Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-10717970-B2
Publication Date
2020-07-21
Expiration Date
Abstract
Short-acting Factor VII peptides are disclosed. A shortened half-life is desirable for treatment of acute bleeding and similar disorders. Modification of the sialylation and/or glycosylation of Factor VII and variants thereof produced peptides useful in treating conditions of acute bleeding.
Core Innovation
The invention relates to short-acting Factor VII polypeptides comprising wild-type or variant amino acid sequences having at least 95% identity to the amino acid sequence of SEQ ID NO: 16. The polypeptides are characterized by reduced sialic acid conjugation and/or altered N-glycosylation in order to shorten in vivo half-life and improve safety and therapeutic window for acute bleeding.
One described core strategy is desialylation, including enzymatic desialylation and/or expression in sialylation-deficient cell lines to produce polypeptides with reduced conjugated sialic acid. Another described core strategy is deglycosylation by genetic disruption of N-glycosylation sites, including disruption of N-linked glycosylation sites such as N145 and N322 and additional N-glycosylation sites beyond wild-type.
The document further describes that the polypeptides retain Factor VII activity while exhibiting shortened pharmacokinetics and reduced thrombin generation and thrombus formation compared with an example parental control. The glyco-sialylation state is supported by quantitative assessment using LC-MS/DMB-based sialic acid quantification, and functional activity is supported using in vitro assays including phospholipid-dependent FX activation and thrombin generation/PL-dependent assays.
Claims Coverage
The partial content contains two independent claims directed to isolated variant Factor VII polypeptides. Across the independent claims, the coverage includes at least 95% identity to SEQ ID NO: 16, specified N-linked glycosylation sites corresponding to residues 145 and 322, and a quantitative constraint on conjugated sialic acid to N-linked glycan ratio between 0 and 1.0, with one independent claim additionally requiring the P10Q and K32E alterations and one requiring additional N-linked glycosylation site(s) beyond wild-type.
Variant Factor VII polypeptide with controlled glyco-sialylation and additional N-linked glycosylation sites
An isolated variant Factor VII polypeptide with an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 16, comprising the N-linked glycosylation sites of wild-type human Factor VII at amino acid residues 145 and 322 and further comprising additional N-linked glycosylation site(s) beyond the two present in wild-type Factor VII of SEQ ID NO: 16, wherein the polypeptide has a ratio of moles of conjugated sialic acid to moles of N-linked glycan between 0 and 1.0.
Defined P10Q and K32E Factor VII variant with controlled glyco-sialylation
An isolated variant Factor VII polypeptide comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 16 and comprising the sequence alterations P10Q and K32E, wherein the variant comprises N-linked glycosylation sites of wild-type human Factor VII at amino acid residues 145 and 322, and wherein the polypeptide has a ratio of moles of conjugated sialic acid to moles of N-linked glycan between 0 and 1.0.
Overall, the independent-claim coverage is directed to isolated Factor VII variants defined by SEQ ID NO: 16 identity, specified N-linked glycosylation at residues 145 and 322, a quantitative conjugated-sialic-acid-to-N-linked-glycan ratio between 0 and 1.0, and in one claim additional P10Q and K32E alterations.
Stated Advantages
Shortens in vivo half-life for acute bleeding.
Improves safety and therapeutic window for acute bleeding.
Produces reduced thrombin generation and reduced thrombus formation compared with a parental control while retaining clotting activity.
Documented Applications
Treatment of disease or disorder where blood clot formation is desirable by administering an effective amount of the isolated variant Factor VII polypeptide to a mammal, including bleeding conditions such as hemorrhage, gastrointestinal bleeding, uncontrolled bleeding, bleeding during transplantation or resection or surgery, variceal bleeding, thrombocytopenia, hemophilia, intracranial hemorrhage, aortic aneurysm, traumatic injuries, and reversal scenarios of warfarin, heparin, and other anticoagulants and antithrombotics.
Prophylaxis and treatment contexts including prophylaxis in hemophilia patients with inhibitors, bleeding in stem cell transplantation and liver transplantation, active variceal bleeding in cirrhosis, non variceal bleeding in cirrhosis, diffuse alveolar hemorrhage, intracerebral hemorrhage, traumatic brain injury, brain contusion, acquired hemophilia, idiopathic thrombocytopenic purpura, Glanzmann’s Thrombasthenia and refractory Glanzmann’s Thrombasthenia to platelet transfusion, and Bernard-Soulier Syndrome.
Interested in licensing this patent?