Abstract: The application discloses compounds useful in treatment of diabetes, weight loss and/or reduction of cardiovascular risks. The compounds are bi-functional and therefore suitable as a simple treatment for patients that may benefit from treatment with both a GLP-1 receptor agonist and a PCSK9 inhibitor.
We Claim
1. A compound comprising a GLP-1 analogue and an EGF(A) analogue, wherein
i. said GLP-1 analogue has at most 6 amino acid substitutions compared to GLP-1(7-37) identified by SEQ ID NO: 137 and ii. said EGF(A) analogue has 1-8 amino acid substitutions compared to the EGF(A) domain of LDL-R (293-332) identified by SEQ ID No:1, and comprises 301Leu and wherein the compound is a GLP-1 agonist and has a Ki below 8 nM when measured in the PCSK9-LDL-R binding competitive ELISA assay as described in Section C3.
2. The compound according to claim 1, wherein the compound comprises a fusion polypeptide comprising the GLP-1 analogue and the EGF(A) analogue.
3. The compound according to claim 2, wherein the fusion polypeptide comprises the GLP-1 analogue in the N-terminal and the EGF(A) analogue in the C-terminal.
4. The compound according to any of the previous claims, wherein the GLP-1 analogue comprises one or two Lys residues selected from the group consisting of: 12K, 21K, 23K, 24K, 25K, 26K, 27K, 30K, 31K, 32K, 33K, 34K and 36K.
5. The compound according to any of the previous claims, wherein the GLP-1 analogue is selected from the group of GLP-1 analogues defined by SEQ ID NO’s 138 to 187.
6. The compound according to any of the previous claims, wherein the compound has an EC50 in the GLP-1 in vitro potency assay described in C1 without HSA of at most 100 pM.
7. The compound according to any of the previous claims, wherein the GLP-1 analogue comprises 8Aib.
8. The compound according to any of the previous claims, wherein the GLP-1 analogue comprises 34R or 34Q.
9. The compound according to any of the previous claims, wherein GLP-1 analogue comprises 8Aib and 34R.
10. The compound according to any of the previous claims, wherein the GLP-1 analogue is selected from the group of GLP-1 analogues defined by SEQ ID NO.: 139 and 147-154 and 184-186.
11. The compound according to any of the previous claims, wherein the GLP-1 analogue is defined by SEQ ID NO.: 139.
12. The compound according to any of the previous claims, wherein the EGF(A) analogue comprises 309R.
13. The compound according to any of the previous claims, wherein the EGF(A) analogue comprises 321D or 321E.
14. The compound according to any of the previous claims, wherein the EGF(A) analogue comprises 312E, 312D, 312Q or 312R.
15. The compound according to any of the previous claims, wherein the EGF(A) analogue is selected from the group of EGF(A) analogues defined by SEQ ID NOs: 19, 21, 73, 107, 108, 109, 110, 111, 112,113 and 114, such as by SEQ ID NOs: 107 and 108.
16. The compound according to any of the previous claims, wherein the EGF(A) analogue is defined by SEQ ID NO.: 108.
17. The compound according to any of the previous claims wherein the fusion polypeptide comprises a peptide spacer, such as a spacer selected from the group of spacers defined by SEQ ID NO. 115-136, such as from SEQ ID NO. 116 and 119.
18. The compound according to any of the previous claims, wherein the compound has a ratio of the apparent EGF(A) Ki measured as described in C3 and the GLP-1 potency measured as the EC50 in the in vitro potency assay described in C1 without HSA of at most 200.
19. The compound according to any of the previous claims, wherein the compound comprises at least one half-life extending substituent, such as a substituent selected from the group of substituents consisting of:
substituent #1 defined by Chem 6b, Chem 6b
substituent #2 defined by Chem 7b, Chem 7b
substituent #3 defined by Chem 8b, Chem 8b
substituent #4 defined by Chem 9b, Chem 9b
substituent #5 defined by Chem 10b, Chem 10b
substituent #6 defined by Chem 11b, Chem 11b
substituent #7 defined by Chem 12b, Chem 12b
substituent #8 defined by Chem 13b, Chem 13b
substituent #9 defined by Chem 14b, Chem 14b
substituent #10 defined by Chem 15b, Chem 15b
substituent #11 defined by Chem 16b, Chem 16b
substituent #12 defined by Chem 17b, Chem 17b
substituent #13 defined by Chem 18b, Chem 18b
substituent #14 defined by Chem 19b, Chem 19b
20. A compound selected from the group of compounds defined as GLP-1/EGF(A) compounds #41, #48, #69 and #306, such as #306 and #69, or such as #306 or #69 wherein
compound #41 comprises a fusion peptide defined by SEQ ID NO.: 190 consisting of the GLP-1 analogue [8Aib, 34R]GLP-1(7-37) defined by SEQ ID NO.: 139, the Spacer defined by SEQ ID NO.: 116, and the EGF(A) analogue [301L, 309R, 312E, 321E, 333K]EGF(A) defined by SEQ ID NO.: 19 and two substituents defined by Chem 6b Chem 6b
attached to amino acid residues 26K of [8Aib, 34R]GLP-1(7-37) and 333K of [301L, 309R, 312E, 321E, 333K]EGF(A),
compound #48 comprises a fusion peptide defined by SEQ ID NO.: 202 consisting of the GLP-1 analogue [8Aib, 34R]GLP-1(7-37) defined by SEQ ID NO.: 139, the Spacer defined by SEQ ID NO.: 119, and the EGF(A) analogue defined by SEQ ID NO.: 108 and one substituent defined by Chem 11b Chem 11b
attached to amino acid 26K of [8Aib, 34R]GLP-1(7-37),
compound #69 comprises a fusion peptide defined by SEQ ID NO.: 310 consisting of the GLP-1 analogue [8Aib, 30G, 34R]GLP-1(7-37) defined by SEQ ID NO.: 164, the Spacer defined by SEQ ID NO.: 116, and the EGF(A) analogue defined by SEQ ID NO.: 108 and one substituent defined by Chem 11b Chem 11b
attached to amino acid 26K of [8Aib, 30G, 34R]GLP-1(7-37),
compound #306 comprises a fusion peptide defined by SEQ ID NO.: 310 consisting of the GLP-1 analogue [8Aib, 30G, 34R]GLP-1(7-37) defined by SEQ ID NO.: 164, the Spacer defined by SEQ ID NO.: 116, and the EGF(A) analogue defined by SEQ ID NO.: 108 and one substituent defined by Chem 10b Chem 10b
attached to amino acid 26K of [8Aib, 30G, 34R]GLP-1(7-37).
21. A compound according to claim 1, wherein the compound is a GLP-1/EGF(A) compound
wherein the GLP-1 analogue is defined by SEQ ID NO.: 139 and the EGF(A) analogue is
defined by SEQ ID NO.: 108.
22. A GLP-1/EGF(A) compound, wherein the compound comprises a fusion peptide defined by
SEQ ID NO. 193 and one substituent defined by Chem 6b
Chem 6b
attached via Lysine (K) in position 20 of SEQ ID 193 (equal to amino acid 26K of [8Aib, 34R]GLP-1(7-37)).
23. A compound according to any of the previous claims for use in a method of treatment of diabetes, over-weight and/or cardiovascular diseases.
24. A method of treatment of diabetes, over-weight and/or cardiovascular diseases comprising administering a pharmaceutically effective dosage of a compound according to any of claims 1-23 to a patient in need thereof.