Chimeric fibroblast growth factor 19 proteins and methods of use
The present invention relates to a chimeric protein that includes an N-terminus coupled to a C-terminus, where the N-terminus includes a portion of a paracrine fibroblast growth factor (“FGF”) and the C-terminus includes a C-terminal portion of an FGF19 molecule. The portion of the paracrine FGF is modified to decrease binding affinity for heparin and/or heparan sulfate compared to the portion without the modification. The present invention also relates to pharmaceutical compositions including chimeric proteins according to the present invention, methods for treating a subject suffering from diabetes, obesity, or metabolic syndrome, and methods of screening for compounds with enhanced binding affinity for the βKlotho-FGF receptor complex involving the use of chimeric proteins of the present invention.
1. A method for decreasing blood glucose level in a subject suffering from diabetes, the method comprising:
selecting a subject suffering from diabetes;
providing a chimeric fibroblast growth factor (FGF) protein, wherein the chimeric FGF protein comprises an N-terminus coupled to a C-terminus, wherein the N-terminus comprises an FGF2 portion beginning at any one of residues 1 to 25 and ending at any one of residues 151 to 155 of SEQ ID NO: 121, wherein the FGF2 amino acid positions corresponding to those selected from the group consisting of N36, K128, R129, K134, K138, Q143, K144, and combinations thereof are substituted to decrease binding affinity for heparin and/or heparan sulfate compared to FGF2 without the substitution, and wherein the C-terminus comprises a portion of an FGF19 comprising amino acid residues 169 to 216 of SEQ ID NO: 233; and
administering to said selected subject an effective amount of the chimeric FGF protein to decrease blood glucose level in the selected subject.
2. The method according to claim 1 , wherein the FGF2 portion is amino acid residues 1-151 of SEQ ID NO: 121.
3. The method according to claim 1 , wherein the FGF2 portion is amino acid residues 25-151 of SEQ ID NO: 121.
4. The method according to claim 1 , wherein the FGF2 portion is amino acid residues 1-152, 1-153, 1-154, 1-155, 2-151, 2-152, 2-153, 2-154, 2-155, 3-151, 3-152, 3-153, 3-154, 3-155, 4-151, 4-152, 4-153, 4-154, 4-155, 5-151, 5-152, 5-153, 5-154, 5-155, 6-151, 6-152, 6-153, 6-154, 6-155, 7-151, 7-152, 7-153, 7-154, 7-155, 8-151, 8-152, 8-153, 8-154, 8-155, 9-151, 9-152, 9-153, 9-154, 9-155, 10-151, 10-152, 10-153, 10-154, 10-155, 11-151, 11-152, 11-153, 11-154, 11-155, 12-151, 12-152, 12-153, 12-154, 12-155, 13-151, 13-152, 13-153, 13-154, 13-155, 14-151, 14-152, 14-153, 14-154, 14-155, 15-151, 15-152, 15-153, 15-154, 15-155, 16-151, 16-152, 16-153, 16-154, 16-155, 17-151, 17-152, 17-153, 17-154, 17-155, 18-151, 18-152, 18-153, 18-154, 18-155, 19-151, 19-152, 19-153, 19-154, 19-155, 20-151, 20-152, 20-153, 20-154, 20-155, 21-151, 21-152, 21-153, 21-154, 21-155, 22-151, 22-152, 22-153, 22-154, 22-155, 23-151, 23-152, 23-153, 23-154, 23-155, 24-151, 24-152, 24-153, 24-154, 24-155, 25-152, 25-153, 25-154, or 25-155 of SEQ ID NO: 121.
5. The method according to claim 1 , wherein the one or more substitutions are selected from the group consisting of N36T; K128D; R129Q; K134V; K138H; Q143M; K144T, K144L, or K144I; and combinations thereof.
6. The method according to claim 1 , wherein the subject has type II diabetes, gestational diabetes, or drug-induced diabetes.
7. The method according to claim 1 , wherein the subject has type I diabetes.
8. The method according to claim 1 , wherein the subject is obese.
9. The method according to claim 1 , wherein the disorder subject has metabolic syndrome.
10. The method according to claim 1 , wherein the administering is performed subcutaneously, intravenously, intramuscularly, intraperitoneally, by intranasal instillation, by implantation, by intracavitary or intravesical instillation, intraocularly, intraarterially, intralesionally, transdermally, or by application to mucous membranes.
11. The method according to claim 1 , wherein the chimeric FGF protein is administered with a pharmaceutically-acceptable carrier.
12. The method according to claim 1 , wherein the selected subject is a mammal.
13. The method according to claim 1 , wherein the selected subject is a human.
14. The method according to claim 1 , wherein the chimeric FGF protein is co-administered with one or more agents selected from the group consisting of an anti-inflammatory agent, an antifibrotic agent, an antihypertensive agent, an antidiabetic agent, a triglyceride-lowering agent, and a cholesterol-lowering agent.
15. The method according to claim 1 , wherein the chimeric FGF protein comprises the amino acid sequence of SEQ ID NO:335 or SEQ ID NO:336.
16. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue N36.
17. The method according to claim 16 , wherein the substitution is N36T.
18. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue K128.
19. The method according to claim 18 , wherein the substitution is K128D.
20. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue R129.
21. The method according to claim 20 , wherein the substitution is R129Q.
22. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue K134.
23. The method according to claim 22 , wherein the substitution is K134V.
24. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue K138.
25. The method according to claim 24 , wherein the substitution is K138H.
26. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue Q143.
27. The method according to claim 26 , wherein the substitution is Q143M.
28. The method according to claim 1 , wherein the one or more amino acid substitutions comprises a substitution at amino acid residue K144.
29. The method according to claim 28 , wherein the substitution is K144T.
30. The method according to claim 28 , wherein the substitution is K144L.
31. The method according to claim 28 , wherein the substitution is K144I.
32. The method according to claim 1 , wherein the one or more amino acid substitutions comprises substitutions at amino acid residues K128, R129, and K134.
33. The method according to claim 32 , wherein the substitutions are K128D, R129Q, and K134V.