Human FGF23 antagonist

Abstract

The problem that this invention aims to solve is to improve hFGF23 antagonist compounds. [Solution] The present invention relates to a novel hFGF23 antagonist polypeptide, its long-acting derivatives, and its use in pharmaceuticals, and more particularly to inhibiting hFGF23-induced signaling in patients who require it, such as patients suffering from hypophosphatemic disorders like X-linked hypophosphatemic rickets (XLH), and pharmaceutical compositions containing such compounds.

Claims

1. The location on hFGF23 determined at 3.5 Å by Sequence ID No. 1 a) R76, T86, R91, and R114, b) M74, R76, T86, R91, F108, P110, R114, L166, I167, and N170, c) R76, T86, R91, Y93, Y107, R114, F169, and N170, d) W36, R76, T86, R91, Y107, R114, I167, F169, and N170, and e) R76, T86, R91, R114, L166, and F169, hFGF23 antagonist polypeptide capable of binding to an epitope containing an amino acid residue selected from the group consisting of the following.

2. hFGF23 antagonist polypeptide, which is capable of binding to hFGF23 according to SEQ ID NO: 1, i) X according to SEQ ID NO: 2 ａ X ｂ X ｃ AWX ｄ EIX ｅ X ｆ X ｇ PX ｈ LX ｉ DX ｊ QWPAFIEX ｋ LH and During the ceremony, they act independently of each other. X ａ is F or Y, X ｂ is F, I, L, or V, X ｃ is A or Q, X ｄ is A, F, H, Y, or W, X ｅ is F or Y, X ｆ is N, Q, or T, X ｇ is A or L, X ｈ is C, H, N, Q, W, or Y, X ｉ is D, N, Q, S, or T, X ｊ is A, D, E, G, H, L, N, Q, S, T, or Y, X ｋ This is an amino acid residue sequence that is A, Q, or S. and ii) An amino acid residue sequence having at least 93% identity with the sequence defined in i) hFGF23 antagonist polypeptide containing a binding motif (BM) consisting of a selected amino acid residue sequence.

3. The hFGF23 antagonist polypeptide according to claim 2, wherein (BM) is Selected from the group consisting of SEQ ID NOs: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, and 52; an hFGF23 antagonist polypeptide, optionally comprising one, two, or three substitutions, the substitutions of which may occur at any one of the positions 1, 2, 3, 6, 9, 10, 11, 13, 15, 17, and / or 25 of the selected (BM) sequence.

4. The hFGF23 antagonist polypeptide according to claim 3, wherein the substitution is a conservative substitution.

5. An hFGF23 antagonist polypeptide according to any one of claims 1 to 4, The polypeptide contains a binding motif (BM), a) X according to Sequence ID 3 １ X ２ X ３ X ４ X ５ X ６ X ７ X ８ -(BM)-X ３６ X ３７ PSQX ４１ X ４２ X ４３ LLX ４６ EARX ５０ LX ５２ X ５３ X ５４ QX ５６ X ５７ X ５８ And, In the formula, (BM) consists of the amino acid residue sequence defined in claim 2(i), During the ceremony, they act independently of each other. X １ is G, I, L, N, or V, X ２ is D, E, or Q, X ３ is D, E, G, N, or Q, X ４ is D, E, H, N, R, or T, X ５ is E, I, L, T, or V, X ６ is F, W, or Y, X ７ is A, G, H, I, L, N, P, Q, R, T, or V, X ８ is A, L, or E, X ３６ is A, D, or E, X ３７ is D or E, X ４１ is A, R, or W, X ４２ is A, D, or T, X ４３ is E, N, Q, or S, X ４６ is A, E, or K, X ５０ is Q or R, X ５２ is N or E, X ５３ is D, E, K, or T, X ５４ is A or I, X ５６ is A or C, X ５７ is K or P, X ５８ This is an amino acid residue sequence that is C, K, or E. and b) An hFGF23 antagonist polypeptide comprising an amino acid residue sequence selected from an amino acid residue sequence having at least 93% identity with the sequence defined in a).

6. The hFGF23 antagonist polypeptide according to claim 5, wherein each is independently of the other X １ is G, I, L, or V, X ２ is D or E, X ３ is D or E, X ４ is D or E, X ５ E is, X ６ F is, X ７ is I or Q, X ８ is A or L, X ａ Y is, X ｂ V is, X ｃ Q is, X ｄ Y is, X ｅ is F or Y, X ｆ is N or Q, X ｇ L is, X ｈ N is, X ｉ These are N and T, X ｊ is A, N, or Q, X ｋ S is, X ３６ is D or E, X ３７ is D or E, X ４１ W is, X ４２ A is, X ４３ N is, X ４６ is A or E, X ５０ is Q, or R, and X ５２ N is, X ５３ E is, X ５４ However, A is, X ５６ However, A is, X ５７ However, P is, X ５８ This is K, an hFGF23 antagonist polypeptide.

7. The hFGF23 antagonist polypeptide according to any one of claims 2 to 6, wherein the binding motif (BM) forms part of a 3-helix vandal protein domain, and the 3-helix vandal protein domain is a variant of protein Z derived from domain B of Staphylococcus protein A.

8. hFGF23 antagonist polypeptide, which is capable of binding to hFGF23 according to SEQ ID NO: 1, Chemical substance 197 according to Sequence ID No. 168; Chemical substance 201 according to Sequence ID No. 176; Chemical substance 204 according to Sequence ID No. 234; Chemical substance 206 according to Sequence ID No. 236; Chemical substance 207 according to Sequence ID No. 237; and An hFGF23 antagonist polypeptide having at least 93% identity with an hFGF23 antagonist polypeptide selected from the group consisting of chemical substance 208, as defined by Sequence ID No.

238.

9. The hFGF23 antagonist polypeptide according to any one of claims 1 to 8, wherein the hFGF23 polypeptide is Chemical substances 89, Chemical substance 106, Chemical substance 107, Chemical substance 111, Chemical substances 197, Chemical substance 201, Chemical substance 204, Chemical substance 206, Chemical substance 207; and hFGF23 antagonist polypeptide, selected from the group consisting of 208 chemical substances.

10. An hFGF23 antagonist polypeptide according to any one of claims 1 to 9, wherein the polypeptide is capable of binding to hFGF23 at a KD value of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, less than 1 μM, less than 0.9 μM, less than 0.8 μM, less than 0.7 μM, less than 0.6 μM, less than 0.5 μM, less than 0.4 μM, less than 0.3 μM, less than 0.2 μM, less than 0.1 μM, less than 10 nM, less than 1 nM, less than 100 μM, preferably less than 0.1 μM, such as less than 10 nM, less than 2 nM, or less than 1 nM.

11. An hFGF23 antagonist polypeptide according to any one of claims 1 to 10, including an extended portion.

12. The hFGF23 antagonist polypeptide according to claim 11, wherein the extended portion is covalently bonded to the side chain group of an amino acid residue on the polypeptide.

13. The hFGF23 antagonist polypeptide according to claim 11 or 12, wherein the extended portion comprises a C16, C17, C18, C19, C20, C21, or C22 fatty acid such as C18, C19, or C20 fatty acid.

14. The hFGF23C antagonist polypeptide according to any one of claims 11 to 13, wherein the extended portion is 【Chemistry 1】 (Chemical substance 1a); 【Chemistry 2】 (Chemical substance 1b): 【Transformation 3】 (Chemical substance 1c); 【Chemistry 4】 (Chemical substance 1d); 【Transformation 5】 (Chemical substance 1e); QRLMEDICLPRWGCLWEDDF-* (chemical substance 1f); and *-QRLMEDICLPRWGCLWEDDF (chemical substance 1f); (where * represents a binding site to either a linker (L Ｐ ), or an hFGF23 antagonist polypeptide)) hFGF23 antagonist polypeptide containing extension factor P selected from the group consisting of the following.

15. The hFGF23 antagonist polypeptide according to any one of claims 1 to 14, wherein the polypeptide derivative includes an extended portion, and the extended portion is *-NH-(CH ２ ) ２ - (O - (CH ２ )) ｋ -O-(CH ２ ) ｎ -CO-* (chemical substance 2a) (wherein k is an integer in the range of 1 to 5, and n is an integer in the range of 1 to 5); *-NH-S(O) ２ -CH ２ -CH ２ -CH ２ -CO-*; (chemical substance 2e); *-NH-CH ２ - (C) ６ H １０ )-CO-*(chemical substance 2f); *-NH-(CH) ２ ) ５ -CO-* (2g of chemical substance); 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 Linker L selected from the group consisting of the following Ｐ hFGF23 antagonist polypeptide, including

16. The hFGF23 antagonist polypeptide according to claim 15, wherein the extended portion "P" is a chemical substance 1c and linker L Ｐ However, the chemical substance 2i (L1) is the hFGF23 antagonist polypeptide.

17. The hFGF23 antagonist polypeptide according to any one of claims 1 to 16, wherein the polypeptide derivative includes an extended portion, and the extended portion is covalently bonded to the side chain of a cysteine ​​or lysine residue in the polypeptide.

18. It is a compound, 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 A compound selected from the group consisting of the following.

19. A compound according to claim 18, which is an hFGF23 antagonist polypeptide derivative that can bind to hFGF23 according to SEQ ID NO:

1.

20. The hFGF23 antagonist polypeptide derivative is represented by the following formula. 【Chemistry 14】

21. A pharmaceutical composition comprising an hFGF23 antagonist polypeptide, polypeptide derivative, or compound according to any one of claims 1 to 20, and one or more pharmaceutically acceptable excipients.

22. An hFGF23 antagonist polypeptide derivative, polypeptide derivative, compound, or composition according to any one of claims 1 to 21, for use in pharmaceuticals.

23. An hFGF23 antagonist polypeptide, polypeptide derivative, compound, or composition according to any one of claims 1 to 22, for use in the treatment of tumor-induced bone disease, fibrous dysplasia, McCune-Albright syndrome, autosomal dominant hypophosphatemic rickets (ADHR), or X-linked hypophosphatemic rickets (XLH).

24. A kit comprising an hFGF23 antagonist polypeptide, polypeptide derivative, compound, or composition according to any one of claims 1 to 21, and instructions for use.

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