CN102499917B - 吲哚酮类化合物在制备神经保护药物中的应用 - Google Patents
吲哚酮类化合物在制备神经保护药物中的应用 Download PDFInfo
- Publication number
- CN102499917B CN102499917B CN201110327436.8A CN201110327436A CN102499917B CN 102499917 B CN102499917 B CN 102499917B CN 201110327436 A CN201110327436 A CN 201110327436A CN 102499917 B CN102499917 B CN 102499917B
- Authority
- CN
- China
- Prior art keywords
- μms
- mpp
- nnos
- caused
- mptp
- 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.)
- Expired - Fee Related
Links
- 239000003814 drug Substances 0.000 title claims abstract description 20
- 210000005036 nerve Anatomy 0.000 title claims abstract description 15
- 150000005624 indolones Chemical class 0.000 title abstract 5
- 210000002569 neuron Anatomy 0.000 claims abstract description 21
- 208000018737 Parkinson disease Diseases 0.000 claims abstract description 11
- 150000003839 salts Chemical class 0.000 claims abstract description 10
- 208000015122 neurodegenerative disease Diseases 0.000 claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 7
- 230000000694 effects Effects 0.000 claims description 27
- 102000008299 Nitric Oxide Synthase Human genes 0.000 claims description 15
- 108010021487 Nitric Oxide Synthase Proteins 0.000 claims description 15
- -1 indole ketone compound Chemical class 0.000 claims description 15
- SIKJAQJRHWYJAI-UHFFFAOYSA-N benzopyrrole Natural products C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 claims description 12
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 claims description 12
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 claims description 12
- 230000002401 inhibitory effect Effects 0.000 claims description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 239000002253 acid Substances 0.000 claims description 6
- 230000003287 optical effect Effects 0.000 claims description 6
- 239000008187 granular material Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 230000004770 neurodegeneration Effects 0.000 abstract description 10
- 206010029350 Neurotoxicity Diseases 0.000 abstract description 6
- 206010044221 Toxic encephalopathy Diseases 0.000 abstract description 6
- 125000000649 benzylidene group Chemical group [H]C(=[*])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 abstract description 6
- 230000007135 neurotoxicity Effects 0.000 abstract description 6
- 231100000228 neurotoxicity Toxicity 0.000 abstract description 6
- ROSDSFDQCJNGOL-UHFFFAOYSA-N protonated dimethyl amine Natural products CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 abstract description 6
- 239000000203 mixture Substances 0.000 abstract description 5
- 230000006907 apoptotic process Effects 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 3
- 208000001572 Mycoplasma Pneumonia Diseases 0.000 abstract 2
- 201000008235 Mycoplasma pneumoniae pneumonia Diseases 0.000 abstract 2
- FIRXFHJQGIIJDB-UHFFFAOYSA-N 1-methyl-2,3-dihydroindole Chemical compound C1=CC=C2N(C)CCC2=C1 FIRXFHJQGIIJDB-UHFFFAOYSA-N 0.000 abstract 1
- VSWICNJIUPRZIK-UHFFFAOYSA-N 2-piperideine Chemical compound C1CNC=CC1 VSWICNJIUPRZIK-UHFFFAOYSA-N 0.000 abstract 1
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 46
- 241000252212 Danio rerio Species 0.000 description 43
- 101710111444 Nitric oxide synthase, brain Proteins 0.000 description 29
- 102100022397 Nitric oxide synthase, brain Human genes 0.000 description 28
- 238000000034 method Methods 0.000 description 24
- PLRACCBDVIHHLZ-UHFFFAOYSA-N 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine Chemical compound C1N(C)CCC(C=2C=CC=CC=2)=C1 PLRACCBDVIHHLZ-UHFFFAOYSA-N 0.000 description 23
- 229960003638 dopamine Drugs 0.000 description 23
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 22
- 101001135571 Mus musculus Tyrosine-protein phosphatase non-receptor type 2 Proteins 0.000 description 21
- 230000008569 process Effects 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 18
- 238000012360 testing method Methods 0.000 description 17
- LLDWLPRYLVPDTG-UHFFFAOYSA-N vatalanib succinate Chemical compound OC(=O)CCC(O)=O.C1=CC(Cl)=CC=C1NC(C1=CC=CC=C11)=NN=C1CC1=CC=NC=C1 LLDWLPRYLVPDTG-UHFFFAOYSA-N 0.000 description 14
- 210000004556 brain Anatomy 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 11
- 230000014509 gene expression Effects 0.000 description 11
- JYGFTBXVXVMTGB-UHFFFAOYSA-N indolin-2-one Chemical compound C1=CC=C2NC(=O)CC2=C1 JYGFTBXVXVMTGB-UHFFFAOYSA-N 0.000 description 10
- 241000700159 Rattus Species 0.000 description 9
