CN115006534B - 钾离子通道Kir4.1抑制剂治疗抑郁症的用途和药物组合物 - Google Patents
钾离子通道Kir4.1抑制剂治疗抑郁症的用途和药物组合物 Download PDFInfo
- Publication number
- CN115006534B CN115006534B CN202210060456.1A CN202210060456A CN115006534B CN 115006534 B CN115006534 B CN 115006534B CN 202210060456 A CN202210060456 A CN 202210060456A CN 115006534 B CN115006534 B CN 115006534B
- Authority
- CN
- China
- Prior art keywords
- depression
- seq
- sequence
- inhibitor
- precursor
- 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
- 108010074781 Kcnj10 (channel) Proteins 0.000 title claims abstract description 195
- 239000003112 inhibitor Substances 0.000 title claims abstract description 38
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 17
- 102100021176 ATP-sensitive inward rectifier potassium channel 10 Human genes 0.000 claims abstract description 162
- 239000003814 drug Substances 0.000 claims abstract description 26
- 238000011282 treatment Methods 0.000 claims abstract description 17
- 102000004257 Potassium Channel Human genes 0.000 claims abstract description 11
- 108020001213 potassium channel Proteins 0.000 claims abstract description 11
- 208000020401 Depressive disease Diseases 0.000 claims abstract description 9
- 230000014509 gene expression Effects 0.000 claims description 39
- 230000000694 effects Effects 0.000 claims description 38
- 230000002452 interceptive effect Effects 0.000 claims description 25
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 22
- 239000004055 small Interfering RNA Substances 0.000 claims description 19
- 108091027967 Small hairpin RNA Proteins 0.000 claims description 18
- 239000002243 precursor Substances 0.000 claims description 17
- 239000012634 fragment Substances 0.000 claims description 14
- 230000035772 mutation Effects 0.000 claims description 11
- 230000002829 reductive effect Effects 0.000 claims description 11
- 108020004999 messenger RNA Proteins 0.000 claims description 10
- 102100039280 Glycogenin-1 Human genes 0.000 claims description 7
- 150000007523 nucleic acids Chemical class 0.000 claims description 7
- 210000004492 nuclear pore Anatomy 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 108020004707 nucleic acids Proteins 0.000 claims description 5
- 102000039446 nucleic acids Human genes 0.000 claims description 5
- 108091026890 Coding region Proteins 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 34
- 238000012360 testing method Methods 0.000 abstract description 22
- 239000000126 substance Substances 0.000 abstract description 14
- 238000010171 animal model Methods 0.000 abstract description 12
- 238000012216 screening Methods 0.000 abstract description 4
- 241000700159 Rattus Species 0.000 description 60
- 210000002569 neuron Anatomy 0.000 description 58
- 210000001130 astrocyte Anatomy 0.000 description 48
- 210000004027 cell Anatomy 0.000 description 28
- 101150069149 LHB gene Proteins 0.000 description 24
- 241000700605 Viruses Species 0.000 description 21
- 230000006870 function Effects 0.000 description 21
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 20
- 238000002474 experimental method Methods 0.000 description 20
- 230000002401 inhibitory effect Effects 0.000 description 17
- 108090000623 proteins and genes Proteins 0.000 description 17
- 241000699670 Mus sp. Species 0.000 description 16
- 238000002347 injection Methods 0.000 description 16
- 239000007924 injection Substances 0.000 description 16
- 239000013612 plasmid Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 241000702423 Adeno-associated virus - 2 Species 0.000 description 15
- 230000001537 neural effect Effects 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 210000001519 tissue Anatomy 0.000 description 14
- 241001465754 Metazoa Species 0.000 description 13
- 210000004556 brain Anatomy 0.000 description 13
- 201000010099 disease Diseases 0.000 description 13
- 239000002158 endotoxin Substances 0.000 description 13
- 229920006008 lipopolysaccharide Polymers 0.000 description 13
- 230000036390 resting membrane potential Effects 0.000 description 13
- 239000003795 chemical substances by application Substances 0.000 description 12
- 230000002018 overexpression Effects 0.000 description 12
- 229940079593 drug Drugs 0.000 description 11
- 229910001414 potassium ion Inorganic materials 0.000 description 11
- UEQUQVLFIPOEMF-UHFFFAOYSA-N Mianserin Chemical compound C1C2=CC=CC=C2N2CCN(C)CC2C2=CC=CC=C21 UEQUQVLFIPOEMF-UHFFFAOYSA-N 0.000 description 10
- 230000000994 depressogenic effect Effects 0.000 description 10
- 238000010304 firing Methods 0.000 description 10
- 239000012528 membrane Substances 0.000 description 10
- 229960003955 mianserin Drugs 0.000 description 10
- 229960002073 sertraline Drugs 0.000 description 10
- VGKDLMBJGBXTGI-SJCJKPOMSA-N sertraline Chemical compound C1([C@@H]2CC[C@@H](C3=CC=CC=C32)NC)=CC=C(Cl)C(Cl)=C1 VGKDLMBJGBXTGI-SJCJKPOMSA-N 0.000 description 10
- 230000009182 swimming Effects 0.000 description 10
- 229960004038 fluvoxamine Drugs 0.000 description 9
- CJOFXWAVKWHTFT-XSFVSMFZSA-N fluvoxamine Chemical compound COCCCC\C(=N/OCCN)C1=CC=C(C(F)(F)F)C=C1 CJOFXWAVKWHTFT-XSFVSMFZSA-N 0.000 description 9
- 230000004927 fusion Effects 0.000 description 9
- 230000001105 regulatory effect Effects 0.000 description 9
- RTHCYVBBDHJXIQ-MRXNPFEDSA-N (R)-fluoxetine Chemical compound O([C@H](CCNC)C=1C=CC=CC=1)C1=CC=C(C(F)(F)F)C=C1 RTHCYVBBDHJXIQ-MRXNPFEDSA-N 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 8
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 8
- 239000000935 antidepressant agent Substances 0.000 description 8
- 230000006399 behavior Effects 0.000 description 8
- QWCRAEMEVRGPNT-UHFFFAOYSA-N buspirone Chemical compound C1C(=O)N(CCCCN2CCN(CC2)C=2N=CC=CN=2)C(=O)CC21CCCC2 QWCRAEMEVRGPNT-UHFFFAOYSA-N 0.000 description 8
- 229960002495 buspirone Drugs 0.000 description 8
- 230000002222 downregulating effect Effects 0.000 description 8
- 229960002464 fluoxetine Drugs 0.000 description 8
- 229910052700 potassium Inorganic materials 0.000 description 8
- 239000011591 potassium Substances 0.000 description 8
- 208000024891 symptom Diseases 0.000 description 8
- 238000001262 western blot Methods 0.000 description 8
- PHVGLTMQBUFIQQ-UHFFFAOYSA-N Nortryptiline Chemical compound C1CC2=CC=CC=C2C(=CCCNC)C2=CC=CC=C21 PHVGLTMQBUFIQQ-UHFFFAOYSA-N 0.000 description 7
- 229940005513 antidepressants Drugs 0.000 description 7
- 210000005056 cell body Anatomy 0.000 description 7
- 230000003001 depressive effect Effects 0.000 description 7
- 239000003550 marker Substances 0.000 description 7
- 229960001158 nortriptyline Drugs 0.000 description 7
- 241000702421 Dependoparvovirus Species 0.000 description 6
- 229940123445 Tricyclic antidepressant Drugs 0.000 description 6
- 230000000295 complement effect Effects 0.000 description 6
- 208000035475 disorder Diseases 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 210000004498 neuroglial cell Anatomy 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- 229940124834 selective serotonin reuptake inhibitor Drugs 0.000 description 6
- 239000003029 tricyclic antidepressant agent Substances 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 description 5
- 108020004459 Small interfering RNA Proteins 0.000 description 5
- 230000002159 abnormal effect Effects 0.000 description 5
- 230000003542 behavioural effect Effects 0.000 description 5
- 238000010276 construction Methods 0.000 description 5
- 230000000875 corresponding effect Effects 0.000 description 5
- 230000009368 gene silencing by RNA Effects 0.000 description 5
- 210000001320 hippocampus Anatomy 0.000 description 5
- 238000010166 immunofluorescence Methods 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 210000005036 nerve Anatomy 0.000 description 5
- 230000000638 stimulation Effects 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- 108091006146 Channels Proteins 0.000 description 4
- 102100039289 Glial fibrillary acidic protein Human genes 0.000 description 4
- 101710193519 Glial fibrillary acidic protein Proteins 0.000 description 4
- 101001092197 Homo sapiens RNA binding protein fox-1 homolog 3 Proteins 0.000 description 4
- 241000124008 Mammalia Species 0.000 description 4
- 102100035530 RNA binding protein fox-1 homolog 3 Human genes 0.000 description 4
- 101100075500 Rattus norvegicus Lhb gene Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 150000001413 amino acids Chemical group 0.000 description 4
- 230000001430 anti-depressive effect Effects 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 4
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 4
- 230000008045 co-localization Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 230000003828 downregulation Effects 0.000 description 4
- 210000005046 glial fibrillary acidic protein Anatomy 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 230000028161 membrane depolarization Effects 0.000 description 4
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 4
- 230000010412 perfusion Effects 0.000 description 4
- 230000001766 physiological effect Effects 0.000 description 4
- 239000012896 selective serotonin reuptake inhibitor Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000012549 training Methods 0.000 description 4
- 239000013603 viral vector Substances 0.000 description 4
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 3
- 206010054089 Depressive symptom Diseases 0.000 description 3
- 101000614696 Homo sapiens ATP-sensitive inward rectifier potassium channel 10 Proteins 0.000 description 3
- 108010021466 Mutant Proteins Proteins 0.000 description 3
- 241000700157 Rattus norvegicus Species 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 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 3
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 description 3
- 229910001626 barium chloride Inorganic materials 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000001276 controlling effect Effects 0.000 description 3
- 230000002596 correlated effect Effects 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 230000007831 electrophysiology Effects 0.000 description 3
- 238000002001 electrophysiology Methods 0.000 description 3
- 210000003722 extracellular fluid Anatomy 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 230000003137 locomotive effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 230000001256 tonic effect Effects 0.000 description 3
- 239000012103 Alexa Fluor 488 Substances 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 206010010356 Congenital anomaly Diseases 0.000 description 2
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 2
- 108060001084 Luciferase Proteins 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 102000008300 Mutant Proteins Human genes 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 108010033276 Peptide Fragments Proteins 0.000 description 2
- 102000007079 Peptide Fragments Human genes 0.000 description 2
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical class [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 2
- KJMOINFQVCCSDX-XKBZYTNZSA-N Ser-Gln-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KJMOINFQVCCSDX-XKBZYTNZSA-N 0.000 description 2
- 230000036982 action potential Effects 0.000 description 2
- 101150063416 add gene Proteins 0.000 description 2
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000005013 brain tissue Anatomy 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000007906 compression Methods 0.000 description 2
- 230000006835 compression Effects 0.000 description 2
- 230000001054 cortical effect Effects 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 2
- 230000000971 hippocampal effect Effects 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- 238000002513 implantation Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 229960002725 isoflurane Drugs 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 108010034529 leucyl-lysine Proteins 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 230000007762 localization of cell Effects 0.000 description 2
- 230000033001 locomotion Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 108091070501 miRNA Proteins 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000000944 nerve tissue Anatomy 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 229960001412 pentobarbital Drugs 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 208000020016 psychiatric disease Diseases 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- -1 small molecule compound Chemical class 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 206010001497 Agitation Diseases 0.000 description 1
- UCIYCBSJBQGDGM-LPEHRKFASA-N Ala-Arg-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(=O)O)N UCIYCBSJBQGDGM-LPEHRKFASA-N 0.000 description 1
- HGRBNYQIMKTUNT-XVYDVKMFSA-N Ala-Asn-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N HGRBNYQIMKTUNT-XVYDVKMFSA-N 0.000 description 1
- HMRWQTHUDVXMGH-GUBZILKMSA-N Ala-Glu-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN HMRWQTHUDVXMGH-GUBZILKMSA-N 0.000 description 1
- VNYMOTCMNHJGTG-JBDRJPRFSA-N Ala-Ile-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O VNYMOTCMNHJGTG-JBDRJPRFSA-N 0.000 description 1
- XHNLCGXYBXNRIS-BJDJZHNGSA-N Ala-Lys-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O XHNLCGXYBXNRIS-BJDJZHNGSA-N 0.000 description 1
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 1
- NZGRHTKZFSVPAN-BIIVOSGPSA-N Ala-Ser-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N NZGRHTKZFSVPAN-BIIVOSGPSA-N 0.000 description 1
- QKHWNPQNOHEFST-VZFHVOOUSA-N Ala-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C)N)O QKHWNPQNOHEFST-VZFHVOOUSA-N 0.000 description 1
- 239000012110 Alexa Fluor 594 Substances 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- HKRXJBBCQBAGIM-FXQIFTODSA-N Arg-Asp-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N HKRXJBBCQBAGIM-FXQIFTODSA-N 0.000 description 1
- MZRBYBIQTIKERR-GUBZILKMSA-N Arg-Glu-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MZRBYBIQTIKERR-GUBZILKMSA-N 0.000 description 1
- GXXWTNKNFFKTJB-NAKRPEOUSA-N Arg-Ile-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O GXXWTNKNFFKTJB-NAKRPEOUSA-N 0.000 description 1
- VIINVRPKMUZYOI-DCAQKATOSA-N Arg-Met-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O VIINVRPKMUZYOI-DCAQKATOSA-N 0.000 description 1
- AMIQZQAAYGYKOP-FXQIFTODSA-N Arg-Ser-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O AMIQZQAAYGYKOP-FXQIFTODSA-N 0.000 description 1
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 1
- OLISTMZJGQUOGS-GMOBBJLQSA-N Asn-Ile-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N OLISTMZJGQUOGS-GMOBBJLQSA-N 0.000 description 1
- XDGBFDYXZCMYEX-NUMRIWBASA-N Asp-Glu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(=O)O)N)O XDGBFDYXZCMYEX-NUMRIWBASA-N 0.000 description 1
- UZFHNLYQWMGUHU-DCAQKATOSA-N Asp-Lys-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UZFHNLYQWMGUHU-DCAQKATOSA-N 0.000 description 1
- HSGOFISJLFDMBJ-CIUDSAMLSA-N Asp-Met-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N HSGOFISJLFDMBJ-CIUDSAMLSA-N 0.000 description 1
- GWIJZUVQVDJHDI-AVGNSLFASA-N Asp-Phe-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O GWIJZUVQVDJHDI-AVGNSLFASA-N 0.000 description 1
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 208000036119 Frailty Diseases 0.000 description 1
- OACPJRQRAHMQEQ-NHCYSSNCSA-N Gln-Val-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O OACPJRQRAHMQEQ-NHCYSSNCSA-N 0.000 description 1
