CN100526305C - Compound for promoting neurocyte growth - Google Patents
Compound for promoting neurocyte growth Download PDFInfo
- Publication number
- CN100526305C CN100526305C CNB2005100975229A CN200510097522A CN100526305C CN 100526305 C CN100526305 C CN 100526305C CN B2005100975229 A CNB2005100975229 A CN B2005100975229A CN 200510097522 A CN200510097522 A CN 200510097522A CN 100526305 C CN100526305 C CN 100526305C
- Authority
- CN
- China
- Prior art keywords
- cell
- compound
- neural stem
- propolin
- neural
- 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.)
- Active
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 86
- 230000012010 growth Effects 0.000 title claims description 14
- 230000001737 promoting effect Effects 0.000 title description 3
- 210000001178 neural stem cell Anatomy 0.000 claims abstract description 71
- 230000001537 neural effect Effects 0.000 claims abstract description 22
- 239000000126 substance Substances 0.000 claims abstract description 9
- 239000000835 fiber Substances 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 57
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 claims description 27
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 claims description 24
- 102000013275 Somatomedins Human genes 0.000 claims description 24
- 108010025020 Nerve Growth Factor Proteins 0.000 claims description 18
- 102000015336 Nerve Growth Factor Human genes 0.000 claims description 10
- OMFXVFTZEKFJBZ-HJTSIMOOSA-N corticosterone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@H](CC4)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 OMFXVFTZEKFJBZ-HJTSIMOOSA-N 0.000 claims description 10
- 230000004069 differentiation Effects 0.000 claims description 10
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 claims description 10
- 229940053128 nerve growth factor Drugs 0.000 claims description 9
- 239000000654 additive Substances 0.000 claims description 5
- 230000000996 additive effect Effects 0.000 claims description 5
- 229960000890 hydrocortisone Drugs 0.000 claims description 5
- 239000003102 growth factor Substances 0.000 claims description 4
- CCNNANWAOVGRMQ-XLPSYMGOSA-N (2s)-2-[3,4-dihydroxy-2-[(z)-7-hydroxy-3,7-dimethyloct-2-enyl]phenyl]-5,7-dihydroxy-2,3-dihydrochromen-4-one Chemical compound CC(O)(C)CCCC(/C)=C\CC1=C(O)C(O)=CC=C1[C@H]1OC2=CC(O)=CC(O)=C2C(=O)C1 CCNNANWAOVGRMQ-XLPSYMGOSA-N 0.000 claims description 3
- WCDJIWAYGSJPBT-LDEBPVJWSA-N (2s)-2-[3,4-dihydroxy-5-[(e)-7-hydroxy-3,7-dimethyloct-2-enyl]phenyl]-5,7-dihydroxy-2,3-dihydrochromen-4-one Chemical group OC1=C(O)C(C\C=C(CCCC(C)(C)O)/C)=CC([C@H]2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 WCDJIWAYGSJPBT-LDEBPVJWSA-N 0.000 claims description 3
- QEXZVESBFWVISP-UHFFFAOYSA-N 3'-geranyl-4',5,7-trihydroxyflavanone Natural products C1=C(O)C(CC=C(C)CCC=C(C)C)=CC(C2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 QEXZVESBFWVISP-UHFFFAOYSA-N 0.000 claims description 3
- AMSGNHVRVKYLPO-UHFFFAOYSA-N Isonymphaeol B Natural products OC1=C(O)C(CC=C(C)CCC=C(C)C)=CC(C2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 AMSGNHVRVKYLPO-UHFFFAOYSA-N 0.000 claims description 3
- SZHDIKOQLFZADP-SANMLTNESA-N Nymphaeol C Natural products CC(C)=CCCC(C)=CCC1=C(O)C(O)=CC=C1[C@H]1OC2=CC(O)=C(CC=C(C)C)C(O)=C2C(=O)C1 SZHDIKOQLFZADP-SANMLTNESA-N 0.000 claims description 3
- AMSGNHVRVKYLPO-CEMXSPGASA-N isonymphaeol B Chemical group OC1=C(O)C(C/C=C(C)/CCC=C(C)C)=CC([C@H]2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 AMSGNHVRVKYLPO-CEMXSPGASA-N 0.000 claims description 3
- ILCMVLORKWIOOH-CEMXSPGASA-N nymphaeol B Chemical compound CC(C)=CCC\C(C)=C\CC1=C(O)C(O)=CC=C1[C@H]1OC2=CC(O)=CC(O)=C2C(=O)C1 ILCMVLORKWIOOH-CEMXSPGASA-N 0.000 claims description 3
- SZHDIKOQLFZADP-XBPZWBIKSA-N nymphaeol C Chemical compound CC(C)=CCC\C(C)=C\CC1=C(O)C(O)=CC=C1[C@H]1OC2=CC(O)=C(CC=C(C)C)C(O)=C2C(=O)C1 SZHDIKOQLFZADP-XBPZWBIKSA-N 0.000 claims description 3
- CCNNANWAOVGRMQ-NRFANRHFSA-N propolin A Natural products CC(O)(C)CCCC(C)=CCC1=C(O)C(O)=CC=C1[C@H]1OC2=CC(O)=CC(O)=C2C(=O)C1 CCNNANWAOVGRMQ-NRFANRHFSA-N 0.000 claims description 3
- WCDJIWAYGSJPBT-NRFANRHFSA-N propolin B Natural products OC1=C(O)C(CC=C(CCCC(C)(C)O)C)=CC([C@H]2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 WCDJIWAYGSJPBT-NRFANRHFSA-N 0.000 claims description 3
- ILCMVLORKWIOOH-QFIPXVFZSA-N propolin D Natural products CC(C)=CCCC(C)=CCC1=C(O)C(O)=CC=C1[C@H]1OC2=CC(O)=CC(O)=C2C(=O)C1 ILCMVLORKWIOOH-QFIPXVFZSA-N 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 claims description 2
- 230000000638 stimulation Effects 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims 1
- XCYSQFHYFNWYFP-CEMXSPGASA-N nymphaeol A Chemical compound C1([C@H]2OC3=CC(O)=C(C(=C3C(=O)C2)O)C/C=C(C)/CCC=C(C)C)=CC=C(O)C(O)=C1 XCYSQFHYFNWYFP-CEMXSPGASA-N 0.000 claims 1
- XCYSQFHYFNWYFP-QFIPXVFZSA-N propolin C Natural products C1([C@H]2OC3=CC(O)=C(C(=C3C(=O)C2)O)CC=C(C)CCC=C(C)C)=CC=C(O)C(O)=C1 XCYSQFHYFNWYFP-QFIPXVFZSA-N 0.000 claims 1
- 210000002569 neuron Anatomy 0.000 abstract description 45
- 210000000130 stem cell Anatomy 0.000 abstract description 11
- 230000004083 survival effect Effects 0.000 abstract description 11
- 238000012258 culturing Methods 0.000 abstract description 5
- 230000010261 cell growth Effects 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 210000003061 neural cell Anatomy 0.000 abstract 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 24
- 238000000034 method Methods 0.000 description 18
- 239000001963 growth medium Substances 0.000 description 16
- 210000003710 cerebral cortex Anatomy 0.000 description 14
- 230000008569 process Effects 0.000 description 14
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 12
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 12
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical compound CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 description 12
- 125000000217 alkyl group Chemical group 0.000 description 12
- 229910052799 carbon Inorganic materials 0.000 description 12
- 229910052739 hydrogen Inorganic materials 0.000 description 12
- 239000001257 hydrogen Substances 0.000 description 12
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 12
- 125000001844 prenyl group Chemical group [H]C([*])([H])C([H])=C(C([H])([H])[H])C([H])([H])[H] 0.000 description 11
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 10
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 9
- 210000005036 nerve Anatomy 0.000 description 9
- 102000007072 Nerve Growth Factors Human genes 0.000 description 8
- 206010020718 hyperplasia Diseases 0.000 description 8
- 239000003900 neurotrophic factor Substances 0.000 description 8
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 description 5
- 210000001218 blood-brain barrier Anatomy 0.000 description 5
- 230000024245 cell differentiation Effects 0.000 description 5
- 230000005284 excitation Effects 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 229920000656 polylysine Polymers 0.000 description 5
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 4
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 4
- 102000034615 Glial cell line-derived neurotrophic factor Human genes 0.000 description 4
- 108091010837 Glial cell line-derived neurotrophic factor Proteins 0.000 description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 4
- 210000001130 astrocyte Anatomy 0.000 description 4
- 210000003618 cortical neuron Anatomy 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 229930003935 flavonoid Natural products 0.000 description 4
- 235000017173 flavonoids Nutrition 0.000 description 4
- 125000002350 geranyl group Chemical group [H]C([*])([H])/C([H])=C(C([H])([H])[H])/C([H])([H])C([H])([H])C([H])=C(C([H])([H])[H])C([H])([H])[H] 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- -1 isoprene flavonoid compound Chemical class 0.000 description 4
- 230000031700 light absorption Effects 0.000 description 4
- 230000032405 negative regulation of neuron apoptotic process Effects 0.000 description 4
- 101000996663 Homo sapiens Neurotrophin-4 Proteins 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 102100033857 Neurotrophin-4 Human genes 0.000 description 3
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 3
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 3
- 230000008499 blood brain barrier function Effects 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000011712 cell development Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 210000002241 neurite Anatomy 0.000 description 3
- 210000004248 oligodendroglia Anatomy 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 206010003694 Atrophy Diseases 0.000 description 2
- 102000009024 Epidermal Growth Factor Human genes 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
- 238000000134 MTT assay Methods 0.000 description 2
- 231100000002 MTT assay Toxicity 0.000 description 2
- 108090000742 Neurotrophin 3 Proteins 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000037444 atrophy Effects 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 230000002490 cerebral effect Effects 0.000 description 2
- 230000003412 degenerative effect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000003125 immunofluorescent labeling Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 208000015122 neurodegenerative disease Diseases 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 229940056360 penicillin g Drugs 0.000 description 2
- 229930008679 prenylflavonoid Natural products 0.000 description 2
- 150000007951 prenylflavonoids Chemical group 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000004114 suspension culture Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010056677 Nerve degeneration Diseases 0.000 description 1
- 102000004230 Neurotrophin 3 Human genes 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 208000010513 Stupor Diseases 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 210000005056 cell body Anatomy 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 210000001671 embryonic stem cell Anatomy 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 230000008472 epithelial growth Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000008035 nerve activity Effects 0.000 description 1
- 230000000626 neurodegenerative effect Effects 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 229940032018 neurotrophin 3 Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 238000011476 stem cell transplantation Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a compound that promotes the neural cell growth and neural stem cell generation, and it is prenylflavanones with its chemical structure being shown in right formula. The compound can increase the survival rate of neural cell, especially in the culturing condition with low density; it can also promote the neural cell growth and make the neural fiber be thicker and longer and increase its branches. The compound can also promote the formation of neural stem cell, and the formed stem cell is identified as neuronal stem cells and it can induce it to neurons.
Description
Technical field
The present invention relates to a kind of by promoting the regeneration and the differentiation of neural stem cell, to suppress neurodegenerative compound.
Background technology
Known neural stem cell (neural stem cell, NSCs) be meant and carry out the self reparation, and can be under condition of in vitro culture, be divided into the cell of multiple different kenels, for example neuronal cell (Neurons), astroglia cell (Astrocytes) and oligodendrocyte (Oligodendrocyte) by the stimulation of some stimulating factors.Cell after these differentiation is for the growth of Mammals nervus centralis, and all playing the part of important role in the neural function performance of adult animals.Known neural stem cell can be separated to sophisticated process in its growth from multiple Mammals (for example mouse, rat, pig and the mankind's) central nervous system.Wherein, the neural stem cell in the human central nervous system is similar with its homologous rodent.
The neural stem cell of mammal (neural stem cell) is removed and is present in developmental neural system China and foreign countries, also is present in the sophisticated organ simultaneously.Though point out according to previous research report, can derive neural stem cell from embryonic stem cell at present, the regulatory mechanism of endogenous neural stem cell is but still understood limited.
For the treatment of neurodegenerative disorders, manage to save neurocyte in damaged condition, and stimulate the regeneration of this neurocyte simultaneously, be ideal therapeutic strategy comparatively.At present existing people attempts repairing damaged cells by using neural stem cells transplantation, and carries out the possibility of " self " by activating endogenous neural stem cell so that neurocyte to be provided.
Though at present scientist has had some preliminary understandings for neural stem cell, as if neural stem cell will being applied in the reparation of neurocyte, then also need a kind ofly can control its hyperplasia effectively or be divided into method with specific function cell.Show that according to former many parts of research reports neural stem cell can be bred having under the culture condition of somatomedin.These somatomedins are at present known comprise Basic Fibroblast Growth Factor (Basic Fibroblast Growth Factor, bFGF) and Urogastron (Epithelial Growth Factor, EGF) etc.Known at serum-free but contain in the substratum of aforementioned somatomedin, under the state of suspension culture, neural stem cell can be assembled and formed so-called " neural ball " (neurospheres).On the other hand, if these somatomedins are removed, change an amount of some micromolecular active compounds, other other somatomedin or neurotrophic factor (neurotrophic factors) except that Basic Fibroblast Growth Factor or Urogastron of replenishing into, neural stem cell then may irriate and is broken up, cell through breaking up is mainly based on astroglia cell (more than 90%), and only having small part to break up becomes neuronal cell (below 10%).
In addition, result of study in the past shows, scientist has been found that many neurotrophic factors at present, it includes glial cell line-derived neurotrophic factor (Glial-derived NeurotrophicFactor, GDNF), Brain Derived Neurotrophic Factor (Brain-derived Neurotrophic Factor, BDNF), nerve growth factor (nerve growth factor, NGF), neurotrophic factor 3 (neurotrophin-3, NT3), neurotrophic factor 4 (NT4), and platelet-derived growth factor (Platelet-derived growth factor, PDGF) etc.Have the activity that promotes the neurocyte survival though these neurotrophic factors have been proved, but it there is the restriction in many uses clinically.Therefore and be difficult for passing hemato encephalic barrier (Blood BrainBarrier BBB) arrives brain this is because when using these neurotrophic factors in the live body, and these neurotrophic factors are because have bigger molecular weight.
Therefore, if can find out some has than small molecular weight and can activate the compound of endogenous neural stem cell, in order to the hyperplasia that promotes neural stem cell and keep its characteristic, or even promote it to be divided into neuronal cell with function, this will help neural stem cells transplantation to host, so that effective prevention or the therapeutic strategy as the degenerative sacred disease to be provided, and provide preventive effect to some individualities with nerve degeneration factor.
Summary of the invention
For overcoming the shortcoming of known technology, the purpose of this invention is to provide that a kind of it has less molecular weight in order to promote the outgrowth compound of neurocyte, therefore facilitate penetration of hemato encephalic barrier and arrive brain, and then suppressed the death of neurocyte, and increase its survival rate.
Another object of the present invention provides a kind of in order to promote the compound of cell differentiation of nerve cord, is the neuronal cell with specific function to impel cell differentiation of nerve cord.
To achieve the object of the present invention, pointed a kind of in order to promote the outgrowth compound of neurocyte according to the present invention, it is Prenylflavonoid (prenylflavanones) compound that has as shown in the formula chemical structural formula shown in:
(formula one)
Wherein, R
1Be hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon or prenyl (isoprene); R
2, R
3, R
4, R
5, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon, prenyl (isoprene) or suc as formula two or formula three shown in geranyl (geranyl).
(formula two)
Known, neurocyte is at low density (160cells/mm
2Below) situation under when cultivating, neurocyte is easy to death, is difficult for cultivating.And according to the present invention pointed compound except that can even under low-density cultivation situation, also reducing the death of neurocyte significantly in order to promote the neuron survival rate.In addition, The compounds of this invention also can be in order to promote the growth of neurocyte, for example can make spinous process chap and elongated.In addition, The compounds of this invention can be in order to promoting the formation of neural stem cell, and can induce it to be divided into neuronal cell.
Description of drawings
Fig. 1 adds the mode of appearance photographic view after the culture medium culturing that contains The compounds of this invention and known somatomedin for neural stem cell: (A) compd A; (B) compd B; (C) Compound C; (D) Compound D; (E) compd E; (F) compound F 17-hydroxy-corticosterone; (G) compound G; (H) compound H; (I) blank blank group; (J) Urogastron; (K) nerve growth factor.
