Claims
- 1. A transgenic mouse expressing a human CAR receptor.
- 2. The transgenic mouse of claim 1, wherein said mouse does not express a murine CAR receptor having at least 60% of the naturally occurring murine CAR receptor activity.
- 3. The transgenic mouse of claim 1, wherein said mouse does not express a murine CAR receptor.
- 4. The transgenic mouse of claim 1, wherein said human CAR receptor induces the expression of a CAR target gene.
- 5. The transgenic mouse of claim 4, wherein said CAR target gene is CYP2B 10 or CYP2B6.
- 6. The transgenic mouse of claim 5, wherein said CAR target gene is CYP2B10.
- 7. The transgenic mouse of claim 4, wherein said CAR target gene is CYP3A11 or CYP3A4.
- 8. The transgenic mouse of claim 4, wherein said human CAR receptor is activated by phenobarbital.
- 9. The transgenic mouse of claim 4, wherein activity of said human CAR receptor is increased by less than 50% by 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene.
- 10. The transgenic mouse of claim 4, wherein phenobarbital increases an activity of said human CAR receptor by at least 2 fold more than the corresponding amount of 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene increases said activity of said human CAR receptor.
- 11. The transgenic mouse of claim 4, wherein said human CAR receptor is activated by acetaminophen.
- 12. The transgenic mouse of claim 1, wherein the mRNA of said human CAR receptor comprises a bovine growth hormone polyadenylation sequence.
- 13. The transgenic mouse of claim 1, wherein the nucleic acid encoding said human CAR receptor is operably linked to an intronic sequence of a rabbit β-globin nucleic acid.
- 14. The transgenic mouse of claim 13, wherein said intronic sequence increases the expression of said human CAR receptor.
- 15. A transgenic mouse expressing a human CAR receptor, wherein said transgenic mouse exhibits an induction of a xenobiotic response in the presence of a xenobiotic inducer.
- 16. The transgenic mouse of claim 15, wherein said xenobiotic response comprises an induction or activation of a CAR target gene.
- 17. The mouse of claim 16, wherein said CAR target gene is CYP2B10.
- 18. The transgenic mouse of claim 15, wherein said xenobiotic inducer is phenobarbitol.
- 19. The transgenic mouse of claim 15, wherein said transgenic mouse does not express murine CAR.
- 20. The transgenic mouse of claim 15, wherein said human CAR receptor is the human CAR ligand binding domain.
- 21. A transgenic mouse comprising a mutation in a CAR gene, wherein said comprises a reduction or absence of CAR receptor activity.
- 22. The transgenic mouse of claim 21, wherein said transgenic mouse exhibits a decrease or absence in induction, expression, or activity of a CAR target genes in response to a xenobiotic inducer.
- 23. The transgenic mouse of claim 21, wherein said xenobiotic inducer is phenobarbitol or 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene.
- 24. The transgenic mouse of claim 21, wherein said transgenic mouse does not exhibit hyperplasia or hypertrophy of the liver upon treatment with an xenobiotic inducer.
- 25. The transgenic mouse of claim 24, wherein said xenobiotic inducer is phenobarbitol or 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene.
- 26. The transgenic mouse of claim 21, wherein said transgenic mouse exhibits decreased metabolism of zoxazolamine.
- 27. The transgenic mouse of claim 21, wherein said transgenic mouse exhibits a resistance to acetaminophen toxicity.
- 28. A cell or tissue isolated from the transgenic mouse of claim 15 or claim 21.
- 29. A screening method for determining whether a compound activates a CAR receptor, said method comprising the steps of:
(a) administering a compound to a transgenic mouse expressing a human CAR receptor; and (b) measuring induction of a CAR target gene, whereby said compound is determined to activate said CAR receptor if said compound mediates induction of said CAR target gene.
- 30. The method of claim 29, wherein step (a) further comprises administering a CAR receptor inverse agonist to said mouse expressing said human CAR receptor.
- 31. The method of claim 30, wherein said CAR receptor inverse agonist is clotrimazole.
- 32. A screening method for determining whether a compound inhibits a CAR receptor, said method comprising the steps of:
(a) administering said compound to a transgenic mouse expressing a human CAR receptor; and (b) measuring expression of a CAR target gene in the presence and absence of said compound, whereby said compound is determined to inhibit said CAR receptor if said compound decreases said expression of said CAR target gene.
- 33. The method of claim 32, wherein step (a) further comprises administering a CAR receptor agonist to said mouse expressing said human CAR receptor.
- 34. The screening method of claim 33, wherein said CAR receptor agonist is administered after said compound.
- 35. A screening method for determining whether a compound modulates the activity of a CAR receptor, said method comprising the steps of:
(a) administering said compound to a transgenic mouse expressing a human CAR receptor; and (b) measuring a physiological effect mediated by the administration of said compound, whereby said compound is determined to modulate the activity of said CAR receptor if the magnitude of said physiological effect in said mouse expressing said human receptor differs from that in a mouse comprising a mutation that reduces CAR receptor activity.
- 36. The method of claim 35, wherein said measuring said physiological effect comprises measuring the toxicity or activity mediated by the administration of said compound or measuring the half-life of said compound.
- 37. The method of claim 36, wherein said toxicity or activity is mediated by a metabolite of said compound.
- 38. A screening method for determining whether the metabolism of a compound is regulated by modulation of the activity of a CAR receptor, said method comprising the steps of:
(a) administering said compound to a transgenic mouse expressing a human CAR receptor; and (b) measuring the rate of metabolism of said compound, whereby said metabolism of said compound is determined to be regulated by modulation of the activity of said CAR receptor if said rate of metabolism is faster in said mouse expressing said human receptor than in a mouse comprising a mutation that reduces CAR receptor activity.
- 39. The method of claim 38, wherein said measuring said rate of metabolism comprises measuring the toxicity or activity mediated by the administration of said compound, measuring the half-life of said compound, or measuring the serum level of a liver enzyme.
- 40. A screening method for determining whether the metabolism of a first compound is modulated by a second compound, said method comprising the steps of:
(a) administering said first compound in the presence and absence of said second compound to a transgenic mouse expressing a human CAR receptor; and (b) in the presence and absence of said second compound, measuring a physiological effect that is mediated by the administration of said first compound, whereby said second compound is determined to modulate the metabolism of said first compound if said second compound effects a change in said physiological effect mediated by said administration of said first compound.
- 41. The method of claim 40, wherein said measuring said physiological effect comprises measuring the toxicity or activity mediated by the administration of said first compound or measuring the half-life of said first compound.
- 42. The method of claim 41, wherein said toxicity or activity is mediated by a metabolite of said first compound.
- 43. The method of claim 41, wherein step (b) comprises measuring the half-life of said first compound in the presence and absence of said second compound, whereby said second compound is determined to activate the metabolism of said first compound if said second compound decreases said half-life, or said second compound is determined to inhibit the metabolism of said first compound if the said second compound increases said half-life.
- 44. The method of claim 29, 32, 35, 38, or 40, wherein said mouse expressing said human CAR receptor does not express a substantially active murine CAR receptor.
- 45. A screening method for determining whether a compound activates a CAR receptor, said method comprising the steps of:
(a) administering a compound to a mouse, said mouse comprising a mutation that reduces CAR receptor activity; and (b) measuring induction of a CAR target gene, whereby said compound is determined to activate said CAR receptor if said induction is smaller in said mouse comprising said mutation than in a mouse having wild-type CAR receptor activity.
- 46. The method of claim 45, wherein step (a) further comprises administering a CAR receptor inverse agonist to said mouse comprising said mutation.
- 47. The method of claim 46, wherein said CAR receptor inverse agonist is androstanol.
- 48. A screening method for determining whether a compound inhibits a CAR receptor, said method comprising the steps of:
(a) administering said compound to a mouse, said mouse comprising a mutation that reduces CAR receptor activity; and (b) measuring expression of a CAR target gene in the presence and absence of said compound, whereby said compound is determined to inhibit said CAR receptor if the decrease in said expression effected by said compound is smaller in said mouse comprising said mutation than in a mouse having wild-type CAR receptor activity.
- 49. The method of claim 48, wherein step (a) further comprises administering a CAR receptor agonist to said mouse comprising said mutation.
- 50. The method of claim 49, wherein said CAR receptor agonist is TCPOBOP, and said TCPOBOP is administered after said compound.
- 51. A screening method for determining whether a compound modulates the activity of a CAR receptor, said method comprising the steps of:
(a) administering said compound to a mouse, said mouse comprising a mutation that reduces CAR receptor activity; and (b) measuring a physiological effect mediated by the administration of said compound, whereby said compound is determined to modulate the activity of said CAR receptor if the magnitude of said physiological effect in said mouse comprising said mutation differs from that in a mouse having wild-type CAR receptor activity.
- 52. The method of claim 51, wherein said measuring said physiological effect comprises measuring the toxicity or activity mediated by the administration of said compound or measuring the half-life of said compound.
- 53. The method of claim 52, wherein said toxicity or activity is mediated by a metabolite of said compound.
- 54. A screening method for determining whether the metabolism of a compound is regulated by modulation of the activity of a CAR receptor, said method comprising the steps of:
(a) administering said compound to a mouse, said mouse comprising a mutation that reduces CAR receptor activity; and (b) measuring the rate of metabolism of said compound, whereby said metabolism of said compound is determined to be regulated by modulation of the activity of said CAR receptor if said rate of metabolism is slower in said mouse comprising said mutation than in a mouse having wild-type CAR receptor activity.
- 55. The method of claim 54, wherein said measuring said rate of metabolism comprises measuring the toxicity or activity mediated by the administration of said compound, measuring the half-life of said compound, or measuring the serum level of a liver enzyme.
- 56. A screening method for determining whether the metabolism of a first compound is modulated by a second compound, said method comprising the steps of:
(a) administering said first compound in the presence and absence of said second compound to a mouse, said mouse comprising a mutation that reduces CAR receptor activity; and (b) in the presence and absence of said second compound, measuring a physiological effect that is mediated by the administration of said first compound, whereby said second compound is determined to modulate the metabolism of said first compound if the change effected by said second compound in said physiological effect mediated by said administration of said first compound is smaller in said mouse comprising said mutation than in a mouse having wild-type CAR receptor activity.
- 57. The method of claim 56, wherein said measuring said physiological effect comprises measuring the toxicity or activity mediated by the administration of said first compound or measuring the half-life of said first compound.
- 58. The method of claim 57, wherein said toxicity or activity is mediated by a metabolite of said first compound.
- 59. The method of claim 57, wherein step (b) comprises measuring the half-life of said first compound in the presence and absence of said second compound, whereby said second compound is determined to activate the metabolism of said first compound if the decrease in said half-life effected by said second compound is smaller in said mouse comprising said mutation than in a mouse having wild-type CAR receptor activity, or said second compound is determined to inhibit the metabolism of said first compound if the increase in said half-life effected by said second compound is smaller in said mouse comprising said mutation than in a mouse having wild-type CAR receptor activity.
- 60. The method of claim 45, 48, 51, 54, or 56, wherein said mouse is a transgenic mouse.
- 61. The method of claim 45, 48, 51, 54, or 56, wherein said mutation substantially eliminates CAR receptor activity.
- 62. The method of claim 29, 38, 45, or 54, wherein said compound is eliminated from drug development.
- 63. The method of claim 40 or 56, wherein said first compound activates the metabolism of said second compound, and said first compound or said second compound is eliminated from drug development.
- 64. The method of claim 29, 32, 45, or 48 wherein said CAR target gene is CYP2B10 or CYP2B6.
- 65. The method of claim 29, 32, 35, 38, 40, 45, 48, 51, 54, or 56, wherein at least one of said compound, said first compound, or said second compound is a member of a library of at least 5 compounds, all of which are simultaneously administered to said mouse comprising said mutation or said mouse expressing said human CAR receptor.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of International Application PCT/US01/29672, filed Sep. 21, 2001 and published in English, and a continuation-in-part of U.S. utility application 10/219,590, filed Aug. 15, 2002, both of which claim priority to U.S. utility application Ser. No. 09/666,250, filed Sep. 21, 2000.
STATEMENT AS TO FEDERALLY SPONSORED RESEARCH
[0002] This invention was made with government support under NIH grant NIDDK RO1 DK46546. The government therefore has certain rights in the invention.
Continuation in Parts (2)
|
Number |
Date |
Country |
Parent |
PCT/US01/29672 |
Sep 2001 |
US |
Child |
10268822 |
Oct 2002 |
US |
Parent |
10219590 |
Aug 2002 |
US |
Child |
10268822 |
Oct 2002 |
US |