JP6039705B2 - タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 - Google Patents
タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 Download PDFInfo
- Publication number
- JP6039705B2 JP6039705B2 JP2015003310A JP2015003310A JP6039705B2 JP 6039705 B2 JP6039705 B2 JP 6039705B2 JP 2015003310 A JP2015003310 A JP 2015003310A JP 2015003310 A JP2015003310 A JP 2015003310A JP 6039705 B2 JP6039705 B2 JP 6039705B2
- Authority
- JP
- Japan
- Prior art keywords
- tau
- fkbp52
- protein
- polypeptide
- cells
- 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
- 230000004064 dysfunction Effects 0.000 title claims description 37
- 208000012902 Nervous system disease Diseases 0.000 title claims description 22
- 230000003993 interaction Effects 0.000 title description 46
- 208000025966 Neurological disease Diseases 0.000 title description 11
- 230000001225 therapeutic effect Effects 0.000 title description 6
- 108010026424 tau Proteins Proteins 0.000 claims description 267
- 102000013498 tau Proteins Human genes 0.000 claims description 267
- 102100020739 Peptidyl-prolyl cis-trans isomerase FKBP4 Human genes 0.000 claims description 236
- 108010067247 tacrolimus binding protein 4 Proteins 0.000 claims description 235
- 238000000034 method Methods 0.000 claims description 91
- 108090000623 proteins and genes Proteins 0.000 claims description 73
- 230000027455 binding Effects 0.000 claims description 57
- 102000004169 proteins and genes Human genes 0.000 claims description 57
- 208000024827 Alzheimer disease Diseases 0.000 claims description 24
- 201000001119 neuropathy Diseases 0.000 claims description 16
- 230000007823 neuropathy Effects 0.000 claims description 16
- 208000033808 peripheral neuropathy Diseases 0.000 claims description 16
- 108091023037 Aptamer Proteins 0.000 claims description 11
- 230000008569 process Effects 0.000 claims description 6
- 210000002966 serum Anatomy 0.000 claims description 5
- 238000010998 test method Methods 0.000 claims description 3
- 238000001574 biopsy Methods 0.000 claims description 2
- 210000004369 blood Anatomy 0.000 claims description 2
- 239000008280 blood Substances 0.000 claims description 2
- 210000001175 cerebrospinal fluid Anatomy 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 210000002700 urine Anatomy 0.000 claims description 2
- 210000005013 brain tissue Anatomy 0.000 claims 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 119
- 102000004196 processed proteins & peptides Human genes 0.000 description 115
- 229920001184 polypeptide Polymers 0.000 description 109
- 210000004027 cell Anatomy 0.000 description 77
- 150000001875 compounds Chemical class 0.000 description 74
- 235000018102 proteins Nutrition 0.000 description 52
- 239000000523 sample Substances 0.000 description 43
- 230000014509 gene expression Effects 0.000 description 39
- 238000003556 assay Methods 0.000 description 36
- 230000000694 effects Effects 0.000 description 32
- 238000012216 screening Methods 0.000 description 30
- 238000009825 accumulation Methods 0.000 description 27
- 210000004556 brain Anatomy 0.000 description 26
- 108010070675 Glutathione transferase Proteins 0.000 description 23
- 102100029100 Hematopoietic prostaglandin D synthase Human genes 0.000 description 23
- 239000000427 antigen Substances 0.000 description 22
- 108091007433 antigens Proteins 0.000 description 21
- 102000036639 antigens Human genes 0.000 description 21
- 102000029749 Microtubule Human genes 0.000 description 20
- 108091022875 Microtubule Proteins 0.000 description 20
- 108091023040 Transcription factor Proteins 0.000 description 19
- 102000040945 Transcription factor Human genes 0.000 description 19
- 210000004688 microtubule Anatomy 0.000 description 19
- 238000012360 testing method Methods 0.000 description 19
- 230000002776 aggregation Effects 0.000 description 18
- 238000004220 aggregation Methods 0.000 description 18
- 108020004414 DNA Proteins 0.000 description 17
- 230000003915 cell function Effects 0.000 description 17
- 230000006870 function Effects 0.000 description 17
- 230000004048 modification Effects 0.000 description 17
- 238000012986 modification Methods 0.000 description 17
- 238000011282 treatment Methods 0.000 description 17
- 238000001262 western blot Methods 0.000 description 17
- 230000001404 mediated effect Effects 0.000 description 16
- 108020004999 messenger RNA Proteins 0.000 description 16
- 230000001323 posttranslational effect Effects 0.000 description 16
- 102100023995 Beta-nerve growth factor Human genes 0.000 description 15
- 108010025020 Nerve Growth Factor Proteins 0.000 description 15
- 229940053128 nerve growth factor Drugs 0.000 description 15
- 150000007523 nucleic acids Chemical group 0.000 description 15
- 102000009658 Peptidylprolyl Isomerase Human genes 0.000 description 14
- 108010020062 Peptidylprolyl Isomerase Proteins 0.000 description 14
- 102000001708 Protein Isoforms Human genes 0.000 description 14
- 108010029485 Protein Isoforms Proteins 0.000 description 14
- 239000000758 substrate Substances 0.000 description 14
- 241000700159 Rattus Species 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 12
- 239000003446 ligand Substances 0.000 description 12
- 102000039446 nucleic acids Human genes 0.000 description 12
- 108020004707 nucleic acids Proteins 0.000 description 12
- 230000026731 phosphorylation Effects 0.000 description 12
- 238000006366 phosphorylation reaction Methods 0.000 description 12
- 230000014511 neuron projection development Effects 0.000 description 11
- 229920002704 polyhistidine Polymers 0.000 description 11
- 108091028043 Nucleic acid sequence Proteins 0.000 description 10
- 125000003275 alpha amino acid group Chemical group 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 10
- 239000000872 buffer Substances 0.000 description 10
- 239000012634 fragment Substances 0.000 description 10
- MQLACMBJVPINKE-UHFFFAOYSA-N 10-[(3-hydroxy-4-methoxyphenyl)methylidene]anthracen-9-one Chemical compound C1=C(O)C(OC)=CC=C1C=C1C2=CC=CC=C2C(=O)C2=CC=CC=C21 MQLACMBJVPINKE-UHFFFAOYSA-N 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- 210000002241 neurite Anatomy 0.000 description 9
- 230000002265 prevention Effects 0.000 description 9
- 230000004568 DNA-binding Effects 0.000 description 8
- 201000011240 Frontotemporal dementia Diseases 0.000 description 8
- 108010004469 allophycocyanin Proteins 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 239000000284 extract Substances 0.000 description 8
- 239000000499 gel Substances 0.000 description 8
- 238000003364 immunohistochemistry Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 108010001515 Galectin 4 Proteins 0.000 description 7
- 102100039556 Galectin-4 Human genes 0.000 description 7
- 108091030071 RNAI Proteins 0.000 description 7
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 7
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 7
- 108090000704 Tubulin Proteins 0.000 description 7
- 102000004243 Tubulin Human genes 0.000 description 7
- 239000012190 activator Substances 0.000 description 7
- 230000001086 cytosolic effect Effects 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 229940088598 enzyme Drugs 0.000 description 7
- 230000009368 gene silencing by RNA Effects 0.000 description 7
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 7
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 7
- 230000001506 immunosuppresive effect Effects 0.000 description 7
- 239000012528 membrane Substances 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- 239000008188 pellet Substances 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 7
- 238000003127 radioimmunoassay Methods 0.000 description 7
- 230000001105 regulatory effect Effects 0.000 description 7
- 239000013598 vector Substances 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 101710137189 Amyloid-beta A4 protein Proteins 0.000 description 6
- 102100022704 Amyloid-beta precursor protein Human genes 0.000 description 6
- 101710151993 Amyloid-beta precursor protein Proteins 0.000 description 6
- 102000053642 Catalytic RNA Human genes 0.000 description 6
- 108090000994 Catalytic RNA Proteins 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- DZHSAHHDTRWUTF-SIQRNXPUSA-N amyloid-beta polypeptide 42 Chemical compound C([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O)[C@@H](C)CC)C(C)C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C(C)C)C1=CC=CC=C1 DZHSAHHDTRWUTF-SIQRNXPUSA-N 0.000 description 6
- 239000011324 bead Substances 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000004113 cell culture Methods 0.000 description 6
- 238000002868 homogeneous time resolved fluorescence Methods 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 238000003199 nucleic acid amplification method Methods 0.000 description 6
- 230000002018 overexpression Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 230000002285 radioactive effect Effects 0.000 description 6
- 108091092562 ribozyme Proteins 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 241000588724 Escherichia coli Species 0.000 description 5
- 101100402621 Homo sapiens MSANTD4 gene Proteins 0.000 description 5
- 102000009664 Microtubule-Associated Proteins Human genes 0.000 description 5
- 108010020004 Microtubule-Associated Proteins Proteins 0.000 description 5
- 102100031642 Myb/SANT-like DNA-binding domain-containing protein 4 Human genes 0.000 description 5
- 239000000020 Nitrocellulose Substances 0.000 description 5
- 101710147152 Peptidyl-prolyl cis-trans isomerase FKBP4 Proteins 0.000 description 5
- 230000003321 amplification Effects 0.000 description 5
- 230000008045 co-localization Effects 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 210000003618 cortical neuron Anatomy 0.000 description 5
- 210000000172 cytosol Anatomy 0.000 description 5
- 230000007423 decrease Effects 0.000 description 5
- 208000035475 disorder Diseases 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 238000003119 immunoblot Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 229920001220 nitrocellulos Polymers 0.000 description 5
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 5
- 230000014616 translation Effects 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 4
- 101710113864 Heat shock protein 90 Proteins 0.000 description 4
- 102100034051 Heat shock protein HSP 90-alpha Human genes 0.000 description 4
- 108010016648 Immunophilins Proteins 0.000 description 4
- 102000000521 Immunophilins Human genes 0.000 description 4
- 108010027179 Tacrolimus Binding Proteins Proteins 0.000 description 4
- 102000018679 Tacrolimus Binding Proteins Human genes 0.000 description 4
- 238000013459 approach Methods 0.000 description 4
- 210000003050 axon Anatomy 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 238000003018 immunoassay Methods 0.000 description 4
- 238000002372 labelling Methods 0.000 description 4
- 235000013336 milk Nutrition 0.000 description 4
- 239000008267 milk Substances 0.000 description 4
- 210000004080 milk Anatomy 0.000 description 4
- 210000002569 neuron Anatomy 0.000 description 4
- 238000006384 oligomerization reaction Methods 0.000 description 4
- 230000004481 post-translational protein modification Effects 0.000 description 4
- 230000004850 protein–protein interaction Effects 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 238000004062 sedimentation Methods 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 238000013518 transcription Methods 0.000 description 4
- 230000035897 transcription Effects 0.000 description 4
- 238000013519 translation Methods 0.000 description 4
- SGKRLCUYIXIAHR-AKNGSSGZSA-N (4s,4ar,5s,5ar,6r,12ar)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4a,5,5a,6-tetrahydro-4h-tetracene-2-carboxamide Chemical compound C1=CC=C2[C@H](C)[C@@H]([C@H](O)[C@@H]3[C@](C(O)=C(C(N)=O)C(=O)[C@H]3N(C)C)(O)C3=O)C3=C(O)C2=C1O SGKRLCUYIXIAHR-AKNGSSGZSA-N 0.000 description 3
- 102100027831 14-3-3 protein theta Human genes 0.000 description 3
- 108020004491 Antisense DNA Proteins 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 3
- 101000891579 Homo sapiens Microtubule-associated protein tau Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 108091034117 Oligonucleotide Proteins 0.000 description 3
- 208000000609 Pick Disease of the Brain Diseases 0.000 description 3
- 229920002684 Sepharose Polymers 0.000 description 3
- 108010085012 Steroid Receptors Proteins 0.000 description 3
- 102000007451 Steroid Receptors Human genes 0.000 description 3
- 239000007983 Tris buffer Substances 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000004075 alteration Effects 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 239000003816 antisense DNA Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 229960003722 doxycycline Drugs 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 108020001507 fusion proteins Proteins 0.000 description 3
- 102000037865 fusion proteins Human genes 0.000 description 3
- 238000001502 gel electrophoresis Methods 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 3
- 102000057063 human MAPT Human genes 0.000 description 3
- 230000006951 hyperphosphorylation Effects 0.000 description 3
- 229940127121 immunoconjugate Drugs 0.000 description 3
- 238000001114 immunoprecipitation Methods 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 210000004962 mammalian cell Anatomy 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 230000004770 neurodegeneration Effects 0.000 description 3
- 208000015122 neurodegenerative disease Diseases 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000001590 oxidative effect Effects 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 238000006116 polymerization reaction Methods 0.000 description 3
- 210000004129 prosencephalon Anatomy 0.000 description 3
- 238000010379 pull-down assay Methods 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 230000000717 retained effect Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 108700028369 Alleles Proteins 0.000 description 2
- 108020005544 Antisense RNA Proteins 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 102100026189 Beta-galactosidase Human genes 0.000 description 2
- 208000014644 Brain disease Diseases 0.000 description 2
- 101000746263 Conus leopardus Conotoxin Lp5.1 Proteins 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 101710103508 FK506-binding protein Proteins 0.000 description 2
- 101710104425 FK506-binding protein 2 Proteins 0.000 description 2
- 101710104423 FK506-binding protein 3 Proteins 0.000 description 2
- 101710104333 FK506-binding protein 4 Proteins 0.000 description 2
- 101710104342 FK506-binding protein 5 Proteins 0.000 description 2
- 101710149710 FKBP-type 16 kDa peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 101710121306 FKBP-type 22 kDa peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 101710180800 FKBP-type peptidyl-prolyl cis-trans isomerase FkpA Proteins 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 101000979001 Homo sapiens Methionine aminopeptidase 2 Proteins 0.000 description 2
- 101000969087 Homo sapiens Microtubule-associated protein 2 Proteins 0.000 description 2
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 description 2
- 101710104030 Long-type peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102100023174 Methionine aminopeptidase 2 Human genes 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 102000005431 Molecular Chaperones Human genes 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 2
- 108010038807 Oligopeptides Proteins 0.000 description 2
- 102000015636 Oligopeptides Human genes 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 101710114693 Outer membrane protein MIP Proteins 0.000 description 2
- 101001128814 Pandinus imperator Pandinin-1 Proteins 0.000 description 2
- 208000018737 Parkinson disease Diseases 0.000 description 2
- 101710116692 Peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 101710111764 Peptidyl-prolyl cis-trans isomerase FKBP10 Proteins 0.000 description 2
- 101710111749 Peptidyl-prolyl cis-trans isomerase FKBP11 Proteins 0.000 description 2
- 101710111747 Peptidyl-prolyl cis-trans isomerase FKBP12 Proteins 0.000 description 2
- 101710111757 Peptidyl-prolyl cis-trans isomerase FKBP14 Proteins 0.000 description 2
- 101710111682 Peptidyl-prolyl cis-trans isomerase FKBP1A Proteins 0.000 description 2
- 101710111689 Peptidyl-prolyl cis-trans isomerase FKBP1B Proteins 0.000 description 2
- 101710147154 Peptidyl-prolyl cis-trans isomerase FKBP2 Proteins 0.000 description 2
- 101710147149 Peptidyl-prolyl cis-trans isomerase FKBP3 Proteins 0.000 description 2
- 101710147150 Peptidyl-prolyl cis-trans isomerase FKBP5 Proteins 0.000 description 2
- 101710147138 Peptidyl-prolyl cis-trans isomerase FKBP7 Proteins 0.000 description 2
- 101710147137 Peptidyl-prolyl cis-trans isomerase FKBP8 Proteins 0.000 description 2
- 101710147136 Peptidyl-prolyl cis-trans isomerase FKBP9 Proteins 0.000 description 2
- 101710174853 Peptidyl-prolyl cis-trans isomerase Mip Proteins 0.000 description 2
- 101710200991 Peptidyl-prolyl cis-trans isomerase, rhodopsin-specific isozyme Proteins 0.000 description 2
- 101710092145 Peptidyl-prolyl cis-trans isomerase-like 1 Proteins 0.000 description 2
- 101710092146 Peptidyl-prolyl cis-trans isomerase-like 2 Proteins 0.000 description 2
- 101710092148 Peptidyl-prolyl cis-trans isomerase-like 3 Proteins 0.000 description 2
- 101710092149 Peptidyl-prolyl cis-trans isomerase-like 4 Proteins 0.000 description 2
- 108010004729 Phycoerythrin Proteins 0.000 description 2
- 101710113444 Probable parvulin-type peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 101710090737 Probable peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 101710133309 Putative peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- 101710124237 Short-type peptidyl-prolyl cis-trans isomerase Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 208000034799 Tauopathies Diseases 0.000 description 2
- 239000004098 Tetracycline Substances 0.000 description 2
- 108090000190 Thrombin Proteins 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000021736 acetylation Effects 0.000 description 2
- 238000006640 acetylation reaction Methods 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- 210000004227 basal ganglia Anatomy 0.000 description 2
- 108010005774 beta-Galactosidase Proteins 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000002759 chromosomal effect Effects 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 238000011490 co-immunoprecipitation assay Methods 0.000 description 2
- 208000010877 cognitive disease Diseases 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 239000003184 complementary RNA Substances 0.000 description 2
- 238000003271 compound fluorescence assay Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 239000006059 cover glass Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 238000012137 double-staining Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 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 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- GWQVMPWSEVRGPY-UHFFFAOYSA-N europium cryptate Chemical compound [Eu+3].N=1C2=CC=CC=1CN(CC=1N=C(C=CC=1)C=1N=C(C3)C=CC=1)CC(N=1)=CC(C(=O)NCCN)=CC=1C(N=1)=CC(C(=O)NCCN)=CC=1CN3CC1=CC=CC2=N1 GWQVMPWSEVRGPY-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 210000000020 growth cone Anatomy 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000002452 interceptive effect Effects 0.000 description 2
- 229910052747 lanthanoid Inorganic materials 0.000 description 2
- 150000002602 lanthanoids Chemical class 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 230000001537 neural effect Effects 0.000 description 2
- 230000000324 neuroprotective effect Effects 0.000 description 2
- 238000010606 normalization Methods 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 229960003330 pentetic acid Drugs 0.000 description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 239000003504 photosensitizing agent Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 201000002212 progressive supranuclear palsy Diseases 0.000 description 2
- 238000000159 protein binding assay Methods 0.000 description 2
- 230000004853 protein function Effects 0.000 description 2
- LISFMEBWQUVKPJ-UHFFFAOYSA-N quinolin-2-ol Chemical compound C1=CC=C2NC(=O)C=CC2=C1 LISFMEBWQUVKPJ-UHFFFAOYSA-N 0.000 description 2
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 229960002930 sirolimus Drugs 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 229960002180 tetracycline Drugs 0.000 description 2
- 229930101283 tetracycline Natural products 0.000 description 2
- 235000019364 tetracycline Nutrition 0.000 description 2
- 150000003522 tetracyclines Chemical class 0.000 description 2
- 229960004072 thrombin Drugs 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000003160 two-hybrid assay Methods 0.000 description 2
- KWPACVJPAFGBEQ-IKGGRYGDSA-N (2s)-1-[(2r)-2-amino-3-phenylpropanoyl]-n-[(3s)-1-chloro-6-(diaminomethylideneamino)-2-oxohexan-3-yl]pyrrolidine-2-carboxamide Chemical compound C([C@@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)CCl)C1=CC=CC=C1 KWPACVJPAFGBEQ-IKGGRYGDSA-N 0.000 description 1
- CZIHNRWJTSTCEX-UHFFFAOYSA-N 2 Acetylaminofluorene Chemical compound C1=CC=C2C3=CC=C(NC(=O)C)C=C3CC2=C1 CZIHNRWJTSTCEX-UHFFFAOYSA-N 0.000 description 1
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 description 1
- JFJNVIPVOCESGZ-UHFFFAOYSA-N 2,3-dipyridin-2-ylpyridine Chemical compound N1=CC=CC=C1C1=CC=CN=C1C1=CC=CC=N1 JFJNVIPVOCESGZ-UHFFFAOYSA-N 0.000 description 1
- SUBDBMMJDZJVOS-UHFFFAOYSA-N 5-methoxy-2-{[(4-methoxy-3,5-dimethylpyridin-2-yl)methyl]sulfinyl}-1H-benzimidazole Chemical compound N=1C2=CC(OC)=CC=C2NC=1S(=O)CC1=NC=C(C)C(OC)=C1C SUBDBMMJDZJVOS-UHFFFAOYSA-N 0.000 description 1
- 108091027075 5S-rRNA precursor Proteins 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 239000012099 Alexa Fluor family Substances 0.000 description 1
- 208000000044 Amnesia Diseases 0.000 description 1
- 208000031091 Amnestic disease Diseases 0.000 description 1
- 108020000948 Antisense Oligonucleotides Proteins 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 230000006974 Aβ toxicity Effects 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- ROFVEXUMMXZLPA-UHFFFAOYSA-N Bipyridyl Chemical compound N1=CC=CC=C1C1=CC=CC=N1 ROFVEXUMMXZLPA-UHFFFAOYSA-N 0.000 description 1
- QYOVMAREBTZLBT-KTKRTIGZSA-N CCCCCCCC\C=C/CCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO Chemical compound CCCCCCCC\C=C/CCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO QYOVMAREBTZLBT-KTKRTIGZSA-N 0.000 description 1
- 102000004631 Calcineurin Human genes 0.000 description 1
- 108010042955 Calcineurin Proteins 0.000 description 1
- 102000000584 Calmodulin Human genes 0.000 description 1
- 108010041952 Calmodulin Proteins 0.000 description 1
- 102000052052 Casein Kinase II Human genes 0.000 description 1
- 108010010919 Casein Kinase II Proteins 0.000 description 1
- 206010010099 Combined immunodeficiency Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 1
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 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
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 1
- 229910052693 Europium Inorganic materials 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 208000002339 Frontotemporal Lobar Degeneration Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 101710115937 Microtubule-associated protein tau Proteins 0.000 description 1
- 102100040243 Microtubule-associated protein tau Human genes 0.000 description 1
- 108010085220 Multiprotein Complexes Proteins 0.000 description 1
- 102000007474 Multiprotein Complexes Human genes 0.000 description 1
- 101100334858 Mus musculus Fkbp4 gene Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 108091007491 NSP3 Papain-like protease domains Proteins 0.000 description 1