Fig. 2 is for adding the interpretation of result figure (* represent p＜0.0005) of The compounds of this invention to the influence of cerebral cortex neurons cell growth in substratum.
(* represents p＜0.005 to the interpretation of result figure of the influence when Fig. 3 cultivates with different cell densities the cerebral cortex neurons cell for The compounds of this invention; * represents p＜0.05): (A) cell density 300cells/mm
2(B) cell density 150cells/mm
2
Fig. 4 is The compounds of this invention influences gained to the neurite lengths of neurocyte a photographic view: (A) blank group; (B) compd A; (C) compd B; (D) Compound D; (E) compound G.
Fig. 5 is for being induced the photographic view of the situation gained after the differentiation with the microscopic examination neural stem cell, wherein left bank is the photomap fluorescence excitation under, and right row is the cell kenel under the visible light: (A) elder generation then cultivates with compd A with the compd A cultivation again; (B) cultivate with compd A earlier, afterwards with the culture medium culturing of not adding any somatomedin; (C) cultivate with Urogastron earlier, cultivate with Urogastron more afterwards; (D) cultivate with Urogastron earlier, afterwards with the culture medium culturing of not adding any somatomedin.
Fig. 6 is for being induced the photographic view of the situation gained after the differentiation with the microscopic examination neural stem cell, wherein left bank is the photomap fluorescence excitation under, and arrange on the right side is cell kenel under the visible light: (A) Compound D+Urogastron; (B) Compound D; (C) Urogastron; (D) do not add any somatomedin.
Fig. 7 is the photographic view of the mode of appearance of neural stem cell after cultivating 30 days: (A) blank group; (B) add compd A; (C) add compd A, the photomap under fluorescence excitation.
The present invention will be by being described further with reference to following embodiment, and embodiment described here does not limit the disclosed content in front of the present invention.Those skilled in the art can do more a little improvement and modify, but it does not still depart from the scope of the present invention.
Embodiment
Pointed a kind of in order to promote the outgrowth compound of neurocyte according to the present invention, it is the isoprene flavonoid compound that has as shown in the formula chemical structural formula shown in:
Wherein, R
1Be hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon or prenyl (isoprene); R
2, R
3, R
4, R
5, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon, prenyl (isoprene) or suc as formula two or formula three shown in geranyl (geranyl).
(formula two)
The aforesaid isoprene flavonoid compound of the present invention can be synthetic by known organic chemistry synthetic technology, also can obtain after modifying part functional group with known chemical modification technology by isolate the compound with approximate chemical structure in natural goods again.
As the aforementioned isoprene flavonoid compound specific embodiment of the present invention, it has following chemical structural formula:
Wherein, R
1, R
2, R
3, R
5, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl or the prenyl of 1～3 carbon.
Or
(formula five)
Wherein, R
1, R
2, R
3, R
5, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl or the prenyl of 1～3 carbon.
Or
Wherein, R
1, R
2, R
3, R
4, R
5With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon,
Wherein, R
1, R
2, R
3, R
4, R
5With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl or the prenyl of 1～3 carbon.
Or
(formula eight)
Wherein, R
1, R
2, R
3, R
4, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon, or prenyl.
Or
Wherein, R1, R2, R3, R4, R6 and R7 be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl of 1～3 carbon, or prenyl.
Or
(formula ten)
Wherein, R
1, R
3, R
4, R
5, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl or the prenyl of 1 ~ 3 carbon.
Or
(formula 11)
Wherein, R
1, R
3, R
4, R
5, R
6With R
7Be respectively hydrogen (H), hydroxyl (OH), the straight or branched alkyl or the prenyl of 1 ~ 3 carbon.
Aforementioned in order to promote further embodiment of the outgrowth compound of neurocyte as the present invention, it has following chemical structural formula:
Compd A (Propolin A)
(formula 12)
Compd B (Propolin B)
(formula 13)
(formula 14)
Compound D (Propolin D)
(formula 15)
(formula 16)
Compound F 17-hydroxy-corticosterone (Propolin F)
Compound G (Propolin G)
Compound H (Propolin H)
(formula 19)
Pointed compound has following characteristics according to the present invention:
(1) The compounds of this invention can be at low density (160cells/mm
2Below) under the culture condition of neurocyte, promote the growth of neurocyte (for example brain cortex neurocyte) significantly, to improve the neuron survival rate.
(2) in the influence to nerve growth and growth, The compounds of this invention can obtain thicker, the longer more spinous process of multiple-limb that reaches than handling with Basic Fibroblast Growth Factor or Urogastron significantly.
(3) aspect the generation that promotes neural stem cell, handle with The compounds of this invention, significantly than handling with Basic Fibroblast Growth Factor or Urogastron, more can promote the generation of neural stem cell, and can keep this spherical characteristics and under the situation of vitro culture, survive at least 30 days, and these neural stem cell can be induced to differentiate into neuronal cell, astroglia cell and oligodendroglia.
(4) The compounds of this invention can be in order to inducing the neural stem cell (neural stem cells) of generation based on neuronal stem cell (neuronal stemcells), and impel these stem cells further to be divided into neuronal cell (neurons).In addition, The compounds of this invention is different with known somatomedin (growth factors), therefore can be on using by the common Basic Fibroblast Growth Factor (basic fibroblast growth factor) that uses, with the hyperplasia of carrying out neural stem cell effectively neural stem cell is quantized, promptly can obtain neuronal stem cell in large quantities with this, can be applicable to treat the cell therapy of degenerative sacred disease future.
Because The compounds of this invention has aforesaid function and characteristics, therefore The compounds of this invention can be used as in the additive of vitro culture neural stem cell, with the employed macro-molecular protein of aforementioned known techniques (for example, Urogastron, the growth of alkaline fiber parent cell, glial cell line-derived neurotrophic factor, Brain Derived Neurotrophic Factor, nerve growth factor, neurotrophic factor 3, neurotrophic factor 4, and platelet-derived growth factor etc.) difference, isoprene flavonoid compound in the The compounds of this invention is the less small-molecule substance of molecular weight, and it can keep its characteristic (changed a subculture and get final product in about 7～9 days) more for a long time in nutrient solution.In addition, but the The compounds of this invention induced nerve stem cells is divided into neuronal cell significantly.In addition, those skilled in the art also can learn after reading specification sheets of the present invention, also can further comprise a growth factor in the The compounds of this invention, is used for culture of neural stem cells neural, to impel its hyperplasia.The example that aforesaid somatomedin can be enumerated in the present invention comprises Urogastron, Basic Fibroblast Growth Factor and nerve growth factor, but is not limited in this.This research for some neuroscience aspects is the cell cultures additive of a better and emerging experiment with serum-free.
In addition, the known small molecules material is more easily by hemato encephalic barrier (Blood Brain Barrier, BBB) arrive brain cell, and present U.S. food and FAD (Food and DrugAdministration, FDA) just actively carry out approval about human stem cell transplantation treatment mode, especially about the treatment of neurodegenerative disorders aspect, and the Prenylflavonoid that is comprised in the The compounds of this invention is micromolecular material, and it can promote to have the formation of functional neuronal cell in the substratum of serum-free, and can under the situation of low cell density, promote the existence of neurocyte, therefore these characteristics that The compounds of this invention had, treatment for auxiliary aforementioned human stem cell is transplanted can provide very big help.
In addition, those skilled in the art also can recognize that The compounds of this invention can further become a pharmaceutical compound by known technological development by reading aforesaid explanation.
Embodiment 1
The growth medium of preparation culture of neural stem cells neural, it is at B-27 serum-free neuronal cell cultures base (B-27 supplemented neurobasal medium, the L-glutamic acid of interpolation penicillin G (penicillin G), Vetstrep (streptomycin sulphate) and 0.5mM Gibco) (L-glutamine, Gibco).
Under narcosis, from female mouse (Wistar) abdominal cavity of conceived 16 days, take out the unborn tire mouse in foetal sac.Take out the cerebral tissue of tire mouse, act on 1 minute down at 25 ℃ with 0.1% trypsin solution (trypsinsolution).After phosphate buffer soln (PBS solution) washing 3 times, mix up and down that with known mechanical system its cell is broken up.Afterwards, with the aforementioned solution that contains tire mouse cerebral tissue cell by 70 μ m nylon cell filter screens (nylon cell strainer, Falcon), so that neural stem cell is discharged in the solution.With this solution centrifugal 10 minutes,, can obtain to comprise the suspension of neural stem cell cell mass so afterwards again with the growth medium displacement of aforementioned culture of neural stem cells neural with the rotating speed of 1000rpm.
With 150, the cell concentration of 000cells is positioned on the culture dish (Petri dish) with the suspension of aforementioned neural stem cell, and with it at 37 ℃, 5%CO
2, and cultivate under the environment of relative humidity 95%, with culture of neural stem cells neural.The growth medium of culturing cell was changed 1 time in per three days, changed the nutrient solution of half at every turn.
When experimentizing, with aforementioned neural stem cell with 300cells/mm
2Cell concentration be positioned in the culture dish of the growth medium that is mounted with neural stem cell.Wherein, in the growth medium of blank group, further add Urogastron (5ng/mL), Basic Fibroblast Growth Factor (5ng/mL) or nerve growth factor (5ng/mL).Then add The compounds of this invention A～H in the growth medium of experimental group respectively.Then add in the growth medium of blank group 5 μ L dimethyl sulfoxide (DMSO) (Dimethyl Sulfoxide, DMSO).Afterwards, cultivate at above-mentioned culture condition, and after cultivating seven days with the size of microscopic examination suspension neural stem cell, outer kenel and survival rate.Gained the results are shown in Fig. 1.
The result shows that The compounds of this invention A～H, Urogastron, nerve growth factor can make neural stem cell be gathered into spherical (neural ball), and bigger than the formed neural ball of blank group.Can find out significantly that thus The compounds of this invention can reach the purpose of identical promotion neural stem cell growth with known somatomedin.Just, The compounds of this invention has promoted effect for the hyperplasia of neural stem cell globe.
Embodiment 2
With the neural stem cell of gained among the embodiment 1 with 300cells/mm
2Cell concentration cultivate that (poly-D-lysine is in 6 hole culture plates Sigma), at 37 ℃, 5% CO adsorbing 30 μ g/mL poly-lysines
2, and cultivate under the environment of relative humidity 95%.The cell that forms through the cultivation differentiation is referred to as cerebral cortex neurons cell (cortical neurons).
Aforesaid cerebral cortex neurons cell is incubated at respectively in the growth medium that contains 10 μ M compd As, compd B, Compound D, compd E, compound F 17-hydroxy-corticosterone, compound G and compound H, repeats three tests.In addition, the dimethyl sulfoxide (DMSO) that adds 5 μ L in the growth medium of control group.After cultivating five days, carry out cell counting (MTT assay) with the method for known living cell counting number, and carry out result's analysis at the 540nm light absorption value, the results are shown in Fig. 2.
Result by Fig. 2 gained shows that the light absorption value of experimental group is significantly than the height of blank group, and is wherein best with the effect of compd A, D and G again.This result shows that The compounds of this invention can increase the survival rate of cerebral cortex neurons cell really significantly.
