Blood substitute composition and method of use

Abstract

The present disclosure provides oxygen-carrying nanoparticles, methods of making the nanoparticles, and methods of using the nanoparticles to carry oxygen in blood.

What is claimed is: 
     
         1 . A nanoparticle having a substantially bi-concaved disc shape, wherein the nanoparticle comprises an aqueous core, a bi-layered shell comprising an amphiphilic polymer, and a payload; wherein the bi-layered shell has a hydrophilic outer layer, a hydrophilic inner layer, and a hydrophobic region between the hydrophilic outer layer and the hydrophilic inner layer; the amphiphilic polymer comprises a branched, amine-containing polymer linked to a lipid; and the payload comprises an oxygen-carrying agent, an allosteric effector, and a reducing agent. 
     
     
         2 . The nanoparticle of  claim 1 , wherein the amphiphilic polymer comprising the hydrophilic outer layer of the shell is derivatized with polyethylene glycol, such that the particle has a zeta potential of about −15 mV to about +15 mV. 
     
     
         3 . The nanoparticle of  claim 1 , wherein the average diameter of the nanoparticle is from about 150 nm to about 300 nm, and the average height of the nanoparticle is from about 30 nm to about 80 nm. 
     
     
         4 . The nanoparticle of  claim 1 , wherein the nanoparticle comprises a through-hole or a depression. 
     
     
         5 . The nanoparticle of  claim 1 , wherein the oxygen-carrying agent is synthetic hemoglobin or naturally occurring hemoglobin. 
     
     
         6 . The nanoparticle of  claim 1 , wherein the allosteric effector is selected from the group consisting of 2,3-diphosphoglycerate (2,3-DPG), inositol hexaphosphate (IHP), pyridoxal-phosphate (PLP), and 2-[4-[[(3,5-dimethylanilino carbonyl]methyl]-phenoxy]-2-methylpropionic acid. 
     
     
         7 . The nanoparticle of  claim 1 , wherein the reducing agent is selected from the group consisting of leucomethylene blue, glutathione and ascorbate. 
     
     
         8 . The nanoparticle of  claim 1 , wherein the nanoparticle comprises about 3000 to about 10,000 hemoglobin molecules. 
     
     
         9 . The nanoparticle of  claim 1 , wherein the nanoparticle comprises about 20% to about 60% (w/v) hemoglobin or about 30% to about 60% (w/v) hemoglobin. 
     
     
         10 . The nanoparticle of  claim 1 , wherein the nanoparticle does not substantially sequester nitric oxide. 
     
     
         11 . The nanoparticle of  claim 1 , wherein the nanoparticle limits the oxidation of hemoglobin to about 10% or less of the total concentration of hemoglobin in the nanoparticle. 
     
     
         12 . The nanoparticle of  claim 1 , wherein the branched, amine-containing polymer is selected from the group consisting of a polyethyleneimine branched polymer, a PAMAM dendrimer, a star polymer, and a graft polymer. 
     
     
         13 . The nanoparticle of  claim 1 , wherein the lipid is a fatty acid. 
     
     
         14 . The nanoparticle of  claim 13 , wherein the fatty acid is selected from the group consisting of 10,12-pentacosadiynoic acid, hexadecyloctadecanoic acid, cholanic acid, linoleic acid, C24-pentacosadiynoic acid, and palmitic acid. 
     
     
         15 . A process for the preparation of a nanoparticle, the nanoparticle having a substantially bi-concaved disc shape, wherein the nanoparticle comprises an aqueous core, a bi-layered shell comprising an amphiphilic polymer, and a payload comprising an oxygen-carrying agent, an allosteric effector, and a reducing agent; the process comprising: (a) hydrophobically modifying a branched polymer by covalently conjugating lipids to at least 25% of the free reactive groups of the branched polymer to form an amphiphilic polymer; (b) mixing the amphiphilic polymer with a non-polar solvent, and agitating the mixture to form a plurality of inverted micelles comprising the amphiphilic polymer; (c) mixing the payload with a polar solvent and adding the payload mixture to the mixture from (b) to form a bi-phasic mixture, and agitating the bi-phasic mixture to transfer the payload to the inverted micelles; and (d) agitating the inverted micelles comprising the payload in the presence of heat and an aqueous solvent to form the bi-concaved disc shaped nanoparticles comprising the payload. 
     
     
         16 . The process of  claim 17 , wherein the payload is contained within the interior of the nanoparticle. 
     
     
         17 . A blood substitute composition, the composition comprising a nanoparticle of  claim 1 . 
     
     
         18 . The composition of  claim 17 , wherein the composition is a solution or a lyophilized powder.