- HNAUFGBKJLTWQE-IFFSRLJSSA-N Gln-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCC(=O)N)N)O HNAUFGBKJLTWQE-IFFSRLJSSA-N 0.000 description 1
- NUSWUSKZRCGFEX-FXQIFTODSA-N Glu-Glu-Cys Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CS)C(O)=O NUSWUSKZRCGFEX-FXQIFTODSA-N 0.000 description 1
- OGNJZUXUTPQVBR-BQBZGAKWSA-N Glu-Gly-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O OGNJZUXUTPQVBR-BQBZGAKWSA-N 0.000 description 1
- ILWHFUZZCFYSKT-AVGNSLFASA-N Glu-Lys-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O ILWHFUZZCFYSKT-AVGNSLFASA-N 0.000 description 1
- IDEODOAVGCMUQV-GUBZILKMSA-N Glu-Ser-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IDEODOAVGCMUQV-GUBZILKMSA-N 0.000 description 1
- 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 1
- XTQFHTHIAKKCTM-YFKPBYRVSA-N Gly-Glu-Gly Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O XTQFHTHIAKKCTM-YFKPBYRVSA-N 0.000 description 1
- XPJBQTCXPJNIFE-ZETCQYMHSA-N Gly-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)CN XPJBQTCXPJNIFE-ZETCQYMHSA-N 0.000 description 1
- MHXKHKWHPNETGG-QWRGUYRKSA-N Gly-Lys-Leu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O MHXKHKWHPNETGG-QWRGUYRKSA-N 0.000 description 1
- CVFOYJJOZYYEPE-KBPBESRZSA-N Gly-Lys-Tyr Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CVFOYJJOZYYEPE-KBPBESRZSA-N 0.000 description 1
- FXLVSYVJDPCIHH-STQMWFEESA-N Gly-Phe-Arg Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FXLVSYVJDPCIHH-STQMWFEESA-N 0.000 description 1
- OOCFXNOVSLSHAB-IUCAKERBSA-N Gly-Pro-Pro Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 OOCFXNOVSLSHAB-IUCAKERBSA-N 0.000 description 1
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 1
- UVTSZKIATYSKIR-RYUDHWBXSA-N Gly-Tyr-Glu Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O UVTSZKIATYSKIR-RYUDHWBXSA-N 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- JWLWNCVBBSBCEM-NKIYYHGXSA-N His-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N)O JWLWNCVBBSBCEM-NKIYYHGXSA-N 0.000 description 1
- FSOXZQBMPBQKGJ-QSFUFRPTSA-N His-Ile-Ala Chemical compound [O-]C(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]([NH3+])CC1=CN=CN1 FSOXZQBMPBQKGJ-QSFUFRPTSA-N 0.000 description 1
- CCUSLCQWVMWTIS-IXOXFDKPSA-N His-Thr-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O CCUSLCQWVMWTIS-IXOXFDKPSA-N 0.000 description 1
- DRKZDEFADVYTLU-AVGNSLFASA-N His-Val-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O DRKZDEFADVYTLU-AVGNSLFASA-N 0.000 description 1
- OEQKGSPBDVKYOC-ZKWXMUAHSA-N Ile-Gly-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N OEQKGSPBDVKYOC-ZKWXMUAHSA-N 0.000 description 1
- UAQSZXGJGLHMNV-XEGUGMAKSA-N Ile-Gly-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N UAQSZXGJGLHMNV-XEGUGMAKSA-N 0.000 description 1
- PMMMQRVUMVURGJ-XUXIUFHCSA-N Ile-Leu-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(O)=O PMMMQRVUMVURGJ-XUXIUFHCSA-N 0.000 description 1
- RQQCJTLBSJMVCR-DSYPUSFNSA-N Ile-Leu-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N RQQCJTLBSJMVCR-DSYPUSFNSA-N 0.000 description 1
- YBKKLDBBPFIXBQ-MBLNEYKQSA-N Ile-Thr-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)O)N YBKKLDBBPFIXBQ-MBLNEYKQSA-N 0.000 description 1
- 108010009983 Inwardly Rectifying Potassium Channels Proteins 0.000 description 1
- 102000009855 Inwardly Rectifying Potassium Channels Human genes 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- RFUBXQQFJFGJFV-GUBZILKMSA-N Leu-Asn-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O RFUBXQQFJFGJFV-GUBZILKMSA-N 0.000 description 1
- CIVKXGPFXDIQBV-WDCWCFNPSA-N Leu-Gln-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CIVKXGPFXDIQBV-WDCWCFNPSA-N 0.000 description 1
- QVFGXCVIXXBFHO-AVGNSLFASA-N Leu-Glu-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O QVFGXCVIXXBFHO-AVGNSLFASA-N 0.000 description 1
- RZXLZBIUTDQHJQ-SRVKXCTJSA-N Leu-Lys-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O RZXLZBIUTDQHJQ-SRVKXCTJSA-N 0.000 description 1
- AUNMOHYWTAPQLA-XUXIUFHCSA-N Leu-Met-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O AUNMOHYWTAPQLA-XUXIUFHCSA-N 0.000 description 1
- VULJUQZPSOASBZ-SRVKXCTJSA-N Leu-Pro-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O VULJUQZPSOASBZ-SRVKXCTJSA-N 0.000 description 1
- IRMLZWSRWSGTOP-CIUDSAMLSA-N Leu-Ser-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O IRMLZWSRWSGTOP-CIUDSAMLSA-N 0.000 description 1
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-N 0.000 description 1
- SVBJIZVVYJYGLA-DCAQKATOSA-N Leu-Ser-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O SVBJIZVVYJYGLA-DCAQKATOSA-N 0.000 description 1
- ODRREERHVHMIPT-OEAJRASXSA-N Leu-Thr-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 ODRREERHVHMIPT-OEAJRASXSA-N 0.000 description 1
- SUYRAPCRSCCPAK-VFAJRCTISA-N Leu-Trp-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SUYRAPCRSCCPAK-VFAJRCTISA-N 0.000 description 1
- GDBQQVLCIARPGH-UHFFFAOYSA-N Leupeptin Natural products CC(C)CC(NC(C)=O)C(=O)NC(CC(C)C)C(=O)NC(C=O)CCCN=C(N)N GDBQQVLCIARPGH-UHFFFAOYSA-N 0.000 description 1
- AAORVPFVUIHEAB-YUMQZZPRSA-N Lys-Asp-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O AAORVPFVUIHEAB-YUMQZZPRSA-N 0.000 description 1
- IWWMPCPLFXFBAF-SRVKXCTJSA-N Lys-Asp-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O IWWMPCPLFXFBAF-SRVKXCTJSA-N 0.000 description 1
- PINHPJWGVBKQII-SRVKXCTJSA-N Lys-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCCN)N PINHPJWGVBKQII-SRVKXCTJSA-N 0.000 description 1
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 1
- ZJWIXBZTAAJERF-IHRRRGAJSA-N Lys-Lys-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CCCN=C(N)N ZJWIXBZTAAJERF-IHRRRGAJSA-N 0.000 description 1
- 235000000060 Malva neglecta Nutrition 0.000 description 1
- 240000000982 Malva neglecta Species 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- OLWAOWXIADGIJG-AVGNSLFASA-N Met-Arg-Lys Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(O)=O OLWAOWXIADGIJG-AVGNSLFASA-N 0.000 description 1
- GWADARYJIJDYRC-XGEHTFHBSA-N Met-Thr-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O GWADARYJIJDYRC-XGEHTFHBSA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 208000025966 Neurological disease Diseases 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- UEEVBGHEGJMDDV-AVGNSLFASA-N Phe-Asp-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UEEVBGHEGJMDDV-AVGNSLFASA-N 0.000 description 1
- MSHZERMPZKCODG-ACRUOGEOSA-N Phe-Leu-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MSHZERMPZKCODG-ACRUOGEOSA-N 0.000 description 1
- KNYPNEYICHHLQL-ACRUOGEOSA-N Phe-Leu-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 KNYPNEYICHHLQL-ACRUOGEOSA-N 0.000 description 1
- UNBFGVQVQGXXCK-KKUMJFAQSA-N Phe-Ser-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O UNBFGVQVQGXXCK-KKUMJFAQSA-N 0.000 description 1
- SHUFSZDAIPLZLF-BEAPCOKYSA-N Phe-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)O SHUFSZDAIPLZLF-BEAPCOKYSA-N 0.000 description 1
- FMLRRBDLBJLJIK-DCAQKATOSA-N Pro-Leu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FMLRRBDLBJLJIK-DCAQKATOSA-N 0.000 description 1
- YXHYJEPDKSYPSQ-AVGNSLFASA-N Pro-Leu-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 YXHYJEPDKSYPSQ-AVGNSLFASA-N 0.000 description 1
- WHNJMTHJGCEKGA-ULQDDVLXSA-N Pro-Phe-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WHNJMTHJGCEKGA-ULQDDVLXSA-N 0.000 description 1
- 108010057163 Ribonuclease III Proteins 0.000 description 1
- 102000003661 Ribonuclease III Human genes 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- MMAPOBOTRUVNKJ-ZLUOBGJFSA-N Ser-Asp-Ser Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CO)N)C(=O)O MMAPOBOTRUVNKJ-ZLUOBGJFSA-N 0.000 description 1
- ZIFYDQAFEMIZII-GUBZILKMSA-N Ser-Leu-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZIFYDQAFEMIZII-GUBZILKMSA-N 0.000 description 1
- VMLONWHIORGALA-SRVKXCTJSA-N Ser-Leu-Leu Chemical compound CC(C)C[C@@H](C([O-])=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]([NH3+])CO VMLONWHIORGALA-SRVKXCTJSA-N 0.000 description 1
- FKYWFUYPVKLJLP-DCAQKATOSA-N Ser-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FKYWFUYPVKLJLP-DCAQKATOSA-N 0.000 description 1
- SNXUIBACCONSOH-BWBBJGPYSA-N Ser-Thr-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CO)C(O)=O SNXUIBACCONSOH-BWBBJGPYSA-N 0.000 description 1
- 241001481798 Stochomys longicaudatus Species 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 206010042458 Suicidal ideation Diseases 0.000 description 1
- KGKWKSSSQGGYAU-SUSMZKCASA-N Thr-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KGKWKSSSQGGYAU-SUSMZKCASA-N 0.000 description 1
- SLUWOCTZVGMURC-BFHQHQDPSA-N Thr-Gly-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O SLUWOCTZVGMURC-BFHQHQDPSA-N 0.000 description 1
- PZSDPRBZINDEJV-HTUGSXCWSA-N Thr-Phe-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O PZSDPRBZINDEJV-HTUGSXCWSA-N 0.000 description 1
- HSQXHRIRJSFDOH-URLPEUOOSA-N Thr-Phe-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HSQXHRIRJSFDOH-URLPEUOOSA-N 0.000 description 1
- WNQJTLATMXYSEL-OEAJRASXSA-N Thr-Phe-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WNQJTLATMXYSEL-OEAJRASXSA-N 0.000 description 1
- DNCUODYZAMHLCV-XGEHTFHBSA-N Thr-Pro-Cys Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CS)C(=O)O)N)O DNCUODYZAMHLCV-XGEHTFHBSA-N 0.000 description 1
- HUPLKEHTTQBXSC-YJRXYDGGSA-N Thr-Ser-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HUPLKEHTTQBXSC-YJRXYDGGSA-N 0.000 description 1
- XGFYGMKZKFRGAI-RCWTZXSCSA-N Thr-Val-Arg Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N XGFYGMKZKFRGAI-RCWTZXSCSA-N 0.000 description 1
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- AKLNEFNQWLHIGY-QWRGUYRKSA-N Tyr-Gly-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N)O AKLNEFNQWLHIGY-QWRGUYRKSA-N 0.000 description 1
- QARCDOCCDOLJSF-HJPIBITLSA-N Tyr-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N QARCDOCCDOLJSF-HJPIBITLSA-N 0.000 description 1
- UEOOXDLMQZBPFR-ZKWXMUAHSA-N Val-Ala-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N UEOOXDLMQZBPFR-ZKWXMUAHSA-N 0.000 description 1
- FZSPNKUFROZBSG-ZKWXMUAHSA-N Val-Ala-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O FZSPNKUFROZBSG-ZKWXMUAHSA-N 0.000 description 1
- RUCNAYOMFXRIKJ-DCAQKATOSA-N Val-Ala-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN RUCNAYOMFXRIKJ-DCAQKATOSA-N 0.000 description 1
- DBOXBUDEAJVKRE-LSJOCFKGSA-N Val-Asn-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DBOXBUDEAJVKRE-LSJOCFKGSA-N 0.000 description 1
- DAVNYIUELQBTAP-XUXIUFHCSA-N Val-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N DAVNYIUELQBTAP-XUXIUFHCSA-N 0.000 description 1
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 1
- VVIZITNVZUAEMI-DLOVCJGASA-N Val-Val-Gln Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCC(N)=O VVIZITNVZUAEMI-DLOVCJGASA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 108010011559 alanylphenylalanine Proteins 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 206010003549 asthenia Diseases 0.000 description 1
- 210000003050 axon Anatomy 0.000 description 1
- 238000009227 behaviour therapy Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 1
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010045126 glycyl-tyrosyl-glycine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000002102 hyperpolarization Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- GDBQQVLCIARPGH-ULQDDVLXSA-N leupeptin Chemical compound CC(C)C[C@H](NC(C)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C=O)CCCN=C(N)N GDBQQVLCIARPGH-ULQDDVLXSA-N 0.000 description 1
- 108010052968 leupeptin Proteins 0.000 description 1
- 210000003715 limbic system Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 108010009298 lysylglutamic acid Proteins 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000036651 mood Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 230000008062 neuronal firing Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000001584 occupational therapy Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000007310 pathophysiology Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 108010091212 pepstatin Proteins 0.000 description 1
- FAXGPCHRFPCXOO-LXTPJMTPSA-N pepstatin A Chemical compound OC(=O)C[C@H](O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)C[C@H](O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)CC(C)C FAXGPCHRFPCXOO-LXTPJMTPSA-N 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 238000000554 physical therapy Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 210000004129 prosencephalon Anatomy 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 201000000980 schizophrenia Diseases 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000012453 sprague-dawley rat model Methods 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 238000011200 topical administration Methods 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- BPICBUSOMSTKRF-UHFFFAOYSA-N xylazine Chemical compound CC1=CC=CC(C)=C1NC1=NCCCS1 BPICBUSOMSTKRF-UHFFFAOYSA-N 0.000 description 1
- 229960001600 xylazine Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1138—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against receptors or cell surface proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- 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
- A61K31/13—Amines
- A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
-
- 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
- A61K31/13—Amines
- A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
- A61K31/138—Aryloxyalkylamines, e.g. propranolol, tamoxifen, phenoxybenzamine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4468—Non condensed piperidines, e.g. piperocaine having a nitrogen directly attached in position 4, e.g. clebopride, fentanyl
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5082—Supracellular entities, e.g. tissue, organisms
- G01N33/5088—Supracellular entities, e.g. tissue, organisms of vertebrates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
- G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/50—Physical structure
- C12N2310/53—Physical structure partially self-complementary or closed
- C12N2310/531—Stem-loop; Hairpin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/30—Psychoses; Psychiatry
- G01N2800/304—Mood disorders, e.g. bipolar, depression
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Urology & Nephrology (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Analytical Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Psychiatry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pain & Pain Management (AREA)
Abstract
本发明提供了钾离子通道蛋白Kir4.1抑制剂在制备治疗抑郁症的药物中的用途。本发明还提供了治疗抑郁症的药物组合物,其中包含Kir4.1抑制剂。本发明还提供了筛选治疗抑郁症的潜在物质的方法,包括给抑郁症动物模型施用待筛选的测试物的步骤,其中所述抑郁症动物模型的外侧缰核中Kir4.1是高表达的。
Description
本申请为2018年5月8日提交的、申请号为201810432839.0、发明名称为“钾离子通道抑制剂治疗抑郁症的用途和药物组合物”的分案申请,其全部内容通过引用结合在本申请中。
本申请要求2017年5月9日提交的、申请号为201710322245.X、发明名称为“Kir4.1抑制剂治疗抑郁症的用途和药物组合物”的中国专利申请的优先权,其全部内容通过引用结合在本申请中。
技术领域
本发明涉及疾病治疗和药物领域。具体的,本发明涉及抑郁症的治疗和用于治疗抑郁症的药物组合物和其制备方法。
背景技术
抑郁症是一种慢性精神障碍性疾病,以显著而持久的心境低落,动力缺失,行为绝望以及快感缺失为主要临床特征,严重的可表现出自杀倾向。
外侧缰核(LHb)近年来被认为是研究抑郁症病理生理学的关键脑区,在众多抑郁症的动物模型以及抑郁症病人中都发现了外侧缰核活性的显著升高。
神经元的异常活动主要归因于突触传递异常、自身生理特性变化以及神经元内环境的改变。星形胶质细胞参与调节神经细胞的自身活性,递质释放,而且在一系列包括精神分裂、癫痫、老年痴呆、抑郁症等在内的疾病中扮演重要角色(Hamilton et al.,Frontiersin neuroenergetics 2,2010)。对抑郁症死者大脑的解剖学研究表明,其前脑边缘系统中,胶质细胞的数目,形态及功能均发生显著变化(Cotter et al.,Archives of generalpsychiatry 58,545-553,2001;Coyle et al.,Archives of general psychiatry 57,90-93,2000;Rajkowska et al.,CNS&neurological disorders drug targets 6,219-233,2007)。利用化学手段诱导星形胶质细胞凋亡足以引起抑郁症状(Banasr et a l.,Biological psychiatry 64,863-870,2008)。抗抑郁药物能直接作用于星形胶质细胞,显著影响其形态及功能。该现象被认为可能是抗抑郁药物在中枢神经系统中起到抗抑郁效应的途径之一(Czeh et al.,the journal of the European College ofNeuropsychopharmacology 23,171-185,2013)。这些证据表明,除了神经元,胶质细胞在精神疾病中也扮演了不容忽视的角色。
