KR20050031499A - Composition for preventing and treating brain ischemic disease - Google Patents
Composition for preventing and treating brain ischemic disease Download PDFInfo
- Publication number
- KR20050031499A KR20050031499A KR1020030067647A KR20030067647A KR20050031499A KR 20050031499 A KR20050031499 A KR 20050031499A KR 1020030067647 A KR1020030067647 A KR 1020030067647A KR 20030067647 A KR20030067647 A KR 20030067647A KR 20050031499 A KR20050031499 A KR 20050031499A
- Authority
- KR
- South Korea
- Prior art keywords
- cerebral
- group
- ischemia
- hydroxybenzyl
- induction
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 16
- 201000006474 Brain Ischemia Diseases 0.000 title abstract description 40
- BVJSUAQZOZWCKN-UHFFFAOYSA-N p-hydroxybenzyl alcohol Chemical compound OCC1=CC=C(O)C=C1 BVJSUAQZOZWCKN-UHFFFAOYSA-N 0.000 claims abstract description 70
- 206010008118 cerebral infarction Diseases 0.000 claims abstract description 43
- 230000002490 cerebral effect Effects 0.000 claims abstract description 27
- 208000023589 ischemic disease Diseases 0.000 claims abstract description 26
- -1 4-hydroxybenzyl aldehyde Chemical class 0.000 claims abstract description 23
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 claims abstract description 21
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 claims abstract description 21
- 235000012141 vanillin Nutrition 0.000 claims abstract description 21
- 150000001875 compounds Chemical class 0.000 claims abstract description 19
- 208000006011 Stroke Diseases 0.000 claims abstract description 5
- 208000026106 cerebrovascular disease Diseases 0.000 claims abstract description 5
- 206010008111 Cerebral haemorrhage Diseases 0.000 claims abstract description 3
- 208000018737 Parkinson disease Diseases 0.000 claims abstract description 3
- 206010015037 epilepsy Diseases 0.000 claims abstract description 3
- 238000011282 treatment Methods 0.000 claims description 14
- 230000002265 prevention Effects 0.000 claims description 7
- 206010008120 Cerebral ischaemia Diseases 0.000 abstract description 39
- 210000001320 hippocampus Anatomy 0.000 abstract description 10
- 206010008190 Cerebrovascular accident Diseases 0.000 abstract 1
- 230000006907 apoptotic process Effects 0.000 abstract 1
- 208000020658 intracerebral hemorrhage Diseases 0.000 abstract 1
- 230000006698 induction Effects 0.000 description 34
- 208000028867 ischemia Diseases 0.000 description 28
- 210000001519 tissue Anatomy 0.000 description 22
- 230000000302 ischemic effect Effects 0.000 description 20
- 210000004027 cell Anatomy 0.000 description 16
- 239000000243 solution Substances 0.000 description 14
- 230000000971 hippocampal effect Effects 0.000 description 13
- 230000000694 effects Effects 0.000 description 11
- 108010060511 4-Aminobutyrate Transaminase Proteins 0.000 description 10
- 102100035923 4-aminobutyrate aminotransferase, mitochondrial Human genes 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 210000002569 neuron Anatomy 0.000 description 9
- 241000699684 Meriones unguiculatus Species 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- 102000004868 N-Methyl-D-Aspartate Receptors Human genes 0.000 description 8
- 108090001041 N-Methyl-D-Aspartate Receptors Proteins 0.000 description 8
- ZHAFUINZIZIXFC-UHFFFAOYSA-N [9-(dimethylamino)-10-methylbenzo[a]phenoxazin-5-ylidene]azanium;chloride Chemical compound [Cl-].O1C2=CC(=[NH2+])C3=CC=CC=C3C2=NC2=C1C=C(N(C)C)C(C)=C2 ZHAFUINZIZIXFC-UHFFFAOYSA-N 0.000 description 8
- 210000004556 brain Anatomy 0.000 description 8
- 239000011575 calcium Substances 0.000 description 8
- 230000005779 cell damage Effects 0.000 description 8
- 208000037887 cell injury Diseases 0.000 description 8
- 125000003143 4-hydroxybenzyl group Chemical group [H]C([*])([H])C1=C([H])C([H])=C(O[H])C([H])=C1[H] 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 6
- 229910052791 calcium Inorganic materials 0.000 description 6
- 230000007246 mechanism Effects 0.000 description 6
- 238000001000 micrograph Methods 0.000 description 6
- 230000003961 neuronal insult Effects 0.000 description 6
- 239000002953 phosphate buffered saline Substances 0.000 description 6
- 230000010410 reperfusion Effects 0.000 description 6
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 5
- 230000004941 influx Effects 0.000 description 5
- 230000016273 neuron death Effects 0.000 description 5
- 230000036542 oxidative stress Effects 0.000 description 5
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 230000030833 cell death Effects 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 4
- 229930195712 glutamate Natural products 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000010412 perfusion Effects 0.000 description 4
- 239000003755 preservative agent Substances 0.000 description 4
- 150000003254 radicals Chemical class 0.000 description 4
- 229940124597 therapeutic agent Drugs 0.000 description 4
- UUDAMDVQRQNNHZ-UHFFFAOYSA-N (S)-AMPA Chemical compound CC=1ONC(=O)C=1CC(N)C(O)=O UUDAMDVQRQNNHZ-UHFFFAOYSA-N 0.000 description 3
- HCAJQHYUCKICQH-VPENINKCSA-N 8-Oxo-7,8-dihydro-2'-deoxyguanosine Chemical compound C1=2NC(N)=NC(=O)C=2NC(=O)N1[C@H]1C[C@H](O)[C@@H](CO)O1 HCAJQHYUCKICQH-VPENINKCSA-N 0.000 description 3
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 3
- 241000283707 Capra Species 0.000 description 3
- 230000005778 DNA damage Effects 0.000 description 3
- 231100000277 DNA damage Toxicity 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 229920002472 Starch Chemical class 0.000 description 3
- 229920004890 Triton X-100 Polymers 0.000 description 3
- 239000013504 Triton X-100 Substances 0.000 description 3
- VREFGVBLTWBCJP-UHFFFAOYSA-N alprazolam Chemical compound C12=CC(Cl)=CC=C2N2C(C)=NN=C2CN=C1C1=CC=CC=C1 VREFGVBLTWBCJP-UHFFFAOYSA-N 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000003111 delayed effect Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000008107 starch Chemical class 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 238000001356 surgical procedure Methods 0.000 description 3
- 206010002329 Aneurysm Diseases 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 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
- HOKKHZGPKSLGJE-GSVOUGTGSA-N N-Methyl-D-aspartic acid Chemical compound CN[C@@H](C(O)=O)CC(O)=O HOKKHZGPKSLGJE-GSVOUGTGSA-N 0.000 description 2
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 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 2
- 102000003978 Tissue Plasminogen Activator Human genes 0.000 description 2
- 108090000373 Tissue Plasminogen Activator Proteins 0.000 description 2
- 108090000340 Transaminases Proteins 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 230000036760 body temperature Effects 0.000 description 2
- 210000001715 carotid artery Anatomy 0.000 description 2
- 210000001168 carotid artery common Anatomy 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 239000001913 cellulose Chemical class 0.000 description 2
- 229920002678 cellulose Chemical class 0.000 description 2
- OSASVXMJTNOKOY-UHFFFAOYSA-N chlorobutanol Chemical compound CC(C)(O)C(Cl)(Cl)Cl OSASVXMJTNOKOY-UHFFFAOYSA-N 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 238000002695 general anesthesia Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 210000001879 hippocampal ca1 region Anatomy 0.000 description 2
- 239000007972 injectable composition Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000008101 lactose 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
- 238000005259 measurement Methods 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- LXCFILQKKLGQFO-UHFFFAOYSA-N p-hydroxybenzoic acid methyl ester Natural products COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229960000187 tissue plasminogen activator Drugs 0.000 description 2
- 102000014898 transaminase activity proteins Human genes 0.000 description 2
- 201000010875 transient cerebral ischemia Diseases 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- YKBGVTZYEHREMT-KVQBGUIXSA-N 2'-deoxyguanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](CO)O1 YKBGVTZYEHREMT-KVQBGUIXSA-N 0.000 description 1
- YKBGVTZYEHREMT-UHFFFAOYSA-N 2'-deoxyguanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1CC(O)C(CO)O1 YKBGVTZYEHREMT-UHFFFAOYSA-N 0.000 description 1
- KJDSORYAHBAGPP-UHFFFAOYSA-N 4-(3,4-diaminophenyl)benzene-1,2-diamine;hydron;tetrachloride Chemical compound Cl.Cl.Cl.Cl.C1=C(N)C(N)=CC=C1C1=CC=C(N)C(N)=C1 KJDSORYAHBAGPP-UHFFFAOYSA-N 0.000 description 1
- 235000007173 Abies balsamea Nutrition 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 206010001497 Agitation Diseases 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
- 241000416162 Astragalus gummifer Species 0.000 description 1
- 238000012935 Averaging Methods 0.000 description 1
- 239000005528 B01AC05 - Ticlopidine Substances 0.000 description 1
- 239000004857 Balsam Substances 0.000 description 1
- 208000014644 Brain disease Diseases 0.000 description 1
- 206010048962 Brain oedema Diseases 0.000 description 1
- 101710155556 Calcium-dependent protease Proteins 0.000 description 1
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- 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 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 229940123457 Free radical scavenger Drugs 0.000 description 1
- 208000018522 Gastrointestinal disease Diseases 0.000 description 1
- 102000018899 Glutamate Receptors Human genes 0.000 description 1
- 108010027915 Glutamate Receptors Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 208000032843 Hemorrhage Diseases 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
- 241000282412 Homo Species 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 244000018716 Impatiens biflora Species 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102000016193 Metabotropic glutamate receptors Human genes 0.000 description 1
- 108010010914 Metabotropic glutamate receptors Proteins 0.000 description 1
- HOKKHZGPKSLGJE-UHFFFAOYSA-N N-methyl-D-aspartic acid Natural products CNC(C(O)=O)CC(O)=O HOKKHZGPKSLGJE-UHFFFAOYSA-N 0.000 description 1
- 206010029719 Nonspecific reaction Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- 229920001615 Tragacanth Polymers 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 1
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000009056 active transport Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000010419 agar Nutrition 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- SNAAJJQQZSMGQD-UHFFFAOYSA-N aluminum magnesium Chemical compound [Mg].[Al] SNAAJJQQZSMGQD-UHFFFAOYSA-N 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000004019 antithrombin Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 229960000686 benzalkonium chloride Drugs 0.000 description 1
- 235000019445 benzyl alcohol Nutrition 0.000 description 1
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 208000006752 brain edema Diseases 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 230000009460 calcium influx Effects 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 108010079785 calpain inhibitors Proteins 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229960004926 chlorobutanol Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- RRGUKTPIGVIEKM-UHFFFAOYSA-N cilostazol Chemical compound C=1C=C2NC(=O)CCC2=CC=1OCCCCC1=NN=NN1C1CCCCC1 RRGUKTPIGVIEKM-UHFFFAOYSA-N 0.000 description 1
- 229960004588 cilostazol Drugs 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- VGONTNSXDCQUGY-UHFFFAOYSA-N desoxyinosine Natural products C1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 VGONTNSXDCQUGY-UHFFFAOYSA-N 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- KAQKFAOMNZTLHT-VVUHWYTRSA-N epoprostenol Chemical compound O1C(=CCCCC(O)=O)C[C@@H]2[C@@H](/C=C/[C@@H](O)CCCCC)[C@H](O)C[C@@H]21 KAQKFAOMNZTLHT-VVUHWYTRSA-N 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000003825 glutamate receptor antagonist Substances 0.000 description 1
- 150000002334 glycols Chemical class 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 229940029575 guanosine Drugs 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 239000000644 isotonic solution Substances 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000028161 membrane depolarization Effects 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 239000000236 nitric oxide synthase inhibitor Substances 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000006072 paste Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 229940021222 peritoneal dialysis isotonic solution Drugs 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000000106 platelet aggregation inhibitor Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920006316 polyvinylpyrrolidine Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 210000001176 projection neuron Anatomy 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 210000002763 pyramidal cell Anatomy 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 210000001525 retina Anatomy 0.000 description 1
- CSYSULGPHGCBQD-UHFFFAOYSA-N s-ethylisothiouronium diethylphosphate Chemical compound CCSC(N)=N.CCOP(O)(=O)OCC CSYSULGPHGCBQD-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 229940079827 sodium hydrogen sulfite Drugs 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 229940001482 sodium sulfite Drugs 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- AWLILQARPMWUHA-UHFFFAOYSA-M thiopental sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC([S-])=NC1=O AWLILQARPMWUHA-UHFFFAOYSA-M 0.000 description 1
- 229960000340 thiopental sodium Drugs 0.000 description 1
- 229960000103 thrombolytic agent Drugs 0.000 description 1
- 230000002537 thrombolytic effect Effects 0.000 description 1
- PHWBOXQYWZNQIN-UHFFFAOYSA-N ticlopidine Chemical compound ClC1=CC=CC=C1CN1CC(C=CS2)=C2CC1 PHWBOXQYWZNQIN-UHFFFAOYSA-N 0.000 description 1
- 229960005001 ticlopidine Drugs 0.000 description 1
- 239000000196 tragacanth Substances 0.000 description 1
- 235000010487 tragacanth Nutrition 0.000 description 1
- 229940116362 tragacanth Drugs 0.000 description 1
- 238000006276 transfer reaction Methods 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 229960005356 urokinase Drugs 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000000080 wetting agent Substances 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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
-
- 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
- A61K31/11—Aldehydes
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2121/00—Preparations for use in therapy
Landscapes
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Heart & Thoracic Surgery (AREA)
- Cardiology (AREA)
- Vascular Medicine (AREA)
- Urology & Nephrology (AREA)
- General Chemical & Material Sciences (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
Description
본 발명은 뇌허혈성 질환의 예방 및 치료용 조성물에 관한 것으로서, 보다 상세하게는 뇌허혈 및 재관류에 의해 발생되는 신경세포의 손상을 효과적으로 억제하는 뇌허혈성 질환의 예방 및 치료용 조성물에 관한 것이다.The present invention relates to a composition for the prevention and treatment of cerebral ischemic disease, and more particularly, to a composition for the prevention and treatment of cerebral ischemic disease that effectively inhibits the damage of nerve cells caused by cerebral ischemia and reperfusion.
뇌허혈성 질환은 뇌의 혈류가 감소되어 산소와 포도당의 공급이 이루어지지 않아 해마 CA1 영역에 지연성 신경세포사(delayed neuronal death)가 유발되어 발생되는 질환이다(Kirino, Brain Res., 239:57-69, 1982; Petito et al., Neurology, 37:1281-1286, 1987; Pulsinelli et al., Ann. Neurol., 11:491-498, 1982). 또한, 상기 뇌허혈성 뇌질환은 뇌졸중이나 심장마비와 같은 병리학적 상황에서 뇌에 혈액이 공급되지 않음으로써 유발되는 것으로 알려져 있다(Nedergaard, Acta. Neurol. Scand., 77:81-101, 1988; White et al., Neurology, 43:1656-1665, 1993).Cerebral ischemic disease is a disease caused by delayed neuronal death in the hippocampus CA1 region due to decreased blood flow in the brain and lack of oxygen and glucose (Kirino, Brain Res. , 239: 57-). 69, 1982; Petito et al., Neurology , 37: 1281-1286, 1987; Pulsinelli et al., Ann. Neurol. , 11: 491-498, 1982). In addition, the cerebral ischemic brain disease is known to be caused by the lack of blood supply to the brain in pathological conditions such as stroke or heart attack (Nedergaard, Acta. Neurol . Scand. , 77: 81-101, 1988; White; et al., Neurology , 43: 1656-1665, 1993).
일시적인 뇌허혈 및 재관류(reperfusion)에 의해 발생되는 해마 CA1영역에서의 지연성 신경세포사의 원인은 세포내로 칼슘의 과다유입(Hara et al., Brain Res. Bull., 29:659-665, 1992; Nedergaard, Acta. Neurol. Scand., 77:81-101, 1988), 프리 라디컬(free radical) 관련 신경세포 손상 및 글루타메이트-수용체 매개 신경세포 손상이라고 알려져 있다(Won et al., Neurosci. Lett., 301:139-142, 2001). 즉, 뇌허혈이 발생되면 신경세포의 탈분극이 일어나고, 시냅스의 글루타메이트가 유리되어 세포외 글루타메이트의 농도가 증가한다. 또한, ATP의 감소로 인해 발생하는 능동수송의 역전은 세포외 글루타메이트의 축적을 가속시킨다. 상기 세포외 글루타메이트의 축적은 NMDA(N-methyl-D-aspartic Acid), AMPA(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid) 및 메타보트로픽 글루타메이트 수용체(metabotropic glutamate receptor)의 연속적인 활성을 유도하여, 칼슘의 세포내 과다 유입을 초래한다(Olney et al., Science, 244:1360-1362, 1989). 상기 칼슘의 세포내 유입은 특히 칼슘 의존성 프로테아제, 리파제 및 모듈레이터의 활동을 촉발시키고, 결국 프리라디컬을 포함하는 세포 독성분자가 생성되어 DNA 및 세포막 등의 세포 구조물을 파괴하여 신경세포사가 발생하는 것으로 알려져 있다.The cause of delayed neuronal death in the hippocampal CA1 region caused by transient cerebral ischemia and reperfusion is caused by the influx of calcium into cells (Hara et al., Brain Res. Bull ., 29: 659-665, 1992; Nedergaard , Acta. Neurol. Scand. , 77: 81-101, 1988), known as free radical related neuronal damage and glutamate-receptor mediated neuronal damage (Won et al., Neurosci. Lett. , 301: 139-142, 2001). That is, when cerebral ischemia occurs, depolarization of neurons occurs, and glutamate of synapses is released, thereby increasing the concentration of extracellular glutamate. In addition, the reversal of active transport resulting from a decrease in ATP accelerates the accumulation of extracellular glutamate. Accumulation of the extracellular glutamate may include N-methyl-D-aspartic acid (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), and metabotropic glutamate receptor. ), Resulting in an over-flow of calcium into cells (Olney et al., Science , 244: 1360-1362, 1989). Intracellular influx of calcium triggers the activity of calcium-dependent proteases, lipases and modulators, and eventually results in the generation of cytotoxic molecules including free radicals, which destroy cell structures such as DNA and membranes, resulting in neuronal death. Known.
현재 임상에서 사용되고 있는 뇌허혈성 질환의 치료제로는 조직 플라스미노겐 액티베이터(TPA) 및 유로키나제(urokinase)와 같은 혈전용해제, 티클로피딘(ticlopidine), 실로스타졸(cilostazole) 및 프로스타사이클린(prostacycline)과 같은 혈소판 억제제, 항트롬빈제, 항응고제, 뇌혈관 확장제, Ca2+-채널 차단제, 뇌부종 억제제 등 다양한 기전들의 약제들이 있다(Sandercock et al., Hosp. Med., 47:731-736, 1992). 그러나, 이들 약제들은 치료시기가 지연된 경우 그 효과가 미약하고, 급성 뇌허혈로 인한 뇌경색의 진행을 효과적으로 차단하지 못하며, 비특이적 출혈, 피브리노겐 용해 및 급성재폐색 등의 부작용을 나타내는 것으로 알려져 있다.Treatments for cerebral ischemic diseases currently being used in the clinic include thrombolytics such as tissue plasminogen activator (TPA) and urokinase, ticlopidine, cilostazole and prostacycline. Drugs of various mechanisms include platelet inhibitors, antithrombin agents, anticoagulants, cerebrovascular agents, Ca 2+ -channel blockers, and brain edema inhibitors (Sandercock et al., Hosp. Med., 47: 731-736, 1992). However, these agents are known to have a weak effect when treatment is delayed, do not effectively block the progression of cerebral infarction due to acute cerebral ischemia, and have side effects such as nonspecific bleeding, fibrinogen lysis, and acute reoccurrence.