- 230000033001 locomotion Effects 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 8
- 230000019581 neuron apoptotic process Effects 0.000 description 8
- 101000944124 Homo sapiens Cingulin Proteins 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 101710090055 Nitric oxide synthase, endothelial Proteins 0.000 description 6
- 102100028452 Nitric oxide synthase, endothelial Human genes 0.000 description 6
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 6
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 6
- 230000006399 behavior Effects 0.000 description 6
- 230000002490 cerebral effect Effects 0.000 description 6
- 230000006378 damage Effects 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 230000000324 neuroprotective effect Effects 0.000 description 6
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical group OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 5
- 101150076211 TH gene Proteins 0.000 description 5
- 230000008499 blood brain barrier function Effects 0.000 description 5
- 210000001218 blood-brain barrier Anatomy 0.000 description 5
- 230000007547 defect Effects 0.000 description 5
- 210000001161 mammalian embryo Anatomy 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 230000002441 reversible effect Effects 0.000 description 5
- 230000009182 swimming Effects 0.000 description 5
- DYLIWHYUXAJDOJ-OWOJBTEDSA-N (e)-4-(6-aminopurin-9-yl)but-2-en-1-ol Chemical compound NC1=NC=NC2=C1N=CN2C\C=C\CO DYLIWHYUXAJDOJ-OWOJBTEDSA-N 0.000 description 4
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 4
- 208000027089 Parkinsonian disease Diseases 0.000 description 4
- 206010034010 Parkinsonism Diseases 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 238000007689 inspection Methods 0.000 description 4
- 229960004502 levodopa Drugs 0.000 description 4
- 230000001537 neural effect Effects 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 208000012661 Dyskinesia Diseases 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 102000010909 Monoamine Oxidase Human genes 0.000 description 3
- 108010062431 Monoamine oxidase Proteins 0.000 description 3
- 108091000117 Tyrosine 3-Monooxygenase Proteins 0.000 description 3
- 102000048218 Tyrosine 3-monooxygenases Human genes 0.000 description 3
- 238000000540 analysis of variance Methods 0.000 description 3
- 230000003527 anti-angiogenesis Effects 0.000 description 3
- 210000004204 blood vessel Anatomy 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000007928 intraperitoneal injection Substances 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 230000029052 metamorphosis Effects 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- MEZLKOACVSPNER-GFCCVEGCSA-N selegiline Chemical compound C#CCN(C)[C@H](C)CC1=CC=CC=C1 MEZLKOACVSPNER-GFCCVEGCSA-N 0.000 description 3
- 239000002002 slurry Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 210000000278 spinal cord Anatomy 0.000 description 3
- 238000012453 sprague-dawley rat model Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- 239000004475 Arginine Substances 0.000 description 2
- 206010003594 Ataxia telangiectasia Diseases 0.000 description 2
- 101100227322 Caenorhabditis elegans fli-1 gene Proteins 0.000 description 2
- 206010008096 Cerebral atrophy Diseases 0.000 description 2
- 108010077544 Chromatin Proteins 0.000 description 2
- 208000023105 Huntington disease Diseases 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 101100281205 Mus musculus Fli1 gene Proteins 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 208000032319 Primary lateral sclerosis Diseases 0.000 description 2
- 206010037211 Psychomotor hyperactivity Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 206010044565 Tremor Diseases 0.000 description 2
- 206010046298 Upper motor neurone lesion Diseases 0.000 description 2
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 2
- 229960004373 acetylcholine Drugs 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 235000009697 arginine Nutrition 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 210000003483 chromatin Anatomy 0.000 description 2
- 210000003792 cranial nerve Anatomy 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000012744 immunostaining Methods 0.000 description 2
- 201000010901 lateral sclerosis Diseases 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 230000004660 morphological change Effects 0.000 description 2
- 208000005264 motor neuron disease Diseases 0.000 description 2
- 201000006417 multiple sclerosis Diseases 0.000 description 2
- 239000002858 neurotransmitter agent Substances 0.000 description 2
- 229960003753 nitric oxide Drugs 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 208000002320 spinal muscular atrophy Diseases 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 210000003523 substantia nigra Anatomy 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000009261 transgenic effect Effects 0.000 description 2
- 229950000578 vatalanib Drugs 0.000 description 2
- YCOYDOIWSSHVCK-UHFFFAOYSA-N vatalanib Chemical compound C1=CC(Cl)=CC=C1NC(C1=CC=CC=C11)=NN=C1CC1=CC=NC=C1 YCOYDOIWSSHVCK-UHFFFAOYSA-N 0.000 description 2
- 230000007998 vessel formation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- UAKWLVYMKBWHMX-RVDMUPIBSA-N (3e)-3-[[4-(dimethylamino)phenyl]methylidene]-1h-indol-2-one Chemical compound C1=CC(N(C)C)=CC=C1\C=C\1C2=CC=CC=C2NC/1=O UAKWLVYMKBWHMX-RVDMUPIBSA-N 0.000 description 1
- BJHCYTJNPVGSBZ-YXSASFKJSA-N 1-[4-[6-amino-5-[(Z)-methoxyiminomethyl]pyrimidin-4-yl]oxy-2-chlorophenyl]-3-ethylurea Chemical compound CCNC(=O)Nc1ccc(Oc2ncnc(N)c2\C=N/OC)cc1Cl BJHCYTJNPVGSBZ-YXSASFKJSA-N 0.000 description 1
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- CVOFKRWYWCSDMA-UHFFFAOYSA-N 2-chloro-n-(2,6-diethylphenyl)-n-(methoxymethyl)acetamide;2,6-dinitro-n,n-dipropyl-4-(trifluoromethyl)aniline Chemical compound CCC1=CC=CC(CC)=C1N(COC)C(=O)CCl.CCCN(CCC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C=C1[N+]([O-])=O CVOFKRWYWCSDMA-UHFFFAOYSA-N 0.000 description 1
- DBERHVIZRVGDFO-UHFFFAOYSA-N Acetoxyacetone Chemical compound CC(=O)COC(C)=O DBERHVIZRVGDFO-UHFFFAOYSA-N 0.000 description 1
- 238000012935 Averaging Methods 0.000 description 1
- 102000000584 Calmodulin Human genes 0.000 description 1
- 108010041952 Calmodulin Proteins 0.000 description 1
- 101100269062 Danio rerio actba gene Proteins 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 101000974009 Homo sapiens Nitric oxide synthase, brain Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 0.000 description 1
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000008109 Mixed Function Oxygenases Human genes 0.000 description 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 description 1
- 229940086616 Monoamine oxidase B inhibitor Drugs 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 108010083674 Myelin Proteins Proteins 0.000 description 1
- 102000006386 Myelin Proteins Human genes 0.000 description 1
- VLQGDKKHHCKIOJ-UHFFFAOYSA-N NNOS Chemical compound NNOS VLQGDKKHHCKIOJ-UHFFFAOYSA-N 0.000 description 1
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 1
- 206010056677 Nerve degeneration Diseases 0.000 description 1
- 208000028389 Nerve injury Diseases 0.000 description 1
- 206010029240 Neuritis Diseases 0.000 description 1
- 102000006538 Nitric Oxide Synthase Type I Human genes 0.000 description 1
- 108010008858 Nitric Oxide Synthase Type I Proteins 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000004659 Presynaptic Receptors Human genes 0.000 description 1
- 108010003717 Presynaptic Receptors Proteins 0.000 description 1
- 238000011530 RNeasy Mini Kit Methods 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 229940091171 VEGFR-2 tyrosine kinase inhibitor Drugs 0.000 description 1
- 102000016549 Vascular Endothelial Growth Factor Receptor-2 Human genes 0.000 description 1
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- BEVHTVRRVVEMEF-UHFFFAOYSA-N [6'-acetyloxy-4-amino-2',7'-difluoro-5-(methylamino)-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl] acetate Chemical compound C12=CC(F)=C(OC(C)=O)C=C2OC2=CC(OC(C)=O)=C(F)C=C2C21OC(=O)C1=C(N)C(NC)=CC=C21 BEVHTVRRVVEMEF-UHFFFAOYSA-N 0.000 description 1
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001130 astrocyte Anatomy 0.000 description 1
- 210000004227 basal ganglia Anatomy 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 230000031709 bromination Effects 0.000 description 1
- 238000005893 bromination reaction Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000003986 cell retinal photoreceptor Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000001713 cholinergic effect Effects 0.000 description 1
- 210000002932 cholinergic neuron Anatomy 0.000 description 1
- 235000013477 citrulline Nutrition 0.000 description 1
- 229960002173 citrulline Drugs 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000027326 copulation Effects 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000006114 decarboxylation reaction Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 210000002451 diencephalon Anatomy 0.000 description 1
- 229940052764 dopaminergic anti-parkinson drug mao b inhibitors Drugs 0.000 description 1
- 210000005064 dopaminergic neuron Anatomy 0.000 description 1
- 230000003291 dopaminomimetic effect Effects 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 210000003278 egg shell Anatomy 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- FVTCRASFADXXNN-SCRDCRAPSA-N flavin mononucleotide Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-SCRDCRAPSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000000574 ganglionic effect Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 235000003969 glutathione Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 102000055707 human NOS1 Human genes 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000001259 mesencephalon Anatomy 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 230000008811 mitochondrial respiratory chain Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000002969 morbid Effects 0.000 description 1
- 210000005012 myelin Anatomy 0.000 description 1
- 230000018791 negative regulation of catalytic activity Effects 0.000 description 1
- 210000001577 neostriatum Anatomy 0.000 description 1
- 230000008764 nerve damage Effects 0.000 description 1
- 230000002887 neurotoxic effect Effects 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000001898 pallidal effect Effects 0.000 description 1
- 208000021090 palsy Diseases 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000003518 presynaptic effect Effects 0.000 description 1
- 208000037920 primary disease Diseases 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- FNKQXYHWGSIFBK-RPDRRWSUSA-N sapropterin Chemical compound N1=C(N)NC(=O)C2=C1NC[C@H]([C@@H](O)[C@@H](O)C)N2 FNKQXYHWGSIFBK-RPDRRWSUSA-N 0.000 description 1
- 229960004617 sapropterin Drugs 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 229960003946 selegiline Drugs 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000000862 serotonergic effect Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 208000027765 speech disease Diseases 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 238000011477 surgical intervention Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000033912 thigmotaxis Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000005760 tumorsuppression Effects 0.000 description 1
- 229940121358 tyrosine kinase inhibitor Drugs 0.000 description 1
- 239000005483 tyrosine kinase inhibitor Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- A61K31/404—Indoles, e.g. pindolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/54—Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
- A61K35/545—Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular 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
-
- 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
-
- 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
Abstract
本发明公开了吲哚酮类化合物在制备治疗神经保护药物中的应用，其中所述的吲哚酮类化合物选自以下化合物或药学上可接受的盐或它们的组合物：3-[4-(二甲胺)苯亚甲基]二氢吲哚-2-酮和3-(3，′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2。本发明研究结果表明，以上两种吲哚酮类化合物能够有效阻止由MPP+和MPTP所致的神经毒性，防止神经细胞的凋亡，具有突出的神经保护作用，尤其是对中枢神经的保护作用，可应用于预防或治疗包括帕金森病的多种神经退行性疾病。
Description
技术领域
本发明属于生化医药领域，具体涉及吲哚酮类化合物在制备神经保护药物中的应用。
背景技术
神经退行性疾病是一种大脑和脊髓的细胞神经元丧失的疾病状态。大脑和脊髓由神经元组成，神经元有不同的功能，如控制运动、处理感觉信息、并作出决策。通常，大脑和脊髓的细胞是不会再生的，所以过度的损害可能是毁灭性的和不可逆转的。神经退行性疾病是由神经元或其髓鞘的丧失所致，随着时间的推移而恶化，以导致功能障碍。神经退行性疾病包括阿兹海默病、肌肉萎缩性侧索硬化症、共济失调毛细血管扩张症、牛海绵状脑病、克雅二氏病、亨廷顿氏病、小脑萎缩症、多发性硬化症、帕金森病、原发性侧索硬化、脊髓性肌萎缩症等。
帕金森病(Parkinson′s disease，PD)又称“震颤麻痹”、巴金森氏症或柏金逊症，是老年人中常见的神经变性疾病，多在60岁以后发病。主要表现为患者动作缓慢，手脚或身体其它部分的震颤，身体失去柔软性，变得僵硬。在≥65岁人群中，1％患有此病；在＞40岁人群中则为0.4％.本病也可在儿童期或***发病。50％～80％的病例起病隐袭，首发症状通常是一侧手部的4～8Hz的静止性“捻丸样”震颤。言语障碍是帕金森病患者常见症状。
该病主要是因位于中脑部位黑质中的细胞发生病理性改变后，多巴胺的合成减少，抑制乙酰胆碱的功能降低，则乙酰胆碱的兴奋作用相对增强。两者失衡的结果便出现了“震颤麻痹”。PD与纹状体内的多巴胺(DA)含量显著减少有关。目前较公认的学说为“多巴胺学说”和“氧化应激说”。前者指出DA合成减少使纹状体DA含量降低，黑质-纹状体通路多巴胺能与胆碱能神经功能平衡失调，胆碱能神经元活性相对增高，使锥体外系功能亢进，发生震颤性麻痹。后者解释了黑质多巴胺能神经元变性的原因，即在氧化应激时，PD患者DA氧化代谢过程中产生大量H2O2和超氧阴离子，在黑质部位Fe2+催化下，进一步生成毒性更大的羟自由基，而此时黑质线粒体呼吸链的复合物I活性下降，抗氧化物(特别是谷胱甘肽)消失，无法清除自由基，因此，自由基通过氧化神经膜类脂、破坏DA神经元膜功能或直接破坏细胞DNA，最终导所致神经元变性。
目前治疗帕金森病的方法主要通过药物治疗或外科手术。其中药物治疗中最常用也是最有效的药物是左旋多巴制剂。左旋多巴是多巴胺的代谢前体，可以通过血脑屏障，进入基底节后经脱羧而成多巴胺，起着补充多巴胺神经递质缺乏的作用。使用选择性单胺氧化酶-B抑制剂(如司来吉兰)，能够抑制多巴胺的再摄取及突触前受体，增加及延长左旋多巴的效果，但是也难以治愈帕金森病，而且具有不良副作用。应用左旋多巴治疗时，随着治疗时间的延长，会出现药效的波动性(开-关效应)，不良反应包括不自主动作(动作困难)，大体表现为口-面或肢体的舞蹈动作或肌张力障碍。外科治疗主要有神经核团细胞毁损手术和电刺激手术两种方式，其原理是通过抑制脑细胞的异常活动来达到改善症状的目的。通过立体定向切除苍白球的后腹侧部(苍白球切开术)可显著改善“关”状态下的动作过缓以及左旋多巴诱发的动作困难。但是，通过脑神经损毁手术将部分脑神经损毁来控制帕金森病，脑神经一旦损毁就无法复原，而且术后恢复也很困难，只是能控制而不能治愈帕金森病。
1-甲基l-4-苯基吡啶离子(1-methyl-4-pehnylpyridine ion，MPP+)和1-甲基-4-苯基-1，2，3，6-四氢吡啶(1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine，MPTP)具有神经毒性作用，能够诱导PD的产生。MPTP进入体内通过血脑屏障在单胺氧化酶-B(monoamine oxidase B，MAO-B)的作用下转化为相应的1-甲基-4-苯基-2，3-二氢吡啶离子(MPP+)，到细胞外间隙，MPP+通过突触前膜的多巴胺转运体选择性进入黑质神经元内。MPP+在神经元内可抑制酪氨酸羟化酶(tyrosine hydroxylase，TH)的活性从而减少多巴胺的合成，多巴胺是一种抑制性神经递质，如果其抑制作用过分降低，临床上就会出现PD症状。研究表明，MPP+在神经元内还可通过促进一氧化氮合酶释放一氧化氮促进MPTP/MPP+的神经毒性。
3-[4-(二甲胺)苯亚甲基]二氢吲哚-2-酮(SU4312)和3-(3，′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2(SU5416)是在20世纪90年代研发出来的化合物，已知具有抗血管生成作用，可通过阻断向癌细胞的供血来抑制肿瘤生长，现主要应用于抗肿瘤药物中。
经检索国内外文献及专利，未见有关SU4312和SU5416在神经保护方面的报道。
发明内容
本发明的目的是针对以上要解决的技术问题，提供能够起到有效的神经保护的药物。
在本发明的第一方面，提供了吲哚酮类化合物在制备神经保护药物中的应用。
其中，所述的吲哚酮类化合物选自以下化合物、它们的旋光异构体以及它们与药学上可接受酸的加成盐：
3-[4-(二甲胺)苯亚甲基]二氢吲哚-2-酮、3-(3，′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2。
在另一优选例中，以上所述吲哚酮类化合物应用于制备中枢神经保护药物。该神经保护药物优选为适用于治疗各种神经退行性疾病包括但不限于阿兹海默病、肌肉萎缩性侧索硬化症、共济失调毛细血管扩张症、牛海绵状脑病、克雅二氏病、亨廷顿氏病、小脑萎缩症、多发性硬化症、帕金森病、原发性侧索硬化、脊髓性肌萎缩症等。
在另一优选例中，所述神经保护药物用于治疗帕金森病。
在另一优选例中，所述3-[4-(二甲胺)苯亚甲基]二氢吲哚-2-酮、其旋光异构体或药学上可接受酸的加成盐的浓度为0.3-30μM。
进一步，所述3-(3，′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2、其旋光异构体或药学上可接受酸的加成盐的浓度为0.3-30μM。
所述吲哚酮类化合物的酸加成盐优选为药学上可接受的、与适当的酸(例如盐酸、醋酸、硫酸)形成无毒的盐，除了药物上可接受的盐以外，其它的盐也包括在本发明之中。
上述两种吲哚酮类化合物的结构式如下式所示：
(1)3-[4-(二甲胺)苯亚甲基]二氢吲哚-2-酮(SU4312，CAS No：5812-07-7)
(2)3-(3，′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2(SU5416，CAS No.204005-46-9)
上述各化合物均可通过商业途径购买得到，也可按照现有的文献所公开的方法制备得到。
在神经保护药物形式中，除了上述两种吲哚酮类化合物之外，还可存在能防止多巴胺(DA)能神经元损耗进程的其它活性物质。
通常，将本发明的化合物以药物组合物的形式给药，该药物组合物包含上述两种化合物中的至少一种和可药用的赋形剂或稀释剂，这种组合物通常采用与所希望的给药方式相适应的固体或液体赋形剂或稀释剂按常规方法配制。对于口服给药，可制成片剂、颗粒剂、悬浮剂、粉剂、胶囊等形式；对于非经胃肠给药，可以制成注射液、注射悬浮液等形式。
现代科学研究发现，有三种同型的NOS(Nitric Oxide Synthase，一氧化氮合酶)，分别为nNOS、iNOS和eNOS。其中，nNOS是中枢神经***神经元中的主要形式，eNOS主要存在于大脑血管表皮细胞，而iNOS则在星形胶质细胞和小神经细胞中表达。根据本发明的研究结果，3-[4-(二甲胺)苯亚甲基]二氢吲哚-2-酮(SU4312)和3-(3，′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2(SU5416)可通过抑制nNOS来阻止MPP+所致神经毒性。使用试管NOS活性检测，发现SU4312和SU5416能够直接抑制纯化nNOS的酶活性，而且前者对nNOS具有高度选择性，通过非竞争的方式直接抑制nNOS。SU5416除直接抑制NOS活性外，还能抑制nNOS蛋白的表达。
nNOS过度活化导致产生过量NO，也是导致神经元丢失的神经毒性因素。在神经退化紊乱过程中，nNOS常由于兴奋毒性而过度活化。进一步，nNOS抑制剂保护MPTP所致病理；且缺少nNOS基因的突变小鼠比野生型小鼠对MPTP处理更有抵抗力。因此，选择性nNOS抑制剂在治疗PD中具有医疗用途。在另一方面，NO作为一种神经调节物质，也介导许多其他重要的生理过程。例如学习和记忆。具有中度抑制作用的nNOS抑制剂可能具有更高的治疗重要性，因为他们对大脑的生理功能产生很少或不产生干涉。事实上，本发明的实验数据表明SU4312和SU5416可中度抑制nNOS。我们还发现SU4312能够渗透血脑屏障并到达大鼠脑。因此，可将SU4312和SU5416作为治疗神经退化紊乱的药物，其通过对有害的血管生成和NO-介导对神经毒性进行共抑制，从而达到神经保护作用。
附图说明
图1a表示SU4312对MPP+所致神经元凋亡的作用：(A)SU4312和PTK787/ZK222584对MPP+所致神经元凋亡的作用的对比；(B)SU4312对神经元凋亡的阻断作用及对MPP+所致形态变化的逆转作用；(C)SU4312对MPP+所致的细胞核的固缩的逆转作用。
图1b表示SU5416对MPP+所致神经元凋亡的作用：(A)SU5416和PTK787/ZK222584对MPP+所致神经元凋亡的作用的对比；(B)SU5416对神经元凋亡的阻断作用及对MPP+所致形态变化的逆转作用；(C)SU5416对MPP+所致的细胞核的固缩的逆转作用。
图2a表示SU4312对斑马鱼中MPTP所致神经毒性的抑制作用：(A)不同处理组中斑马鱼脑DA神经元的对比；(B)每个处理组的TH+神经元的统计分析结果对比；(C)SU4312逆转MPTP对th基因表达的抑制。
图2b表示SU5416对斑马鱼中MPTP所致神经毒性的抑制作用：(A)不同处理组中斑马鱼脑DA神经元的对比；(B)每个处理组的TH+神经元的统计分析结果对比；(C)SU5416对MPTP所致斑马鱼幼体运动行为缺陷的减弱作用。
图3表示：SU4312使MPTP所致斑马鱼幼体运动行为缺陷减弱：(A)不同处理组的斑马鱼移动轨迹对比；(B)不同处理组中每个斑马鱼幼体的移动总距离的统计分析结果。
图4是斑马鱼中SU5416，SU4312和PTK787/ZK222584之间的抗血管生成作用的比较。
图5表示SU4312，SU5416剂量依赖地抑制MPP+诱导的神经细胞内NO生成。
图6表示SU4312和SU5416在体外抑制nNOS酶的活性：(A)10μM、20μM SU4312与不同浓度L-精氨酸竞争抑制nNOS的反应模式，表现为非竞争性抑制；(B)SU4312抑制nNOS酶活性的Ki常数；(C)SU5416剂量依赖地抑制nNOS酶活性。
图7是SU5416抑制MPP+诱导的神经元中nNOS蛋白表达。
图8是下列物质的HPLC色谱图：(A)标准SU4312溶液(150μg/ml)；(B)来自对照组大鼠的脑匀浆；(C-E)腹腔内注射SU4312(12mg/kg)15min(C)、30min(D)、1h(E)后的大鼠脑匀浆。峰1：顺式SU4312；峰2：反式SU4312。
具体实施方式
下面结合附图和实施例对本发明作进一步的详述，但并不局限于所述具体实施例。本领域普通技术人员在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换，但这些修改和替换均落入本发明的保护范围内。
本发明使用大鼠和斑马鱼作为帕金森病动物模型进行吲哚酮类化合物的治疗作用试验。
通过用Dunnett检验与对照组进行多重比较，或者用Tukey-Kramer分离测试法在多个处理组之间进行比较来分析方差，以确定其统计显著性。可允许的显著性差异为p＜0.05。
实施例1原代小脑颗粒神经元(CGN)的培养
准备8日龄Sprague-Dawley大鼠(香港理工大学动物养殖研究所)，以2.7×105个细胞/毫升的密度将神经元置于改良的伊格尔基础培养基(含有10％胎牛血清、25mM KCl、2mM谷氨酸盐)中培养。
实施例2斑马鱼的培养
按照《斑马鱼手册》(Westerfield，M.(1993)The Zebrafish book ：a guide for the laboratoryuse of zebrafish(Brachydanio rerio)，University of Oregon Press，Eugene.Or.)培养野生型斑马鱼(AB系)和Tg(fli-1：EGFP)转基因斑马鱼。通过自然成对组合交配(3-12月龄)产生斑马鱼胚胎，28.5℃胚胎培养基培养(13.7mM NaCl，540μM KCl，pH 7.4，25μM Na2HDO4，44μMKH2PO4，300μM CaCl2，100μM MgSO4，420μM NaHCO3，pH 7.4)。受精后1日(1dpf)选出健康的斑马鱼胚胎并进行人工剥除蛋壳，分别放进12孔板，每孔20条。
实施例3 SU4312对MPP+所致神经毒性的离体实验
CGN离体培养8日后，用1μM、3μM、10μM、20μM、30μM的SU4312或3μM、10μM、20μM、30μM的SU5416对CGNs进行预处理2h，然后再用35μM MPP+处理24h。对照组不使用任何试剂进行处理。用MTT(3-(4，5-二甲基-2-噻唑)-2，5-二苯基溴化四氮唑噻唑蓝)检测法测定线粒体脱氢酶的活性来估算存活神经元的数量。具体检测过程依照罗氏应用科学MTT试剂盒I说明书的指导进行操作。在96孔平板上培养神经元，每孔(含100μl培养基)加10μl浓度为5mg/ml的MTT标记反应物，将平板在37℃孵化4小时。孵化完成后，每孔中加入100μl溶解液(0.01N HCl用10％SDS溶液配)，放置16-20小时。波长570nm测量吸光度(对照波长为655nm)。
实验结果表明，随着浓度的增加，SU4312和SU5416阻止MPP+所致神经细胞死亡的能力逐渐增强(图1a(A)，图1b(A))。然而，SU4312和SU5416本身并不影响细胞增殖，也不会在实验浓度的反应过程中表现出细胞毒性效应。比较例中使用另一种特异性的VEGFR-2抑制剂PTK787/ZK222584，瓦他拉尼，一种小分子酪氨酸激酶抑制剂)进行测试。由图1a(A)可见，MTT法测定结果表明，PTK787/ZK222584(3-10μM)并不能够阻断神经元的凋亡。
使用20μM SU4312或SU5416对CGNs进行预处理2h，然后接触35μM MPP+。24h后用FDA/PI双染色检测CGN存活情况(10μg/ml FDA和5μg/ml PI孵化15min后，用紫外光显微镜和相差显微镜进行检测和照相，并将两者所得的照片进行比较。)和用Heochst33342对细胞核染色来检测染色质固缩(用冰冷的PBS混合4％甲醛洗脱直径35mm平皿中生长的CGN(2.7×106个细胞)，再用Heochst33342(5μg/ml)于4℃染色5min，荧光显微镜放大400倍)。显微镜下对染色质固缩的死亡细胞核进行计数，结果以占计得总细胞核数的百分比表示。数据取三个相互独立的试验的平均值±方差。显著性差异：(A)中与MPP+实验组比较或(C)中与正常对照组比较*p＜0.05及**p＜0.01；(C)中与MPP+组##p＜0.01(Turkey’s检验)。
相差显微镜和FDA/PI双染色检测结果表明，SU4312和SU5416明显地阻断了MPP+所致神经元凋亡并逆转了MPP+所致形态学改变，这些形态学改变包括不健康的形态和严重损伤的大范围神经炎网络(图1a(B)和图1b(B))。此外，凋亡细胞形态Hoechst33342染色结果显示，SU4312和SU5416能够极大地逆转由MPP+所致的细胞核的固缩(图1a(C)和图1b(C))。
实施例4 SU4312和SU5416对MPTP所致神经毒性的体内保护作用实验
为了测定SU4312，SU5416和PTK787/ZK222584的体内神经保护功能，使用斑马鱼作为动物模型。将1dpf斑马鱼胚胎用200μM MPTP和3μM、10μM、30μM的SU4312或0.1μM、0.3μM、1μM的SU5416或PTK787/ZK222584孵育48小时。处理后，用特异性抗体进行免疫染色和用定量PCR对th基因表达进行分析，从而确定斑马鱼脑部多巴胺***的情况。正对照用MPTP和100μM L-丙炔***(L-Deprenyl，一种单胺氧化酶B抑制剂)进行处理。空白对照组不使用任何试剂进行处理。
用酪氨酸羟化酶抗体进行整体免疫染色的具体步骤如下：将斑马鱼于含4％多聚甲醛的PBS中固定5h，室温下对固定样本进行阻断1h(PBST配2％羊血清和0.1％BSA)。使用小鼠单克隆抗酪氨酸羟化酶抗体(美国Millipore)作为第一抗体，和样本一起于4℃培养过夜。第二天，使用PBST洗脱6次，每次洗脱30min，然后，按照Vectastain ABC试剂盒(美国VectorLab)。染色完成后，用3.5％甲基纤维素平置并照相。
用定量PCR对th基因表达进行分析的具体步骤如下：从每个处理组中随机选取30条斑马鱼幼体，使用RNeasy Mini Kit(美国Qiagen)提取总RNA。用SuperScriptTM III First-StrandSynthesis System for RT-PCR(InvitrogenTM，USA)对RNA进行逆转录，得到单链cDNA，然后在ABI 7500实时PCR***(Applied Biosystems)中使用TaqMan Universal PCR Master Mix和2μL TaqMan用于斑马鱼th基因(分析号：Dr03437803_g1，Applied Biosystems，USA)的基因表达分析引物进行实时PCR。斑马鱼的bactin1引物为5’-CAAGATTCCATACCCAGGAAGGA-3’(F)和5’-CAAGATTCCATACCCAGGAAGGA-3’(R)(Applied Biosystems，USA)。
对每个处理组的TH+神经元进行统计分析，三个独立实验每组20个斑马鱼胚胎，数值以所占对照组百分比表示。数据以对照组的相关差异倍数表示(log2)，显著性差异：*p＜0.05及**p＜0.01(ANOVA和Dunnett’s检验)。
如图2a和图2b所示，进行MPTP处理后，斑马鱼间脑多巴胺神经元的数量(图中括号内部位)大量减少(图2a(A)和图2b(A))，th基因表达水平明显下调(图2a(C))。随着浓度的增加，SU4312和SU5416明显地减缓了多巴胺神经元的减少。相比之下，比较例中的PTK787/ZK222584并不能够阻止斑马鱼中MPTP所致多巴胺神经元的凋亡(图2a)。
进行药物处理后，将3dpf斑马鱼幼体用10μM MPTP和特定浓度的SU4312(3μM、10μM、30μM)、SU5416(0.1μM、0.3μM、1μM)或PTK787/ZK222584(0.3μM、1μM、3μM)孵化72h，阳性对照用10μM MPTP20μM L-丙炔***进行处理。收集处理后的斑马鱼幼体，转入96孔板(每孔1条鱼)。通过自动化视频跟踪***(Viewpoint，ZebraLab，LifeSciences)监视游泳行为。96孔板和照相机置于Zebrabox内，对每条鱼的游泳模式进行录制10min，计算10min内的移动总距离。将移动的总距离定义为斑马鱼在每个时间段(10min)中移动的距离(以厘米计算)。三个独立实验中每组12个斑马鱼幼体，取平均值±方差。显著性差异：与正常组比较，##p＜0.01；与MPTP处理组比较*p＜0.05及**p＜0.01(ANOVA和Dunnett’s检验)。
由于多巴胺神经元受损，MPTP显著地改变了斑马鱼的游泳行为。如图2b(C)和图3(A)、图3(B)所示，斑马鱼幼体游动的总距离在与MPTP接触后显著减少。而用SU4312和SU5416处理后，减轻了MPTP所致斑马鱼游泳行为缺陷，相比之下，比较例中的PTK787/ZK222584则不能够起到减轻该游泳行为缺陷的作用(图3(A)、图3(B))。
实施例5 SU4312和SU5416的神经保护作用与抗血管生成活性的关联性
选择1dpf的Tg(fli1：EGFP)转基因斑马鱼胚胎，在荧光显微镜下对表皮细胞进行观察。分别用不同浓度的SU5416(0.3μM、1μM)、SU4312(1μM、3μM、10μM)和PTK787/ZK222584进行处理48h。进行药物处理后，将斑马鱼从微孔反应板移开，在荧光显微镜(Olympus IX81Motorized Inverted Microscope，Japan)下观察血管的形态变化，该荧光显微镜带有数码照相机(DP controller，Soft Imaging System，Olympus)。用Axiovision 4.2 and Adobe Photoshop 7.0对图像进行分析。
图4显示SU5416，SU4312和PTK787/ZK222584在不同浓度下对斑马鱼幼体节间血管(Intersegmental-Vessel，ISV)形成的抑制效应。与对照组相比，SU4312(1-10μM)和PTK787/ZK222584(0.3-3μM)显示对3dpf的斑马鱼幼体中的ISV形成的不同程度的浓度依赖型抑制，而SU5416(0.3μM，1μM)对ISV无明显抑制，文献报道SU5416浓度在5μM才具有明显抑制斑马鱼ISV效果。事实上，PTK787/ZK222584具有比SU4312和SU5416更有效的抗血管生成活性。然而以上结果表明，SU4312(1-10μM)，SU5416(0.3-1μM)能够阻止斑马鱼的MPTP所致神经元凋亡和运动动缺陷，这些神经保护活性并不与其抗血管生成活性直接相关。
实施例6 SU4312和SU5416对MPP+诱导的神经元中一氧化氮(NO)升高的抑制实验。
用特定浓度的SU5416(20μM、10μM)，SU4312(3μM、10μM、20μM、30μM)处理神经细胞2h，NOS抑制剂ETPU(300μM)和7-NI(3μM、10μM、15μM)作为阳性对照，然后用35μM MPP+诱导神经元内NO产生。用荧光探针DAF-FM diacetate(Molecular Probes，Invitrogen Inc.)检测细胞内NO含量，荧光酶标仪定量分析。数据取三个相互独立的试验的平均值±方差。显著性差异：与MPP+单独处理组比较**p＜0.01(ANOVA and Dunnett’s检验)。
如图5所示，MPP+能明显刺激神经细胞内NO的产生，SU5416或SU4312预处理细胞能剂量依赖地显著抑制MPP+刺激的NO产生。
实施例7 SU4312和SU5416在体外对一氧化氮合成酶(NOS)的抑制实验。
在体外NOS酶活性抑制实验中，纯化的人nNOS，eNOS和iNOS购自于Alexis Biochemicals(Lausen，Switzerland)。根据Calbiochem试剂盒的操作指南，NOS活性用监测L-[3H]arginine转化成[3H]citrulline来测定。试管中的反应体系包含40μl混合物，25 mM Tris-Cl at pH 7.4，3μMtetrahydrobiopterin，1μM FAD，1μM FMN，1mM NADPH，0.6mM CaCl2，0.1μM calmodulin，2.5μg纯的NOS酶，5μl L-[3H]arginine(Perkin Elmer，Waltham，MA，USA)和不同浓度的SU4312(10μM、20μM)或SU5416(10μM)。反应在22℃孵育45min，通过加入400μl of终止缓冲液(50mM HEPES，pH 5.5，and 5mM EDTA)来终止nNOS和eNOS反应或通过加热试管来终止iNOS反应。用液体闪烁计数仪来定量换算NOS抑制活性。
下表表示的是SU4312在体外对nNOS的选择性抑制作用。利用试剂盒中纯化的重组人nNOS，iNOS和eNOS来检测SU4312对它们活性的抑制效果。用Sigma Plot 9.0计算7个稀释浓度下SU4312分别对每种NOS的抑制作用，计算IC50.实验中每个浓度做2个复孔，重复3次。(N.D.(not detectable)表明PTK787在达到1mM浓度时任对3种NO合成酶(NOS)均无明显抑制作用。)
如图6(A)，图6(B)和上表所示，SU4312和SU5416能够非竞争地，选择性阻断nNOS活性。其中，SU4312对nNOS抑制作用IC50约为19μM，对iNOS抑制作用IC50约为390μM，在1mM浓度以下对eNOS无明显抑制作用。而SU5416则浓度依赖地抑制nNOS酶活性，IC50＝22.7±1.3μM(图6(C))，其能够抑制nNOS活性和/或降低nNOS的蛋白表达。
实施例8 蛋白印迹法(Western blotting)检测神经细胞内nNOS蛋白表达实验。
用20μM SU5416预处理神经细胞2h，然后用35μM MPP+处理细胞。神经元细胞经药物处理后，用RIPA裂解液裂解提取蛋白。用SDS聚丙烯酰胺凝胶电泳分离蛋白，将分离的蛋白转移到PVDF膜。然后用5％脱脂奶粉封闭PVFD膜，用抗nNOS一抗4℃反应过夜，洗涤3次后，用HRP-二抗室温反应1h，洗涤3次后，用ECL试剂盒(Amersham Bioscience，Aylesbury，UK)检测蛋白条带，用Bio-Rad自动凝胶成像***拍照，分析条带光密度计算蛋白表达水平。重复3次独立实验求平均值。
如图7所示，MPP+能明显刺激CGNs细胞内nNOS蛋白表达上调，在8-24h内nNOS表达随时间延长逐渐下降，而当SU5416预处理细胞2h，在MPP+刺激的不同时间点均能显著降低nNOS表达上调。
实施例9 SU4312的透血脑屏障实验。
使用HPLC方法检测大鼠脑匀浆中的SU4312。雄性无特异病原Sprague-Dawley(SD)大鼠(250±50g)通过腹腔内注射浓度为12mg/kg的SU4312。分别于给药后15min、30min和1h收集心脏血液，并用PBS/肝素(1000U/L)进行心脏灌流。用断头法杀死大鼠，并使脑组织均质化。用乙酸乙酰酯提取脑组织匀浆中的SU4312并将其重新溶解于甲醇中。用带紫外线-可见光检测器的Agilent 1200 Series HPLC和Agilent ZORBAX XDB-C18柱(2.1×50mm，3.5μm，Agilent Technologies Inc.，CA，USA)进行测定SU4312的浓度，流动速率为0.8ml/min。用Agilent ChemStation(Agilent Technologies Inc.，CA，USA)对检测器输出的数据进行整合和分析。
如图8A所示，SU4312的顺式和反式同分异构体的保留时间为分别9.4min和11.4min。图8C-E表明经SU4312处理的大鼠脑匀浆样品的色谱图(腹腔内注射，12mg/kg，15、30和60min)。即使在腹腔内给药15min后，大鼠脑内的SU4312仍然能够被检测到，这表明SU4312能够很好地通过血脑屏障并到达大脑。
实施例10 SU4312的使用安全性
在光照下，SU4312能够在其顺式异构体和反式异构体之间自由互转。
SU4312的顺式异构体 SU4312的反式异构体
经试验证明，它们均能够选择性地抑制VEGFR-2(顺式异构体IC50值为0.8μM，反式异构体IC50值为5.2μM)。以上研究结果表明，在原代神经元培养和斑马鱼中，SU4312即使是在30μM条件下也不会导所致任何神经毒性。有文献报道，SU4312处理(每5天给药3mg，连续12周)，也不会损害啮齿类动物的视黄醛光感受体或神经节细胞，从而确保SU4312在作为神经保护药物使用是安全无毒的。
Claims (1)
1. 吲哚酮类化合物在制备通过抑制神经元一氧化氮合酶活性而治疗神经退化紊乱疾病的神经保护药物中的应用，其中所述的吲哚酮类化合物选自以下化合物、它们的旋光异构体以及它们与药学上可接受酸的加成盐:
3-(3,′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2。
2. 如权利要求1所述的应用，其特征在于：所述吲哚酮类化合物用于治疗帕金森病。
3. 如权利要求1所述的应用，其特征在于：所述3-(3,′5′-二甲基吡咯-2′-甲烯)1-甲基二氢吲哚酮-2、其旋光异构体或药学上可接受酸的加成盐的浓度为0.3-30 μM。
4. 如权利要求1所述的应用，其特征在于：所述的药物为片剂、颗粒剂、针剂、粉剂、胶囊。
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110327436.8A CN102499917B (zh) | 2011-10-25 | 2011-10-25 | 吲哚酮类化合物在制备神经保护药物中的应用 |
| US14/354,137 US9486440B2 (en) | 2011-10-25 | 2012-10-25 | Uses of indole-ketones or indolidones as neuro-protective drugs |
| PCT/IB2012/055891 WO2013061279A1 (en) | 2011-10-25 | 2012-10-25 | Uses of indole-ketones or indolidones as neuro-protective drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110327436.8A CN102499917B (zh) | 2011-10-25 | 2011-10-25 | 吲哚酮类化合物在制备神经保护药物中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102499917A CN102499917A (zh) | 2012-06-20 |
| CN102499917B true CN102499917B (zh) | 2014-12-17 |
Family
ID=46212095
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110327436.8A Expired - Fee Related CN102499917B (zh) | 2011-10-25 | 2011-10-25 | 吲哚酮类化合物在制备神经保护药物中的应用 |
Country Status (3)
| Country | Link |
|---|---|
| US (1) | US9486440B2 (zh) |
| CN (1) | CN102499917B (zh) |
| WO (1) | WO2013061279A1 (zh) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102499917B (zh) * | 2011-10-25 | 2014-12-17 | 澳门大学 | 吲哚酮类化合物在制备神经保护药物中的应用 |
| US20180311236A1 (en) * | 2015-10-28 | 2018-11-01 | Ab Science | Use of masitinib and other mast cell inhibitors for treatment of parkinson's disease |
| WO2018203559A1 (ja) * | 2017-05-02 | 2018-11-08 | 国立大学法人京都大学 | ポリグルタミン病に関する医薬組成物 |
| CN113891711A (zh) * | 2019-03-11 | 2022-01-04 | 复旦大学 | 用于治疗神经退行性疾病的化合物 |
| CN112569243A (zh) * | 2019-09-30 | 2021-03-30 | 神农医药科技有限公司 | 制备治疗阿尔茨海默病的药物 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010017541A2 (en) * | 2008-08-08 | 2010-02-11 | The Johns Hopkins University | Compositions and methods for treatment of neurodegenerative disease |
| WO2011057204A2 (en) * | 2009-11-06 | 2011-05-12 | The Johns Hopkins University | Lrrk2-mediated neuronal toxicity |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ME00415B (me) | 2000-02-15 | 2011-10-10 | Pharmacia & Upjohn Co Llc | Pirol supstituisani 2-indol protein kinazni inhibitori |
| CA2633069A1 (en) | 2005-12-23 | 2007-09-07 | Centre National De La Recherche Scientifique (Cnrs) | New 3'-, 7-substituted indirubins and their applications |
| CN101023944A (zh) | 2006-02-23 | 2007-08-29 | 黎明涛 | 靛玉红衍生物在制备治疗神经退行性变疾病药物中的应用 |
| CN103945696B (zh) | 2011-04-08 | 2016-06-29 | 贝达医药公司 | 新颖吲哚啉酮蛋白激酶抑制剂 |
| CN102499917B (zh) * | 2011-10-25 | 2014-12-17 | 澳门大学 | 吲哚酮类化合物在制备神经保护药物中的应用 |
-
2011
- 2011-10-25 CN CN201110327436.8A patent/CN102499917B/zh not_active Expired - Fee Related
-
2012
- 2012-10-25 US US14/354,137 patent/US9486440B2/en active Active
- 2012-10-25 WO PCT/IB2012/055891 patent/WO2013061279A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010017541A2 (en) * | 2008-08-08 | 2010-02-11 | The Johns Hopkins University | Compositions and methods for treatment of neurodegenerative disease |
| WO2011057204A2 (en) * | 2009-11-06 | 2011-05-12 | The Johns Hopkins University | Lrrk2-mediated neuronal toxicity |
Non-Patent Citations (1)
| Title |
|---|
| 血管内皮生长因子与中枢神经***疾病及其作用机制;牛小莉等;《临床荟萃》;20100705;第25卷(第13期);参见第1190-1193页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| US9486440B2 (en) | 2016-11-08 |
| WO2013061279A1 (en) | 2013-05-02 |
| US20140271480A1 (en) | 2014-09-18 |
| CN102499917A (zh) | 2012-06-20 |
| WO2013061279A4 (en) | 2013-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hossain et al. | Melatonin in Alzheimer’s disease: a latent endogenous regulator of neurogenesis to mitigate Alzheimer’s neuropathology | |
| US6232326B1 (en) | Treatment for schizophrenia and other dopamine system dysfunctions | |
| CN102499917B (zh) | 吲哚酮类化合物在制备神经保护药物中的应用 | |
| Shekarian et al. | The protective and therapeutic effects of vinpocetine, a PDE1 inhibitor, on oxidative stress and learning and memory impairment induced by an intracerebroventricular (ICV) injection of amyloid beta (aβ) peptide | |
| Huot et al. | Serotonergic approaches in Parkinson’s disease: translational perspectives, an update | |
| JP5605659B2 (ja) | 細胞保護剤 | |
| Kumar et al. | Caffeic acid phenethyl ester (CAPE) prevents development of STZ-ICV induced dementia in rats | |
| Shimizu et al. | Role of cortical and striatal 5-HT1A receptors in alleviating antipsychotic-induced extrapyramidal disorders | |
| Ye et al. | Mechanisms of acupuncture on vascular dementia—a review of animal studies | |
| Yuly Bersudsky et al. | The biology of tryptophan depletion and mood disorders | |
| Huot et al. | Regulation of cortical and striatal 5-HT1A receptors in the MPTP-lesioned macaque | |
| Leclercq et al. | Anticonvulsant and antiepileptogenic effects of system xc− inactivation in chronic epilepsy models | |
| EA036438B1 (ru) | Применение производных пиридазина для предупреждения или лечения атаксического синдрома | |
| CN113967250A (zh) | 乳铁蛋白在预防治疗阿尔兹海默症中的应用 | |
| LeDoux | Movement disorders: Genetics and models | |
| Swerdlow et al. | Neurochemical analysis of rat strain differences in the startle gating-disruptive effects of dopamine agonists | |
| Morin et al. | Contribution of brain serotonin subtype 1B receptors in levodopa-induced motor complications | |
| Chagniel et al. | Striatal inhibition of calpains prevents levodopa-induced neurochemical changes and abnormal involuntary movements in the hemiparkinsonian rat model | |
| Vidyadhara et al. | Admixing MPTP-resistant and MPTP-vulnerable mice enhances striatal field potentials and calbindin-D28K expression to avert motor behaviour deficits | |
| Smith et al. | Pharmacological modulation of amphetamine-induced dyskinesia in transplanted hemi-parkinsonian rats | |
| Haik et al. | 7-nitroindazole attenuates 6-hydroxydopamine-induced spatial learning deficits and dopamine neuron loss in a presymptomatic animal model of Parkinson's disease. | |
| Ahnaou et al. | Off-target potential of AMN082 on sleep EEG and related physiological variables: Evidence from mGluR7 (−/−) mice | |
| Huang et al. | Cordycepin improved the cognitive function through regulating adenosine A2A receptors in MPTP induced Parkinson's disease mice model | |
| Perreault et al. | The atypical dopamine receptor agonist SKF 83959 enhances hippocampal and prefrontal cortical neuronal network activity in a rat model of cognitive dysfunction | |
| Laloux et al. | The effects of serotoninergic, noradrenergic, cholinergic and dopaminergic drugs on vigilance states in MPTP-treated mice |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: THE HONG KONG POLYTECHNIC UNIVERSITY Effective date: 20150519 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20150519 Address after: Chinese Macao Taipa Xu Risheng Yin public road Patentee after: Macao University of China Patentee after: Hong Kong Polytech University Address before: Chinese Macao Taipa Xu Risheng Yin public road Patentee before: Macao University of China |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141217 Termination date: 20211025 |