- 241000244206 Nematoda Species 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 1
- 239000007990 PIPES buffer Substances 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 208000027089 Parkinsonian disease Diseases 0.000 description 1
- 206010034010 Parkinsonism Diseases 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108010079855 Peptide Aptamers Proteins 0.000 description 1
- 102100027913 Peptidyl-prolyl cis-trans isomerase FKBP1A Human genes 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 108010066717 Q beta Replicase Proteins 0.000 description 1
- 108090000244 Rat Proteins Proteins 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 108091027981 Response element Proteins 0.000 description 1
- 241000206572 Rhodophyta Species 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- 241000235347 Schizosaccharomyces pombe Species 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 238000010161 Student-Newman-Keuls test Methods 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 108010006877 Tacrolimus Binding Protein 1A Proteins 0.000 description 1
- 102100036407 Thioredoxin Human genes 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 230000006986 amnesia Effects 0.000 description 1
- 238000004873 anchoring Methods 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 239000000074 antisense oligonucleotide Substances 0.000 description 1
- 238000012230 antisense oligonucleotides Methods 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- 230000008335 axon cargo transport Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000004067 bulking agent Substances 0.000 description 1
- 210000002230 centromere Anatomy 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000012412 chemical coupling Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000012504 chromatography matrix Substances 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 208000013044 corticobasal degeneration disease Diseases 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 description 1
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- SLPJGDQJLTYWCI-UHFFFAOYSA-N dimethyl-(4,5,6,7-tetrabromo-1h-benzoimidazol-2-yl)-amine Chemical compound BrC1=C(Br)C(Br)=C2NC(N(C)C)=NC2=C1Br SLPJGDQJLTYWCI-UHFFFAOYSA-N 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 230000002616 endonucleolytic effect Effects 0.000 description 1
- 230000006862 enzymatic digestion Effects 0.000 description 1
- OGPBJKLSAFTDLK-UHFFFAOYSA-N europium atom Chemical compound [Eu] OGPBJKLSAFTDLK-UHFFFAOYSA-N 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 102000034287 fluorescent proteins Human genes 0.000 description 1
- 108091006047 fluorescent proteins Proteins 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 210000005104 human peripheral blood lymphocyte Anatomy 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 238000000760 immunoelectrophoresis Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 239000012133 immunoprecipitate Substances 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 230000001861 immunosuppressant effect Effects 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000009878 intermolecular interaction Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 102000027411 intracellular receptors Human genes 0.000 description 1
- 108091008582 intracellular receptors Proteins 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 239000013038 irreversible inhibitor Substances 0.000 description 1
- 238000003367 kinetic assay Methods 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000007834 ligase chain reaction Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- UNSKIISCJFJWTL-WIDVRGBZSA-N meridamycin Chemical compound C1C(O)C(C)C(O)CC(O)CC(O)\C(C)=C\C(C)CC(C)C(O)C(/CC)=C/CC(C(\C)=C\C(C)C(C)O)OC(=O)C2CCCN2C(=O)C(=O)C2(O)C(C)CCC1O2 UNSKIISCJFJWTL-WIDVRGBZSA-N 0.000 description 1
- DGKUOWHAUIWQTM-UHFFFAOYSA-N meridamycin Natural products C1C(O)C(C)C(O)CC(O)CC(O)C(C)=CC(C)CC(C)C(O)C(CC)=CCC(C(C)=CC(C)C(C)O)OC(=O)C2CCCCN2C(=O)C(=O)C2(O)C(C)CCC1O2 DGKUOWHAUIWQTM-UHFFFAOYSA-N 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 238000010232 migration assay Methods 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 230000037023 motor activity Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 210000002682 neurofibrillary tangle Anatomy 0.000 description 1
- 230000004112 neuroprotection Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- QNDVLZJODHBUFM-WFXQOWMNSA-N okadaic acid Chemical compound C([C@H](O1)[C@H](C)/C=C/[C@H]2CC[C@@]3(CC[C@H]4O[C@@H](C([C@@H](O)[C@@H]4O3)=C)[C@@H](O)C[C@H](C)[C@@H]3[C@@H](CC[C@@]4(OCCCC4)O3)C)O2)C(C)=C[C@]21O[C@H](C[C@@](C)(O)C(O)=O)CC[C@H]2O QNDVLZJODHBUFM-WFXQOWMNSA-N 0.000 description 1
- VEFJHAYOIAAXEU-UHFFFAOYSA-N okadaic acid Natural products CC(CC(O)C1OC2CCC3(CCC(O3)C=CC(C)C4CC(=CC5(OC(CC(C)(O)C(=O)O)CCC5O)O4)C)OC2C(O)C1C)C6OC7(CCCCO7)CCC6C VEFJHAYOIAAXEU-UHFFFAOYSA-N 0.000 description 1
- 229940124276 oligodeoxyribonucleotide Drugs 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229960003104 ornithine Drugs 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 231100000255 pathogenic effect Toxicity 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000816 peptidomimetic Substances 0.000 description 1
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229920000447 polyanionic polymer Polymers 0.000 description 1
- 229920005646 polycarboxylate Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 229920000915 polyvinyl chloride Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 108020001580 protein domains Proteins 0.000 description 1
- 230000006916 protein interaction Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 239000012857 radioactive material Substances 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 239000012217 radiopharmaceutical Substances 0.000 description 1
- 229940121896 radiopharmaceutical Drugs 0.000 description 1
- 230000002799 radiopharmaceutical effect Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 230000020273 regulation of microtubule cytoskeleton organization Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000002629 repopulating effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007894 restriction fragment length polymorphism technique Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000009919 sequestration Effects 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 208000019116 sleep disease Diseases 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 238000012453 sprague-dawley rat model Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000004960 subcellular localization Effects 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 108060008226 thioredoxin Proteins 0.000 description 1
- 229940094937 thioredoxin Drugs 0.000 description 1
- 238000002877 time resolved fluorescence resonance energy transfer Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000009966 trimming Methods 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 238000010396 two-hybrid screening Methods 0.000 description 1
- 108020005087 unfolded proteins Proteins 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
Images
Classifications
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4709—Amyloid plaque core protein
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/99—Isomerases (5.)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/02—Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2814—Dementia; Cognitive disorders
- G01N2800/2821—Alzheimer
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2814—Dementia; Cognitive disorders
- G01N2800/2828—Prion diseases
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2835—Movement disorders, e.g. Parkinson, Huntington, Tourette
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Epidemiology (AREA)
- Hospice & Palliative Care (AREA)
- Psychiatry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
本発明は、全般に、タウ機能不全を伴う神経障害(アルツハイマー病が含まれる)における神経保護および修復に関する。本発明は、タンパク質FKBP52とタウとの間の直接相互作用を記載する。本発明は、病原性タウの有害作用をモデュレーションするための、FKBP52−タウ相互作用に作用する分子についてのスクリーニング法に関する。本発明は、最終的には、タウ機能不全を伴う神経障害の治療的診断アッセイ、予後アッセイ、およびモニタリングアッセイに関する。
タウタンパク質は、中枢神経系、主にニューロンに広く発現され、そこで微小管動態、軸索輸送および神経突起伸長の調節に重要な役割を果たす主要な微小管関連タンパク質(MAP)である。タンパク質タウは、17番染色体上に局在する独自の遺伝子の一次転写物のエクソン2、3および10の選択的スプライシングによって生成する6種の異なるアイソフォームで成体ヒト脳に存在する。それらの配列長は、352から441個のアミノ酸まで変動する。ますます増える証拠から、「凝集した」タウ(ネイティブな未フォールディングのタンパク質)の異常集合が、アルツハイマー病、ピック病、脳皮質基底核変性症、軽度認知機能不全、進行性核上性麻痺、および17番染色体に連鎖しパーキンソニズムを伴う前頭側頭型認知症が含まれる、タウオパチーまたはタウ機能不全を伴う神経障害とまとめて呼ばれる一連のヒト認知疾患の特徴であることが示唆される。過剰リン酸化、突然変異、短縮化および「濃縮体(tangle)」への凝集などのタウの異常が、病原過程の要因でありうる。今日まで、神経変性疾患の誘導にタウ改変が果たす役割は、完全には理解されていないことから、タウの構造/機能をコントロールする分子メカニズムを解釈することは、大いなる関心対象であり、これらの疾患のための新規な治療アプローチを見出すことを助ける可能性がある。
本発明者らは、イムノフィリンFKBP52と微小管関連タンパク質タウ(過剰リン酸化されているか、またはされていないその公知の全てのアイソフォームに属する)との間に直接的で特異的なタンパク質−タンパク質相互作用を発見した。本発明は、FKBP52−タウ相互作用が、タウ機能不全を伴う神経障害の新規な治療アプローチのために、特にアルツハイマー病のために好都合に使用されうる新しいターゲットを提供することを立証する。
本発明のスクリーニング法:
本発明の第一の目的は、タウ機能不全を伴う神経障害の予防および治療のための薬物をスクリーニングするための方法であって、以下のステップを含む方法から成る:
a)候補化合物がタウポリペプチドとFKBP52ポリペプチドとの間の相互作用をモデュレーションする能力を決定すること、および
b)該相互作用をモデュレーションする候補化合物を正に選択すること。
− a1)被験候補化合物を、第一タウポリペプチドまたはその実質的に相同もしくは実質的に類似のアミノ酸配列と、(2)第二FKBP52ポリペプチドまたはその実質的に相同もしくは実質的に類似のアミノ酸配列との混合物と接触させること、および
− a2)該候補化合物が、該タウポリペプチドと該第二FKBP52ポリペプチドとの間の結合をモデュレーションする能力を決定すること。
(1)
− (i)上記のタウポリペプチドと(ii)転写因子の第一タンパク質部分との間の第一融合ポリペプチド
− (i)上記のFKBP52ポリペプチドと(ii)転写因子の第二部分との間の第二融合ポリペプチド
を発現しているホスト細胞を提供すること
ここで、第一および第二タンパク質部分が一緒に結合している場合、該転写因子は、DNAのターゲット調節配列に対して活性であり、
該ホスト細胞は、(i)該活性転写因子によって活性化されうる調節DNA配列および(ii)該調節配列に作動連結されているDNAレポート配列を含む核酸も含有する、
(2)ステップ1)で提供された該ホスト細胞を被験候補化合物と接触させること、
(3)該DNAレポーター配列の発現レベルを決定すること。
(1)上記の第一タウポリペプチドおよび第二FKBP52ポリペプチドを提供すること、
(2)被験候補化合物を該ポリペプチドと接触させること、
(3)ステップ(2)で得られたような該ポリペプチドおよび該候補化合物を用いて適切な泳動基材上でゲル泳動アッセイを行うこと、
(4)ステップ(3)を行われた泳動アッセイで該ポリペプチドの間に形成した複合体を検出および定量すること。
(1)
− 第一抗原と融合された第一タウポリペプチド、
− 第二抗原と融合された第二FKBP52ポリペプチド、
− 被験候補化合物
を含むアッセイ前試料を接触させること、
(2)ステップ(2)の該アッセイ前試料に:
− 特異的に該第一抗原に対する、ユーロピアン(European)クリプテートでラベルされた少なくとも一つの抗体、
− 該第二抗原に対する、XL665でラベルされた少なくとも一つの抗体
を添加すること、
(3)ステップ(2)のアッセイ試料を該ユーロピアンクリプテートの励起波長で照射すること、
(4)XL665の発光波長で発光された蛍光シグナルを検出および/または定量すること、
(5)ステップ(4)で得られた蛍光シグナルを、ステップ(1)のアッセイ前試料が被験候補化合物の不在下で調製されたときに得られた蛍光と比較すること。
本明細書前記のin vitroスクリーニングの任意の一態様の終わりに正に選択された候補化合物は、タウ介在性細胞機能(チューブリン重合など)、タウ蓄積、タウ凝集および全ての翻訳後タウ改変に関するその性質をさらにアッセイすることを考慮してさらなる選択ステップに供することができる。このために、上記の全般的in vitroスクリーニング法を用いて正に選択された候補化合物は、さらに、それらがタウ介在性細胞機能、タウ蓄積、タウ凝集および全ての翻訳後タウ改変をモデュレーションする能力について選択することができる。
i)請求項1〜6のいずれか記載のin vitroスクリーニング法を行うことによって、タウとFKBP52タンパク質との間の相互作用をモデュレーションする化合物についてスクリーニングすること、および
ii)ステップi)の終わりに正に選択された化合物を、それらのタウ介在性細胞機能、タウ蓄積、タウ凝集および全ての翻訳後タウ改変についてスクリーニングすること。
(1)細胞を、ステップi)の終わりに正に選択された化合物と接触させること、
(2)化合物がタウ介在性細胞機能、タウ蓄積、タウ凝集および全ての翻訳後タウ改変をモデュレーションする潜在能力を決定すること、
(3)ステップ(2)で決定されたタウ介在性細胞機能、タウ蓄積、タウ凝集および全ての翻訳後タウ改変を、ステップ(1)が前記の正に選択された化合物の不在下で行われた場合に決定されたタウ介在性細胞機能、タウ蓄積、タウ凝集および全ての翻訳後タウ改変と比較すること。
本発明の一態様によると、候補化合物は、ペプチド、ペプチド模倣体、有機小分子、抗体、アプタマーまたは核酸から成る群より選択されることがある。例えば、本発明による候補化合物は、予め合成された化合物のライブラリー、または構造がデータベースから決定された化合物のライブラリー、またはde novo合成された化合物のライブラリーから選択することができる。
さらなる局面では、本発明は、タウ機能不全を伴う神経障害の予防および治療のための方法であって、それを必要とする対象にタウとFKBP52タンパク質との間の相互作用をモデュレーションする化合物の治療有効量を投与することを含む方法を提供する。該化合物は、本発明のスクリーニング法によって確認されることがある。
本発明のさらなる局面は、診断アッセイ、予後アッセイ、およびモニタリングアッセイに関係する。
材料および方法
抗体および試薬:抗タウmAB(クローンDC25)および抗タウmAB(Tau5)は、それぞれSigmaおよびCalbiochem製であった。抗FKBP52 pAB761は、記載の通りであった(9)。GTPはSigma製であり、ドキシサイクリンはClontech製であった。
H7C2細胞の生成:pTRE2−FKBP52を得るために、ウサギFKBP52をコードするcDNAをpTRE2ベクター(Clontech)のHindIIIおよびAccI制限部位に挿入した。リバーステトラサイクリン制御性トランス活性化因子を発現する市販のPC12 Tet−on細胞系(Clontech)においてリポフェクタミン(商標)(Invitrogen)を使用して100μgのpTR2−FKBP52および10μgのpTK−ヒグロマイシンのトランスフェクションを実施した。安定にトランスフェクションされた細胞を、100μg/mlヒグロマイシンを用いて選択し、個別にスクリーニングした。
タウFKBP52の結合:いくつかの脳領域由来のサイトゾルタンパク質のウエスタンブロットによって示されるように、FKBP52は脳内に広く分布する(図1)。微小管安定性に及ぼすFKBP52の作用にMAPが関与しうるかどうかを検討するために(9)、セファロースビーズに結合したGST−FKBP52を、成体ラット脳から調製された微小管サイトゾルと共にインキュベーションしてGSTプルダウンアッセイを実施した。特異的に結合したタンパク質を、MAP1b、MAP2およびタウに対する抗体を使用した免疫ブロッティングによって分析した。これらの実験条件において、MAP1bまたはMAP2についての免疫反応性は観察されず、タウの免疫反応性が存在した(図2A)。ラット脳ホモジェネートにおいて、タウは、様々なリン酸化度を有する、異なるスプライスアイソフォームを表す複数のバンドとして現れる。いくつかのタウ種は、GST−FKBP52を使用したラット脳サイトゾル微小管のプルダウン実験でも見出された。タウの免疫反応性は、精製GSTを使用した対照では検出されなかった(図2A)。この結合の特異性を確認するために、成体ラット脳サイトゾルの微小管を、FKBP52に対するポリクローナル抗体と共に免疫沈降させた。モノクローナルタウ抗体を用いたウエスタンブロッティングによって免疫沈降物を分析した。タウは、免疫前血清ではなくFKBP52と共に免疫共沈した(図2B)。したがって、タウとFKBP52とは、ラット脳内で複合体を形成する。これらの実験は、FKBP52へのタウの結合が直接的であるかどうかにも、それが追加的な因子を必要とするかどうかにも取組むものではない。これを検討するために、リコンビナントタウ(hT40、E. coliにおいて発現および精製された最長のアイソフォーム)をニトロセルロース上にスポットし、精製リコンビナントFKBP52と共にインキュベーションした。次に、タウによって隔離されたタンパク質を、FKBP52に対するポリクローナル抗体を用いて検出した。図2Cに示すように、FKBP52は、GSTによってではなくタウによって用量依存的に保持された。これらの結果から、FKBP52とタウとの間の直接相互作用が示される。
本発明者らは、この新たに発見されたFKBP52の「抗タウ」活性のおかげで、本来ホルモンステロイドレセプターのモデュレーターとして確認およびクローニングされた(8,22)このタンパク質の機能を再調査することになる。FKPB52は、ペプチジルプロリルイソメラーゼ(「ロタマーゼ」)セグメントを包含する多モジュールタンパク質であって、その機能は、FK506(23)、ラパマイシンおよびいくつかの関係する非免疫抑制誘導体によって遮断される。FKBP52とPin1と(両タンパク質は、ペプチジル−プロリルイソメラーゼ(PPIase)活性および特異的タンパク質−タンパク質相互作用ドメイン(7)を有する)の間には、顕著な構造類似性がある。Pin1 PPIase活性は、アルツハイマー病モデルにおいてリン酸化タウタンパク質の機能を回復させるので(7)、タウとFKBP52との間に観察された相互作用は、アルツハイマー病を含めたタウオパチーの病原にとって意味をもつ可能性がある。FKBP12とは異なり(24)、FKBP52はカルシニューリンと結合しないことから(25)、FKBP52がFK506の免疫抑制能を仲介しないことも忘れてはならない。したがって、非免疫抑制性FK506/ラパマイシン誘導体によるFKBP52のロタマーゼ活性の薬理学的モデュレーションは、ミスフォールディングしたタウの病原作用を防止/軽減するための新規なアプローチを提供する可能性がある。
タウタンパク質凝集物は、複数の神経変性疾患の特徴である。(Delacourte A and Bue L, 2000)。in vitroタウ凝集に関する条件の研究は、酸化条件、ヘパリンなどのポリアニオンおよびアラキドン酸などの脂肪酸による誘導を含めた、異なる実験系につながっている(Barghorn S and Mandelkow E, 2002)。
National BrainBank(GIE NeuroCEB)の利用により得られたヒト前脳前皮質を、10mMトリス、0.32Mサッカロース、1mM DTTおよびプロテアーゼ阻害剤のカクテルを含有する5容量の緩衝液中で均一化した。そのホモジェネートを、14000RPMで4℃で5分間遠心分離した。上清を可溶性画分として使用した。同条件で均一化することによってペレットを可溶化し、不溶性画分として使用した。可溶性画分および不溶性画分中のFKBP52レベルをウエスタンブロッティングによって分析した。可溶性画分および不溶性画分中のFKBP52レベルは、総FKBP52のそれぞれ80%および20%に相当する。
本出願にわたり、様々な参考文献が、本発明が属する技術の現状を説明している。これらの参考文献の開示は、本明細書によって参照により本開示に組入れられる。
Claims (7)
- タウ機能不全を伴う神経障害を有するかまたは有する素因があると考えられる対象を試験する方法であって、
a)上記対象から得られた関心が持たれる試料を:
i)FKBP52タンパク質とタウタンパク質との間の複合体のレベルを測定すること、及び/又は
ii)FKBP52タンパク質の濃度を測定すること、
について分析する工程、
b)FKBP52タンパク質とタウタンパク質との間の複合体のレベルを測定すること及び/又はFKBP52タンパク質の濃度を予め決定されたしきい値と比較することであって、上記比較は、上記対象がタウ機能不全を伴う神経障害を有するか、または有する素因があると考えられるかどうかを示す工程を含み、
ここで、上記関心が持たれる試料が、血液、血清、尿、髄液、または対象の脳組織から得られた生検試料からなる群から選択される、方法。 - タウ機能不全を伴う神経障害が、アルツハイマー病である、請求項1記載の方法。
- 関心が持たれる試料が、髄液からなる群より選択される、請求項1または2記載の方法。
- 関心が持たれる試料を
− その関心が持たれる試料中に存在するFKBP52タンパク質とタウタンパク質との間の複合体と選択的に相互作用可能な結合パートナー、及び/又は
− その関心が持たれる試料中に存在するFKBP52タンパク質と選択的に相互作用可能な結合パートナー及びタウタンパク質と選択的に相互作用可能な別の結合パートナー
と接触させることを含む、請求項1〜3のいずれか記載の方法。 - 結合パートナーが、抗体又はアプタマーである、請求項1〜4のいずれか記載の方法。
- FKBP52タンパク質とタウタンパク質との間の複合体のレベルを、予め決定されたしきい値と比較する工程を含み、ここで、該比較は、対象がタウ機能不全を伴う神経障害を有するか、または有する素因があると考えられるかどうかを示す、請求項1〜5のいずれか記載の方法。
- 対象からの関心が持たれる試料中のFKBP52タンパク質の濃度のレベルを、予め決定されたしきい値と比較することの工程を含み、ここで、該比較は、対象がタウ機能不全を伴う神経障害を有するか、または有する素因があると考えられるかどうかを示す、請求項1〜6のいずれか記載の方法。
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP09305893.1 | 2009-09-24 | ||
EP09305893 | 2009-09-24 | ||
EP10305074.6 | 2010-01-22 | ||
EP10305074 | 2010-01-22 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012530269A Division JP5681719B2 (ja) | 2009-09-24 | 2010-09-24 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016216354A Division JP2017062246A (ja) | 2009-09-24 | 2016-11-04 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2015092186A JP2015092186A (ja) | 2015-05-14 |
JP6039705B2 true JP6039705B2 (ja) | 2016-12-07 |
Family
ID=43032932
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012530269A Active JP5681719B2 (ja) | 2009-09-24 | 2010-09-24 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
JP2015003310A Active JP6039705B2 (ja) | 2009-09-24 | 2015-01-09 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
JP2016216354A Pending JP2017062246A (ja) | 2009-09-24 | 2016-11-04 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
JP2018239614A Pending JP2019069980A (ja) | 2009-09-24 | 2018-12-21 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012530269A Active JP5681719B2 (ja) | 2009-09-24 | 2010-09-24 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016216354A Pending JP2017062246A (ja) | 2009-09-24 | 2016-11-04 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
JP2018239614A Pending JP2019069980A (ja) | 2009-09-24 | 2018-12-21 | タウ機能不全を伴う神経障害を処置するための新規な治療ターゲットとしてのfkbp52−タウ相互作用 |
Country Status (6)
Country | Link |
---|---|
US (6) | US8648044B2 (ja) |
EP (2) | EP2480891B1 (ja) |
JP (4) | JP5681719B2 (ja) |
CN (1) | CN102549438B (ja) |
BR (1) | BR112012008381A2 (ja) |
WO (1) | WO2011045166A1 (ja) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2877397A1 (en) | 2012-07-03 | 2014-01-09 | Washington University | Antibodies to tau |
TW202136296A (zh) | 2014-06-27 | 2021-10-01 | 美商C2N醫療診斷有限責任公司 | 人類化抗-tau抗體 |
JP7429404B2 (ja) * | 2019-08-06 | 2024-02-08 | 慶應義塾 | タウ関連疾患モデルの製造方法 |
Family Cites Families (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
SE8502573D0 (sv) | 1985-05-23 | 1985-05-23 | Jouko Kanakre | Fluorescent lanthanide chelates useful as labels of physiologically active materials |
US5162508A (en) | 1987-12-18 | 1992-11-10 | Compagnie Oris Industrie | Rare earth cryptates, processes for their preparation, synthesis intermediates and application as fluorescent tracers |
US5283173A (en) | 1990-01-24 | 1994-02-01 | The Research Foundation Of State University Of New York | System to detect protein-protein interactions |
DE4011684A1 (de) | 1990-04-06 | 1991-10-10 | Schering Ag | Dtpa-monoamide, diese verbindungen enthaltende pharmazeutische mittel, ihre verwendung und verfahren zu deren herstellung |
US5545806A (en) | 1990-08-29 | 1996-08-13 | Genpharm International, Inc. | Ransgenic non-human animals for producing heterologous antibodies |
CA2121599C (en) * | 1991-10-25 | 2007-04-10 | Marc Mercken | Monoclonal antibodies directed against the microtubule-associated protein tau |
EP0618968B1 (en) * | 1991-12-06 | 1999-10-13 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Tools for the diagnosis and treatment of alzheimer's disease |
US5457186A (en) | 1993-10-13 | 1995-10-10 | Wallac Oy | Luminescent lanthanide chelates with decreased non-radiative energy loss |
US5525490A (en) | 1994-03-29 | 1996-06-11 | Onyx Pharmaceuticals, Inc. | Reverse two-hybrid method |
US5622821A (en) | 1994-06-29 | 1997-04-22 | The Regents Of The University Of California | Luminescent lanthanide chelates and methods of use |
GB9506197D0 (en) * | 1995-03-27 | 1995-05-17 | Hoffmann La Roche | Inhibition of tau-tau association. |
US5637463A (en) | 1995-05-04 | 1997-06-10 | Hoffmann-La Roche Inc. | Method to detect protein-protein interactions |
US5854033A (en) | 1995-11-21 | 1998-12-29 | Yale University | Rolling circle replication reporter systems |
US6740756B1 (en) | 1998-07-07 | 2004-05-25 | Smithkline Beecham Corporation | Fluorescent lanthanide chelates |
JP2002519404A (ja) | 1998-07-07 | 2002-07-02 | スミスクライン・ビーチャム・コーポレイション | 新規蛍光ランタニドキレート |
AU777837B2 (en) * | 2000-01-24 | 2004-11-04 | Innogenetics N.V. | Diagnosis of tauopathies determining tau/phospho-tau ratio |
JP2002040023A (ja) * | 2000-07-28 | 2002-02-06 | Mitsubishi Chemicals Corp | アルツハイマー病の検出方法 |
IL155952A0 (en) * | 2000-11-28 | 2003-12-23 | Wyeth Corp | Expression analysis of fkbp nucleic acids and polypeptides useful in the diagnosis and treatment of prostate cancer |
CA2456175A1 (en) * | 2001-08-06 | 2003-02-20 | Kosan Biosciences, Inc. | Benzoquinone ansamycins |
WO2005040808A1 (en) * | 2003-10-15 | 2005-05-06 | Roche Diagnostics Gmbh | Use of protein fkbp52 as a marker for breast cancer |
AU2005219389A1 (en) | 2004-03-02 | 2005-09-15 | Wyeth | Non-immunosuppressive immunophilin ligands as neuroprotective and/or neuroregenerative agents |
GB0410101D0 (en) * | 2004-05-06 | 2004-06-09 | Leuven K U Res & Dev | Parkinson's disease |
CA2582270A1 (en) * | 2004-11-15 | 2006-05-26 | Blanchette Rockefeller Neurosciences Institute | Abnormalities of phosphatase 2a (pp2a) for diagnosis and treatment of alzheimer's disease |
EP2077853A1 (en) * | 2007-01-29 | 2009-07-15 | Wyeth | Immunophilin ligands and methods for modulating immunophilin and calcium channel activity |
US20080249058A1 (en) * | 2007-04-05 | 2008-10-09 | Erik Roberson | Agents that reduce neuronal overexcitation |
-
2010
- 2010-09-24 EP EP10757413.9A patent/EP2480891B1/en active Active
- 2010-09-24 WO PCT/EP2010/064115 patent/WO2011045166A1/en active Application Filing
- 2010-09-24 BR BR112012008381A patent/BR112012008381A2/pt active Search and Examination
- 2010-09-24 US US13/497,907 patent/US8648044B2/en active Active
- 2010-09-24 CN CN201080044098.0A patent/CN102549438B/zh active Active
- 2010-09-24 JP JP2012530269A patent/JP5681719B2/ja active Active
- 2010-09-24 EP EP16193524.2A patent/EP3139174B1/en active Active
-
2013
- 2013-12-18 US US14/132,832 patent/US9128104B2/en active Active
-
2015
- 2015-01-09 JP JP2015003310A patent/JP6039705B2/ja active Active
- 2015-07-31 US US14/814,674 patent/US9518995B2/en active Active
-
2016
- 2016-10-27 US US15/335,483 patent/US20170056500A1/en not_active Abandoned
- 2016-11-04 JP JP2016216354A patent/JP2017062246A/ja active Pending
-
2018
- 2018-01-15 US US15/871,187 patent/US20180161432A1/en not_active Abandoned
- 2018-12-21 JP JP2018239614A patent/JP2019069980A/ja active Pending
-
2020
- 2020-01-07 US US16/736,316 patent/US20200138951A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
US20140163021A1 (en) | 2014-06-12 |
EP3139174A1 (en) | 2017-03-08 |
US20160047823A1 (en) | 2016-02-18 |
US20200138951A1 (en) | 2020-05-07 |
JP5681719B2 (ja) | 2015-03-11 |
JP2017062246A (ja) | 2017-03-30 |
EP2480891B1 (en) | 2016-11-23 |
US9128104B2 (en) | 2015-09-08 |
US8648044B2 (en) | 2014-02-11 |
US20180161432A1 (en) | 2018-06-14 |
US9518995B2 (en) | 2016-12-13 |
JP2015092186A (ja) | 2015-05-14 |
JP2013506120A (ja) | 2013-02-21 |
US20120302595A1 (en) | 2012-11-29 |
US20170056500A1 (en) | 2017-03-02 |
CN102549438A (zh) | 2012-07-04 |
WO2011045166A9 (en) | 2011-06-16 |
WO2011045166A1 (en) | 2011-04-21 |
BR112012008381A2 (pt) | 2017-06-13 |
EP3139174B1 (en) | 2018-11-07 |
CN102549438B (zh) | 2014-10-29 |
EP2480891A1 (en) | 2012-08-01 |
JP2019069980A (ja) | 2019-05-09 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US9417239B2 (en) | Assay to determine LRRK2 activity in parkinson's disease | |
US11529343B2 (en) | Compositions and methods for identifying and treating dystonia disorders | |
US20200138951A1 (en) | Fkbp52-tau interaction as a novel therapeutical target for treating the neurological disorders involving tau dysfunction | |
US20130273545A1 (en) | Abnormalities of Phosphatase 2A (PP2A) for Diagnosis and Treatment of Alzheimer's Disease | |
KR102120794B1 (ko) | 파킨슨병 진단용 바이오마커, 및 이를 이용한 파킨슨병 진단 방법 | |
US8652785B2 (en) | Method of screening a modulator of endothelial NO synthase comprising the use of heme binding protein 1 | |
US20220187321A1 (en) | Methods for diagnosing alzheimer's disease and other neurological disorders | |
WO2013181064A1 (en) | Method for determining disease severity in tauopathy-related neurodegenerative disorders | |
KR20070084247A (ko) | 알츠하이머 질환의 진단 및 치료를 위한 포스파타제 2a(pp2a)의 비정상 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20150205 |
|
RD04 | Notification of resignation of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7424 Effective date: 20150304 |
|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20151014 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20151020 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20160108 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20160127 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20160621 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20160623 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20161004 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20161104 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6039705 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 Free format text: JAPANESE INTERMEDIATE CODE: R313117 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |