US20060228778A1 - Novel vitamin D receptor related polypeptides, nucleic acid sequence encoding the same and uses thereof - Google Patents
Novel vitamin D receptor related polypeptides, nucleic acid sequence encoding the same and uses thereof Download PDFInfo
- Publication number
- US20060228778A1 US20060228778A1 US11/369,263 US36926306A US2006228778A1 US 20060228778 A1 US20060228778 A1 US 20060228778A1 US 36926306 A US36926306 A US 36926306A US 2006228778 A1 US2006228778 A1 US 2006228778A1
- Authority
- US
- United States
- Prior art keywords
- vdrr
- nucleic acid
- acid sequence
- polypeptide
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 85
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 85
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 84
- 150000007523 nucleic acids Chemical group 0.000 title claims abstract description 52
- 102000009310 vitamin D receptors Human genes 0.000 title claims abstract description 29
- 108050000156 vitamin D receptors Proteins 0.000 title claims abstract description 29
- 108091028043 Nucleic acid sequence Proteins 0.000 title claims description 16
- 238000000034 method Methods 0.000 claims abstract description 32
- 239000000126 substance Substances 0.000 claims abstract description 27
- 239000003814 drug Substances 0.000 claims abstract description 14
- 230000014509 gene expression Effects 0.000 claims abstract description 14
- 230000019491 signal transduction Effects 0.000 claims abstract description 14
- 239000003446 ligand Substances 0.000 claims abstract description 13
- 230000002062 proliferating effect Effects 0.000 claims abstract description 12
- 230000004968 inflammatory condition Effects 0.000 claims abstract description 11
- 230000002503 metabolic effect Effects 0.000 claims abstract description 11
- 239000013604 expression vector Substances 0.000 claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 claims abstract description 10
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 46
- 102000039446 nucleic acids Human genes 0.000 claims description 33
- 108020004707 nucleic acids Proteins 0.000 claims description 33
- 108020001756 ligand binding domains Proteins 0.000 claims description 28
- 241000282414 Homo sapiens Species 0.000 claims description 26
- 239000002299 complementary DNA Substances 0.000 claims description 22
- 235000001014 amino acid Nutrition 0.000 claims description 13
- 150000001413 amino acids Chemical class 0.000 claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 13
- 230000004568 DNA-binding Effects 0.000 claims description 8
- 239000000523 sample Substances 0.000 claims description 7
- 238000003556 assay Methods 0.000 claims description 6
- 239000002773 nucleotide Substances 0.000 claims description 6
- 125000003729 nucleotide group Chemical group 0.000 claims description 6
- 108020004711 Nucleic Acid Probes Proteins 0.000 claims description 5
- 239000005557 antagonist Substances 0.000 claims description 5
- 238000000338 in vitro Methods 0.000 claims description 5
- 239000002853 nucleic acid probe Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 239000013598 vector Substances 0.000 claims description 5
- 208000031226 Hyperlipidaemia Diseases 0.000 claims description 4
- 208000008589 Obesity Diseases 0.000 claims description 4
- 206010012601 diabetes mellitus Diseases 0.000 claims description 4
- 238000009396 hybridization Methods 0.000 claims description 4
- 235000020824 obesity Nutrition 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
- 108700028369 Alleles Proteins 0.000 claims description 3
- 108090001030 Lipoproteins Proteins 0.000 claims description 3
- 102000004895 Lipoproteins Human genes 0.000 claims description 3
- 241000124008 Mammalia Species 0.000 claims description 3
- 206010028980 Neoplasm Diseases 0.000 claims description 3
- 108020004511 Recombinant DNA Proteins 0.000 claims description 3
- 239000000556 agonist Substances 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 3
- 235000018102 proteins Nutrition 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 230000001131 transforming effect Effects 0.000 claims description 3
- 208000035150 Hypercholesterolemia Diseases 0.000 claims description 2
- 201000002980 Hyperparathyroidism Diseases 0.000 claims description 2
- 208000001132 Osteoporosis Diseases 0.000 claims description 2
- 208000022531 anorexia Diseases 0.000 claims description 2
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 2
- 206010061428 decreased appetite Diseases 0.000 claims description 2
- 230000007547 defect Effects 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 208000020346 hyperlipoproteinemia Diseases 0.000 claims description 2
- 230000003463 hyperproliferative effect Effects 0.000 claims description 2
- 238000001727 in vivo Methods 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 238000000159 protein binding assay Methods 0.000 claims description 2
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 2
- 208000017520 skin disease Diseases 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims 1
- 239000007924 injection Substances 0.000 claims 1
- 108020005497 Nuclear hormone receptor Proteins 0.000 abstract description 28
- 108020004017 nuclear receptors Proteins 0.000 abstract description 27
- 238000009472 formulation Methods 0.000 abstract description 4
- 239000000203 mixture Substances 0.000 abstract description 4
- 230000001105 regulatory effect Effects 0.000 abstract description 4
- 102000006255 nuclear receptors Human genes 0.000 abstract 1
- 102000007399 Nuclear hormone receptor Human genes 0.000 description 27
- 210000004027 cell Anatomy 0.000 description 24
- 102000005962 receptors Human genes 0.000 description 17
- 108020003175 receptors Proteins 0.000 description 17
- 210000001519 tissue Anatomy 0.000 description 11
- 239000012190 activator Substances 0.000 description 10
- 102000034527 Retinoid X Receptors Human genes 0.000 description 9
- 108010038912 Retinoid X Receptors Proteins 0.000 description 9
- VSBHRRMYCDQLJF-ZDNYCOCVSA-N pregnenolone 16alpha-carbonitrile Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1C[C@@H](C#N)[C@H](C(=O)C)[C@@]1(C)CC2 VSBHRRMYCDQLJF-ZDNYCOCVSA-N 0.000 description 9
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 7
- 210000001072 colon Anatomy 0.000 description 7
- 210000004185 liver Anatomy 0.000 description 7
- 102000004164 orphan nuclear receptors Human genes 0.000 description 7
- 108090000629 orphan nuclear receptors Proteins 0.000 description 7
- 210000000813 small intestine Anatomy 0.000 description 7
- 239000011647 vitamin D3 Substances 0.000 description 7
- 229940021056 vitamin d3 Drugs 0.000 description 7
- 108020004635 Complementary DNA Proteins 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 6
- 235000005282 vitamin D3 Nutrition 0.000 description 6
- 230000004913 activation Effects 0.000 description 5
- XMRPGKVKISIQBV-UHFFFAOYSA-N (+-)-5- Pregnane-3,20-dione Natural products C1CC2CC(=O)CCC2(C)C2C1C1CCC(C(=O)C)C1(C)CC2 XMRPGKVKISIQBV-UHFFFAOYSA-N 0.000 description 4
- DUHUCHOQIDJXAT-CSXWOMMHSA-N (3r,5r,8s,9s,10s,13s,14s,17s)-17-acetyl-3-hydroxy-10,13-dimethyl-1,2,3,4,5,6,7,8,9,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-11-one Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](C(=O)C)[C@@]2(C)CC1=O DUHUCHOQIDJXAT-CSXWOMMHSA-N 0.000 description 4
- ORNBQBCIOKFOEO-YQUGOWONSA-N Pregnenolone Natural products O=C(C)[C@@H]1[C@@]2(C)[C@H]([C@H]3[C@@H]([C@]4(C)C(=CC3)C[C@@H](O)CC4)CC2)CC1 ORNBQBCIOKFOEO-YQUGOWONSA-N 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 230000027455 binding Effects 0.000 description 4
- 229960003957 dexamethasone Drugs 0.000 description 4
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 4
- -1 fatty acid esters Chemical class 0.000 description 4
- 108020001507 fusion proteins Proteins 0.000 description 4
- ORNBQBCIOKFOEO-QGVNFLHTSA-N pregnenolone Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 ORNBQBCIOKFOEO-QGVNFLHTSA-N 0.000 description 4
- 229960000249 pregnenolone Drugs 0.000 description 4
- 150000003131 pregnenolone derivatives Chemical class 0.000 description 4
- 150000003132 pregnenolones Chemical class 0.000 description 4
- 102000003702 retinoic acid receptors Human genes 0.000 description 4
- 108090000064 retinoic acid receptors Proteins 0.000 description 4
- XMRPGKVKISIQBV-XWOJZHJZSA-N 5beta-pregnane-3,20-dione Chemical compound C([C@H]1CC2)C(=O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](C(=O)C)[C@@]2(C)CC1 XMRPGKVKISIQBV-XWOJZHJZSA-N 0.000 description 3
- 238000001712 DNA sequencing Methods 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 108091060211 Expressed sequence tag Proteins 0.000 description 3
- 108060001084 Luciferase Proteins 0.000 description 3
- 239000005089 Luciferase Substances 0.000 description 3
- 102000003728 Peroxisome Proliferator-Activated Receptors Human genes 0.000 description 3
- 108090000029 Peroxisome Proliferator-Activated Receptors Proteins 0.000 description 3
- 108010029485 Protein Isoforms Proteins 0.000 description 3
- 102000001708 Protein Isoforms Human genes 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 230000000708 anti-progestin effect Effects 0.000 description 3
- 239000003418 antiprogestin Substances 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 102000037865 fusion proteins Human genes 0.000 description 3
- 238000003146 transient transfection Methods 0.000 description 3
- YWYQTGBBEZQBGO-OQOYYRQQSA-N (3r,5r,8r,9s,10s,13s,14s,17s)-17-[(1r)-1-hydroxyethyl]-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-3-ol Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](O)C)[C@@]2(C)CC1 YWYQTGBBEZQBGO-OQOYYRQQSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- GMRQFYUYWCNGIN-UHFFFAOYSA-N 1,25-Dihydroxy-vitamin D3' Natural products C1CCC2(C)C(C(CCCC(C)(C)O)C)CCC2C1=CC=C1CC(O)CC(O)C1=C GMRQFYUYWCNGIN-UHFFFAOYSA-N 0.000 description 2
- 150000000631 5β-pregnanes Chemical class 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 102000004328 Cytochrome P-450 CYP3A Human genes 0.000 description 2
- 108010081668 Cytochrome P-450 CYP3A Proteins 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 229940123037 Glucocorticoid antagonist Drugs 0.000 description 2
- 101000640876 Homo sapiens Retinoic acid receptor RXR-beta Proteins 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-M Methanesulfonate Chemical compound CS([O-])(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-M 0.000 description 2
- 108700005081 Overlapping Genes Proteins 0.000 description 2
- 108700008625 Reporter Genes Proteins 0.000 description 2
- 102100034253 Retinoic acid receptor RXR-beta Human genes 0.000 description 2
- AUYYCJSJGJYCDS-LBPRGKRZSA-N Thyrolar Chemical class IC1=CC(C[C@H](N)C(O)=O)=CC(I)=C1OC1=CC=C(O)C(I)=C1 AUYYCJSJGJYCDS-LBPRGKRZSA-N 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- YWYQTGBBEZQBGO-UHFFFAOYSA-N UC1011 Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(O)C)C1(C)CC2 YWYQTGBBEZQBGO-UHFFFAOYSA-N 0.000 description 2
- 241000269368 Xenopus laevis Species 0.000 description 2
- 235000020964 calcitriol Nutrition 0.000 description 2
- 239000011612 calcitriol Substances 0.000 description 2
- GMRQFYUYWCNGIN-NKMMMXOESA-N calcitriol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CCCC(C)(C)O)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C GMRQFYUYWCNGIN-NKMMMXOESA-N 0.000 description 2
- 230000024245 cell differentiation Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 239000013613 expression plasmid Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000003635 glucocorticoid antagonist Substances 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 150000003431 steroids Chemical class 0.000 description 2
- 239000005495 thyroid hormone Substances 0.000 description 2
- 229940036555 thyroid hormone Drugs 0.000 description 2
- HEGJUOCNFOFQSY-ALEFBPAYSA-N (5R,8S,9S,10R,13S,14S)-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-4-ol Chemical class [C@@H]12CCC[C@@]1(C)CC[C@H]1[C@H]2CC[C@H]2C(CCC[C@]12C)O HEGJUOCNFOFQSY-ALEFBPAYSA-N 0.000 description 1
- AHRWWYGWQKBKBF-PCHYHQBZSA-N (5r,8s,9s,10s,13s,14s,17s)-17-acetyl-10,13-dimethyl-2,4,5,6,7,8,9,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthrene-3,11-dione Chemical compound C([C@H]1CC2)C(=O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](C(=O)C)[C@@]2(C)CC1=O AHRWWYGWQKBKBF-PCHYHQBZSA-N 0.000 description 1
- GEKLXXUPTBLQJU-ARHWZJKSSA-N (5s,8s,9s,10s,13s,14s)-10,13-dimethylspiro[2,4,5,6,7,8,9,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthrene-3,2'-3h-1,3-benzothiazole]-11-one Chemical compound N1C2=CC=CC=C2SC11C[C@@H]2CC[C@H]3[C@@H]4CCC[C@@]4(C)CC(=O)[C@@H]3[C@@]2(C)CC1 GEKLXXUPTBLQJU-ARHWZJKSSA-N 0.000 description 1
- XZHKZJXZRWFLCJ-RKFIYKRSSA-N (8R,9S,10S,13R,14S,17R)-17-ethyl-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,17-dodecahydro-1H-cyclopenta[a]phenanthrene-15,16-dione Chemical class [C@@H]12C(C([C@H](CC)[C@@]1(C)CC[C@H]1[C@H]2CCC2CCCC[C@]12C)=O)=O XZHKZJXZRWFLCJ-RKFIYKRSSA-N 0.000 description 1
- PXVQJWLYGCMTTP-RKFIYKRSSA-N (8R,9S,10S,13S,14S,17S)-17-acetyl-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,17-dodecahydro-1H-cyclopenta[a]phenanthrene-15,16-dione Chemical class [C@@H]12C(C([C@H](C(C)=O)[C@@]1(C)CC[C@H]1[C@H]2CCC2CCCC[C@]12C)=O)=O PXVQJWLYGCMTTP-RKFIYKRSSA-N 0.000 description 1
- HZINQBCDAHFWOC-PTPDVJPESA-N (8r,9r,10s,13s,14s)-13-methyl-1,2,3,4,5,6,7,8,9,10,11,12,14,15,16,17-hexadecahydrocyclopenta[a]phenanthren-3-ol Chemical compound C1CC2CC(O)CC[C@@H]2[C@@H]2[C@@H]1[C@@H]1CCC[C@@]1(C)CC2 HZINQBCDAHFWOC-PTPDVJPESA-N 0.000 description 1
- IJFGLPBXLYRFEJ-INGDRISUSA-N (8r,9s,10s,13r,14s,17s)-17-ethyl-10,13-dimethyl-1,2,3,4,5,6,7,8,9,11,12,14,16,17-tetradecahydrocyclopenta[a]phenanthren-15-one Chemical class C1CC2CCCC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1C(=O)C[C@H](CC)[C@@]1(C)CC2 IJFGLPBXLYRFEJ-INGDRISUSA-N 0.000 description 1
- XCESQNFRBAHSGO-WRJHFWDFSA-N (8s,10s,13s,14s,17s)-17-acetyl-10,13-dimethyl-6,7,8,12,14,15,16,17-octahydrocyclopenta[a]phenanthren-3-one Chemical compound O=C1C=C[C@]2(C)C3=CC[C@]4(C)[C@@H](C(=O)C)CC[C@H]4[C@@H]3CCC2=C1 XCESQNFRBAHSGO-WRJHFWDFSA-N 0.000 description 1
- QLZSNRDVVCGCPO-HNRKPFPWSA-N (8s,9s,10s,13r,14s,17s)-17-ethyl-10,13-dimethylspiro[1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthrene-3,2'-1,3-thiazolidine]-4'-carboxylic acid Chemical compound C([C@H]1[C@@H]2CC[C@@H]([C@]2(CC[C@@H]1[C@@]1(C)CC2)C)CC)CC1CC12NC(C(O)=O)CS1 QLZSNRDVVCGCPO-HNRKPFPWSA-N 0.000 description 1
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 1
- FPMXPTIRDWHULR-UHFFFAOYSA-N 2-pyridin-1-ium-1-ylacetate;hydrochloride Chemical compound [Cl-].OC(=O)C[N+]1=CC=CC=C1 FPMXPTIRDWHULR-UHFFFAOYSA-N 0.000 description 1
- XMRPGKVKISIQBV-BJMCWZGWSA-N 5alpha-pregnane-3,20-dione Chemical compound C([C@@H]1CC2)C(=O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](C(=O)C)[C@@]2(C)CC1 XMRPGKVKISIQBV-BJMCWZGWSA-N 0.000 description 1
- SHGAZHPCJJPHSC-ZVCIMWCZSA-N 9-cis-retinoic acid Chemical compound OC(=O)/C=C(\C)/C=C/C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-ZVCIMWCZSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- PQSUYGKTWSAVDQ-UHFFFAOYSA-N Aldosterone Natural products C1CC2C3CCC(C(=O)CO)C3(C=O)CC(O)C2C2(C)C1=CC(=O)CC2 PQSUYGKTWSAVDQ-UHFFFAOYSA-N 0.000 description 1
- PQSUYGKTWSAVDQ-ZVIOFETBSA-N Aldosterone Chemical compound C([C@@]1([C@@H](C(=O)CO)CC[C@H]1[C@@H]1CC2)C=O)[C@H](O)[C@@H]1[C@]1(C)C2=CC(=O)CC1 PQSUYGKTWSAVDQ-ZVIOFETBSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 1
- 229930105110 Cyclosporin A Natural products 0.000 description 1
- 108010036949 Cyclosporine Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 1
- 102100039556 Galectin-4 Human genes 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101000608765 Homo sapiens Galectin-4 Proteins 0.000 description 1
- 101000603877 Homo sapiens Nuclear receptor subfamily 1 group I member 2 Proteins 0.000 description 1
- 101001126471 Homo sapiens Plectin Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 238000000636 Northern blotting Methods 0.000 description 1
- 102100038494 Nuclear receptor subfamily 1 group I member 2 Human genes 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 108091008731 RAR-related orphan receptors α Proteins 0.000 description 1
- 108091027981 Response element Proteins 0.000 description 1
- 102100023606 Retinoic acid receptor alpha Human genes 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 238000012896 Statistical algorithm Methods 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 229960002478 aldosterone Drugs 0.000 description 1
- 229960001445 alitretinoin Drugs 0.000 description 1
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 1
- 230000037354 amino acid metabolism Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- CBMYJHIOYJEBSB-CAHXEBCQSA-N androstane-3,17-diol Chemical class C1C(O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)O)[C@@H]4[C@@H]3CCC21 CBMYJHIOYJEBSB-CAHXEBCQSA-N 0.000 description 1
- 150000001441 androstanes Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 150000001491 aromatic compounds Chemical class 0.000 description 1
- 238000000376 autoradiography Methods 0.000 description 1
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000023852 carbohydrate metabolic process Effects 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 230000023715 cellular developmental process Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 150000001841 cholesterols Chemical class 0.000 description 1
- 229960001265 ciclosporin Drugs 0.000 description 1
- 238000011260 co-administration Methods 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 239000008139 complexing agent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229930182912 cyclosporin Natural products 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 208000037765 diseases and disorders Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 125000002250 eicosanoid group Chemical group 0.000 description 1
- 229960005309 estradiol Drugs 0.000 description 1
- 229930182833 estradiol Natural products 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 238000005734 heterodimerization reaction Methods 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 239000004030 hiv protease inhibitor Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000008463 key metabolic pathway Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 102000004311 liver X receptors Human genes 0.000 description 1
- 108090000865 liver X receptors Proteins 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 150000003127 pregnanediols Chemical class 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 229940077150 progesterone and estrogen Drugs 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 229940127293 prostanoid Drugs 0.000 description 1
- 150000003814 prostanoids Chemical class 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 230000009711 regulatory function Effects 0.000 description 1
- 238000003571 reporter gene assay Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 210000001995 reticulocyte Anatomy 0.000 description 1
- 229930002330 retinoic acid Natural products 0.000 description 1
- 108091008726 retinoic acid receptors α Proteins 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 108020001568 subdomains Proteins 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 229940037128 systemic glucocorticoids Drugs 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- 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
- C07K14/721—Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
Abstract
The present invention relates to novel vitamin D receptor related (VDRR) polypeptides, and formulations containing the same. Nucleic acid sequences encoding the VDRR polypeptides, expression vectors containing such sequences and host cells transformed with such expression vectors are also disclosed, as are methods for the expression of the novel VDRR polypeptides of the invention. The invention further relates to VDRR polypeptides for use as medicaments, and use of substances affecting VDRR signal transduction for the manufacture of medicaments for metabolic, proliferative or inflammatory conditions. The present invention also relates to methods for identifying clones encoding a VDRR polypeptide, methods for identifying ligands to a VDRR and methods for identifying substances for treatment of conditions affected by a VDRR polypeptide. More specifically, the novel VDRR polypeptide can be the polypeptide designated VDRRγ, which may be regulated by any small chemical molecule similar in structure to known ligands for nuclear receptors.
Description
- The present invention relates to novel vitamin D receptor related (VDRR) polypeptides. Nucleic acid sequences encoding the same, expression vectors containing such sequences and host cells transformed with such expression vectors are also disclosed, as are methods for the expression of the novel VDRR polypeptides of the invention, and uses thereof.
- Nuclear hormone receptors is a large group of conditionally regulated transcription factors. These receptors are activated and regulate target gene expression in response to binding a variety of small chemical molecules (ligands) including steroids, vitamin D3, retinoids, eicosanoides (prostanoids), thyroid hormone and cholesterol derivatives.
- A growing number of structurally related receptors have been identified for which no ligands yet have been identified. This group of receptors is referred to as orphan nuclear receptors (ONRs). A review of the ONRs can be found in Enmark et al, Mol. Endo., vol. 10, No. 11 (1996) pp. 1293-1307, which is hereby incorporated by reference. The pivotal importance of a number of ONRs for processes such as metabolic homeostasis, cell differentiation and development have been demonstrated both by biochemical and genetic techniques. In addition, several ONRs have also been implicated as key factors in a variety of common diseases and disorders such as diabetes, obesity, inflammatory conditions and proliferative diseases.
- Based on these findings it is generally believed that novel ONRs are going to become potential drug targets for therapeutic invention of common diseases. Thus, it is of great importance to identify such receptors.
- The present invention relates to novel vitamin D receptor related (VDRR) polypeptides, and formulations containing the same. Nucleic acid sequences encoding the VDRR polypeptides, expression vectors containing such sequences and host cells transformed with such expression vectors are also disclosed, as are methods for the expression of the novel VDRR polypeptides of the invention. The invention further relates to VDRR polypeptides for use as medicaments, and use of substances affecting VDRR signal transduction for the manufacture of medicaments for treating metabolic, proliferative or inflammatory conditions. The present invention also relates to methods for identifying clones encoding a VDRR polypeptide, methods for identifying ligands to a VDRR and methods for identifying substances for treatment of conditions affected by a VDRR polypeptide. More specifically, the novel VDRR polypeptide can be the polypeptide designated VDRRγ, which may be regulated by any small chemical molecule similar in structure to known ligands for nuclear receptors.
-
FIG. 1 —The cDNA sequence encoding the novel nuclear receptor polypeptide vitamin D receptor related gamma (VDRRg) is shown. -
FIG. 2 —Evolutionary neighbor-joining tree for VDRRg as given by DBD-HMM alignment. -
FIG. 3 —Evolutionary neighbor-joining tree for VDRRg as given by LBD-HMM alignment. -
FIG. 4 —The deduced amino acid sequence of VDRRg is shown. -
FIG. 5 —Expression of VDRRg in adult human tissues. The numbers on the right hand side, refer to kilobasepairs of the mRNA. -
FIG. 6 —Vitamin D3 transactivate a GAL4-DBD/VDR-LBD fusion protein but not a GAL4-DBD/VDRRγ-LBD fusion protein in transient transfections of CV-1 cells. The number on the left hand side refer to relative luciferase activity of the GAL4-luciferase reporter gene. -
FIG. 7 —The cDNA sequence encoding VDRRg-2 with an alternatively spliced 5-end compared to VDRRg is shown. -
FIG. 8 —The deduced amino acid sequence of VDRRg-2 is shown. -
FIG. 9 —Heterodimerization of VDRRg with a retinoid X receptor (RXR) is shown. -
FIG. 10 —The effect of pregnenolone derivatives as activators of VDRRg are shown. -
FIG. 11 —The effect of pregnenolone 16α-carbonitrile (PCN), dexamethasone and an antiprogestin (RU486) as activators of VDRRg are shown. -
FIG. 12 —Percent similarity between the new genes VDRRg-1 and VDRRg-2 and the known genes XOR-6. HVDR, CAR-1 and CAR-2. -
FIG. 13 —Percent identity between the new genes VDRRg-1 and VDRRg-2 and the known genes XOR-6. HVDR, CAR-1 and CAR-2. - The objects above are met by the present invention, which relates to a mammalian, preferably human, isolated or recombinant nucleic acid comprising a contiguous nucleic acid sequence encoding a vitamin D receptor related (VDRR) polypeptide. The VDRR polypeptide is suitably origin.
- In preferred embodiments of the present invention, the nucleic acid encoding the VDRR polypeptide contains a DNA-binding domain (DBD) comprising about 77 amino acids with 9 cysteine residues. The DBD is further characterized by the following amino acid sequence similarity relative to the DBDs of human Vitamin D Receptor (hVDR) and Orphan
Nuclear Receptor 1 isolated from Xenopus laevis (xONR1=XOR-6), respectively: - (i) at least about 60% amino acid sequence similarity with thc DBD of hVDR; and
- (ii) at least about 65% amino acid sequence similarity with the DBD of xONR1.
More particularly, the amino acid sequence similarity relative to the DBDs of hVDR and xONR1, respectively is - (i) about 65% amino acid sequence similarity with the DBD of hVDR; and
- (ii) about 71% amino acid sequence similarity with the DBD of xONR1.
- In preferred embodiments of the present invention, the nucleic acid encoding the VDRR polypeptide contains a ligand-binding domain (LBD) characterized by the following amino acid sequence similarity, relative to the LBDs of hVDR and xONR1, respectively:
- (i) at least about 30% amino acid sequence similarity with the LBD of hVDR, suitably at least 35% amino acid sequence similarity with the LBD of hVDR; and
- (ii) at least about 40% amino acid sequence similarity with the LBD of xONR1, suitably at least 45% amino acid sequence similarity with the LBD of xONR1.
More particularly, the amino acid sequence similarity relative to the LBDs of hVDR and xONR1, respectively is - (i) about 42% amino acid sequence similarity with the LBD of hVDR; and
- (ii) about 54% amino acid sequence similarity with the LBD of xONR1.
“amino acid sequence similarity” refers to: 100×Consensus Lenght divided by Consencus Length+Mismatsches+Gaps.
The term amino acid sequence identity can also be used. Amino acid sequence identity is calculated by comparing the absolute amino acid residue identity. InFIG. 13 the amino acid sequence identity between the new genes VDRRg-1 and VDRRg-2 and the known genes are shown. - In particularly preferred embodiments, the nucleic acid sequences of the present invention are substatially the same as those given in
FIG. 1 orFIG. 7 , the same or alleles thereof. - The present invention also relates to a nucleic acid probe for the detection of a nucleic acid sequence encoding a VDRR polypeptide in a sample. Suitably, the probe comprises at least 14 contiguous nucleotides, and preferably at least 28 contiguous nucleotides, of the nucleic acid sequences given in
FIG. 1 orFIG. 7 . The nucleic acid probe can be used in a method for identifying clones encoding a VDRR polypeptide, wherein the method comprises screening a genomic or cDNA library with the probe under low stringency hybridization conditions, and identifying those clones which display a substantial degree of hybridization to said probe. - The present invention further relates to an isolated or recombinant VDRR polypeptide. The polypeptide can be full-length, at which the sequence of amino acids is identical to the corresponding sequence found in mammals in general, and in human beings in particular. In the present invention, the polypeptide can also be a truncated, extended or mutated form of the full-length polypeptide. Truncated and extended forms relate to VDRR polypeptides where one or more amino acids are missing or have been added, respectively, at the N terminal end of the polypeptide chain. Mutated forms relate to VDRR polypep-tides where one or more amino acid has been substituted by another amino acid. Suitably, the isolated or recombinant VDRR polypeptide exhibits the amino acid sequences given in
FIG. 4 orFIG. 8 . - The N-terminal sequence of the present nucleic acids encoding VDRR polypeptides, as well as the amino acid sequence of the present VDRR polypeptides, may vary. Thus, various N-terminal isoforms are envisaged, e.g. any of α1, α2, β1, β2, β3, β4, γ1 or γ2 as disclosed in FIG. 7B of Transcription Factors 3: nuclear receptors, Protein Profile, vol. 2, issue 11 (1995), pp. 1173-1235. This review of nuclear receptors generally is hereby incorporated by reference. More specifically, Vitamin D receptors and related orphans, e.g. ONR1, are discussed at p. 1191-1992.
- The present invention further relates to pharmaceutical formulations comprising an isolated or recombinant VDRR polypeptide, and one or more therapeutically acceptable excipients. Examples of excipients that can be used are carbohydrates, e.g. monosaccharides, disaccharides and sugar alcohols, such as saccharose and sorbitol. Further examples include amino acids, e.g. histidine and arginine, surfactants, e.g. polyoxyethylene sorbitan fatty acid esters, inorganic salts, e.g. sodium chloride and calcium chloride, and complexing agents, e.g. EDTA and citric acid.
- The present formulation can be in the form of an aqueous solution ready-for-use, or dried, particularly lyophilized. In the latter case, the formulation is reconstituted with a liquid, e.g. sterile water or saline, before use.
- The present invention further relates to an expression vector comprising an isolated or recombinant nucleic acid, the nucleic acid comprising a contiguous nucleic acid sequence encoding a Vitamin D receptor related (VDRR) polypeptide. The invention also relates to a cell containing such an expression vector.
- The present invention further relates to a cell containing the claimed nucleic acid, the nucleic acid comprising a contiguous nucleic acid sequence encoding a Vitamin D receptor related (VDRR) polypeptide.
- The present invention further relates to a process for recombinant production of a VDRR polypeptide, by expressing the claimed isolated or recombinant contiguous nucleic acid sequence encoding a Vitamin D receptor related (VDRR) polypeptide in a suitable host cell, preferably an eukaryotic cell.
- The present invention further relates to method for identifying a ligand to a VDRR, e.g. by a cell-based reporter assay, transgenic-animal reporter assay or in vitro-binding assay. It also relates to a method for identifying a substance for treatment of a condition affected by a VDRR polypeptide, comprising screening for an agonist or an antagonist of VDRR polypeptide signal transduction to be used for treating metabolic, proliferative or inflammatory conditions.
- The present invention further relates to a VDRR polypeptide for use as a medicament, as well as use of a substance affecting VDRR signal transduction for the manufac-ture of a medicament for treating metabolic, proliferative or inflammatory conditions.
- More particularly, the present invention can be used for the manufacture of medicaments for treating obesity, diabetes, anorexia, lipoprotein defects, hyperlipidemia, hypercholeste-remia or hyperlipoproteinemia. The present invention can be used also for the manufacture of medicaments for treating osteoporosis, rheumatoid arthritis, benign and malign tumors, hyperproliferative skin disorders or hyperparathyroidism.
- The present invention further relates to a method for treating metabolic, proliferative or inflammatory conditions by introducing into a mammal a nucleic acid vector encoding for expression of a VDRR polypeptide. The nucleic acid vector is capable of transforming a cell in vivo and expressing said polypeptide in said transformed cell.
- The present invention further relates to a method for treatment of a metabolic, proliferative or inflammatory condition by administration of a therapeutically effective amount of a substance affecting VDRR signal transduction, specifically a VDRR polypeptide.
- In the present invention, the term “isolated” in connection with VDRR polypeptides or nucleic acids encoding the same, relates to nucleic acids or polypeptides that have been isolated from a natural source, e.g. the liver, small intestine or colon of a human being. The isolated VDRR polypeptides or nucleic acids of the present invention are unique in the sense that they are not found in a pure or separated form in nature. Use of the term “isolated” indicates that a naturally occurring sequence has been removed from its normal cellular environment. Thus, the sequence may be in a cell-free environment or in a different cellular environment. The term does not imply that the sequence is the only nucleic acid or amino acid sequence present, but that it is the predominant nucleic acid or amino acid sequence present. Furthermore, the nucleic acid or polypeptide should be essentially free of non-amino acid or non-nucleic acid material naturally associated with the respective product. In this context, essentially free relates to more than 80%, suitably more than 90%, and preferably more than 95% purity.
- The term “substantially the same” when referring to the nucleic acid sequences in
FIG. 1 orFIG. 7 and when referring to the amino acid sequence inFIG. 4 orFIG. 8 means that they are derived from the sequences given in the figures and have the same function as those. - The inventors of the present invention, have surprisingly isolated a novel nucleic acid sequence, and a polypeptide encoded by said nucleic acid sequence. Thus, a novel cDNA encoding a polypeptide designated VDRRγ has been cloned and characterized. This polypeptide is, based on amino acid sequence similarity, a novel member of the nuclear (hormone) receptor supergene family. Hidden Markov Models (HMMs) in combination with phylogenetic analysis such as neighbor-joining tree methods and other statistical algorithms shows that VDRRγ belong to a sub-family of vitamin D receptors (VDRs) and a VDR-like receptor from Xenopus laevis designated xONR1 (see Smith et al., Nucl. Acids Res., 22 (1994), No. 1, pp. 66-71) or XOR-6 as in WO96/22390. The VDRRγ, therefore, is one member of a family of Vitamin D receptor related (VDRR) polypeptides.
- The degree of amino acid similarity in the DBD and LBD of VDRRg as compared to the most closely related receptors XOR-6, hVDR and CAR (see WO 93/17041) is similar to the relationship between other distinct, but related nuclear receptors. (See
FIG. 12 ). The thyroid hormone (TRb) and retinoic acid receptor (RARb) are approximately 60% and 40% identical at the amino acid level in the DBD and LBD, respectively. By comparison, the closely related but unique genes encoding human RARa and RARb nuclear receptors are 97% and 82% identical in the DBD and LBD, respectively. - As recognized by those skilled in the art of nuclear receptors, the DBD displays the highest degree of conservation (amino acid identity) both between different nuclear receptors (paralogous) and between identical receptors from different species (orthologues). The two “zink-fingers” in the DBD are generated by two evolutionary conserved amino acid motifs Cys-X2-Cys-X13-Cys-X2-Cys (amino-terminal or first zink-finger) and Cys-Xn-Cys-X9-Cys-X2-Cys (carboxy-terminal or second zink-finger) in which two pairs of cysteins chelate on zink ion. The vast majority of nuclear receptors have five amino acid residues between the firs two Cys residues in the second zink-finger (Cys-X5-Cys-X9-Cys-X2-Cys) see Gronemeyer and Laudet (
Protein Profile 1995, 2, issue 11) for details. The today only known exception to this role are the PPARs which have three amino acid (Cys-X3-Cys-X9-Cys-X2-Cys) residues and the TLL group of receptors which have seven (Cys-X7-Cys-X9-Cys-X2-Cys). Thus another feature which is characteristic of the novel VDRRg polypeptide described herein is that the number of amino acid residues in this part of the DBD is six (Cys-X6-Cys-X9-Cys-X2-Cys) as shown inFIGS. 4 and 8 . Today, the only other nuclear receptor like sequences found in the TREMBLE data base with the same number of amino acid residues between the two cys residues are two sequences (Q20097 and Q18155) from the worm C. elegans (Q20097 and Q18155). However, the entire DBD of these putative C. elegans nuclear receptors are only distantly related to the DBD of VDRRg. Taken together, the comparison of the DBD and LBD of the nuclear receptor VDRRg described herein (SeeFIG. 12 ), clearly demonstrate that this receptor is a novel member of the nuclear receptor super-gene family which is distinct from other known nuclear receptors that are most closely related to the VDRRg including ONR-1 (in Smith et al., 1994, Nucleic Acids Res., 22, pp 66-71) or XOR-6 (in WO 96/22390), hVDR and CAR (WO 93/17041). - This finding, in combination with the highly restricted expression pattern we observe for human VDRRγ (liver, small intestine and mucosa of colon) and in analogy to other nuclear receptors exhibiting a tissue specific expression pattern such as the peroxisome pro-liferator-activated receptors (PPARs)—suggest that VDRRγ performs important physiolo-gical functions in liver, small intestine and colon. Accordingly. VDRRγ is likely to be an important sensor of key metabolic pathways affecting lipid, carbohydrate or amino acid metabolism/homeostasis. In addition, the highly selective tissue specific expression pattern suggest that VDRRγ may participate in cellular differentiation and development of these tissues.
- An additional human VDRRγ cDNA with an alternatively spliced 5′-end has been identified (see
FIG. 7 ). The VDRRγ cDNAs are thus able to encode at least one alternative N-terminal variant (FIG. 8 ) in addition to the VDRRγ polypeptide shown inFIG. 4 . In analogy to other members of the nuclear receptor supergene family such as RORα and RARα these N-terminal isoforms of VDRRγ may specify different functions including DNA-binding specificity and/or promoter specific activation (Gronemeyer and Laudet, 1995). - In the present specification, the term VDRRγ relates to the various polypeptides corresponding to the differentially spliced VDRRγ cDNAs including VDRRγ-1 and VDRRγ-2. However, when reference is made to
FIG. 1 andFIG. 4 , VDRRγ cDNA and VDRRγ relates specifically to VDRRγ-1 cDNA and VDRRγ-1, respectively. In the same way, when reference is made toFIG. 7 andFIG. 8 , VDRRγ cDNA and VDRRγ relates specifically to VDRRγ-2 cDNA and VDRRγ-2, respectively. - In contrast to the VDRRγ-2 cDNA, the VDRRγ-1 cDNA does not contain a classical AUG initiation codon but instead may initiate at an alternative CUG codon. This putative non-AUG start site is located in a favorable sequence context for efficient initiation from alternative start sites and is in frame with the entire open reading frame and preceded by a stop codon.
- Taken together, the VDRRs in general, and more specifically the VDRRγ, may be important in
-
- 1) metabolic diseases such as obesity, diabetes (type I and II), lipoprotein disorders,
- 2) proliferative conditions such as tumors (benign and malignant) of the small intestine and colon,
- 3) ulcero-inflammatory diseases of small intestine and colon such as Crohn's disease and ulcerative colitis, and
- 4) congenital anomalies of small intestine and colon.
- The high amino acid sequence identity of VDRRγ with the VDR both in the DNA-binding domain (DBD) and ligand-binding domain (LBD) indicate that these two receptors may also have overlapping yet distinct functional characteristics. In analogy, retinoic acid receptors (RARs) and retinoid X receptors (RXRs) have similar amino acid sequence identities in the DBD and LBD region as the VDR and VDRRγ. RARs and RXRs have been shown to have distinct functional similarities such that both receptors bind 9-cis retinoic acid and have overlapping DNA-binding specificities and accordingly regulate overlapping gene networks. Based on these findings, VDRRγ may be regulated by small chemical molecules similar in structure to known ligands for nuclear receptors but not necessarily identical to ligands for the 1α,25-dihydroxy vitamin D3 receptor. Furthermore, VDRRγ may regulate vitamin D3 responsive gene networks by binding to a Vitamin D responsive element (VDRE)-like DNA sequence. In the present application, the 1α,25-dihydroxy vitamin D3 receptor is abbreviated as the Vitamin D receptor (VDR).
- In the present invention, the substance affecting VDRR signal transduction can be any small chemical molecule of natural or synthetic origin, e.g. a carbohydrate such as an aromatic compound. The small molecule may have a molecular weight in the range of from about 100 up to about 500 Da. Suitably, the small chemical molecule has a molecular weight in the range of from 200 up to 400 Da. Preferably, the small chemical molecule has a molecular weight of about 300 Da.
- The human VDRRγ polypeptides, including VDRRγ-1 and VDRRγ-2, have been shown to be activated e.g. by pregnenolones and estradiol (weakly), but not by certain other steroid hormones such as cortisol, aldosterone, progesterone and estrogen, and most likely not by progestines and glucocorticoids. Thus, human VDRRγ is not activated by pregnenolone 16α-carbonitrile (PCN), a glucocorticoid antagonist. For this reason, human VDRRγ can also be designated human pregnenolone activated (nuclear) receptors (hPAR). Information about pregnenolone can be found e.g. in the Merck Index, 11th ed., Merck & Co., Inc. Rahway, N.J., USA, p. 7735, 1989.
- Activators for human VDRRγ polypeptides, including VDRRγ-1 and VDRRγ-2, (hPAR-1 and hPAR-2, respectively), include but are not limited to pregnenolones, such as pregnane-ones, pregnane-diones, pregnane-triones, and pregnane-diols, and androstanes, such as androstane-ols, and androstane-diols. Suitably, the pregnenolones are non-planar, particularly 5β-pregnanes.
- Specific examples of activators and possibly ligands for human VDRRγ polypeptides, including VDRRγ-1 and VDRRγ-2, are the following compounds, which are marketed by Sigma-Aldrich of Sweden:
- i) 5β-pregnane-3,20-dione
- ii) 3α-hydroxy-5β-pregnane-11,20-dione methanesulphonate
- iii) 5β-pregnane-3α,20β-diol
- iv) pregnenolone
- v) Pregn-4-eno[16,17-δ][2]isoxazolline-3,20-dione, 6α-methyl-3′-phenyl-, ethyl ether solvate
- vi) Pregna-1,4,9(11)-triene-3,20-dione, 21-[4-[6-methoxy-2-(4-morpholinyl)-4-pyrimidinyl]-1-piperazinyl]-16-methyl-, (16α)-
- vii) Estran-3-ol, 17-[[[3-(trifluoromethyl)phenyl]methyl]amino]-, (E)-2-butenedioate (1:1) (salt)
- viii) 9α-Fluoro-5α-androstane-11β,17β-diol
- ix) Spiro[5α-androstane-3,2′-benzothiazolin]-11-one, 17β-hydroxy-17-methyl-
- x) Spiro[pregnane-3,2′-thiazolidine]-4′-carboxylic acid, 11α-hydroxy-20-oxo-, sodium salt
- xi) 17β-Dimethylamino-17-ethynyl-5α-androstane-11β-ol
- xii) 6β-Hydroxy-3,5-cyclo-5α-pregnan-20-one, nitrite
- xiii) 3α-Hydroxy-5β-pregnane-11,20-dione, acetate, 20-O-(methylsulfonyl)-oxime
- xiv) 17α-Methyl-5α-androstane-11β,17-diol
- xv) 5β-Pregnane-3,11,20-trione, trioxime
- xvi) 3α-Hydroxy-5β-pregnane-11,20-dione, 20-hydazone with hydrazide of 1-(carboxymethyl) pyridinium chloride.
A possible use of a VDRRg antagonist, could be a synergistic co-administration of the VDRRg antagonist together with other drugs such as, but not limited to, HIV protease inhibitors and cyclosporin to inhibit the expression of CYP3A4 and thus increase the bioavailability of drugs with poor pharmacokinetics due to CYP3A4 metabolism. Genes coding for polypeptides, such as human vitamin D receptor related gamma (hVDRRg), may be cloned by incorporating a DNA fragment coding for the polypeptide into a recombinant DNA vehicle, e.g. a vector and transforming suitable prokaryotic or eukaryo-tic host cells. Such recombinant DNA techniques are well known and e.g. described in Methods in Enzymology, Academic Press, San Diego, Calif., USA (1994), vols. 65 and 68 (1979), and vols. 100 and 101 (1983). - The host cells for use in the present invention can be prokaryotic or eukaryotic, preferably eukaryotic cells. Suitable eukaryotic host cells include but are not limited to cells from yeast e.g. Saccharomyces, insect cells and mammalian cells such as Chinese Hamster Ovary (CHO), Baby Hamster Kidney (BHK), COS and the like. Suitable prokaryotic host cells include but are not limited to cells from Enterobacteriacea, e.g. E. coli, Bacillus and Streptomyces.
- The following Examples are provided for purposes of illustration only and are not to be construed as in any way limiting the scope of the present invention, which is defined by the appended claims.
- Expressed Sequence Tag (EST) databases were screened for nuclear receptor related sequences with a DNA-binding domain (DBD) profile of nuclear receptors. This search profile was created by multiple alignment of a selected set of nuclear receptor sub-domains followed by a statistical calculation to obtain a so called Hidden Markov Model (HMM) of different subfamily members of the nuclear receptor supergene family. The cDNA of one of the nuclear receptor related EST sequences identified (Incyte clone no 2211526) was analyzed in detail by sequencing. After DNA sequencing of the entire Incyte cDNA done (approximately 2200 basepairs) the clone was found to encode a putative ligand-binding domain (LBD) with 54% and 44% similarity to xONR-1 and to the vitamin D receptor (VDR), respectively. The cDNA of the Incyte clone was not full-length and did not encode a sequence corresponding to a complete DBD.
- 5-RACE (rapid amplification of cDNA ends) of random primed cDNA from human liver RNA (InVitrogen) followed by cloning and DNA sequencing showed that the 5′-part of the cDNA corresponding to the Incyte clone encoded a DBD characteristic for nuclear receptors and with 71% and 65% sequence similarity to xONR-1 and VDR, respectively. Multiple alignments in combination with evolutionary neighbor-joining tree analysis placed the polypeptide encoded by the cDNA (specified in
FIG. 1 ) in the group of VDRs (FIGS. 2 and 3 ) and was named human vitamin D receptor related gamma (VDRRg). The deduced amino acid sequence of VDRRg is given inFIG. 4 . - Multiple tissue northern blots (Clontech) was used to the expression pattern of VDRRg in adult human tissues. As shown in
FIG. 5 , VDRRg is abundantly expressed in small intestine, mucosal lining of colon and liver but not in several other tissues including spleen, thymus, prostate, testis, ovary, peripheral blood leukocytes, heart, brain, placenta, lung, skeletal muscle, kidney and pancreas. To investigate if VDRRγ was expressed at lower levels in any of the other tissues examined, the filter was exposed for an extended time (one week as compared to overnight). Even after this prolonged exposure (data not shown), expression could still only be detected in the same tissues and not in any of the other tissues examined. The restricted expression pattern of VDRRg suggest that this receptor is likely to have an important regulatory function in liver and intestine. - Transient transfections were performed to analyze if vitamin D3 activate the VDRRγ polypeptide. To this end, transient co-transfections of CV-1 cells were performed with expression plasmids encoding fusion proteins of the GAL4-DBD fused to the LBD of either the VDR or the VDRR together with a reporter-plasmid containing five GAL4 responsive elements upstream of the luciferase gene. After transfection, cells were treated with vehicle (DMSO) alone or with vitamin D3 for 48 hours followed by harvesting of the cells and measurement of the luciferase activity in cell extracts. As shown in
FIG. 6 , vitamin D3 (1 μM) transactivate the GAL4-DBD/VDR-LBD but not the corresponding GAL4-DBD/-VDRRγ-LBD polypeptide under these conditions. This indicates that the two receptors may have distinct ligand-biding specificities. - 5′-RACE (see Example 1) of cDNA from human liver RNA followed by cloning and DNA sequencing identified an additional human VDRRγ cDNA with alternatively spliced 5′-end (see
FIG. 7 ). The VDRRγ cDNAs are thus able to encode at least one alternative N-terminal variant (FIG. 8 ) in addition to the VDRRγ polypeptide shown inFIG. 4 . The polypeptides disclosed inFIG. 4 andFIG. 8 which correspond to the differentially spliced VDRRγ cDNAs are designated as VDRRγ-1 and VDRRγ-2, respectively. - Expression plasmids containing VDRRγ or RXRβ cDNAs were described using T7 polymerase and translated in vitro in TNT reticulocyte lysates (Promega, Madison, Wis., USA). To investigate the DNA-binding specificity of VDRRγ a native gel mobility assay was employed essentially as described (Berkenstam et al., Cell, 69, 401-412, 1992) in which in vitro translated VDRRγ was incubated in the presence or absence of in vitro translated RXRβ with different 32P-labelled direct repeats (DR-1 to DR-5) as indicated in
FIG. 9 . The direct repeats were derived from the DR-5 element in the RAR-β2 promoter (de Thé et al., Nature, 343, 177-180, 1990) and modified to be separated by one to five nucleotides (Pettersson et al., Mechanisms of Dev., 54, 1-13, 1995). Protein-DNA complexes were separated on native 5% polyacryl-amide/0.25×TBE gels followed by autoradiography. As shown inFIG. 9 , of the five DRs tested efficient VDRRγ binding could only be detected with DRs separated by three or four nucleotides and only in the presence of RXR. However, weaker RXR-dependent binding could also be observed to DR-2 and DR-1 elements. These results demonstrate that VDRRγ require RXR heterodimerisation for efficient DNA-binding to a specific subset of DRs. These results, however, do not exclude the possibility that VDRRγ may bind as a monomer, dimer or heterodimer to distinct but related DNA-sequences. Importantly, our results demonstrate that VDRRγ and other nuclear receptors including the VDR (e.g. Markose, E. R. et al., Proc. Natl. Acad. Sci. USA, 87, 1701-1705, 1990), THRs (e.g. Gronemeyer, H. and Moras, D., Nature, 375, 190-191, 1995), LXRs (e.g. Willy, P. J. et al., Genes. Dev., 9, 1033-1045, 1995), have distinct but overlapping DNA-sequence and thus may regulate overlapping gene networks. - Interestingly, the most closely related nuclear receptor called ONR-1 (in Smith et al., 1994, Nucleic Acids Res., 22, pp 66-71) or XOR6 (in WO 96/122390) have been reported to “bind well to a retinoic acid response element, bRARE” (p. 11, line 30 in WO 96/22390). However, although the novel nuclear receptor VDRRg reported herein has 71% amino acid similarity in the DBD as compared to XOR-6 (
FIG. 12 ), VDRRg does not appear to bind to the same bRARE sequence (DR-5 inFIG. 9 ). - For identifying activators or ligands for VDRRγ, a library of substances structurally biased towards different classes of activators and ligands for nuclear receptors were tested. The activation of VDRRγ was analyzed in a reporter gene assay in transiently Caco-2 (TC7) cells (Carriere et al, 1994). In this initial screen, the synthetic substances with ability to activate VDRRγ were found to be structurally similar to pregnenolones (data not shown). Based on these results, naturally occurring pregnenolone derivatives were examined for activation of VDRRγ. The results are shown in
FIG. 10 . As is evident fromFIG. 10 , VDRRγ was activated about 5 to 12 fold by pregnenolone, 5β-pregnane-3,20-dione, 5β-pregnane-3α,20β-diol and 3α-hydroxy-5β-pregnane-11,20-dione methanesulphonate. In contrast to the efficient activation observed by the 5β-pregnane-3,20-dione, the corresponding planar steroid derivative 5α-pregnane-3,20-dione did not activate the receptor. Other 5β-pregnanes also activated VDRRγ efficiently as opposed to all planar pregnenolone derivatives tested, as is also evident fromFIG. 10 . - Further experiments were performed to find out if pregnenolone 16α-carbonitrile (PCN), a glucocorticoid antagonist or dexamethasone are activators of VDRRγ. To this effect, Caco-2 cells were transfected as before with VDRRγ and the activation was analyzed after treatment of the cells with 10 μM PCN or dexamethasone. The results are shown in
FIG. 11 . As is evident fromFIG. 11 , VDRRγ was not activated by these substances, indicating that VDRRγ is not the human PCN receptor. This suggestion is corroborated by the observation that also the antiprogestin RU486 only caused a slight increase (two fold) in VDRRγ mediated reporter gene activity as is evident fromFIG. 11 . - Activators of XOR-6 (FIG. 3 in WO 96/22390) such as butyl 4-NH2 Benzoate did not activate VDRRg (data not shown) in similar reporter assays as used in WO 96/22390.
Claims (31)
1. A mammalian, preferably human, isolated or recombinant nucleic acid comprising a contiguous nucleic acid sequence encoding a vitamin D receptor related (VDRR) polypeptide.
2. An isolated or recombinant DNA/nucleic acid according to FIG. 1 or FIG. 7 or alleles thereof encoding a new VDRR polypeptide.
3. The nucleic acid according to claim 1 or claim 2 encoding the VDRR polypeptide containing a DNA-binding domain (DBD) comprising about 77 amino acids with 9 cysteine residues, wherein said DBD is characterized by the following amino acid sequence similarity:
(i) at least 60% amino acid sequence similarity with the DBD of hVDR; and
(ii) at least 65% amino acid sequence similarity with the DBD of xONR1.
4. The nucleic acid according to claim 3 , wherein said DBD is characterized by the following amino acid sequence similarity:
(i) about 65% amino acid sequence similarity with the DBD of hVDR; and
(ii) about 71% amino acid sequence similarity with the DBD of xONR1.
5. The nucleic acid according to any previous claim, encoding the VDRR polypeptide, wherein the ligand-binding domain (LBD) of said polypeptide is characterized by the following amino acid sequence similarity, relative to the LBDs of hVDR and xONR1, respectively:
(i) at least about 30% amino acid sequence similarity with the LBD of hVDR; and
(ii) at least about 40% amino acid sequence similarity with the LBD of xONR1.
6. The nucleic acid according to claim 5 , wherein said LBD is characterized by the following amino acid sequence similarity:
(i) at least 35% amino acid sequence similarity with the LBD of hVDR; and
(ii) at least 45% amino acid sequence similarity with the LBD of xONR1.
7. The nucleic acid according to claim 6 , wherein said LBD is characterized by the following amino acid sequence similarity:
(i) about 42% amino acid sequence similarity with the LBD of hVDR; and
(ii) about 54% amino acid sequence similarity with the LBD of xONR1.
8. The nucleic acid according to any previous claim, wherein said nucleic acid sequence is that given in FIG. 1 or FIG. 7 or alleles thereof.
9. The nucleic acid according to claim 8 , wherein said nucleic acid sequence is the same or substantially the same as given in FIG. 1 or FIG. 7 .
10. A nucleic acid probe for the detection of a nucleic acid sequence encoding a VDRR polypeptide in a sample.
11. The nucleic acid probe according to claim 10 , wherein said probe comprises at least 14 contiguous nucleotides of the nucleic acid sequence given in FIG. 1 or FIG. 7 .
12. A method for identifying clones encoding a VDRR polypeptide said method comprising screening a genomic or cDNA library with a nucleic acid probe according to claim 10 or 11 under low stringency hybridization conditions, and identifying those clones which display a substantial degree of hybridization to said probe.
13. An expression vector comprising a nucleic acid according to any of claims 1-9.
14. A cell containing a nucleic acid according to any of claims 1-9.
15. A cell containing an expression vector according to claim 14 .
16. A process for recombinant production of a VDRR polypeptide, said process comprising expressing the nucleic acid of any of claims 1 to 9 in a suitable host cell.
17. The process according to claim 16 , wherein the host cell is eukaryotic.
18. An isolated or recombinant mammalian, preferably human, VDRR polypeptide.
19. The isolated or recombinant VDRR polypeptide according to claim 18 comprising the amino acid sequence substantially the same or the same as given in FIG. 4 or FIG. 8 .
20. A method to produce specific monoclonal and polyclonal antibodies to the polypeptide according to any of claims 18 and 19 comprising the injection of the protein to a mammalian.
21. A pharmaceutical formulation comprising an isolated or recombinant VDRR polypeptide according to any of claim 18 and 19 , and one or more therapeutically acceptable excipients.
22. A method for identifying a ligand to a VDRR according to any of claim 18 and 19 , by a cell-based reporter assay, transgenic-animal reporter assay or in vitro-binding assay.
23. A method for identifying a substance for treatment of a condition affected by a VDRR polypeptide according to any of claim 18 and 19 , comprising screening for an agonist or an antagonist of VDRR polypeptide signal transduction to be used for treating metabolic, proliferative or inflammatory conditions.
24. A mammalian, preferably human, VDRR polypeptide according to any of claim 18 and 19 for use as a medicament.
25. Use of a substance affecting VDRR, according to any of claim 18 and 19 , signal transduction, such as an agonist or an antagonist of VDRR polypeptide signal transduction, for the manufacture of a medicament for treating metabolic, proliferative or inflammatory conditions.
26. Use of a substance affecting VDRR, according to any of claim 18 and 19 , signal transduction for the manufacture of a medicament for treating obesity, diabetes, anorexia, lipoprotein defects, hyperlipidemia, hypercholesteremia or hyperlipoproteinemia.
27. Use of a substance affecting VDRR, according to any of claim 18 and 19 , signal transduction for the manufacture of a medicament for treating osteoporosis, rheumatoid arthritis, benign and malign tumors, hyperproliferative skin disorders or hyperparathyroidism.
28. Use according to any of claims 25-27, wherein the substance affecting VDRR signal transduction is a chemical molecule of natural or synthetic origin with a molecular weight in the range of from about 100 up to about 500 Da, preferably with a molecular weight of about 300 Da.
29. A method for treating metabolic, proliferative or inflammatory conditions comprising introducing into a mammal a nucleic acid vector according to claim 13 encoding for expression of a VDRR polypeptide and wherein said nucleic acid vector is capable of transforming a cell in vivo and expressing said polypeptide in said transformed cell.
30. A method for treatment of a metabolic, proliferative or inflammatory condition by administration of a therapeutically effective amount of a substance affecting VDRR, according to any of claim 18 and 19 , signal transduction.
31. The method according to claim 30 , wherein the substance affecting VDRR signal transduction is a chemical molecule of natural or synthetic origin with a molecular weight in the range of from about 100 up to about 500 Da, preferably with a molecular weight of about 300 Da.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11/369,263 US20060228778A1 (en) | 1997-10-14 | 2006-03-06 | Novel vitamin D receptor related polypeptides, nucleic acid sequence encoding the same and uses thereof |
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
SE9703745-1 | 1997-10-14 | ||
SE9703745A SE9703745D0 (en) | 1997-10-14 | 1997-10-14 | New receptors |
US6737397P | 1997-12-03 | 1997-12-03 | |
SE9801148A SE9801148D0 (en) | 1997-10-14 | 1998-03-31 | New receptors |
SE9801148-9 | 1998-03-31 | ||
US09/143,828 US7118885B2 (en) | 1997-10-14 | 1998-08-31 | Nucleic acid encoding vitamin D receptor related polypeptide |
US11/369,263 US20060228778A1 (en) | 1997-10-14 | 2006-03-06 | Novel vitamin D receptor related polypeptides, nucleic acid sequence encoding the same and uses thereof |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/143,828 Division US7118885B2 (en) | 1997-10-14 | 1998-08-31 | Nucleic acid encoding vitamin D receptor related polypeptide |
Publications (1)
Publication Number | Publication Date |
---|---|
US20060228778A1 true US20060228778A1 (en) | 2006-10-12 |
Family
ID=27484800
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/143,828 Expired - Fee Related US7118885B2 (en) | 1997-10-14 | 1998-08-31 | Nucleic acid encoding vitamin D receptor related polypeptide |
US11/369,263 Abandoned US20060228778A1 (en) | 1997-10-14 | 2006-03-06 | Novel vitamin D receptor related polypeptides, nucleic acid sequence encoding the same and uses thereof |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/143,828 Expired - Fee Related US7118885B2 (en) | 1997-10-14 | 1998-08-31 | Nucleic acid encoding vitamin D receptor related polypeptide |
Country Status (1)
Country | Link |
---|---|
US (2) | US7118885B2 (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5508164A (en) * | 1990-10-29 | 1996-04-16 | Dekalb Genetics Corporation | Isolation of biological materials using magnetic particles |
US6391847B1 (en) * | 1995-01-17 | 2002-05-21 | The Salk Institute For Biological Studies | Method, polypeptides, nucleotide sequence of XOR-6, a vitamin D-like receptor from xenopus |
US6756491B2 (en) * | 1998-01-09 | 2004-06-29 | The Salk Institute For Biological Studies | Steroid-activated nuclear receptors and uses therefor |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0407462A4 (en) | 1988-03-30 | 1992-02-05 | Arch Development Corporation | Dna binding proteins including androgen receptor |
AU2579692A (en) | 1991-09-17 | 1993-04-27 | Salk Institute For Biological Studies, The | Receptor of the thyroid/steroid hormone receptor superfamily |
US5756448A (en) | 1992-02-26 | 1998-05-26 | The General Hospital Corporation | Constitute activator of retinoid (CAR) receptor polypeptides |
AU5426596A (en) | 1995-05-16 | 1996-11-29 | Salk Institute For Biological Studies, The | Modulators for new members of the steroid/thyroid superfamily of receptors |
-
1998
- 1998-08-31 US US09/143,828 patent/US7118885B2/en not_active Expired - Fee Related
-
2006
- 2006-03-06 US US11/369,263 patent/US20060228778A1/en not_active Abandoned
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5508164A (en) * | 1990-10-29 | 1996-04-16 | Dekalb Genetics Corporation | Isolation of biological materials using magnetic particles |
US6391847B1 (en) * | 1995-01-17 | 2002-05-21 | The Salk Institute For Biological Studies | Method, polypeptides, nucleotide sequence of XOR-6, a vitamin D-like receptor from xenopus |
US6756491B2 (en) * | 1998-01-09 | 2004-06-29 | The Salk Institute For Biological Studies | Steroid-activated nuclear receptors and uses therefor |
US6809178B2 (en) * | 1998-01-09 | 2004-10-26 | The Salk Institute For Biological Studies | Steroid-activated nuclear receptors and uses therefor |
Also Published As
Publication number | Publication date |
---|---|
US7118885B2 (en) | 2006-10-10 |
US20030032790A1 (en) | 2003-02-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Ito et al. | DAX-1 inhibits SF-1-mediated transactivation via a carboxy-terminal domain that is deleted in adrenal hypoplasia congenita | |
Oberkofler et al. | Potentiation of liver X receptor transcriptional activity by peroxisome-proliferator-activated receptor gamma co-activator 1alpha | |
Harding et al. | The orphan receptor Rev-ErbAα activates transcription via a novel response element | |
Zhang et al. | The mechanism of action of thyroid hormones | |
Dussault et al. | A structural model of the constitutive androstane receptor defines novel interactions that mediate ligand-independent activity | |
Christian et al. | Functional association of PR and CCAAT/Enhancer-Binding Proteinβ isoforms: Promoter-dependent cooperation between PR-B and Liver-Enriched Inhibitory Protein, or Liver-Enriched Activatory Protein and PR-A in human endometrial stromal cells | |
US6387673B1 (en) | Compounds useful for the modulation of processes mediated by nuclear hormone receptors, methods for the identification and use of such compounds | |
Wyszomierski et al. | Cooperative effects of STAT5 (signal transducer and activator of transcription 5) and C/EBP β (CCAAT/enhancer-binding protein-β) onβ-casein gene transcription are mediated by the glucocorticoid receptor | |
Blumberg et al. | BXR, an embryonic orphan nuclear receptor activated by a novel class of endogenous benzoate metabolites | |
EP1162264A2 (en) | Chimeric hormone receptor | |
Murphy et al. | Differential regulation of transcription by the NURR1/NUR77 subfamily of nuclear transcription factors | |
AU732079B2 (en) | Novel vitamin D receptor related polypeptides, nucleic acid sequence encoding the same and uses thereof | |
JP2006068010A (en) | Retinoid x receptor-interacting polypeptides and related molecule and method | |
Delort et al. | TAXI/UAS: A molecular switch to control expression of genes in vivo | |
Gessi et al. | Glucocorticoids' pharmacology: past, present and future | |
Souza et al. | Helix 12 dynamics and thyroid hormone receptor activity: experimental and molecular dynamics studies of Ile280 mutants | |
Le Douarin et al. | Ligand-dependent interaction of nuclear receptors with potential transcriptional intermediary factors (mediators) | |
CN110507655B (en) | Application of compound FG-4592 in preparation of pharmaceutical preparation for treating thyroid hormone receptor mediated diseases | |
Kardassis et al. | Transactivation of the Human Apolipoprotein CII Promoter by Orphan and Ligand-dependent Nuclear Receptors: The Regulatory Element CIIC is a Thyroid Hormone Response Element | |
US7118885B2 (en) | Nucleic acid encoding vitamin D receptor related polypeptide | |
JP2003520022A (en) | DNA encoding human vanilloid receptor VR1 | |
Yasumo et al. | Nuclear receptor binding factor-2 (NRBF-2), a possible gene activator protein interacting with nuclear hormone receptors | |
US6277976B1 (en) | Or-1, an orphan receptor belonging to the nuclear receptor family | |
Leister et al. | Apoptosis antagonizing transcription factor AATF is a novel coactivator of nuclear hormone receptors | |
Nakajima et al. | Characterization of the activation function-2 domain of the human 1, 25-dihydroxyvitamin D3 receptor |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |