Blood substitutes and uses thereof

Patent metadata

Field	Value
Publication number	US-9387152-B2
Application number	US-201113805245-A
Country	US
Kind code	B2
Filing date	Jun 28, 2011
Priority date	Jun 28, 2010
Publication date	Jul 12, 2016
Grant date	Jul 12, 2016

Abstract

The invention relates to blood substitute compositions, and methods of use thereof. Described herein are compositions in which hemoglobin is maintained substantially in the reduced form of hemoglobin as opposed to the oxidized methemoglobin form through inclusion of an oxido-reductase enzyme and reducing agent within a vesicle with the hemoglobin. The vesicles additionally can comprise a dismutase, a catalase, and an electron acceptor, each of which contribute to either the maintenance of hemoglobin in the active oxygen carrying state or provide a benefit not achieved with free hemoglobin.

First claim

The invention claimed is:

1. A method of encapsulating a blood substitute composition, the method comprising the steps of:
a. mixing a lipid preparation with a composition comprising an oxido-reductase enzyme, a dismutase, a catalase, a reducing agent, and an electron acceptor;
b. extruding the mixture to form lipid vesicles; and
c. isolating lipid vesicles, wherein the lipid preparation comprises at least one lipid selected from the group consisting of: DPPC, DSPC, DOPC, DLPC, DPPG, DSPG, DHPG, DOPG, DSPEG PEG 5000, DSPE-PEG200, and DOPE-PEG5000,
and, wherein electron acceptors include but are not limited to: methylene blue, sodium 2,6-dibromophenol-indophenol, sodium 2,6-dichlorophenol-indophenol, sodium o-cresol indophenols, indigotetrasulfonic acid, indigotrisulfonic acid, indigo carmine, indigomono sulfonic acid, phenosafranin, safranin T, neutral red, and thionine.

2. The method of claim 1 , wherein the oxido-reductase enzyme comprises DT-diaphorase.

3. The method of claim 1 , wherein the dismutase comprises superoxide dismutase.

4. The method of claim 1 , wherein the reducing agent comprises NADH or NADPH.

5. The method of claim 1 , wherein the electron acceptor comprises methylene blue.

6. The method of claim 1 , wherein the lipid preparation comprises:
a. at least one lipid selected from the group consisting of: DPPC, DSPC, DOPC, and DLPC;
b. at least one lipid selected from the group consisting of: DPPG, DSPG, DHPG, and DOPG; and
c. at least one lipid selected from the group consisting of: DSPEG PEG 5000, DSPE-PEG200, and DOPE-PEG5000.

7. The method of claim 1 , wherein the size of the isolated vesicles is comprised between 150 nm to 500 nm inclusive.

8. The method of claim 1 , wherein the size of the isolated vesicles is comprised between of 180 nm to 220 nm inclusive.

9. The method of claim 1 , wherein the size of the isolated vesicles is 200 nm.

10. The method of claim 1 , wherein the lipid preparation comprises: Cholesterol, DPPC, DSPE-PEG5000, and DPPG.

11. The method of claim 10 , wherein the ratio of Cholesterol, DPPC, DSPE-PEG5000, and DPPG is respectively 5:5:0.0333:1.

Assignees

• Yarmush Martin L
• Yang Eric
• Konry Tali
• Seker Erkin
• Sarin Dhruv
• Rai Prakash
• Gen Hospital Corp

Inventors

• Yarmush Martin L
• Yang Eric
• Konry Tali
• Seker Erkin
• Sarin Dhruv
• Rai Prakash

Classifications

• A61K9/0026 Primary

  Blood substitute; Oxygen transporting formulations; Plasma extender · CPC title

• A61J3/07 Primary

  into the form of capsules or similar small containers for oral use · CPC title

• A61K9/1271

  Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• C12Y115/01001

  Superoxide dismutase (1.15.1.1) · CPC title