Formulations of human tissue kallikrein-1 for parenteral delivery and related methods
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Publication Number
US-9616015-B2
Publication Date
2017-04-11
Expiration Date
Abstract
Provided are high concentration compositions of tissue kallikrein-1 (KLK1) and methods of parenterally administering such compositions to a subject in need thereof, where absorption into the circulation via, for example, intravenous or subcutaneous administration improves systemic pharmacokinetics, bioavailability, safety, and/or convenience relative to intravenous or other forms of administration. Also provided are recombinant human KLK1 (rhKLK1) polypeptides that can be readily concentrated to high protein concentrations, and substantially pure compositions thereof.
Core Innovation
The invention relates to compositions and methods for treating a subject in need thereof by subcutaneously administering a parenteral formulation comprising a recombinant mature human tissue kallikrein-1 (hKLK1) polypeptide and a pharmaceutically acceptable carrier. The recombinant mature hKLK1 polypeptide is isolated from recombinant mammalian cells, has a pI of less than about 5, and has a sialic acid content of at least about 4 moles per mole protein.
The composition is substantially free of aggregates, with greater than about 95% appearing as a single peak by SEC HPLC, and has low levels of endotoxin, host cell protein, and host cell DNA. The polypeptide is characterized for serine-protease activity, including kallidin release (lysyl-bradykinin release).
Subcutaneous administration produces improved systemic pharmacokinetics relative to intravenous administration of the same composition. The disclosed rationale connects increased sialylation with a low pI and reduced aggregation at high concentration, enabling stable high-concentration formulations suitable for smaller-volume parenteral administration, with pharmacokinetic/toxicokinetic assessment comparing subcutaneous versus intravenous dosing.
Claims Coverage
The claim coverage centers on one independent claim covering a subcutaneous treatment method with a specifically characterized recombinant mature hKLK1 composition, and dependent claims that further refine pharmacokinetic outcomes and hKLK1 sequence/function constraints. The independent claim contains the inventive core of the formulation attributes, aggregate and impurity controls, and the subcutaneous pharmacokinetic improvement relative to intravenous administration.
Subcutaneous treatment using recombinant mature hKLK1 composition with defined pI, sialic acid, and aggregate/impurity specifications
Subcutaneously administering to the subject a composition formulated for parenteral administration, comprising a recombinant mature human tissue kallikrein-1 (hKLK1) polypeptide isolated from recombinant mammalian cells and a pharmaceutically acceptable carrier, where the mature hKLK1 polypeptide has a pI of less than about 5 and a sialic acid content of at least about 4 moles per mole protein, where the composition is substantially free of aggregates (greater than about 95% appearing as a single peak by SEC HPLC) and has endotoxin levels of less than about 1 EU/mg protein, host cell protein of less than about 100 ng/mg protein, and host cell DNA of less than about 10 pg/mg protein.
Subcutaneous administration produces improved systemic pharmacokinetics relative to intravenous administration
Where administering the composition subcutaneously produces improved systemic pharmacokinetics relative to intravenous administration of the composition, thereby treating the subject.
Improved pharmacokinetics includes an increased apparent half-life
The improved pharmacokinetics includes an increased apparent half-life.
Improved pharmacokinetics includes a decreased Tmax
The improved pharmacokinetics includes a decreased Tmax.
Improved pharmacokinetics includes an increased absorption rate
The improved pharmacokinetics includes an increased absorption rate.
Recombinant mature hKLK1 meeting sequence identity to residues 25-262 with serine protease activity
Using an hKLK1 polypeptide that has at least about 95% sequence identity to residues 25-262 of SEQ ID NO:1 or SEQ ID NO:2 and exhibits serine protease activity.
hKLK1 polypeptide consisting of residues 25-262 of SEQ ID NO:1
A hKLK1 polypeptide consisting of residues 25-262 of SEQ ID NO:1.
The claim set covers subcutaneous administration of a recombinant mature hKLK1 composition with specified pI and sialic acid, substantially free of aggregates by SEC HPLC, and low endotoxin, host cell protein, and host cell DNA, together with a requirement that the subcutaneous route yields improved systemic pharmacokinetics compared to intravenous administration. Dependent claims further narrow the pharmacokinetic improvement and the hKLK1 polypeptide identity to defined residues and serine protease activity.
Stated Advantages
Improved systemic pharmacokinetics relative to intravenous administration.
Subcutaneous administration enables treatment of a subject in need thereof via the specified recombinant hKLK1 formulation.
The composition is substantially free of aggregates (>95% single peak by SEC HPLC).
Documented Applications
Treating a subject in need thereof using subcutaneous administration of a parenteral composition comprising recombinant mature human tissue kallikrein-1 (hKLK1) with defined pI and sialic acid and low aggregate and impurity levels.
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