WO1998016557A1 - Dosages pour recepteurs lies a la proteine g - Google Patents
Dosages pour recepteurs lies a la proteine g Download PDFInfo
- Publication number
- WO1998016557A1 WO1998016557A1 PCT/US1996/020510 US9620510W WO9816557A1 WO 1998016557 A1 WO1998016557 A1 WO 1998016557A1 US 9620510 W US9620510 W US 9620510W WO 9816557 A1 WO9816557 A1 WO 9816557A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- receptor
- subunit
- cell
- amino acid
- seq
- Prior art date
Links
- 238000003556 assay Methods 0.000 title description 3
- 238000000034 method Methods 0.000 claims abstract description 60
- 102000034353 G alpha subunit Human genes 0.000 claims abstract description 45
- 108091006099 G alpha subunit Proteins 0.000 claims abstract description 45
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 31
- 229920001184 polypeptide Polymers 0.000 claims abstract description 28
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 28
- 102000030782 GTP binding Human genes 0.000 claims abstract description 17
- 108091000058 GTP-Binding Proteins 0.000 claims abstract description 17
- 108091006027 G proteins Proteins 0.000 claims abstract description 15
- 210000004027 cell Anatomy 0.000 claims description 129
- 102000005962 receptors Human genes 0.000 claims description 108
- 108020003175 receptors Proteins 0.000 claims description 108
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 claims description 54
- 102000005157 Somatostatin Human genes 0.000 claims description 53
- 108010056088 Somatostatin Proteins 0.000 claims description 53
- 229960000553 somatostatin Drugs 0.000 claims description 51
- 102000030621 adenylate cyclase Human genes 0.000 claims description 48
- 108060000200 adenylate cyclase Proteins 0.000 claims description 48
- 230000000694 effects Effects 0.000 claims description 43
- 108020004707 nucleic acids Proteins 0.000 claims description 39
- 102000039446 nucleic acids Human genes 0.000 claims description 39
- 150000007523 nucleic acids Chemical class 0.000 claims description 39
- 102100023803 Somatostatin receptor type 3 Human genes 0.000 claims description 34
- 150000001413 amino acids Chemical class 0.000 claims description 29
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 22
- 102100023806 Somatostatin receptor type 5 Human genes 0.000 claims description 20
- 102000014303 Amyloid beta-Protein Precursor Human genes 0.000 claims description 19
- 108010079054 Amyloid beta-Protein Precursor Proteins 0.000 claims description 19
- 150000001875 compounds Chemical class 0.000 claims description 17
- 210000004899 c-terminal region Anatomy 0.000 claims description 16
- 239000003446 ligand Substances 0.000 claims description 16
- 210000004881 tumor cell Anatomy 0.000 claims description 14
- 230000003993 interaction Effects 0.000 claims description 13
- 125000001433 C-terminal amino-acid group Chemical group 0.000 claims description 12
- 125000000539 amino acid group Chemical group 0.000 claims description 11
- 108010058828 Parathyroid Hormone Receptors Proteins 0.000 claims description 10
- 102000006461 Parathyroid Hormone Receptors Human genes 0.000 claims description 10
- 230000002401 inhibitory effect Effects 0.000 claims description 10
- 230000012010 growth Effects 0.000 claims description 8
- 102000014415 Muscarinic acetylcholine receptor Human genes 0.000 claims description 5
- 108050003473 Muscarinic acetylcholine receptor Proteins 0.000 claims description 5
- 108090000445 Parathyroid hormone Proteins 0.000 claims description 5
- 239000013603 viral vector Substances 0.000 claims description 5
- 108010031792 IGF Type 2 Receptor Proteins 0.000 claims description 4
- 102000038460 IGF Type 2 Receptor Human genes 0.000 claims description 4
- 241000124008 Mammalia Species 0.000 claims description 4
- 102000009346 Adenosine receptors Human genes 0.000 claims description 3
- 108050000203 Adenosine receptors Proteins 0.000 claims description 3
- 102000008873 Angiotensin II receptor Human genes 0.000 claims description 3
- 108050000824 Angiotensin II receptor Proteins 0.000 claims description 3
- 108010001789 Calcitonin Receptors Proteins 0.000 claims description 3
- 102100038520 Calcitonin receptor Human genes 0.000 claims description 3
- 102000010180 Endothelin receptor Human genes 0.000 claims description 3
- 108050001739 Endothelin receptor Proteins 0.000 claims description 3
- 108700023400 Platelet-activating factor receptors Proteins 0.000 claims description 3
- 108091008874 T cell receptors Proteins 0.000 claims description 3
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 claims description 3
- 102000016715 Transforming Growth Factor beta Receptors Human genes 0.000 claims description 3
- 108010092867 Transforming Growth Factor beta Receptors Proteins 0.000 claims description 3
- 229960003638 dopamine Drugs 0.000 claims description 3
- 210000003061 neural cell Anatomy 0.000 claims description 3
- 102000030769 platelet activating factor receptor Human genes 0.000 claims description 3
- 108060003345 Adrenergic Receptor Proteins 0.000 claims description 2
- 102000017910 Adrenergic receptor Human genes 0.000 claims description 2
- 208000024827 Alzheimer disease Diseases 0.000 claims description 2
- 206010041067 Small cell lung cancer Diseases 0.000 claims description 2
- 102100029329 Somatostatin receptor type 1 Human genes 0.000 claims description 2
- 208000000587 small cell lung carcinoma Diseases 0.000 claims description 2
- 108010082379 somatostatin receptor type 1 Proteins 0.000 claims description 2
- 230000001131 transforming effect Effects 0.000 claims description 2
- 108090000680 somatostatin receptor 5 Proteins 0.000 claims 2
- 101710192647 Somatostatin receptor type 3 Proteins 0.000 claims 1
- 108090000166 Thrombin receptors Proteins 0.000 claims 1
- 102000003790 Thrombin receptors Human genes 0.000 claims 1
- RZWIIPASKMUIAC-VQTJNVASSA-N thromboxane Chemical compound CCCCCCCC[C@H]1OCCC[C@@H]1CCCCCCC RZWIIPASKMUIAC-VQTJNVASSA-N 0.000 claims 1
- 238000002560 therapeutic procedure Methods 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract description 3
- 229940124606 potential therapeutic agent Drugs 0.000 abstract 1
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 description 36
- IVOMOUWHDPKRLL-UHFFFAOYSA-N UNPD107823 Natural products O1C2COP(O)(=O)OC2C(O)C1N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-UHFFFAOYSA-N 0.000 description 36
- 229940095074 cyclic amp Drugs 0.000 description 36
- 101000829138 Homo sapiens Somatostatin receptor type 3 Proteins 0.000 description 33
- 230000015572 biosynthetic process Effects 0.000 description 25
- 108020004414 DNA Proteins 0.000 description 20
- 108050001286 Somatostatin Receptor Proteins 0.000 description 19
- 102000011096 Somatostatin receptor Human genes 0.000 description 19
- 101000829153 Homo sapiens Somatostatin receptor type 5 Proteins 0.000 description 18
- 230000000638 stimulation Effects 0.000 description 18
- 238000010168 coupling process Methods 0.000 description 17
- 238000005859 coupling reaction Methods 0.000 description 17
- 238000001890 transfection Methods 0.000 description 14
- 239000013598 vector Substances 0.000 description 14
- 239000002299 complementary DNA Substances 0.000 description 13
- 108090000623 proteins and genes Proteins 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 12
- 239000013612 plasmid Substances 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- INAPMGSXUVUWAF-GCVPSNMTSA-N [(2r,3s,5r,6r)-2,3,4,5,6-pentahydroxycyclohexyl] dihydrogen phosphate Chemical compound OC1[C@H](O)[C@@H](O)C(OP(O)(O)=O)[C@H](O)[C@@H]1O INAPMGSXUVUWAF-GCVPSNMTSA-N 0.000 description 11
- 238000011282 treatment Methods 0.000 description 11
- 239000012636 effector Substances 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 10
- 206010028980 Neoplasm Diseases 0.000 description 9
- 101000829127 Homo sapiens Somatostatin receptor type 2 Proteins 0.000 description 8
- 102100023802 Somatostatin receptor type 2 Human genes 0.000 description 8
- GGYTXJNZMFRSLX-DFTNLTQTSA-N somatostatin-28 Chemical compound N([C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H]1C(N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CSSC1)C(O)=O)[C@@H](C)O)[C@@H](C)O)=O)C(=O)[C@@H]1CCCN1C(=O)[C@H](CC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CO GGYTXJNZMFRSLX-DFTNLTQTSA-N 0.000 description 8
- 230000008878 coupling Effects 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 150000003905 phosphatidylinositols Chemical class 0.000 description 7
- 108010081690 Pertussis Toxin Proteins 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 6
- 230000001225 therapeutic effect Effects 0.000 description 6
- 230000007306 turnover Effects 0.000 description 6
- 230000001404 mediated effect Effects 0.000 description 5
- 230000004614 tumor growth Effects 0.000 description 5
- 108010049048 Cholera Toxin Proteins 0.000 description 4
- 102000009016 Cholera Toxin Human genes 0.000 description 4
- 102000003982 Parathyroid hormone Human genes 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 208000025688 early-onset autosomal dominant Alzheimer disease Diseases 0.000 description 4
- 208000015756 familial Alzheimer disease Diseases 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000000199 parathyroid hormone Substances 0.000 description 4
- 229960001319 parathyroid hormone Drugs 0.000 description 4
- 230000004936 stimulating effect Effects 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 3
- 102000000844 Cell Surface Receptors Human genes 0.000 description 3
- 108010001857 Cell Surface Receptors Proteins 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 208000009889 Herpes Simplex Diseases 0.000 description 3
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 3
- 102100023801 Somatostatin receptor type 4 Human genes 0.000 description 3
- 238000000692 Student's t-test Methods 0.000 description 3
- 102000014384 Type C Phospholipases Human genes 0.000 description 3
- 108010079194 Type C Phospholipases Proteins 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 230000003190 augmentative effect Effects 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000010369 molecular cloning Methods 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000003362 replicative effect Effects 0.000 description 3
- 108010064556 somatostatin receptor subtype-4 Proteins 0.000 description 3
- DEQANNDTNATYII-OULOTJBUSA-N (4r,7s,10s,13r,16s,19r)-10-(4-aminobutyl)-19-[[(2r)-2-amino-3-phenylpropanoyl]amino]-16-benzyl-n-[(2r,3r)-1,3-dihydroxybutan-2-yl]-7-[(1r)-1-hydroxyethyl]-13-(1h-indol-3-ylmethyl)-6,9,12,15,18-pentaoxo-1,2-dithia-5,8,11,14,17-pentazacycloicosane-4-carboxa Chemical compound C([C@@H](N)C(=O)N[C@H]1CSSC[C@H](NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](CC=2C3=CC=CC=C3NC=2)NC(=O)[C@H](CC=2C=CC=CC=2)NC1=O)C(=O)N[C@H](CO)[C@H](O)C)C1=CC=CC=C1 DEQANNDTNATYII-OULOTJBUSA-N 0.000 description 2
- PUDHBTGHUJUUFI-SCTWWAJVSA-N (4r,7s,10s,13r,16s,19r)-10-(4-aminobutyl)-n-[(2s,3r)-1-amino-3-hydroxy-1-oxobutan-2-yl]-19-[[(2r)-2-amino-3-naphthalen-2-ylpropanoyl]amino]-16-[(4-hydroxyphenyl)methyl]-13-(1h-indol-3-ylmethyl)-6,9,12,15,18-pentaoxo-7-propan-2-yl-1,2-dithia-5,8,11,14,17-p Chemical compound C([C@H]1C(=O)N[C@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(N[C@@H](CSSC[C@@H](C(=O)N1)NC(=O)[C@H](N)CC=1C=C2C=CC=CC2=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(N)=O)=O)C(C)C)C1=CC=C(O)C=C1 PUDHBTGHUJUUFI-SCTWWAJVSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 2
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 description 2
- 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 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- 108010016076 Octreotide Proteins 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 208000013403 hyperactivity Diseases 0.000 description 2
- 229960000367 inositol Drugs 0.000 description 2
- -1 inositol phosphates Chemical class 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 108010021336 lanreotide Proteins 0.000 description 2
- 238000001638 lipofection Methods 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 210000002850 nasal mucosa Anatomy 0.000 description 2
- 229960002700 octreotide Drugs 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- 230000001177 retroviral effect Effects 0.000 description 2
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- APIXJSLKIYYUKG-UHFFFAOYSA-N 3 Isobutyl 1 methylxanthine Chemical compound O=C1N(C)C(=O)N(CC(C)C)C2=C1N=CN2 APIXJSLKIYYUKG-UHFFFAOYSA-N 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- YEELWQSXYBJVSV-UWJYBYFXSA-N Ala-Cys-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YEELWQSXYBJVSV-UWJYBYFXSA-N 0.000 description 1
- KQESEZXHYOUIIM-CQDKDKBSSA-N Ala-Lys-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KQESEZXHYOUIIM-CQDKDKBSSA-N 0.000 description 1
- AWNAEZICPNGAJK-FXQIFTODSA-N Ala-Met-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(O)=O AWNAEZICPNGAJK-FXQIFTODSA-N 0.000 description 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- QOIGKCBMXUCDQU-KDXUFGMBSA-N Ala-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C)N)O QOIGKCBMXUCDQU-KDXUFGMBSA-N 0.000 description 1
- 101710137189 Amyloid-beta A4 protein Proteins 0.000 description 1
- 101710151993 Amyloid-beta precursor protein Proteins 0.000 description 1
- 102100022704 Amyloid-beta precursor protein Human genes 0.000 description 1
- HJVGMOYJDDXLMI-AVGNSLFASA-N Arg-Arg-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCCNC(N)=N HJVGMOYJDDXLMI-AVGNSLFASA-N 0.000 description 1
- RVDVDRUZWZIBJQ-CIUDSAMLSA-N Arg-Asn-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O RVDVDRUZWZIBJQ-CIUDSAMLSA-N 0.000 description 1
- SIFXMYAHXJGAFC-WDSKDSINSA-N Arg-Asp Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(O)=O SIFXMYAHXJGAFC-WDSKDSINSA-N 0.000 description 1
- OZNSCVPYWZRQPY-CIUDSAMLSA-N Arg-Asp-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O OZNSCVPYWZRQPY-CIUDSAMLSA-N 0.000 description 1
- HJAICMSAKODKRF-GUBZILKMSA-N Arg-Cys-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O HJAICMSAKODKRF-GUBZILKMSA-N 0.000 description 1
- HPKSHFSEXICTLI-CIUDSAMLSA-N Arg-Glu-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HPKSHFSEXICTLI-CIUDSAMLSA-N 0.000 description 1
- XLWSGICNBZGYTA-CIUDSAMLSA-N Arg-Glu-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O XLWSGICNBZGYTA-CIUDSAMLSA-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
- YHZQOSXDTFRZKU-WDSOQIARSA-N Arg-Trp-Leu Chemical compound C1=CC=C2C(C[C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N)=CNC2=C1 YHZQOSXDTFRZKU-WDSOQIARSA-N 0.000 description 1
- QLSRIZIDQXDQHK-RCWTZXSCSA-N Arg-Val-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QLSRIZIDQXDQHK-RCWTZXSCSA-N 0.000 description 1
- DAPLJWATMAXPPZ-CIUDSAMLSA-N Asn-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O DAPLJWATMAXPPZ-CIUDSAMLSA-N 0.000 description 1
- JZDZLBJVYWIIQU-AVGNSLFASA-N Asn-Glu-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JZDZLBJVYWIIQU-AVGNSLFASA-N 0.000 description 1
- GLWFAWNYGWBMOC-SRVKXCTJSA-N Asn-Leu-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O GLWFAWNYGWBMOC-SRVKXCTJSA-N 0.000 description 1
- ZYPWIUFLYMQZBS-SRVKXCTJSA-N Asn-Lys-Lys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZYPWIUFLYMQZBS-SRVKXCTJSA-N 0.000 description 1
- AMRANMVXQWXNAH-ZLUOBGJFSA-N Asp-Cys-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CS)NC(=O)[C@@H](N)CC(O)=O AMRANMVXQWXNAH-ZLUOBGJFSA-N 0.000 description 1
- FMWHSNJMHUNLAG-FXQIFTODSA-N Asp-Cys-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CCCN=C(N)N FMWHSNJMHUNLAG-FXQIFTODSA-N 0.000 description 1
- QQXOYLWJQUPXJU-WHFBIAKZSA-N Asp-Cys-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O QQXOYLWJQUPXJU-WHFBIAKZSA-N 0.000 description 1
- UJGRZQYSNYTCAX-SRVKXCTJSA-N Asp-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UJGRZQYSNYTCAX-SRVKXCTJSA-N 0.000 description 1
- PWAIZUBWHRHYKS-MELADBBJSA-N Asp-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CC(=O)O)N)C(=O)O PWAIZUBWHRHYKS-MELADBBJSA-N 0.000 description 1
- NALWOULWGHTVDA-UWVGGRQHSA-N Asp-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NALWOULWGHTVDA-UWVGGRQHSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 101100315624 Caenorhabditis elegans tyr-1 gene Proteins 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 108091028075 Circular RNA Proteins 0.000 description 1
- 108010069514 Cyclic Peptides Proteins 0.000 description 1
- 102000001189 Cyclic Peptides Human genes 0.000 description 1
- WTEACWBAULENKE-SRVKXCTJSA-N Cys-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CS)N WTEACWBAULENKE-SRVKXCTJSA-N 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical class CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 102000001301 EGF receptor Human genes 0.000 description 1
- 108060006698 EGF receptor Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- BDAGIHXWWSANSR-UHFFFAOYSA-N Formic acid Chemical group OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 1
- 102000040442 G-alpha family Human genes 0.000 description 1
- 108091072318 G-alpha family Proteins 0.000 description 1
- ITYRYNUZHPNCIK-GUBZILKMSA-N Glu-Ala-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O ITYRYNUZHPNCIK-GUBZILKMSA-N 0.000 description 1
- WOSRKEJQESVHGA-CIUDSAMLSA-N Glu-Arg-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O WOSRKEJQESVHGA-CIUDSAMLSA-N 0.000 description 1
- CKOFNWCLWRYUHK-XHNCKOQMSA-N Glu-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)O)N)C(=O)O CKOFNWCLWRYUHK-XHNCKOQMSA-N 0.000 description 1
- CYHBMLHCQXXCCT-AVGNSLFASA-N Glu-Asp-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CYHBMLHCQXXCCT-AVGNSLFASA-N 0.000 description 1
- ZZIFPJZQHRJERU-WDSKDSINSA-N Glu-Cys-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O ZZIFPJZQHRJERU-WDSKDSINSA-N 0.000 description 1
- NKLRYVLERDYDBI-FXQIFTODSA-N Glu-Glu-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O NKLRYVLERDYDBI-FXQIFTODSA-N 0.000 description 1
- OAGVHWYIBZMWLA-YFKPBYRVSA-N Glu-Gly-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)NCC(O)=O OAGVHWYIBZMWLA-YFKPBYRVSA-N 0.000 description 1
- HILMIYALTUQTRC-XVKPBYJWSA-N Glu-Gly-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HILMIYALTUQTRC-XVKPBYJWSA-N 0.000 description 1
- VXQOONWNIWFOCS-HGNGGELXSA-N Glu-His-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CCC(=O)O)N VXQOONWNIWFOCS-HGNGGELXSA-N 0.000 description 1
- BBBXWRGITSUJPB-YUMQZZPRSA-N Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](N)CCC(O)=O BBBXWRGITSUJPB-YUMQZZPRSA-N 0.000 description 1
- OQXDUSZKISQQSS-GUBZILKMSA-N Glu-Lys-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O OQXDUSZKISQQSS-GUBZILKMSA-N 0.000 description 1
- UERORLSAFUHDGU-AVGNSLFASA-N Glu-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N UERORLSAFUHDGU-AVGNSLFASA-N 0.000 description 1
- MIIGESVJEBDJMP-FHWLQOOXSA-N Glu-Phe-Tyr Chemical compound C([C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 MIIGESVJEBDJMP-FHWLQOOXSA-N 0.000 description 1
- CQAHWYDHKUWYIX-YUMQZZPRSA-N Glu-Pro-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O CQAHWYDHKUWYIX-YUMQZZPRSA-N 0.000 description 1
- JSIQVRIXMINMTA-ZDLURKLDSA-N Glu-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H](N)CCC(O)=O JSIQVRIXMINMTA-ZDLURKLDSA-N 0.000 description 1
- DTLLNDVORUEOTM-WDCWCFNPSA-N Glu-Thr-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(O)=O DTLLNDVORUEOTM-WDCWCFNPSA-N 0.000 description 1
- UCZXXMREFIETQW-AVGNSLFASA-N Glu-Tyr-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O UCZXXMREFIETQW-AVGNSLFASA-N 0.000 description 1
- JPXNYFOHTHSREU-UWVGGRQHSA-N Gly-Arg-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CN JPXNYFOHTHSREU-UWVGGRQHSA-N 0.000 description 1
- 108010050006 Gly-Asp-Gly-Arg Proteins 0.000 description 1
- CEXINUGNTZFNRY-BYPYZUCNSA-N Gly-Cys-Gly Chemical compound [NH3+]CC(=O)N[C@@H](CS)C(=O)NCC([O-])=O CEXINUGNTZFNRY-BYPYZUCNSA-N 0.000 description 1
- QSVCIFZPGLOZGH-WDSKDSINSA-N Gly-Glu-Ser Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QSVCIFZPGLOZGH-WDSKDSINSA-N 0.000 description 1
- YIFUFYZELCMPJP-YUMQZZPRSA-N Gly-Leu-Cys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(O)=O YIFUFYZELCMPJP-YUMQZZPRSA-N 0.000 description 1
- NTBOEZICHOSJEE-YUMQZZPRSA-N Gly-Lys-Ser Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O NTBOEZICHOSJEE-YUMQZZPRSA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- RVKIPWVMZANZLI-UHFFFAOYSA-N H-Lys-Trp-OH Natural products C1=CC=C2C(CC(NC(=O)C(N)CCCCN)C(O)=O)=CNC2=C1 RVKIPWVMZANZLI-UHFFFAOYSA-N 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- LQGCNWWLGGMTJO-ULQDDVLXSA-N His-Met-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N LQGCNWWLGGMTJO-ULQDDVLXSA-N 0.000 description 1
- 208000037147 Hypercalcaemia Diseases 0.000 description 1
- 102000003746 Insulin Receptor Human genes 0.000 description 1
- 108010001127 Insulin Receptor Proteins 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- QOOWRKBDDXQRHC-BQBZGAKWSA-N L-lysyl-L-alanine Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN QOOWRKBDDXQRHC-BQBZGAKWSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- FMEICTQWUKNAGC-YUMQZZPRSA-N Leu-Gly-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O FMEICTQWUKNAGC-YUMQZZPRSA-N 0.000 description 1
- XWOBNBRUDDUEEY-UWVGGRQHSA-N Leu-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 XWOBNBRUDDUEEY-UWVGGRQHSA-N 0.000 description 1
- DDVHDMSBLRAKNV-IHRRRGAJSA-N Leu-Met-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O DDVHDMSBLRAKNV-IHRRRGAJSA-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
- KCXUCYYZNZFGLL-SRVKXCTJSA-N Lys-Ala-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O KCXUCYYZNZFGLL-SRVKXCTJSA-N 0.000 description 1
- GJJQCBVRWDGLMQ-GUBZILKMSA-N Lys-Glu-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O GJJQCBVRWDGLMQ-GUBZILKMSA-N 0.000 description 1
- ZUGVARDEGWMMLK-SRVKXCTJSA-N Lys-Ser-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN ZUGVARDEGWMMLK-SRVKXCTJSA-N 0.000 description 1
- UASDAHIAHBRZQV-YUMQZZPRSA-N Met-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(O)=O)CCCNC(N)=N UASDAHIAHBRZQV-YUMQZZPRSA-N 0.000 description 1
- OHMKUHXCDSCOMT-QXEWZRGKSA-N Met-Asn-Val Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O OHMKUHXCDSCOMT-QXEWZRGKSA-N 0.000 description 1
- STLBOMUOQNIALW-BQBZGAKWSA-N Met-Gly-Cys Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H](CS)C(O)=O STLBOMUOQNIALW-BQBZGAKWSA-N 0.000 description 1
- PBOUVYGPDSARIS-IUCAKERBSA-N Met-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(O)=O)CC(C)C PBOUVYGPDSARIS-IUCAKERBSA-N 0.000 description 1
- 108010046068 N-Acetyllactosamine Synthase Proteins 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- 101100342977 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) leu-1 gene Proteins 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- JVTMTFMMMHAPCR-UBHSHLNASA-N Phe-Ala-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 JVTMTFMMMHAPCR-UBHSHLNASA-N 0.000 description 1
- HHOOEUSPFGPZFP-QWRGUYRKSA-N Phe-Asn-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O HHOOEUSPFGPZFP-QWRGUYRKSA-N 0.000 description 1
- SCKXGHWQPPURGT-KKUMJFAQSA-N Phe-Lys-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O SCKXGHWQPPURGT-KKUMJFAQSA-N 0.000 description 1
- QARPMYDMYVLFMW-KKUMJFAQSA-N Phe-Pro-Glu Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=CC=C1 QARPMYDMYVLFMW-KKUMJFAQSA-N 0.000 description 1
- APZNYJFGVAGFCF-JYJNAYRXSA-N Phe-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)Cc1ccccc1)C(C)C)C(O)=O APZNYJFGVAGFCF-JYJNAYRXSA-N 0.000 description 1
- HXOLCSYHGRNXJJ-IHRRRGAJSA-N Pro-Asp-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HXOLCSYHGRNXJJ-IHRRRGAJSA-N 0.000 description 1
- MGDFPGCFVJFITQ-CIUDSAMLSA-N Pro-Glu-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MGDFPGCFVJFITQ-CIUDSAMLSA-N 0.000 description 1
- XQHGISDMVBTGAL-ULQDDVLXSA-N Pro-His-Phe Chemical compound C([C@@H](C(=O)[O-])NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H]1[NH2+]CCC1)C1=CC=CC=C1 XQHGISDMVBTGAL-ULQDDVLXSA-N 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 101001135767 Rattus norvegicus Parathyroid hormone Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 208000007014 Retinitis pigmentosa Diseases 0.000 description 1
- VBKBDLMWICBSCY-IMJSIDKUSA-N Ser-Asp Chemical compound OC[C@H](N)C(=O)N[C@H](C(O)=O)CC(O)=O VBKBDLMWICBSCY-IMJSIDKUSA-N 0.000 description 1
- BNFVPSRLHHPQKS-WHFBIAKZSA-N Ser-Asp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O BNFVPSRLHHPQKS-WHFBIAKZSA-N 0.000 description 1
- LRZLZIUXQBIWTB-KATARQTJSA-N Ser-Lys-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LRZLZIUXQBIWTB-KATARQTJSA-N 0.000 description 1
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 1
- PQEQXWRVHQAAKS-SRVKXCTJSA-N Ser-Tyr-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CO)N)CC1=CC=C(O)C=C1 PQEQXWRVHQAAKS-SRVKXCTJSA-N 0.000 description 1
- ILVGMCVCQBJPSH-WDSKDSINSA-N Ser-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CO ILVGMCVCQBJPSH-WDSKDSINSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- IRKWVRSEQFTGGV-VEVYYDQMSA-N Thr-Asn-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O IRKWVRSEQFTGGV-VEVYYDQMSA-N 0.000 description 1
- QWMPARMKIDVBLV-VZFHVOOUSA-N Thr-Cys-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O QWMPARMKIDVBLV-VZFHVOOUSA-N 0.000 description 1
- CQNFRKAKGDSJFR-NUMRIWBASA-N Thr-Glu-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O CQNFRKAKGDSJFR-NUMRIWBASA-N 0.000 description 1
- DXPURPNJDFCKKO-RHYQMDGZSA-N Thr-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DXPURPNJDFCKKO-RHYQMDGZSA-N 0.000 description 1
- YEGMNOHLZNGOCG-UBHSHLNASA-N Trp-Asn-Asn Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O YEGMNOHLZNGOCG-UBHSHLNASA-N 0.000 description 1
- DVIIYMVCSUQOJG-QEJZJMRPSA-N Trp-Glu-Asp Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O DVIIYMVCSUQOJG-QEJZJMRPSA-N 0.000 description 1
- BODHJXJNRVRKFA-BZSNNMDCSA-N Tyr-Cys-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BODHJXJNRVRKFA-BZSNNMDCSA-N 0.000 description 1
- KLQPIEVIKOQRAW-IZPVPAKOSA-N Tyr-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O KLQPIEVIKOQRAW-IZPVPAKOSA-N 0.000 description 1
- UDNYEPLJTRDMEJ-RCOVLWMOSA-N Val-Asn-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N UDNYEPLJTRDMEJ-RCOVLWMOSA-N 0.000 description 1
- QGFPYRPIUXBYGR-YDHLFZDLSA-N Val-Asn-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N QGFPYRPIUXBYGR-YDHLFZDLSA-N 0.000 description 1
- BMGOFDMKDVVGJG-NHCYSSNCSA-N Val-Asp-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BMGOFDMKDVVGJG-NHCYSSNCSA-N 0.000 description 1
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 1
- JKHXYJKMNSSFFL-IUCAKERBSA-N Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(O)=O)CCCCN JKHXYJKMNSSFFL-IUCAKERBSA-N 0.000 description 1
- NZGOVKLVQNOEKP-YDHLFZDLSA-N Val-Phe-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N NZGOVKLVQNOEKP-YDHLFZDLSA-N 0.000 description 1
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 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 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 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 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000003416 augmentation Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000002051 biphasic effect Effects 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000035605 chemotaxis Effects 0.000 description 1
- 238000012411 cloning technique Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000023077 detection of light stimulus Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000007783 downstream signaling Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- YMAWOPBAYDPSLA-UHFFFAOYSA-N glycylglycine Chemical compound [NH3+]CC(=O)NCC([O-])=O YMAWOPBAYDPSLA-UHFFFAOYSA-N 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000000148 hypercalcaemia Effects 0.000 description 1
- 208000030915 hypercalcemia disease Diseases 0.000 description 1
- 210000003016 hypothalamus Anatomy 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 229960002437 lanreotide Drugs 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 208000006155 precocious puberty Diseases 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 108010070643 prolylglutamic acid Proteins 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 230000035943 smell Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- OGBMKVWORPGQRR-UMXFMPSGSA-N teriparatide Chemical compound C([C@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)[C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 OGBMKVWORPGQRR-UMXFMPSGSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 230000002463 transducing effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000003146 transient transfection Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4722—G-proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
- C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/566—Immunoassay; Biospecific binding assay; Materials therefor using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- 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
- G01N2333/4719—G-proteins
-
- 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
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
- G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
- G01N2333/726—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
-
- 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
Definitions
- the field of the invention is GTP-binding proteins and the receptors to which they link.
