Nuclear receptor ligands and ligand binding domains

• US 6,236,946 B1
• Filed: 12/13/1996
• Issued: 05/22/2001
• Est. Priority Date: 12/13/1995
• Status: Expired due to Term

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Shiau, Andrew K., West, Brian, Scanlan, Thomas S., Baxter, John D., Wagner, Richard L., Fletterick, Robert J., Kushner, Peter J., Apriletti, James
• Primary Examiner(s): Eyler, Yvonne
• Assistant Examiner(s): Basi, Nirmal S.
• Application Number:
  US08/764,870
• Time in Patent Office: 1,621 Days
• Field of Search: 702/14, 702/21, 702/22, 530/350
• US Class Current: 702/22
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: C07K 14/72 for hormones for neuromediators C07K14/70571
• CPC Class Codes: C07K 2299/00 Coordinates from 3D structures of peptides, e.g. proteins or enzymes

Abstract

The present invention provides new methods, particularly computational methods, and compositions for the generation of nuclear receptor synthetic ligands based on the three dimensional structure of nuclear receptors, particularly the thyroid receptor (herein referred to as “TR”). Also provided are crystals, nuclear receptor synthetic ligands, and related methods.

Claims

1. A method of designing a nuclear receptor synthetic ligand comprising:

1) generating a three dimensional model of a protein comprising a nuclear receptor ligand binding domain (LBD) with a bound ligand utilizing data from FIGS. 28, 29, 30, or 31 and a computer programmed for generating said model from said data;

2) determining at least one interacting amino acid of said nuclear receptor LBD that interacts with at least one first chemical moiety of said bound ligand in said three dimensional model;

3) selecting at least one chemical modification of said first chemical moiety to produce a second chemical moiety;

4) measuring the reduction or enhancement of the interaction between said interacting amino acid and said second chemical moiety compared to the interaction between said interacting amino acid and said first chemical moiety;

5) generating a designed nuclear receptor synthetic ligand wherein said first chemical moiety is replaced with said second chemical moiety.

2. The method of claim 1, wherein steps 2, 3 and 4 are repeated.

3. The method of claim 1, wherein said three dimensional model comprises a thyroid hormone receptor (TR) LBD with a bound TR ligand.

4. The method of claim 3, wherein said selecting uses said first chemical moiety that interacts with at least one of the said interacting amino acids listed in FIG. 27.

5. The method of claim 4, wherein said enhancement of the interaction is enhancement of hydrogen bonding interaction, charge interaction, hydrophobic interaction, Van Der Waals interaction or dipole interaction.

6. The method of claim 4, wherein said reduction of the interaction is reduction of hydrogen bonding interaction, charge interaction, hydrophobic interaction, Van Der Waals interaction or dipole interaction.

7. The method of claim 5 wherein said measuring and replacing are performed using a computer program to represent chemical structures of said interacting amino acid and ligand.

8. The method of claim 6, wherein said TR ligand is a thyronine derivative of the formula

and said chemical modification is at the R5′ position of said thyronine derivative.

9. The method of claim 1 wherein said protein is a nuclear receptor other than TR.

10. A method of designing a nuclear receptor antagonist from a nuclear receptor agonist comprising:

1) determining a structure of a molecular recognition domain of said agonist using a three dimensional model of a protein comprising a nuclear receptor ligand binding domain (LBD) generated from data from FIGS. 28, 29, 30, or 31 and a computer programmed for generating said model when supplied with said data as input data;

2) selecting at least one chemical modification of a first chemical moiety to produce a second chemical moiety that extends beyond a binding site for said agonist and in the direction of at least one protein domain selected from the group consisting of a transcription activation domain of said LBD, a repressor binding domain of said LBD, a DNA binding domain of said nuclear receptor, a heat shock protein binding domain of said nuclear receptor, a dimerization domain of said LBD, and a hinge region to said DNA binding domain; and

3) generating a designed nuclear receptor antagonist wherein said first chemical moiety is replaced with said second chemical moiety.

11. The method of claim 10, wherein said protein comprises said nuclear receptor LBD bound to a nuclear receptor ligand.

12. The method of claim 11, wherein said nuclear receptor LBD bound to a nuclear receptor ligand comprises a thyroid hormone receptor (TR) LBD with a bound TR ligand.

13. The method of claim 12, wherein said bound TR ligand is a thyronine derivative of the formula:

14. The method of claim 11, wherein said LBD is from a receptor selected from a group consisting of glucocorticoid receptor, estrogen receptor, retinoid receptor and vitamin D receptor.

15. A method of designing a nuclear receptor super agonist or antagonist comprising:

1) generating a three dimensional model of a protein comprising a nuclear receptor ligand binding domain (LBD) with a bound ligand utilizing data from FIGS. 28, 29, 30, or 31 and a computer programmed for generating said model from said data;

2) determining at least one interacting amino acid of said nuclear receptor LBD that interacts with at least one first chemical moiety of said ligand using said three dimensional model;

3) selecting at least one chemical modification of said first chemical moiety to produce a second chemical moiety;

4) measuring the reduction or enhancement of the interaction between said interacting amino acid and said second chemical moiety compared to the interaction between said interacting amino acid and said first chemical moiety; and

5) generating a designed nuclear receptor super agonist or antagonist wherein said first chemical moiety is replaced with said second chemical moiety.

16. The method of claim 15, wherein said enhancement of the interaction is enhancement of hydrogen bonding interaction, electrostatic interaction, charge interaction, hydrophobic interaction, Van Der Waals interaction or dipole interaction.

17. The method of claim 16, wherein said chemical modification changes a carboxylate moiety of said first chemical moiety to a chemical moiety selected from the group consisting of phosphonate and phosphate.

18. The method of claim 17, wherein said nuclear receptor is a thyroid hormone receptor (TR) and said chemical modification enhances said interaction between said second chemical moiety and at least one of the following arginines:

Arg 262, Arg 266 or Arg 228 of the rat α

-TR (SEQ ID NO;

     1) or an arginine of human α

-TR or β

-TR that corresponds in its three dimensional position in said three dimensional model to said arginines;

Arg 262, Arg 266 or Arg 228.

19. A method of designing a nuclear receptor ligand comprising:

1) providing atomic coordinate data of FIGS. 28, 29, 30, or 31 to a computer having a computer program capable of generating an atomic model of a molecule from atomic coordinate data of said molecule;

2) generating with said computer an atomic model of a protein comprising a nuclear receptor ligand binding domain (LBD) or portion thereof with a bound ligand;

3) determining a first chemical moiety of said bound ligand that interacts with an amino acid of said LBD;

4) selecting at least one chemical modification of said first chemical moiety to produce a second chemical moiety;

5) measuring the reduction or enhancement of the interaction between said interacting amino acid and said second chemical moiety compared to the interaction between said interacting amino acid and said first chemical moiety; and

6) generating a designed nuclear receptor ligand having altered interaction with said amino acid of said LBD wherein said first chemical moiety is replaced with said second chemical moiety.

20. The method of claim 19, wherein said bound ligand comprises the formula:

21. The method of claim 19, wherein said nuclear receptor LBD comprises a thyroid hormone receptor (TR) LBD.

22. The method of claim 19, wherein said nuclear receptor ligand comprises the formula: