CN108358853A - 一种抗神经炎症的二苯乙烯类似物及其制备方法和应用 - Google Patents
一种抗神经炎症的二苯乙烯类似物及其制备方法和应用 Download PDFInfo
- Publication number
- CN108358853A CN108358853A CN201810134754.4A CN201810134754A CN108358853A CN 108358853 A CN108358853 A CN 108358853A CN 201810134754 A CN201810134754 A CN 201810134754A CN 108358853 A CN108358853 A CN 108358853A
- Authority
- CN
- China
- Prior art keywords
- neuroinflamation
- straight chain
- compound
- chain saturated
- halogen
- 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
Links
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical group C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 23
- 125000005843 halogen group Chemical group 0.000 claims abstract description 18
- 229910052736 halogen Inorganic materials 0.000 claims abstract description 14
- 150000002367 halogens Chemical class 0.000 claims abstract description 14
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 7
- HDWLUGYOLUHEMN-UHFFFAOYSA-N Dinobuton Chemical compound CCC(C)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1OC(=O)OC(C)C HDWLUGYOLUHEMN-UHFFFAOYSA-N 0.000 claims abstract description 4
- 241000425573 Talanes Species 0.000 claims abstract description 4
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 5
- -1 chloro, hydroxyl Chemical group 0.000 claims description 4
- 229940079593 drug Drugs 0.000 claims description 4
- 229910052731 fluorine Inorganic materials 0.000 claims description 4
- 239000011737 fluorine Substances 0.000 claims description 4
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 claims description 3
- 238000009833 condensation Methods 0.000 claims description 3
- 230000005494 condensation Effects 0.000 claims description 3
- 238000003684 Perkin reaction Methods 0.000 claims description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims description 2
- 238000006467 substitution reaction Methods 0.000 claims 6
- 150000001335 aliphatic alkanes Chemical class 0.000 claims 1
- 230000007131 anti Alzheimer effect Effects 0.000 claims 1
- 125000001153 fluoro group Chemical group F* 0.000 claims 1
- 150000003839 salts Chemical class 0.000 claims 1
- 229920006395 saturated elastomer Polymers 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 abstract description 44
- 230000008499 blood brain barrier function Effects 0.000 abstract description 10
- 210000001218 blood-brain barrier Anatomy 0.000 abstract description 9
- 238000000034 method Methods 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 8
- 208000024827 Alzheimer disease Diseases 0.000 abstract description 6
- 206010039966 Senile dementia Diseases 0.000 abstract description 4
- 150000002611 lead compounds Chemical class 0.000 abstract description 3
- RVFVPPONXUZVTJ-UHFFFAOYSA-N piperazine;pyrimidine Chemical compound C1CNCCN1.C1=CN=CN=C1 RVFVPPONXUZVTJ-UHFFFAOYSA-N 0.000 abstract description 3
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 112
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 28
- 238000005160 1H NMR spectroscopy Methods 0.000 description 28
- 239000007787 solid Substances 0.000 description 28
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 20
- FRJJJAKBRKABFA-TYFAACHXSA-N (4r,6s)-6-[(e)-2-[6-chloro-4-(4-fluorophenyl)-2-propan-2-ylquinolin-3-yl]ethenyl]-4-hydroxyoxan-2-one Chemical compound C(\[C@H]1OC(=O)C[C@H](O)C1)=C/C=1C(C(C)C)=NC2=CC=C(Cl)C=C2C=1C1=CC=C(F)C=C1 FRJJJAKBRKABFA-TYFAACHXSA-N 0.000 description 13
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 102000043136 MAP kinase family Human genes 0.000 description 12
- 108091054455 MAP kinase family Proteins 0.000 description 12
- 230000003110 anti-inflammatory effect Effects 0.000 description 12
- 239000000243 solution Substances 0.000 description 11
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 10
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 9
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 8
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 8
- 102000003777 Interleukin-1 beta Human genes 0.000 description 8
- 108090000193 Interleukin-1 beta Proteins 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 210000000274 microglia Anatomy 0.000 description 6
- 239000006180 TBST buffer Substances 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000037361 pathway Effects 0.000 description 5
- 0 CC1=CCC*1CC* Chemical compound CC1=CCC*1CC* 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 238000001994 activation Methods 0.000 description 4
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 4
- 239000011616 biotin Substances 0.000 description 4
- 229960002685 biotin Drugs 0.000 description 4
- 235000020958 biotin Nutrition 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 230000002757 inflammatory effect Effects 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000012491 analyte Substances 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- SNRUBQQJIBEYMU-UHFFFAOYSA-N dodecane Chemical compound CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 210000002682 neurofibrillary tangle Anatomy 0.000 description 3
- 230000026731 phosphorylation Effects 0.000 description 3
- 238000006366 phosphorylation reaction Methods 0.000 description 3
- 238000003825 pressing Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 2
- 208000037259 Amyloid Plaque Diseases 0.000 description 2
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000018745 NF-KappaB Inhibitor alpha Human genes 0.000 description 2
- 108010052419 NF-KappaB Inhibitor alpha Proteins 0.000 description 2
- QNVSXXGDAPORNA-UHFFFAOYSA-N Resveratrol Natural products OC1=CC=CC(C=CC=2C=C(O)C(O)=CC=2)=C1 QNVSXXGDAPORNA-UHFFFAOYSA-N 0.000 description 2
- 241000282898 Sus scrofa Species 0.000 description 2
- 241000282894 Sus scrofa domesticus Species 0.000 description 2
- LUKBXSAWLPMMSZ-OWOJBTEDSA-N Trans-resveratrol Chemical compound C1=CC(O)=CC=C1\C=C\C1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-OWOJBTEDSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229930016911 cinnamic acid Natural products 0.000 description 2
- 235000013985 cinnamic acid Nutrition 0.000 description 2
- 230000003920 cognitive function Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 2
- JBLLRCOZJMVOAE-HSQYWUDLSA-N n-[(2s)-1-[[(2s)-1-(1,3-benzothiazol-2-yl)-1-oxo-3-[(3s)-2-oxopyrrolidin-3-yl]propan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]-4-methoxy-1h-indole-2-carboxamide Chemical compound C([C@H](NC(=O)[C@H](CC(C)C)NC(=O)C=1NC=2C=CC=C(C=2C=1)OC)C(=O)C=1SC2=CC=CC=C2N=1)[C@@H]1CCNC1=O JBLLRCOZJMVOAE-HSQYWUDLSA-N 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 102000002574 p38 Mitogen-Activated Protein Kinases Human genes 0.000 description 2
- 108010068338 p38 Mitogen-Activated Protein Kinases Proteins 0.000 description 2
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000000770 proinflammatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 235000021283 resveratrol Nutrition 0.000 description 2
- 229940016667 resveratrol Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- SHAHPWSYJFYMRX-GDLCADMTSA-N (2S)-2-(4-{[(1R,2S)-2-hydroxycyclopentyl]methyl}phenyl)propanoic acid Chemical compound C1=CC([C@@H](C(O)=O)C)=CC=C1C[C@@H]1[C@@H](O)CCC1 SHAHPWSYJFYMRX-GDLCADMTSA-N 0.000 description 1
- KAFZOLYKKCWUBI-HPMAGDRPSA-N (2s)-2-[[(2s)-2-[[(2s)-1-[(2s)-3-amino-2-[[(2s)-2-[[(2s)-2-(3-cyclohexylpropanoylamino)-4-methylpentanoyl]amino]-5-methylhexanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]butanediamide Chemical compound N([C@@H](CC(C)C)C(=O)N[C@@H](CCC(C)C)C(=O)N[C@@H](CN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(N)=O)C(N)=O)C(=O)CCC1CCCCC1 KAFZOLYKKCWUBI-HPMAGDRPSA-N 0.000 description 1
- LJIOTBMDLVHTBO-CUYJMHBOSA-N (2s)-2-amino-n-[(1r,2r)-1-cyano-2-[4-[4-(4-methylpiperazin-1-yl)sulfonylphenyl]phenyl]cyclopropyl]butanamide Chemical compound CC[C@H](N)C(=O)N[C@]1(C#N)C[C@@H]1C1=CC=C(C=2C=CC(=CC=2)S(=O)(=O)N2CCN(C)CC2)C=C1 LJIOTBMDLVHTBO-CUYJMHBOSA-N 0.000 description 1
- FOLCUFKJHSQMEL-BIXPGCQOSA-N (4-butylcyclohexyl) N-[(2S)-4-methyl-1-oxo-1-[[(2S)-1-oxo-3-[(3S)-2-oxopyrrolidin-3-yl]propan-2-yl]amino]pentan-2-yl]carbamate Chemical compound CCCCC1CCC(CC1)OC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C[C@@H]2CCNC2=O)C=O FOLCUFKJHSQMEL-BIXPGCQOSA-N 0.000 description 1
- PNHBRYIAJCYNDA-VQCQRNETSA-N (4r)-6-[2-[2-ethyl-4-(4-fluorophenyl)-6-phenylpyridin-3-yl]ethyl]-4-hydroxyoxan-2-one Chemical compound C([C@H](O)C1)C(=O)OC1CCC=1C(CC)=NC(C=2C=CC=CC=2)=CC=1C1=CC=C(F)C=C1 PNHBRYIAJCYNDA-VQCQRNETSA-N 0.000 description 1
- VIMMECPCYZXUCI-MIMFYIINSA-N (4s,6r)-6-[(1e)-4,4-bis(4-fluorophenyl)-3-(1-methyltetrazol-5-yl)buta-1,3-dienyl]-4-hydroxyoxan-2-one Chemical compound CN1N=NN=C1C(\C=C\[C@@H]1OC(=O)C[C@@H](O)C1)=C(C=1C=CC(F)=CC=1)C1=CC=C(F)C=C1 VIMMECPCYZXUCI-MIMFYIINSA-N 0.000 description 1
- QBTROWHSMGZXCV-RQURQNPSSA-N (6r,7r)-1-[(4s,5r)-4-acetyloxy-5-methyl-3-methylidene-6-phenylhexyl]-4,7-dihydroxy-6-(11-phenoxyundecoxy)-2,8-dioxabicyclo[3.2.1]octane-3,4,5-tricarboxylic acid Chemical compound C([C@@H](C)[C@H](OC(C)=O)C(=C)CCC12[C@@H]([C@@H](OCCCCCCCCCCCOC=3C=CC=CC=3)C(O1)(C(O)=O)C(O)(C(O2)C(O)=O)C(O)=O)O)C1=CC=CC=C1 QBTROWHSMGZXCV-RQURQNPSSA-N 0.000 description 1
- QOLHWXNSCZGWHK-BWBORTOCSA-N (6r,7r)-1-[(4s,5r)-4-acetyloxy-5-methyl-3-methylidene-6-phenylhexyl]-4,7-dihydroxy-6-(11-phenoxyundecylcarbamoyloxy)-2,8-dioxabicyclo[3.2.1]octane-3,4,5-tricarboxylic acid Chemical compound C([C@@H](C)[C@H](OC(C)=O)C(=C)CCC12[C@@H]([C@@H](OC(=O)NCCCCCCCCCCCOC=3C=CC=CC=3)C(O1)(C(O)=O)C(O)(C(O2)C(O)=O)C(O)=O)O)C1=CC=CC=C1 QOLHWXNSCZGWHK-BWBORTOCSA-N 0.000 description 1
- UXKLQDCALAWFIU-VKNDCNMPSA-N (6r,7r)-1-[(4s,5r)-4-acetyloxy-5-methyl-3-methylidene-6-phenylhexyl]-4,7-dihydroxy-6-tetradecoxy-2,8-dioxabicyclo[3.2.1]octane-3,4,5-tricarboxylic acid Chemical compound C([C@@H](C)[C@H](OC(C)=O)C(=C)CCC12[C@H](O)[C@H](C(O2)(C(O)=O)C(O)(C(O1)C(O)=O)C(O)=O)OCCCCCCCCCCCCCC)C1=CC=CC=C1 UXKLQDCALAWFIU-VKNDCNMPSA-N 0.000 description 1
- VGNCBRNRHXEODV-XXVHXNRLSA-N (6r,7r)-1-[(4s,5r)-4-acetyloxy-5-methyl-3-methylidene-6-phenylhexyl]-6-dodecoxy-4,7-dihydroxy-2,8-dioxabicyclo[3.2.1]octane-3,4,5-tricarboxylic acid Chemical compound C([C@@H](C)[C@H](OC(C)=O)C(=C)CCC12[C@H](O)[C@H](C(O2)(C(O)=O)C(O)(C(O1)C(O)=O)C(O)=O)OCCCCCCCCCCCC)C1=CC=CC=C1 VGNCBRNRHXEODV-XXVHXNRLSA-N 0.000 description 1
- IGVKWAAPMVVTFX-BUHFOSPRSA-N (e)-octadec-5-en-7,9-diynoic acid Chemical compound CCCCCCCCC#CC#C\C=C\CCCC(O)=O IGVKWAAPMVVTFX-BUHFOSPRSA-N 0.000 description 1
- OTEKOJQFKOIXMU-UHFFFAOYSA-N 1,4-bis(trichloromethyl)benzene Chemical compound ClC(Cl)(Cl)C1=CC=C(C(Cl)(Cl)Cl)C=C1 OTEKOJQFKOIXMU-UHFFFAOYSA-N 0.000 description 1
- MOQCFMZWVKQBAP-UHFFFAOYSA-N 1-[3,5-bis(trifluoromethyl)benzoyl]-n-(4-chlorophenyl)piperidine-3-carboxamide Chemical compound FC(F)(F)C1=CC(C(F)(F)F)=CC(C(=O)N2CC(CCC2)C(=O)NC=2C=CC(Cl)=CC=2)=C1 MOQCFMZWVKQBAP-UHFFFAOYSA-N 0.000 description 1
- JGMXNNSYEFOBHQ-OWOJBTEDSA-N 2-[(e)-4-morpholin-4-ylbut-2-enyl]-1,1-dioxothieno[3,2-e]thiazine-6-sulfonamide Chemical compound O=S1(=O)C=2SC(S(=O)(=O)N)=CC=2C=CN1C\C=C\CN1CCOCC1 JGMXNNSYEFOBHQ-OWOJBTEDSA-N 0.000 description 1
- AKBHYCHPWZPGAH-UHFFFAOYSA-N 2-[3-[(3-chloro-4-methylphenyl)methoxy]azetidine-1-carbonyl]-7-oxa-5-azaspiro[3.4]octan-6-one Chemical compound CC1=C(Cl)C=C(COC2CN(C2)C(=O)C2CC3(C2)COC(=O)N3)C=C1 AKBHYCHPWZPGAH-UHFFFAOYSA-N 0.000 description 1
- OVDGUTHABMXVMI-UHFFFAOYSA-N 3-nitro-4-(propylamino)benzoic acid Chemical compound CCCNC1=CC=C(C(O)=O)C=C1[N+]([O-])=O OVDGUTHABMXVMI-UHFFFAOYSA-N 0.000 description 1
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- UFMQJYHLIUACCG-UHFFFAOYSA-N 8-nitroindolo[2,1-b]quinazoline-6,12-dione Chemical compound C1=CC=C2C(=O)N3C4=CC=C([N+](=O)[O-])C=C4C(=O)C3=NC2=C1 UFMQJYHLIUACCG-UHFFFAOYSA-N 0.000 description 1
- 102000013455 Amyloid beta-Peptides Human genes 0.000 description 1
- 108010090849 Amyloid beta-Peptides Proteins 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 229940126559 Compound 4e Drugs 0.000 description 1
- 229940125761 Compound 6g Drugs 0.000 description 1
- HSWVJQBEXRKOBZ-QGZVFWFLSA-N FC1=C(OC2CCN(CC2)C=2N=C3C(=NC=2N[C@H]2COCC2)CN(CC3)C(C)=O)C=CC(=C1)F Chemical compound FC1=C(OC2CCN(CC2)C=2N=C3C(=NC=2N[C@H]2COCC2)CN(CC3)C(C)=O)C=CC(=C1)F HSWVJQBEXRKOBZ-QGZVFWFLSA-N 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102000003815 Interleukin-11 Human genes 0.000 description 1
- 108090000177 Interleukin-11 Proteins 0.000 description 1
- TZYWCYJVHRLUCT-VABKMULXSA-N N-benzyloxycarbonyl-L-leucyl-L-leucyl-L-leucinal Chemical compound CC(C)C[C@@H](C=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)OCC1=CC=CC=C1 TZYWCYJVHRLUCT-VABKMULXSA-N 0.000 description 1
- TZCCKCLHNUSAMQ-DUGSHLAESA-N NC(=O)C[C@H](NC(=O)[C@H](CCCNC(=N)N)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCNC(=N)N)NC(=O)[C@H](Cc2ccc(F)cc2)NC(=O)[C@H](Cc3c[nH]c4ccccc34)NC(=O)Cc5cccs5)C(=O)N Chemical compound NC(=O)C[C@H](NC(=O)[C@H](CCCNC(=N)N)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCNC(=N)N)NC(=O)[C@H](Cc2ccc(F)cc2)NC(=O)[C@H](Cc3c[nH]c4ccccc34)NC(=O)Cc5cccs5)C(=O)N TZCCKCLHNUSAMQ-DUGSHLAESA-N 0.000 description 1
- 206010029240 Neuritis Diseases 0.000 description 1
- AKQOBHZKBDHWQI-BZEFIUHZSA-N O[C@H]1C[C@@H](CCC1)N1C(C2(C3=C1N=C(N=C3)NC1=CC=C(C=C1)S(=O)(=O)NC([2H])([2H])[2H])CC2)=O Chemical compound O[C@H]1C[C@@H](CCC1)N1C(C2(C3=C1N=C(N=C3)NC1=CC=C(C=C1)S(=O)(=O)NC([2H])([2H])[2H])CC2)=O AKQOBHZKBDHWQI-BZEFIUHZSA-N 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 229940125907 SJ995973 Drugs 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 102000005747 Transcription Factor RelA Human genes 0.000 description 1
- 108010031154 Transcription Factor RelA Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 150000004075 acetic anhydrides Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 210000001642 activated microglia Anatomy 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 230000006933 amyloid-beta aggregation Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000001713 cholinergic effect Effects 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 229940125872 compound 4d Drugs 0.000 description 1
- 229940126115 compound 4f Drugs 0.000 description 1
- 229940127108 compound 5g Drugs 0.000 description 1
- 229940126136 compound 5i Drugs 0.000 description 1
- 230000002508 compound effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000002027 dichloromethane extract Substances 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229940074383 interleukin-11 Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 206010027175 memory impairment Diseases 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- XZMHJYWMCRQSSI-UHFFFAOYSA-N n-[5-[2-(3-acetylanilino)-1,3-thiazol-4-yl]-4-methyl-1,3-thiazol-2-yl]benzamide Chemical compound CC(=O)C1=CC=CC(NC=2SC=C(N=2)C2=C(N=C(NC(=O)C=3C=CC=CC=3)S2)C)=C1 XZMHJYWMCRQSSI-UHFFFAOYSA-N 0.000 description 1
- 229940094933 n-dodecane Drugs 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 230000003961 neuronal insult Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- FDLYAMZZIXQODN-UHFFFAOYSA-N olaparib Chemical compound FC1=CC=C(CC=2C3=CC=CC=C3C(=O)NN=2)C=C1C(=O)N(CC1)CCN1C(=O)C1CC1 FDLYAMZZIXQODN-UHFFFAOYSA-N 0.000 description 1
- 238000011022 operating instruction Methods 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 238000013149 parallel artificial membrane permeability assay Methods 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000000505 pernicious effect Effects 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 230000009822 protein phosphorylation Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- RWWYLEGWBNMMLJ-YSOARWBDSA-N remdesivir Chemical compound NC1=NC=NN2C1=CC=C2[C@]1([C@@H]([C@@H]([C@H](O1)CO[P@](=O)(OC1=CC=CC=C1)N[C@H](C(=O)OCC(CC)CC)C)O)O)C#N RWWYLEGWBNMMLJ-YSOARWBDSA-N 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
- C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
- C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
- C07D239/32—One oxygen, sulfur or nitrogen atom
- C07D239/42—One nitrogen atom
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
Landscapes
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Neurosurgery (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Psychiatry (AREA)
- Hospice & Palliative Care (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明提供了一种抗神经炎症的二苯乙烯类似物及其制备方法和应用。所述抗神经炎症的二苯乙烯类似物,其结构如下所示;其中,R1为H、卤基、C1~C5饱和直链烷基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O(CH2)mCH3、卤素、羟基、C1~C5饱和直链烷基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。本发明基于分子组装的先导化合物发现方法,通过将嘧啶哌嗪和二苯乙烯两个片段组装,设计得到新骨架分子,并合成了一系列二苯乙烯类似物。本发明提供的化合物具有显著的抗神经炎症活性,并且可以透过血脑屏障,可作为治疗老年痴呆的一种先导化合物。
Description
技术领域
本发明涉及药物设计和药物化学领域,更具体地,涉及一种抗神经炎症的二苯乙烯类似物及其制备方法和应用。
背景技术
阿尔兹海默病(Alzheimer’s disease,AD)俗称老年痴呆,是一种常见的神经系统退行性疾病,伴随着认知功能衰退和记忆损伤。AD的病因复杂,关于该病的发病机制有多种假说:淀粉样级联假说、胆碱能假说、氧化应激假说、Tau蛋白假说等。胞外β-淀粉样蛋白(Aβ)和胞内神经纤维缠结是AD的两个重要标志性病理特征。目前临床上现有治疗AD的方案一般仅能缓解认知功能衰退及缓解认知障碍,无法根本上达到治疗AD的效果。
近年来的研究发现,小胶质细胞主导的神经炎症是AD的一个主要特征,并且神经炎症在AD发病进行中扮演着重要的角色,其会促进并导致AD的发病进程。小胶质细胞是中枢神经系统中的一种巨噬细胞,当大脑受到刺激时,小胶质会启动免疫反应保护大脑免受干扰。但是,在AD中,小胶质细胞处于持续过度激活的状态。Aβ会激活小胶质细胞,被激活的小胶质细胞释放大量的促炎因子和化学因子,如白介素1β(IL-1β),肿瘤坏死因子α(TNF-α),一氧化氮(NO)等,这些因子会对神经元造成损伤并且扩大局部炎症反应。炎症环境也会促进淀粉样斑块和神经纤维缠结的形成,而淀粉样斑块和神经纤维缠结都会使神经元受损。神经元受损会反过来激活小胶质细胞,由此,形成了恶性的神经炎症循环。通过抑制小胶质细胞中促炎因子的形成,抗神经炎症的化合物可以延迟或者终止AD的发病进程,这类化合物的研发受到越来越多的关注。并且在靶向Aβ的化合物接连失败的情况下,抗炎的治疗策略有可能为治疗AD带来新的曙光。
发明内容
本发明的目的之一在于提供新的具有显著抗神经炎症活性的化合物。
本发明的目的之二在于提供所述抗神经炎症化合物的制备方法。
本发明的目的之三在于提供所述化合物在抗神经炎症上的应用。
本发明通过以下技术方案实现上述目的:
本发明提供了一种抗神经炎症的二苯乙烯类似物,所述抗神经炎症的二苯乙烯类似物的结构式如下所示:
其中,R1为H、卤基、C1~C5饱和直链烷基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;
R2为H、O(CH2)mCH3、卤素、羟基、C1~C5饱和直链烷基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;
n为0~3中任意一个整数;m为0~3中任意一个整数。
嘧啶哌嗪是一类抗神经炎症化合物中共有的一个有效活性片段,另外,白藜芦醇作为一种天然产物已被报道具有多种与AD相关的活性,如抗炎、抗氧化、抑制Aβ聚集等,但其生物利用度较差。为得到具有较好的抗神经炎症活性的新骨架分子,基于分子组装的先导化合物发现方法,通过将嘧啶哌嗪和二苯乙烯两个片段组装,设计并合成了一系列化合物。
优选地,R1为H、卤基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O(CH2)mCH3、卤素、羟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。
优选地,R1为H、卤基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O(CH2)mCH3、卤素、羟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。
优选地,R1为H、氟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O(CH2)mCH3、氟基、氯基、羟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。
优选地,其中,R1为H、3-CF3或4-F;R2为H、4-OCH3、2-OCH3、4-Cl、3-Cl、2-Cl、4-F、4-CF3、2-OH或5-OCH3。
最优选地,其中,R1为3-CF3;R2为H。
本发明提供所述的抗神经炎症的二苯乙烯类似物的制备方法,包括如下步骤:将和在碱性条件下进行Perkin反应,然后再与1-(2-嘧啶基)哌嗪缩合得到所述的抗神经炎症的二苯乙烯类似物。
本发明提供的化合物具有显著的抗神经炎症活性,并且具备较好的透过血脑屏障的能力,可作为治疗老年痴呆的一种先导化合物。
进一步地,本发明提供的化合物通过在BV2细胞模型上抗炎活性筛选,包括抑制NO、IL-1β和TNF-α生成,发现化合物8b抗炎活性最好,并对其抗炎机制进行研究,结果表明该化合物作用于MAPK信号通路。并且实验测试发现化合物均可透过血脑屏障。
与现有技术相比,本发明具备如下有益效果:
本发明提供的化合物原料易得,制备简单,且具备显著的抗神经炎症活性,所述的抗神经炎症的二苯乙烯类似物通过作用于MAPK信号通路,并透过血脑屏障发挥效果,在制备预防及治疗老年痴呆药物中具备极大的应用前景。
附图说明
图1为本发明合成的一系列化合物的制备路线图。
图2为化合物8b不影响LPS诱导的NF-κB通路激活。
图3为化合物8b抑制LPS诱导的MAPK通路激活图。
具体实施例
下面结合具体实施例和附图进一步说明本发明的内容,但不应理解为对本发明的限制。在不背离本发明精神和实质的情况下,对本发明方法、步骤或条件所作的简单修改或替换,均属于本发明的范围;若未特别指明,实施例中所用的技术手段为本领域技术人员所熟知的常规手段。
实施例1化合物4~6,8的合成
如图1所示,以苯乙酸类化合物为起始原料,与相应的苯甲醛或肉桂醛在碱性条件下通过Perkin反应生成中间体二苯乙烯酸类化合物(1~3,7),再与1-(2-嘧啶基)哌嗪缩合得到目标化合物(4~6,8)。反应路线如下所示:
具体步骤如下:
1、苯甲醛类化合物或肉桂醛(20mmol,1eq.)与苯乙酸类化合物(20mmol,1eq.)加到20mL乙酸酐中,接着把三乙胺(20mmol,1eq.)缓慢加入,90℃下反应过夜。将2mL水加入到反应液中,继续反应15分钟。然后,26g碳酸钾溶解在150mL水中,将该溶液缓慢加进反应瓶中,60℃下搅拌一个小时。一个小时后,冰浴下加入浓盐酸调节PH为中性或弱酸性。随后,用二氯甲烷萃取,有机层干燥浓缩,乙酸乙酯重结晶得到纯的中间体(1~3,7)。
2、将上述步骤中得到的中间体(1eq.),1-(2-嘧啶基)哌嗪(1.5eq.),EDC.HCl(1eq),三乙胺(1eq),二氯甲烷20mL,分别加入到50mL圆底烧瓶中,室温搅拌,过夜。反应结束后,减压除去有机溶剂,硅胶柱分离(二氯甲烷:乙酸乙酯10:1)得目标产物(4~6,8)。
终产物4~6,8的结构、外观和核磁共振谱图数据如以下所示。
化合物4a:
4a:白色固体(81%).1H NMR(400MHz,CDCl3):δ8.29(2H,t,J=4.7Hz),7.36-7.26(5H,m),7.20-7.12(5H,m),6.78(1H,s),6.51(1H,t,J=4.7Hz),3.69(8H,m).13C NMR(100MHz,CDCl3):δ170.5,161.7,157.8(2×C),137.4,135.6,135.3,130.7,129.5(2×C),128.9(4×C),128.3(2×C),128.3,128.0,110.5,43.7(4×C).ESI-HRMS[M+H]+m/z=371.1859,calcd for C23H22N4O,371.1866.
化合物4b:
4b:白色固体(71%).1H NMR(400MHz,CDCl3):δ8.19(2H,d,J=4.7Hz),7.32-7.26(2H,m),7.25-7.17(3H,m),6.99(2H,d,J=8.7Hz),6.63(3H,m),6.41(1H,t,J=4.7Hz),3.68(3H,s),3.66-3.17(8H,m).13C NMR(100MHz,CDCl3):δ169.7,160.5,158.3,156.7(2×C),134.9,134.2,129.8(2×C),129.5,127.8(4×C),126.9,126.6,112.6(2×C),109.3,54.1,42.5(4×C).ESI-HRMS[M+H]+m/z=401.1965,calcd for C24H24N4O2,401.1972.
化合物4c:
4c:白色固体(63%).1H NMR(400MHz,CDCl3):δ8.33(2H,d,J=4.7Hz),7.62(1H,s),7.56-7.47(2H,m),7.38(1H,t,J=7.8Hz),7.26-7.20(1H,m),7.04(1H,s),6.89(2H,m),6.70(1H,t,J=7.5Hz),6.57-6.50(1H,m),3.82(3H,s),3.82-3.52(8H,m).13C NMR(100MHz,CDCl3):δ170.0,161.5,157.7(2×C),157.6,136.5,134.9,132.1,130.0,129.7,128.9,128.2,125.5,124.5,124.4,123.5,120.2,110.6,110.4,55.4,43.6(4×C).ESI-HRMS[M+H]+m/z=401.1965,calcd for C24H24N4O2,401.1972.
化合物4d:
4d:淡黄色固体(77%).1H NMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.39-7.28(5H,m),7.19-7.13(2H,m),7.06(2H,d,J=8.5Hz),6.74(1H,s),6.52(1H,t,J=4.7Hz),3.83-3.46(8H,m).13C NMR(100MHz,CDCl3):δ170.1,161.5,157.7(2×C),138.1,135.1,133.6,130.7(2×C),129.3,128.9(2×C),128.7(2×C),128.4(4×C),110.4,43.6(4×C).ESI-HRMS[M+H]+m/z=405.1475,calcd for C23H21N4OCl,405.1477.
化合物4e:
4e:白色固体(68%).1H NMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.40-7.30(5H,m),7.21-7.16(1H,m),7.16-7.13(1H,m),7.10(1H,d,J=7.9Hz),7.00(1H,d,J=7.7Hz),6.73(1H,s),6.53(1H,t,J=4.7Hz),3.91-3.54(8H,m).13C NMR(100MHz,CDCl3):δ169.9,161.5,157.7(2×C),138.8,137.0,134.9,134.0,129.3(2×C),129.0,128.9(2×C),128.6(2×C),128.5,127.9,127.4,110.4,43.7(4×C).ESI-HRMS[M+H]+m/z=405.1475,calcd for C23H21N4OCl,405.1477.
化合物4f:
4f:淡黄色固体(64%).1H NMR(400MHz,CDCl3):δ8.31(2H,d,J=4.7Hz),7.41(1H,d,J=7.9Hz),7.31-7.28(2H,m),7.27-7.21(3H,m),7.20-7.14(1H,m),7.00-6.97(2H,m),6.91(1H,s),6.55-6.49(1H,t,J=4.7Hz),3.89-3.57(8H,m).13C NMR(100MHz,CDCl3):δ169.7,161.5,157.7(2×C),138.8,134.4,134.2,134.1,130.9,129.2,128.9,128.7(2×C),128.6(2×C),128.2,127.3,126.4,110.4,46.7,43.9,43.6,41.8.ESI-HRMS[M+H]+m/z=405.1489,calcd for C23H21N4OCl,405.1477.
化合物4g:
4g:白色固体(74%).1H NMR(400MHz,CDCl3):δ8.31(2H,d,J=4.7Hz),7.46(2H,d,J=8.3Hz),7.33(5H,s),7.25(2H,d,J=8.2Hz),6.81(1H,s),6.54(1H,t,J=4.7Hz),3.89-3.49(8H,m).13C NMR(100MHz,CDCl3):δ169.7,161.4,157.7(2×C),139.7,138.8,134.8,129.6(2×C),129.0(2×C),128.8,128.6(3×C),125.1(3×C),125.0,110.5,43.6,43.5,43.5,43.4.ESI-HRMS[M+H]+m/z=439.1744,calcd for C24H21N4OF3,439.1740.
化合物4h:
4h:淡黄色固体(64%).1H NMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.41-7.37(2H,m),7.31-7.26(3H,m),6.96(1H,d,J=8.9Hz),6.78(1H,dd,J=8.9,3.0Hz),6.66(1H,s),6.52(1H,t,J=4.7Hz),6.49(1H,d,J=3.0Hz),3.88-3.51(8H,m),3.43(3H,s).13CNMR(100MHz,CDCl3):δ169.8,161.4,157.7(2×C),156.6,142.7,139.6,134.7,129.2,128.8(2×C),128.7(2×C),128.3,124.5,122.8,115.7,114.4,110.4,55.2,46.4,43.4,41.7,38.9.ESI-MS[M+H]+m/z=417.1,C24H24N4O3.
化合物5a:
5a:淡黄色固体(71%).1H NMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.61(1H,s),7.56-7.50(2H,m),7.41(1H,t,J=7.8Hz),7.23-7.16(3H,m),7.09(2H,dd,J=7.3,2.1Hz),6.88(1H,s),6.54(1H,t,J=4.7Hz),3.72(8H,m).13CNMR(100MHz,CDCl3):δ169.7,161.4,157.7(2×C),136.3,135.5,134.4,132.3,132.0,131.4,129.3(2×C),128.4(3×C),125.7(2×C),124.8(2×C),110.5,43.6(4×C).ESI-HRMS[M+H]+m/z=439.1735,calcdfor C24H21N4OF3,439.1740.
化合物5b:
5b:淡黄色固体(62%).1H NMR(400MHz,CDCl3):δ8.33(2H,d,J=4.7Hz),7.66(1H,s),7.60-7.55(2H,m),7.46(1H,t,J=7.8Hz),7.04(2H,t,J=5.7Hz),6.82(1H,s),6.77-6.73(2H,m),6.55(1H,t,J=4.7Hz),3.80(3H,s),3.66(8H,m).13C NMR(100MHz,CDCl3):δ170.1,161.4,159.7,157.7(2×C),136.8,133.4,132.4,131.9,130.8(2×C),129.3,126.8,125.7,125.7,124.7,124.6,113.8(2×C),110.5,55.2,43.6(4×C).ESI-HRMS[M+H]+m/z=469.1847,calcd for C25H23N4O2F3,469.1846.
化合物5c:
5c:淡黄色固体(70%).1H NMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.60(1H,s),7.53-7.46(2H,m),7.35(1H,t,J=7.8Hz),7.24-7.18(1H,m),7.01(1H,s),6.89-6.83(2H,m),6.68(1H,t,J=7.5Hz),6.52(1H,t,J=4.7Hz),3.87-3.52(11H,overlay).13C NMR(100MHz,CDCl3):δ170.0,161.5,157.7(2×C),157.6,136.5,134.9,132.1,130.0,129.7,128.9,128.2,125.5,125.5,124.5,124.4,123.5,120.2,110.6,110.4,55.4,43.6(4×C).ESI-HRMS[M+H]+m/z=469.1845,calcd for C25H23N4O2F3,469.1846.
化合物5d:
5d:淡黄色固体(79%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.63(1H,s),7.59(1H,d,J=7.7Hz),7.53(1H,d,J=7.8Hz),7.46(1H,t,J=7.7Hz),7.23-7.17(2H,m),7.04(2H,d,J=8.4Hz),6.82(1H,s),6.55(1H,t,J=4.7Hz),3.72(8H,m).13C NMR(100MHz,CDCl3):δ169.4,161.4,157.7(2×C),136.3,136.0,134.2,132.9,132.2,130.6,130.5(2×C),129.4,128.6(2×C),125.6,125.5,125.1,125.1,110.6,43.6,43.6,43.5,43.5.ESI-HRMS[M+H]+m/z=473.1358,calcdfor C24H20N4OF3Cl,473.1351.
化合物5e:
5e:淡黄色固体(76%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.63-7.52(3H,m),7.46(1H,t,J=7.7Hz),7.22(1H,d,J=8.0Hz),7.16-7.10(2H,m),6.95(1H,d,J=7.7Hz),6.80(1H,s),6.55(1H,t,J=4.7Hz),3.73(8H,m).13CNMR(100MHz,CDCl3):δ168.2,160.4,156.7(2×C),136.0,135.3,134.7,133.3,131.1,129.3,128.6,128.4,128.2,127.4,126.3,124.6,124.5,124.2,124.1,109.6,42.7,42.6,42.5,42.4.ESI-HRMS[M+H]+m/z=473.1355,calcd for C24H20N4OF3Cl,473.1351.
化合物5f:
5f:淡黄色固体(61%).1H NMR(400MHz,CDCl3):δ8.33(2H,t,J=4.8Hz),7.58(1H,s),7.51-7.46(2H,m),7.42(1H,dd,J=8.0,0.9Hz),7.36(1H,t,J=7.8Hz),7.19(1H,td,J=7.7,1.4Hz),7.04-6.95(2H,m),6.91(1H,dd,J=7.9,1.3Hz),6.54(1H,t,J=4.7Hz),3.89-3.57(8H,m).13C NMR(100MHz,CDCl3):δ169.0,161.5,157.7(2×C),137.2,135.3,134.1,133.5,132.0,130.6,129.5,129.4,129.1,128.9,126.6,125.5,125.5,124.9,124.9,110.5,43.5(4×C).ESI-HRMS[M+H]+m/z=473.1354,calcd for C24H20N4OF3Cl,473.1351.
化合物5g:
5g:淡黄色固体(61%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.62(1H,s),7.59-7.52(2H,m),7.46(1H,t,J=7.7Hz),7.12-7.05(2H,m),6.95-6.87(2H,m),6.83(1H,s),6.54(1H,t,J=4.7Hz),3.60(8H,m).13C NMR(100MHz,CDCl3):δ169.5,163.7,161.4,157.7(2×C),136.1,135.5,132.2,131.5,131.1,131.0,130.8,130.5,130.4,129.4,125.6,125.0,115.6,115.4,110.5,43.6(4×C).ESI-HRMS[M+H]+m/z=457.1649,calcd for C24H20N4OF4,457.1646.
化合物5h:
5h:淡黄色固体(76%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.60(2H,d,J=8.9Hz),7.55-7.43(4H,m),7.22(2H,d,J=8.3Hz),6.89(1H,s),6.55(1H,t,J=4.7Hz),3.73(8H,m).13C NMR(100MHz,CDCl3):δ169.0,161.4,157.7(2×C),138.1,137.9,135.7,132.1,131.7,131.4,130.2,129.5(4×C)125.5,125.5,125.3,125.3,125.2,110.6,43.6(4×C).ESI-HRMS[M+H]+m/z=507.1618,calcd for C25H20N4OF6,507.1614.
化合物5i:
5i:淡黄色固体(52%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.72(1H,s),7.59-7.51(2H,m),7.40(1H,t,J=7.8Hz),6.98(1H,d,J=8.9Hz),6.81(1H,dd,J=8.9,3.0Hz),6.73(1H,s),6.55(1H,t,J=4.7Hz),6.41(1H,d,J=3.0Hz),3.86-3.53(8H,m),3.46(3H,s).13C NMR(100MHz,CDCl3):δ169.6,169.1,161.4,157.7(2×C),156.9,142.7,138.0,132.3,129.1,128.8,126.2,125.6,125.5,124.9,124.9,123.1,115.9,114.4,110.6,55.3,43.6(4×C).ESI-MS[M+H]+m/z=485.1,calcd for C25H23F3N4O3.
化合物6a:
6a:淡黄色固体(73%).1H NMR(400MHz,CDCl3):δ8.29(2H,d,J=4.7Hz),7.35-7.28(2H,m),7.20-7.10(5H,m),7.02-6.93(2H,m),6.76(1H,s),6.51(1H,t,J=4.7Hz),3.90-3.43(8H,m).13C NMR(100MHz,CDCl3):δ170.2,163.6,161.5,157.7(2×C),136.1,134.9,131.4,130.7,130.7,130.6,129.3(2×C),128.2(2×C),128.0,116.0,115.8,110.5,43.6(4×C).HR-ESI-MS[M+H]+m/z=389.1754,calcd for C23H21N4OF,389.1772.
化合物6b:
6b:淡黄色固体(67%).1H NMR(400MHz,CDCl3):δ8.29(2H,d,J=4.7Hz),7.35-7.31(2H,m),7.01(4H,m),6.74-6.69(3H,m),6.51(1H,t,J=4.7Hz),3.76(3H,s),3.74-3.44(8H,m).13C NMR(100MHz,CDCl3):δ170.5,161.5,159.4,157.7(2×C),134.0,132.4,130.8,130.8,130.7,130.6,130.6,128.7,127.3,116.0,115.8,113.74,113.7,110.5,55.1,43.6(4×C).HR-ESI-MS[M+H]+m/z=419.1861,calcd for C24H23N4O2F,419.1878.
化合物6c:
6c:淡黄色固体(61%).1H NMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.32-7.28(2H,m),7.23-7.17(1H,m),6.97-6.89(4H,m),6.86(1H,d,J=8.3Hz),6.68(1H,t,J=7.4Hz),6.52(1H,t,J=4.7Hz),3.90-3.56(overlap,11H).13C NMR(100MHz,CDCl3):δ169.5,162.4,160.4,160.0,156.7,156.6,134.3,130.5,129.5,129.4,129.1,128.3,125.6,122.9,119.1,114.7,114.5,109.4,109.4,54.4,42.7(4×C).HR-ESI-MS[M+H]+m/z=419.1868,calcd for C24H23N4O2F,419.1878.
化合物6d:
6d:淡黄色固体(76%).1H NMR(400MHz,CDCl3):δ8.31(2H,d,J=4.7Hz),7.32-7.27(2H,m),7.21-7.16(2H,m),7.09-6.98(4H,m),6.73(1H,s),6.54(1H,t,J=4.7Hz),3.86-3.46(8H,m).13C NMR(100MHz,CDCl3):δ169.9,163.7,161.5,157.7(2×C),136.8,133.8,133.4,130.8,130.6,130.6(2×C),130.5,129.4,128.5(2×C),116.2,116.0,110.5,43.6(4×C).HR-ESI-MS[M+H]+m/z=423.1358,calcd for C23H20N4OFCl,423.1382.
化合物6e:
6e:淡黄色固体(65%).1H NMR(400MHz,CDCl3):δ8.30(2H,t,J=4.7Hz),7.32-7.27(2H,m),7.19-7.14(1H,m),7.16-7.08(2H,m),7.06-6.95(3H,m),6.71(1H,s),6.53(1H,t,J=4.7Hz),3.86-3.43(8H,m).13C NMR(100MHz,CDCl3):δ169.7,163.8,161.5,157.7(2×C),137.6,136.8,134.2,130.8,130.6,130.5,129.5,129.2,129.0,128.0,127.3,116.1,115.9,110.5,43.6(4×C).HR-ESI-MS[M+H]+m/z=423.1374,calcd forC23H20N4OFCl,423.1382.
化合物6f:
6f:淡黄色固体(61%).1H NMR(400MHz,CDCl3):δ8.31(2H,d,J=4.7Hz),7.43-7.38(1H,m),7.30-7.25(2H,m),7.19-7.15(1H,m),7.03-6.90(4H,m),6.88(1H,s),6.53(1H,t,J=4.7Hz),3.76(8H,m).13C NMR(100MHz,CDCl3):δ169.5,163.7,161.5,161.2,157.7(2×C),137.6,134.1,134.0,130.8,130.6,130.5,129.3,129.1,127.4,126.5,115.8,115.6,110.5,43.5(4×C).HR-ESI-MS[M+H]+m/z=423.1368,calcd forC23H20N4OFCl,423.1382.
化合物6g:
6g:淡黄色固体(78%).1HNMR(400MHz,CDCl3):δ8.30(2H,d,J=4.7Hz),7.35-7.29(2H,m),7.12-7.06(2H,m),7.01-6.96(2H,m),6.93-6.84(2H,m),6.74(1H,s),6.52(1H,t,J=4.7Hz),3.66(8H,m).13C NMR(100MHz,CDCl3):δ170.0,163.6,161.5,161.1,157.7(2×C),136.1,131.2,131.1,131.0,131.0,130.6,130.5,129.6,116.1,115.9,115.4,115.2,110.5,43.6(4×C).HR-ESI-MS[M+H]+m/z=407.1687,calcd for C23H20N4OF2,407.1678.
化合物6h:
6h:淡黄色固体(71%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.48(2H,d,J=8.0Hz),7.36-7.29(2H,m),7.25(2H,d,J=8.0Hz),7.07-6.99(2H,m),6.80(1H,s),6.55(1H,t,J=4.7Hz),3.86-3.49(8H,m).13C NMR(100MHz,CDCl3):δ169.5,163.9,161.5,157.7(2×C),138.6,138.5,130.7,130.6,130.5,129.5(2×C),128.9,125.3,125.2,125.2,125.2,116.3,116.1,110.6,43.6(4×C).HR-ESI-MS[M+H]+m/z=457.1639,calcdfor C24H20N4OF4,457.1646.
化合物6i:
6i:黄色固体(54%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.77-7.69(1H,m),7.57-7.51(1H,m),7.43-7.35(2H,m),7.02-6.95(3H,m),6.80(1H,dd,J=8.9,3.0Hz),6.64(1H,s),6.54(1H,t,J=4.7Hz),3.72(8H,m),3.50(3H,s).13C NMR(100MHz,CDCl3):δ169.6,167.6,161.4,157.7,157.7,142.7,138.5,130.8,130.7,130.6,129.2,128.7,124.6,122.9,115.8,115.6,115.5,114.6,110.5,55.3,43.8,43.8,43.6,43.4.ESI-MS[M+H]+m/z=435.1,C24H23FN4O3.
化合物8a:
8a:黄色固体(58%).1H NMR(400MHz,CDCl3):δ8.28(2H,t,J=4.7Hz),7.46-7.39(2H,m),7.37-7.33(2H,m),7.30(2H,t,J=7.3Hz),7.26-7.21(1H,m),7.15-7.08(2H,m),7.00(1H,dd,J=15.5,11.1Hz),6.81(1H,d,J=15.6Hz),6.68(1H,d,J=10.8Hz),6.52(1H,t,J=4.7Hz),3.91-3.43(8H,m).13C NMR(100MHz,CDCl3):δ170.1,161.5,161.1,157.7(2×C),137.8,136.6,135.6,131.7,131.7,130.8,130.7,128.7(2×C),128.4,126.9(2×C),123.9,115.9,115.7,110.5,43.6(4×C).HR-ESI-MS[M+H]+m/z=415.1912,calcd forC25H23N4OF,415.1929.
化合物8b:
8b:黄色固体(45%).1H NMR(400MHz,CDCl3):δ8.32(2H,d,J=4.7Hz),7.75(1H,s),7.66(2H,t,J=8.2Hz),7.62-7.55(1H,m),7.42-7.23(5H,m),7.00(1H,dd,J=13.9,12.3Hz),6.88(1H,d,J=15.5Hz),6.76(1H,d,J=10.9Hz),6.54(1H,m),3.54(8H,m).13CNMR(100MHz,CDCl3):δ169.8,161.6,157.8(2×C),139.0,136.6,136.4,135.0,133.1,132.4,129.3,128.9(2×C),128.7,127.0(2×C),125.8,125.6,124.8,124.7,123.4,110.5,43.6(4×C).HR-ESI-MS[M+H]+m/z=465.1888,calcd for C26H23N4OF3,465.1897.
实施例2实施例1所得的28个化合物的抗炎活性测试实验
(1)细胞培养。体外培养BV2细胞(小鼠小胶质细胞)。使用含有10%胎牛血清,链霉素(100微克/毫升),青霉素(100单位/毫升)的DMEM培养基,在37℃,5%二氧化碳浓度条件下进行常规维持培养和传代。
(2)药物干预。取处于对数生长期的细胞,消化,离心,重悬后按每孔100μL,每孔细胞数4×104个接种于96孔板中,培养24小时后吸除原培养液,加入不含血清的DMEM培养基80μL,30分钟后加入不同浓度的目标化合物10μL,最终的浓度为(0.5、1、5、10、25、50μM),空白对照组和LPS单独处理组不加,预处理30分钟后,各组加入LPS(10μg/mL)10μL,即最终浓度为1μg/mL,放入培养箱中继续培养24h后检测。
(3)Griess法检测目标化合物抑制NO实验。按试剂盒说明测定培养液中的NO水平,主要步骤如下:
①按50μL/孔,在96孔板中加入标准品及样品。
②按50μL/孔,在各孔中加入室温Griess试剂I。
③按50μL/孔,在各孔中加入室温Griess试剂II。
④540nm测定吸光度。
(4)ELISA法检测目标化合物抑制IL-1β和TNF-α实验。ELISA Kit是典型的夹心法酶联免疫吸附测定试剂盒(Enzyme Linked-Immuno-Sorbent Assay,ELISA)。预先包被的抗体为单克隆抗体。检测相抗体为多克隆抗体,经生物素(biotin)标记。样品和生物素标记抗体先后加入酶标板孔反应后,用PBS或TBS洗涤多次,去除非特异性吸附物。随后,加入过氧化物酶标记的亲和素进行反应;经过PBS或TBS的彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的IL-1β或TNF-α呈正相关。具体按照试剂盒操作说明进行。主要步骤如下:
①加样品和标准品,37℃反应90分钟。不洗。
②加生物素标记抗体,37℃反应60分钟。TBS洗涤3次。
③加ABC,37℃反应30分钟。TBS洗涤5次。
④TMB 37℃反应20-25分钟。
⑤加入TMB终止液,读数。
结果处理。按照公式:(FL-FC)/(FL-F0)×100%,其中,FL是LPS处理组的OD值,FC是化合物处理组的OD值,F0是未处理细胞组的OD值,计算各化合物在不同浓度下对NO,IL-1β或TNF-α生成的相对抑制率。以相对抑制率为纵坐标,化合物浓度为横坐标绘制各化合物对NO,IL-1β或TNF-α生成的抑制曲线图,并计算各化合物的半数抑制剂量(IC50)以评价各化合物对NO,IL-1β或TNF-α生成的抑制活性。如表1所示,化合物4~6,8的IC50。说明化合物5h、6h、8a、8b能有效抑制NO,IL-1β和TNF-α生成。其中化合物8b抗炎活性最好,故对其进行抗炎机制研究。
表1化合物4~6,8抑制NO,IL-1β和TNF-α生成的IC50和透过BBB能力
实施例3实施例1所得的28个化合物的PAMPA-BBB实验
1.实验原理。PAMPA是一种可用于测定化合物通过(和保留于)浓缩的带负电荷的磷脂双分子层屏障的通透性的方法。它能快速的提供关于被动转运的信息,是一个优异的细胞模型替代物。2003年,Di等人将猪脑组织提取物代替磷脂,将其溶解于十二烷中(2%wt/vol),模拟血脑屏障。
2.试剂准备
(1)配制2%的猪脑提取物(Porcine brain lipid,PBL):称取20mg猪脑提取物加入1mL的正十二烷,使其充分溶解。使用前配制。
(2)配制50mM的PBS:称取1.36g K2HPO4于200mL超纯水中,KOH调pH至7.4。
(3)配制5mg/mL对照药物储备液:称取5mg对照药物加入1mL DMSO溶解,-20℃保存。
(4)配制100μg/mL待测样品液:取储备液20μL于1.5mL的EP管中,加入980μl缓冲液(pH 7.4PBS:乙醇=70:30)。
3.操作步骤
(1)小心吸取4μL 2%的猪脑提取液加入作为给药池的96孔板疏水膜上。
(2)迅速吸取200μL待测样品液于96孔板中作为给药池,接收池加入300uL缓冲液(pH 7.4PBS:乙醇:DMSO=68:30:2)。
(3)小心的将给药池平放于接收池上,并使膜与接收液充分接触。
(4)室温静止10小时,小心移去给药池。用多功能酶标仪测试接收池内化合物在其最大吸收峰处的OD值。
(5)吸取200μL待测样品液于300μL缓冲液(pH 7.4 PBS:乙醇:DMSO=68:30:2)中混匀,作为理论平衡溶液,测其化合物在最大吸收峰处的OD值。
(6)以含2%的DMSO缓冲液作为空白对照。
(7)根据公式计算出Pe值
Pe=-Vd×Va/[(Vd+Va)A×t]×ln(1-OD接收池内待测样品/OD理论平衡溶液待测样品)
Vd为给药池体积,Vd为接收池体积,A为渗透面积,t为渗透时间。当Pe>4.7时既表示化合物具有较好的透过血脑屏障能力。如下所示:
实施例1所得的28个化合物的Pe值如表1所示,化合物的Pe均大于4.7,说明化合物均可透过血脑屏障,并且比白藜芦醇透过血脑屏障的能力提高了。
实施例4实施例1所得的化合物8b的抗炎机制研究
MAPK和NF-κB是两条主要的炎症通路,为探究化合物8b的抗炎机制,分别对两条通路的关键蛋白进行Western blot检测。
1、细胞处理。取处于对数生长期的细胞,消化,离心,重悬后按每孔2mL,
每孔细胞数1×106个接种于6孔板中,培养24h后吸除原培养液,加入不同浓度的8b孵育30分钟,然后加入LPS(1μg/ml)作用16个小时。培养箱中取出六孔板置于冰上,将原培养液吸除干净,用冷的PBS清洗两遍,加入RIPA裂解液裂解细胞,超声破碎,然后转移至EP管中,13000r·min-1离心15min。离心完后,取上清至新的EP管中,取20μL进行蛋白定量。
2、Western blot检测。SDS-PAGE凝胶电泳分离蛋白,然后将蛋白转移至PVDF膜上,转膜结束后将膜取出,先在TBST溶液中清洗一遍,然后放入TBST配制的5%脱脂奶粉封闭液中,室温封闭1~2h。封闭结束后,TBST清洗一遍,加入到由封闭液配制的相应的一抗溶液中,4℃孵育12h。一抗孵育结束后,将膜用TBST清洗3次,每次3min。然后将膜放入到由封闭液配制的相应的二抗溶液中,室温孵育2h。二抗孵育结束后,将膜用TBST清洗3次,每次3min待用。最后,进行发光显影。
3、Western blot结果。化合物8b对MAPK和NF-κB两条炎症通路的作用如图2和图3所示。LPS会激活NF-κB通路,从而使IκBα磷酸化和NF-κB p65从胞核转移到胞浆。但与LPS处理组相比,加入化合物8b对IκBα的磷酸化和p65的转移并无影响,故化合物8b不作用于NF-κB通路。LPS也会激活MAPK通路,MAPK通路有三个关键蛋白:JNK、ERK和p38 MAPK。LPS通过激活MAPK通路,使蛋白磷酸化,与LPS处理组相比,加入化合物8b对JNK、ERK和p38 MAPK这三个蛋白的磷酸化均有抑制作用,并且抑制效果与化合物的浓度呈现剂量依赖关系,故化合物8b抑制了LPS诱导的MAPK信号通路的激活。通过8b对MAPK和NF-κB两条炎症通路的Westernblot结果分析说明:化合物8b通过抑制MAPK信号通路起到抗炎的作用。
上述实验结果表明本发明所述化合物具有显著的抗神经炎症的作用,通过抑制MAPK信号通路起到抗炎的作用,并且能够透过血脑屏障,可作用治疗AD的先导分子。
Claims (9)
1.一种抗神经炎症的二苯乙烯类似物,其特征在于,所述抗神经炎症的二苯乙烯类似物的结构式如下所示:
其中,R1为H、卤基、C1~C5饱和直链烷基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;
R2为H、O (CH2)mCH3、卤素、羟基、C1~C5饱和直链烷基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;
n为0~3中任意一个整数;m为0~3中任意一个整数。
2.根据权利要求1所述的抗神经炎症的二苯乙烯类似物,其特征在于,R1为H、卤基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O (CH2)mCH3、卤素、羟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。
3.根据权利要求1所述的抗神经炎症的二苯乙烯类似物,其特征在于,R1为H、卤基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O (CH2)mCH3、卤素、羟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。
4.根据权利要求1所述的抗神经炎症的二苯乙烯类似物,其特征在于,R1为H、氟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基;R2为H、O (CH2)mCH3、氟基、氯基、羟基、被1个或多个卤素原子取代的C1~C3饱和直链烷基。
5.根据权利要求1所述的抗神经炎症的二苯乙烯类似物,其特征在于,其中,R1为H、3-CF3或4-F;R2为H、4-OCH3、2-OCH3、4-Cl、3-Cl、2-Cl、4-F、4-CF3、2-OH或5-OCH3。
6.根据权利要求5所述的抗神经炎症的二苯乙烯类似物,其特征在于,其中,R1为3-CF3;R2为H。
7.一种权利要求1所述的抗神经炎症的二苯乙烯类似物的制备方法,其特征在于,包括如下步骤:将和在碱性条件下进行Perkin反应,然后再与1-(2-嘧啶基)哌嗪缩合得到所述的抗神经炎症的二苯乙烯类似物。
8.权利要求1至6中任一所述的抗神经炎症的二苯乙烯类似物在制备抗阿尔兹海默病药物中的应用。
9.根据权利要求8所述的应用,其特征在于,所述药物包括药学上可接受的盐。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810134754.4A CN108358853B (zh) | 2018-02-09 | 2018-02-09 | 一种抗神经炎症的二苯乙烯类似物及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810134754.4A CN108358853B (zh) | 2018-02-09 | 2018-02-09 | 一种抗神经炎症的二苯乙烯类似物及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN108358853A true CN108358853A (zh) | 2018-08-03 |
CN108358853B CN108358853B (zh) | 2021-06-25 |
Family
ID=63005596
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810134754.4A Active CN108358853B (zh) | 2018-02-09 | 2018-02-09 | 一种抗神经炎症的二苯乙烯类似物及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108358853B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110496117A (zh) * | 2019-09-30 | 2019-11-26 | 中国科学院微生物研究所 | 一种小分子化合物在抗非洲猪瘟病毒感染中的新应用 |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1445211A (zh) * | 2002-03-20 | 2003-10-01 | 中国医学科学院药物研究所 | 新的番荔枝酰胺衍生物及其制法和其药物组合物与用途 |
CN101142174A (zh) * | 2005-03-19 | 2008-03-12 | 株式会社Amorepacific | 作为香草素受体拮抗剂的新化合物,其异构体或其药用盐;和包含其的药物组合物 |
CN101851211A (zh) * | 2009-03-31 | 2010-10-06 | 中国科学院广州生物医药与健康研究院 | 一种新型哌嗪酰胺类化合物 |
WO2011002067A1 (ja) * | 2009-07-02 | 2011-01-06 | 武田薬品工業株式会社 | 複素環化合物およびその用途 |
WO2012117421A1 (en) * | 2011-03-02 | 2012-09-07 | Orchid Research Laboratories Ltd | Histone deacetylase inhibitors |
CN104693121A (zh) * | 2014-11-14 | 2015-06-10 | 中山大学 | 一类具有抗老年痴呆活性的吡唑类化合物、制备方法与应用 |
CN107365274A (zh) * | 2017-07-19 | 2017-11-21 | 广州医大科技发展有限公司 | 嘧啶哌嗪酰胺类化合物及其应用 |
-
2018
- 2018-02-09 CN CN201810134754.4A patent/CN108358853B/zh active Active
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1445211A (zh) * | 2002-03-20 | 2003-10-01 | 中国医学科学院药物研究所 | 新的番荔枝酰胺衍生物及其制法和其药物组合物与用途 |
CN101142174A (zh) * | 2005-03-19 | 2008-03-12 | 株式会社Amorepacific | 作为香草素受体拮抗剂的新化合物,其异构体或其药用盐;和包含其的药物组合物 |
CN101851211A (zh) * | 2009-03-31 | 2010-10-06 | 中国科学院广州生物医药与健康研究院 | 一种新型哌嗪酰胺类化合物 |
WO2011002067A1 (ja) * | 2009-07-02 | 2011-01-06 | 武田薬品工業株式会社 | 複素環化合物およびその用途 |
WO2012117421A1 (en) * | 2011-03-02 | 2012-09-07 | Orchid Research Laboratories Ltd | Histone deacetylase inhibitors |
CN104693121A (zh) * | 2014-11-14 | 2015-06-10 | 中山大学 | 一类具有抗老年痴呆活性的吡唑类化合物、制备方法与应用 |
CN107365274A (zh) * | 2017-07-19 | 2017-11-21 | 广州医大科技发展有限公司 | 嘧啶哌嗪酰胺类化合物及其应用 |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110496117A (zh) * | 2019-09-30 | 2019-11-26 | 中国科学院微生物研究所 | 一种小分子化合物在抗非洲猪瘟病毒感染中的新应用 |
CN110496117B (zh) * | 2019-09-30 | 2022-09-06 | 中国科学院微生物研究所 | 一种小分子化合物在抗非洲猪瘟病毒感染中的新应用 |
Also Published As
Publication number | Publication date |
---|---|
CN108358853B (zh) | 2021-06-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Fantini et al. | Synthetic soluble analogs of galactosylceramide (GalCer) bind to the V3 domain of HIV-1 gp120 and inhibit HIV-1-induced fusion and entry | |
Nour et al. | The antiprotozoal activity of sixteen asteraceae species native to Sudan and bioactivity-guided isolation of xanthanolides from Xanthium brasilicum | |
Wang et al. | Synthesis, biological evaluation and molecular docking studies of aminochalcone derivatives as potential anticancer agents by targeting tubulin colchicine binding site | |
Manzo et al. | Structure and synthesis of a unique isonitrile lipid isolated from the marine mollusk Actinocyclus papillatus | |
CN108358853A (zh) | 一种抗神经炎症的二苯乙烯类似物及其制备方法和应用 | |
CN109574910A (zh) | 一种咔唑衍生物mcab及其制备方法和应用 | |
Latif et al. | Protoflavone-chalcone hybrids exhibit enhanced antitumor action through modulating redox balance, depolarizing the mitochondrial membrane, and inhibiting ATR-dependent signaling | |
Li et al. | ‘Click chemistry’synthesis of novel natural product-like caged xanthones bearing a 1, 2, 3-triazole moiety with improved druglike properties as orally active antitumor agents | |
Novotortsev et al. | Dispirooxindoles based on 2-selenoxo-imidazolidin-4-ones: Synthesis, cytotoxicity and ROS generation ability | |
Das et al. | Non-vesicular lipid transport machinery in Entamoeba histolytica | |
Salga et al. | Synthesis, characterization, acetylcholinesterase inhibition, molecular modeling and antioxidant activities of some novel schiff bases derived from 1-(2-ketoiminoethyl) piperazines | |
Wang et al. | Novel perbutyrylated glucose derivatives of (–)-epigallocatechin-3-gallate inhibit cancer cells proliferation by decreasing phosphorylation of the EGFR: Synthesis, cytotoxicity, and molecular docking | |
Yang et al. | Development of 3‐phenyl‐N‐(2‐(3‐phenylureido) ethyl)‐thiophene‐2‐sulfonamide compounds as inhibitors of antiapoptotic Bcl‐2 family proteins | |
CN103181918A (zh) | 脂肪酸类化合物在制备预防和治疗肝癌药物中的应用 | |
WO2021252488A1 (en) | Inhibitors of nek7 kinase | |
Hong et al. | Synthesis and biological evaluation of analogues of butyrolactone I as PTP1B inhibitors | |
Zuccolini et al. | IK Channel-independent effects of clotrimazole and senicapoc on cancer cells viability and migration | |
Rivera-Chávez et al. | Hydroxy-neo-Clerodanes and 5, 10-seco-neo-Clerodanes from Salvia decora | |
Lee et al. | A new sulfonic acid derivative,(Z)-4-methylundeca-1, 9-diene-6-sulfonic acid, isolated from the cold water sea urchin inhibits inflammatory responses through JNK/p38 MAPK and NF-κB inactivation in RAW 264.7 | |
Hong et al. | Anti-inflammatory activity of Cajanin, an isoflavonoid derivative isolated from Canavalia lineata pods | |
CN106432025A (zh) | 金刚烷磺酰胺类化合物及其制备与应用 | |
CN110511214A (zh) | 二胺基取代芳杂环类化合物及其制备方法和应用 | |
Grudzień et al. | The Effect of Xanthohumol Derivatives on Apoptosis Induction in Canine Lymphoma and Leukemia Cell Lines | |
Petrova et al. | α-Glucosidase Inhibitors Based on Oleanolic Acid for the Treatment of Immunometabolic Disorders | |
CN108822089A (zh) | 2-芳基咪唑烷类化合物、制备方法及其作为Hsp90抑制剂在抗肿瘤上的应用 |
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 |