Generation of hemoglobin-based oxygen carriers using elastin-like polypeptides

Patent metadata

Field	Value
Publication number	US-11124559-B2
Application number	US-201816206896-A
Country	US
Kind code	B2
Filing date	Nov 30, 2018
Priority date	Dec 10, 2014
Publication date	Sep 21, 2021
Grant date	Sep 21, 2021

Abstract

Described herein is the use of elastin-like polypeptides to generate hemoglobin-based oxygen carriers as a means of preventing and treating conditions caused by blood loss or anemia, for example, hemorrhagic shock. Elastin-like polypeptides are capable of creating therapeutically functional fusion proteins through genetic engineering with a therapeutic agent, for example, hemoglobin and biologic equivalent thereof. Specific forms of these fusion proteins have the ability to form into spherical nanoparticles possessing a therapeutically agent at their core. This provides a unique basis for employing elastin-like polypeptides as hemoglobin carriers in the manufacture of blood substitutes.

First claim

What is claimed is:

1. A biocompatible pharmaceutical composition comprising a therapeutic amount of a complex comprising an elastin-like polypeptide (ELP) operatively linked via fusion to a hemoglobin (Hb), a Hb subunit(s), a Hb fragment(s), or a Hb derivative(s) comprising the amino acid sequence selected from SEQ ID NO:4, SEQ ID NO: 5 or SEQ ID NO:. 6, that stores and releases oxygen in accordance with an oxygen dissociation curve; wherein the therapeutic amount of the complex is effective to treat a condition caused by blood loss, anemia, or a hemoglobin disorder, and to improve subject survival relative to a control, wherein the ELP comprises:
a hydrophilic block comprising a pentameric amino acid motif (Val-Pro-Gly-Xaa-Gly)n, wherein n=20-90, and Xaa is a hydrophilic amino acid; and
a hydrophobic block comprising a pentameric amino acid motif (Val-Pro-Gly-Xaa′-Gly)n, wherein n=20-90, and Xaa′ is a hydrophobic amino acid.

2. The biocompatible pharmaceutical composition of claim 1 , wherein the ELP is assembled into a spherical nanoparticle comprising a core into which the Hb, the Hb subunit(s), the Hb fragment(s), or the Hb derivative(s) is encapsulated.

3. The biocompatible pharmaceutical composition of claim 1 , wherein the ELP comprises a pentameric amino acid motif (Val-Pro-Gly-Xaa-Gly)n, wherein n=20-90, and Xaa is Ser or Ile, or a conservative amino acid substitute thereof.

4. The biocompatible pharmaceutical composition of claim 3 , wherein the conservative amino acid substitute of Ser is Thr.

5. The biocompatible pharmaceutical composition of claim 3 , wherein the conservative amino acid substitute of Ile is Leu, Met, or Val.

6. The biocompatible pharmaceutical composition of claim 1 , wherein for the hydrophilic block, the Xaa is selected from the group consisting of Lys, Arg, Asp, Glu, Ser, Thr, Asn, Gln, and His; and for the hydrophobic block, Xaa′ is selected from the group consisting of Ala, Val, Leu, Ile, Pro, Phe, Trp, and Met.

7. The biocompatible pharmaceutical composition of claim 6 , wherein for the hydrophilic block the Xaa is Ser or a conservative amino acid substitute thereof;
and for the hydrophobic block the Xaa′ is Ile or a conservative amino acid substitute thereof.

8. The biocompatible pharmaceutical composition of claim 7 , wherein the conservative amino acid substitute of Ser is Thr; and the conservative amino acid substitute of Ile is Leu, Met or Val.

9. The biocompatible pharmaceutical composition of claim 1 , wherein n=48 for hydrophobic block and n=48 for hydrophilic block.

10. The biocompatible pharmaceutical composition of claim 1 , wherein the Hb, the Hb fragment(s), or the Hb derivative(s) is operatively linked to the C-terminus of the ELP.

11. The biocompatible pharmaceutical composition of claim 1 , wherein the Hb, the Hb subunit(s), the Hb fragment(s), or the Hb derivative(s) is operatively linked to the hydrophobic block of the ELP.

12. The biocompatible pharmaceutical composition of claim 1 , wherein the ELP comprises the amino acid sequence SEQ ID NO: 7.

13. The biocompatible pharmaceutical composition of claim 1 , wherein the Hb, the Hb subunit(s), the Hb fragment(s), or the Hb derivative(s) comprises one or more pharmaceutically acceptable salts thereof.

14. A method of treating a condition due to blood loss and improving subject survival, the method comprising: administering a therapeutic amount of the biocompatible pharmaceutical composition of claim 1 to the subject in need thereof.

Assignees

• Univ Southern California

Inventors

• Despanie Jordan Tremaine
• Guo Hao
• Mackay John Andrew

Classifications

• C07K2319/00

  Fusion polypeptide · CPC title

• A61K38/42

  Haemoglobins; Myoglobins · CPC title

• C07K14/78

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] · CPC title

• C07K14/805 Primary

  Haemoglobins; Myoglobins · CPC title