CN115969840A - 马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用 - Google Patents
马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用 Download PDFInfo
- Publication number
- CN115969840A CN115969840A CN202211727460.5A CN202211727460A CN115969840A CN 115969840 A CN115969840 A CN 115969840A CN 202211727460 A CN202211727460 A CN 202211727460A CN 115969840 A CN115969840 A CN 115969840A
- Authority
- CN
- China
- Prior art keywords
- pla
- aristolochic acid
- acid iva
- tnf
- iva
- 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
- PADIFGYTAXNCRK-UHFFFAOYSA-N aristolochic acid D Chemical compound C1=C([N+]([O-])=O)C2=C(C(O)=O)C=C3OCOC3=C2C2=C1C(OC)=CC(O)=C2 PADIFGYTAXNCRK-UHFFFAOYSA-N 0.000 title claims abstract description 134
- 239000003814 drug Substances 0.000 title claims abstract description 20
- 208000023275 Autoimmune disease Diseases 0.000 title claims abstract description 17
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims abstract description 8
- 239000003112 inhibitor Substances 0.000 claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 206010039073 rheumatoid arthritis Diseases 0.000 claims abstract description 7
- 230000008685 targeting Effects 0.000 claims abstract description 6
- 229940046728 tumor necrosis factor alpha inhibitor Drugs 0.000 claims abstract description 6
- 239000002452 tumor necrosis factor alpha inhibitor Substances 0.000 claims abstract description 6
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 5
- 229920000747 poly(lactic acid) Polymers 0.000 claims description 58
- 239000004626 polylactic acid Substances 0.000 claims description 58
- 238000011282 treatment Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000003085 diluting agent Substances 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 2
- 239000000839 emulsion Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 239000006188 syrup Substances 0.000 claims description 2
- 235000020357 syrup Nutrition 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 238000009472 formulation Methods 0.000 claims 2
- 239000000314 lubricant Substances 0.000 claims 1
- 238000002474 experimental method Methods 0.000 abstract description 12
- 206010003246 arthritis Diseases 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract description 2
- 150000002611 lead compounds Chemical class 0.000 abstract description 2
- 206010007269 Carcinogenicity Diseases 0.000 abstract 1
- 230000007670 carcinogenicity Effects 0.000 abstract 1
- 231100000260 carcinogenicity Toxicity 0.000 abstract 1
- 230000007674 genetic toxicity Effects 0.000 abstract 1
- 231100000025 genetic toxicology Toxicity 0.000 abstract 1
- 229930014626 natural product Natural products 0.000 abstract 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 17
- 241000699670 Mus sp. Species 0.000 description 15
- 239000002158 endotoxin Substances 0.000 description 15
- 229920006008 lipopolysaccharide Polymers 0.000 description 15
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 12
- 229920003045 dextran sodium sulfate Polymers 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 241000270295 Serpentes Species 0.000 description 5
- 206010009887 colitis Diseases 0.000 description 5
- 230000002757 inflammatory effect Effects 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 description 4
- 108010064785 Phospholipases Proteins 0.000 description 4
- 102000015439 Phospholipases Human genes 0.000 description 4
- -1 but not limited to Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 239000004359 castor oil Substances 0.000 description 3
- 235000019438 castor oil Nutrition 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 3
- 210000004969 inflammatory cell Anatomy 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 206010003694 Atrophy Diseases 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 108090001005 Interleukin-6 Proteins 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 241000078511 Microtome Species 0.000 description 2
- 238000002123 RNA extraction Methods 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- 230000037444 atrophy Effects 0.000 description 2
- BLFLLBZGZJTVJG-UHFFFAOYSA-N benzocaine Chemical compound CCOC(=O)C1=CC=C(N)C=C1 BLFLLBZGZJTVJG-UHFFFAOYSA-N 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000002224 dissection Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000003628 erosive effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 238000003032 molecular docking Methods 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000726094 Aristolochia Species 0.000 description 1
- 241000046617 Aristolochia debilis Species 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 208000012671 Gastrointestinal haemorrhages Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 101000619805 Homo sapiens Peroxiredoxin-5, mitochondrial Proteins 0.000 description 1
- 101000619708 Homo sapiens Peroxiredoxin-6 Proteins 0.000 description 1
- 101000595800 Homo sapiens Phospholipase A and acyltransferase 3 Proteins 0.000 description 1
- 101000987238 Homo sapiens Platelet-activating factor acetylhydrolase 2, cytoplasmic Proteins 0.000 description 1
- 101001072749 Homo sapiens Post-GPI attachment to proteins factor 6 Proteins 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 101150022234 Oc90 gene Proteins 0.000 description 1
- 102100031915 Otoconin-90 Human genes 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 102100022239 Peroxiredoxin-6 Human genes 0.000 description 1
- 102100036066 Phospholipase A and acyltransferase 3 Human genes 0.000 description 1
- 102100027932 Platelet-activating factor acetylhydrolase 2, cytoplasmic Human genes 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102100036591 Post-GPI attachment to proteins factor 6 Human genes 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000008578 acute process Effects 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical group [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 235000011132 calcium sulphate Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 230000027288 circadian rhythm Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000019700 dicalcium phosphate Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 239000005414 inactive ingredient Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 229960000598 infliximab Drugs 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 235000014380 magnesium carbonate Nutrition 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000013227 male C57BL/6J mice Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 125000005395 methacrylic acid group Chemical group 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000031990 negative regulation of inflammatory response Effects 0.000 description 1
- 231100001223 noncarcinogenic Toxicity 0.000 description 1
- 231100000804 nongenotoxic Toxicity 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- 239000003358 phospholipase A2 inhibitor Substances 0.000 description 1
- 150000003904 phospholipids Chemical group 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 102200006537 rs121913529 Human genes 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 208000026775 severe diarrhea Diseases 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000002435 venom Substances 0.000 description 1
- 210000001048 venom Anatomy 0.000 description 1
- 231100000611 venom Toxicity 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 229940100445 wheat starch Drugs 0.000 description 1
Images
Abstract
本发明公开了马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用，属于生物医药领域。其中自身免疫性疾病包括但不限于炎症性肠病和类风湿关节炎，马兜铃酸IVa的分子式为C17H11NO8，分子式为
同时马兜铃酸IVa还用于制备PLA2抑制剂或TNFα抑制剂。本发明通过体外细胞实验和动物实验发现，马兜铃酸IVa可以通过靶向PLA2具有明显的TNFα抑制作用，可作为先导化合物用于制备治疗关节炎和炎性肠病的药物，也可以用于制备PLA2抑制剂或TNFα抑制剂。此外，马兜铃酸IVa是一种天然、无遗传毒性、无致癌性的化合物，在医药制备领域具有潜在的应用价值。
Description
技术领域
本发明属于医药制备领域，特别涉及马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用，特别是马兜铃酸IVa通过靶向结合磷脂酶A2抑制TNFα用于治疗炎症性肠病和类风湿关节炎。
背景技术
磷脂酶A2(PLA2)是一类催化磷脂sn-2位酰基水解的酶超家族，是内源性花生四烯酸、溶血卵磷脂等炎性介质形成的限速酶，广泛参与急、慢性炎症过程。PLA2主要可分为分泌型、胞质型、和非钙离子依赖型等，PLA2抑制剂具有抗炎、抗过敏作用。
自身免疫性疾病是一类由于免疫系统失调引起的慢性疾病，其发病机制尚不明确，主要包括炎症性肠病和类风湿性关节炎等，其主要治疗策略为缓解患者症状，延长缓解期。患者病理上多表现为靶器官(如肠道或关节)炎性细胞大量浸润和炎性细胞因子相关表达增高。肿瘤坏死因子α(TNFα)是一类重要的促炎细胞因子，在临床疾病，尤其是自身免疫性疾病中发挥重要作用，临床上多采用抑制患者体内TNFα表达来缓解患者自身免疫性疾病症状的重要手段。
PLA2抑制剂在治疗动脉粥样硬化、炎性疾病和肾脏疾病中展示出巨大潜力，多项临床试验正在进行中。抗TNFα生物制剂(如英夫利息单抗、高利单抗)是目前自身免疫性疾病的临床一线用药，可通过靶向抑制TNFα达到缓解患者症状的目的。抗TNFα生物制剂价格昂贵，一般通过肌注摄入患者体内。临床研究显示，约有30％-40％患者对抗TNFα生物制剂不敏感，而且抗TNFα生物制剂可能会提高患者感染与结核发病风险。
马兜铃植物具有抗炎镇痛的效果，用药方式有外敷和口服等治疗方式，但其发挥药效的具体机制未知。马兜铃酸IVa(AAIVa)是马兜铃植物中的天然成分，研究表明AAIVa无明显毒性，具有较好的安全性，目前尚未有AAIVa用于治疗自身免疫性疾病如结肠炎或关节炎的相关报道。
发明内容
本发明的目的是提供马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用，特别是马兜铃酸IVa通过靶向结合磷脂酶A2抑制TNFα用于治疗炎症性肠病和类风湿关节炎。
为实现上述目的，本发明采用如下技术方案：
本发明的第一方面提供马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用，所述自身免疫性疾病包括但不限于炎症性肠病和类风湿关节炎，其中马兜铃酸IVa的分子式为C17H11NO8，分子式为
作为优选，所述药物以马兜铃酸IVa作为活性成分，应用剂量为0.1–100uM，更优选为50–100uM。
作为优选，所述药物还包括药学可接受的载体或稀释剂，“药学上可接受的载体”指药物中的非活性成分，包括但不限于碳酸钙、磷酸钙、各种糖例如乳糖、甘露醇等、淀粉、环糊精、硬脂酸镁、纤维素、碳酸镁、丙烯酸聚合物或甲基丙烯酸聚合物、凝胶、水、聚乙二醇、丙二醇、乙二醇、蓖麻油或氢化蓖麻油或多乙氧基氢化蓖麻油、芝麻油、玉米油、花生油。
“药学上可接受的稀释剂”包括但不限于淀粉(如玉米淀粉、小麦淀粉、马铃薯淀粉等)、乳糖、糊精、蔗糖、预胶化淀粉、微晶纤维素、无机盐类(如磷酸氢钙、硫酸钙、残酸钙等)和甘露醇。
作为优选，所述药物选自片剂、胶囊剂、颗粒剂、滴丸剂、混悬剂、糖浆剂、敷剂、肠溶制剂、乳剂悬浮液和注射剂中的至少一种制剂形式。
本发明的第二方面提供马兜铃酸IVa在制备PLA2抑制剂中的应用，应用剂量为0.1–100uM，更优选为50–100uM。
作为优选，PLA2为PLA2R1亚型。
本发明的第三方面提供马兜铃酸IVa在制备TNFα抑制剂中的应用，马兜铃酸IVa通过靶向结合磷脂酶A2抑制TNFα，应用剂量为0.1–100uM，更优选为50–100uM。
与现有技术相比，本发明具有以下有益效果：
(1)本发明通过体外细胞实验和动物实验发现，马兜铃酸IVa可以通过靶向PLA2抑制TNFα，且具有明显的抑制作用，可作为先导化合物用于制备治疗自身免疫性疾病如关节炎和炎性肠病的药物，也可以用于制备PLA2抑制剂或TNFα抑制剂。
(2)本发明中AAIVa是一种天然、无遗传毒性、无致癌性的化合物，本发明显示其可不仅具有开发成为新一代自身免疫性疾病如类风湿性关节炎、炎性肠病的治疗药物的潜力，同时在PLA2或TNFα的靶向抑制剂方面具有潜在的应用价值。
附图说明
图1为实施例中AAs与蛇源PLA2结合能的计算结果。
图2为实施例中AlphaFold 2预测与AAIVa结合的PLA2亚型前三个的结合位点。
图3为实施例中AAIVa靶向结合PLA2R1的实验验证结果。
图4为实施例中AAIVa对LPS刺激后RAW 264.7细胞上清中炎症因子含量的影响。
图5为实施例中AAIVa在TNF-IRES-Luc小鼠体内降低LPS诱导的全身炎症的实验验证结果。
图6为实施例中AAIVa减轻DSS诱导小鼠溃疡性结肠炎的实验验证结果。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚，下面结合本发明实施例的附图，对本发明实施例的技术方案进行清楚、完整地描述。显然，所描述的实施例是本发明的一部分实施例，而不是全部的实施例。基于所描述的本发明的实施例，本领域普通技术人员在无需创造性劳动的前提下所获得的所有其它实施例，都属于本发明保护的范围。
实施例1计算机预测AAIVa抑制蛇源PLA2的能力强于AAI
实验方法：AAI、9-OH AAI、AAIVa与蛇源PLA2(PDB ID:1FV0)结合的能力用GROMACS(CHARMM27 force field)计算，Swiss server(http://www.swissparam.ch)用以制备拓扑兼容。
实验结果：文献报道AAs可以抑制蛇毒PLA2活性，并计算9-OH AAI与蛇源PLA2的结合能与结合位点，如图1所示，蛇源PLA2属于分泌型PLA2，对该蛋白进行类似的计算和预测发现，R1位的羟基(图1黄色箭头)有利于增大结合能，显示AAⅣa比AAI和9-OH AAI具有更高的结合能，提示其抑制PLA2能力更强。
实施例2基于AlphaFold 2预测与AAIVa结合的PLA2亚型鉴定
实验方法：将AlphaFold2预测的蛋白结构数据库中所有的人源PLA2共28种亚型：PLA2G4A、PLA2G10、PLA2G6、PLA2G2A、PLA2G7、PLA2G1B、PLA2G15、PLA2G3、PRDX6、PLA2G5、PLAAT3、PLA2G2C、PLA2G4F、PLA2G4E、PLA2G4B、OC90、PLA2R1、LYPLA2、PLA2G4D、PNPLA2、PAFAH2、PLA2G12B、PLA2G12A、PLA2G2F、PGAP6、PLA2G2E、PLA2G2D、PLA2G4C进行分子对接。
实验结果：将AlphaFold2蛋白结构数据库中所有人类PLA2亚型共计28种，与AAIVa进行分子对接，计算AAIVa与PLA2亚型结合反应能，如图2所示，亲和力最强的前三个分别为PLA2R1(-8.5kcal/mol)、PLA2G6(-8.6kcal/mol)和PLA2G1B(-7.6kcal/mol)。
实施例3AAIVa靶向结合PLA2R1的实验验证
实验试剂：AAIVa(CAS:17413-38-6,Lot:12645)购自上海诗丹德公司。
实验材料：RAW264.7细胞购自上海中国科学院细胞库。
实验仪器：Bio-Rad GelDox XR。
实验方法：DMSO或AAIVa处理RAW 264.7 2h后，收集细胞，用PBS清洗2次后，将细胞重悬到1mL体积。将细胞等体积分为4管细胞后，将DMSO/AAIVa分别用不同梯度温度孵育。温度孵育后化冻速溶三次，收集上清进行Western blotting实验。
实验结果：如图3所示，AAIVa处理后，PLA2R1稳定性增加，结果显示AAIVa靶向PLA2R1。
实施例4AAIVa在RAW264.7中降低TNFα表达
实验试剂：脂多糖(LPS)购自Sigma-Aldrich，Lot号：L2630。
通用型RNA提取裂解液(Lot号：9109)购自Takara。
逆转录试剂盒：HiScript II Q RT SuperMix(Lot号：R223-01)购自诺唯赞。
PCR试剂：ChamQ Universal SYBR qPCR Master Mix(Q711-02)购自诺唯赞。
AAIVa购自上海诗丹德公司。
实验材料：RAW264.7细胞购自上海中国科学院细胞库。
实验仪器：罗氏
480PCR仪器；赛默飞二氧化碳培养箱，HERA Cell150i
实验方法：经LPS诱导后的Raw264.7是一个经典的细胞炎症模型，细胞融合度80％–90％时在六孔板中种板。用AAⅠ与AAIVa处理细胞2h后，吸走上清，撤去AAⅠ与AAIVa。再用LPS(150ng/mL)处理细胞6h后，轻柔吹打下细胞，1000rpm 3min离心后，保留上清用以测定TNFα含量，加入1mL通用型RNA提取裂解液用于qRT PCR。
统计方法：数据均表示为均数±标准误，单因素方差分析用以计算显著性差异，p<0.05被认为有统计学意义。
实验结果：如图4所示，AAIVa降低LPS诱导后RAW 264.7细胞中的TNFα与IL-6含量，与空白对照组相比，LPS处理细胞约升高6倍TNFα含量，而AAIVa预处理则只升高了约4.3倍(AAIVa,50μM)、3倍(AAIVa,100μM)TNFα含量，预处理AAIVa(50,100μM)可显著降低IL-6在LPS诱导后RAW 264.7细胞上清中的含量。
实施例5AAIVa在TNF-IRES-Luc小鼠体内降低TNFα荧光强度
实验试剂：同实施例4。
实验材料：三月龄的SPF级别的TNF-IRES-Luc雄性C57BL/6J小鼠，饲养于SPF级条件，适应性喂养一周后进行实验。
实验仪器：IVIS Lumina Series III system(Perkin Elmer,WLM,USA)。
实验方法：TNF-IRES-Luc小鼠提前45min给0.9％生理盐水，或AAIVa(100mg/kg,i.g.)后，所有小鼠均腹腔注射LPS(n＝3)。在0h、30min、3h、6h、8h、24h观察TNF荧光强度变化。
统计方法：同实施例4。
实验结果：如图5所示，AAIVa在TNF-IRES-Luc小鼠中降低LPS引起的TNFα荧光强度，提示AAIVa抗炎起效快且时间短。所有小鼠在0h本底TNFα荧光强度均在5-9×107(p/s)，证明所有小鼠本底TNFα表达水平相同(图5b)。LPS刺激后，TNFα荧光强度在3h达到峰值(～8.7×108p/s)。AAIVa 100mg/kg在3h显著降低LPS诱导的TNFα荧光强度(～2.9×108p/s)，但AAIVa在6、8、24h并未降低LPS诱导的TNFα荧光强度。同时，阳性对照地塞米松在检测的各个时间点都显著性降低LPS刺激后TNFα荧光强度。
实施例6AAIVa缓解葡聚糖硫酸钠诱导的结肠炎
实验试剂：葡聚糖硫酸钠(DSS,CAS:9011-18-1)购自MP Biomedicals公司，货号S2839。
AAIVa(CAS:17413-38-6,Lot:12645)购自上海诗丹德公司。
实验材料：鼠龄为八周左右的SPF级别C57BL/6J雄性小鼠购自北京维通利华公司，小鼠饲养于温度24-26℃，湿度40％-60％左右，采用12小时明暗昼夜节律照明的清洁级条件，小鼠适应性饲养三天后进行实验。
实验仪器：病理切片机(RM2016)购自徕卡公司，组织摊片机(KD-P)购自科迪仪器。
实验方法：小鼠随机分为两组：DSS造模组，DSS造模+AAIVa治疗组。一组予以3.5％DSS溶液并20mL/kg/天0.9％生理盐水灌胃(n＝5)，一组予以AAIVa 15mg/kg/天20mL/kg灌胃(n＝5)。小鼠每天称重，第十一天后进行解剖并取材。
统计方法：数据均表示为均数±标准误，单因素方差分析用以计算显著性差异，p<0.05被认为有统计学意义。
实验结果：如图6所示，AAIVa可显著缓解DSS引起的小鼠肠炎，提示其可用于治疗结肠炎。仅饮用DSS溶液的小鼠在第7天出现严重腹泻、便血等急性结肠炎现象，解剖可见结肠缩短(图6a,b)，病理可见DSS造模组小肠结构萎缩，局部糜烂并伴有大量炎性细胞浸润(图6e)。与DSS造模组相比，处理AAIVa 15mg/kg的DSS小鼠的便血情况更为轻微、体重下降较少，解剖可见结肠长度增加(图6a,b)。AAIVa+DSS组小鼠病理肠道结构清晰，无糜烂萎缩，仅有少量炎性细胞浸润(图6e)。与DSS造模组相比，AAIVa+DSS组显著降低结肠炎病理评分，AAIVa+DSS组小肠组织中炎症因子TNFα水平显著低于DSS造模组(图6c)。
以上所述为本发明的较佳实施例而已，但本发明不应该局限于该实施例所公开的内容。所以凡是不脱离本发明所公开的精神下完成的等效或修改，都落入本发明保护的范围。
Claims (8)
1.马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用，所述自身免疫性疾病包括但不限于炎症性肠病和类风湿关节炎，其中马兜铃酸IVa的分子式为C17H11NO8，分子式为
2.根据权利要求1所述的应用，其特征在于，所述药物以马兜铃酸IVa作为活性成分，应用剂量为0.1–100uM。
3.根据权利要求2所述的应用，其特征在于，所述药物还包括药学可接受的载体或稀释剂，所述药物选自片剂、胶囊剂、颗粒剂、滴丸剂、混悬剂、糖浆剂、敷剂、肠溶制剂、乳剂悬浮液和注射剂中的至少一种制剂形式。
4.马兜铃酸IVa在制备PLA2抑制剂中的应用。
5.根据权利要求4所述的应用，其特征在于，所述PLA2抑制剂以马兜铃酸IVa作为活性成分，应用剂量为0.1–100uM。
6.根据权利要求4所述的应用，其特征在于，PLA2为PLA2R1亚型。
7.马兜铃酸IVa在制备TNFα抑制剂中的应用。
8.根据权利要求7所述的应用，其特征在于，所述TNFα抑制剂以马兜铃酸IVa作为活性成分，马兜铃酸IVa通过靶向结合PLA2抑制TNFα，应用剂量为0.1–100uM。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211727460.5A CN115969840A (zh) | 2022-12-30 | 2022-12-30 | 马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211727460.5A CN115969840A (zh) | 2022-12-30 | 2022-12-30 | 马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115969840A true CN115969840A (zh) | 2023-04-18 |
Family
ID=85964547
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211727460.5A Pending CN115969840A (zh) | 2022-12-30 | 2022-12-30 | 马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115969840A (zh) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114469930A (zh) * | 2022-04-12 | 2022-05-13 | 中国中医科学院中药研究所 | 马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用 |
-
2022
- 2022-12-30 CN CN202211727460.5A patent/CN115969840A/zh active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114469930A (zh) * | 2022-04-12 | 2022-05-13 | 中国中医科学院中药研究所 | 马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用 |
Non-Patent Citations (1)
| Title |
|---|
| WEIYING LIU等: "Non‑carcinogenic/non‑nephrotoxic aristolochic acid IVa exhibited anti‑inflammatory activities in mice", JOURNAL OF NATURAL MEDICINES, vol. 77, no. 2, 16 December 2022 (2022-12-16), pages 251 - 261 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Qi et al. | Hesperidin inhibits synovial cell inflammation and macrophage polarization through suppression of the PI3K/AKT pathway in complete Freund's adjuvant-induced arthritis in mice | |
| US6277836B1 (en) | Methods and compositions for treating inflammatory bowel disease | |
| JP5964821B2 (ja) | 痛風と高尿酸血症の処置 | |
| Fellner et al. | Successful use of cyclophosphamide and prednisone for initial treatment of pemphigus vulgaris | |
| KR20020059416A (ko) | 프로게스테론 또는 이의 유사체들을 유효성분으로함유하는 염증 치료용 약학적 조성물 | |
| Li et al. | Regulatory mechanism of mesalazine on TLR4/MyD88-dependent pathway in mouse ulcerative colitis model. | |
| Tang et al. | The novel hepatoprotective mechanisms of silibinin-phospholipid complex against d-GalN/LPS-induced acute liver injury | |
| CN115969840A (zh) | 马兜铃酸IVa在制备治疗自身免疫性疾病的药物中的应用 | |
| CN112451537A (zh) | 黄芩苷在制备预防和/或治疗无症状高尿酸血症和/或尿酸性肾病的药物中的应用 | |
| Hüseyin et al. | Cyclosporin treatment of anal and perianal lesions associated with Crohn's disease | |
| JPS61106521A (ja) | 血管増殖抑制剤 | |
| US11925634B2 (en) | Use of koumine in preparation of medicament for treatment of inflammatory bowel disease | |
| TWI435727B (zh) | 調節細胞激素分泌之用途 | |
| CN111467327B (zh) | 姜烯酮a在制备防治结肠炎药物方面的应用 | |
| CN108853085A (zh) | 儿茶素类和表儿茶素类化合物用于上调MicroRNA-126表达水平的用途 | |
| CN111281866B (zh) | 一种预防或治疗溃疡性结肠炎的中药化学制剂 | |
| AU2017287461B2 (en) | Application of methyl 4-[9-(6-aminopuryl)]-2(S)-hydroxybutyrate in preparation of medicament for treating psoriasis and vitiligo | |
| CN101336916A (zh) | 茶多酚的用途 | |
| CN105395584B (zh) | 片仔癀及其制剂在制备治疗多发性硬化症的药物中的应用 | |
| CN113304163B (zh) | 三叶豆紫檀苷在治疗关节炎中的用途 | |
| CN116850180B (zh) | 软木花椒素在制备治疗类风湿关节炎的药物中的应用 | |
| CN111184715B (zh) | 佛手苷内酯在制备hiv-1潜伏感染逆转剂中的应用 | |
| CN111067892B (zh) | 青蒿琥酯在治疗原发性硬化性胆管炎模型小鼠的应用 | |
| WO2023110855A1 (en) | Compositions for treating autoimmune arthritis | |
| CN117562910A (zh) | 一种甘氨猪去氧胆酸在制备治疗炎症性肠病的药物中的应用 |
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 |