- G proteins are the best characterized and the most versatile. They elicit biological functions which include hormone signalling, neurotransmission, chemotaxis, and perception of light, smell, and taste. G proteins couple to various cell surface receptors (G-linked receptors) and activate various intracellular effectors. Each G protein is made up of a G ⁇ subunit and a G ⁇ subunit. The specificity of G proteins' coupling to receptors and downstream signalling molecules is conferred by the various G ⁇ subunits.
- the G ⁇ molecules are classified into two categories: one is a class of sensory-organ-specific G proteins (e.g., G ⁇ t , G ⁇ olf , and G ⁇ gust ) , and the other is a less tissue-specific class consisting of G ⁇ g , the G family (i.e., G ⁇ i;L , G ⁇ i2 , G ⁇ i3 , Ga ol , G ⁇ o2 , and G ⁇ 2 ) , the G ⁇ 12 family (i.e., G ⁇ 12 and G ⁇ 13 ) , and the G ⁇ q family (i.e, G ⁇ cron, G ⁇ ⁇ , G ⁇ 14 , and G ⁇ 16 ) . It is likely that more members of each class will be discovered.
- G ⁇ g the G family
- the G family i.e., G ⁇ i;L , G ⁇ i2 , G ⁇ i3 , Ga ol , G ⁇ o2 , and G ⁇ 2
- G ⁇ s -based chimeric system for identifying the G ⁇ subunit of a G protein to which a given G-linked receptor couples.
- a series of G ⁇ s / ⁇ ⁇ chimeras (G ⁇ ⁇ : any G ⁇ subunit except G ⁇ g ) can be made with a first amino acid sequence corresponding to a region of G ⁇ s (SEQ ID NO: 21) encompassing G g 's residues 236-356, followed by a second amino acid sequence 4-30 amino acids long and corresponding to a segment of G ⁇ x , which segment ends at (and includes) G ⁇ ⁇ 's C-terminal residue.
- the first amino acid sequence should contain the effector portion of G ⁇ s , and preferably will contain residues 1-389 of SEQ ID NO: 21.
- the second amino acid sequence should contain the receptor-coupling portion of G ⁇ ⁇ , and preferably is 4 or 5 amino acids in length (e.g., as represented by SEQ ID NOs:22-30).
- two identical samples of cells are provided, wherein the cells co-express a given G-linked receptor and a given G ⁇ s / ⁇ ⁇ chimera.
- the second sample of cells is contacted with a ligand of the G- linked receptor.
- AC activity as anisfested by the rate of cAMP formation, is measured in both samples of cells.
- a significant increase in cAMP formation in the second sample as compared to the first sample indicates that that particular G ⁇ ⁇ can couple to the receptor.
- Cells co-expressing a given G-linked receptor and a given G ⁇ g / ⁇ ⁇ chimera can be established by introducing into the cells a recombinant nucleic acid construct permitting expression of the receptor and a second recombinant nucleic acid construct permitting expression of the chimera.
- recombinant is meant that the nucleic acid (or polypeptide) molecule is the product of artificial genetic manipulation.
- a G ⁇ s / ⁇ ⁇ chimera is a polypeptide which includes the AC-coupling portion (e.g., amino acid residues 236-356) of G ⁇ s (SEQ ID NO: 21) as well as the receptor-coupling portion of G ⁇ x .
- the receptor-coupling portion can be 4-30 amino acids long and usually corresponds to the extreme C-terminal region of G ⁇ ⁇ .
- the chimeric polypeptide can also include an additional peptide sequence such as one that serves as an epitope tag, so long as the additional sequence does not interfere with the functioning of the chimera.
- G- linked receptor is meant any naturally occurring cell surface receptor, or any functional recombinant variant thereof, that couples to a G protein.
- ligand is meant any molecule that binds and activates a receptor.
- a ligand can be, for example, the natural, physiological activator of the receptor (e.g., a hormone), a biologically active analogue thereof, or an antibody which binds to and thereby activates the receptor.
- the chimeras of the invention can also be used in a method of screening compounds for their ability to modulate the interaction between a given G-linked receptor and the G ⁇ (i.e., G ⁇ ⁇ ) subunit of a non-G s G protein known to couple to the receptor.
- G ⁇ i.e., G ⁇ ⁇
- two identical samples of cells are provided, wherein the cells co-express the G-linked receptor and a G ⁇ g / ⁇ ⁇ chimera. Both samples of cells are contacted with a ligand of the G-linked receptor. The second sample is additionally contacted with a test compound. cAMP formation is then measured in both samples.
- a significant decrease (or increase) of the cAMP level in the second sample as compared to the first sample indicates that the compound is capable of inhibiting (or enhancing) the interaction between the G-linked receptor and that particular G ⁇ ⁇ .
- signal-transducing output is meant the end result of the signalling initiated by a liganded G-linked receptor. Such an end result can be, for example, cell growth inhibition, cell proliferation, or secretion of a protein.
- G ⁇ chimeric polypeptide containing the sequence of a G ⁇ linking to a desirable effector, the receptor-coupling region (e.g., the 4-30 residues at the C-terminal end) of which sequence is replaced with that of a G ⁇ to which the G-linked receptor normally couples.
- the receptor-coupling region e.g., the 4-30 residues at the C-terminal end
- Such a chimeric polypeptide can be employed in a method of therapy for a condition associated with the function or lack of function of that receptor in a patient's cells.
- the G ⁇ s / ⁇ Q polypeptide can be introduced into the target cell by introducing into the cell a recombinant nucleic acid construct that permits expression of the chimeric polypeptide.
- a construct can, for instance, be derived from a herpes simplex viral vector, or any other vector able to transfect neural cells.
- SST somatostatin
- SSTR SST receptors
- Somatostatin is a 14 amino acid cyclic peptide hormone which was originally isolated from the hypothalamus.
- Biologically active analogues of SST include, but are not limited to, (1) naturally occurring analogues, such as SST-28 (FEBS Lett. 282: 363-367, 1991) and SST-25 (Gen CompEndocinol 81: 365-372, 1991); and (2) artificial compounds, such as octreotide (New Engl. J. Med. 334: 246-254, 1995), RC-160 (Buscail et al., PNAS 92: 1580-1584, 1995), RC-160-I and RC-160-II (Cancer Res. 54: 5895-5901, 1994), SMS 201-995 (Kubota et al., J.
- Another method of inhibiting tumor growth is useful for tumor cells the growth of which is stimulated via an endogenous, hyperactive G-linked receptor.
- endogenous is meant that the receptor is expressed in the cell absent any artificial genetic manipulation.
- hyperactive is meant that the G-linked receptor is more active, or active for a longer period of time, than it is in a normal cell. Hyperactivity of a G-linked receptor can be caused by, for example, certain mutations in the receptor's peptide sequence, an unusually high level of the receptor's ligand, and/or a ligand that dissociates from the receptor at a rate lower than normal.
- One can then introduce into the tumor cell a G ⁇ 12 or G 13 chimeric molecule, the C-terminal 5 residues of which are replaced with those of the G ⁇ that the hyperactive receptor normally couples to.
- the hyperactivity of the receptor is transduced via the G ⁇ 12 or G ⁇ 13 chimeric molecule to downstream growth-inhibitory effectors, which counteract at least in part growth-stimulatory signals normally transduced by the receptor and its cognate G protein.
- the chimera will transduce a signal that results in apoptosis of the tumor cell.
- the chimeric molecule can be introduced into the target cell in vivo , in vitro , or ex vivo in a carrier such as saline and/or liposomes. It can also be expressed by a recombinant nucleic acid construct that has been introduced into the cell.
- Fig. 1 is a schematic representation of the G ⁇ s chimeras constructed in the study.
- G ⁇ s wt denotes wild-type G ⁇ s .
- Sequences of the last 5 C-terminal residues of the chimeras are illustrated, and referred to as SEQ ID NOs : 22-31. These sequences are identical between G ⁇ i:L and G ⁇ i2 , between G ⁇ ol and G ⁇ o2 , and between G ⁇ q and G ⁇ ⁇ .
- Fig. 2A is a bar graph showing the effects of SST on cholera toxin (CTX) -stimulated AC activity in cells expressing SSTR3.
- CCTX cholera toxin
- Cells were transfected with 0.125 ⁇ g of pCMV6-SSTR3 and 0.125 ⁇ g of pCMV6 vector.
- cells were treated for 30 min with or without 1 ⁇ M SST, in the presence of (1) 1 mM IBMX, or (2) lmM IBMX plus 250 ng/ml CTX.
- cAMP formation was subsequently measured. All values are "means ⁇ S.E.” of quadruplicated experiments.
- Fig. 2B & Fig. 2C are bar graphs showing the effects of SST on cAMP formation in cells expressing a G ⁇ s chimera with (Fig. 2C) or without (Fig. 2B) SSTR3.
- Cells were transfected with 0.125 ⁇ g of plasmid encoding a G ⁇ g chimera and 0.125 ⁇ g of either pCMV6-SSTR3 (Fig. 2C) or pCMV6 (Fig. 2B) .
- Fig. 2C pCMV6-SSTR3
- Fig. 2B pCMV6
- AC activity levels are represented as percentage relative to the basal AC activity level in cells expressing G ⁇ g / ⁇ i;L alone. All values are "means ⁇ S.E.” of quadruplicated experiments. Similar results were found at least three times for each chimera.
- Fig. 2D is a bar graph converted from Fig. 2B, showing the ratios of cAMP levels in the presence vs. absence of SST in transfected cells.
- Fig. 3A & Fig. 3B are bar graphs showing the effects of SST on AC activity in cells expressing a Ga s chimera with (Fig. 3A) or without (Fig. 3B) SSTR3.
- Cells were transfected with 0.125 ⁇ g of plasmid encoding a G ⁇ chimera and either 0.125 ⁇ g of pCMV6-SSTR3 (Fig. 3A) or pCMV6 (Fig. 3B) .
- AC activity levels are represented as percentage relative to the basal AC activity level in cells expressing G ⁇ g / ⁇ q alone. All values are "means ⁇ S.E.” of quadruplicated experiments. Similar results were found at least three times for each chimera.
- Fig. 3C is a bar graph showing the effects of SST on cAMP formation in cells expressing SSTR3 and a G ⁇ s chimera derived from the G ⁇ i or G family. All the indicated chimeras were tested in parallel. Experiments were performed as described in the legend for Figs. 2A and 2B. All values are "means ⁇ S.E.” of quadruplicated experiments. Similar results were found at least three times for each chimera.
- Fig. 3D is a bar graph converted from Fig. 3C, showing the ratios of cAMP levels in the presence vs. absence of SST in transfected cells.
- Fig. 4A is a bar graph showing the stimulation of inositol phosphate (IP) production in cells transfected with (1) 0.125 ⁇ g of pCMV6-SSTR3, and (2) 0.125 ⁇ g of plasmid encoding the intact G ⁇ 16 .
- IP inositol phosphate
- cells were treated for 5 min with or without 1 ⁇ M SST, and IP production was measured.
- PTX pertussis toxin
- cells were treated with 10 ng/ml PTX for 3 h and with SST as described above.
- Fig. 4B is a bar graph showing the stimulation of IP production in cells transfected with (1) 0.125 ⁇ g of pCMV6-SSTR3, and (2) 0.125 ⁇ g of plasmid encoding intact G ⁇ 14 . Experiments were performed as described in Fig. 4A's legend.
- Fig. 4C is a bar graph showing the stimulation of IP production in cells transfected with (1) 0.125 ⁇ g of pCMV6-SSTR3, and (2) 0.125 ⁇ g of plasmid encoding intact G ⁇ . Experiments were performed as described in Fig. 4A's legend.
- Fig. 4D is a bar graph showing the stimulation of IP production in cells transfected with (1) 0.125 ⁇ g of plasmid encoding parathyroid hormone receptor (PTHR) , and (2) 0.125 ⁇ g of plasmid encoding intact G ⁇ . Experiments were performed as described in Fig. 4A's legend.
- PTHR parathyroid hormone receptor
- Fig. 5A is a bar graph showing the effects of SST on AC activity in cells expressing a SSTR and G ⁇ g / ⁇ 12 .
- Cells were transfected with (1) 0.125 ⁇ g of plasmid encoding G ⁇ s / ⁇ 12 , and (2) 0.125 ⁇ g of pCMV6-SSTRl, pCMV6- SSTR2, pCMV6-SSTR3, pCMV6-SSTR5, pCDNAI-SSTR4 , or pCMV6.
- cells were stimulated with 1 ⁇ M SST and cAMP formation was measured.
- Fig. 5B is a bar graph converted from Fig. 5A, showing the ratios of cAMP levels in the presence vs. absence of SST.
- Fig. 5C is a bar graph showing the effects of SST on AC activity in cells expressing a SSTR and G ⁇ g / ⁇ 13 .
- Cells were transfected with (1) 0.125 ⁇ g of plasmid encoding G ⁇ g / ⁇ 13 , and (2) 0.125 ⁇ g of pCMV6-SSTRl, pCMV6- SSTR2, pCMV6-SSTR3, pCMV6-SSTR5, pCDNAI-SSTR4 , or pCMV6.
- cells were stimulated with 1 ⁇ M SST and cAMP formation was measured.
- Fig. 5D is a bar graph converted from Fig. 5C, showing the ratios of cAMP levels in the presence vs. absence of SST.
- G ⁇ subtype coupling can be assigned for any given G-linked receptor.
- the following examples are meant to illustrate, but not limit, the methods of the present invention.
- Other suitable modifications and adaptations of the conditions which are obvious to those skilled in the art are within the scope and spirit of the invention.
- genetically engineered variants of G-linked receptors can be substituted for the naturally occurring receptors.
- Standard transfection techniques other than the lipofection technique illustrated below, e.g., calcium phosphate precipitation, biolistic transfer, DEAE- Dextran, and viral-vector methods, can also be employed in the invention.
- COS cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum and antibiotics, as described previously (Ikezu et al., J. Biol. Chem. 270: 29224-29228, 1995) Transient transfection was performed by lipofection as previously described (Ikezu et al. , J. Biol. Chem. 270: 29224-29228, 1995) .
- 2xl0 4 cells were seeded onto a 24-well plate and cultured in complete growth medium for 24 h.
- the cells were subsequently transfected with 0.25 ⁇ g of plasmid and 1 ⁇ l LipofectAMINETM (GIBCO-BRL) for another 24 h in serum-free DMEM, and cultured in complete growth medium for an additional 24 h. Measurement of AC activity
- Intact-cell AC activity was assessed by measuring cAMP formation as described previously (Ikezu et al., J. Biol. Chem. 270: 29224-29228, 1995).
- cells were labeled with 6 ⁇ Ci/ l of [ 3 H]adenine (Du Pont-NEN) for 24 h, and then treated with ligands of the G-linked receptor of interest (e.g. , somatostatin-14 for a somatostatin receptor) and 1 mM IBMX (3-isobutyl-l-methylxanthine) for 30 min.
- the resultant radioactive cAMP was separated on two-step ion-exchange columns. Specific accumulation of cAMP was expressed as [cAMP/(ADP + ATP)] x 10 3 , which represents intact-cell AC activity.
- Statistical analysis was performed with Student's t test. Measurement of PI Turnover
- PI phosphatidyl inositol turnover was assessed by measuring IP (inositol phosphates) production.
- IP inositol phosphates
- 4xl0 4 cells were seeded onto a 24-well plate, cultured in complete growth medium for 24 h, and transfected for 24 h as described above. The culture medium was replaced with the labeling medium [inositol-free RPMI supplemented with dialyzed fetal calf serum and 10 ⁇ Ci/ml of [ 3 H]myo-inositol (Du Pont-NEN)].
- the cells were washed four times with inositol-free RPMI and treated with 1 ⁇ l somatostatin (SST) in inositol-free RPMI at 37 °C for 5 min. After discarding the medium, the cells in 0.2 ml fresh medium were lysed on the plate by 0.8 ml of ice-cold 12.5% (final concentration: 10%) TCA, and the lysate was put on ice for 20 min before centrifugation. Supernatant of the lysate was mixed well with 1 ml of saturated diethyl ether to extract acid.
- SST somatostatin
- constructs (designated pCMV6-SSTRl, pCMV6-SSTR2, pCMV6-SSTR3, and pCMV6-SSTR5) , all of which were derived from a pCMV6 vector (the SSTRl and 2 constructs: pCMV6b; the SSTR3 and 5 constructs: pCMV6c) , contain the SSTR coding sequences under the transcriptional control of the cytomegalovirus promoter.
- the SSTR4 expression construct (pcDNAI-SSTR4) was made by inserting the SSTR4 cDNA (Bito et al. , J. Biol. Chem. 269: 12722-12730, 1994) in pBluescript (Strategene) into pcDNAI (Invitrogen) .
- the G ⁇ s chimeras were constructed as follows. First, PCR was performed to add Aflll and Xbal sites at the 3' end of the wild type G ⁇ g cDNA using the following two primers: ATCTGGAATAACAGATGGCTGC (SEQ ID N0:1) and
- the PCR product was digested with Bglll and Xbal, and subcloned into pcDNAI-G ⁇ s (i.e., the original plasmid containing the wild type G ⁇ s cDNA) which had been predigested with the same enzymes .
- the resultant construct designated G ⁇ g -AX, was sequenced to confirm the presence of Aflll and Xbal sites. Subsequently, the construct was digested with Aflll and Xbal, and ligated with two synthetic oligonucleotides to add sequence encoding the carboxyl-terminal five residues of a non-G ⁇ g subunit.
- the oligonucleotides were:
- CTAGATTAACACAAACCGATGTATC SEQ ID NO: 10 (for G ⁇ s / ⁇ z ) ;
- G ⁇ g Stimulation of G ⁇ g , but not any other G ⁇ , results in an increase in adenylyl cyclase (AC) activity.
- AC adenylyl cyclase
- G ⁇ g / ⁇ x G ⁇ ⁇ : any type of G ⁇ except G ⁇ g ) chimeras wherein the last five C-terminal residues of the G ⁇ s polypeptide are replaced with those of G ⁇ x . If a receptor couples to G ⁇ x , it will, upon binding to its ligand, recognize and activate the G ⁇ g / ⁇ ⁇ chimera, thereby resulting in G ⁇ s -mediated AC stimulation in the cell.
- G ⁇ g / ⁇ x chimeras consisting of G ⁇ g 1-389 (which lacks the original five C-terminal residues of G ⁇ g ) and the five C-terminal residues of each known G ⁇ were constructed.
- the five C-terminal residues are identical between G ⁇ i:L and G ⁇ i2 , between G ⁇ Ql and G ⁇ o2 , and between G ⁇ q and G ⁇ ⁇ ;L .
- G ⁇ s / ⁇ i;L G ⁇ s / ⁇ i3 , G ⁇ g / ⁇ 0 , G ⁇ g / ⁇ z , G ⁇ B / ⁇ q , G ⁇ s / ⁇ 12 , G ⁇ g / ⁇ 13 , G ⁇ g / ⁇ 14 , and G ⁇ s / ⁇ 16 , respectively (Fig. 1).
- the residues 1-389 (SEQ ID N0:21) of G ⁇ g are the following:
- the experimental strategy was to transiently express a G ⁇ g / ⁇ ⁇ cDNA along with a SSTR cDNA, and then to compare AC activities in the presence and absence of SST. If treatment with SST promotes cAMP formation only in cells expressing the SSTR and a given G ⁇ g / ⁇ ⁇ , one can assume the linkage of the SSTR to that G ⁇ g / ⁇ ⁇ and therefore to that G ⁇ ⁇ .
- the G ⁇ g chimeras were each expressed as a 52-kDa protein at similar levels in COS cells, consistent with expected molecular weight.
- G ⁇ s / ⁇ ⁇ chimeras where G ⁇ ⁇ is derived from the G ⁇ i family (G ⁇ G ⁇ 0 , and G ⁇ z ) were tested for their ability to transduce AC-stimulatory signal initiated by SST-bound SSTR3.
- SSTR3 has been shown to function as a G ⁇ coupled receptor and to suppress AC activity in various cell types (Yasuda et al., J. Biol. Chem. 267: 20422-20428, 1992; Ya ada et al., Mol. Endocrinol. 6: 2136-2142, 1992; Kaupmann et al., FEBS Lett.
- G ⁇ proteins known to inhibit AC are members of the G ⁇ j ⁇ family, which include the Ga ' s (i.e., G ⁇ ilf G ⁇ i2 , G ⁇ i3 ) , the G ⁇ 0 's (i.e., G ⁇ ol and G ⁇ o2 ) , and Ga z (Wong et al., Nature 351: 63-65, 1991), the present data suggest that SSTR3 may inhibit cAMP formation exclusively through the G ⁇ i 's.
- Fig. 2A Since SST significantly reduced cAMP formation when SSTR3 was transfected without G ⁇ g / ⁇ 14 or G ⁇ g / ⁇ 16 (Fig. 2A) , it is conceivable that the net stimulation of AC by SSTR3 through these two chimeras may have been considerably larger than what was observed. In contrast, in cells co- expressing SSTR3 and G ⁇ ./ ⁇ , no stimulation of AC was observed under the same conditions.
- Figs. 3C and 3D show results of the experiments wherein the linkage of SSTR3 to chimeras derived from the G ⁇ and G ⁇ families were examined in parallel. Again, the results demonstrated that SSTR3 may link to Ga 14 and G ⁇ 16 , in addition to the G ⁇ i's, but not to any other members of the G ⁇ i and G ⁇ families .
- inability of a G-linked receptor to couple to a given G ⁇ s / ⁇ ⁇ indicates the inability of the receptor to recognize the C terminus of that particular G ⁇ x , and therefore rules out the coupling between the receptor and the intact G ⁇ ⁇ .
- the present study suggests for the first time that SSTR3 may not couple to G ⁇ Q , G ⁇ z , G ⁇ q , G ⁇ 11; G ⁇ 12 , or Ga 13 (see below for G ⁇ 12 and G ⁇ 13 ) .
- ability of a G- linked receptor to couple to a given G ⁇ s / ⁇ ⁇ indicates that the receptor is capable of coupling to that particular G ⁇ ⁇ .
- the present chimeric system is extremely useful in identifying potential receptor-G ⁇ linkage, especially for G ⁇ 's which have less established signal-transducing effectors and which therefore are less amenable to assaying.
- the present system can be employed to identify G ⁇ 12 - or G ⁇ 13 -coupled receptors.
- G ⁇ 12 and G ⁇ 13 have been implicated in pivotal cellular functions (Voyno-Yasenetskaya et al., Oncogene 9: 2559-2565, 1994 and Voyno-Yasenetskaya et al., J. Biol. Chem. 269: 4721-4724, 1994), receptors to which they couple remain elusive.
- G ⁇ 12 and G ⁇ 13 couple to any of the 5 known subtypes of SSTR's
- SST-induced cAMP formation was measured in cells co-expressing a given SSTR and either G ⁇ g /G ⁇ 12 or G ⁇ s / GQ: 13 - F gs.
- 5A-5D shows that G ⁇ s /G ⁇ 12 was activated by SSTR2 , 4, and 5 in the order of SSTR5 >> SSTR2 ⁇ SSTR4 , while G ⁇ g /G ⁇ 13 was activated almost exclusively by SSTR5.
- the stimulation of G ⁇ g /G ⁇ 12 and G ⁇ s /G ⁇ 13 by liganded SSTR5 yielded a more than 5 fold increase in the cAMP level (Figs. 5B and 5D) .
- Table I shows that, in the presence of G ⁇ s / ⁇ 12 , the stimulation of cAMP formation by SSTR5 and SSTR2 is SST-dosage-dependent and biphasic. At low SST concentrations, cAMP formation was slightly but reproducibly inhibited, whereas at higher concentrations, cAMP formation was strongly stimulated. It is conceivable that the former effect may be mediated by
- SST-28 had a more potent effect on the function of SSTR5 than the naturally occurring SST (i.e., SST-14) , regarding both of their inhibitory and stimulatory effects.
- the present system can also be used to investigate proteins with a G-linked-receptor-like structure (e.g., with multiple transmembrane domains) but having unknown functions.
- the system can also be used to investigate proteins which have only a single-transmembrane domain but which are suspected of being a G-linked receptor.
- insulin-like growth factor II receptor Murayama et al., J. Biol. Chem. 265: 17456- 17462, 1990
- amyloid precursor protein APP
- sperm ⁇ - 1, 4-galactosyltransferase Gong et al., Science, 269: 1718-1721, 1995.
- epidermal growth factor receptor Sun et al., Proc. Natl. Acad. Sci.
- One aspect of the present invention is a method of identifying a compound that can modulate the interaction between a G-linked receptor and the G ⁇ subunit of a non- G s G protein known to couple to the receptor.
- two samples of cells are provided, both of which express (a) the receptor of interest, and (b) a chimeric polypeptide containing amino acid residues 1-389 of G ⁇ g (SEQ ID NO: 21) followed by the C-terminal 5 amino acid residues of the non-G ⁇ s G ⁇ subunit known to couple to the receptor.
- a ligand of the G-linked receptor is administered to both cell samples.
- the second cell Prior to, subsequent to, or at the same time as the ligand administration, the second cell is contacted with a candidate compound.
- the activity of adenylyl cyclase in each cell sample is determined and compared as described above.
- a statistically significant (i.e., p ⁇ 0.05 in Student's t test) change of the AC activity in the second cell sample as compared to the first cell sample indicates that the compound may be capable of modulating the interaction between the G-linked receptor and the coupling G ⁇ subunit.
- a statistically significant decrease of the AC activity in the compound-contacted cells will suggest that the compound may block the interaction.
- the efficacy of the compound can be confirmed by a second assay using the full length G ⁇ subunit instead of the chimera.
- Cell lines that can be used in connection with this method include those of mammalian origin, such as COS cells and HEK 293 cells (American Type Culture Collection) . Maintenance and transfection of such cells can be performed using well known methods. Proteins (a) and (b) (see above) can be introduced into the target cells via transfection of nucleic acid constructs encoding them. Techniques for making nucleic acid constructs are well known in the art (see, e.g., Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1989) ; examples of such techniques have been illustrated above.
- G-linked receptors of interest include, but are not limited to, those described in U.S. Patent No.5, 559, 209 , herein incorporated by reference (e.g., insulin-like growth factor II receptor, muscarinic acetylcholine receptor, ⁇ 2 -adrenergic receptor, adenosine receptor, thrombin receptor, transforming growth factor ⁇ receptor, T cell receptor, PTH/PTHrP receptor, calcitonin receptor, endothelin receptor, angiotensin receptor, platelet activating factor receptor, thromboxane A 2 receptor, any of the somatostatin receptors, D 2 -dopamine receptor, ⁇ -butyric acid receptor) , and amyloid protein precursor (APP) .
- insulin-like growth factor II receptor e.g., insulin-like growth factor II receptor, muscarinic acetylcholine receptor, ⁇ 2 -adrenergic receptor, adenosine receptor, thro
- APP has at least 10 isoforms, one of which (APP 695 ) is preferentially expressed in neuronal tissue (Sandbrink et al., J. Biol. Chem. 269: 1510, 1994).
- the construction of a baculovirus construct containing the APP 695 cDNA has been described (Nishimoto et al., Nature 362: 75-79, 1993) .
- Similar cloning techniques can be employed to create APP 695 mammalian expression constructs based on mammalian expression vectors such as pCDNAI and pCMV6.
- Constitutively active variants of the G-linked receptors can also be used in the present screening method, eliminating the need for their ligands.
- three constitutively active APP 695 mutants designated Ile-APP, Phe-APP, and Gly-APP, have been identified in familial Alzheimer's Disease patients (Ya atsuji et al., Science 272: 1349-1352, 1996; and references therein) .
- These three mutants have mis-sense mutations in which Val 642 in the transmembrane domain of APP 695 is replaced by lie, Phe, or Gly, respectively.
- the chimeras of the invention can alternatively be used in a method of altering the signal-transducing output of a G-linked receptor.
- Abnormalities of G-linked receptor functions have been implicated in many significant diseases such as familial Alzheimer's disease (Nishimoto et al. , Nature 362: 75-79, 1993; Yamatsuji et al., Science 272: 1349-1352, 1996; Okamoto et al. , The EMBO J. 15: 3769-3777, 1996; Ikezu et al., The EMBO J.
- Amyloid protein precursor (APP) , a G-linked cell surface receptor, has been shown to be mutated and constitutively active in at least some forms of familial Alzheimer's Disease (Okamoto et al., The EMBO J. 15: 3769-3777, 1996; and references therein). APP is known to couple to G Q , the activation of which inhibits adenylyl cyclase (Okamoto et al., The EMBO J. 15: 3769- 3777, 1996 and references therein) .
- the present invention provides a method for augmenting adenylyl cyclase activity in brain neurons of a mammal, and preferably, of a familial Alzheimer's patient.
- a G ⁇ s subunit in which the C- terminal 5 aa residues are replaced with those of G ⁇ Q is introduced into the brain neurons of the mammal.
- This chimeric G ⁇ molecule will compete with the endogenous G ⁇ 0 for the binding of APP, and upon binding to APP, will transduce stimulatory signals to adenylyl cyclase, thereby counteracting the inhibitory signals transduced by native G 0 .
- This chimeric molecule can be introduced into the target cell by overexpressing within the target cell a nucleic acid construct comprising a promoter sequence operably linked to a sequence encoding the protein.
- the nucleic acid construct is typically derived from a non-replicating linear or circular DNA or RNA vector, or from an autonomously replicating plasmid or viral vector; or the construct is integrated into the host genome.
- These nucleic acid constructs can be made with methods well known in the art (see, e.g., Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, New York, 1989) . Any vector that can transfect a brain neuron may be used in the method of the invention.
- a preferred vector is a herpes simplex viral (HSV) vector or an appropriately modified version of this vector.
- HSV herpes simplex viral
- a therapeutic composition containing this vector may be used alone or in a mixture, or in chemical combination, with one or more materials, including other proteins or recombinant vectors that increase the biological stability of the recombinant vectors, or with materials that increase the therapeutic composition's ability to penetrate the target tissue selectively.
- the therapeutic compositions of the invention is typically administered in a pharmaceutically acceptable carrier (e.g., physiological saline), which is selected on the basis of the mode and route of administration, and standard pharmaceutical practice.
- a pharmaceutically acceptable carrier e.g., physiological saline
- Suitable pharmaceutical carriers, as well as pharmaceutical necessities for use in pharmaceutical formulations, are described in Remington 's Pharmaceutical Sciences , a standard reference text in this field, and in the USP/NF.
- the therapeutic compositions of the invention can be administered in dosages determined to be appropriate by one skilled in the art. It is expected that the dosages will vary, depending upon the pharmacokinetic and pharmacodynamic characteristics of the particular agent, and its mode and route of administration, as well as the age, weight, and health of the recipient; the nature and extent of the disease; the frequency and duration of the treatment; the type of, if any, concurrent therapy; and the desired effect.
- the therapeutic compositions may be administered to a patient by any appropriate mode, e.g., via applying drops or spray onto the nasal mucosa, or via injection into the nasal mucosa, as determined by one skilled in the art. Alternatively, it may be desired to administer the treatment surgically to the target tissue. The treatments of the invention may be repeated as needed, as determined by one skilled in the art.
- the invention includes a method of inhibiting tumor growth by expressing an exogenously introduced SSTR5 protein, e.g., a recombinant protein comprising (a) SSTR5, or (b) a biologically active fragment thereof, in a tumor cell.
- SSTR5 protein e.g., a recombinant protein comprising (a) SSTR5, or (b) a biologically active fragment thereof
- Recombinant G ⁇ 12 or G ⁇ 13 polypeptides can also be introduced into the target cell.
- the recombinant SSTR5 present on the cell surface will be stimulated and will thereby inhibit growth of the tumor cell via endogenous or recombinant G ⁇ 12 and G ⁇ 13 .
- This aspect of the invention is useful in cancer treatments using SST-related drugs (i.e., SST or SST analogues) .
- SST-related drugs i.e., SST or SST analogues
- Such treatments frequently lead to loss of SSTR's naturally expressed on cancer cells, thereby desensitizing the cells to the SST-related drugs.
- Introduction of recombinant SSTR5 into the cancer cells solves this problem, at least temporarily; further transfetions may be necessary to maintain the effect, if the recombinant SSTR5 is lost as well. All cancers, including highly malignant ones such as pancreatic cancer and small cell lung cancer, can be treated by the present method.
- the recombinant SSTR5 protein can be introduced into the cancer cells by overexpressing within the cells a nucleic acid construct comprising a mammalian promoter sequence operably linked to a sequence encoding the protein.
- the construct primarily targets fast-proliferating cells, and can, for example, be derived from retroviral, adenoviral, adeno-associated- viral, or herpes simplex viral vectors, or any appropriately modified versions of these vectors.
- Retroviral vectors are particularly appropriate, as they selectively integrate into the genome of replicating cells, such as tumor cells.
- Gly Gly Gin Arg Asp Gin Arg Arg Lys Trp lie Gin Cys Phe Asn Asp 225 230 235 240
- Val Thr Ala lie lie Phe Val Val Ala Ser Ser Ser Tyr Asn Met Val
- Lys Ser Lys lie Glu Asp Tyr Phe Pro Glu Phe Ala Arg Tyr Thr Thr 305 310 315 320
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Endocrinology (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
L'invention porte sur des polypeptides chimères dérivés des sous-unités Gα de diverses protéines G, et sur des procédés d'utilisation des polypeptides chimères dans des thérapies et dans le criblage d'agents thérapeutiques potentiels.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US2834096P | 1996-10-11 | 1996-10-11 | |
US60/028,340 | 1996-10-11 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1998016557A1 true WO1998016557A1 (fr) | 1998-04-23 |
Family
ID=21842899
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1996/020510 WO1998016557A1 (fr) | 1996-10-11 | 1996-12-16 | Dosages pour recepteurs lies a la proteine g |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO1998016557A1 (fr) |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999014344A1 (fr) * | 1997-09-13 | 1999-03-25 | Glaxo Group Limited | Chimeres de proteines g |
WO2000006722A1 (fr) * | 1998-07-28 | 2000-02-10 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | Clonage par interaction de genes a base de cellules eukaryiotes |
WO2001036481A1 (fr) * | 1999-11-17 | 2001-05-25 | Yung Hou Wong | Nouvelles protéine g-alpha à plus grande promiscuité avec les gprc |
WO2001072802A1 (fr) * | 2000-03-28 | 2001-10-04 | Shanghai Biowindow Gene Development Inc. | Nouveau polypeptide, proteine humaine de liaison 14 d'une proteine precurseur de l'amyloide, et polynucleotide codant pour ce polypeptide |
WO2001079438A3 (fr) * | 2000-03-29 | 2002-02-28 | Biowindow Gene Dev Inc | Nouveau polypeptide, proteine humaine de liaison 9 d'une proteine precurseur de l'amyloide, et polynucleotide codant pour ce polypeptide |
WO2002024867A3 (fr) * | 2000-09-22 | 2003-08-14 | Univ Aarhus | Nouvelles compositions et nouvelles methodes pour le diagnostic et le traitement des lymphomes et des leucemies |
WO2003027276A3 (fr) * | 2001-09-24 | 2004-02-12 | Univ Aarhus | Nouvelles compositions et procedes relatifs aux lymphomes et aux leucemies |
DE10233516A1 (de) * | 2002-07-23 | 2004-02-12 | Aventis Pharma Deutschland Gmbh | Verfahren zur Identifizierung von Substanzen |
WO2005047318A1 (fr) * | 2003-11-11 | 2005-05-26 | Astrazeneca Ab | Variant d'epissage de gnal et ses applications |
EP1328663A4 (fr) * | 2000-09-28 | 2005-06-01 | New England Medical Center Inc | Dosages d'identifications de recepteurs presentant des alterations de signaux de transduction |
US7067277B1 (en) | 1999-12-23 | 2006-06-27 | H. Lundbeck A/S | Chimeric G proteins and uses thereof |
US7674584B2 (en) | 2004-09-30 | 2010-03-09 | Ge Healthcare Uk Limited | Method for measuring binding of a test compound to a G-protein coupled receptor |
WO2017129998A1 (fr) * | 2016-01-29 | 2017-08-03 | Heptares Therapeutics Limited | Protéines g |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5559209A (en) * | 1993-02-18 | 1996-09-24 | The General Hospital Corporation | Regulator regions of G proteins |
US5578451A (en) * | 1993-02-18 | 1996-11-26 | The General Hospital Corporation | Methods and systems for screening potential alzheimer's disease therapeutics |
-
1996
- 1996-12-16 WO PCT/US1996/020510 patent/WO1998016557A1/fr active Application Filing
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5559209A (en) * | 1993-02-18 | 1996-09-24 | The General Hospital Corporation | Regulator regions of G proteins |
US5578451A (en) * | 1993-02-18 | 1996-11-26 | The General Hospital Corporation | Methods and systems for screening potential alzheimer's disease therapeutics |
Non-Patent Citations (5)
Title |
---|
CELL, 06 March 1992, Vol. 68, BERLOT C.H. et al., "Identification of Effector-Activating Residues of Gsa", pages 911-922. * |
J. BIOL. CHEM., 18 February 1994, Vol. 269, No. 7, VOYNO-YASENETSKAYA T. et al., "Galpha13 Stimulates Na-H Exchange", pages 4721-4724. * |
MOLECULAR PHARMACOLOGY, April 1994, Vol. 45, No. 4, LAW S.F. et al., "Gialpha1 Selectively Couples Somatostatin Receptor Subtype 3 to Adenylyl Cyclase: Identification of the Functional Domains of This Alpha Subunit Necessary for Mediating the Inhibition by Somatostatin of cAMP Formation", pages 587-590. * |
NATURE, 20 May 1993, Vol. 363, CONKLIN B.R. et al., "Substitution of Three Amino Acids Switches Receptor Specificity of Gqalpha to That Gialpha", pages 274-276. * |
SCIENCE, 31 May 1996, Vol. 272, YAMATSUJI T. et al., "G Protein-Mediated Neuronal DNA Fragmentation Induced by Familial Alzheimer's Disease-Associated Mutants of APP", pages 1349-1352. * |
Cited By (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1533379A1 (fr) * | 1997-09-13 | 2005-05-25 | Glaxo Group Limited | Chiméras de Protéine G |
US6509447B1 (en) | 1997-09-13 | 2003-01-21 | Glaxo Welcome Inc. | G protein chimeras and methods of screening compounds |
WO1999014344A1 (fr) * | 1997-09-13 | 1999-03-25 | Glaxo Group Limited | Chimeres de proteines g |
WO2000006722A1 (fr) * | 1998-07-28 | 2000-02-10 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | Clonage par interaction de genes a base de cellules eukaryiotes |
WO2001036481A1 (fr) * | 1999-11-17 | 2001-05-25 | Yung Hou Wong | Nouvelles protéine g-alpha à plus grande promiscuité avec les gprc |
US6462178B1 (en) | 1999-11-17 | 2002-10-08 | Yong Hou Wong | G protein |
US7067277B1 (en) | 1999-12-23 | 2006-06-27 | H. Lundbeck A/S | Chimeric G proteins and uses thereof |
WO2001072802A1 (fr) * | 2000-03-28 | 2001-10-04 | Shanghai Biowindow Gene Development Inc. | Nouveau polypeptide, proteine humaine de liaison 14 d'une proteine precurseur de l'amyloide, et polynucleotide codant pour ce polypeptide |
WO2001079438A3 (fr) * | 2000-03-29 | 2002-02-28 | Biowindow Gene Dev Inc | Nouveau polypeptide, proteine humaine de liaison 9 d'une proteine precurseur de l'amyloide, et polynucleotide codant pour ce polypeptide |
WO2002024867A3 (fr) * | 2000-09-22 | 2003-08-14 | Univ Aarhus | Nouvelles compositions et nouvelles methodes pour le diagnostic et le traitement des lymphomes et des leucemies |
EP1328663A4 (fr) * | 2000-09-28 | 2005-06-01 | New England Medical Center Inc | Dosages d'identifications de recepteurs presentant des alterations de signaux de transduction |
WO2003027276A3 (fr) * | 2001-09-24 | 2004-02-12 | Univ Aarhus | Nouvelles compositions et procedes relatifs aux lymphomes et aux leucemies |
DE10233516A1 (de) * | 2002-07-23 | 2004-02-12 | Aventis Pharma Deutschland Gmbh | Verfahren zur Identifizierung von Substanzen |
WO2005047318A1 (fr) * | 2003-11-11 | 2005-05-26 | Astrazeneca Ab | Variant d'epissage de gnal et ses applications |
US7674584B2 (en) | 2004-09-30 | 2010-03-09 | Ge Healthcare Uk Limited | Method for measuring binding of a test compound to a G-protein coupled receptor |
CN108699123A (zh) * | 2016-01-29 | 2018-10-23 | 赫普泰雅治疗有限公司 | G蛋白 |
GB2558968A (en) * | 2016-01-29 | 2018-07-25 | Heptares Therapeutics Ltd | G Proteins |
WO2017129998A1 (fr) * | 2016-01-29 | 2017-08-03 | Heptares Therapeutics Limited | Protéines g |
JP2019507590A (ja) * | 2016-01-29 | 2019-03-22 | ヘプタレス セラピューティックス リミテッド | Gタンパク質 |
US10738287B2 (en) | 2016-01-29 | 2020-08-11 | Heptares Therapeutics Limited | G proteins |
AU2017212788B2 (en) * | 2016-01-29 | 2021-07-29 | Nxera Pharma Uk Limited | G proteins |
GB2558968B (en) * | 2016-01-29 | 2021-09-29 | Heptares Therapeutics Ltd | G Proteins |
US11339383B2 (en) | 2016-01-29 | 2022-05-24 | Heptares Therapeutics Limited | G proteins |
JP2022084574A (ja) * | 2016-01-29 | 2022-06-07 | ヘプタレス セラピューティックス リミテッド | Gタンパク質 |
CN108699123B (zh) * | 2016-01-29 | 2023-12-08 | 赫普泰雅治疗有限公司 | G蛋白 |
JP7524234B2 (ja) | 2016-01-29 | 2024-07-29 | ヘプタレス セラピューティックス リミテッド | Gタンパク質 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Jones et al. | Golf: an olfactory neuron specific-G protein involved in odorant signal transduction | |
JPH06508765A (ja) | グルカゴン様ペプチド−1(glp−1)に対するレセプター | |
US20200247866A1 (en) | Teneurin c-terminal associated peptides (tcap) and methods and uses thereof | |
ZA200705031B (en) | Y4 selective receptor agonists for therapeutic interventions | |
WO1998016557A1 (fr) | Dosages pour recepteurs lies a la proteine g | |
US20100150909A1 (en) | PTHrP-DERIVED MODULATORS OF SMOOTH MUSCLE PROLIFERATION | |
TWI227780B (en) | Methods for identifying compounds for regulating muscle mass or function using corticotropin releasing factor receptors | |
US20050272660A1 (en) | Polypeptide derivatives of parathyroid hormone (PTH) | |
US20020147170A1 (en) | Constitutively active, hypersensitive, and nonfunctional recepors as novel therapeutic agents | |
MXPA05012546A (es) | Materiales y metodos que se relacionan con los oligomeros receptores acoplados a la proteina g. | |
US7022815B1 (en) | Polypeptide derivatives of parathyroid hormone (PTH) | |
JP4723144B2 (ja) | 副甲状腺ホルモン(pth)のポリペプチド誘導体 | |
Ho et al. | The amino terminus of Gαz is required for receptor recognition, whereas its α4/β6 loop is essential for inhibition of adenylyl cyclase | |
CA2426779A1 (fr) | Procede d'identification de composes pour la regulation de la masse ou fonction musculaire au moyen de recepteurs peptidiques intestinaux vasoactifs | |
North et al. | All three vasopressin receptor sub-types are expressed by small-cell carcinoma | |
Gerdes et al. | Signal-mediated sorting of chromogranins to secretory granules | |
Karacay et al. | Expression and fine mapping of murine vasoactive intestinal peptide receptor 1 | |
EP1437592A1 (fr) | Nouvelle methode de criblage au moyen du recepteur de prokineticine | |
Alokail et al. | Quantitative comparison of PTH1R in breast cancer MCF7 and osteosarcoma SaOS‐2 cell lines | |
Clauser et al. | The human insulin receptor cDNA: a new tool to study the function of this receptor | |
JPH1118782A (ja) | 新規なヒト・ニューロテンシン・レセプター2型およびそのスプライス変種 | |
WO2001036446A2 (fr) | Traitement de maladies | |
US20030171293A1 (en) | Neuropeptide receptor and uses thereof | |
JPH11137273A (ja) | 新規化合物 | |
CHOREV et al. | Interactions of Parathyroid Hormone and Parathyroid Hormone-Related |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): CA JP MX |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
122 | Ep: pct application non-entry in european phase | ||
NENP | Non-entry into the national phase |
Ref country code: CA |