최근에는 뇌허혈로 인한 신경세포 손상 기전에 근거하여 비가역적 손상을 차단 또는 억제할 수 있는 새로운 치료제를 개발하기 위해 많은 연구자들이 다각적 접근을 시도하고 있다. 즉, NMDA나 AMPA와 같은 글루타메이트 수용체 길항제를 비롯하여 Ca2+-채널 차단제, Na+-채널 차단제, 프리 라디컬 소거제, 칼페인(calpain) 억제제, 일산화질소 합성효소 억제제 등이 주요 연구 표적이 되고 있다(Leeson PD and Iversen LL, J. Med. Chem., 37:4053-4067, 1994; Muir KW and Lees KR, Stroke, 26:503-513, 1995). 그 결과로 현재 수종의 약제들이 임상시험 단계에 있으나, 신경세포의 손상 기전이 매우 복잡할 뿐만 아니라 약제의 부작용, 뇌조직으로의 침투력 등 여러가지 문제로 인해 아직까지 효과적인 치료제는 개발되어 있지 않은 실정이다. 따라서, 부작용이 없으면서도 기존 치료제의 문제점을 보완할 수 있는 새로운 형태의 뇌허혈성 질환 치료제에 대한 요구가 증가되고 있다.Recently, many researchers have attempted a multi-faceted approach to develop new therapeutics that can block or suppress irreversible damage based on the mechanism of neuronal damage caused by cerebral ischemia. In other words, glutamate receptor antagonists such as NMDA and AMPA, Ca 2+ -channel blockers, Na + -channel blockers, free radical scavengers, calpain inhibitors, nitric oxide synthase inhibitors, etc. Leeson PD and Iversen LL, J. Med. Chem ., 37: 4053-4067, 1994; Muir KW and Lees KR, Stroke, 26: 503-513, 1995). As a result, several drugs are currently in the clinical trial stage, but the mechanism of neuronal damage is very complicated, and due to various problems such as side effects of drugs and penetration into brain tissue, effective treatments have not been developed. . Therefore, there is an increasing demand for a new type of cerebral ischemic disease therapeutic agent that can supplement the problems of existing therapeutic agents without side effects.
한편, 4-하이드록시벤질 알코올(4-hydroxybenzyl alcohol), 4-하이드록시벤질 알데히드(4-hydroxybenzyl aldehyde) 및 바닐린(vanillin)은 모두 DPPH(1,1-diphenyl-2-picryl hydrazyl) 라디컬, 수퍼옥사이드 라디컬(superoxide radical) 및 하이드록실 라디컬(hydroxyl radical)을 효과적으로 제거하는 항산화 효과를 갖는 것으로 알려져 있다(Liu J. and Mori A., Neuropharmacology, 31:1287-1298, 1992; Luo H et al., Hua Hsi I Ko Ta Hwueh pao, 23: 53-56, 1992).Meanwhile, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin are all DPPH (1,1-diphenyl-2-picryl hydrazyl) radicals, It is known to have an antioxidant effect that effectively removes superoxide radicals and hydroxyl radicals (Liu J. and Mori A., Neuropharmacology , 31: 1287-1298, 1992; Luo H et. al., Hua Hsi I Ko Ta Hwueh pao , 23: 53-56, 1992).
하지만, 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린이 뇌허혈성 질환의 예방 및 치료에 효과가 있다는 사실은 아직 알려져 있지 않다. However, it is not yet known that 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin are effective in the prevention and treatment of cerebral ischemic disease.
본 발명자들은 뇌허혈성 질환의 치료 효과가 우수하면서도, 부작용이 없는 물질을 발견하기 위하여 연구하던 중, 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린이 뇌허혈성 질환의 치료 및 예방에 효과를 가져 뇌허혈성 질환의 치료제로 사용될 수 있음을 확인함으로써 본 발명을 완성하였다. The inventors of the present invention are excellent in the treatment of cerebral ischemic disease, and while studying to find a substance having no side effects, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin are effective in the treatment and prevention of cerebral ischemic disease. The present invention was completed by confirming that it can be used as a therapeutic agent for cerebral ischemic disease.
따라서, 본 발명의 목적은 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린으로 이루어진 군에서 선택된 하나 이상의 화합물을 함유하는 뇌허혈성 질환의 예방 및 치료용 조성물을 제공하는 것이다. Accordingly, it is an object of the present invention to provide a composition for the prevention and treatment of cerebral ischemic disease containing at least one compound selected from the group consisting of 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin.
상기와 같은 목적을 달성하기 위하여, 본 발명은 하기 화학식 1의 4-하이드록시벤질 알코올, 하기 화학식 2의 4-하이드록시벤질 알데히드 및 하기 화학식 3의 바닐린으로 이루어진 군에서 선택된 하나 이상의 화합물을 함유하는 뇌허혈성 질환의 예방 및 치료용 조성물을 제공한다. In order to achieve the above object, the present invention comprises at least one compound selected from the group consisting of 4-hydroxybenzyl alcohol of formula (1), 4-hydroxybenzyl aldehyde of formula (2) and vanillin of formula (3) Provided is a composition for preventing and treating cerebral ischemic disease.
<화학식 1><Formula 1>
<화학식 2><Formula 2>
<화학식 3><Formula 3>
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명에서는 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린으로 이루어진 군에서 선택된 하나 이상의 화합물을 함유하는 뇌허혈성 질환의 예방 및 치료용 조성물을 제공하는 것을 특징으로 한다. The present invention is characterized by providing a composition for preventing and treating cerebral ischemic disease containing at least one compound selected from the group consisting of 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin.
상기 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린은 상업적으로 구입하여 사용하거나 당 업계에 공지된 화학적 합성법으로 제조할 수 있다. 또한, 천연, 바람직하게는 식물, 특히 바람직하게는 천마로부터 분리 정제할 수 있다. 천마로부터 본 발명에 따른 화합물들을 분리 정제하는 방법은 당업계에 공지된 방법을 사용할 수 있다(Kiyoshi Hata et al., Yakugaku Zasshi., 83: 606-610, 1963; Kuk Hyun Shin et al., Arch. Pharm. Res., 17: 331-336, 1994). 바람직하게는 천마의 유효성분을 용매 추출한 후 이를 크로마토그래피로 분리 정제함으로써 목적으로 하는 순수한 화합물을 수득할 수 있다.The 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin may be purchased commercially or prepared by chemical synthesis methods known in the art. It is also possible to separate and purify from natural, preferably plants, and particularly preferably chick horses . As a method for separating and purifying compounds according to the present invention from cheonma , a method known in the art may be used (Kiyoshi Hata et al., Yakugaku Zasshi ., 83: 606-610, 1963; Kuk Hyun Shin et al., Arch Pharm.Res ., 17: 331-336, 1994). Preferably, a pure compound of interest can be obtained by solvent extraction of the active ingredient of cheonma and then separation and purification by chromatography.
본 발명의 일 실시예에서는 본 발명에 따른 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린이 뇌허혈성 질환 치료제로 사용될 수 있는지 확인하기 위하여, 총경동맥(common carotid arteries)을 5분간 폐쇄하는 일시적 전뇌허혈 모델을 사용하여 상기 화합물들이 허혈에 의한 신경세포사를 효과적으로 억제하는지 여부를 조사하였다. 이를 위해, 실험 동물들을 가수술군, 대조군, 4-하이드록시벤질 알코올 투여군, 4-하이드록시벤질 알데히드 투여군 및 바닐린 투여군으로 나눈 후, 각 군에 있어서 성별 및 약물 투여시기에 따른 생존 신경세포수를 비교하였다. In one embodiment of the present invention, to determine whether the 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can be used as a therapeutic agent for cerebral ischemic disease, the common carotid arteries are closed for 5 minutes. A transient whole cerebral ischemia model was used to investigate whether the compounds effectively inhibit neuronal death caused by ischemia. To this end, the experimental animals were divided into the manipulator group, the control group, the 4-hydroxybenzyl alcohol-administered group, the 4-hydroxybenzyl aldehyde-administered group, and the vanillin-administered group. It was.
그 결과, 가수술군에서는 신경세포 손상이 거의 발견되지 않은 반면(생존 세포수 100%), 대조군에서는 심각한 세포손상이 관찰되었다(생존 세포수 11.7%)(도 1참조). 한편, 본 발명에 따른 화합물들을 투여한 경우, 화합물의 종류, 동물의 성별 및 투여시기에 따라 약간의 차이가 있었지만, 생존 세포수는 대조군보다 상당히 많았다(도 2a 내지 4b 참조). 본 발명에 따른 화합물을 투여한 군들의 생존 세포수는 수컷보다 암컷에서 많았고, 허혈유발 전에 투여한 경우가 허혈유발 후에 투여한 경우보다 많았으며(바닐린 암컷 투여군 제외), 대략적으로 4-하이드록시벤질 알코올(평균 52.80%), 바닐린(평균 52.13%) 및 4-하이드록시벤질 알데히드(평균 32.55%) 순으로 많았다(도 5 참조).As a result, almost no neuronal damage was found in the singular group (100% viable cells), whereas severe cell damage was observed in the control group (11.7% viable cells) (see FIG. 1). On the other hand, when administering the compounds according to the present invention, there was a slight difference depending on the type of compound, the sex of the animal and the time of administration, the number of viable cells was significantly higher than the control (see Fig. 2a to 4b). The number of surviving cells in the groups administered with the compound according to the present invention was higher in females than in males, and the administrations before the ischemic induction were higher than those after the ischemic induction (except in the vanillin female administration group), and approximately 4-hydroxybenzyl. Alcohol (average 52.80%), vanillin (average 52.13%), and 4-hydroxybenzyl aldehyde (average 32.55%) were the highest (see FIG. 5).
상기 결과는 본 발명에 따른 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린이 뇌허혈에 의해 발생하는 해마 부위 신경세포사를 효과적으로 억제할 수 있고, 따라서 뇌허혈성 질환의 예방 또는 치료에 탁월한 효과가 있음을 나타낸다.The above results indicate that 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can effectively inhibit hippocampal neuronal cell death caused by cerebral ischemia, and thus have an excellent effect in preventing or treating cerebral ischemic disease. Indicates that there is.
본 발명의 다른 실시예에서는, 상기 화합물 중에서 뇌허혈에 의한 신경 세포 손상을 억제하는 효과가 가장 좋은 것으로 나타난 4-하이드록시벤질 알코올의 뇌허혈성 질환 치료 효과의 작용기전을 확인하기 위해서, 허혈유발 후 시간 경과에 따른 NMDA 수용체 유형 1(NMDA receptor type 1), 8-하이드록시-2'-디옥시구아노신(8-hydroxy-2'-deoxyguanosine) 및 GABA 트랜스아미나제(GABA transaminase)의 변화를 각각 조사하였다. 상기 NMDA 수용체 유형 1은 칼슘의 세포내 유입에 관여하는 것으로 알려져 있고, 8-하이드록시-2'-디옥시구아노신은 산화적 스트레스에 의해 DNA가 손상되는 경우 나타나는 것으로 라디칼에 의해 구아노신의 산화에 의해 생성되는 물질로 알려져 있으며, GABA 트랜스아미나제는 GABA의 아미노 전이 반응을 촉진하는 것으로 알려져 있다. In another embodiment of the present invention, in order to confirm the mechanism of action of the treatment effect of cerebral ischemic disease of 4-hydroxybenzyl alcohol showed the best effect of inhibiting neuronal cell damage caused by cerebral ischemia, the time after ischemic induction The changes of NMDA receptor type 1, 8-hydroxy-2'-deoxyguanosine and GABA transaminase, respectively, were investigated. It was. The NMDA receptor type 1 is known to be involved in the intracellular influx of calcium, and 8-hydroxy-2'-dioxyguanosine appears when DNA is damaged by oxidative stress, which is caused by the oxidation of guanosine by radicals. It is known to produce the substance, GABA transaminase is known to promote the amino transfer reaction of GABA.
그 결과, 뇌허혈유발 30분 전에 4-하이드록시벤질 알코올을 복강내로 투여한 경우, 투여하지 않은 경우에 비해 NMDA 수용체 유형 1의 발현량은 변화가 없었고(도 6 참조), 8-하이드록시-2'-디옥시구아노신의 면역반응은 미약하게 감소하였으며(도 7 참조), GABA 트랜스아미나제의 양은 급격하게 증가하였다(도 8 참조). As a result, when 4-hydroxybenzyl alcohol was administered intraperitoneally 30 minutes before the induction of cerebral ischemia, the expression level of NMDA receptor type 1 was not changed compared with the case without administration (see FIG. 6), and 8-hydroxy-2 The immune response of '-deoxyguanosine was slightly reduced (see FIG. 7), and the amount of GABA transaminase increased rapidly (see FIG. 8).
상기 결과는 본 발명에 따른 4-하이드록시벤질 알코올이 허혈시 발생하는 칼슘 유입의 억제에는 거의 영향을 미치지 못하지만, 산화적 스트레스에 의한 DNA 손상을 억제하고, 빠른 GABA 분해에 의해 발생되는 에너지를 CA1 영역의 추체신경세포(pyramidal neuron cell)에 공급하여 세포 생존에 도움을 줌으로써 뇌허혈에 의해 발생되는 세포 손상을 감소시키는 것으로 판단된다. 한편, 본 발명에 따른 4-하이드록시벤질 알데히드 및 바닐린의 뇌허혈성 질환의 치료 기작은 모두 GABA 트랜스아미나제를 억제함으로써(결과 미도시) 신경세포의 과도한 흥분을 막아 세포사를 억제할 수 있는 것으로 생각되며, 상기 4-하이드록시벤질 알코올의 치료 기작과는 상이한 것으로 판단된다. The above results show that 4-hydroxybenzyl alcohol according to the present invention has little effect on the inhibition of calcium influx generated during ischemia, but inhibits DNA damage caused by oxidative stress and reduces the energy generated by fast GABA degradation. It is thought to reduce cell damage caused by cerebral ischemia by supplying pyramidal neuron cells in the region to help cell survival. On the other hand, the mechanism of treatment of cerebral ischemic diseases of 4-hydroxybenzyl aldehyde and vanillin according to the present invention is thought to be able to inhibit cell death by preventing excessive excitability of neurons by inhibiting GABA transaminase (result not shown). And the mechanism of treatment of the 4-hydroxybenzyl alcohol.
본 발명에 따른 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린은 약학적으로 허용되는 담체를 추가로 포함함으로써 공지의 방법으로 제형화된 후 환자에게 투여될 수 있다. 상기 '약학적으로 허용되는'이란 생리학적으로 허용되고 인간에게 투여될 때, 통상적으로 위장 장애, 현기증 등과 같은 알레르기 반응 또는 이와 유사한 반응을 일으키지 않는 조성물을 말한다.4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can be administered to a patient after being formulated in a known manner by further comprising a pharmaceutically acceptable carrier. The term 'pharmaceutically acceptable' refers to a composition which is physiologically acceptable and does not cause an allergic or similar reaction, such as gastrointestinal disorders, dizziness or the like, when administered to humans.
약학적으로 허용되는 담체로는 예컨대, 경구 투여용 담체 또는 비경구 투여용 담체를 추가로 포함할 수 있다. 경구 투여용 담체는 락토스, 전분, 셀룰로스 유도체, 마그네슘 스테아레이트, 스테아르산 등을 포함할 수 있다. 또한, 비경구 투여용 담체는 물, 적합한 오일, 식염수, 수성 글루코스 및 글리콜 등을 포함할 수 있으며, 안정화제 및 보존제를 추가로 포함할 수 있다. 적합한 안정화제로는 아황산수소나트륨, 아황산나트륨 또는 아스코르브산과 같은 항산화제가 있다. 적합한 보존제로는 벤즈알코늄 클로라이드, 메틸- 또는 프로필-파라벤 및 클로로부탄올이 있다. 그 밖의 약학적으로 허용되는 담체로는 다음의 문헌에 기재되어 있는 것을 참고로 할 수 있다(Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995). Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose and glycols, and the like, and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. Other pharmaceutically acceptable carriers may be referred to those described in the following documents (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).
본 발명에 따른 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린은 상술한 바와 같은 약학적으로 허용되는 담체와 혼합하여 공지의 방법에 따라 다양한 비경구 또는 경구 투여용 형태로 제조될 수 있다. 상기 예를 들어, 비경구 투여용 제형의 대표적인 것은 주사용 제형으로 등장성 수용액 또는 현탁액이 바람직하다. 주사용 제형은 적합한 분산제 또는 습윤제 및 현탁화제를 사용하여 당업계에 공지된 기술에 따라 제조할 수 있다. 예를 들면, 각 성분을 식염수 또는 완충액에 용해시켜 주사용으로 제형화될 수 있다. 또한, 경구 투여용 제형으로는 예를 들면 분말, 과립, 정제, 환약, 캡슐 등이 있는데, 이들 제형은 유효성분 이외에 희석제(예 : 락토즈, 덱스트로즈, 수크로즈, 만니톨, 솔비톨, 셀룰로즈 및/ 또는 글리신), 활탁제(예 : 실리카, 탈크, 스테아르산 및 그의 마그네슘 또는 칼슘염 및/ 또는 폴리에틸렌 글리콜)를 포함할 수 있다. 정제는 또한 마그네슘 알루미늄 실리케이트, 전분페이스트, 젤라틴, 트라가칸스, 메틸셀룰로즈, 나트륨 카복시메틸 셀룰로즈 및/또는 폴리비닐피롤리딘과 같은 결합제를 포함할 수 있으며, 경우에 따라 전분, 한천, 알긴산 또는 그의 나트륨 염과 같은 붕해제 또는 비등 혼합물 및/또는 흡수제, 착색제, 향미제, 및 감미제를 포함할 수 있다. 상기 제형은 통상적인 혼합, 과립화 또는 코팅 방법에 의해 제조될 수 있다. 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can be prepared in various parenteral or oral administration forms according to known methods by mixing with a pharmaceutically acceptable carrier as described above. have. For example, typical of parenteral formulations are injectable formulations, preferably aqueous isotonic solutions or suspensions. Injectable formulations may be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component may be formulated for injection by dissolving in saline or buffer. In addition, oral dosage forms include, for example, powders, granules, tablets, pills, capsules, and the like, which include, in addition to the active ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and the like). / Or glycine), a lubricant, such as silica, talc, stearic acid and its magnesium or calcium salt and / or polyethylene glycol. Tablets may also include binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethyl cellulose and / or polyvinylpyrrolidine, optionally starch, agar, alginic acid or its Disintegrants or boiling mixtures such as sodium salts and / or absorbents, colorants, flavors, and sweeteners. The formulations may be prepared by conventional mixing, granulating or coating methods.
상기와 같은 방법으로 제형화된 약학적 조성물은 경구, 경피, 피하, 정맥 또는 근육을 포함한 여러 경로를 통해 투여될 수 있다.Pharmaceutical compositions formulated in such a manner can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular.
상기에서 '유효량'이란 환자에게 투여하였을 때, 뇌허혈성 질환의 예방 또는 치료 효과를 나타내는 화합물의 양을 말한다. 바람직하게는 본 발명에 따른 화합물들의 통상적인 1일 투여량은 40 내지 60 ㎎/체중㎏의 범위이다. 본 발명에 따른 화합물들은 바람직한 투여량 범위 내에서 1회 또는 수회로 분할 투여할 수 있다. 그러나, 본 발명에 따른 화합물들의 투여량은 투여 경로, 투여 대상, 연령, 성별 체중, 개인차 및 질병 상태에 따라 적절히 선택할 수 있다. 상기에서 환자는 인간을 포함하는 포유동물을 말하며 바람직하게는, 뇌허혈성 질환으로 진단된 인간을 말한다. 상기 뇌허혈성 질환으로는 뇌허혈 및 재관류에 의해 유발되는 모든 질환이 포함된다. 예를 들면, 이에 한정되지는 않으나 뇌졸중, 뇌경색증, 뇌출혈, 간질 또는 파킨슨씨병 등이 포함된다.As used herein, the term 'effective amount' refers to an amount of a compound that exhibits an effect of preventing or treating cerebral ischemic disease when administered to a patient. Preferably a typical daily dosage of the compounds according to the invention is in the range of 40 to 60 mg / kg body weight. The compounds according to the invention can be administered once or in divided doses within the preferred dosage range. However, the dosage of the compounds according to the present invention may be appropriately selected depending on the route of administration, subject to administration, age, gender weight, individual difference and disease state. As used herein, a patient refers to a mammal including a human, and preferably a human diagnosed with cerebral ischemic disease. The cerebral ischemic diseases include all diseases caused by cerebral ischemia and reperfusion. For example, but not limited to, stroke, cerebral infarction, cerebral hemorrhage, epilepsy or Parkinson's disease, and the like.
이하, 본 발명을 실시예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail by way of examples.
단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.
<실시예 1><Example 1>
본 발명에 따른 화합물의 약리 효과 확인Confirmation of the pharmacological effect of the compound according to the present invention
1-1) 일시적 전뇌허혈 손상의 유발1-1) Induction of Temporary Whole Cerebral Ischemia Injury
실험동물은 한림대학교 실험 동물 센터(Experimental Animal Center)로부터 분양받은 몽골리언 저빌(Mongolian gerbil; Meriones unguiculatus)의 자손인, 체중 65~75g의 몽골리언 저빌을 사용하였으며, 상기 실험동물을 오전 7시부터 오후 7시까지 빛을 가하는 일정한 명암주기 하에서 23±2 ℃의 온도 및 50±10%의 상대습도에서 14일 동안 사육하였으며, 음식과 물은 자유로이 섭취하게 하였다.The experimental animal was a Mongolian gerbil, weighing 65-75 g, which is a descendant of the Mongolian gerbil ( Meriones unguiculatus ), which was distributed from the Hallym University Experimental Animal Center. They were kept for 14 days at 23 ± 2 ° C. and 50 ± 10% relative humidity under constant light and light cycles until 7 pm.
일시적 전뇌허혈을 유발하기 위하여, 67 부피%의 일산화질소(nitrous oxide) 및 33 부피%의 산소로 이루어진 가스에 추가로 2.5 부피%의 이소플루란(isoflurane; 미국 Baxtor사)을 첨가한 혼합가스를 사용하여 일반적인 마취 방법에 의해 전신마취시킨 다음, 몽골리언 저빌의 목 부위의 털을 깎고 소독하였다. 그 다음 목 부위를 절개하여 양쪽 총경동맥(common carotid arteries)을 노출시키고, 비외상성 동맥류 클립(non-traumatic aneurysm clip; 미국 Staelting사)을 이용하여 5분 동안 결찰하여 허혈을 유발시켰다. 허혈을 유발시키는 동안, 검안경(ophthalmoscope)을 이용하여 망막중심동맥(central artery of retina)의 혈액 순환 유무를 관찰함으로써 완전한 총경동맥의 폐쇄를 확인하였으며, 직장내 체온계를 삽입하여 체온을 측정하고 실험동물의 온도에 따라 자동으로 조절되는 온열 패드를 사용함으로써 실험동물의 체온을 정상 체온인 37±0.3℃로 일정하게 유지시켰다. 상기 허혈 유발 후 동맥류 클립을 제거하여 재관류시켰으며, 상기 재관류 여부를 현미경 하에서 직접적으로 관찰하였다. In order to cause transient cerebral ischemia, a gas mixture consisting of 67% by volume of nitric oxide and 33% by volume of oxygen in addition to 2.5% by volume of isoflurane (Baxtor, USA) was added. General anesthesia was performed by a general anesthesia method, followed by hair cutting and disinfection of the neck of the Mongolian gerbil. The neck was then incised to expose both common carotid arteries and ligation was induced for 5 minutes using a non-traumatic aneurysm clip (US Staelting). During induction of ischemia, complete obstruction of the total carotid artery was confirmed by observing the blood circulation of the central artery of retina using an ophthalmoscope. The rectal thermometer was inserted to measure body temperature. By using a thermal pad that is automatically adjusted according to the temperature of the animal body temperature was kept constant at 37 ± 0.3 ℃ normal temperature. After the ischemia induction, the aneurysm clip was removed and reperfused, and the reperfusion was observed directly under a microscope.
상기 수술에 의해 허혈이 유발된 암컷, 수컷 몽골리언 저빌에 대하여, 각각 허혈유발 30분전 및 허혈유발 30분후에 10% 트윈 80(0.9% 생리식염수에 녹인) 용액 1 ml에 현탁시킨 4-하이드록시벤질 알코올(Fluka Co.), 4-하이드록시벤질 알데히드 (Junsei Chemical Co.) 및 바닐린(Sigma Chemical Co.)을 각각 40 mg/체중kg 양으로 복강내 투여하여 4-하이드록시벤질 알코올 투여군, 4-하이드록시벤질 알데히드 투여군 및 바닐린 투여군을 제조하였다. 이때 허혈유발 30분전 및 혀혈유발 30분후에 동일한 양의 0.9% 생리식염수에 녹인 10% 트윈 80 용액을 투여한 군을 대조군으로 사용하였다. 다른 그룹의 몽골리언 저빌에 대하여, 총경동맥을 결찰하지 않아 허혈을 유발하지 않았다는 점을 제외하고는 모든 과정이 동일한 가짜(sham)수술을 수행하였다(가수술군). 상기와 같이 상이하게 처리한 동물 그룹을 하기 표 1에 정리하였다. 4-hydroxy was suspended in 1 ml of 10% Tween 80 (dissolved in 0.9% saline solution) 30 minutes before ischemia and 30 minutes after ischemia induction for female and male Mongolian gerbils induced by ischemia. 4-hydroxybenzyl alcohol administration group, benzyl alcohol (Fluka Co.), 4-hydroxybenzyl aldehyde (Junsei Chemical Co.) and vanillin (Sigma Chemical Co.) intraperitoneally administered in an amount of 40 mg / kg body weight, respectively, 4 The hydroxybenzyl aldehyde administration group and the vanillin administration group were prepared. At this time, the group administered with the 10% Tween 80 solution dissolved in the same amount of 0.9% physiological saline 30 minutes before ischemic induction and 30 minutes after tongue induction was used as a control. For another group of Mongolian gerbils, all procedures were performed on the same sham procedure except that the carotid artery was not ligated and caused ischemia (surgery group). Animal groups treated differently as above are summarized in Table 1 below.
1-2) 신경세포 손상 여부 관찰 및 측정1-2) Observation and measurement of nerve cell damage
상기 동물 그룹을 뇌허혈 유발 수술을 수행한 날로부터 4일 후에 희생시켰다. 모든 몽골리언 저빌들에게 티오펜탈 소듐(유한양행, 한국)을 체중kg 당 각각 40 mg의 용량으로 복강 내 주사하여, 마취시킨 다음 1,000 ㎖ 당 헤파린 1000 IU를 함유한 4℃의 생리식염수를 좌심실로 주입하여 관류 세척하였다. 관류 세척이 끝난 동물은 0.1M 포스페이트 버퍼(0.1 M phosphate buffer; PB, pH 7.4)에 녹인 4% 파라포름알데히드(paraformaldehyde)로 관류 고정을 하였다.The animal group was sacrificed 4 days after the day of cerebral ischemia-induced surgery. All Mongolian gerbils were intraperitoneally injected with thiopental sodium (Yuhan, Korea) at a dose of 40 mg / kg body weight, anesthetized and left ventricular at 4 ° C. containing 1000 IU of heparin per 1,000 ml. Perfusion washed by injection. After perfusion washing, the animals were perfused with 4% paraformaldehyde dissolved in 0.1 M phosphate buffer (PB, pH 7.4).
관류 고정이 끝난 동물을 뼈절단기를 이용하여 머리뼈공간을 열어 뇌를 적출한 다음 동일 고정액에서 4-6 시간 후고정 하였다. 후고정이 끝난 뇌를 0.1M 포스페이트 버퍼에 녹인 30% 슈크로즈 용액에 넣어 바닥에 가라앉을 때까지 담가 놓았다. 이 후 활주식 박절기(sliding microtome, Reichert-Jung, 독일)를 이용하여 30 ㎛ 두께로 잘라 보존액(storing solution)이 들어있는 6 웰 플레이트(6 well plate)에 넣어 4℃에서 보관하였다. 각 조직절편 중에서 해마복합체(hippocampal formation)가 잘 나와있는 조직을 골라 조직에 묻어있는 보존액을 제거한 다음 젤라틴을 입힌 슬라이드에 얹어 말려준다. 이 후 증류수에 잠시 담갔다가 2% 크레실 바이올렛(cresyl violet; 미국 Sigma사) 용액에서 1분간 염색한 다음, 흐르는 수돗물에서 충분히 세척하여 슬라이드에 묻어 있는 과량의 크레실 바이올렛을 제거하였다. After perfusion fixation, the brain was removed by using a bone cutter to extract the brain, and then fixed in the same fixative after 4-6 hours. The post-fixed brains were soaked in 30% sucrose solution dissolved in 0.1 M phosphate buffer and soaked until they sank to the bottom. Thereafter, using a sliding microtome (sliding microtome, Reichert-Jung, Germany) was cut into 30 ㎛ thickness into a 6 well plate containing a storage solution (storing solution) and stored at 4 ℃. Choose a tissue that shows well hippocampal formation among each tissue section, remove the preservatives from the tissue, and dry it on a gelatinized slide. Thereafter, the solution was briefly dipped in distilled water, dyed in 2% cresyl violet (Sigma, USA) solution for 1 minute, and then washed in running tap water to remove excess cresyl violet on the slide.
해마 CA1 손상 여부는 크레실 바이올렛 용액으로 염색된 생존 뉴런을 계수함으로써 결정하였다. 각 그룹의 동물에 대하여, 해마 절편 내의 양측 반구(hemisphere)의 밀리미터당 CA1 뉴런의 평균수를 측정하였다. 신경세포체(neuronal somata) 수의 측정은 컴퓨터에 기초한 CCD 카메라가 장착된 영상 분석 시스템(소프트웨어: Optimas 6.5, 미국 CyberMetrics사)을 사용하여 수행하였다. 각 동물로부터 얻어진 21개 절편으로부터의 염색된 신경세포수를 평균함으로써 세포를 계수하였다. 통계적 유의성을 검증하기 위해 정량 데이터로부터 얻어진 모든 데이터는 원웨이 아노바 테스트(One-way ANOVA test)를 이용하여 분석되었고, 포스트혹(post hoc) 비교를 위하여 본페로니 테스트(Bonferroni's test)를 수행하였다. 유의성은 p값이 0.01 이하일 때로 정하였다. Hippocampal CA1 damage was determined by counting viable neurons stained with cresyl violet solution. For each group of animals, the average number of CA1 neurons per millimeter of bilateral hemispheres in hippocampal sections was measured. The measurement of the number of neuronal somata was performed using an image analysis system (software: Optimas 6.5, CyberMetrics, USA) equipped with a computer-based CCD camera. Cells were counted by averaging the number of stained neurons from 21 sections obtained from each animal. All data obtained from quantitative data to verify statistical significance were analyzed using the One-way ANOVA test and Bonferroni's test was performed for post hoc comparison. It was. Significance was determined when p value was 0.01 or less.
각 군의 해마조직 및 그의 CA1 영역을 크레실 바이올렛으로 염색한 현미경 사진을 도 1 내지 도4에 나타내었다. 도 1에 나타낸 바와 같이, 허혈을 유발하지 않은 가수술군에서는 세포 손상이 거의 발견되지 않은 반면(도 1A 및 1C), 대조군에서는 심각한 세포 손상이 있었다(도 1B 및 1D). 한편, 4-하이드록시벤질 알코올 투여군, 4-하이드록시벤질 알데히드 투여군 및 바닐린 투여군의 경우 상기 가수술군에 비해 상대적으로 세포 손상이 심했지만, 대조군에 비해서는 세포 손상이 상당히 감소하였다(도 2a 내지 도 4b). Micrographs of the hippocampal tissue and its CA1 region of each group stained with cresyl violet are shown in FIGS. 1 to 4. As shown in FIG. 1, almost no cell damage was found in the ischaic group that did not induce ischemia (FIGS. 1A and 1C), whereas there was severe cell damage in the control group (FIGS. 1B and 1D). Meanwhile, the 4-hydroxybenzyl alcohol-administered group, 4-hydroxybenzyl aldehyde-administered group, and vanillin-administered group showed more severe cell damage than the hydrolyzate group, but significantly reduced cell damage compared to the control group (FIGS. 2A to 2). 4b).
신경세포 손상이 거의 없는 가수술군(100%)을 기준으로 각 군의 생존 세포수를 계수한 결과를 도 5에 나타내었다. 도 5에 나타낸 바와 같이, 본 발명에 따른 화합물을 투여한 군들의 생존 세포수는 수컷보다 암컷에서 많았고, 허혈유발 전에 투여한 경우가 허혈유발 후에 투여한 경우보다 많았으며(바닐린 투여군 제외), 대략적으로 4-하이드록시벤질 알코올(평균 52.80%), 바닐린(평균 52.13%) 및 4-하이드록시벤질 알데히드(평균 32.55%) 순으로 많았다.5 shows the results of counting the number of viable cells in each group based on the manipulator group (100%) having almost no neuronal damage. As shown in FIG. 5, the number of surviving cells in the groups administered with the compound according to the present invention was higher in females than in males, and more frequently administered before ischemic induction than when administered after ischemic induction (except vanillin-administered group). 4-hydroxybenzyl alcohol (average 52.80%), vanillin (average 52.13%) and 4-hydroxybenzyl aldehyde (average 32.55%).
상기 결과에 의하여, 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린은 뇌허혈 및 재관류에 의해 발생하는 해마 부위 세포의 세포사를 효과적으로 억제함을 확인할 수 있었다. As a result, it was confirmed that 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin effectively inhibit the cell death of hippocampal region cells caused by cerebral ischemia and reperfusion.
<실시예 2><Example 2>
4-하이드록시벤질 알코올의 작용기전 확인Confirmation of functional mechanism of 4-hydroxybenzyl alcohol
본 발명자들은 본 발명의 화합물 중에서 뇌허혈 및 재관류에 의해 발생하는 해마 부위 세포의 세포사를 가장 효과적으로 억제하는 것(도 5 참조)으로 나타난 4-하이드록시벤질 알코올의 작용기전을 확인하기 위해서, 허혈유발 후 시간 경과에 따른 NMDA 수용체 유형 1(NMDA receptor type 1), 8-하이드록시-2'-디옥시구아노신(8-hydroxy-2'-deoxyguanosine) 및 GABA 트랜스아미나제(GABA transaminase)의 변화를 각각 조사하였다. The inventors of the present invention to determine the mechanism of action of 4-hydroxybenzyl alcohol shown to most effectively inhibit the cell death of hippocampal region cells caused by cerebral ischemia and reperfusion among the compounds of the present invention, after ischemic induction Changes in NMDA receptor type 1, 8-hydroxy-2'-deoxyguanosine and GABA transaminase, respectively, over time Investigate.
상기 실시예 1과 동일한 방법으로 뇌허혈 유발 실험 동물을 제조하였다. 상기 뇌허혈을 유발하기 30분 전의 몽골리안 저빌에 4-하이드록시벤질 알코올 40mg/kg을 복강 내로 투여하였다. 이후, 허혈 유발전, 허혈 유발후 30분, 3시간, 6시간, 12시간 및 24시간에서 각각 동물을 희생하여, 상기 실시예에서와 동일한 방법으로 관류 고정을 실시하고, 30 ㎛ 두께로 조직을 잘라 보존액이 들어있는 6 웰 플레이에 넣어 보관하였다. 각 조직절편 중에서 해마가 잘 나와 있는 조직을 골라 0.1M PBS(phosphate buffered saline, pH 7.4)로 10분 동안 3회 세척하여 조직에 묻어 있던 보존액을 제거한 다음 하기의 ABC법을 이용하여 면역 염색을 수행하였다. 즉, 상기 세척이 끝난 조직 절편에 대해 조직 내에 존재하는 내인성 퍼옥시다제(peroxidase)를 제거하기 위해 0.01M PBS 중의 0.3% H2O2를 30분 동안 처리하였고, 비특이적 반응을 방지하기 위하여 5% 표준 산양 혈청(normal goat serum)을 처리하여 30분 동안 반응시켰다. 1차 항체로 각각 마우스 항-NMDA 수용체 유형 1(mouse anti-NMDA receptor type 1: 미국 Chemicon International사), 마우스 항-8-하이드록시-2'-디옥시구아노신(mouse anti-8-hydroxy-2'-deoxyguanosine: 미국 Peninsula사) 및 마우스 항-GABA 트랜스아미나제(mouse anti-GABA transaminase: 한림대 최수영 교수 제공)를 2% 표준 산양 혈청 및 0.1% 트리톤 X-100(Triton X-100)이 함유된 0.1M PBS 용액에 1:100의 비율로 희석하여 상기 처리된 각 조직과 함께 4℃에서 24시간 동안 반응시켰다. 이후 2차 항체로 바이오틴이 부착된 산양-마우스 IgG(biotinylated-goat anti-mouse IgG: 미국 Vector사)를 2% 표준 산양 혈청 및 0.1% 트리톤 X-100(Triton X-100)이 함유된 0.1M PBS 용액에 1:200으로 희석하여 상기 각 조직과 함께 상온에서 2시간 반응시켰으며, 이후 상기 조직을 벡터 키트(미국 Vector사) 용액과 상온에서 1시간 반응시켰다. 상기 각 단계의 반응 후에는 상기 조직을 PBS 용액으로 4차례씩 세척하였다. 항원항체 반응이 끝난 조직은 0.03% 과산화수소수와 0.05% DAB(3,3'-diaminobenzidine tetrachloride: 미국 Sigma사)가 함유된 트리스 버퍼(Tris buffer: pH 7.4) 용액에서 1-2분 동안 발색시킨 다음 세척한 후 젤라틴이 입혀진 슬라이드에 얹어 건조시키고, 통상적인 탈수와 투명화 과정을 거쳐 캐나다 발삼(일본 Kanto사)으로 봉입하였다. 상기 면역조직화학 염색법을 통한 허혈유발 후 시간 경과에 따른 NMDA 수용체 유형 1, 8-하이드록시-2'-디옥시구아노신 및 GABA 트랜스아미나제의 변화를 도 6 내지 8에 각각 나타내었다.A cerebral ischemia-induced experimental animal was prepared in the same manner as in Example 1. 40 mg / kg of 4-hydroxybenzyl alcohol was intraperitoneally administered to Mongolian gerbil 30 minutes before the cerebral ischemia was induced. Thereafter, animals were sacrificed at 30 minutes, 3 hours, 6 hours, 12 hours, and 24 hours before ischemia induction, respectively, and the perfusion fixation was performed in the same manner as in the above example, and the tissue was 30 μm thick. Cut and stored in 6 well play containing the stock solution. Choose a tissue that shows well hippocampus among each tissue section, wash it three times for 10 minutes with 0.1M PBS (phosphate buffered saline, pH 7.4) to remove the preservative solution from the tissue, and then perform immunostaining using the following ABC method. It was. That is, the washed tissue sections were treated with 0.3% H 2 O 2 in 0.01 M PBS for 30 minutes to remove endogenous peroxidase present in the tissue, and 5% to prevent nonspecific reactions. Standard goat serum was treated and reacted for 30 minutes. The primary antibodies were mouse anti-NMDA receptor type 1 (chemicon International, USA) and mouse anti-8-hydroxy-2'-deoxyguanosine (mouse anti-8-hydroxy-) 2'-deoxyguanosine: Peninsula, USA and mouse anti-GABA transaminase (provided by Professor Soo Young Choi, Hallym University) contain 2% standard goat serum and 0.1% Triton X-100 (Triton X-100) The diluted 0.1 M PBS solution was diluted at a ratio of 1: 100 and reacted with each of the treated tissues at 4 ° C. for 24 hours. Afterwards, the biotinylated goat-mouse IgG (Biotinylated-goat Anti-mouse IgG: Vector of USA) was used as a secondary antibody and 0.1M containing 2% standard goat serum and 0.1% Triton X-100. The solution was diluted 1: 200 in PBS solution and reacted with each of the tissues at room temperature for 2 hours, after which the tissue was reacted with the vector kit (US Vector) solution at room temperature for 1 hour. After each reaction, the tissues were washed four times with PBS solution. After the antigen-antibody reaction, the tissue was developed for 1-2 minutes in a solution of Tris buffer (pH 7.4) containing 0.03% hydrogen peroxide solution and 0.05% DAB (3,3'-diaminobenzidine tetrachloride, Sigma, USA). After washing, it was dried on gelatin-coated slides, dried, and then sealed with Canadian balsam (Kanto, Japan) through a conventional dehydration and clarification process. The changes of NMDA receptor type 1, 8-hydroxy-2'-dioxyguanosine and GABA transaminase with time after ischemic induction through the immunohistochemical staining are shown in FIGS. 6 to 8, respectively.
도 6에 도시되어 있는 바와 같이, 허혈 발생시 칼슘의 세포내 유입에 관여하는 것으로 알려져 있는 NMDA 수용체 유형 1의 발현은 뇌허혈 유발 후 경시적인 변화가 미약하였다(도 6). 상기 결과는 본 발명자들이 허혈유발 전에 4-하이드록시벤질 알코올을 투여한 것을 제외하고 동일하게 실험한 이전의 변화양상(Kang et al., J Neurocytol., 30:945-955, 2001)과 일치하는 것이다. 상기 결과에 의하여, 4-하이드록시벤질 알코올의 투여가 NMDA 수용체 유형 1의 발현에 영향을 미치지 않으므로, 4-하이드록시벤질 알코올은 허혈 발생시 일어나는 초기 칼슘의 유입에는 거의 영향을 미치지 않는 것을 알 수 있었다.As shown in FIG. 6, the expression of NMDA receptor type 1, which is known to be involved in the intracellular influx of calcium during ischemia development, has a slight change over time after cerebral ischemia induction (FIG. 6). The results are consistent with previous changes of the same experiments by the present inventors except that 4-hydroxybenzyl alcohol was administered prior to ischemic induction (Kang et al., J Neurocytol ., 30: 945-955, 2001) . will be. The results showed that 4-hydroxybenzyl alcohol did not affect the expression of NMDA receptor type 1, so 4-hydroxybenzyl alcohol had little effect on the influx of early calcium during ischemia. .
한편, 도 7에 도시되어 있는 바와 같이, 산화적 스트레스(oxidative stress)에 의해 DNA가 손상되어 생성되는 산화물질인 8-하이드록시-2'-디옥시구아노신은 뇌허혈 유발 12시간까지 조금씩 증가하였고, 뇌허혈 유발 12시간에서의 면역반응은 허혈유발 전에 비해 150% 정도 증가하였다. 상기 결과는 본 발명자들이 허혈유발 전에 4-하이드록시벤질 알코올을 투여한 것을 제외하고 동일하게 수행한 실험에서 면역반응이 200% 이상 증가했다는 이전의 결과(Won et al., Brain Res., 836: 70-78, 1999)보다 적은 값이다. 상기 결과에 의하여, 4-하이드록시벤질 알코올의 투여가 8-하이드록시-2'-디옥시구아노신의 증가를 억제하는 것을 알 수 있었고, 따라서 허혈 발생 초기에 일어나는 산화적 스트레스에 의한 DNA 손상을 억제하는 것으로 생각된다.On the other hand, as shown in Figure 7, 8-hydroxy-2'-dioxyguanosine, an oxide produced by DNA damage due to oxidative stress (oxidative stress) slightly increased up to 12 hours to induce cerebral ischemia, The immune response at 12 hours of cerebral ischemia was increased by 150% compared to before ischemia induction. The above results indicate that the previous results showed that the immune response increased more than 200% in the same experiments performed by the inventors except that 4-hydroxybenzyl alcohol was administered before ischemic induction (Won et al., Brain Res., 836: 70-78, 1999). The results showed that administration of 4-hydroxybenzyl alcohol inhibited the increase of 8-hydroxy-2'-dioxyguanosine, thus inhibiting DNA damage due to oxidative stress occurring early in ischemia development. I think.
한편, 도 8에 도시되어 있는 바와 같이, GABA 의 아미노 전이반응을 촉진하는 GABA 트랜스아미나제의 경우, 허혈유발후 3시간에 급격히 증가하기 시작하였다. 상기 결과는 본 발명자들이 허혈유발 전에 4-하이드록시벤질 알코올을 투여한 것을 제외하고 동일하게 수행한 실험에서 GABA 트랜스아미나제의 급격한 증가는 허혈유발후 12시간에서 일어났다는 이전의 결과(Kang et al., Neurosci Lett., 294:33-36, 2000)보다 훨씬 빠르다. 상기 결과에 의하여, 4-하이드록시벤질 알코올의 투여는 GABA 트랜스아미나제의 증가를 억제하는 능력을 저하시킴을 알 수 있었고, 따라서 빠른 GABA 분해에 의해 발생되는 에너지를 CA1 영역의 추체신경세포(pyramidal neuron cell)에 공급하여 세포의 생존에 도움을 주는 것으로 생각된다.On the other hand, as shown in Figure 8, in the case of GABA transaminase to promote the aminotransferase reaction of GABA, it began to increase rapidly 3 hours after ischemia induction. The results indicate that the rapid increase of GABA transaminase occurred 12 hours after ischemic induction in the experiments performed by the inventors in the same manner except that 4-hydroxybenzyl alcohol was administered before ischemic induction (Kang et al. , Neurosci Lett., 294: 33-36, 2000). As a result, administration of 4-hydroxybenzyl alcohol lowered the ability to inhibit the increase of GABA transaminase, and thus the energy generated by fast GABA degradation was reduced to pyramidal cells in the CA1 region. It is thought to contribute to the survival of cells by supplying them to neuron cells.
상기 기재한 바와 같이, 4-하이드록시벤질 알코올, 4-하이드록시벤질 알데히드 및 바닐린은 뇌허혈-재관류에 의해 발생하는 해마 부위 세포의 세포사를 억제하는 효과를 갖고 있다. 따라서, 상기 화합물들은 뇌허혈성 질환의 예방 및 치료에 효과적으로 이용될 수 있다. As described above, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin have the effect of inhibiting cell death of hippocampal site cells caused by cerebral ischemia-reperfusion. Thus, the compounds can be effectively used for the prevention and treatment of cerebral ischemic disease.
도 1은 가수술군 및 대조군의 해마조직 및 그의 CA1 영역을 크레실 바이올렛(cresyl violet)으로 염색한 현미경 사진을 나타낸 것이다(A: 가수술군의 해마조직, B: 대조군의 해마조직, C: 가수술군의 해마 CA1 영역, D: 대조군의 해마 CA1 영역). Figure 1 shows a micrograph of the hippocampal tissue and its CA1 region stained with cresyl violet (A: hippocampus tissue of the Singer group, B: hippocampal tissue of the control group, C: Singer surgery group) Hippocampus CA1 region, D: hippocampus CA1 region of the control group).
도 2a 및 도 2b는 4-하이드록시벤질 알코올 투여군의 해마조직 및 그의 CA1 영역을 크레실 바이올렛으로 염색한 현미경 사진을 각각 나타낸 것이다(A: 허혈유발 30분전 투여군(수컷), B: 허혈유발 30분후 투여군(수컷), C: 허혈유발 30분전 투여군(암컷), D: 허혈유발 30분후 투여군(암컷)). 2A and 2B show microscopic photographs of the hippocampal tissue and its CA1 region of the 4-hydroxybenzyl alcohol-administered group with cresyl violet, respectively (A: 30 minutes before ischemia-induced group (male), B: ischemia-induced 30). Post-administration group (male), C: administration group 30 minutes before ischemia-induced (female), D: administration group (female) 30 minutes after ischemia-induced).
도 3a 및 도 3b는 4-하이드록시벤질 알데히드 투여군의 해마조직 및 그의 CA1 영역을 크레실 바이올렛으로 염색한 현미경 사진을 나타낸 것이다(A: 허혈유발 30분전 투여군(수컷), B: 허혈유발 30분후 투여군(수컷), C: 허혈유발 30분전 투여군(암컷), D: 허혈유발 30분후 투여군(암컷)). 3A and 3B show micrographs of the hippocampal tissue of the 4-hydroxybenzyl aldehyde-administered group and its CA1 region stained with cresyl violet (A: 30 minutes before ischemia-induced group (male), B: 30 minutes after ischemia-induced group) Administration group (male), C: administration group 30 minutes before ischemic induction (female), D: administration group (female) 30 minutes after ischemic induction).
도 4a 및 도 4b는 바닐린 투여군의 해마조직 및 그의 CA1 영역을 크레실 바이올렛으로 염색한 현미경 사진을 각각 나타낸 것이다(A: 허혈유발 30분전 투여군(수컷), B: 허혈유발 30분후 투여군(수컷), C: 허혈유발 30분전 투여군(암컷), D: 허혈유발 30분후 투여군(암컷)). 4A and 4B show microscopic photographs of the hippocampus tissue and its CA1 region stained with cresyl violet, respectively (A: 30 minutes before ischemia-induced group (male), B: 30 minutes after ischemia-induced group (male)). C: group administered 30 minutes before ischemic induction (female), D: group administered 30 minutes after ischemic induction (female).
도 5는 각 군의 해마 조직에서 계수된 허혈유발 후 생존 해마세포의 평균수를 가수술군을 기준으로 나타낸 것이다. Figure 5 shows the average number of viable hippocampal cells after ischemic induction in the hippocampus tissue of each group based on the manipulator group.
도 6은 허혈유발 30분전에 4-하이드록시벤질 알코올을 투여한 경우, NMDA 수용체 유형 1(NMDA receptor type 1)에 대한 면역조직염색에 의한 해마 CA1 영역의 현미경 사진이다(A: 뇌허혈전, B; 뇌허혈 후 30분, C: 뇌허혈 후 3시간, D: 뇌허혈 후 6시간, E: 뇌허혈 후 12시간, F: 뇌허혈 후 24시간). Figure 6 is a micrograph of the hippocampus CA1 region by immunohistostaining for NMDA receptor type 1 when 4-hydroxybenzyl alcohol was administered 30 minutes before ischemia induction (A: cerebral ischemia, B). 30 minutes after cerebral ischemia, C: 3 hours after cerebral ischemia, D: 6 hours after cerebral ischemia, E: 12 hours after cerebral ischemia, F: 24 hours after cerebral ischemia).
도 7은 허혈유발 30분전에 4-하이드록시벤질 알코올을 투여한 경우, 8-하이드록시-2'-디옥시구아노신(8-hydroxy-2'-deoxyguanosine)에 대한 면역조직염색에 의한 해마 CA1 영역의 현미경 사진이다(A: 뇌허혈전, B; 뇌허혈 후 30분, C: 뇌허혈 후 3시간, D: 뇌허혈 후 6시간, E: 뇌허혈 후 12시간, F: 뇌허혈 후 24시간). FIG. 7 shows hippocampal CA1 due to immunohistostaining for 8-hydroxy-2'-deoxyguanosine when 4-hydroxybenzyl alcohol was administered 30 minutes before ischemia induction. A micrograph of the area (A: cerebral ischemia, B; 30 minutes after cerebral ischemia, C: 3 hours after cerebral ischemia, D: 6 hours after cerebral ischemia, E: 12 hours after cerebral ischemia, F: 24 hours after cerebral ischemia).
도 8은 허혈유발 30분전에 4-하이드록시벤질 알코올을 투여한 경우, GABA 트랜스아미나제(GABA transaminase)에 대한 면역조직염색에 의한 해마 CA1 영역의 현미경 사진이다(A: 뇌허혈전, B; 뇌허혈 후 30분, C: 뇌허혈 후 3시간, D: 뇌허혈 후 6시간, E: 뇌허혈 후 12시간, F: 뇌허혈 후 24시간). Figure 8 is a micrograph of the hippocampal CA1 region by immunohistostaining for GABA transaminase when 4-hydroxybenzyl alcohol was administered 30 minutes before ischemia induction (A: cerebral ischemia, B; cerebral ischemia). 30 minutes later, C: 3 hours after cerebral ischemia, D: 6 hours after cerebral ischemia, E: 12 hours after cerebral ischemia, F: 24 hours after cerebral ischemia).
Claims (2)
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020030067647A KR20050031499A (en) | 2003-09-30 | 2003-09-30 | Composition for preventing and treating brain ischemic disease |
AU2003282440A AU2003282440A1 (en) | 2003-09-30 | 2003-11-26 | Novel therapeutic use of 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin |
PCT/KR2003/002578 WO2005030189A1 (en) | 2003-09-30 | 2003-11-26 | Novel therapeutic use of 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020030067647A KR20050031499A (en) | 2003-09-30 | 2003-09-30 | Composition for preventing and treating brain ischemic disease |
Publications (1)
Publication Number | Publication Date |
---|---|
KR20050031499A true KR20050031499A (en) | 2005-04-06 |
Family
ID=34386636
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020030067647A KR20050031499A (en) | 2003-09-30 | 2003-09-30 | Composition for preventing and treating brain ischemic disease |
Country Status (3)
Country | Link |
---|---|
KR (1) | KR20050031499A (en) |
AU (1) | AU2003282440A1 (en) |
WO (1) | WO2005030189A1 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010015965A2 (en) * | 2008-08-04 | 2010-02-11 | Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. | Methods and compositions for treating neuronal damage and modulating transient receptor potential channels |
EP2886656A1 (en) | 2013-12-18 | 2015-06-24 | Commissariat A L'energie Atomique Et Aux Energies Alternatives | New enzyme and method for preparing 4-hydroxyl benzyl alcohol and derivatives thereof |
-
2003
- 2003-09-30 KR KR1020030067647A patent/KR20050031499A/en not_active Application Discontinuation
- 2003-11-26 WO PCT/KR2003/002578 patent/WO2005030189A1/en active Application Filing
- 2003-11-26 AU AU2003282440A patent/AU2003282440A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
AU2003282440A1 (en) | 2005-04-14 |
WO2005030189A1 (en) | 2005-04-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Gao et al. | Tau in Alzheimer's disease: mechanisms and therapeutic strategies | |
CN106102737B (en) | Cromoglycic acid derivative and the correlation technique of imaging and treatment | |
EA011074B1 (en) | Hydrazide-containing cftr inhibitor compounds and uses thereof | |
CN109475539B (en) | Treatment of parkinson's disease | |
US9192610B2 (en) | Use of quinazoline derivatives for neurodegenerative diseases | |
US10583105B2 (en) | Compositions and methods for neuroprotection and treatment of neurodegeneration | |
ES2683184T3 (en) | GLyT1 inhibitors for use in the treatment of blood disorders | |
AU2023200727A1 (en) | Methods and compositions for treating psychotic disorders | |
BR112021002692A2 (en) | use of riluzole oral disintegrating tablets for the treatment of diseases | |
WO2014090990A1 (en) | Leukotriene pathway antagonists for the treatment of dementia, cognitive deficits in parkinson's disease and/or learning and memory deficiencies in parkinson's disease | |
CN115996728A (en) | Methods of treating Alzheimer's disease using Rho kinase inhibitors | |
AU2016372396B2 (en) | 2-iminobiotin for use in the treatment of brain cell injury | |
JP2018531961A (en) | Methods and compositions for recovery from stroke | |
TWI472519B (en) | N-butylidenephthalide-containing pharmaceutical composition for treating liver injury and improving liver function | |
KR20050031499A (en) | Composition for preventing and treating brain ischemic disease | |
US8481500B2 (en) | Compounds having neuroprotective properties | |
KR20220076375A (en) | A novel pharmaceutical composition for treating neurodegenerative disease | |
JP7295145B2 (en) | Medicaments and uses thereof for treating neurodegenerative diseases | |
CN116249532A (en) | Methods of treating vascular dementia using Rho kinase inhibitors | |
JP2012184263A (en) | Pyrazolone compounds useful for treatment of cerebrovascular disorder associated with ischemic stroke | |
EP3431468B1 (en) | Use of adiponectin receptor agonists in treating depression, anxiety and neuroinflammation | |
KR20200093976A (en) | Pharmaceutical Composition for Inhibiting Amyloid Beta Deposition Comprising Uncaria Rhynchophylla Extract | |
KR101855087B1 (en) | Chalcone derivatives, optical isomer thereof, or pharmaceutically acceptable salts thereof, and a pharmaceutical composition for preventing or treating mitochondrial disease induced by decrease of oxygen consumption rate comprising the same as an active ingredient | |
CN117599062B (en) | Composition for assisting in improving memory and preventing or treating Alzheimer disease | |
US20240189307A1 (en) | Methods of stabilizing the neuronal proteome against collapse and protecting vascular cells |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A201 | Request for examination | ||
E902 | Notification of reason for refusal | ||
E601 | Decision to refuse application |