Claims
- 1. A nucleic acid comprising a nucleotide sequence encoding a mammalian biogenic amine receptor.
- 2. A nucleic acid according to claim 1 wherein the mammalian biogenic amine receptor is a human trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No. 2.
- 3. A nucleic acid according to claim 1 wherein the mammalian biogenic amine receptor is a rat trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 4.
- 4. A homogeneous composition of a mammalian trace amine receptor or derivative thereof having a molecular weight of about 39 kilodaltons and an amino acid sequence identified by (SEQ ID No.: 2).
- 5. A homogeneous composition of a mammalian trace amine receptor or derivative thereof having a molecular weight of about 38 kilodaltons and an amino acid sequence identified by (SEQ ID No.: 4).
- 6. A nucleic acid hybridization probe comprising a nucleotide sequence identified by Seq ID No. 1.
- 7. A nucleic acid hybridization probe comprising a nucleotide sequence identified by Seq ID No. 3.
- 8. A recombinant expression construct comprising a nucleic acid having a nucleotide sequence encoding a mammalian trace amine receptor according to claim 1, wherein the construct is capable of expressing the receptor in a transformed culture of eukaryotic or prokaryotic cells.
- 9. A recombinant expression construct according to claim 8 wherein the mammalian biogenic amine receptor is a human trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 2.
- 10. A recombinant expression construct according to claim 8 wherein the mammalian biogenic amine receptor is a rat trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No. 4.
- 11. A cell culture transformed with the recombinant expression construct of claim 8, wherein the transformed cell culture expresses the mammalian trace amine receptor.
- 12. A cell culture transformed with the recombinant expression construct of claim 9, wherein the transformed cell culture expresses the human trace amine receptor.
- 13. A cell culture transformed with the recombinant expression construct of claim 10, wherein the transformed cell culture expresses the rat trace amine receptor.
- 14. A method of screening a compound for binding to a mammalian trace amine receptor in cells expressing the receptor, the method comprising the steps of:
(a) transforming a host cell with a recombinant expression construct encoding a mammalian trace amine receptor according to claim 1, wherein the cells of the transformed cell culture express the receptor, and (b) assaying the transformed cell culture with the compound to determine whether the compound binds to the mammalian trace amine receptor.
- 15. The method of claim 14 wherein the mammalian trace amine receptor is a human trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 2.
- 16. The method of claim 14 wherein the mammalian trace amine receptor is a rat trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 4.
- 17. A method of claim 14 comprising the additional step of:
(c) comparing binding of the compound with binding of additional compounds that are known to bind to mammalian trace amine receptors, wherein said additional compounds comprise naturally-occurring and synthetic receptor agonists and antagonists.
- 18. A method of screening a compound for competitive binding to a mammalian trace amine receptor in cells expressing the receptor, the method, comprising the following steps:
(a) transforming a host cell with a recombinant expression construct encoding a mammalian trace amine receptor according to claim 1, wherein the cells of the transformed cell culture express the receptor, (b) assaying the transformed cell with the compound in the presence and in the absence of an agonist for the receptor; and (c) determining whether the compound competes with the agonist for binding to the receptor.
- 19. The method of claim 18 wherein the mammalian trace amine receptor is a human trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 2.
- 20. The method of claim 18 wherein the mammalian trace amine receptor is a rat trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 4.
- 21. The method of claim 18, wherein the compound is detectably-labeled.
- 22. The method of claim 18, wherein the receptor agonist is detectably-labeled.
- 23. The method of claim 18, wherein the mammalian trace amine receptor competitor is quantitatively characterized by assaying the transformed cell culture with varying amounts of the competitor in the presence of a detectably-labeled receptor agonist and measuring the extent of competition with receptor binding thereby.
- 24. A method of screening a compound to determine if the compound is an inhibitor of a mammalian trace amine receptor in cells expressing the receptor, the method comprising the following steps:
(a) transforming a host cell with a recombinant expression construct encoding a mammalian trace amine receptor according to claim 1, wherein the cells of the transformed cell culture express the receptor; (b) assaying the transformed cell culture with the compound to determine whether the compound is capable of inhibiting trace amine receptor binding by a receptor agonist.
- 25. The method of claim 24 wherein the mammalian trace amine receptor is a human trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.: 2.
- 26. The method of claim 24 wherein the mammalian trace amine receptor is a rat trace amine receptor and the nucleotide sequence of the nucleic acid encodes an amino acid sequence identified by SEQ ID No.:4.
- 27. The method of claim 24, wherein the compound is detectably-labeled.
- 28. The method of claim 24, wherein the receptor agonist is detectably-labeled.
- 29. The method of claim 24, wherein the trace amine receptor inhibitor is quantitatively characterized by assaying the transformed cell culture with varying amounts of the inhibitor in the presence of a detectably-labeled trace amine receptor agonist and measuring the et of inhibition of agonist binding thereby.
- 30. A nucleic acid comprising a nucleotide sequence encoding a mammalian trace amine receptor that hybridizes to a nucleic acid having a nucleotide sequence identified by Seq. ID No. 1, under conditions of 37° C. in a buffer comprising 50% formamide, 1% sodium dodecyl sulfate, 5×SSC, 50 μg/mL denatured salmon sperm DNA, and 5× P-buffer comprising 0.25M Tris, pH 7.5, 0.5% sodium pyrophosphate, 0.5%. SDS, 1% bovine serum albumin, 1% polyvinylpyrrolidone and 1% Ficoll.
- 31. A nucleic acid according to claim 30, wherein the nucleic acid hybridizes to a nucleic acid having a nucleotide sequence identified by Seq. ID No. 1, under washing conditions of 10 minutes at room temperature in a wash solution of 2×SSC/1% SDS, followed by 10 min at 60° C. in 2×SSC/1% SDS, followed by 5 min at 60° C. in 0.5×SSC/1% SDS, followed by 5 min at 60° C. in 0.5×SC/1% SDS.
- 32. A nucleic acid comprising a nucleotide sequence encoding a mammalian trace amine receptor that hybrids to a nucleic acid having a nucleotide sequence identified by Seq. ID No. 3, under conditions of 37° C in a buffer comprising 50% formamide, 1% sodium dodecyl sulfate, 5×SSC, 50 μg/mL denatured salmon sperm DNA, and 5× P-buffer comprising 0.25M Tris, pH 7.5, 0.5% sodium pyroposphate, 0.5% SDS, 1% bovine serum albumin, 1% polyvinylpyrrolidone and 1% Ficoll.
- 33. A nucleic acid according to claim 32, wherein the nucleic acid hybridizes to a nucleic acid having a nucleotide sequence identified by Seq. ID No. 3, under washing conditions of 10 minutes at room temperature in a wash solution of 2×SSC/1% SDS, followed by 10 min at 60° C. in 2×SSC/1% SDS, followed by 5 min at 60° C. in 0.5×SC/1% SDS.
- 34. A cell membrane preparation comprising a mammalian trace amine receptor or derivative thereof having a molecular weight of about 39 kilodaltons and an amino acid sequence identified by SEQ ID No.: 2.
- 35. A cell membrane preparation comprising a mammalian trace amine receptor or derivative thereof having a: molecular weight of about 38 kilodaltons and an amino acid sequence identified by SEQ ID No.: 4.
- 36. A cytosolic preparation comprising a mammalian trace amine receptor or derivative thereof having a molecular weight of about 39 kilodaltons and an ammo acid sequence identified by SEQ ID No.: 2.
- 37. A cytosolic preparation comprising a mammalian trace amine receptor or derivative thereof having a molecular weight of about 38 kilodaltons and an amino acid sequence identified by SEQ ID No.: 4.
- 38. The method of claim 17, 18, or 24 wherein the agonist is a neurotransmitter.
- 39. The method of claim 38 wherein the neurotransmitter is p-tyramine, phenylethylamine, tryptamine, octopamine, synephrine, dopamine, serotonin, or m-tyramine.
- 40. The method of claim 17, 18, or 24 wherein the agonist is a drug.
- 41. The method of claim 40 wherein the drug is O-phenethylamine (EA), hordenine, L-tyrosinol, SR-amphetamine (+and −), 4-OH—R(−)-amphetamine, methamphetamine (+and −), (±)DOI, phenelzine, tranylcypromine, 3-methoxytyramine>dopamine, 3,4-DiMeO-PEA>Mescaline, (±)MDMA, 3,4-dihydroxybenzylguanidine, 3-phenylpropylamine, 1-methyl-3-phenylpropylamine, N,N-dimethylpropiophenone, N-phenylethylenediamine, kynuramine, 4-phenylbutylamine, tryptamine, 2-thiopheneethylamine, betahistine, 2>4>3-pyridylethylamine, 1-phenylpiperazine, 1-napthyl)piperazine, 1,2,3,4-tetrahydroisoquinoline, (i)saisolinol hydrocotamine, nomifensine, R(−)apomorphine, S(+)2-aminotetralin, R(−)2-aminotetralin, (±)2-amino-1,2-dihydronapthalene, (±)3A6HC, (±)2-aminoindan, MPTP, 4phenyl-1,2,3,6-tetrahydropyridine, tolazoline, naphazoline, phentolamine, agroclavine, bromocriptine, lisuride, d-LSD, metergoline, (±)fenfluramine, fenspiride, 2-phenyl-2-imidazoline, methylphenidate, pargyline, 2,2-diphenylethylamine, trans-cinnamyl-piperazine, 1-benzyl-piperidine, rimantidine, tripelennamine, tryptamine/5-MeO-D MT, forskolin, amphetamine/phentermine, cyproheptadine, dopamine, dihydroergotamine, fenoterol, HVA:D1 receptor, imidazoline/naphazoline, imidazoline/oxymetazoline, imidazoline/phentolamine, imidazoline/tolazoline, isoproterenol, metanephrine DL, methamphetamine/2-MeO, octopamine, PEA/2-amino 4-OH, PEA/3,4 dimethoxy, 4-OH-PEA/3-MeO, 4-amino-PEA, 4-methoxy-PEA, AEBSF-PEA, phenylephrine, piperazine/mCPP, piperazine/TFMPP, phenylpiperazine, prenylamine, 2-(2aminoethyl)pyradine, 3-(2aminoethyl)pyradine, 4-(2aminoethyl)pyradine, ritodrine, synephrine, tetralin/ADIN/6,7,5-Fluoro-tryptamine, N,N-dimethyl-tryptamine, tryptophanol (±), m-tyramine, p-tyramine, or 3-hydroxtyramine.
- 42. A pharmaceutical composition comprising the compound of claim 17, 18, or 24 in admixture with a pharmaceutically acceptable carrier.
- 43. A method of reducing sympathomimetic effects of enhanced trace amine dependent synaptic transmission in a mammal comprising administering to said mammal a therapeutically effective amount of the pharmaceutical composition of claim 42.
- 44. The method of claim 43 wherein the mammal is a human.
- 45. A method of treating peripheral effects of a drug that binds to or affects the binding to trace amine receptors in a mammal comprising administering to said mammal a therapeutically effective amount of the pharmaceutical composition of claim 42.
- 46. The method of claim 45 wherein the peripheral effect is hyperthermia, rapid heart rate, high blood pressure, migraine, cardiac arrythmia, seizures, coma and diabetes.
- 47. The method of claim 45 wherein the mammal is a human.
- 48. A method of treating a pathological condition associated with elevated levels of trace amines in a mammal comprising administering to said mammal a therapeutically effective amount of the pharmaceutical composition of claim 42.
- 49. The method of claim 48 wherein the pathological condition is schizophrenia.
- 50. The method of claim 48 wherein the pathological condition is depression.
- 51. The method of claim 48 wherein the pathological condition is shock, hypertension, cardiac arrhythmia, asthma, migraine headache, psychosis, anaphylactic reactions and iatrogenic conditions.
- 52. The method of claim 48 wherein the mammal is a human.
- 53. A method of treating drug addiction in a mammal comprising administering to said mammal a therapeutically effective amount of the pharmaceutical composition of claim 42.
- 54. The method of claim 53 wherein the drug is β-phenethylamine (PEA), hordenine, L-tyrosinol, S,R-amphetamine (+and ±), 4-OH—R(−)-amphetamine, methamphetamine (+and −), (±)DOI, phenelzine, tranylcypromine, 3-methoxytryramine>dopamine, 3,4-DMeO-PEA>Mescaline, (±)MDMA, 3,4-dihydroxybenzylguanidine, 3-phenylpropylamine, 1-methyl-3-phenylpropylamine, N,N-dimethylpropiophenone, N-phenylethylenediamine, kynuramine, 4-phenylbutylamine, tryptamine, 2-thiopheneethylamine, betahistine, 2>4>3-pyridylethylamine, 1-phenylpiperazine, 1-(1-napthyl)piperazne, 1,2,3,4-tetrahydroisoquinoline, (±)salsolinol, hydrocotamine, nomifensine, R(−)apomorphine, S(+)2-aminotetralin, R(−)2-aminotetralin, (±)2-amino-1,2 dihydronaptalene, (±)3A6HC, (±)2-aminoindan, MPTP, 4-phenyl-1,2,3,6-tetrahydropyridine, tolazoline, naphazoline, phentolamine, agroclavine, bromocriptine, lisuride, d-LSD, metergoline, (±)fenfluramine, fenspiride, 2-phenyl-2-imidazoline, methylphenidate, pargyline, 2,2-diphenylethylamine, trans-cinnamyl-piperazine, 1-benzyl-piperdine, rimantidine, tripelennamine, tryptamine/5-MeO-DMT, forskolin, amphetamine/phentermine, cyproheptadine, dopamine, dlhydroergotamine, fenoterol, HVA:D1 receptor, imidazoline/naphazoline, imidazoline/oxymetazoline, imidazoline/phentolamine, imidazoline/tolazoline, isoproterenol, metanephrine DL, methamphetamine/2-MeO, octopamine, PEA/2-amino 40E, PEA/3,4 dimethoxy, 4-OH-PEA/3-MeO, 4-amino-PEA, 4-methoxy-PEA, AEBSF-PEA, phenylephrine, piperazine/mCPP, piperazine/TFMPP, phenylpiperazine, prenylamine, 2-(2aminoethyl)pyradine, 3-(2aminoethyl)pyradine, 4-(2aminoethyl)pyradine, ritodrine, synephrine, tetralin/ADTN/6,7,5-Fluoro-tryptamine, N,N-dimethyl-tryptamine, tryptophanol (±), m-tyramine, p-tyramine, or 3-hydroxyyramine.
- 55. The method of claim 53 wherein the mammal is a human.
Government Interests
[0001] This invention was made with government support under National Institute of Health grants DA08562. The government has certain rights to this invention.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US01/28455 |
9/12/2001 |
WO |
|