug for preventing transplant rejection, host disease and autoimmune disease caused by transplantation.
  • the compound of the present invention is a 1,4-disubstituted benzene compound or a pharmaceutically acceptable salt thereof having the structure of the following general formula I:
  • X and Z are each independently selected from 0, S or NH;
  • Y is selected from CO or S0 2 ;
  • R 2 is selected from the following structural groups (11, III, IV or V):
  • n 0, 1, 2 or 3;
  • R 3 and R 4 are each independently selected from hydrogen; -C 6 linear or branched alkyl, alkenyl, or alkynyl; nitro; halogen: cyano; trifluoromethyl; trifluoromethoxy;
  • R 5 each independently selected from hydrogen; methyl; ethyl; cyclohexyl; aromatic Ar; 5-7 membered heteroaryl or substituted heteroaryl containing 1-3 heteroatoms selected from oxygen, sulfur or nitrogen Group, wherein the heteroaryl group may form a condensed ring aryl group with a phenyl group or a 5-7 membered heterocyclic aryl group, and the substituent in the substituted heteroaryl group is arbitrarily selected from one or 2-5 of the following groups: halogen; -3 ⁇ 4 linear or branched alkyl, alkenyl or alkynyl; cyano; nitro; amino; hydroxyl; methylol; trifluoromethyl; trifluoromethoxy; carboxyl; -C 4 alkoxy Thiol d-acyl aryl Ar
  • the aromatic group Ar refers to a phenyl group, a naphthyl group, a biphenyl group, or a substituted phenyl group, wherein the substituents in the substituted phenyl group are arbitrarily selected from 1-4 of the following groups: halogen; dC 6 straight or branched chain Alkyl, alkenyl or alkynyl; cyano; nitro; amino; hydroxyl; methylol; trifluoromethyl; trifluoromethoxy; carboxyl;-alkoxy; mercapto;-acyl.
  • compositions such as propionic acid, oxalic acid, malonic acid, ruberic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, and citric acid.
  • organic acids such as propionic acid, oxalic acid, malonic acid, ruberic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, and citric acid.
  • an acidic amino 'acid such as aspartic acid and glutamic acid
  • an inorganic base such as sodium, potassium, calcium, aluminum, and ammonium salts
  • an organic base such as formazan Amine salt, ethyl
  • a preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound or a pharmaceutically acceptable salt thereof having the structure of the general formula I:
  • Another preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • Yet another preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having a structure of general formula I or a pharmaceutically acceptable salt thereof-wherein X and Z are 0; Y is CO;
  • R 2 is the same as that of the aforementioned compound.
  • Another preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I, or a compound thereof Pharmaceutically acceptable salts:
  • Ri and R 2 are the same as those of the aforementioned compounds.
  • the fifth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • a sixth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof-wherein X is NH; Z is 0; Y is CO;
  • Ri, R 2 is the same as that of the aforementioned compound.
  • the seventh preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having a structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • An eighth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having a structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • Ri and R 2 are the same as those of the aforementioned compounds.
  • the ninth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • R [> R 2 has the same definition as the aforementioned compound.
  • the tenth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • the eleventh preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • the twelfth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • Ri is the same as that of the aforementioned compound.
  • the thirteenth preferred compound of the compound of the present invention is a 1,4-disubstituted harmless compound having the structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • Ri is the same as that of the aforementioned compound
  • R 2 is a group having the following structural formula:
  • R 3 , R 5 are the same as the definitions of the aforementioned compounds.
  • the fourteenth preferred compound of the compound of the present invention is 1, 4- having a structure of the general formula I: a pharmaceutically acceptable salt thereof:
  • R has the same definition as the aforementioned compound
  • R 6 is the same as that of the aforementioned compound.
  • the fifteenth preferred compound of the compound of the present invention is a 1,4-disubstituted benzene compound having a structure of the general formula I or a pharmaceutically acceptable salt thereof:
  • R has the same definition as the aforementioned compound
  • R 2 is a group having the following structural formula ⁇
  • n 0, 1, 2 or 3;
  • Ar has the same definition as the aforementioned compound.
  • the compound of the present invention is Z-acylated and deprotected from the compound R'- X-Ph-Y-Cl (see editor Ji Zhizhong: Chemical Pharmaceutical Technology, Beijing: China Medical Science and Technology Press, 1997, p. 173-175), X-acylation (see Giuseppe AM, Henry MS, J. Med. Chem., 1997, 40, 1794-1807) prepared by the operation of the tritium reaction.
  • R ' is acetyl or benzyl: X, Y, Z, R !, R 2 are as defined above.
  • the inert solvents used in the acylation reaction include methylene chloride, 1,2-dichloroethane, benzene, toluene, chloroform, carbon tetrachloride, tetrahydrofuran, or 1,4-dioxane.
  • the chlorine-containing substitutes used in the preparation of the acid chloride (Cl-R 2 ) include dichlorosulfoxide, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, oxalyl chloride, triphenylphosphine chloride, or cyanuric chloride.
  • the reaction temperature and reaction time depend on the reaction of the specific compound.
  • the post-treatment methods generally used after the completion of each step of the reaction include extraction, filtration, evaporation of the solvent under reduced pressure, and purification to obtain a pure product, among which purification methods include column chromatography or recrystallization.
  • the final product I was confirmed by melting point, nuclear magnetic resonance or mass spectrometry data.
  • Class I A compounds are prepared by using p-acetylaminobenzenesulfonyl chloride as the raw material, which is respectively subjected to sulfonylation, deprotection and N-acylation.
  • the specific synthesis process is carried out in the following three steps:
  • Substituted amines are reacted with p-acetylaminobenzenesulfonyl chloride in alkaline solutions at room temperature.
  • the solvents used include pyridine, triethylamine, 4-dimethylaminopyridine (DMAP), diisopropylethylamine or inorganic bases.
  • An aqueous solution in which the inorganic base used includes sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium hydroxide, potassium hydroxide. The temperature is between 40-45 ° C. Stir for 12-36 hours. The reaction solution was poured into water, and the aqueous layer was extracted with an organic solvent.
  • the organic solvents used included ethyl acetate, dichloromethane, dichloroethane, ether, and chloroform.
  • the separated organic layer was dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure.
  • the residue was subjected to column chromatography to obtain 4-acetylaminobenzenesulfonamide. Put this sulfonamide into a 10% sodium hydroxide aqueous solution and reflux for 2 hours, control the temperature of the reaction solution between 70-80 ° C, adjust the pH to 1 with concentrated hydrochloric acid, and precipitate a large amount of solids. After suction filtration, water washing and drying, Product VI.
  • the compound VII dissolved in an inert solvent is reacted with an excess of a chlorine-containing substitute, wherein the chlorine-containing substitute includes dichlorosulfoxide, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, chlorochlor, triphenylphosphine Chloride or cyanuric chloride; inert solvents include dichloromethane, 1,2-dichloroethane, benzene, toluene, chloroform, carbon tetrachloride, tetrahydrofuran, 1,4-dioxane.
  • the chlorine-containing substitute includes dichlorosulfoxide, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, chlorochlor, triphenylphosphine Chloride or cyanuric chloride
  • inert solvents include dichloromethane, 1,2-dichloroethane, benzene, tolu
  • class IB compounds are based on ⁇ -bromomethyl ketone, which is obtained through condensation, ring expansion, halogenation and N-acylation.
  • the specific synthesis process is carried out in the following three steps:
  • the obtained solid was dispersed into powder with diethyl ether and suction filtered.
  • the solvent was distilled off under reduced pressure, and the solid was washed with ethyl acetate and then purified by recrystallization to obtain compound VIII.
  • I react a compound soluble in an inert solvent with an excess of a chlorine-containing replacement, which includes dichlorosulfoxide, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, oxalyl chloride, triphenylphosphine chloride Compounds or cyanuric chloride; inert solvents include dichloromethane, 1,2-dichloroethane, benzene, toluene, chloroform, carbon tetrachloride, tetrahydrofuran, 1,4-dioxane.
  • a chlorine-containing replacement which includes dichlorosulfoxide, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, oxalyl chloride, triphenylphosphine chloride
  • inert solvents include dichloromethane, 1,2-dichloroethane, benzene, toluene
  • Ic compounds are synthesized by using various aralkylcarboxylic acids as raw materials, which are respectively halogenated and N-acylated.
  • the specific synthesis process is carried out in the following two steps ⁇
  • aralkyl carboxylic acid dissolved in an inert solvent with an excess of a chlorine-containing substitute
  • the chlorine-containing substitute includes dichlorosulfoxide, phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, oxalyl chloride, Triphenylphosphine chloride or cyanuric chloride
  • inert solvents include dichloromethane, 1,2-dichloroacetamidine, benzene, toluene, chloroform, carbon tetrachloride, tetrahydrofuran, 1,4-dioxane.
  • the compounds of the present invention can be used as small-molecule non-peptide ligands of cyclophilin A, and can be used to induce immunosuppressive effects and treat transplant rejection by blocking the effect link of interleukin 2 (IL-2) of immunocompetent cells Response, treatment of host diseases and autoimmune diseases caused by transplantation.
  • IL-2 interleukin 2
  • the action mechanism of the compounds of the present invention to induce immunosuppression is as follows: First, it is combined with cyclophilin A (CypA) to form a complex CypA-CsA, which is combined with its effector protein calcineurin to inhibit serine / threonine Acid phosphatase activity, thereby blocking the production of some cytokines, including interleukin-2.
  • CypA cyclophilin A
  • the compounds of the present invention have the advantages of small molecules, non-peptides, easy synthesis, low toxicity, and the like.
  • the method for preparing the compound of the present invention has the advantages of mild reaction conditions, abundant raw materials, easy operation and simple post-treatment. 3.
  • the compound of the present invention has a large binding constant K D with cyclophilin A, which proves that the two have strong binding ability. Therefore, the compound of the present invention has a good mechanism of inducing immunosuppression, and can be used as an immunization inhibitor. And drugs for transplant rejection, host disease and autoimmune diseases caused by transplantation. BRIEF DESCRIPTION OF THE DRAWINGS
  • Figure 1 shows the kinetic curve of the binding of compound I A -1 to CypA, where ai is the kinetic curve when the concentration of I A -1 is 1 X 10 " 5 M, and ⁇ is the concentration of I A -1 is 5 X 1 ( kinetic curves of ⁇ 6 ⁇ , Cl is the concentration of I a -1 kinetic curve, d! a -1 is the concentration I kinetics 1.25 X 10- 6 M when the time of 2.5 X 10 ⁇ ⁇ ⁇ , ei is the kinetic curve when the concentration of I A -1 is 6.25 X 10 " 7 M, is the kinetic curve when the concentration of I A -1 is 3.125 X 1 (T 7 M, gl is the working baseline.
  • Figure 2 shows the kinetic curve of the binding of compound Ic-3 to CypA, where a 2 is the kinetic curve when the concentration of I c -3 is 1 X 10 " 5 M, and b 2 is the concentration of I c -3 is 5 X 10_ kinetic curves of ⁇ ⁇ , c 2 is the I c - 3 concentration kinetics 2.5 X 10- S M during, d 2 is the concentration of I c -3 kinetics 1.25 X 10- 5 M when, e 2 is the concentration of I c -3 kinetics 6.25 X 10 "7 M when, f 2 is the I c -3 at a concentration of 3.125 X 10- 7 M kinetics when, g 2 work baseline.
  • Figure 3 shows the effect of DMSO on CypA fluorescence, where CypA concentration is maintained at 6 ⁇ M, a 3 is the fluorescence intensity curve of CypA at a DMSO concentration of 0 ⁇ M, b 3 is the fluorescence intensity curve of CypA at a DMSO concentration of 2 ⁇ M, and c 3 is a DMSO concentration of 4 ⁇ M CypA fluorescence intensity curve at time, d 3 is CypA fluorescence intensity curve at DMSO concentration of 8 ⁇ M, e 3 is CypA fluorescence intensity curve at DMSO concentration of 16 ⁇ M, f 3 is CypA fluorescence intensity curve at DMSO concentration of 32 ⁇ M, and g 3 is DMSO CypA fluorescence intensity curve at a concentration of 64 ⁇ M. ⁇
  • Figure 4 shows the effect of irrelevant compounds on CypA fluorescence, where CypA concentration is maintained at 6 ⁇ M, which is the fluorescence intensity curve of CypA at the compound concentration of 0 ⁇ M, b 4 is the fluorescent intensity curve of CypA at the compound concentration of 2 ⁇ M, and c 4 is the compound The fluorescence intensity curve of CypA at a concentration of 4 ⁇ M, d 4 is the fluorescence intensity curve of CypA at a concentration of 8 ⁇ M, e 4 is the fluorescence intensity curve of CypA at a concentration of 16 ⁇ M, and f 4 is the fluorescence intensity curve of CypA at a concentration of 32 ⁇ M. And g 4 are the fluorescence intensity curves of CypA at a concentration of 64 ⁇ M of the compound.
  • Figure 5 shows the effect of compound I A -1 on the fluorescence quenching of CypA, where CypA concentration is maintained at 6 ⁇ M, a 5 is the fluorescence intensity curve of CypA at a concentration of -1 at 0 ⁇ M, and b 5 is the CypA at a concentration of 2 ⁇ M at I A -1 Fluorescence intensity curve, C5 is the fluorescence intensity curve of CypA at a concentration of 4 ⁇ M at -1, d 5 is the fluorescence intensity curve of CypA at a concentration of I A -1 at 8 ⁇ M, e 5 is the fluorescence intensity curve of CypA at a concentration of 16 ⁇ M at I A -1, f 5 is a fluorescence intensity curve of CypA at a concentration of I A -1 at 32 ⁇ M, and g 5 is a fluorescence intensity curve of CypA at a concentration of -1 at 64 ⁇ M.
  • FIG. 6 shows a linear fitting graph of AFc vs. AFc / C (xlO 6 ) obtained from the fluorescence quenching experiment of compound I A -1 on CypA.
  • Figure 7 shows the effect of compound I A -2 on the fluorescence quenching of CypA, where CypA concentration is maintained at 6 ⁇ M, a 5 is the fluorescence intensity curve of CypA at IA-2 concentration of 0 ⁇ M, and b 6 is CypA at I A -2 concentration of 2 ⁇ M.
  • the fluorescence intensity curves c 6 I a -2 is the concentration at which the fluorescence intensity curves 4 ⁇ CypA
  • d 6 is the fluorescence intensity when I a -2 CypA 8 ⁇ concentration curve
  • e 6 concentration is I a -2 16 ⁇ fluorescence when CypA
  • the intensity curve, f 6 is the fluorescence intensity curve of CypA at a concentration of I A -2 of 32 ⁇ M
  • g 6 is the fluorescence intensity curve of CypA at a concentration of I A -2 of 64 ⁇ M.
  • Figure 9 shows the effect of compound I c -3 on the fluorescence quenching of CypA, where the CypA concentration is maintained at 6 ⁇ M, a 7 is the fluorescence intensity curve of CypA at the concentration of I A -3 0 ⁇ M, and b 7 is the concentration of I A -3 at 2 ⁇ M CypA fluorescence intensity curve, c 7 is the fluorescence intensity curve of CypA at a concentration of 4 ⁇ M in I A -3, d 7 is the fluorescence intensity curve of CypA at a concentration of 3 ⁇ M at 8 ⁇ M, and e 7 is the fluorescence intensity curve of CypA at a concentration of 16 ⁇ M at IA-3. , F 7 is the fluorescence intensity curve of CypA at a concentration of I A -3 of 32 ⁇ M, and g 7 is the fluorescence intensity curve of CypA at a concentration of ⁇ 3 of 64 ⁇ M.
  • Figure 10 shows compounds I A -3 quenching of the fluorescence of CypA on AFc AFc / C (xlO 6) obtained by linear fitting experimental FIG.
  • Figure 11 shows the effect of compound I A -4 on the fluorescence quenching of CypA, where the CypA concentration is maintained at 6 ⁇ M, a 8 is the -4 concentration of 0 ⁇ M, CypA fluorescence intensity curve, and b 8 is the CypA at I A -4 concentration of 2 ⁇ M.
  • c 8 is I a - 4 fluorescence intensity curves of concentration 4 ⁇ CypA
  • d 8 when the fluorescence intensity I a -4 CypA 8 ⁇ concentration curve
  • e s is the fluorescence intensity when I a -4 CypA concentration 16 ⁇ curve
  • I a -4 32 ⁇ concentration fluorescence intensity curves of CypA when I a -4 CypA 64 ⁇ concentration curve.
  • FIG 12 shows compound I A -4 CypA fluorescence quenching of AFc AFc / C (xl O 6) linear fitting of FIG experiments.
  • FIG. 13 shows the effect of compound I c -1 on the fluorescence quenching of CypA, where CypA concentration is maintained at 6 ⁇ M, a 9 is the fluorescence intensity curve of CypA at I c -1 concentration of 0 ⁇ M, and b 9 is the I c -1 concentration at 2 ⁇ M.
  • CypA fluorescence intensity curve ⁇ is the CypA fluorescence intensity curve at an Ic-1 concentration of 4 ⁇ M
  • d 9 is the CypA fluorescence intensity curve at an I c -1 concentration of 8 ⁇ M
  • e 9 is the CypA fluorescence intensity curve at an Ic-1 concentration of 16 ⁇ M
  • 3 ⁇ 4 is the fluorescence intensity curve of CypA at an I c -1 concentration of 32 ⁇ M
  • g 9 is the fluorescence intensity curve of CypA at an I c -1 concentration of 64 ⁇ M.
  • FIG. 14 shows a linear fitting graph of AFc versus AFc / C (xlO 6 ) obtained from the fluorescence quenching experiment of Compound Ic-1 on CypA.
  • Figure 15 shows the CypA compound I c 3 on the fluorescence quenching of CypA influence diagram, wherein the concentration of CypA maintained 6 ⁇ , a 10 as fluorescence intensity curves I c -3 concentration of CypA ⁇ , I c is the concentration 2 ⁇ -3 Fluorescence intensity curve Line, c 1 () is the I c - fluorescence intensity curves 4 ⁇ CypA concentration of 3, d 1 () is the fluorescence intensity I c -3 8 ⁇ CypA concentration curve, e l 0 is the concentration of I c -3 16 ⁇ when CypA The fluorescence intensity curve, f w is the fluorescence intensity curve of CypA at an I c -3 concentration of 32 ⁇ M, and g 1 () is the fluorescence intensity curve of CypA at an I c -3 concentration of 64 ⁇ M.
  • FIG. 16 shows a linear fit graph of AFc vs. AFc / C (xlO 6 ) obtained from the fluorescence quenching experiment of compound I c -3 on CypA. detailed description
  • the melting point was measured with a SGW-X-4 micro melting point apparatus (Nanjing Kejie Instrument Application Research Institute), and the temperature was uncorrected; nuclear magnetic resonance was measured by a Bmker AMX-400 nuclear magnetic resonance apparatus (Bmker) TMS (tetramethylsilane) was the internal standard; mass spectra were determined by MAT-711 and MAT-95 mass spectrometers (Finnigan).
  • the product is mainly purified by column chromatography.
  • the silica gel is 200-300 mesh (produced by Qingdao Ocean Chemical Plant).
  • the obtained solution was slowly added dropwise to 4-amino-N- (4-methyl-2-pyrimidine under stirring at room temperature. Group) -benzenesulfonamide in a pyridine solution (0.5g / 5ml). After the dropwise addition is completed, the temperature of the reaction solution is controlled between 40-45 ° C and stirred for 24 hours.
  • the reaction solution was poured into 50 ml of water, and extracted with dichloroethane (20mk3). The organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography, and the eluent was ethyl acetate.
  • 4-amino-N- (2-quinoxalinyl) was prepared from 2.34 g of p-acetylaminobenzenesulfonyl chloride and 1.45 g of 2-amino-quinoxaline.
  • -Benzenesulfonamide 1.08 g as a pale yellow solid with a yield of 31.6%.
  • the reaction solution was poured into 50 ml of water, and extracted with dichloroethane (30 ml x 3). The organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography, and the eluent was acetic acid. Ethyl ester / petroleum ether (1: 2, v / v) to obtain the target 4- (4-chlorophenylacetyl) amino-N- (2-quinoxalinyl) benzenesulfonamide 0.15g The yield was 33.1%.
  • 2- (2-methoxyphenyl) quinoline was obtained from the reaction product of 1 equivalent of 2,3-dione indole and 1.2 equivalent of 2-methoxyacetophenone.
  • MS-EI 514 (M) 234 (100%).
  • Example 7 Preparation of 4- [2- (3-bromophenyl) quinoline-4-acyl] amino-N- (2-sleepoxolinyl) benzenesulfonamide (I A -5)
  • Example 18 Preparation of 4- (2,4-dichlorobenzoyl) amino-N- (2-quinoxalinyl) benzenesulfonamide (I c -3) According to the same method as described in Example 3, Reaction product of 1 equivalent of 2,4-dichlorobenzoic acid with an excess of dichlorosulfoxide 2,4-dichlorobenzoyl chloride and 1 equivalent of 2-amino-quinoxaline with 1 equivalent of p-acetylaminobenzene The reaction product of sulfonyl chloride, 4-amino-N- (2-quinoxalinyl) benzenesulfonamide was linked to prepare I c -3 as a pale yellow solid.
  • HBS-EP buffer (10 mM Hepes, 150 mM NaCl, 3.4 mM EDTA, 0.005% (v / v) surfactant P20, pH 7.4)
  • CM5 chip purchased from BIACORE AB (Uppsala, Sweden)
  • BIAcore3000 Biosensor using surface plasmon resonance technology (BIACORE AB, Uppsala, Sweden)
  • CypA is coupled to the CM5 chip by amino coupling method (Amino coupling method: EDC and NHS are mixed in equal volumes to activate the carboxymethyl groups on the surface of the CM5 chip to become an intermediate state that is easy to react with amino groups, and then Purified CypA was injected onto the chip's surface. The amino group at the N-terminus of the protein was covalently bonded to the intermediate state of the carboxyl group through a shrinkage reaction to couple CypA to the chip surface. active. Test compound dissolved in dimethylsulfoxide (DMSO), and diluted with HBS-EP buffer to lxlO '5 -.
  • DMSO dimethylsulfoxide
  • the change in CypA fluorescence is related to the concentration of the compound bound to CypA, so the change in fluorescence can indirectly reflect the concentration of the compound bound to CypA, and the equilibrium dissociation constant of the interaction between the compound and CypA can be calculated by the formula.
  • test compound I A -1, I A -2, I A -3, I A -4, Ic-K I c -3, the test compound is dissolved with DMSO, the concentration is 10mM;
  • Negative reference substance dimethyl sulfoxide (DMSO); other unrelated compounds, that is, compounds designed for other disease targets with similar molecular weight as the compound of the present invention
  • Tris-HCl buffer 20mM Tris-HCl, pH7.4; 100mM NaCl
  • the fluorescence change value after deducting the influence of DMSO is sorted, and AFc / C (the ratio of the fluorescence change value of the protein after the compound is added to the compound concentration) is used as the abscissa, and AFc (the fluorescence change value of the protein after the compound is added) is used as the ordinate.
  • the obtained slope is the equilibrium dissociation constant K d of CypA and the small molecule compound.
  • CypA concentration was maintained at 6 ⁇ M, and the fluorescence value of CypA was measured at a compound concentration of 0 ⁇ M, 2 ⁇ M, 4 ⁇ M, 8 ⁇ M, 16 ⁇ M, 32 ⁇ M, and 64 ⁇ M, respectively.
  • the effect of this compound on CypA fluorescence is shown in Figure 4.
  • the value is equal to AFc, which indicates the fluorescence change value of the protein after adding the compound, that is, the difference between the Fc value after adding the compound and the Fc without adding the compound.
  • AFc indicates the fluorescence change value of the protein after adding the compound, that is, the difference between the Fc value after adding the compound and the Fc without adding the compound.

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Abstract

La présente invention concerne des composés de benzène à di-substitution en positions 1,4 à petites molécules utilisés comme ligands non peptidiques de cyclophiline A, lesquels ont la structure selon la formule générale (I). Des tests d'activité biologique ont permis d'indiquer que les composés de la présente invention présentent une capacité de combinaison relativement forte avec la cyclophiline A, il est ainsi prouvé que le mécanisme d'action permet d'induire une immunodépression. La présente invention concerne également le procédé de préparation desdits composés.
PCT/CN2005/000029 2004-02-27 2005-01-10 Composes de benzene a di-substitution en positions 1,4, leur procede de preparation et leur utilisation WO2005103030A1 (fr)

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US10189788B2 (en) * 2014-09-09 2019-01-29 Bayer Pharma Aktiengesellschaft Substituted N,2-diarylquinoline-4-carboxamides and the use thereof as anti-inflammatory agents
US10266488B2 (en) 2013-10-10 2019-04-23 Eastern Virginia Medical School 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide derivatives as potent and selective inhibitors of 12-lipoxygenase

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CN101555212B (zh) * 2009-05-22 2012-06-20 戴立忠 能输送生物活性物质的合成小分子化合物及其应用

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WO2001023378A1 (fr) * 1999-09-28 2001-04-05 Applied Research Systems Ars Holding N.V. Derives sulfonamides actifs sur le plan pharmacologique
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10266488B2 (en) 2013-10-10 2019-04-23 Eastern Virginia Medical School 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide derivatives as potent and selective inhibitors of 12-lipoxygenase
US10752581B2 (en) 2013-10-10 2020-08-25 Eastern Virginia Medical School 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide derivatives as potent and selective inhibitors of 12-lipoxygenase
US11274077B2 (en) 2013-10-10 2022-03-15 Eastern Virginia Medical School 4-((2-hydroxy-3-methoxybenzyl)amino)benzenesulfonamide derivatives as potent and selective inhibitors of 12-lipoxygenase
US10189788B2 (en) * 2014-09-09 2019-01-29 Bayer Pharma Aktiengesellschaft Substituted N,2-diarylquinoline-4-carboxamides and the use thereof as anti-inflammatory agents
US10479765B2 (en) 2014-09-09 2019-11-19 Bayer Pharma Aktiengesellschaft Substituted N,2-diarylquinoline-4-carboxamides and the use thereof as anti-inflammatory agents

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