Polymeric forms of H-NOX proteins
Inventors
KAPP, Gregory • SERWER, LAURA • LE MOAN, NATACHA • Cary, Stephen P. L.
Assignees
Publication Number
US-10766947-B2
Publication Date
2020-09-08
Expiration Date
2033-01-07
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Abstract
The invention provides polymeric H-NOX proteins for the delivery of oxygen with longer circulation half-lives compared to monomeric H-NOX proteins. Polymeric H-NOX proteins extravasate into and preferentially accumulate in tumor tissue for sustained delivery of oxygen. The invention also provides the use of H-NOX proteins as radiosensitizers for the treatment of brain cancers.
Core Innovation
The invention provides polymeric H-NOX proteins comprising two or more H-NOX domains for delivering oxygen with longer circulation half-lives compared to monomeric H-NOX proteins. These polymeric H-NOX proteins extravasate into and preferentially accumulate in tumor tissue for sustained oxygen delivery. The invention also discloses the use of H-NOX proteins as radiosensitizers for treating brain cancers, including glioblastoma.
H-NOX proteins are a conserved family of hemoproteins that bind nitric oxide and oxygen via a heme domain. Unlike previous hemoglobin-based oxygen carriers, H-NOX proteins are nitric-oxide-neutral, exhibiting low NO reactivity and high NO stability, reducing hypertensive or renal side effects. Monomeric H-NOX proteins are relatively small and have short circulation half-lives due to renal filtration.
To address these limitations, the invention introduces polymeric H-NOX proteins which have longer circulation half-lives, preferentially accumulate in tumors, including hypoxic regions, and are tuned to release oxygen in these hypoxic tumor zones. Polymeric H-NOX proteins are constructed by linking H-NOX domains to polymerization domains such as a trimerization foldon domain from bacteriophage T4 fibritin, resulting in dimers, trimers, tetramers, or pentamers with enhanced therapeutic properties as oxygen carriers and radiosensitizers.
Claims Coverage
The independent claim covers a method of treating diseases in mammals by administering a specific trimeric H-NOX protein. The inventive features detail the specific composition, mutations, linkages, and method of administration of this protein.
Trimeric H-NOX protein for treatment
A trimeric H-NOX protein comprising three H-NOX monomers, each monomer having a T. tengcongensis H-NOX domain with an L144F substitution and a bacteriophage T4 fibritin foldon trimerization domain is administered to treat diseases or conditions in mammals.
Disease or condition coverage
The method applies to treating cardiovascular diseases where blood volume or oxygen carrying capacity is reduced, traumatic blood loss, hemorrhagic shock, surgery involving high blood loss, hemodilution, wounds including diabetic ulcers and burn wounds, anemia, tissue ischemia including stroke, myocardial infarction, and traumatic brain injury.
PEGylation of H-NOX monomers
Each of the three H-NOX monomers of the trimeric H-NOX protein may be PEGylated to improve therapeutic properties.
Covalent linkage between domains
The C-terminus of the H-NOX domain of each monomer is covalently linked to the foldon trimerization domain, optionally via an amino acid linker such as a Gly-Ser-Gly linker of three amino acids.
Specific amino acid sequences
The monomers can comprise the amino acid sequence set forth in SEQ ID NO:8 or the T. tengcongensis H-NOX domain as SEQ ID NO:2 with L144F substitution.
Method of administration and subject
The method involves administration to a mammal, including humans, preferably by intravenous injection.
The claims cover a therapeutic method using a specifically constructed trimeric mutant H-NOX protein designed for oxygen delivery in a range of diseases associated with oxygen deficiency or blood loss. Key inventive aspects include the mutant H-NOX domain, the trimerization foldon domain, covalent linkages, specific linker sequences, and therapeutic applications for mammals, with options for PEGylation and intravenous administration.
Stated Advantages
Polymeric H-NOX proteins have longer circulation half-lives compared to monomeric H-NOX proteins.
They preferentially accumulate and persist in tumor tissues, allowing sustained delivery of oxygen.
Low nitric oxide reactivity reduces hypertensive or renal side effects associated with hemoglobin-based oxygen carriers.
Improved oxygen delivery enhances the efficacy of radiotherapy and chemotherapy by modifying tumor hypoxia.
The foldon domain promotes stable trimerization, increasing molecular weight and plasma retention.
Documented Applications
Delivery of oxygen in mammals including treatment of cardiovascular diseases with blood loss or reduced oxygen capacity, trauma, hemorrhagic shock, surgeries causing blood loss, hemodilution, wounds, diabetic ulcers, anemia, tissue ischemia, stroke, myocardial infarction, and traumatic brain injury.
Use as a radiosensitizer to treat brain cancers, including glioblastoma, enhancing radiation therapy outcomes by oxygenating hypoxic tumor regions.
Use in oncology to reduce hypoxia in solid tumors to improve chemotherapeutic and radiotherapeutic efficacy.
Therapeutic agent in veterinary medicine for treatment of pets, laboratory research animals, farm animals including dogs, cats, horses, monkeys, rabbits, rats, mice, guinea pigs, hamsters, pigs, or cows.
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