内向整流型钾离子通道(inward rectifier-type potassium channel,Kir)是指超级化激活的钾离子通道。已报道的内向整流型钾离子通道有七个蛋白家族成员(Kir1ˉKir7)。同类Kir通道因为存在RNA剪接的差异(splicing variance)又可分为多种亚型,在心脏,肾脏,神经系统等多种组织和器官中分布。Kir4.1(也称为potassium voltage-gatedchannel subfamily J member 10,Kcnj10)是内向整流型钾离子通道的家族成员之一。在神经系统中,Kir4.1在胶质细胞特异性表达。胶质细胞中的Kir4.1可允许钾离子在细胞膜上通过,通过调节神经细胞周围外液的钾离子浓度,转运胞外过多的钾离子来缓冲细胞外环境,达到控制静息膜电位水平,维持神经系统的内稳态,保持神经系统的正常生理活性的作用。Kir4.1的功能异常将对胶质细胞和神经元的功能产生很大的影响,进而表现出多种神经系统病变。在哺乳动物中,Kir4.1的蛋白序列和编码核酸序列都很保守。
本领域已经具有一些常用的抗抑郁药物,但这些药物通常在比较长的一段时间后才能见效。而且导致抑郁症的病理机制还未完全被认识。本领域还需要新的,或是起效更快速、使用剂量更安全的治疗抑郁症的方法和药物。
发明内容
本发明人经过深入研究,首次发现外侧缰核的星形胶质细胞中的Kir4.1是抑郁症的一个至关重要的调节因子,运用分子、行为和电生理等手段,确定了在外侧缰核中,星形胶质细胞表达的Kir4.1,以一种紧密包绕缰核神经元胞体的方式存在于外侧缰核中,调节胞外钾平衡,改变外侧缰核神经元的发放特性,导致外侧缰核过度活跃,进而调节抑郁表型。本发明人还发现和证明了多种可以阻断外侧缰核中Kir4.1功能的试剂,由此提供了通过抑制Kir4.1的活性来治疗(抑制)抑郁症的方法和药物。
具体的,本发明提供了通过抑制Kir4.1的活性来治疗患者的抑郁症的方法。本发明还提供了Kir4.1抑制剂在制备治疗患者的抑郁症的药物中的用途。
Kir4.1抑制剂是指能够使得Kir4.1通道的活性下降或者丧失的试剂。Kir4.1通道的活性是指允许钾离子在细胞膜上通过的活性。Kir4.1通过调节神经细胞周围外液的钾离子浓度,转运胞外过多的钾离子来缓冲细胞外环境,达到控制静息膜电位水平,影响神经系统的生理活性。Kir4.1抑制剂包括能够使得Kir4.1通道的活性下降或者丧失的化合物、复合物或混合物,以及在抑制Kir4.1活性的方法(含外科手术方法)中使用的制剂等。
在本发明的通过抑制Kir4.1的活性来治疗抑郁症的方法和Kir4.1抑制剂在制备治疗抑郁症的药物中的用途中,所述Kir4.1抑制剂是指能够影响Kir4.1蛋白的活性从而影响Kir4.1通道的活性的试剂。所述试剂包括小分子化合物或复合物,或是蛋白、核酸等大分子活性成分,例如与Kir4.1蛋白结合的拮抗剂如抗体,或是影响Kir4.1蛋白的表达水平的核酸等。这些蛋白或核酸可通过本领域公知的技术,例如与合适的表达载体结合传送到目标组织或细胞发挥作用。
在本发明的其中又一个方面,Kir4.1抑制剂是特异性抑制Kir4.1的抑制剂。特异性Kir4.1抑制剂一般是指,所述Kir4.1抑制剂对其它Kir蛋白没有抑制活性,或是对其它Kir蛋白的抑制活性显著小于对Kir4.1的抑制活性,例如对其它Kir蛋白的抑制活性为对Kir4.1的抑制活性的50%以下,优选20%以下,更优选5%以下。
在本发明的其中又一个方面,所述Kir4.1抑制剂为干扰Kir4.1表达的干扰RNA或其前体。RNA干扰(RNAi)是通过双链RNA(double-stranded RNA,dsRNA)来诱发同源mRNA高效特异性降解,从而使得目标基因的表达降低甚至消除。在本发明中,干扰RNA可包括小干扰RNA(Small interfering RNA,siRNA)、短发夹RNA(shRNA)和/或微小RNA(miRNA)。在体内给予干扰RNA的一种方式是通过给予siRNA的前体shRNA实现,例如包括两个短反向重复序列的短发夹RNA。将siRNA序列作为“短发夹”克隆进质粒载体中。当送入动物体内时,该发夹序列被表达出来,形成一个“双链RNA”,利用细胞内的Dicer酶,生成相应的siRNA,发挥RNAi作用。
在本发明的其中又一个方面,可用于本发明的干扰RNA或其前体具有与目标Kir4.1 mRNA的片段具有相同或互补,或90%以上相同或互补的序列。例如,在本发明的其中又一个方面,所述干扰Kir4.1表达的干扰RNA或其前体具有以下序列:
5’-GGACGACCTTCATTGACAT-3’(SEQ ID No.1);
5’-GCTACAAGCTTCTGCTCTTCT-3’(SEQ ID No.2);
5’-GCTCTTCTCGCCAACCTTTAC-3’(SEQ ID No.3);
5’-CCGGAACCTTCCTTGCAAA-3’(SEQ ID No.4);
5’-GCGTAAGAGTCTCCTCATTGG-3’(SEQ ID No.5);或
5’-GCCCTTAGTGTGCGCATTA-3’(SEQ ID No.6)。
上述干扰RNA或其前体针对具有SEQ ID No.7(即Genebank编号NM_031602.2的序列中的CDS区序列)的大鼠Kir4.1 mRNA序列的对应mRNA片段的序列,即目标大鼠Kir4.1mRNA片段的序列与所述SEQ ID No.1-6的干扰RNA的序列相同或互补,或90%以上序列相同或互补。
本领域的技术人员可以理解和得到在其它哺乳动物(如人、小鼠等)中对应的所述Kir4.1的干扰RNA序列片段。
在本发明的其中又一个方面,所述Kir4.1抑制剂为钾离子通道活性下降或丧失的突变型Kir4.1蛋白或其编码序列。突变型蛋白可与正常蛋白发生竞争性作用,从而降低正常蛋白发挥的活性。突变型蛋白可通过给予在目标组织或细胞可表达的载体(载体上携带可表达的突变型蛋白基因和/或其表达因子)等方式在目标组织或细胞表达。在本发明的其中又一个方面,所述突变型Kir4.1蛋白为在Kir4.1蛋白的核孔区,例如在对应Kir4.1序列为SEQ ID No.8的氨基酸序列中第130-132位的GYG发生突变的突变型Kir4.1蛋白,例如GYG突变为AAA的突变型Kir4.1蛋白。具有序列为SEQ ID No.8的氨基酸序列的Kir4.1是大鼠的Kir4.1(NP_113790.2)。本领域的技术人员可以根据已有的报道(例如Hiroshi et al.,2010.Physiological Reviews 90,291-366,2010),以及对Kir4.1的序列的保守性的理解,得到在其它哺乳动物中对应的所述Kir4.1的核孔区突变位点的序列以及其它突变方式。
在本发明的其中又一个方面,所述Kir4.1抑制剂为Kir4.1的特异性抗体,包括多克隆抗体或单克隆抗体。。
本领域存在一些个别的报道某些化合物可用于抗抑郁。例如,丁螺环酮(buspirone),米安色林(mianserin),氟西汀(fluoxetine),舍曲林(sertraline),氟伏沙明(fluvoxamine)或去甲替林(nortriptyline)。但在这些报道中,其抗抑郁的机制与本发明发现的机制,即外侧缰核的星形胶质细胞表达的Kir4.1通过抑制外侧缰核神经元的异常发放,特别是簇状放电的异常发放来抑制抑郁症,完全不同。在不破坏本发明的创新性的情况下,在本发明的其中一个方面,本发明提供的上述通过抑制Kir4.1的活性来治疗抑郁症的方法和Kir4.1抑制剂在制备治疗抑郁症的药物中的用途中,所述Kir4.1抑制剂不为丁螺环酮(buspirone),米安色林(mianserin),氟西汀(fluoxetine),舍曲林(sertraline),氟伏沙明(fluvoxamine)或去甲替林(nortriptyline)。在不破坏本发明的创新性的情况下,在本发明的其中一个方面,本发明提供的上述通过抑制Kir4.1的活性来治疗抑郁症的方法和Kir4.1抑制剂在制备治疗抑郁症的药物中的用途中,所述Kir4.1抑制剂不为选择性五羟色胺重摄取抑制剂(SSRIs)或三环类抗抑郁药(TCAs)。
在本发明中,所述抑郁症可以特别指“外侧缰核介导的抑郁症”。本申请的发明人发现了外侧缰核的神经元的异常发放,特别是簇状放电的异常发放在抑郁症的产生中具有重要作用。本申请的发明人还发现了外侧缰核的星形胶质细胞中的Kir4.1是抑郁症的一个至关重要的调节因子,并发现和证明了多种可以阻断外侧缰核中Kir4.1功能的试剂,由此提供了通过抑制外侧缰核的星形胶质细胞中的Kir4.1的活性来治疗(抑制)抑郁症的方法和药物。这是本领域已知的治疗抑郁症的机制和药物未能针对的抑郁症病理机制和治疗抑郁症的脑部靶组织或其分子水平上的靶目标。因此,本发明提供的方法和药物特别适合用于在其它抗抑郁方法和药物不起效的抑郁症患者中使用。
在本发明的其中一个方面,本发明提供的方法和药物为在外侧缰核中局部起效的方法和药物。对于用于神经组织的药物,特别是脑部神经组织,例如外侧缰核来说,将药物的作用限定在目标组织是有益的。用于LHb的方法或药物需要考虑该方法或药物是否能够在LHb发挥药物的有效性,包括药物是否能到达LHb,以及在LHb中是否能达到起效的浓度等。在本发明中,所述药物可以为在外侧缰核局部给药的剂型。可以通过局部给药的方式来达到将药物作用限定在目标组织,例如通过将药物制成可通过套管植入外侧缰核局部给药的剂型。又例如,将药物制成植入组织后缓释的剂型等。另外还可将上述药物制成组织特异性的靶向药物递送系统的形式。例如可以通过将具有抑制簇状放电功能的小分子化合物或生物活性分子(核酸如蛋白编码DNA或mRNA分子、蛋白如抗体等)与能够特异性结合在外侧缰核特异性表达的蛋白结合的抗体连接形成能够识别和结合外侧缰核的细胞的复合分子。
在本发明的其中一个方面,在本发明提供的上述在通过在外侧缰核中局部抑制Kir4.1的活性来治疗抑郁症的方法和Kir4.1抑制剂在制备在外侧缰核中局部起效的治疗抑郁症的药物的用途中,所述Kir4.1抑制剂也可为选择性的五羟色胺重摄取抑制剂(丁螺环酮(buspirone),米安色林(mianserin),氟西汀(fluoxetine),舍曲林(sertraline)或氟伏沙明(fluvoxamine)等)和三环类抗抑郁药(去甲替林(nortriptyline)等)。
本发明还提供了用于治疗抑郁症的药物组合物。本发明提供新的治疗抑郁症的药物组合物,其包含治疗有效量的Kir4.1抑制剂。所述Kir4.1抑制剂如前面所定义。
在其中一个方面,本发明提供的用于治疗抑郁症的药物组合物中,所述Kir4.1抑制剂为干扰Kir4.1表达的干扰RNA或其前体,其具有与目标Kir4.1 mRNA的片段相同或互补,或90%以上相同或互补的序列。优选的,所述干扰RNA或其前体具有以下序列:
5’-GGACGACCTTCATTGACAT-3’(SEQ ID No.1);
5’-GCTACAAGCTTCTGCTCTTCT-3’(SEQ ID No.2);
5’-GCTCTTCTCGCCAACCTTTAC-3’(SEQ ID No.3);
5’-CCGGAACCTTCCTTGCAAA-3’(SEQ ID No.4);
5’-GCGTAAGAGTCTCCTCATTGG-3’(SEQ ID No.5);或
5’-GCCCTTAGTGTGCGCATTA-3’(SEQ ID No.6)。
在其中一个方面,本发明提供的用于治疗抑郁症的药物组合物中,所述Kir4.1抑制剂为钾离子通道活性下降或丧失的突变型Kir4.1蛋白或其编码序列,所述突变型Kir4.1蛋白优选为在Kir4.1蛋白的核孔区,对应Kir4.1序列为SEQ ID No.8的氨基酸序列中第130-132位的GYG发生突变的突变型Kir4.1蛋白,例如突变为AAA的突变型Kir4.1蛋白。
在其中一个方面,本发明提供的用于治疗抑郁症的药物组合物中,所述Kir4.1抑制剂为Kir4.1的特异性抗体,包括多克隆抗体或单克隆抗体。
在不破坏本发明的创新性的情况下,在本发明的其中一个方面,本发明提供的上述药物组合物中,所述Kir4.1抑制剂不为丁螺环酮(buspirone),米安色林(mianserin),氟西汀(fluoxetine),舍曲林(sertraline),氟伏沙明(fluvoxamine)或去甲替林(nortriptyline)。在不破坏本发明的创新性的情况下,在本发明的其中一个方面,本发明提供的上述治疗抑郁症的药物组合物中,所述Kir4.1抑制剂不为选择性五羟色胺重摄取抑制剂(SSRIs)或三环类抗抑郁药(TCAs)。
本发明提供的药物组合物特别适合用于在其它抗抑郁方法和药物不起效的抑郁症患者中使用。
在本发明的其中一个方面,本发明提供的药物组合物为在外侧缰核中局部起效的本发明提供的药物组合物。例如为在外侧缰核局部给药的剂型。在本发明的这一个方面,所述Kir4.1抑制剂也可为选择性地五羟色胺重摄取抑制剂(氟西汀(fluoxetine),舍曲林(sertraline),氟伏沙明(fluvoxamine),丁螺环酮(buspirone),米安色林(mianserin)等)和三环类抗抑郁药(去甲替林(nortriptyline)等)。
本发明还提供了一种抑郁症动物模型,优选为大鼠或小鼠。本发明所述抑郁症动物模型具有抑郁症特征,其外侧缰核中Kir4.1是高表达的。
本发明还提供了采用上述动物模型筛选用于治疗抑郁症的潜在物质的方法,包括步骤:
(1)给抑郁症动物模型施用待筛选的测试物;和
(2)观察所述抑郁症动物模型中的抑郁症的相关症状和/或指标,并与对照组进行比较。
其中,如果所述抑郁症动物模型中抑郁症的相关症状有显著改善,则表示该测试物是可用于治疗抑郁症潜在物质。
在本发明的其中又一个方面,提供了一种筛选用于治疗抑郁症的潜在物质的方法,其特征在于,包括步骤:
(1)在测试组中,向体外检测体系中加入待检测的测试物;和
(2)检测所述测试组的体外检测体系中Kir4.1的表达水平和/或活性,并与阴性对照组进行比较。
其中,如果与加入了阴性对照组相比,测试组中Kir4.1的表达水平显著下降,和/或Kir4.1的通道功能显著下降,则表示所述测试物是预防和/或治疗抑郁症的潜在物质。
在本发明的其中又一个方面,所述筛选用于治疗抑郁症的潜在物质的方法还包括以下一个或多个步骤:
进一步测试所述潜在物质对神经元簇状发放的影响;和/或
将所述潜在物质施用于动物模型,观察其对抑郁症症状的影响;
在测试其对神经元簇状发放的影响时,如果与阴性对照组(或空白对照组)相比,加入或施用所述测试物的测试组中神经元簇状发放比例显著降低,则表示该测试物是治疗抑郁症的潜在物质。
术语
“Kir4.1”,或“内向整流型钾离子通道(inward rectifier potassium channel)Kir4.1”,也称为potassium voltage-gated channel subfamily J member 10(Kcnj10),是内向整流型钾离子通道的家族成员之一。胶质细胞中的Kir4.1可允许钾离子在细胞膜上通过,通过调节神经细胞周围外液的钾离子浓度,转运胞外过多的钾离子来缓冲细胞外环境,达到控制静息膜电位水平,维持神经系统的内稳态,保持神经系统的正常生理活性的作用。在哺乳动物中,Kir4.1的蛋白序列和编码核酸序列都很保守。人类的Kir4.1蛋白(NP_002232)的编码基因是KCNJ10(Ensembl:ENSG00000177807)。大鼠Kir4.1蛋白(NP_113790)的编码基因是KCNJ10(Ensembl:ENSMUSG00000044708)。
本发明中,“治疗”包括:改良、减轻、减少或预防与抑郁症相关的症状的进行中的过程或结果;改善与抑郁症相关的症状的进行中的过程或结果;使处于导致特定机体功能损伤的疾病或病症中的机体功能正常化的进行中的过程或结果;或者引发疾病的一种或多种临床可测定的参数改善的进行中的过程或结果。在一个实施方案中,治疗目的是预防或减慢(减轻)不希望的生理情况、病症或疾病,或获得有益的或期望的结果。该结果可以是,例如医学的、生理学的、临床的、物理治疗、职业治疗,面向保健人员或患者;或本领域理解为“生活品质”或日常生活活动的参数。本发明中,有益的或期望的临床结果包括但不限于,减轻症状;减小/缩小该情况、病症或疾病的程度;稳定(即非恶化)该情况、病症或疾病的状态;延迟该情况、病症或疾病的开始或减慢其进展;改善或缓和该情况、病症或疾病;和减轻(无论部分或总体)、无论可检测出的或不可检测出的;或增强或改善该情况、病症或疾病。在一个实施方案中,治疗包括引发临床有效响应而没有过度水平的副作用。在一个实施方案中,治疗也包括与如果不接受治疗的预期的存活期相比延长存活期。在一个实施方案中,治疗指给药药物或对患者执行医疗程序。本发明中,治疗可以是预防(防止),治愈虚弱或病,或改良患者的临床情况,包括降低病程或疾病严重度,或主观改善患者的生活品质或延长患者的存活期。
术语“簇状发放”,或“簇状放电”,是指神经元在放电过程中同时产生两个或两个以上锋电位的放电模式。
抑制簇状放电是指抑制簇状放电的发放程度,包括减少簇状放电的频率或放电过程中簇内峰电位的个数,降低簇状放电的强度,甚至是消除簇状放电的发生。
术语“单个发放”,或“单个放电”,是神经元在放电过程中每次发放一个锋电位的放电模式。
附图说明
