CN115616216A - 抑制或阻断faah与nlrp3之间相互作用的制剂的用途 - Google Patents
抑制或阻断faah与nlrp3之间相互作用的制剂的用途 Download PDFInfo
- Publication number
- CN115616216A CN115616216A CN202110800301.2A CN202110800301A CN115616216A CN 115616216 A CN115616216 A CN 115616216A CN 202110800301 A CN202110800301 A CN 202110800301A CN 115616216 A CN115616216 A CN 115616216A
- Authority
- CN
- China
- Prior art keywords
- nlrp3
- faah
- interaction
- protein
- agent
- 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.)
- Pending
Links
- 102000000874 Pyrin Domain-Containing 3 Protein NLR Family Human genes 0.000 title claims abstract description 186
- 108010001946 Pyrin Domain-Containing 3 Protein NLR Family Proteins 0.000 title claims abstract description 186
- 230000003993 interaction Effects 0.000 title claims abstract description 49
- 102100027297 Fatty acid 2-hydroxylase Human genes 0.000 title claims abstract 10
- 101000937693 Homo sapiens Fatty acid 2-hydroxylase Proteins 0.000 title claims abstract 10
- 101000918494 Homo sapiens Fatty-acid amide hydrolase 1 Proteins 0.000 title claims abstract 10
- 108091008099 NLRP3 inflammasome Proteins 0.000 claims abstract description 27
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 20
- 230000035772 mutation Effects 0.000 claims abstract description 18
- 230000000903 blocking effect Effects 0.000 claims abstract description 8
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 8
- 201000010099 disease Diseases 0.000 claims description 19
- 239000003795 chemical substances by application Substances 0.000 claims description 17
- WUADCCWRTIWANL-UHFFFAOYSA-N biochanin A Chemical compound C1=CC(OC)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O WUADCCWRTIWANL-UHFFFAOYSA-N 0.000 claims description 16
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 11
- 239000003814 drug Substances 0.000 claims description 9
- 208000011580 syndromic disease Diseases 0.000 claims description 9
- MLGPKWUKOQAAGI-UHFFFAOYSA-N N-benzylhexadecanamide Chemical compound CCCCCCCCCCCCCCCC(=O)NCC1=CC=CC=C1 MLGPKWUKOQAAGI-UHFFFAOYSA-N 0.000 claims description 6
- HWEFRGHMJOCVPQ-UHFFFAOYSA-N C1CC(CCC1)NC(O)=O.C(N)(=O)C=1C=C(C=CC1)C1=CC=CC=C1 Chemical compound C1CC(CCC1)NC(O)=O.C(N)(=O)C=1C=C(C=CC1)C1=CC=CC=C1 HWEFRGHMJOCVPQ-UHFFFAOYSA-N 0.000 claims description 5
- YWGYNGCRVZLMCS-UHFFFAOYSA-N n-(4-chloropyridin-3-yl)-4-[(2,2-difluoro-1,3-benzodioxol-5-yl)methyl]piperazine-1-carboxamide Chemical compound C1=C2OC(F)(F)OC2=CC=C1CN(CC1)CCN1C(=O)NC1=CN=CC=C1Cl YWGYNGCRVZLMCS-UHFFFAOYSA-N 0.000 claims description 5
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
- 239000005906 Imidacloprid Substances 0.000 claims description 3
- 230000001684 chronic effect Effects 0.000 claims description 3
- 229940056881 imidacloprid Drugs 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 230000000737 periodic effect Effects 0.000 claims description 3
- 229920001184 polypeptide Polymers 0.000 claims description 3
- 208000024827 Alzheimer disease Diseases 0.000 claims description 2
- 201000001320 Atherosclerosis Diseases 0.000 claims description 2
- 208000022715 Autoinflammatory syndrome Diseases 0.000 claims description 2
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 claims description 2
- 208000004232 Enteritis Diseases 0.000 claims description 2
- 201000005569 Gout Diseases 0.000 claims description 2
- 206010034010 Parkinsonism Diseases 0.000 claims description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 claims description 2
- 208000006673 asthma Diseases 0.000 claims description 2
- 208000006454 hepatitis Diseases 0.000 claims description 2
- 231100000283 hepatitis Toxicity 0.000 claims description 2
- YWTYJOPNNQFBPC-UHFFFAOYSA-N imidacloprid Chemical compound [O-][N+](=O)\N=C1/NCCN1CC1=CC=C(Cl)N=C1 YWTYJOPNNQFBPC-UHFFFAOYSA-N 0.000 claims description 2
- 201000006417 multiple sclerosis Diseases 0.000 claims description 2
- 201000008383 nephritis Diseases 0.000 claims description 2
- 210000000653 nervous system Anatomy 0.000 claims description 2
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 230000004913 activation Effects 0.000 abstract description 26
- 238000002360 preparation method Methods 0.000 abstract description 6
- 206010061218 Inflammation Diseases 0.000 abstract description 3
- 230000004054 inflammatory process Effects 0.000 abstract description 3
- 208000027866 inflammatory disease Diseases 0.000 abstract description 2
- 102100029111 Fatty-acid amide hydrolase 1 Human genes 0.000 description 141
- 108010046094 fatty-acid amide hydrolase Proteins 0.000 description 141
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 42
- ROFVXGGUISEHAM-UHFFFAOYSA-N URB597 Chemical compound NC(=O)C1=CC=CC(C=2C=C(OC(=O)NC3CCCCC3)C=CC=2)=C1 ROFVXGGUISEHAM-UHFFFAOYSA-N 0.000 description 36
- 239000002158 endotoxin Substances 0.000 description 36
- 229920006008 lipopolysaccharide Polymers 0.000 description 36
- 230000015556 catabolic process Effects 0.000 description 29
- 238000006731 degradation reaction Methods 0.000 description 29
- 210000004027 cell Anatomy 0.000 description 28
- 238000004458 analytical method Methods 0.000 description 26
- 238000003119 immunoblot Methods 0.000 description 25
- 230000002829 reductive effect Effects 0.000 description 25
- 102000003777 Interleukin-1 beta Human genes 0.000 description 22
- 108090000193 Interleukin-1 beta Proteins 0.000 description 22
- 241000699670 Mus sp. Species 0.000 description 19
- 230000000694 effects Effects 0.000 description 19
- DANUORFCFTYTSZ-UHFFFAOYSA-N epinigericin Natural products O1C2(C(CC(C)(O2)C2OC(C)(CC2)C2C(CC(O2)C2C(CC(C)C(O)(CO)O2)C)C)C)C(C)C(OC)CC1CC1CCC(C)C(C(C)C(O)=O)O1 DANUORFCFTYTSZ-UHFFFAOYSA-N 0.000 description 15
- 230000002757 inflammatory effect Effects 0.000 description 15
- DANUORFCFTYTSZ-BIBFWWMMSA-N nigericin Chemical compound C([C@@H]1C[C@H]([C@H]([C@]2([C@@H](C[C@](C)(O2)C2O[C@@](C)(CC2)C2[C@H](CC(O2)[C@@H]2[C@H](C[C@@H](C)[C@](O)(CO)O2)C)C)C)O1)C)OC)[C@H]1CC[C@H](C)C([C@@H](C)C(O)=O)O1 DANUORFCFTYTSZ-BIBFWWMMSA-N 0.000 description 15
- 230000004900 autophagic degradation Effects 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 108090000623 proteins and genes Proteins 0.000 description 14
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 13
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 13
- 210000003470 mitochondria Anatomy 0.000 description 13
- 239000013592 cell lysate Substances 0.000 description 12
- 208000022993 cryopyrin-associated periodic syndrome Diseases 0.000 description 12
- 210000004379 membrane Anatomy 0.000 description 12
- 239000012528 membrane Substances 0.000 description 12
- 238000010384 proximity ligation assay Methods 0.000 description 12
- 230000001960 triggered effect Effects 0.000 description 12
- 108090000426 Caspase-1 Proteins 0.000 description 11
- 230000017854 proteolysis Effects 0.000 description 9
- 102100035904 Caspase-1 Human genes 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 8
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 8
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 8
- 230000000638 stimulation Effects 0.000 description 8
- 101001024605 Homo sapiens Next to BRCA1 gene 1 protein Proteins 0.000 description 7
- 101710173438 Late L2 mu core protein Proteins 0.000 description 7
- 102100037001 Next to BRCA1 gene 1 protein Human genes 0.000 description 7
- 101710188315 Protein X Proteins 0.000 description 7
- 230000001965 increasing effect Effects 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 230000034512 ubiquitination Effects 0.000 description 7
- 238000010798 ubiquitination Methods 0.000 description 7
- WKSHMJCYWFOADB-UHFFFAOYSA-N (4-morpholin-4-yl-1,2,5-thiadiazol-3-yl) N-cyclooctyl-N-methylcarbamate Chemical compound C1(CCCCCCC1)N(C(OC1=NSN=C1N1CCOCC1)=O)C WKSHMJCYWFOADB-UHFFFAOYSA-N 0.000 description 6
- 208000035690 Familial cold urticaria Diseases 0.000 description 6
- 206010064570 familial cold autoinflammatory syndrome Diseases 0.000 description 6
- BATCTBJIJJEPHM-UHFFFAOYSA-N n-pyridazin-3-yl-4-[[3-[5-(trifluoromethyl)pyridin-2-yl]oxyphenyl]methylidene]piperidine-1-carboxamide Chemical compound N1=CC(C(F)(F)F)=CC=C1OC1=CC=CC(C=C2CCN(CC2)C(=O)NC=2N=NC=CC=2)=C1 BATCTBJIJJEPHM-UHFFFAOYSA-N 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 5
- 108010034143 Inflammasomes Proteins 0.000 description 5
- -1 N-Benzylpalmitamide (N-Benzylpalmitamide) Chemical compound 0.000 description 5
- JLVTVCRXFMLUIF-UHFFFAOYSA-N O-Desmorpholinopropyl Gefitinib Chemical compound C=12C=C(O)C(OC)=CC2=NC=NC=1NC1=CC=C(F)C(Cl)=C1 JLVTVCRXFMLUIF-UHFFFAOYSA-N 0.000 description 5
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 description 5
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 description 5
- 239000003940 fatty acid amidase inhibitor Substances 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 210000004185 liver Anatomy 0.000 description 5
- 230000037452 priming Effects 0.000 description 5
- SEGYOKHGGFKMCX-UHFFFAOYSA-N 4-[bis(1,3-benzodioxol-5-yl)-hydroxymethyl]-1-piperidinecarboxylic acid (4-nitrophenyl) ester Chemical compound C=1C=C2OCOC2=CC=1C(C=1C=C2OCOC2=CC=1)(O)C(CC1)CCN1C(=O)OC1=CC=C([N+]([O-])=O)C=C1 SEGYOKHGGFKMCX-UHFFFAOYSA-N 0.000 description 4
- 102100029647 Apoptosis-associated speck-like protein containing a CARD Human genes 0.000 description 4
- 102100029115 Fumarylacetoacetase Human genes 0.000 description 4
- GZNIYOXWFCDBBJ-UHFFFAOYSA-N N,N-dimethyl-5-[(4-phenylphenyl)methyl]-1-tetrazolecarboxamide Chemical compound CN(C)C(=O)N1N=NN=C1CC1=CC=C(C=2C=CC=CC=2)C=C1 GZNIYOXWFCDBBJ-UHFFFAOYSA-N 0.000 description 4
- NBOJHRYUGLRASX-UHFFFAOYSA-N N-(3-pyridinyl)-4-[[3-[[5-(trifluoromethyl)-2-pyridinyl]oxy]phenyl]methyl]-1-piperidinecarboxamide Chemical compound N1=CC(C(F)(F)F)=CC=C1OC1=CC=CC(CC2CCN(CC2)C(=O)NC=2C=NC=CC=2)=C1 NBOJHRYUGLRASX-UHFFFAOYSA-N 0.000 description 4
- 150000001200 N-acyl ethanolamides Chemical class 0.000 description 4
- HSYCMGWPPRTNKH-UHFFFAOYSA-N [2-(methylamino)-2-oxoethyl] n-[2-[1-(6-methylpyridin-2-yl)piperidin-4-yl]ethyl]carbamate Chemical compound C1CC(CCNC(=O)OCC(=O)NC)CCN1C1=CC=CC(C)=N1 HSYCMGWPPRTNKH-UHFFFAOYSA-N 0.000 description 4
- 238000011278 co-treatment Methods 0.000 description 4
- 229960003638 dopamine Drugs 0.000 description 4
- 239000002621 endocannabinoid Substances 0.000 description 4
- 238000003197 gene knockdown Methods 0.000 description 4
- 238000001114 immunoprecipitation Methods 0.000 description 4
- QNYRAEKLMNDRFY-UHFFFAOYSA-N jzl195 Chemical compound C1=CC([N+](=O)[O-])=CC=C1OC(=O)N1CCN(CC=2C=C(OC=3C=CC=CC=3)C=CC=2)CC1 QNYRAEKLMNDRFY-UHFFFAOYSA-N 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- DOWVMJFBDGWVML-UHFFFAOYSA-N n-cyclohexyl-n-methyl-4-(1-oxidopyridin-1-ium-3-yl)imidazole-1-carboxamide Chemical compound C1=NC(C=2C=[N+]([O-])C=CC=2)=CN1C(=O)N(C)C1CCCCC1 DOWVMJFBDGWVML-UHFFFAOYSA-N 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 230000002269 spontaneous effect Effects 0.000 description 4
- 201000003274 CINCA syndrome Diseases 0.000 description 3
- 102100036361 Calcium-binding and coiled-coil domain-containing protein 2 Human genes 0.000 description 3
- 102100029968 Calreticulin Human genes 0.000 description 3
- 101000714579 Homo sapiens Calcium-binding and coiled-coil domain-containing protein 2 Proteins 0.000 description 3
- 101000793651 Homo sapiens Calreticulin Proteins 0.000 description 3
- 102000000589 Interleukin-1 Human genes 0.000 description 3
- 108010002352 Interleukin-1 Proteins 0.000 description 3
- 108020004459 Small interfering RNA Proteins 0.000 description 3
- 108010087999 Steryl-Sulfatase Proteins 0.000 description 3
- 239000012822 autophagy inhibitor Substances 0.000 description 3
- 230000030570 cellular localization Effects 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 3
- 230000001086 cytosolic effect Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 238000013507 mapping Methods 0.000 description 3
- 230000002438 mitochondrial effect Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 230000000087 stabilizing effect Effects 0.000 description 3
- QDLHCMPXEPAAMD-QAIWCSMKSA-N wortmannin Chemical compound C1([C@]2(C)C3=C(C4=O)OC=C3C(=O)O[C@@H]2COC)=C4[C@@H]2CCC(=O)[C@@]2(C)C[C@H]1OC(C)=O QDLHCMPXEPAAMD-QAIWCSMKSA-N 0.000 description 3
- QDLHCMPXEPAAMD-UHFFFAOYSA-N wortmannin Natural products COCC1OC(=O)C2=COC(C3=O)=C2C1(C)C1=C3C2CCC(=O)C2(C)CC1OC(C)=O QDLHCMPXEPAAMD-UHFFFAOYSA-N 0.000 description 3
- VCMMYRWIEZCYDK-PDBXOOCHSA-N (9z,12z,15z)-n-benzyloctadeca-9,12,15-trienamide Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(=O)NCC1=CC=CC=C1 VCMMYRWIEZCYDK-PDBXOOCHSA-N 0.000 description 2
- HUUSXLKCTQDPGL-UHFFFAOYSA-N 1-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)-3-[4-(2-hydroxypropan-2-yl)furan-2-yl]sulfonylurea Chemical compound CC(C)(O)C1=COC(S(=O)(=O)NC(=O)NC=2C=3CCCC=3C=C3CCCC3=2)=C1 HUUSXLKCTQDPGL-UHFFFAOYSA-N 0.000 description 2
- 102000016614 Autophagy-Related Protein 5 Human genes 0.000 description 2
- 108010092776 Autophagy-Related Protein 5 Proteins 0.000 description 2
- 206010064568 Chronic infantile neurological cutaneous and articular syndrome Diseases 0.000 description 2
- 102100023147 E3 ubiquitin-protein ligase MARCHF7 Human genes 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 101150008770 FAAH gene Proteins 0.000 description 2
- 101000978673 Homo sapiens E3 ubiquitin-protein ligase MARCHF7 Proteins 0.000 description 2
- 108010052285 Membrane Proteins Proteins 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 2
- 102000005398 Monoacylglycerol Lipase Human genes 0.000 description 2
- 108020002334 Monoacylglycerol lipase Proteins 0.000 description 2
- 101150061038 NLRP3 gene Proteins 0.000 description 2
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 2
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 2
- 229940079156 Proteasome inhibitor Drugs 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 238000004873 anchoring Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 210000004957 autophagosome Anatomy 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- IVUMCTKHWDRRMH-UHFFFAOYSA-N carprofen Chemical compound C1=CC(Cl)=C[C]2C3=CC=C(C(C(O)=O)C)C=C3N=C21 IVUMCTKHWDRRMH-UHFFFAOYSA-N 0.000 description 2
- 229960003184 carprofen Drugs 0.000 description 2
- 230000009089 cytolysis Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- ZGSPNIOCEDOHGS-UHFFFAOYSA-L disodium [3-[2,3-di(octadeca-9,12-dienoyloxy)propoxy-oxidophosphoryl]oxy-2-hydroxypropyl] 2,3-di(octadeca-9,12-dienoyloxy)propyl phosphate Chemical compound [Na+].[Na+].CCCCCC=CCC=CCCCCCCCC(=O)OCC(OC(=O)CCCCCCCC=CCC=CCCCCC)COP([O-])(=O)OCC(O)COP([O-])(=O)OCC(OC(=O)CCCCCCCC=CCC=CCCCCC)COC(=O)CCCCCCCC=CCC=CCCCCC ZGSPNIOCEDOHGS-UHFFFAOYSA-L 0.000 description 2
- 238000010494 dissociation reaction Methods 0.000 description 2
- 230000005593 dissociations Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 238000011813 knockout mouse model Methods 0.000 description 2
- 210000001865 kupffer cell Anatomy 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000003207 proteasome inhibitor Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 230000009885 systemic effect Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- GFARQYQBWJLZMW-JYFOCSDGSA-N (3e,5e)-3,5-bis[(4-nitrophenyl)methylidene]-1-prop-2-enoylpiperidin-4-one Chemical compound C1=CC([N+](=O)[O-])=CC=C1\C=C(/CN(C\1)C(=O)C=C)C(=O)C/1=C/C1=CC=C([N+]([O-])=O)C=C1 GFARQYQBWJLZMW-JYFOCSDGSA-N 0.000 description 1
- VCMMYRWIEZCYDK-UHFFFAOYSA-N (9Z,12Z,15Z)-N-benzyloctadeca-9,12,15-trienamide Natural products CCC=CCC=CCC=CCCCCCCCC(=O)NCC1=CC=CC=C1 VCMMYRWIEZCYDK-UHFFFAOYSA-N 0.000 description 1
- ARGIPZKQJGFSGQ-LCYFTJDESA-N (Z)-PYR-41 Chemical compound C1=CC(C(=O)OCC)=CC=C1N(NC\1=O)C(=O)C/1=C\C1=CC=C([N+]([O-])=O)O1 ARGIPZKQJGFSGQ-LCYFTJDESA-N 0.000 description 1
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 1
- CVRREEFNKPDIOA-UHFFFAOYSA-N 2-cyclohexyl-N-methyl-4-(1-oxido-2H-pyridin-1-ium-1-yl)imidazole-1-carboxamide Chemical compound CNC(=O)N1C=C([N+]2([O-])C=CC=CC2)N=C1C1CCCCC1 CVRREEFNKPDIOA-UHFFFAOYSA-N 0.000 description 1
- 108060000255 AIM2 Proteins 0.000 description 1
- 108091006112 ATPases Proteins 0.000 description 1
- 102000057290 Adenosine Triphosphatases Human genes 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 108700023418 Amidases Proteins 0.000 description 1
- IASNWHAGGYTEKX-IUCAKERBSA-N Arg-Arg-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(O)=O IASNWHAGGYTEKX-IUCAKERBSA-N 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- RGEIIUGXLRATHD-UHFFFAOYSA-N CC(C)(C)OC(N(CCN(CC1=CC(OC2=CC=CC=C2)=CC=C1)C1)C1C(C=C1)=CC=C1[N+]([O-])=O)=O Chemical compound CC(C)(C)OC(N(CCN(CC1=CC(OC2=CC=CC=C2)=CC=C1)C1)C1C(C=C1)=CC=C1[N+]([O-])=O)=O RGEIIUGXLRATHD-UHFFFAOYSA-N 0.000 description 1
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- HKQYGTCOTHHOMP-UHFFFAOYSA-N Formononetol Natural products C1=CC(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O HKQYGTCOTHHOMP-UHFFFAOYSA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- CCBIBMKQNXHNIN-ZETCQYMHSA-N Gly-Leu-Gly Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CCBIBMKQNXHNIN-ZETCQYMHSA-N 0.000 description 1
- ABPRMMYHROQBLY-NKWVEPMBSA-N Gly-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)CN)C(=O)O ABPRMMYHROQBLY-NKWVEPMBSA-N 0.000 description 1
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 1
- 101000728679 Homo sapiens Apoptosis-associated speck-like protein containing a CARD Proteins 0.000 description 1
- 101000798951 Homo sapiens Mitochondrial import receptor subunit TOM20 homolog Proteins 0.000 description 1
- 101000979572 Homo sapiens NLR family CARD domain-containing protein 4 Proteins 0.000 description 1
- 101000588545 Homo sapiens Serine/threonine-protein kinase Nek7 Proteins 0.000 description 1
- 108700000788 Human immunodeficiency virus 1 tat peptide (47-57) Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- VZIFYHYNQDIPLI-HJWJTTGWSA-N Ile-Arg-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N VZIFYHYNQDIPLI-HJWJTTGWSA-N 0.000 description 1
- 102100024064 Interferon-inducible protein AIM2 Human genes 0.000 description 1
- ZUKPVRWZDMRIEO-VKHMYHEASA-N L-cysteinylglycine Chemical compound SC[C@H]([NH3+])C(=O)NCC([O-])=O ZUKPVRWZDMRIEO-VKHMYHEASA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000006404 Mitochondrial Proteins Human genes 0.000 description 1
- 108010058682 Mitochondrial Proteins Proteins 0.000 description 1
- 102100034007 Mitochondrial import receptor subunit TOM20 homolog Human genes 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 102100023435 NLR family CARD domain-containing protein 4 Human genes 0.000 description 1
- 241000881234 Nisia Species 0.000 description 1
- 101800001821 Precursor of protein E3/E2 Proteins 0.000 description 1
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 1
- 229940123573 Protein synthesis inhibitor Drugs 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 102100020814 Sequestosome-1 Human genes 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 102100031400 Serine/threonine-protein kinase Nek7 Human genes 0.000 description 1
- 108091027967 Small hairpin RNA Proteins 0.000 description 1
- GKMYGVQDGVYCPC-IUKAMOBKSA-N Thr-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H]([C@@H](C)O)N GKMYGVQDGVYCPC-IUKAMOBKSA-N 0.000 description 1
- 102000002689 Toll-like receptor Human genes 0.000 description 1
- 108020000411 Toll-like receptor Proteins 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- 102000005922 amidase Human genes 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 230000004642 autophagic pathway Effects 0.000 description 1
- 230000007320 autophagy mechanism Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000007248 cellular mechanism Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 108010016616 cysteinylglycine Proteins 0.000 description 1
- 230000002074 deregulated effect Effects 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 230000009504 deubiquitination Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000001085 differential centrifugation Methods 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 230000007937 eating Effects 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 208000024963 hair loss Diseases 0.000 description 1
- 230000003676 hair loss Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 210000004020 intracellular membrane Anatomy 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- GTCAXTIRRLKXRU-UHFFFAOYSA-N methyl carbamate Chemical compound COC(N)=O GTCAXTIRRLKXRU-UHFFFAOYSA-N 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 230000036651 mood Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 230000000626 neurodegenerative effect Effects 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 101800002664 p62 Proteins 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 230000008533 pain sensitivity Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 210000003024 peritoneal macrophage Anatomy 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000026938 proteasomal ubiquitin-dependent protein catabolic process Effects 0.000 description 1
- 239000000007 protein synthesis inhibitor Substances 0.000 description 1
- 230000004063 proteosomal degradation Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 102220001685 rs121908150 Human genes 0.000 description 1
- 102220032518 rs151344629 Human genes 0.000 description 1
- 102220032546 rs180177439 Human genes 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000007958 sleep Effects 0.000 description 1
- 239000004055 small Interfering RNA Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/16—Amides, e.g. hydroxamic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/16—Amides, e.g. hydroxamic acids
- A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
- A61K31/23—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin of acids having a carboxyl group bound to a chain of seven or more carbon atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/27—Esters, e.g. nitroglycerine, selenocyanates of carbamic or thiocarbamic acids, meprobamate, carbachol, neostigmine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4525—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with oxygen as a ring hetero atom
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/50—Pyridazines; Hydrogenated pyridazines
- A61K31/501—Pyridazines; Hydrogenated pyridazines not condensed and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/06—Antigout agents, e.g. antihyperuricemic or uricosuric agents
-
- 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
-
- 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
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
- A61P25/16—Anti-Parkinson drugs
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/34—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 3 only
- C07D311/36—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 3 only not hydrogenated in the hetero ring, e.g. isoflavones
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Urology & Nephrology (AREA)
- Diabetes (AREA)
- Pulmonology (AREA)
- Emergency Medicine (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pain & Pain Management (AREA)
- Psychology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Rheumatology (AREA)
- Physical Education & Sports Medicine (AREA)
- Biochemistry (AREA)
Abstract
本发明提供了抑制或阻断FAAH与NLRP3之间相互作用的制剂的用途,本发明采用抑制或阻断FAAH和NLRP3的相互作用的制剂,使NLRP3蛋白处于不稳定的状态而被降解,达到治疗NLRP3炎症小体激活的相关炎症疾病。本发明采用的制剂还可以有效地治疗NLRP3蛋白突变相关炎症疾病。
Description
技术领域
本发明涉及抑制或阻断FAAH与NLRP3之间相互作用的制剂的用途。
背景技术
炎症小体是多蛋白信号传导平台,可响应外源性或内源性危险信号来驱动炎症反应。在鉴定出的多个不同的炎性小体中,对NLRP3炎性小体的研究最多。除了NLRP3作为传感器外,NLRP3炎性小体还包含一个衔接子(ASC;也称为PYCARD)和一个效应子(胱天蛋白酶1,Caspase 1)。NLRP3炎性小体组装后,胱天蛋白酶1被切割并激活,导致促炎细胞因子IL-1β和IL-18的激活和释放,以及细胞焦亡。NLRP3的NACHT结构域内和周围的突变使NLRP3具有组成型活性,从而引起冷吡啉相关周期性综合征(CAPS),包括家族性冷性自身炎性综合症(FCAS),穆-韦二氏综合征(MWS)和新生儿发作多系统炎性疾病(NOMID,也称为慢性婴儿神经系统皮肤关节综合征或CINCA)。
尽管已经对NLRP3炎性小体了解甚多,但仍有许多悬而未决的问题。问题之一是在炎症小体激活之前是否存在维持NLRP3蛋白稳定性的机制。由于失调的NLRP3炎症小体活性推动了许多炎症、代谢、神经退行性疾病和自身免疫性疾病的发展,因此控制NLRP3炎症小体活性至关重要。根据该原理,细胞中的NLRP3蛋白通常维持在非常低的水平,不足以激活。在称为“启动”的过程中,Toll样受体识别与危险相关的分子模式(DAMPS)或与病原体相关的分子模式(PAMPS),触发大量NLRP3蛋白的合成,但是这些蛋白仍处于没有活性的非组装状态。细胞接到了第二个信号的刺激才会导致NLRP3炎症小体的激活与组装(图1),这种微妙的“两个信号”控制机制可确保仅在绝对必要时才启动NLRP3引发的炎症。一个合理的假说是NLRP3蛋白天生就不稳定,细胞中存在维持其稳定性的机制,但目前对该机制的了解甚微。之前有文献报道处于激活状态的组装好的NLRP3炎性小体复合物会发生自噬降解,但是激活之前未组装的NLRP3蛋白如何维持其稳定性知之甚少。另一个问题是NLRP3蛋白的细胞定位。一般认为,NLRP3蛋白在炎症小体组装之前就存在于线粒体和内质网的膜上,但是将NLRP3锚定于线粒体和内质网的蛋白仍处于争论之中。已发表的文献显示线粒体抗病毒信号(MAVS)、心磷脂(cardiolipin)、线粒体蛋白(mitochondrion)和STING均可将 NLRP3锚定于线粒体或内质网,但这些分子对NLRP3蛋白的锚定作用可能仅限于某些特殊情形,如病毒感染期间,或仅对部分NLRP3蛋白起作用。
脂肪酸酰胺水解酶(FAAH)是内源性大麻素系统中起关键调控作用的的膜蛋白,在细胞中主要定位于内质网和线粒体,通过水解各种脂肪酸酰胺包括内源性大麻素系统的内源性大麻素(AEA)等内源性信号脂质而使其失活,从而调节各种生理过程,例如疼痛、进食、血压、情绪、睡眠等。FAAH是否会影响NLRP3蛋白稳定性或炎症小体激活目前尚未见报道。
发明内容
本发明提供了抑制或阻断FAAH与NLRP3之间相互作用的制剂的用途。
为实现上述目的,所采取的技术方案:抑制或阻断FAAH与NLRP3之间相互作用的制剂在筛选治疗由NLRP3炎症小体引起的疾病的药物中的用途。
本发明提供了抑制或阻断FAAH与NLRP3之间相互作用的制剂在筛选治疗由NLRP3蛋白突变引起的疾病的药物中的用途。
本发明提供了抑制或阻断FAAH与NLRP3之间相互作用的制剂在制备治疗由NLRP3炎症小体引起的疾病的药物中的用途。
本发明提供了抑制或阻断FAAH与NLRP3之间相互作用的制剂在制备治疗由NLRP3蛋白突变引起的疾病的药物中的用途。
优选地,所述由NLRP3炎症小体引起的疾病为Ⅱ型糖尿病、痛风、阿尔兹海默症、动脉粥样硬化、帕金森综合征、多发性硬化症、肌萎缩侧所硬化症、哮喘、慢性阻塞性肺疾病、肾炎、肠炎、肝炎。
优选地,所述由NLRP3蛋白突变引起的疾病为冷吡啉相关周期性综合征。
优选地,所述由NLRP3蛋白突变引起的疾病为家族性寒冷型自身炎症性综合征(FCAS)、穆-韦二氏综合征(MWS)和慢性婴儿神经系统皮肤关节综合征(CINCA)。
优选地,所述抑制或阻断FAAH与NLRP3之间相互作用的制剂包括3'-胺甲酰基联苯-3- 基环己基氨基甲酸酯、鹰嘴豆牙素A、N-苄基棕榈酰胺、N-(4-氯-3-吡啶基)-4-[(2,2-二氟-1,3- 苯并二恶茂-5-基)甲基]-1-哌嗪甲酰胺、4-吗啉基-1,2,5-噻二唑-3-基环辛基(甲基)氨基甲酸酯和N-3-吡啶嗪基-4-[[3-[[5-(三氟甲基)-2-吡啶基]氧基]苯基]亚甲基]-1-哌啶甲酰胺中的至少一种。
优选地,所述抑制或阻断FAAH与NLRP3之间相互作用的制剂包括3'-胺甲酰基联苯-3-基环己基氨基甲酸酯。
优选地,所述抑制或阻断FAAH与NLRP3之间相互作用的制剂包括氨基酸序列如SEQID NO:1所示的多肽。
有益效果:
本发明采用阻断FAAH和NLRP3之间的相互作用的制剂,使NLRP3蛋白处于不稳定的状态而被降解,达到治疗NLRP3炎症小体相关疾病。具体来说,在第一个信号刺激后细胞中产生的NLRP3蛋白处于一个天生不稳定的状态,其稳定性依赖于与膜酶FAAH的相互作用,从而被锚定到线粒体和内质网膜上;阻断NLRP3和FAAH的相互作用可以使NLRP3蛋白从线粒体和内质网膜上解离,然后通过E3泛素蛋白连接酶(CHIP)作用造成NLRP3蛋白的 K48泛素化,最后通过E3泛素蛋白连接酶(NBR1)作用被拉到自噬体降解,导致细胞中的 NLRP3蛋白水平显著减低,细胞接收第二个刺激信号后无法组装成有充分活性的炎症小体,从而达到治疗NLRP3炎症小体相关疾病的效果。另外,不同于目前已知的许多通过抑制 NLRP3蛋白内源性ATPase活性而起作用的药物分子(仅对非突变NLRP3蛋白引发的疾病有效),本发明采用的抑制或阻断FAAH与NLRP3之间相互作用的制剂对突变和非突变NLRP3 蛋白所引发的疾病均有效。
附图说明
图1显示巨噬细胞中NLRP3在非刺激情形下基本上不表达,第一个信号刺激导致高表达但无炎症小体激活能力,只有在第二个信号刺激后被激活产生炎症因子。在有或没有LPS处理的引发的(模拟)BMDM中的NLRP3和GAPDH的免疫印迹分析。
图2显示FAAH敲除导致动物模型中NLRP3炎症小体活化减少,ELISA检测WT,FAAH+/-和FAAH-/-BMDM小鼠腹腔中IL-1β及TNF-a的水平。Serum:血清。
图3显示FAAH敲除导致NLRP3蛋白降解。图中a,在WT,FAAH+/-和FAAH-/-BMDM 中,NLRP3,FAAH和GAPDH的免疫印迹分析。b,在有或没有共同处理CHX的LPS引发的(模拟)WT和FAAH-/-BMDM中,NLRP3和GAPDH的免疫印迹分析。图中c,在有或没有共同处理尼日利亚菌素的LPS引发的(模拟)WT和FAAH-/-BMDM中,NLRP3,FAAH 和GAPDH的免疫印迹分析。
图4显示FAAH敲除导致细胞中NLRP3炎症小体活化减少,ELISA检测WT,FAAH+/- 和FAAH-/-BMDM上清液中IL-1β的水平,这些上清液是LPS刺激的,LPS刺激的随后是Nigericin刺激或LPS刺激后加入3-MA,接着是尼日尔霉素刺激(Nigericin+3MA)。平均值±SEM,n=3。
图5显示FAAH敲除导致的NLRP3蛋白降解依赖于自噬而非蛋白酶体途径,图中a,在有或没有共同处理3MA,CQ或Wortmannin的LPS引发的(模拟)FAAH-/-BMDM中,NLRP3 和GAPDH的免疫印迹分析。b,在已经用扰乱的(scrambke)或ATG5特异性的siRNA转染的LPS引发的FAAH-/-BMDMs(模拟物)中NLRP3和GAPDH的免疫印迹分析。
图6显示FAAH敲除导致NLRP3蛋白的K48泛素化增加,而这个作用为NLRP3蛋白降解所必需,用对照(同种型)或NLRP3特异性抗体免疫沉淀未处理或LPS引发的WT和 FAAH-/-BMDMs的细胞裂解物,然后用K48-Ub,K63-Ub或NLRP3抗体免疫印迹。
图7显示FAAH敲除导致的NLRP3蛋白需要E3连接酶CHIP蛋白,图中a,用NLRP3特异性抗体免疫沉淀从BMDM和FAAH-/-BMDMs中提取的蛋白质,然后用NLRP3,NBR1和 CHIP抗体免疫印迹。b,用CHIP特异性siRNA转染的LPS引发的FAAH-/-BMDM中NLRP3, CHIP和GAPDH的免疫印迹分析。
图8显示FAAH敲除导致的NLRP3蛋白降解需要选择性受体蛋白NBR1蛋白,用NBR1特异性siRNA转染的LPS引发的FAAH-/-BMDM中的NLRP3,NBR1和GAPDH进行免疫印迹分析。输入(input)是指未沉淀的细胞裂解液。
图9显示了FAAH与NLRP3蛋白相互作用并使其稳定,图中a,用同种型或NLRP3特异性抗体免疫沉淀LPS引发的BMDMs的细胞裂解液,然后对NLRP3和FAAH进行免疫印迹分析。b,用FAAH特异性抗体免疫沉淀未处理的或LPS引发的(模拟)BMDMs的细胞裂解物,然后对NLRP3和FAAH进行免疫印迹分析。c,在LPS引发的WT,FAAH+/-和 FAAH-/-BMDM中的NLRP3-FAAH相互作用的邻近结扎测定法(PLA);比例尺,20μm。
图10显示FAAH与NLRP3相互作用的结构域Mapping分析,并由此设计出AS-1短肽。图a,b在HEK293T细胞中共转染带有Flag标签的NLRP3构建体和具有HA标签的FAAH构建体。用抗-Flag抗体免疫沉淀的细胞裂解物中HA和Flag蛋白的免疫印迹分析。
图11显示细胞中突变NLRP3与FAAH相互作用。图中a,在HEK293T细胞中共转染带有Flag标签的野生型和三个突变的NLRP3构建体和具有HA标签的FAAH构建体。用抗 Flag抗体免疫沉淀的细胞裂解液中HA和Flag蛋白的免疫印迹分析。b,用同种型或NLRP3 特异性抗体免疫沉淀LPS引发的NLRP3-R258W敲入BMDMs,然后对NLRP3-R258W和 FAAH进行免疫印迹分析。c,在LPS刺激的NLRP3-R258W敲入BMDM中, NLRP3-R258W-FAAH相互作用的PLA;比例尺,20μm。
图12显示FAAH敲除导致突变NLRP3蛋白降解。在WT,FAAH+/-R258W和 FAAH-/-R258W BMDM中的NLRP3和GAPDH的免疫印迹分析。
图13显示FAAH敲除导致动物模型中突变NLRP3炎症小体活化减少,CAPS疾病症状减轻。a,WT,NLRP3-R258W,FAAH+/-R258W和FAAH-/-R258WBMDM小鼠在12周龄时的照片。b,WT,NLRP3-R258W,FAAH+/-R258W和FAAH-/-R258WBMDM小鼠的腋窝淋巴结,脾脏和肝脏。
图14显示FAAH敲除导致细胞中突变NLRP3炎症小体活化减少。a,在LPS刺激3小时后,对WT,NLRP3-R258W,FAAH+/-R258W和FAAH-/-R258WBMDM上清液中的IL-1 β进行ELISA。平均值±SEM,n=3,**p<0.01。b,在源自WT,NLRP3-R258W,FAAH+/- R258W和R258WFAAH-/-R258W小鼠的LPS刺激的BMDM中的培养上清液(SN)中IL-1β和裂解的caspase-1(p20)以及细胞裂解液(裂解)中的Pro-Caspase-1和Pro-IL-1β的免疫印迹分析。lysis表示沉淀。
图15显示FAAH敲除导致NLRP3蛋白从线粒体及内质网膜上解离,图中a,在LPS诱导的WT和FAAH-/-BMDMs中,从细胞溶解产物的细胞溶质和膜组分分得到的NLRP3,CRT, TOM20和GAPDH的免疫印迹分析。b,用抗NLRP3抗体免疫沉淀WT和FAAH-/-BMDMs 的细胞裂解物,然后对NLRP3,CRT,TOM20进行免疫印迹分析。细胞未刺激(对照)或 LPS刺激3小时。c和d,在LPS引发的WT和FAAH-/-BMDM中,NLRP3-TOM20相互作用的PLA(c)和NLRP3-CRT相互作用(d);比例尺,20μm。
图16显示在FAAH正常表达的细胞中突变NLRP3与线粒体及内质网膜相互作用。a和b,在LPS引发的WT,FAAH+/-R258W和FAAH-/-R258W BMDM中,NLRP3-TOM20相互作用的PLA(a)和NLRP3-CRT相互作用(b);比例尺,20μm。
图17不同FAAH抑制剂对NLRP3蛋白的降解作用。采用JZL195、URB597、LY2183240、PF3845、Biochanin A、N-Benzyllinolenamide、Carprofen、BIA10-2474、FAAH-IN-2、 N-Benzylpalmitamide、1-monomyristin、JNJ-42165279、JZL-184、JZP-430、PF-04457845、SA47 这14种FAAH抑制剂进行测试,其中URB597、Biochanin A、N-Benzylpalmitamide、JNJ-42165279、JZP-430、PF-04457845这6种具有显著活性,这是由在40uM的浓度下将NLRP3蛋白水平降低30%以上的能力所定义的。
图18显示URB597导致NLRP3蛋白与FAAH分离.在LPS引发的WT BMDM中,有或没有URB597处理,NLRP3-FAAH相互作用的PLA比例尺,20μm。
图19显示URB597导致细胞中NLRP3炎症小体活化减少.a,培养上清液(SN)中IL-1β和裂解的caspase-1(p20)以及细胞裂解液(裂解)中的Pro-Caspase-1和Pro-IL-1β的免疫印迹分析。将BMDM用LPS刺激3个小时,然后再用指定浓度的URB597处理3个小时,然后用Nigericin激发30分钟。b,用LPS刺激3小时,然后再用指定浓度的URB597处理3个小时,然后用Nigericin激发30分钟的BMDM上清液中IL-1β的ELISA,平均值±SEM,n= 3,***p<0.001。
图20显示URB597导致的NLRP3蛋白降解通过自噬途径实现。图中,在有或没有共同处理URB597,3MA,CQ或Wortmannin的LPS引发的WT BMDM中,NLRP3和GAPDH的免疫印迹分析。
图21显示URB597在小鼠BMDM细胞中导致突变NLRP3蛋白降解.对用LPS刺激的NLRP3-R258W BMDM中的NLRP3-R258W和GAPDH进行免疫印迹分析,然后再用所示剂量的URB597处理另外3小时。
图22示URB597在CAPS病人单核细胞中导致NLRP3蛋白与FAAH分离。来自患者1 的PBMC中NLRP3-FAAH相互作用的PLA。将细胞用LPS刺激3小时,然后再用40μM URB597处理3小时。
图23显示URB597在CAPS病人单核细胞中导致NLRP3炎症小体活化减少。a,从患者1和她的正常母亲分离的PBMC上清液中IL-1β的ELISA。用LPS刺激细胞3小时,然后再用40μMURB597或10μM MCC950处理3小时。平均值±SEM,n=3,***p<0.001。NS,不显著。b和c,分别将来自患者2(b)和3(c)的PBMC用LPS刺激3小时,然后再用 40μM URB597处理3小时。N2是患者2的正常母亲,而N3是无关的正常男孩。平均值±SEM, n=3,***p<0.001。
图24显示AS-1肽导致NLRP3蛋白与FAAH分离。图中,在有或没有AS-1短肽处理的LPS引发的WT BMDM中,NLRP3-FAAH相互作用的PLA。
图25显示AS-1肽导致NLRP3蛋白降解。图中,在有或没有不同时间和浓度AS-1短肽处理的LPS引发的WT BMDM中,NLRP3和GAPDH的免疫印迹分析。
图26显示AS-1肽导致细胞中NLRP3炎症小体活化减少。培养上清液(SN)中IL-1β和裂解的caspase-1(p20)以及细胞裂解液(裂解)中的Pro-Caspase-1和Pro-IL-1β的免疫印迹分析。将BMDM用LPS刺激3个小时,然后再用不同浓度的AS-1短肽处理3个小时,然后用Nigericin激发30分钟。
具体实施方式
为更好的说明本发明的目的、技术方案和优点,下面将结合具体实施例对本发明作进一步说明。
3'-胺甲酰基联苯-3-基环己基氨基甲酸酯(URB597)的结构式如下式(Ⅰ):
4-硝基苯基4-(3-苯氧基苄基)哌嗪-1-甲酸叔丁酯(JZL195)的结构式如下式(Ⅱ):
5-([1,1'-联苯]-4-甲基)-N,N-二甲基-1H-四氮唑-1-甲酰胺(LY2183240)的结构式如下式(Ⅲ):
N-3-吡啶-4-[[3-[[5-(三氟甲基)-2-吡啶]氧基]苯基]甲基]-1-哌啶羧酰胺(PF-3845)的结构式如下式(Ⅳ):
鹰嘴豆牙素A(Biochanin A)的结构式如下式(Ⅴ):
N-苄基-(9Z,12Z,15Z)-十八碳三烯酰胺(N-Benzyllinolenamide)的结构式如下式(Ⅵ):
卡洛芬(Carprofen)的结构式如下式(Ⅶ):
3-(1-(环己基(甲基氨基甲酰基)-1H-咪唑-4-基)吡啶1-氧化物(BIA10-2474)的结构式如下式(Ⅷ):
4-(3-氯-4-氟苯氨基)-7-甲氧基喹唑啉-6-醇(FAAH-IN-2)的结构式如下式(Ⅸ):
N-苄基棕榈酰胺(N-Benzylpalmitamide)的结构式如下式(Ⅹ):
1-肉豆蔻酸单甘油酯(1-monomyristin)的结构式如下式(Ⅺ):
N-(4-氯-3-吡啶基)-4-[(2,2-二氟-1,3-苯并二恶茂-5-基)甲基]-1-哌嗪甲酰胺(JNJ-42165279) 的结构式如下式(Ⅻ):
4-[双(1,3-苯并二恶唑-5-基)羟甲基]-1-哌啶甲酸4-硝基苯酯(JZL-184)的结构式如下式(XIII):
4-吗啉基-1,2,5-噻二唑-3-基环辛基(甲基)氨基甲酸酯(JZP-430, 4-Morpholino-1,2,5-thiadiazol-3-yl cyclooctyl(methyl)carbamate)的结构式如下式(XIV):
N-3-吡啶嗪基-4-[[3-[[5-(三氟甲基)-2-吡啶基]氧基]苯基]亚甲基]-1-哌啶甲酰胺 (PF-04457845)的结构式如下式(XV):
2-(甲基氨基)-2-氧乙基{2-[1-(6-甲基-2-吡啶基)-4-哌啶基]乙基}氨基甲酸酯(SA47) 的结构式如下式(XVI):
实施例1
1、FAAH的丧失导致NLRP3降解
为了评估FAAH在NLRP3炎症小体激活中的潜在作用,我们使用shRNA敲低了小鼠骨髓衍生的巨噬细胞(BMDM)中的FAAH,这导致脂多糖(LPS)引发后,由尼日利亚菌素触发的裂解的半胱天冬酶1和IL-1β的分泌显着减少,表明FAAH可能影响NLRP3炎性体的激活。为了系统地研究FAAH丢失的影响,我们通过CRISPR-Cas9技术删除FAAH基因的外显子15至外显子2来创建FAAH基因敲除小鼠(FAAH ko-1)。我们通过基因组PCR和基因组测序验证了敲除状态。FAAH-/-小鼠的FAAH酶活性降低,并且在AEA攻击下,在尾部浸入和热板测试中均表现出增强的低温反应和降低的疼痛敏感性。FAAH-/-小鼠也表现出低的繁殖能力,而FAAH+/-小鼠则正常繁殖。与源自野生型小鼠的BMDM相比,携带FAAH 杂合性(FAAH+/-)和纯合性(FAAH-/-)损失的BMDM中由LPS引发后的尼日利亚霉素触发的IL-1β释放分别降低了约50%和70%。在ATP和MSU攻击后,FAAH-/-BMDM中也观察到IL-1β分泌的显着减少。此外,FAAH的丧失也减少了IL-18的释放和焦细胞的死亡。与这些结果一致,我们还观察到在FAAH-/-和FAAH-/-小鼠中全身性LPS施用后,血清IL-1β水平分别降低了约45%和70%(图2)。相反,在全身性LPS给药后,FAAH敲除并没有增加血清TNF-α水平。FAAH的丢失对NLRP3 mRNA水平没有影响,但NLRP3蛋白减少,因为在FAAH+/-和FAAH-/-中观察到NLRP3蛋白水平分别降低了约45%和65%(图3a)。此外,在蛋白质合成抑制剂环己酰亚胺的存在下,FAAH的丧失加速了NLRP3的还原(图 3b),指出蛋白质降解增强是蛋白质水平降低的根本原因。总体而言,这些结果表明,FAAH 的缺失通过诱导NLRP3降解而抑制了NLRP3炎性体的活性。
值得注意的是,虽然我们观察到FAAH+/-细胞中的FAAH蛋白减少了约50%,但在FAAH-/-骨髓细胞以及衍生自骨髓的巨噬细胞(BMDM)中,这一结果是可以预期的,我们使用三种不同的市售抗体从FAAH-/-小鼠身上连续检测到一条蛋白带(Protein X),该蛋白带在Western印迹上的位置与FAAH相同。在FAAH-/-小鼠中,心脏中也检测到了蛋白X,但在我们检查过的其他组织(包括脑和肝)中却不存在,它们在野生型小鼠中表达了高水平的FAAH。由另一家公司通过类似但略有不同的策略创建的第二只FAAH基因敲除小鼠 (FAAH ko-2)在BMDM中也表现出Protein X,但在主要组织中却没有包括脑,肝和肾。从 FAAH-/-小鼠分离的Kupffer细胞不表达蛋白X,在LPS启动后未显示可检测水平的NLRP3 蛋白(图3c),并且尼日尔霉素攻击后IL-1β释放减少了近90%(图4),表明FAAH丢失对Kupffer细胞中NLRP3的影响要比BMDM中的影响更大。值得注意的是,已在人类和许多其他物种中发现了第二个FAAH基因,但在小鼠和大鼠中却未发现。仍在研究蛋白质X的身份及其在NLRP3稳定性和激活中的可能作用。
2、NLRP3通过选择性自噬降解
为了探究由FAAH缺失引起的NLRP3降解的分子机制,我们评估了蛋白酶体和自噬的潜在作用,这是两个负责蛋白质降解的细胞机制。自噬抑制剂3-MA,渥曼青霉素和CQ以及ATG5的敲低均增加了FAAH-/-BMDM中NLRP3的蛋白水平,表明自噬对NLRP3降解的减弱作用(图5a和5b)。相反,蛋白酶体抑制剂MG132,PS341和b-AP15都进一步降低了这些细胞中的NLRP3水平(图5c),表明蛋白酶体抑制后自噬降解增强。这些结果表明,FAAH 的丧失会诱导自噬而不是蛋白酶体降解NLRP3。由于FAAH-/-BMDM中的NLRP3降解表现出高特异性,而未观察到NLRC4,AIM2,ASC和NEK7的水平降低,我们推测该降解是通过选择性自噬进行的,这是通常通过特定自噬受体将泛素化的货物递送至自噬体来完成。支持该假设的是,泛素化抑制剂PYR41降低,而去泛素化抑制剂PR619升高,FAAH-/-BMDM 中的NLRP3降解。此外,与FAAH+/+BMDMs相比,NLRP3在FAAH-/-BMDMs中表现出增强的K48泛素化,但没有K63泛素化(图6)。据报道,多巴胺可诱导巨噬细胞中NLRP3 的泛素化依赖性自噬降解,许多已知的E3连接酶中的两种MARCH7和CHIP在多巴胺处理后与NLRP3相关(但只有MARCH7负责多巴胺诱导的NLRP3降解,结果是我们能够复制的)。我们发现,在FAAH-/-BMDM中,CHIP(而不是MARCH7)与NLRP3绑定(图7a)。此外,CHIP的敲低减少了因FAAH丧失而引起的NLRP3降解(图7b)。为了鉴定所涉及的自噬受体,我们评估了p62,NBR1和NDP52,这是三种可能的候选药物。在FAAH-/-BMDM 中,发现NBR1和NDP52(而非p62)显示出与NLRP3的相互作用增加(图7a)。但是,只有NBR1的敲低,而不是NDP52的敲低,减少了NLRP3的降解(图8)。因此,FAAH 的丧失通过作为E3连接酶的CHIP和作为自噬受体的NBR1介导的选择性自噬诱导了NLRP3 的降解。
3、FAAH与NLRP3蛋白相互作用并使其稳定
为了进一步了解FAAH的丧失如何导致NLRP3降解,我们搜索了与NLRP3相互作用的蛋白质。出乎意料的是,我们发现LRP引发的BMDM中NLRP3与FAAH相互作用,如co-IP 分析所示(图9a,9b)。作为支持,当在HEK293T细胞中共表达时,带有FLAG标签的NLRP3 与带有HA标签的FAAH共同定位。。邻近连接测定法(PLA)进一步揭示了野生型BMDM 中NLRP3-FAAH相互作用很强,并且在FAAH+/-BMDM中这种相互作用显著减弱,而在 FAAH-/-BMDM中则几乎不存在这种相互作用(图9c)。该结果表明,蛋白X不与NLRP3 相互作用,或者,FAAH抗体能够在免疫印迹中识别蛋白X,但在PLA条件下不能识别。在小鼠巨噬细胞系J774A.1中还发现NLRP3与FAAH相关。NLRP3与FAAH之间的相互作用具有高度选择性,因为FAAH不会与其他与炎症小体相关的蛋白(包括ASC,胱天蛋白酶 1,NEK7,NLRC4和AIM2)相互作用,相反,NLRP3不会与单酰基甘油脂肪酶(MAGL) (内源性大麻素系统中的水解酶)相互作用。为了进一步了解NLRP3-FAAH相互作用,我们进行了作图研究。跨越aa 91和aa 710之间区域的NLRP3的NACHT结构域对于与FAAH的相互作用既是必需的又是足够的。进一步的映射揭示了aa 494和aa710之间的区域足以进行 NLRP3与FAAH的相互作用。我们还确定了FAAH的aa 150和aa 265之间的区域对于与 NLRP3的相互作用至关重要,因为FAAH的aa 1至265片段与全长NLRP3相互作用,而aa 1至150的aa片段没有相互作用(图10a,图10b)。有趣的是,该区域构成了大多数酰胺酶签名(AS)序列(FAAH的134-257位残基),这是在AS家族的一大批酶中发现的大约130 个氨基酸的连续序列,其中大多数是细菌和真菌原产地。
以上结果有力地表明,NLRP3具有内在的不稳定性,并通过与FAAH的相互作用得以稳定。为了加强这一结论,我们通过慢病毒表达系统将FAAH重新引入了FAAH-/-BMDM中。这些细胞中FAAH的过度表达“拯救”了FAAH稳定NLRP3的能力,导致LPS引发后NLRP3 的水平更高,并且相应地,尼日尔菌素攻击后导致的IL-1β释放也高于在FAAH+/+BMDM 中观察到的水平。相反,FAAH-S241A的过度表达,其在关键催化亲核细胞Ser241残基39 上具有失活突变,并且在293T细胞中表达时表现出与NLRP3结合的能力降低,在稳定小鼠 BMDM中NLRP3或恢复尼日利亚霉素诱导的NLRP3炎症体激活方面无效。这些结果表明, FAAH需要酶活性才能与NLRP3相互作用并使其稳定。但是,我们不能排除以下可能性:受 S241A突变影响的蛋白质构象变化(而不是酶活性的丧失)对于减少NLRP3相互作用更为关键。
4、FAAH还与NLRP3-R258W相互作用并使其稳定
为了评估FAAH是否也与CAPS相关突变体NLRP3相互作用,我们在293T细胞中表达了HA标记的FAAH以及三个FLAG标记的鼠NLRP3突变体之一(NLRP3-R258W, NLRP3-T346M和NLRP3-F521L)。这些分别对应于在FCAS,MWS和NOMID患者中发现的人NLRP3-R260W,NLRP3-T348M和NLRP3-F523L突变。Co-IP分析显示,FAAH结合了所有三种NLRP3突变蛋白(图11a),这表明与FAAH的相互作用可能是CAPS相关的 NLRP3蛋白的共同特征。此外,我们发现NLRP3-R258W在源自NLRP3-R258W敲入小鼠的 BMDM中与FAAH强烈相互作用(图11b和11c)。在由LPS启动的BMDM中,NLRP3-R258W 的水平与野生型NLRP3相当,表明FAAH在这些细胞中具有稳定野生型和突变型NLRP3的相似能力。
为了评估FAAH丢失对突变体NLRP3的影响,我们通过将NLRP3-R258W小鼠与FAAH-/-小鼠杂交来产生R258W FAAH+/-小鼠。在从R258W FAAH+/-小鼠分离的LPS引发的BMDM中,FAAH的杂合丢失使NLRP3-R258W蛋白质水平降低了40%(图12),由尼日利亚霉素引起的IL-1β释放降低了50%。值得注意的是,FAAH的杂合丢失足以显着降低 NLRP3-R258W突变所表现出的炎症表型。与报道的结果一致,NLRP3-R258W小鼠表现出皱纹的外套,脱发,较小的体重和体重以及严重的皮肤发炎,但是FAAH+/-R258W小鼠在所有这些方面均得到了显着改善(代表性照片如图13a所示)。NLRP3-R258W小鼠还显示出腋窝淋巴结,脾脏和肝脏肿大,而从FAAH+/-R258W小鼠切除的这些器官明显较小(但仍明显比WT小鼠大;图13b)。NLRP3-R258W小鼠的组织学研究显示,受累皮肤的表皮和真皮均增厚,并且肝脏出现明显的门脉炎症,但FAAH+/-R258W小鼠几乎没有这些特征。我们目前正在获取FAAH-/-NLRP3-R258W小鼠。并且FAAH敲除导致细胞中突变NLRP3炎症小体活化减少(图14a,图14b)。综上所述,以上结果证明FAAH在稳定CAPS相关的 NLRP3-R258W蛋白中起关键作用。
5、FAAH将NLRP3锚定于线粒体和ER膜
NLRP3和FAAH之间的相互作用促使我们评估FAAH(一种已知与ER和线粒体相关的细胞内膜蛋白)如何影响NLRP3的细胞定位。使用简单的差分离心方案,在去除大细胞碎片后,我们将LPS引发的BMDMs的细胞裂解液分为细胞溶质组分和膜组分。大部分NLRP3 蛋白存在于膜部分中,该部分含有线粒体而不包含ER,这表明线粒体可能比ER更参与NLRP3 的细胞定位。重要的是,FAAH的缺失导致NLRP3蛋白的细胞溶质分数增加,反之则导致膜分数减少(图15a)。与该结果一致,与FAAH+/+BMDM相比,FAAH-/-BMDM中观察到的NLRP3-TOM20相互作用降低,如co-IP和PLA分析(图15b,15c和15d)所示,表明NLRP3 与FAAH丧失后的线粒体。我们还发现,通过使用ER标记CRT,FAAH-/-BMDM中的 NLRP3-ER相互作用也有类似的降低(图15b和15d)。FAAH似乎对于炎性体激活是必不可少的,因为自噬抑制剂3-MA在FAAH-/-BMDM中“免于”降解的NLRP3表现出相当的能力,可以响应尼日尔菌素的激发而引起IL-1β释放。该结果也强烈表明,FAAH损失对NLRP3 炎性体活化的抑制作用是由于NLRP3蛋白降解,而不是FAAH水解的AEA和其他FAA含量增加,因为这些FAA的增加是由FAAH损失引起的。不太可能受到3-MA治疗的影响。同样,由于FAA的增加,FAAH损失引起的NLRP3降解效应也不大可能。在支持方面,AEA 无法引发NLRP3降解。与FAAH与CAPS突变的NLRP3相互作用的发现一致,FAAH还发挥了将NLRP3-R258W锚定到线粒体和ER膜上的作用(图16a,16b)。
6、选择性FAAH抑制剂诱导NLRP3降解
灭活的S241A突变使FAAH在稳定NLRP3方面失活的事实促使我们评估FAAH抑制剂的潜在NLRP3降解活性。采用JZL195、URB597、LY2183240、PF3845、Biochanin A、 N-Benzyllinolenamide、Carprofen、BIA10-2474、FAAH-IN-2、N-Benzylpalmitamide、 1-monomyristin、JNJ-42165279、JZL-184、JZP-430、PF-04457845、SA47这14种FAAH抑制剂进行测试,其中URB597、Biochanin A、N-Benzylpalmitamide、JNJ-42165279、JZP-430、 PF-04457845这6种具有显著活性,这是由在40uM的浓度下将NLRP3蛋白水平降低30%以上的能力所定义的(图17)。但是,在抑制FAAH的报道的IC50与它们诱导NLRP3降解的活性之间没有观察到明显的关系。抑制FAAH酶活性的能力是否影响NLRP3降解尚待进一步研究。JZL195、URB597、LY2183240、PF3845、Biochanin A、N-Benzyllinolenamid、Carprofen、BIA10-2474、FAAH-IN-2、N-Benzylpalmitamide、1-monomyristin、JNJ-42165279、JZL-184、 JZP-430、PF-04457845、SA47均采购于MCE公司。
选择了受试抑制剂中活性最高的化合物URB597进行进一步研究。除小鼠BMDM以外, URB597还剂量依赖性地降低了小鼠腹膜巨噬细胞和我们已经研究的其他两种巨噬细胞细胞系中的NLRP3蛋白水平。此外,URB597以剂量依赖性方式提高了小鼠BMDM中的自噬水平,自噬抑制剂而非蛋白酶体抑制剂显着消除了URB597降低NLRP3的作用(图5a,5b和 5c)。类似于FAAH丢失的情况,URB597处理导致NLRP3的K48泛素化增强,并促进CHIP 和NBR1与NLRP3的结合,表明相同的选择性自噬机制。毫不奇怪,URB597破坏了小鼠 BMDM中的内源性NLRP3-FAAH相互作用(图18)。另一方面,多巴胺并没有破坏 NLRP3-FAAH的相互作用。
7、URB597抑制野生型和CAPS相关的突变体NLRP3炎性体激活
从其引发NLRP3降解的优越能力可以预期,URB597在LPS引发的小鼠中剂量依赖性地抑制了由NLRP3特异性刺激物(包括尼日菌,ATP,MSU和明矾)触发的IL-1β和裂解的半胱天冬酶1的分泌。BMDM(图19a和图19b)。URB597还抑制了由尼日利亚菌素诱导的凋亡。重要的是,3-MA消除了URB597的NLRP3降解作用(图20),能够“拯救”被URB597 抑制的NLRP3炎性体活化。这提供了额外的证据支持以下观点:NLFA3降解后,NLRP3蛋白降解而不是增加了AEA和其他FAA,是造成观察到的NLRP3炎性体抑制的原因。与IL-1β释放相反,URB597对LPS和尼日利亚菌素处理触发的TNF-α释放影响最小。此外,在源自 NLRP3-R258W敲入小鼠的BMDM中,LPS攻击后,URB597还诱导NLRP3-R258W蛋白质降解并减少了炎性体激活(图21)。
为了提供有关URB597是否可用于治疗CAPS患者的初步证明,我们从三名患者中分离了PBMC,其概况显示在表1中。
表1三个病人的血液生化指标
CAPS突变的NLRP3与FAAH的相互作用,以及在URB597治疗后NLRP3与FAAH的解离在从一个2岁女孩患者1中分离出的PBMC中得到证实(图22),该女孩被诊断为FCAS,其中NLRP3基因存在A354T突变。与从她的母亲那里获得的没有CAPS的PBMC相比,来自患者1的PBMC表现出IL-1β的自发释放和LPS触发的释放均增加(图23a)。重要的是, URB597能够减少自发和LPS触发的IL-1β释放,在LPS触发的IL-1β释放中观察到的抑制率约为50%。值得注意的是,MCC950对该患者无效。从另外两名患者中分离出的PBMCs 中也观察到自发性增强和LPS触发的IL-1β释放增强,其中一个3岁的女孩被诊断为具有 T350M突变的FCAS,另一个7个月大的男孩被诊断为在NLRP3基因中具有M408T突变的 CINCA。URB597再次有效地抑制了自发和LPS触发的IL-1β释放,对于2号和3号病人, LPS触发的IL-1β释放分别具有40%和60%的抑制率(图23b和23c)。
构建FAAH丝氨酸241位点附近的上下游20个氨基酸的多肽(TAT-AS1: YGRKKRRQRRRGGSPLGLGTDIGGSIRFPSAFCG),在BMDM细胞中测试AS-1短肽能阻断FAAH与NLRP3的相互作用(图24),从而导致NLRP3被降解(图25),进一步影响NLRP3 炎症小体的活化(图26)。
最后所应当说明的是,以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
序列表
<110> 华南理工大学
<120> 抑制或阻断FAAH与NLRP3之间相互作用的制剂的用途
<130> WK21-YSY-CN1-0269
<141> 2021-07-15
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 34
<212> PRT
<213> 人工序列(Artificial)
<400> 1
Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Gly Gly Ser Pro Leu
1 5 10 15
Gly Leu Gly Thr Asp Ile Gly Gly Ser Ile Arg Phe Pro Ser Ala Phe
20 25 30
Cys Gly
Claims (10)
1.抑制或阻断FAAH与NLRP3之间相互作用的制剂在筛选治疗由NLRP3炎症小体引起的疾病的药物中的用途。
2.抑制或阻断FAAH与NLRP3之间相互作用的制剂在筛选治疗由NLRP3蛋白突变引起的疾病的药物中的用途。
3.抑制或阻断FAAH与NLRP3之间相互作用的制剂在制备治疗由NLRP3炎症小体引起的疾病的药物中的用途。
4.抑制或阻断FAAH与NLRP3之间相互作用的制剂在制备治疗由NLRP3蛋白突变引起的疾病的药物中的用途。
5.根据权利要求1或3所述的用途,其特征在于,所述由NLRP3炎症小体引起的疾病为Ⅱ型糖尿病、痛风、阿尔兹海默症、动脉粥样硬化、帕金森综合征、多发性硬化症、肌萎缩侧所硬化症、哮喘、慢性阻塞性肺疾病、肾炎、肠炎、肝炎。
6.根据权利要求2或4所述的用途,其特征在于,所述由NLRP3蛋白突变引起的疾病为冷吡啉相关周期性综合征。
7.根据权利要求2或4所述的用途,其特征在于,所述由NLRP3蛋白突变引起的疾病为家族性寒冷型自身炎症性综合征、穆-韦二氏综合征和慢性婴儿神经系统皮肤关节综合征。
8.根据权利要求3或4所述的用途,其特征在于,所述抑制或阻断FAAH与NLRP3之间相互作用的制剂包括3'-胺甲酰基联苯-3-基环己基氨基甲酸酯、鹰嘴豆牙素A、N-苄基棕榈酰胺、N-(4-氯-3-吡啶基)-4-[(2,2-二氟-1,3-苯并二恶茂-5-基)甲基]-1-哌嗪甲酰胺、4-吗啉基-1,2,5-噻二唑-3-基环辛基(甲基)氨基甲酸酯和N-3-吡啶嗪基-4-[[3-[[5-(三氟甲基)-2-吡啶基]氧基]苯基]亚甲基]-1-哌啶甲酰胺中的至少一种。
9.根据权利要求3或4所述的用途,其特征在于,所述抑制或阻断FAAH与NLRP3之间相互作用的制剂包括3'-胺甲酰基联苯-3-基环己基氨基甲酸酯。
10.根据权利要求3或4所述的用途,其特征在于,所述抑制或阻断FAAH与NLRP3之间相互作用的制剂包括氨基酸序列如SEQ ID NO:1所示的多肽。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110800301.2A CN115616216A (zh) | 2021-07-15 | 2021-07-15 | 抑制或阻断faah与nlrp3之间相互作用的制剂的用途 |
PCT/CN2021/107643 WO2023283977A1 (zh) | 2021-07-15 | 2021-07-21 | 抑制或阻断faah与nlrp3之间相互作用的制剂的用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110800301.2A CN115616216A (zh) | 2021-07-15 | 2021-07-15 | 抑制或阻断faah与nlrp3之间相互作用的制剂的用途 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN115616216A true CN115616216A (zh) | 2023-01-17 |
Family
ID=84855745
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110800301.2A Pending CN115616216A (zh) | 2021-07-15 | 2021-07-15 | 抑制或阻断faah与nlrp3之间相互作用的制剂的用途 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN115616216A (zh) |
WO (1) | WO2023283977A1 (zh) |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AR084433A1 (es) * | 2010-12-22 | 2013-05-15 | Ironwood Pharmaceuticals Inc | Inhibidores de la faah y composiciones farmaceuticas que los contienen |
WO2014004676A1 (en) * | 2012-06-26 | 2014-01-03 | Ironwood Pharmaceuticals, Inc. | Use of faah inhibitors as neuroprotective agents in the cns |
CN103006631B (zh) * | 2012-12-28 | 2015-03-25 | 武汉华士特工业生物技术开发有限公司 | N-苄基脂肪酰胺类化合物在制备神经保护药物中的应用 |
EP3095444A1 (en) * | 2015-05-20 | 2016-11-23 | Dublin City University | A method of treating peripheral inflammatory disease |
TWI743047B (zh) * | 2015-08-17 | 2021-10-21 | 安成生物科技股份有限公司 | 使用雙醋瑞因或其類似物抑制asc表現、nlrp3表現、以及/或nlrp3發炎體複合物的形成之方法 |
WO2019023147A1 (en) * | 2017-07-24 | 2019-01-31 | IFM Tre, Inc. | COMPOUNDS AND COMPOSITIONS FOR THE TREATMENT OF PATHOLOGICAL CONDITIONS ASSOCIATED WITH NLRP ACTIVITY |
GB201712282D0 (en) * | 2017-07-31 | 2017-09-13 | Nodthera Ltd | Selective inhibitors of NLRP3 inflammasome |
GB201905265D0 (en) * | 2019-04-12 | 2019-05-29 | Inflazome Ltd | Inflammasome inhibition |
-
2021
- 2021-07-15 CN CN202110800301.2A patent/CN115616216A/zh active Pending
- 2021-07-21 WO PCT/CN2021/107643 patent/WO2023283977A1/zh unknown
Also Published As
Publication number | Publication date |
---|---|
WO2023283977A1 (zh) | 2023-01-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Gasse et al. | Uric acid is a danger signal activating NALP3 inflammasome in lung injury inflammation and fibrosis | |
Abderrazak et al. | NLRP3 inflammasome: from a danger signal sensor to a regulatory node of oxidative stress and inflammatory diseases | |
Orban et al. | Caffeic acid phenethyl ester induces leukocyte apoptosis, modulates nuclear factor-kappa B and suppresses acute inflammation | |
Zhao et al. | A critical role for interleukin-1β in the progression of autoimmune diseases | |
ES2245028T3 (es) | Uso de lactoferrina en el tratamiento de trastornos inducidos por alergenos. | |
JP6603207B2 (ja) | ヌトリン3aおよびペプチドを用いた肺線維症の阻害 | |
Pastor-Soler et al. | Metformin improves relevant disease parameters in an autosomal dominant polycystic kidney disease mouse model | |
JP7126940B2 (ja) | 補体活性のモジュレーター | |
US20210000927A1 (en) | Modulators of complement activity | |
Sanz et al. | Nimodipine inhibits IL‐1β release stimulated by amyloid β from microglia | |
JP2020500897A (ja) | 補体活性のモジュレータ | |
AU2014249456A1 (en) | Use of levocetirizine and montelukast in the treatment of autoimmune disorders | |
Moshkovits et al. | CMRF35-like molecule 1 (CLM-1) regulates eosinophil homeostasis by suppressing cellular chemotaxis | |
Rose et al. | Transglutaminase 2 limits murine peritoneal acute gout‐like inflammation by regulating macrophage clearance of apoptotic neutrophils | |
Chen et al. | Nrf2 plays a pivotal role in protection against burn trauma-induced intestinal injury and death | |
JP2014516990A (ja) | シータデフェンシンによる炎症性プロテアーゼの遮断 | |
La Manna et al. | Proteomimetics of natural regulators of JAK–STAT pathway: novel therapeutic perspectives | |
Kotas et al. | A role for IL-33–activated ILC2s in eosinophilic vasculitis | |
Qijun et al. | The levels and significance of inflammasomes in the mouse retina following optic nerve crush | |
Girard et al. | Suppressing protein Z‐dependent inhibition of factor Xa improves coagulation in hemophilia A | |
CN115616216A (zh) | 抑制或阻断faah与nlrp3之间相互作用的制剂的用途 | |
US8618113B2 (en) | Treatment for demyelinating disease | |
EP2344149A1 (en) | Use of a compound capable of reducing the uric acid level for the prevention and/or the treatment of lung inflammation and fibrosis | |
JP2009545624A (ja) | 自己免疫またはアレルギー疾患の治療における線形動物由来のシスタチン | |
Tannu et al. | Dual bronchodilatory and pulmonary anti‐inflammatory activity of RO5024118, a novel agonist at vasoactive intestinal peptide VPAC2 receptors |
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 |