Alkaline phosphatase polypeptides and methods of use thereof
Inventors
Voegtli, Walter C. • BARANELLO, Michael • BOUCHARD, Keith
Assignees
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Publication Number
US-12433938-B2
Publication Date
2025-10-07
Expiration Date
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).
Core Innovation
The disclosure describes soluble alkaline phosphatase (sALP) therapeutic polypeptides and fusions intended for bone mineralization disorders, including hypophosphatasia (HPP). It focuses on recombinant alkaline phosphatase polypeptides that are engineered for altered properties and for controlling pyrophosphate (PPi) and bone mineralization outcomes.
The invention provides recombinant alkaline phosphatase polypeptides with at least one specified mutation selected from E108S, E108T, E108Q, E108M, E108K, E108L, M384R, and L385T relative to SEQ ID NO: 1. The polypeptides further include an amino acid sequence having at least 90% sequence identity to specified SEQ ID NOs, with mutation positions defined relative to the listed SEQ ID NOs.
To associate the engineered alkaline phosphatase with bone, the polypeptide includes a bone targeting moiety comprising 3 to 30 consecutive aspartate or glutamate residues. The disclosure also describes bone-targeting constructs that can be combined with additional regions such as immunoglobulin Fc regions and other fusion elements.
The disclosure describes an injectable pharmaceutical formulation comprising soluble alkaline phosphatase (sALP) or an sALP-fusion polypeptide. The formulations are clear aqueous solutions with a pH of about 7.4, and include pharmaceutically acceptable carriers such as sodium chloride and/or sodium phosphate.
Claims Coverage
The provided claim material covers recombinant alkaline phosphatase polypeptides with specified mutation options relative to SEQ ID NO: 1 and a bone targeting moiety of consecutive aspartate or glutamate residues, together with sequence-identity constraints to specified SEQ ID NOs. Additional claim refinements restrict mutation count, raise sequence-identity thresholds or specify exact sequences, constrain bone-targeting length, and define formulation parameters.
Recombinant alkaline phosphatase with specified mutations relative to SEQ ID NO: 1
A polypeptide comprising a recombinant alkaline phosphatase having at least one mutation selected from E108S, E108T, E108Q, E108M, E108K, E108L, M384R, and L385T relative to SEQ ID NO: 1.
Bone-targeting moiety of consecutive aspartate or glutamate residues
A polypeptide further comprising a bone targeting moiety comprising from 3 to 30 consecutive aspartate or glutamate residues.
High sequence identity to specified SEQ ID NOs with mutation positions referenced
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.
Multiple mutation-count constraint
The polypeptide comprises a recombinant alkaline phosphatase containing two to five mutations chosen from E108S, E108T, E108Q, E108M, E108K, E108L, M384R, and L385T relative to SEQ ID NO: 1.
Raised sequence identity threshold
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.
Exact sequence inclusion of SEQ ID NOs
The polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 72, 123, 155, or 177.
Bone-targeting moiety length constrained by Dn/En with n=7 to 10
The bone targeting moiety comprises Dn or En, wherein n=7 to 10.
Pharmaceutical composition with sodium chloride/sodium phosphate carriers and dosage/volume ranges
A pharmaceutical composition is defined by a pharmaceutically acceptable carrier containing sodium chloride and/or sodium phosphate, a polypeptide dosage ranging from 0.1 mg/ml to 10 mg/mL, and/or a formulation volume of 0.1 mL to 50 mL.
The core claimed invention is a recombinant alkaline phosphatase polypeptide with specified mutation options and a consecutive aspartate/glutamate bone targeting moiety, constrained by sequence identity to specified SEQ ID NOs. Dependent refinements include restricting the number of mutations, increasing sequence identity or specifying exact sequences, constraining bone-targeting length, and defining a pharmaceutical composition with sodium chloride and/or sodium phosphate plus dosage and volume ranges.
Stated Advantages
Improved activity and/or PK, including improved catalytic activity, stability, zinc binding, reduced aggregation, and improved Km/AUC.
Bone mineralization outcomes supported by functional endpoints and biomarker-related measurements described in the document.
Documented Applications
Treatment framework for bone mineralization disorders, including hypophosphatasia (HPP), using sALP therapeutic polypeptides and fusions.
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