Alkaline phosphatase polypeptides and methods of use thereof

Patent metadata

Field	Value
Publication number	US-12433938-B2
Application number	US-202017117099-A
Country	US
Kind code	B2
Filing date	Dec 9, 2020
Priority date	Dec 9, 2019
Publication date	Oct 7, 2025
Grant date	Oct 7, 2025

Abstract

Featured are polypeptides that include soluble alkaline phosphatases, mutants, fragments, fusion proteins thereof, and methods of use thereof, for treating bone mineralization disorders, such as hypophosphatasia (HPP), and symptoms thereof. The polypeptides include a soluble alkaline phosphatase (sALP) or fragment thereof, which is derived from a naturally occurring alkaline phosphatase (ALP).

First claim

The invention claimed is:

1. A polypeptide comprising: (1) a recombinant alkaline phosphatase having at least one mutation selected from the group consisting of E108S, E108T, E108Q, E108M, E108K, E108L, M384R, and L385T relative to SEQ ID NO: 1 and (2) a bone targeting moiety comprising from 3 to 30 consecutive aspartate or glutamate residues, wherein the polypeptide comprises an amino acid sequence with at least 90% sequence identity to any one of SEQ ID NOs: 72, 123, 155, or 177, wherein the original residue before mutation is shown in SEQ ID NO: 1, and the position numbers are relative to SEQ ID NOs: 1, 72, 123, 155, and 177.

2. The polypeptide of claim 1 , wherein the polypeptide further comprises a fragment crystallizable (Fc) region of an immunoglobulin.

3. The polypeptide of claim 2 , wherein the Fc region comprises IgG1, IgG2, IgG3, or IgG4, or a chimera thereof.

4. The polypeptide of claim 3 , wherein the Fc region comprises an IgG2/4 chimera.

5. The polypeptide of claim 4 , wherein the Fc region comprises the sequence of SEQ ID NO: 253 or has at least 85% sequence identity thereto.

6. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to any one of SEQ ID NOs: 72, 123, 155, or 177.

7. The polypeptide of claim 1 , wherein the bone targeting moiety comprises Dn or En, wherein n=7 to 10.

8. The polypeptide of claim 7 , wherein the bone targeting moiety is
Dn and n=7 to 10.

9. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.

10. The pharmaceutical composition of claim 9 , wherein:
(a) the pharmaceutically acceptable carrier comprises sodium chloride and/or sodium phosphate;
(b) the composition comprises the polypeptide at a dosage of from 0.1 mg/ml to 10 mg/mL; and/or
(c) the composition is formulated in a volume of 0.1 mL to 50 mL.

11. The polypeptide of claim 1 , wherein the recombinant alkaline phosphatase has at least two, three, four, or five mutations selected from the group consisting of E108S, E108T, E108Q, E108M, E108K, E108L, M384R, and L385T relative to SEQ ID NO: 1.

12. The polypeptide of claim 6 , wherein the polypeptide comprises an amino acid sequence having at least 97% sequence identity to any one of SEQ ID NOs: 72, 123, 155, or 177.

13. The polypeptide of claim 12 , wherein the polypeptide comprises an amino acid sequence having at least 99% sequence identity to any one of SEQ ID NOs: 72, 123, 155, or 177.

14. The polypeptide of claim 13 , wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 72, 123, 155, or 177.

15. The polypeptide of claim 14 , wherein the polypeptide consists of the amino acid sequence of any one of SEQ ID NOs: 72, 123, 155, or 177.

16. The polypeptide of claim 14 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 123.

17. The polypeptide of claim 16 , wherein the polypeptide consists of the amino acid sequence of SEQ ID NO: 123.

Assignees

• Alexion Pharma Inc

Inventors

• Voegtli Walter C
• Baranello Michael
• Bouchard Keith

Classifications

• A61P19/08 Primary

  for bone diseases, e.g. rachitism, Paget's disease · CPC title

• C12Y301/03001

  Alkaline phosphatase (3.1.3.1) · CPC title

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title