Embodiment 3
With the neural stem cell of gained among the embodiment 1 respectively with different cell densities (150,300cells/mm
2) cell concentration cultivate that (poly-D-lysine is in 6 hole culture plates Sigma), at 37 ℃, 5% CO adsorbing 30 μ g/mL poly-lysines
2, and cultivate under the environment of relative humidity 95%.The cell that forms through the cultivation differentiation is referred to as cerebral cortex neurons cell (cortical neurons).
Aforementioned cerebral cortex neurons cell is incubated at respectively in the growth medium that contains compd A, D and G, repeats three tests.In addition, the dimethyl sulfoxide (DMSO) of getting 5 μ L adds in the growth medium, in contrast group.Cultivate after five days, carry out cell counting (MTT assay), and carry out result's analysis, the results are shown in Fig. 3 in the 540nm light absorption value with the method for known living cell counting number.Known cerebral cortex neurons cell is with 640cells/mm
2Cell density when being incubated in the growth medium, the survival rate of cell can be higher than 90%, and with 160cells/mm
2Cell density when cultivating, the survival rate of cell can reduce to about 50%.Therefore cell density is lower than 160cells/mm when cultivating
2The time, if do not add any somatomedin in good time, then the mortality ratio of cell will increase significantly.This is that meeting is too far away owing to cell count makes cell and intercellular distance be separated by very little, and causes cell and iuntercellular to be difficult for carrying out the transmission of somatomedin because in this case.
Result by Fig. 3 gained shows, be added with in the experimental group of The compounds of this invention, no matter initial cultured cells density how, the light absorption value of its gained is all significantly than the height of control group, this result shows the interpolation of The compounds of this invention, can improve the neuron survival rate of cultivating under low cell density significantly.
Embodiment 4
With the neural stem cell of gained among the embodiment 1 with 38cells/mm
2Cell concentration be incubated at and adsorb 30 μ g/mL poly-lysines (poly-D-lysine is in 6 hole culture plates Sigma), at 37 ℃, 5% CO
2, and cultivate under the environment of relative humidity 95%.The cell that forms through the cultivation differentiation is referred to as cerebral cortex neurons cell (cortical neurons).
With aforementioned with the cerebral cortex neurons cell cultures in the substratum that contains The compounds of this invention A, B, D and G.In addition, get in the dimethyl sulfoxide (DMSO) adding growth medium of 5 μ L, to compare group.After cultivating five days, carry out cell observation and calculate the length of spinous process (neurite) with microscope, gained the results are shown in Fig. 4.
With reference to figure 4, do not add the blank group of any somatomedin fully, can be observed the atrophy of its neurone body (soma) at microscopically, the situation (Fig. 4 A) that neurocyte is dead fully is so the length of its measured spinous process is zero.Yet, experimental group is after adding compd A 2.5 μ M, compd B 5 μ M, Compound D 5 μ M and compound G5 μ M respectively, the neuronal cell that can be observed in the experimental group at microscopically continues growth, and spinous process outwards extends, and can be observed a neurone body and have at least the spinous process more than three to stretch out, and wherein one can be elongated and the tail end fork be a plurality of nerve synapses.Neurite lengths is analyzed, and behind the adding compd A, spinous process L1 length is that 236 μ m, spinous process L2 length are 161 μ m (Fig. 4 B); After adding compd B, spinous process L1 length is 235 μ m (Fig. 4 C); After adding compd B among the picture D, spinous process L1 length is 211 μ m (Fig. 4 D); After adding compd B among the picture E, spinous process L1 length is that 316 μ m, spinous process L2 length are 216 μ m (Fig. 4 E).
The result can learn that The compounds of this invention can be at low cell concn (38cells/mm thus
2) under, promote neuron survival, and impel its nerve process elongation, and growth is sophisticated neuronal cell.Embodiment 5
From the cell suspending liquid of embodiment 1 gained, take out 100 μ L, it is diluted to 1mL.Then, 30 μ g/mL poly-lysine (poly-D-lysine are being adsorbed in the supernatant liquor cultivation of drawing 250 μ L, Sigma) on the slide glass, comply with aforesaid culture condition and cultivate after 3 days, so that it is divided into cerebral cortex neurons cell (cortical neurons).Dye by known immunofluorescence staining at last.When carrying out immunofluorescence dyeing, use can be anti-as one with the anti-MAP2 of neuronal cell specificity bonded, and use that can to discern an anti-mouse immune globulin antibody that contains fluorescence matrix anti-as two.After neuronal cell is by an anti-identification, can make neuronal cell under the exciting of fluorescence, produce fluorescence.
When the beginning culture of neural stem cells neural, add compd A or Urogastron, then taking out neural stem cell is attached on the slide glass, then after cell all is attached on the slide, removing enchylema adds in the nutrient solution that contains compd A or Urogastron again, or do not add in the nutrient solution of any somatomedin, cultivate and use immunofluorescence dyeing observation of cell kenel after three days.
With reference to figure 5, left bank be under the fluorescence excitation photomap (white light for neuronal cell, the part of shade be dead cell or is non-neuronal cell among the figure, right row is cell kenel under the visible light, it can manifest all cells under this visual field.Therefore but actual observation is seen from the result of the 5th figure, when culture of neural stem cells neural at the beginning, add compd A, whether add compd A when no matter later on cytodifferentiation becomes the cerebral cortex neurons cell, the capital makes cell differentiation of nerve cord become neuronal cell (Fig. 5 A, B), and when the cell adding epidermal growth factor period of the day from 11 p.m. to 1 a.m, can make neurocyte hyperplasia (Fig. 5 C), what but it was longer is non-neuronal cell mostly, and even if Urogastron no longer is provided afterwards, what cell was longer also is that non-neuronal cell is just than lack (Fig. 5 D) that continues adding mostly.Though can learn that thus Urogastron can be in order to promote the hyperplasia of neurocyte, its survival rate is increased, but its outgrowth cell mostly be the precursor cell of non-neuronal cell greatly, but The compounds of this invention is except that the survival that can increase neural stem cell, also can further influence its differentiation, make its neuralward unit cell development.
Embodiment 6
Experimental implementation is with embodiment 6, but when the beginning culture of neural stem cells neural, change into and add Compound D and use Urogastron simultaneously, then take out stem cell and be attached on the slide glass, treat that then cell all is attached on the slide after, remove enchylema and add again and contain Compound D, or Urogastron, or add the nutrient solution that does not contain any somatomedin, or add Compound D and use Urogastron simultaneously, cultivate and use immunofluorescence dyeing observation of cell kenel after three days.
With reference to figure 6, left bank be under the fluorescence excitation photomap (white light for neuronal cell, the part of shade be dead cell or is non-neuronal cell among the figure, right row is cell kenel under the visible light, it can manifest all cells under this visual field.But therefore from the result of Fig. 6 actual observation to, if add Compound D and Urogastron during culture of neural stem cells neural at the beginning, can be observed the remarkable outgrowth situation of neural stem cell, and after differentiation neuralward unit cell development (Fig. 6 A).And after cytodifferentiation becomes the cerebral cortex neurons cell, only adding Urogastron, its cell can be divided into non-neuronal cell (Fig. 6 C), and contains Compound D, then can impel its cell differentiation of nerve cord to become neuronal cell (Fig. 6 B).In addition, after cytodifferentiation becomes the cerebral cortex neurons cell, no longer add any somatomedin, also can make cell differentiation of nerve cord become neuronal cell, but the quantity of its neuronal cell can be than lack (Fig. 6 D) that adds Compound D again.Hence one can see that, and Urogastron can promote the hyperplasia of neurocyte, if use The compounds of this invention simultaneously then can make more cell neuralward unit cell development.Experimental result can further be learnt thus, when neural stem cell has added The compounds of this invention when growing, can make neural stem cell neuralward unit stem cell (neuronal stem cells) direction grow.
Embodiment 7
Get by the neural stem cell of turning out with compd A among the embodiment 1 and carry out cultured continuously, per seven days replacing one subcultures.In addition, with dimethyl sulfoxide (DMSO) as the blank group.When being cultured to the 30 day, the enchylema of sucking-off 100 μ L neural stem cell is diluted to 1mL with it.Afterwards, draw 250 μ L supernatant liquors, it is incubated at adsorbs 30 μ g/mL poly-lysines (poly-D-lysine, on slide glass Sigma), with the outward appearance kenel of microscopic examination neural stem cell, gained the results are shown in Fig. 7.
With reference to figure 7, can see the cell that does not almost have survival in the control group (Fig. 7 A) that does not add any somatomedin fully by Fig. 7, and can't present normal neural stem cell and can be gathered into the globular characteristic, presented the state of atrophy during cell suspension culture.The neural stem cell that the adding compd A is cultivated then still can be kept it and be gathered into spherical characteristic (Fig. 7 B), and this cell continue cultivated use immunofluorescence staining to dye after three days as embodiment 5, can see still that at microscopically neural stem cell can normally be divided into neuronal cell, and continue to carry out the growth (Fig. 7 C) of spinous process.
Comprehensive the foregoing description as can be known, The compounds of this invention can be used for long-time culture of neural stem cells neural and can keep it being gathered into spherical characteristic, and still can make neural stem cell neuralward unit cytodifferentiation.On the other hand, The compounds of this invention can be when cultured continuously, can change in per seven days under the situation of a subculture and cultivate for a long time, and can culture of neural stem cells neural make its survival surpass more than 30 days.Yet known somatomedin promptly need be changed a subculture in general about 3～4 days, and this is because known somatomedin is all macro-molecular protein, therefore is easy to lose activity at normal temperatures, and the characteristic of micromolecular compound of the present invention is not subjected to the influence of this factor.
Claims (6)
1. the additive of the substratum of a culture of neural stem cells neural, it comprises the compound with chemical structural formula as follows:
Compd A is Propolin A
Compd B is Propolin B
Compound C is Propolin C
Compound D is Propolin D
Compound F 17-hydroxy-corticosterone is Propolin F
Compound G is Propolin G
Compound H is Propolin H
Or the combination of above-claimed cpd.
2. the described additive of claim 1, wherein this substratum further comprises a growth factor.
3. the described additive of claim 2, wherein this somatomedin is selected from Urogastron, the growth of alkaline fiber parent cell, and the group formed of nerve growth factor.
4. have the compound of chemical structural formula as follows or the application in its medicine that is combined in the differentiation of preparation stimulation of neural stem cells:
Compd A is Propolin A
Compd B is Propolin B
Compound D is Propolin D
Compound F 17-hydroxy-corticosterone is Propolin F
Compound G is Propolin G
Compound H is Propolin H
5. the described application of claim 4, wherein, described application comprises common use somatomedin.
6. the described application of claim 5, wherein, this somatomedin is selected from Urogastron, the growth of alkaline fiber parent cell, reaches the group that nerve growth factor is formed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100975229A CN100526305C (en) | 2005-12-28 | 2005-12-28 | Compound for promoting neurocyte growth |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100975229A CN100526305C (en) | 2005-12-28 | 2005-12-28 | Compound for promoting neurocyte growth |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1990480A CN1990480A (en) | 2007-07-04 |
| CN100526305C true CN100526305C (en) | 2009-08-12 |
Family
ID=38213043
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100975229A Active CN100526305C (en) | 2005-12-28 | 2005-12-28 | Compound for promoting neurocyte growth |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100526305C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7585892B2 (en) | 2006-03-30 | 2009-09-08 | Naturewise Biotech & Medicals Corporation | Compound for promoting the growth of neural cells |
| JP5421524B2 (en) * | 2007-09-14 | 2014-02-19 | ナチュレワイズ バイオテック＆メディカル コーポレーション | Compounds for inhibiting histone deacetylase |
-
2005
- 2005-12-28 CN CNB2005100975229A patent/CN100526305C/en active Active
Non-Patent Citations (8)
| Title |
|---|
| "Cytotoxic flavanones of Schizolaena hystrix fromtheMadagascar rainforest",. Brian T. et al.J. Nat. Prod.,Vol.68 . 2005 |
| "Cytotoxic flavanones of Schizolaena hystrix fromtheMadagascar rainforest",. Brian T. et al.J. Nat. Prod.,Vol.68. 2005 * |
| "Geographic and genetic variation in the leaf surface resincomponents of Mimulus aurantiacus from southern California",. J. D. Hare.Biochemical Systematics and Ecology,Vol.30 . 2002 |
| "Geographic and genetic variation in the leaf surface resincomponents of Mimulus aurantiacus from southern California",. J. D. Hare.Biochemical Systematics and Ecology,Vol.30. 2002 * |
| A new prenylated flavonoid from propolis collected inOkinawa, Japan. Shigenori Kumazawa, et al.Biosci. Biotechnol. Biochem.,,Vol.68 No.1. 2004 |
| A new prenylated flavonoid from propolis collected inOkinawa, Japan. Shigenori Kumazawa, et al.Biosci. Biotechnol. Biochem.,Vol.68 No.1. 2004 * |
| Allelopathic potential of macaranga tanarius (L.) Muell. -arg.. Mei-Huims Tseng, et al.Journal of Chemical Ecology,Vol.29 No.5. 2003 |
| Propolin C from propolis induces apoptosis through activatingcaspases, Bid and cytochrome c release in human melanomacells. Chia-Nan Chen, et al.Biochemical Pharmacology,,Vol.67 . 2004 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1990480A (en) | 2007-07-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kempermann et al. | Neurogenesis in the adult hippocampus | |
| CN104726406B (en) | It is a kind of to induce the method that dental pulp Derived from Mesenchymal Stem Cells is nerve cell | |
| CN103484432B (en) | A kind of induced nerve stem/ancestor cell differentiates is the inducing culture of oligodendrocyte precursor cells and induction method thereof and purposes | |
| CN114317269B (en) | Multi-organ chip and application thereof in drug evaluation | |
| CN103789265A (en) | Method for efficiently separating and culturing hippocampal neurons | |
| CN100526305C (en) | Compound for promoting neurocyte growth | |
| CN106754718A (en) | A kind of human embryo stem cell slewing is broken up to the method for retinal pigment epithelium | |
| CN102703387A (en) | Astrocyte separating and cultivating method | |
| Zhang et al. | Functional and histological improvement of the injured spinal cord following transplantation of Schwann cells transfected with NRG1 gene | |
| CN103789267A (en) | Improved primary culture method for hippocampal neurons | |
| CN103805565B (en) | Hippocampal neuron is separated and primary culture method and reagent | |
| Wang et al. | A new method of isolating spinal motor neurons from fetal mouse | |
| CN100344756C (en) | A method for in vitro derivativing nerve stem cell directional differentiation to cholinergic neuron | |
| JP4625419B2 (en) | Nerve cell growth and neural stem cell generation promoting compound | |
| CN103305458B (en) | Method for inducing cholinergic neurons from pluripotent stem cells | |
| CN108070558A (en) | A kind of preparation method of clinical grade neural stem cell | |
| EP1834952B1 (en) | Compound for promoting the growth of neural cells | |
| Kang et al. | Effectiveness of muscle basal lamina carrying neural stem cells and olfactory ensheathing cells in spinal cord repair | |
| US7585892B2 (en) | Compound for promoting the growth of neural cells | |
| CN109456937A (en) | A kind of culture medium preparing Endometrial stem cell and preparation method | |
| CN104845908B (en) | A kind of richness strontium hair weeds cells and its cultural method | |
| AU2006201584C1 (en) | Compound for promoting the growth of neural cells | |
| CN101824399B (en) | Method for inducing and differentiating into dopaminergic neuron from human amniotic sepithelial cell and application for separating obtained dopaminergic neuron | |
| CN110938599B (en) | Culture method of neural stem cells and application thereof | |
| CN116751747B (en) | Serum-free induction medium and induction method for promoting mesenchymal stem cells to differentiate into neurons |
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 | ||
| C35 | Partial or whole invalidation of patent or utility model | ||
| IP01 | Partial invalidation of patent right |
Commission number: 4W02899 Conclusion of examination: Claim No. 200510097522.9 of the patent right for invention No. first shall be invalid and the patent right shall be maintained on the basis of claim 2-6. Decision date of declaring invalidation: 20100702 Decision number of declaring invalidation: 15073 Denomination of invention: Compound promoting neurocyte growth Granted publication date: 20090812 Patentee: Naturewise Biotech & Medicals Corp. |