图1显示了抑郁大鼠外侧缰核中Kir4.1表达和功能的上调。(A,B)Western Blot实验显示cLH大鼠和LPS(脂多糖)诱导的抑郁大鼠缰核Kir4.1表达显著高于对照组。组织中大量表达的微管蛋白Tubulin被用于加载控制。蛋白表达进行定量标准化。(C,D)成年cLH大鼠(60-90天龄)而非未成年大鼠(30天龄)中,外侧缰核Ba+敏感的电流(多由Kir4.1通道介导)显著增高。(E,F)成年大鼠而非未成年cLH大鼠在习得性无助和强迫游泳行为测试中表现出了显著的抑郁表型。所有数据均表示为平均值±SEM。*P<0.05,**P<0.01,***P<0.001,****P<0.0001与对照组相比。N.S.表示差异不显著。其他的图标相同。
图2显示了外侧缰核中Kir4.1表达在星形胶质细胞的突起上并紧密包裹神经元胞体。(A)免疫荧光双标染色显示Kir4.1免疫阳性信号包绕在神经元的胞体周围。(B)注射腺相关病毒AAV2/1-CaMKII-EGFP-Cre到Kir4.1条件性敲除小鼠的外侧缰核,条件性敲除外侧缰核神经元中的Kir4.1,依然可以看到Kir4.1包绕神经元胞体的现象;但注射AAV2/5-GFAP-EGFP-Cre病毒敲除外侧缰核星形胶质细胞中的Kir4.1后包绕现象消失。(C)免疫电镜观测到标记了金颗粒的Kir4.1信号分布在神经元胞体的周围。(D)全细胞膜片钳电生理技术记录到星形胶质细胞而非神经元上有Ba2+敏感的Kir4.1电流。所有数据均表示为平均值±SEM。****P<0.0001与对照组相比。N.S.表示差异不显著。
图3Kir4.1在外侧缰核和海马中的细胞定位。(A)外侧缰核中Kir4.1与神经元标志物NeuN,星形胶质细胞标志物GFAP和S100b的共定位;(B)海马CA1区中Kir4.1与神经元标志物NeuN,星形胶质细胞标志物GFAP和S100b的共定位。最下方两幅图显示Kir4.1抗体被Kir4.1胞外肽段抗原吸附后在外侧缰核和海马CA1中,均检测不到阳性信号,提示所用抗体是特异性针对Kir4.1的。
图4外侧缰核细胞外钾浓度与神经元活性高度相关且被星形胶质细胞Kir4.1所调节。(A-C)胞外灌流含BaCl2的人工脑脊液阻断星胶上Kir4.1通道,可使外侧缰核中簇状发放(burst)和基础发放(tonic)神经元的静息膜电位显著增高,神经元表现为去极化。(D-F)神经元去极化的程度与自身发放频率成正相关。(G)BaCl2对神经元发放作用的典型图。(H)BaCl2显著降低簇状发放细胞的每分钟内簇状发放的频率。(I-K)降低胞外钾可以显著地降低神经元的静息膜电位,增加簇状发放细胞的比例。*P<0.05,**P<0.01,***P<0.001,与对照组相比。N.S.表示差异不显著。
图5星形胶质细胞Kir4.1过量表达增加神经元簇状发放细胞的比例且导致抑郁表型。(A)星形胶质细胞Kir4.1过量表达的病毒载体构建示意图。(B)免疫荧光显示外侧缰核中病毒在星形胶质细胞中过量表达Kir4.1蛋白。(C,D)Kir4.1过表达使神经元和星形胶质细胞的静息膜电位都显著下降,表现为超级化。(E)星形胶质细胞Kir4.1过量表达增加神经元簇状发放细胞的比例(F)星形胶质细胞Kir4.1过量表达增加了高频发放细胞的比例(G)星形胶质细胞Kir4.1过量表达小鼠抑郁表型检测时间轴(H,I)星形胶质细胞Kir4.1过量表达小鼠在强迫游泳和糖水偏好实验中都表现出显著的抑郁表型。所有数据均表示为平均值±SEM。*P<0.05,**P<0.01,***P<0.001,****P<0.0001与对照组相比。N.S.表示差异不显著。
图6下调LHb中Kir4.1的功能能降低神经元中簇状发放神经元的比例且有效的缓解抑郁症状。(A)用于下调Kir4.1表达水平的RNAi和下调Kir4.1功能的Kir4.1显性突变的AAV病毒载体示意图。(B)Western blot实验显示体外培养的HEK293细胞中,Kir4.1-shRNA可有效降低过表达在细胞上的Kir4.1.(C)上图显示Kir4.1功能下调大鼠进行电生理学实验和行为学检测的时间轴。下图免疫荧光显示Kir4.1显性突变病毒在外侧缰核星形胶质细胞中表达。(D)Kir4.1下调的星形胶质细胞反转点位升高。(E-G)Kir4.1下调的星形胶质细胞静息膜电位显著增高,神经元的静息膜电位也显著增高,但周围星胶是否有Kir4.1的下调,神经元的膜电位变化没有差异。(H)下调Kir4.1的表达显著降低外侧缰核簇状发放细胞比例。(I)下调Kir4.1表达及功能均显著降低cLH大鼠在强迫游泳中的不动时间。(J,K)下调Kir4.1表达及功能反转cLH在习得性无阻行为检测中的抑郁表型。(L)下调Kir4.1表达及功能显著反转cLH大鼠的糖水偏好缺失。
图7不同Kir4.1 shRNA敲除效率的体外验证。运用蛋白免疫印迹的方法,检测体外培养的HEK293细胞中外源性表达的Kir4.1,被6条针对Kir4.1不同序列的shRNA敲除的效率。
图8LHb病毒注射过表达Kir4.1或下调Kir4.1均不影响动物的运动能力。(A)AAV2/5-gfaABC1D-EGFP-Kir4.1病毒表达,LHb过表达Kir4.1后,小鼠在矿场中总的运动距离和中央区停留时间与GFP对照组相比均无显著差异。(B)AAV2/5-Kir4.1 shRNA病毒表达,LHb中Kir4.1表达下调后,cLH大鼠在旷场中的总运动距离和中央区停留时间与对照病毒相比无差异。
具体实施方式
下面将结合实施例进一步说明本发明的实质内容和有益效果,该实施例仅用于说明本发明而非对本发明的限制。下列实施例中未注明具体条件的实验方法,通常按照常规条件,例如Sambrook等人,分子克隆:实验室手册(New York:Cold Spring HarborLaboratory Press,1989)中所述的条件,或按照制造厂商所建议的条件。
实施例1材料和方法
动物材料
雄性cLH大鼠(4-12周龄),Sprague Dawley大鼠(4-12周龄)和雄性Wistar大鼠(12周龄)。cLH大鼠是一个选择性培育的具有先天习得性无助抑郁表型的抑郁症动物模型(D.Schulz,M.M.Mirrione,F.A.Henn,Neurobiol Learn Mem 93,291,Feb,2010)。本实验的cLH大鼠从美国冷泉港Malinow实验室引进。cLH大鼠饲养和繁殖如前述D.Schulz,et al,Feb,2010中描述。大鼠4只/笼,12小时的明暗周期(7am-7pm有光)。用于套管实验的cLH大鼠1只/笼饲养。成年(8-12周龄)C57BL/6小鼠被用于行为测试:4只/笼,12小时明暗周期(5am-5pm有光)。大鼠和小鼠都能够自由摄取稳定的水和食物,所有的动物实验经过浙江大学动物保护和使用委员会的批准。
病毒构建
Kir4.1的过表达病毒AAV2/5-gfaABC1D-EGFP-Kir4.1。Kir4.1过表达质粒:pZac2.1 gfaABC1D-EGFP-Kir4.1,购自于AddGene公司,货号:Plasmid#52874。并由上海泰廷生物科技有限公司包被成AAV2/5型病毒,即AAV2/5-gfaABC1D-EGFP-Kir4.1。
AAV2/1-CamKII-HI-eGFP-Cre,购自于美国宾夕法尼亚大学载体中心(Universityof Pennsylvania Vector core,Upenn,USA),Cat#:AV-1-PV2521。
Kir4.1下调病毒AAV2/5-H1-Kir4.1-shRNA-gfaABC1D-EGFP(Kir4.1的干扰RNA-shRNA:5’-GCGTAAGAGTCTCCTCATTGG-3’),对照病毒AAV2/5-H1-Luciferase-shRNA-gfaABC1D–EGFP;Kir4.1显性突变病毒AAV2/5-gfaABC1D-Kir4.1dn-2A-EGFP(Kir4.1序列的核孔区GYG突变为AAA而实现Kir4.1的显性突变)。均由由上海泰廷生物科技有限公司制备质粒和包被。
LPS(脂多糖)诱导抑郁模型
12周龄的雄性Wistar大鼠,每天腹腔注射1次LPS(500μg/kg体重),连续注射7天,最后一次注射结束后24h,进行FST测试,测试结束后3天取出缰核组织,进行表达量分析。
立体定位注射和组织学
小鼠注射病毒:小鼠腹腔注射氯胺酮(100mg/kg体重)和赛拉嗪(8mg/kg)混合液麻醉后,固定于立体定位仪上(Stoelting instruments)。每只小鼠每侧LHb注入0.1-0.2ul纯化浓缩的AAV病毒(~1013感染单位/ml),LHb立体定位坐标(前后距离Bregma:-1.7mm(AP),左右旁开±0.46mm(ML),皮层表面往下-2.56mm(DV))。使用自行拉制的玻璃微电极缓慢注入(~100-150nl/min),注射结束留针5min,然后再5min内缓慢移出注射电极。
大鼠注射病毒:大鼠腹腔注射4%戊巴比妥那(60mg/kg体重)麻醉后,固定于大鼠立体定位仪上。每只大鼠每测LHb注射入0.1-0.2ul纯化浓缩的AAV病毒(ˉ1013感染单位/ml),LHb立体定位坐标(前后距离Bregma:-3.7mm(AP),左右旁开±0.7mm(ML),皮层表面往下-4.55mm(DV))。使用自行拉制的玻璃微电极缓慢注入(~100-150nl/min),注射结束留针5min,然后再5min内缓慢移出注射电极。
术后至少14天,开展行为实验或者电生理实验。行为实验结束后检查注射位置,只使用正确注射的那些动物数据。注射了GFP标记的病毒在显微检查之前用抗体检查GFP蛋白。每个大脑的缰区切成6组连续的切片(小鼠30μm的切片,每组6片;大鼠40μm
切片,每组8-9片)。所有的切片在安装到固定片之前用Hoechst复染色。
免疫组化
首先腹腔注射4%的戊巴比妥那让动物进入深度麻醉。然后分别用PBS和冰预冷4%的PFA进行灌流固定,取出动物脑组织经4%的PFA后固定过夜。30%的蔗糖溶液脱水处理1-3天,待脑组织沉到管底,可进行冰冻切片,切片厚度为40μm,并-20度冻存于切片保护液中。抗体的浓度如下:anti-Kir4.1(1:200,Alomone labs),anti-NeuN(1:500,chemicon),anti-GFAP(1:500,Chemicon),anti-S100b(1:500,Invitrogen,ABCAM),anti-GFP(1:1000,abCaM)。Alexa Fluor488 goat anti-rabbit IgG,Alexa Fluor488 goatanti-chicken IgG,Alexa Fluor594 goat anti-mouse IgG(all 1:1000,Invitrogen),Hoechst(1:5000)荧光图片利用Olympus Fluoview FV1000和NiconA1激光共聚焦显微镜采集图像。
蛋白免疫印迹
大鼠经异氟烷麻醉后,断头取脑。冰上快速分离缰核组织并冻于液氮中。样品在匀浆液内进行组织破碎。组织匀浆物以800g在4度离心15分钟。取上清并以10000g离心15min。沉淀物即为膜蛋白组分,然后将它们溶解在匀浆液中。293TN细胞系的样品经过RIPA溶液(20mM Tris-HCl[pH 7.5]、150mM NaCl、1mM EDTA、1mM EGTA、1% NP-40、1%sodiumdeoxycholate、1mM PMSF、10μg/ml aprotinin、1μg/ml pepstatin A和1μg/ml leupeptin)裂解后,10000g离心15min取上清。用BCA法测定蛋白浓度后,以8-15μg蛋白每孔上样,10%SDS-PAGE胶进行分离,PVDF膜转印做Western印迹。所用一抗为anti-Kir4.1-introcellular(1:1000,Alomone labs),anti-tubulin(1:10000,Bio-Rad),对于免疫印迹结果的定量,使用Quantity One或image J软件进行统计分析。
电生理
出生后40-50天的大鼠或出生8周的小鼠经异氟烷麻醉后,用20ml冰冷充氧的切片液进行灌流。快速断头取出大脑,放进充氧的切片液中。随后利用Leica振动切片机在充氧的冰冷的切片液中,进行350μm的冠状切面切片。缰核脑片在充氧的34℃的ACSF(118mMNaCl,2.5mM KCl,26mM NaHCO3,1mM NaH2PO4,10mM glucose,1.3mM MgCl2 and 2.5mMCaCl2,gassed with 95% O2 and 5% CO2)中,恢复两小时后转移到室温进行记录。
外侧缰核脑片的膜片钳记录采用Axon Multiclamp 700B放大器,在32±1℃环境下,在装配红外微分干涉相差光学镜头的Olympus显微镜下,进行记录。星形胶质细胞记录电极的阻抗为7-10MΩ,神经元记录电极阻抗为4-6MΩ。所有细胞均在全细胞模式下记录。根据所记录细胞的形态,记录电生理特性判断是神经元(15-20微米直径)还是星形胶质细胞(5-10微米直径)。神经元记录参数包括细胞膜电位,膜阻抗,动作电位自发放频率(I=0记录),神经元电压电流曲线以及神经元输入输出兴奋性。星形胶质细胞记录参数包括膜电位,膜阻抗,电压电流曲线(电压钳下从-130mV至40mV电压,每次记录增加10mV,记录各个电位下的电流反应)。Kir4.1电流计算方法:由正常人工脑脊液中记录得到的电压电流曲线减去加入氯化钡(100Mm,该浓度特异性拮抗Kir4.1电流)的电压电流曲线。数据经过2kHz过滤后使用Digidata 1322A在10kHz下采样记录。数据使用pClamp 10软件进行分析。
行为学实验
习得性无助实验
实验是对同窝cLH或野生型SD大鼠进行。分两部分进行:“训练部分”包括在刺激室(coulbourn仪器)中,120次不可避免的、不受控的0.8mA足底电刺激超过40分钟,随机刺激持续时间和范围从5秒到15秒间隔刺激时间,总刺激持续时间为20分钟。“测试部分”训练24小时后进行,习得无助表型是由一个杆压迫任务进行评估,期间一个光照指示杆被放入刺激室中。这一部分包括15次可逃避的0.8mA强度电刺激和24s实验间隔。每次刺激持续了60秒,但是能被杆压迫终止。超过10次失败被定义为“习得无助”(LH);少于5次失败为“非习得无助”(NLH)。
强迫游泳测试
实验在正常日光灯下进行。小鼠强迫游泳圆柱形容器的直径为12cm,高25cm。测试水深为14cm,水温23-24℃。摄像头从侧边记录小鼠在6min内的游泳情况。采用双盲方式统计小鼠游泳6min内后4min的不动时间(动物的漂浮姿势或者四肢完全没有活动的时间)。
糖水偏好测试
实验小鼠单独饲养1周,然后连续2天给予小鼠两瓶普通水,之后两天将水换为两瓶2%的蔗糖水进行训练。训练结束后,给予动物一瓶普通水和一瓶2%的蔗糖水进行测试,每12小时交换一次水瓶的位置,每24小时记录一次水和糖水的消耗量(对水瓶称重),共记录48小时。
统计分析
所有的数据都以平均值±SEM。对于所有的行为数据,采用two-tailed Student'st-tests或Mann-Whitney test。
实施例2抑郁大鼠缰核中Kir4.1表达和活性上调
如图1所示。在动物体内观察抑郁大鼠外侧缰核中Kir4.1表达和活性。分别在先天抑郁(cLH)大鼠和另一种抑郁症的动物模型——脂多糖(LPS)诱导的大鼠抑郁模型。脂多糖(LPS)诱导的大鼠抑郁模型是对3月龄的雄性Wistar大鼠,连续7天每天腹腔注射500ug/kg的LPS,最后一次注射后24小时,检测到大鼠在强迫游泳中的不动性显著增加。
图1中的A和B是蛋白免疫印迹实验结果,显示cLH大鼠和LPS(脂多糖)诱导的抑郁大鼠的外侧缰核中的Kir4.1表达显著高于对照组。图1中的A显示cLH大鼠缰核膜组分中Kir4.1蛋白表达水平比对照增高了1.75倍。图1中的B显示外侧缰核中Kir4.1的表达水平在LPS诱导的抑郁大鼠中也显著增高(1.87倍)。
为了确定抑郁大鼠中Kir4.1的功能是否也增强,运用全细胞膜片钳记录技术检测了cLH和SD大鼠LHb脑片中星形胶质细胞和神经元的变化。根据星形胶质细胞胞体较小(5-10μm)的形态学特征,和一系列电生理学的特征来分离星形胶质细胞。这些电生理特征包括:相对超级化的静息膜电位(-74±1mV);低的输入阻抗Ri(47±6MΩ);线性的电流-电压曲线关系和去极化电流不能使它产生动作电位。运用Ba2+去选择性地阻断Kir通道的电流,进而分离出Kir4.1的电流。电生理记录发现,在成年(60-90天)cLH大鼠LHb的星形胶质细胞中,Ba2+敏感的Kir4.1电流几乎是对照SD组电流的两倍(图1的C)。同时,在未成年的cLH大鼠中Kir4.1的电流没有增高(图1的D),且未成年大鼠的强迫游泳的不动时间和习得性无助实验中为了逃脱电击而按指示杆的次数与对照组都没有差异(图1的E和F)。这显示,Kir4.1的过表达与抑郁症状的起始有关联。
实施例3 Kir4.1在外侧缰核的表达分布特性
运用免疫荧光双标的方法,发明人出乎意料地首次发现Kir4.1以包绕在神经元的胞体上表达的方式在外侧缰核中分布。而在海马中,Kir4.1的表达主要是星形状的。(如图3所示)
图2显示了外侧缰核中Kir4.1表达在星形胶质细胞的突起上并紧密包裹神经元胞体。
图2的A表示,免疫荧光双标染色显示Kir4.1免疫阳性信号包绕在神经元的胞体周围。
另外,通过运用条件性敲除的方法来来观察和判断Kir4.1的表达。通过注射腺病毒AAV2/1-CaMKII-EGFP-Cre到Kir4.1条件性敲除小鼠的外侧缰核来实现条件性敲除。如图2的B所示,条件性敲除外侧缰核神经元中的Kir4.1,依然可以看到Kir4.1包绕神经元胞体的现象;但注射AAV2/5-GFAP-EGFP-Cre病毒来敲除外侧缰核星形胶质细胞中的Kir4.1后,包绕现象消失。
运用免疫电镜的方法也检测到Kir4.1免疫金颗粒在神经元细胞膜周围分布,而在神经元突触部位较少(如图2的C所示)。
电生理证据也同样地发现Ba2+敏感的Kir4.1电流主要在星形胶质细胞被记录到而非神经元(如图2的D所示)。
图3给出的是Kir4.1在外侧缰核和海马中的细胞定位。图3的A显示外侧缰核中Kir4.1与神经元标志物NeuN,星形胶质细胞标志物GFAP和S100b的共定位;图3的B显示海马CA1区中Kir4.1与神经元标志物NeuN,星形胶质细胞标志物GFAP和S100b的共定位。图3最下方两幅图显示Kir4.1抗体被Kir4.1胞外肽段抗原吸附后,在外侧缰核和海马CA1中均检测不到阳性信号,提示所用抗体是特异性针对Kir4.1的。
以上结果均说明LHb中星形胶质细胞突起上的Kir4.1,主要以紧密包绕在神经元胞体周围的表达形式存在。
实施例4Kir4.1对神经元活性的调节作用
运用膜片钳电生理记录的方法对Kir4.1对神经元活性的调节作用进行研究。首先通过胞外灌流含有BaCl2的人工脑脊液,阻断Kir4.1缓冲钾离子的功能。发现BaCl2处理能显著去极化除了无自发放的细胞外的所有LHb神经元。如图4的A-C所示,胞外灌流含BaCl2的人工脑脊液阻断星胶上Kir4.1通道,可使外侧缰核中簇状发放(burst)和基础发放(tonic)神经元的静息膜电位显著增高,神经元表现为去极化。这与Nernst方程推算的增多胞外钾浓度降低,引起神经元的膜电位超级化相一致。且BaCl2处理对LHb自发放细胞去极化的程度与细胞自身发放频率正相关(如图4的D-F所示)。BaCl2处理长时间过度兴奋簇状发放神经元,使得神经元处于强直状态,最终使神经元簇状发放数量显著降低(如图4的G-H所示)。
实施例5Kir4.1通过调节胞外钾浓度而引起神经元和星形胶质细胞的电生理功能的变化
在正常大鼠的LHb脑片上孵育钾浓度下降(2.75mM-1.4mM)的人工脑脊液,这使得神经元的静息膜电位超级化了10.3mV(如图4的I,J所示),且使簇状发放细胞比例从8%增高到了23%(如图4的K所示)。这些结果显示Kir4.1的过表达通过增加胞外钾离子的清除而超级化神经元,引发簇状发放。
实施例6动物体内实验显示在外侧缰核过表达Kir4.1引发抑郁症症状
构建Kir4.1过量表达质粒pAAV-gfaABC1D-Kir4.1-2A-EGFP和Kir4.1显性突变质粒pAAV-gfaABC1D-Kir4.1dn-2A-EGFP(Kir4.1序列的核孔区GYG突变为AAA而实现Kir4.1的显性突变)。
质粒构建:
1.首先构建pAAV-Ubi-Kir4.1-2A-EGFP质粒,用pAAV-Ubi-CaMKIIb-2A-EGFP质粒(Li et al.,2013)为模板,用表1中引物pAAV-ubi Fusion Fw和pAAV-ubi Fusion Rev扩增出一个线性化载体;再用pZac2.1-gfaABC1D-EGFP-Kir4.1(购自AddGene,Plasmid#52874)为模板,用pAAV-ubi-Kir4.1 Fusion Fw和Rev扩增出kir4.1的片段。以上获得的两个线性化片段重组成pAAV-Ubi-Kir4.1-2A-EGFP。
表1
2.构建pAAV-Ubi-Kir4.1dn-2A-EGFP质粒:以pAAV-Ubi-Kir4.1-2A-EGFP质粒为模板,用表2中的引物Kir4.1 GYG-AAA Fusion Fw和Kir4.1 GYG-AAA Fusion Rev扩增出一个线性化的突变片段,然后自身重组成完整的AAA突变的pAAV-Ubi-Kir4.1dn-2A-EGFP质粒。
表2
3.构建pAAV-gfaABC1D-Kir4.1dn-2A-EGFP质粒,方法是:以pZac2.1-gfaABC1D-EGFP-Kir4.1质粒为模板,运用表3所示的引物pZac2.1 gfaABC1D Fusion Rev和pZac2.1gfaABC1D Fusion Fw,扩增出EGFP-Kir4.1缺失的pZac2.1 gfaABC1D线性片段;再以pAAV-Ubi-Kir4.1dn-2A-EGFP质粒为模板,Kir4.1dn-2A-eEGFP Fusion Rev和Kir4.1dn-2A-eEGFP Fusion Fw为引物,扩增出Kir4.1dn-2A-EGFP片段。以上两个线性化片段同源重组后可获得pAAV-gfaABC1D-Kir4.1dn-2A-EGFP质粒。
表3
运用立体定位注射的方法,在外侧缰核注射pAAV-gfaABC1D-Kir4.1-2A-EGFP,表达14天,使Kir4.1在LHb星形胶质细胞里过表达。图5的A显示用于在星形胶质细胞过量表达Kir4.1的病毒载体构建示意图。图5的B是免疫荧光图,显示外侧缰核中病毒在星形胶质细胞中过量表达Kir4.1蛋白。采用全细胞膜片钳记录过表达Kir4.1的LHb脑片,发现其中的星形胶质细胞和神经元的静息膜电位都显著超级化(如图5的D和E所示),且神经元簇状发放的比例也显著高于注射对照GFP病毒组(如图5的F,G所示)。同时Kir4.1在LHb的过表达也显著增高了小鼠在强迫游泳中的不动性,显著降低了小鼠对糖水的偏好性(如图5的H,I所示),而小鼠的运动能力没有发生改变(图8的A所示)。这些实验证明LHb Kir4.1过表达小鼠表现出了典型的抑郁症状。
实施例7外侧缰核的Kir4.1表达水平的下调或Kir4.1功能逆转抑郁表型
采用AAV2/5病毒表达Kir4.1的短发夹RNA(shRNA)来下调cLH大鼠LHb Kir4.1蛋白表达水平。
将根据下表中构建的6个Kir4.1的干扰RNA的shRNA克隆到WX231-L载体(购自于上海泰廷生物科技有限公司,Cat#:WX231)上:
克隆采用的引物序列如下表:
表4引物序列
在体外培养的HEK293细胞中,分别共表达Kir4.1和所述6个shRNA。
图7是蛋白免疫印迹结果。用动物体内不表达的蛋白荧光素酶,作为阴性对照(negative control,NC)。AAV2/5-H1-Luciferase(荧光素酶)-shRNA-gfaABC1D–EGFP作为对照病毒表达NC-shRNA。Kir4.1shRNA的敲除效率为:Kir4.1-shRNA-1>Kir4.1-shRNA-4,Kir4.1-shRNA-5>Kir4.1-shRNA-2>Kir4.1-shRNA-6>Kir4.1-shRNA-3。
由上海泰廷生物科技有限公司包被,将Kir4.1-shRNA-5包装成载体为AAV2/5的病毒用于Kir4.1功能下调大鼠实验。
图6的A是用于下调Kir4.1表达水平的RNAi和下调Kir4.1功能的Kir4.1显性突变的AAV病毒载体示意图。图6的B是Western blot实验结果,显示体外培养的HEK293细胞中,Kir4.1-shRNA可有效降低过表达在细胞上的Kir4.1。
图6的C的上图显示Kir4.1功能下调大鼠进行电生理学实验和行为学检测的时间轴;下图免疫荧光显示Kir4.1显性突变病毒在外侧缰核星形胶质细胞中表达。
电生理记录AAV-Kir4.1-shRNA病毒表达的大鼠LHb脑片,表达shRNA的星形胶质细胞静息膜电位显著去极化,而表达shRNA的神经元静息膜电位与相邻的不表达shRNA的神经元一样,但比表达对照shRNA的神经元显著的超级化(如图6的D-G所示)。提示星形胶质细胞中Kir4.1的敲除对所有的神经元静息膜电位均有影响。Kir4.1 shRNA的表达,也使LHb中簇状发放的神经元比例从29%显著降低到0%(如图6的H所示)。同时,下调cLH大鼠LHbKir4.1蛋白表达水平显著反转cLH大鼠的抑郁表型:大鼠的强迫游泳不动性显著降低(如图6的I所示),习得性无助按杆行为显著增高(如图6的J,K所示),糖水偏好也显著增高(如图6的L所示),而大鼠的运动能力没有发生改变(如图8的B所示)。
另外,通过病毒表达Kir4.1的显性突变体去抑制cLH大鼠LHb Kir4.1通道的功能。图6的A显示Kir4.1显性突变病毒AAV2/5-gfaABC1D-Kir4.1dn-2A-EGFP的结构(Kir4.1序列的核孔区氨基酸序列130-132的GYG突变为AAA而实现Kir4.1的显性突变)。在LHb注射AAV-dnKir4.1-EGFP病毒都显著地反转cLH大鼠的抑郁表型:大鼠的强迫游泳不动性显著降低(如图6的I所示),习得性无助按杆行为显著增高(如图6的J,K所示),糖水偏好也显著增高(如图6的L所示)。
结论
本发明人经过深入研究,首次发现外侧缰核的星形胶质细胞中的Kir4.1是抑郁症的一个至关重要的调节因子,并运用分子、行为和电生理等手段,发现Kir4.1以一种紧密包绕缰核神经元胞体的方式存在于外侧缰核中,调节胞外钾平衡,改变外侧缰核神经元的发放特性,导致外侧缰核过度活跃,进而调节抑郁表型。本发明人还发现和证明了多种可以阻断外侧缰核中Kir4.1活性的试剂,由此提供了通过抑制Kir4.1的活性来治疗(抑制)抑郁症的方法和药物。
上面是对本发明进行的说明,不能将其看成是对本发明进行的限制。除非另外指出,本发明的实践将使用有机化学、聚合物化学、生物技术等的常规技术,显然除在上述说明和实施例中所特别描述之外,还可以别的方式实现本发明。其它在本发明范围内的方面与改进将对本发明所属领域的技术人员显而易见。根据本发明的教导,许多改变和变化是可行的,因此其在本发明的范围之内。
如无特别表示,本文中出现的温度的单位“度”是指摄氏度,即℃。
序列表
<110> 浙江大学
<120> 钾离子通道Kir4.1抑制剂治疗抑郁症的用途和药物组合物
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 19
<212> DNA
<213> 大鼠
<400> 1
ggacgacctt cattgacat 19
<210> 2
<211> 21
<212> DNA
<213> 大鼠
<400> 2
gctacaagct tctgctcttc t 21
<210> 3
<211> 21
<212> DNA
<213> 大鼠
<400> 3
gctcttctcg ccaaccttta c 21
<210> 4
<211> 19
<212> DNA
<213> 大鼠
<400> 4
ccggaacctt ccttgcaaa 19
<210> 5
<211> 21
<212> DNA
<213> 大鼠
<400> 5
gcgtaagagt ctcctcattg g 21
<210> 6
<211> 19
<212> DNA
<213> 大鼠
<400> 6
gcccttagtg tgcgcatta 19
<210> 7
<211> 1140
<212> DNA
<213> 大鼠
<400> 7
atgacatcag ttgccaaggt ctattacagc cagacgacgc agacagagag ccggccccta 60
gtggctccag gaatacgtcg gaggagggtc ctgacaaaag atggccggag caacgtgaga 120
atggagcata ttgctgacaa gcgtttcctc tacctcaagg atctatggac gaccttcatt 180
gacatgcagt ggcgctacaa gcttctgctc ttctcggcaa cctttgcagg cacttggttc 240
ctctttggcg tggtgtggta tctggtcgct gtggcccacg gggacctgtt ggagctggga 300
cctcctgcca accacacgcc ctgtgtggtg caggtgcaca cacttactgg ggccttcctc 360
ttctccctcg aatcccagac caccattggc tatggcttcc gctacatcag cgaggaatgc 420
cctctggcca ttgtgcttct cattgcacag ctcgtgctca ccaccattct ggaaatcttc 480
atcaccggaa ccttccttgc aaagattgcc cggccaaaga agagggctga gacgatccgt 540
ttcagccagc atgcggttgt ggcttaccac aacgggaagc tttgcctcat gatccgggtg 600
gccaacatgc gtaagagtct cctcattggg tgccaggtga caggcaaact gcttcaaacc 660
caccagacaa aggagggtga gaatattcgg ctcaaccagg tcaatgtgac tttccaagta 720
gacacagcct ctgatagccc ctttctcatt ctacccctga ctttctacca tgtggtagat 780
gagaccagcc ccttgaaaga tctccccctc cgcagcgggg agggtgactt cgagctcgtg 840
ctgatcctaa gtgggacggt ggagtccacc agcgccacct gtcaagttcg cacttcctat 900
ctaccggagg agatcctctg gggctacgag ttcacacctg ctatctcact gtcagccagt 960
ggcaaatacg tggctgactt cagccttttt gaccaggttg tgaaagtggc gtcccccggt 1020
ggtctccgag atagcaccgt acgttatgga gacccagaaa agctcaagtt ggaggagtca 1080
ttaagagagc aagctgaaaa ggaaggcagt gcccttagtg tgcgcattag taacgtctga 1140
<210> 8
<211> 379
<212> 蛋白质
<213> 大鼠
<400> 8
Met Thr Ser Val Ala Lys Val Tyr Tyr Ser Gln Thr Thr Gln Thr Glu
1 5 10 15
Ser Arg Pro Leu Val Ala Pro Gly Ile Arg Arg Arg Arg Val Leu Thr
20 25 30
Lys Asp Gly Arg Ser Asn Val Arg Met Glu His Ile Ala Asp Lys Arg
35 40 45
Phe Leu Tyr Leu Lys Asp Leu Trp Thr Thr Phe Ile Asp Met Gln Trp
50 55 60
Arg Tyr Lys Leu Leu Leu Phe Ser Ala Thr Phe Ala Gly Thr Trp Phe
65 70 75 80
Leu Phe Gly Val Val Trp Tyr Leu Val Ala Val Ala His Gly Asp Leu
85 90 95
Leu Glu Leu Gly Pro Pro Ala Asn His Thr Pro Cys Val Val Gln Val
100 105 110
His Thr Leu Thr Gly Ala Phe Leu Phe Ser Leu Glu Ser Gln Thr Thr
115 120 125
Ile Gly Tyr Gly Phe Arg Tyr Ile Ser Glu Glu Cys Pro Leu Ala Ile
130 135 140
Val Leu Leu Ile Ala Gln Leu Val Leu Thr Thr Ile Leu Glu Ile Phe
145 150 155 160
Ile Thr Gly Thr Phe Leu Ala Lys Ile Ala Arg Pro Lys Lys Arg Ala
165 170 175
Glu Thr Ile Arg Phe Ser Gln His Ala Val Val Ala Tyr His Asn Gly
180 185 190
Lys Leu Cys Leu Met Ile Arg Val Ala Asn Met Arg Lys Ser Leu Leu
195 200 205
Ile Gly Cys Gln Val Thr Gly Lys Leu Leu Gln Thr His Gln Thr Lys
210 215 220
Glu Gly Glu Asn Ile Arg Leu Asn Gln Val Asn Val Thr Phe Gln Val
225 230 235 240
Asp Thr Ala Ser Asp Ser Pro Phe Leu Ile Leu Pro Leu Thr Phe Tyr
245 250 255
His Val Val Asp Glu Thr Ser Pro Leu Lys Asp Leu Pro Leu Arg Ser
260 265 270
Gly Glu Gly Asp Phe Glu Leu Val Leu Ile Leu Ser Gly Thr Val Glu
275 280 285
Ser Thr Ser Ala Thr Cys Gln Val Arg Thr Ser Tyr Leu Pro Glu Glu
290 295 300
Ile Leu Trp Gly Tyr Glu Phe Thr Pro Ala Ile Ser Leu Ser Ala Ser
305 310 315 320
Gly Lys Tyr Val Ala Asp Phe Ser Leu Phe Asp Gln Val Val Lys Val
325 330 335
Ala Ser Pro Gly Gly Leu Arg Asp Ser Thr Val Arg Tyr Gly Asp Pro
340 345 350
Glu Lys Leu Lys Leu Glu Glu Ser Leu Arg Glu Gln Ala Glu Lys Glu
355 360 365
Gly Ser Ala Leu Ser Val Arg Ile Ser Asn Val
370 375
Claims (3)
1.Kir4.1抑制剂在制备治疗抑郁症的药物中的用途,其中所述Kir4.1抑制剂为:
(1)干扰Kir4.1表达的干扰RNA或其前体,其中,所述干扰RNA或其前体为针对Kir4.1的对应序列为SEQ ID No.7的mRNA序列中以下序列片段的干扰RNA或其前体:
5’-GGACGACCTTCATTGACAT-3’(SEQ ID No.1);
5’-GCTACAAGCTTCTGCTCTTCT-3’(SEQ ID No.2);
5’-CCGGAACCTTCCTTGCAAA-3’(SEQ ID No.4);
5’-GCGTAAGAGTCTCCTCATTGG-3’(SEQ ID No.5);或
5’-GCCCTTAGTGTGCGCATTA-3’(SEQ ID No.6);
其中,所述干扰Kir4.1表达的干扰RNA或其前体为采用以下引物对克隆而得的shRNA:
或
(2)钾离子通道活性下降或丧失的突变型Kir4.1蛋白或其编码序列,其中所述Kir4.1抑制剂为在Kir4.1蛋白的核孔区,对应Kir4.1序列为SEQ ID No.8的氨基酸序列中第130-132位的GYG突变为AAA的突变型Kir4.1蛋白。
2.治疗抑郁症的药物组合物,其中包含Kir4.1抑制剂,其中所述Kir4.1抑制剂为干扰Kir4.1表达的干扰RNA或其前体,所述干扰RNA或其前体为针对Kir4.1的对应序列为SEQ IDNo.7的mRNA序列中以下序列片段的干扰RNA或其前体:5’-GCGTAAGAGTCTCCTCATTGG-3’(SEQID No.5),
其中,所述干扰Kir4.1表达的干扰RNA或其前体为采用以下引物对克隆而得的shRNA:
3.一种核酸分子,其为针对Kir4.1的对应序列为SEQ ID No.7的mRNA序列中以下序列片段的干扰RNA或其前体:
5’-GCGTAAGAGTCTCCTCATTGG-3’(SEQ ID No.5),
其中,所述干扰Kir4.1表达的干扰RNA或其前体为采用以下引物对克隆而得的shRNA:
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210060456.1A CN115006534B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道Kir4.1抑制剂治疗抑郁症的用途和药物组合物 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710322245X | 2017-05-09 | ||
CN201710322245 | 2017-05-09 | ||
CN201810432839.0A CN108853505B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道抑制剂治疗抑郁症的用途和药物组合物 |
CN202210060456.1A CN115006534B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道Kir4.1抑制剂治疗抑郁症的用途和药物组合物 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810432839.0A Division CN108853505B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道抑制剂治疗抑郁症的用途和药物组合物 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115006534A CN115006534A (zh) | 2022-09-06 |
CN115006534B true CN115006534B (zh) | 2023-11-21 |
Family
ID=64104328
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210060456.1A Active CN115006534B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道Kir4.1抑制剂治疗抑郁症的用途和药物组合物 |
CN201810432839.0A Active CN108853505B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道抑制剂治疗抑郁症的用途和药物组合物 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810432839.0A Active CN108853505B (zh) | 2017-05-09 | 2018-05-08 | 钾离子通道抑制剂治疗抑郁症的用途和药物组合物 |
Country Status (4)
Country | Link |
---|---|
US (2) | US11326168B2 (zh) |
EP (1) | EP3622958B1 (zh) |
CN (2) | CN115006534B (zh) |
WO (1) | WO2018205927A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112717134B (zh) * | 2021-01-15 | 2022-05-20 | 山东大学齐鲁医院 | 一种用于儿童情感障碍的基因药物 |
CN118576596A (zh) * | 2023-03-01 | 2024-09-03 | 中国科学院上海药物研究所 | Lys01或其盐在制备Kir4.1钾离子通道抑制剂中的应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104338135A (zh) * | 2013-08-09 | 2015-02-11 | 中国科学院上海生命科学研究院 | 抑郁症的调节因子及其应用 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005054848A2 (en) * | 2003-11-25 | 2005-06-16 | Bayer Healthcare Ag | Diagnostics and therapeutics for diseases associated with g-protein-coupled inwardly rectifying potassium channel (girk2) |
CN101043891A (zh) * | 2004-09-18 | 2007-09-26 | 巴尔的摩马里兰大学 | 靶向NCCa-ATP通道的治疗剂及其使用方法 |
EP2530088A1 (en) * | 2011-05-30 | 2012-12-05 | Klinikum rechts der Isar der Technischen Universität München | Means and methods for diagnosing and treating multiple sclerosis |
EP2887806B1 (en) * | 2012-07-20 | 2019-11-13 | University Of Rochester | Method of treating and preventing brain impairment using na+-k+ -2ci- cotransporter isoform 1 inhibitors |
WO2018205935A1 (zh) * | 2017-05-09 | 2018-11-15 | 浙江大学 | 治疗抑郁症的方法和药物组合物 |
-
2018
- 2018-05-08 CN CN202210060456.1A patent/CN115006534B/zh active Active
- 2018-05-08 CN CN201810432839.0A patent/CN108853505B/zh active Active
- 2018-05-08 EP EP18798117.0A patent/EP3622958B1/en active Active
- 2018-05-08 WO PCT/CN2018/086021 patent/WO2018205927A1/zh unknown
-
2019
- 2019-11-09 US US16/679,197 patent/US11326168B2/en active Active
-
2022
- 2022-04-11 US US17/717,140 patent/US20230050684A1/en active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104338135A (zh) * | 2013-08-09 | 2015-02-11 | 中国科学院上海生命科学研究院 | 抑郁症的调节因子及其应用 |
Non-Patent Citations (3)
Title |
---|
"A disulphide-linked heterodimer of TWIK-1 and TREK-1 mediates passive conductance in astrocytes",Eun Mi Hwang等,《NATURE COMMUNICATIONS》,第1-15页;Eun Mi Hwang等;《NATURE COMMUNICATIONS》;20140205;第1-15页 * |
"Inhibition of astroglial Kir4.1 channels by selective serotonin reuptake inhibitors", Yukihiro Ohno等, 《Brain research》, 第1178卷, 第44-51页;Yukihiro Ohno等;《Brain research》;20070816;第1178卷;第44-51页 * |
Astroglial Kir4.1 in the lateral habenula drives neuronal bursts in depression;Yihui Cui等;《NATURE》;第323-342页 * |
Also Published As
Publication number | Publication date |
---|---|
EP3622958A4 (en) | 2021-02-24 |
WO2018205927A1 (zh) | 2018-11-15 |
EP3622958B1 (en) | 2024-03-20 |
CN108853505B (zh) | 2022-02-25 |
EP3622958A1 (en) | 2020-03-18 |
US20200149051A1 (en) | 2020-05-14 |
US20230050684A1 (en) | 2023-02-16 |
CN108853505A (zh) | 2018-11-23 |
US11326168B2 (en) | 2022-05-10 |
CN115006534A (zh) | 2022-09-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Gaudet et al. | miR-155 deletion in mice overcomes neuron-intrinsic and neuron-extrinsic barriers to spinal cord repair | |
Chauvin et al. | Neuronal stathmins: a family of phosphoproteins cooperating for neuronal development, plasticity and regeneration | |
Dell'Orco et al. | Neuronal atrophy early in degenerative ataxia is a compensatory mechanism to regulate membrane excitability | |
Tang et al. | Survival effect of PDGF-CC rescues neurons from apoptosis in both brain and retina by regulating GSK3β phosphorylation | |
Boido et al. | Increasing agrin function antagonizes muscle atrophy and motor impairment in spinal muscular atrophy | |
JP6755938B2 (ja) | アルツハイマー病の治療におけるPI4KIIIαタンパク質および関連膜タンパク質複合体の利用 | |
US9486521B2 (en) | Therapeutic applications targeting SARM1 | |
Shen et al. | Role of Fto on CaMKII/CREB signaling pathway of hippocampus in depressive-like behaviors induced by chronic restraint stress mice | |
Li et al. | MicroRNA-125b mimic inhibits ischemia reperfusion-induced neuroinflammation and aberrant p53 apoptotic signalling activation through targeting TP53INP1 | |
Kusakari et al. | Shp2 in forebrain neurons regulates synaptic plasticity, locomotion, and memory formation in mice | |
US20230050684A1 (en) | Use of potassium channel inhibitor for treating depression | |
Han et al. | Lentiviral-mediated netrin-1 overexpression improves motor and sensory functions in SCT rats associated with SYP and GAP-43 expressions | |
WO2017201425A1 (en) | Anabolic enhancers for ameliorating neurodegeneration | |
KR102488987B1 (ko) | 알츠하이머 질환을 치료하는데 사용되는 약물 및 치료 표적제를 스크리닝하는 방법 | |
Wei et al. | Strain-specific BDNF expression of rat primary astrocytes | |
Zhou et al. | CircDYM attenuates microglial apoptosis via CEBPB/ZC3H4 axis in LPS-induced mouse model of depression | |
Boutary | Development of a Targeted Therapy for Charcot Marie Tooth 1A (CMT1A) Neuropathy Based on siRNA PMP22 Squalene Nanoparticles | |
Sun et al. | The MuSK agonist antibody protects the neuromuscular junction and extends the lifespan in C9orf72-ALS mice | |
US20160074467A1 (en) | Treatment of aging effects by gonadotropin-releasing hormone, neurogenesis or brain ikk beta/nf-kappab inhibition | |
Tassinari | Development of an innovative strategy to enhance the efficacy of gene therapy for CDKL5 deficiency disorder | |
Loan et al. | Treatment options in motor neuron disease: Amyotrophic lateral sclerosis and spinal muscular atrophy | |
Zhen | Protocadherin 15 modulates oligodendrocyte progenitor cell proliferation and contact-mediated self-repulsion through two distinct signalling pathways | |
Szallasi | Targeting TRP channels for anxiety relief and improving mental health | |
Eneanya | MURBA-22 3p and Its Role in Tau Phosphorylation | |
Kharouf et al. | The hyperpolarization-activated cyclic nucleotide-gated 4 channel as a potential anti-seizure |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |