US-12616742-B2 - Recombinant heme oxygenase-1 (HO-1) for the treatment of sickle cell disease

US12616742B2US 12616742 B2US12616742 B2US 12616742B2US-12616742-B2

Abstract

The present invention provides, among other things, methods and compositions for making and using recombinant heme oxygenase for treating sickle cell disease. In some embodiments, recombinant heme oxygenase proteins are truncation variants, or Fc fusion proteins with increased half-life and/or reduced aggregation.

Inventors

Solomon Fiifi Ofori-Acquah
Samit Ghosh
Joseph Sypek
Bohong Zhang
Xiuxia Sun
Clark Q. Pan
Daniel Mainard Lajoie
Chuan Shen

Assignees

TAKEDA PHARMACEUTICAL COMPANY LIMITED
UNIVERSITY OF PITTSBURGH-OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION

Dates

Publication Date: 20260505
Application Date: 20200724

Claims (5)

1 . A method of treating sickle cell disease comprising administering to a subject in need of treatment a truncated recombinant heme oxygenase-1 (rHO-1) protein, comprising an amino acid sequence with at least 85% identity to residues 1-261 of SEQ ID NO: 1, wherein the rHO-1 protein comprises an F33L amino acid substitution in SEQ ID NO: 1.
2 . The method of claim 1 , wherein the rHO-1 protein comprises an Fc domain fused to the rHO-1 protein domain, wherein the Fc domain is fused to the N-terminus or C-terminus of the rHO-1 protein domain.
3 . The method of claim 2 , wherein the rHO-1 protein is a multimer comprising at least one monomer comprising an Fc domain fused to an rHO-1 protein domain.
4 . The method of claim 1 , wherein the rHO-1 protein is truncated at T261.
5 . The method of claim 1 , wherein the rHO-1 protein comprises an amino acid sequence with 95% identity to residues 1-226 of SEQ ID NO: 1.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application is a 35 U.S.C. § 371 National Stage Application of International Application No. PCT/US20/43452, filed on Jul. 24, 2020, which claims priority to U.S. Patent Application Ser. Nos. 62/879,131 filed Jul. 26, 2019; and 63/049,285 filed Jul. 8, 2020; the entirety of each of which is hereby incorporated by reference. STATEMENT OF FEDERALLY FUNDED RESEARCH This invention was made with United States government support under grant U01HL117721 awarded by the National Institutes of Health. The United States government has certain rights in the invention. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 17, 2020, is named SHR-2009WO_SL.txt and is 200,037 bytes in size. BACKGROUND Sickle cell disease (SCD) is a chronic life-threatening blood disorder that is inherited as an autosomal recessive trait. SCD is associated with many acute and chronic complications resulting in part from an excess amount of cell-free hemoglobin (Hb) and cell-free heme due to severe hemolysis. Patients with SCD have sickle-shaped red blood cells that get lodged in small blood vessels, obstructing the flow of blood and oxygen to major organs in the body. Such blockages result in severe pain, organ damage, stroke and other complications including increased vulnerability to infection, fatigue, and delayed growth. Most people with the disease have shortened life spans. There is a need for effective therapies to treat patients with complications arising from SCD. SUMMARY OF THE INVENTION The present invention provides an effective method for treating SCD. The present invention is based, in part, on the discovery that systemic administration of a recombinant heme oxygenase (e.g., a truncated HO-1 protein or a HO-1-Fc fusion protein) reduces or ameliorates symptoms of sickle cell disease in a SCD mouse model. Without wishing to be bound by any particular theory, it is contemplated that HO-1 specifically targets and degrades free heme by converting cell free heme into cytoprotective/anti-inflammatory by-products. Administration of a recombinant heme oxygenase augments the HO-1 activity in plasma, reduces anemia, prevents acute chest syndrome (ACS), pulmonary hypertension, and acute damage to the lungs in a SCD disease model. Furthermore, the SCD therapy described herein harnesses the physiological specificity that an endogenous HO-1 protein has and can potentially minimize off-target effects. Described herein are recombinant therapeutic HO-1 proteins which retain enzymatic activity while extending half-life, increasing stability, and decreasing aggregation compared to a naturally-occurring HO-1 protein. In one aspect, the present invention provides a method of treating sickle cell disease comprising administering to a subject in need of treatment a recombinant heme oxygenase-1 (rHO-1) protein. In some embodiments, the method comprises administering an rHO-1 protein comprising an amino acid sequence with at least 85% identity to residues 1-261 of SEQ ID NO:1 (wild type full length rHO-1 protein). In some embodiments, the method comprises administering an rHO-1 protein comprising an amino acid sequence with at least 90% identity to residues 1-261 of SEQ ID NO:1 (wild type full length rHO-1 protein). In some embodiments, the method comprises administering an rHO-1 protein comprising an amino acid sequence with at least 95% identity to residues 1-261 of SEQ ID NO:1 (wild type full length rHO-1 protein). In some embodiments, the method comprises administering an rHO-1 protein comprising an amino acid sequence identical to residues 1-261 of SEQ ID NO:1 (wild type full length rHO-1 protein). In some embodiments, the method comprises administering an rHO-1 protein comprising SEQ ID NO:1. In some embodiments, the method comprises administering an rHO-1 protein comprising K18, T21, H25, Y134, G143, L147, K179, and F207 (corresponding to amino acids positions of SEQ ID NO:1). In some embodiments, the method comprises administering an rHO-1 protein wherein the rHO-1 protein is truncated at the N-terminus at a residue corresponding to M9 of SEQ ID NO: 1. In some embodiments, the method comprises administering an rHO-1 protein, wherein the rHO-1 protein comprises residues 10-225 of SEQ ID NO: 1. In some embodiments, the method comprises administering an rHO-1 protein, wherein the rHO-1 protein comprises residues 10-226 of SEQ ID NO: 1. In some embodiments, the method comprises administering an rHO-1 protein, wherein the rHO-1 protein comprises residues 10-261 of SEQ ID NO: 1. In some embodiments, the method comprises administering an rHO-1 protein wherein the rHO-1 protein is truncated at residues corresponding to K226, A233, R237, T261, and/or A265. In some embodiments, the method comprises administering an rHO-1 protein wherein the rHO-1 prote