WO1993001806A1

WO1993001806A1 - Encapsulated perfluorocarbons

Info

Publication number: WO1993001806A1
Application number: PCT/US1992/006132
Authority: WO
Inventors: Tully J. Speaker
Original assignee: Temple University Of The Commonwealth System Of Higher Education
Priority date: 1991-07-23
Filing date: 1992-07-23
Publication date: 1993-02-04

Abstract

An artificial blood substitute comprising Lewis acid - Lewis base salt film microcapsule having a perfluorocarbons encapsulated therein. The Lewis base may be a polyoxyalkylene adduct of an amine, having structure (I) in which, independently, R1=(II), a=7-26, R2=-CH2-O-R1 or CH2-N-(R1)2, R3=(III), b=7-26, and R4=H, CH3 or alkyl, and said Lewis acid may be a polyoxyalkylene derivative of a polymeric acid having structure (IV), in which c- and d-type repeat units are randomly distributed within the polymer, c=70 to 3150, c-type repeat units comprise from 50-90 % of the total polymer, d=70-1750, the sum of c+d is in the range from about 140 to about 3500 and in which R=(V), where R' and R'' independently = H or CH3 and e=7-75, f=7-75, and the sum of e+f=14 to 85.

Description

ENCAPSULATED PERFLUOROCARBONS

Field of the Invention

This invention relates to a microcapsule composition for the delivery of an oxygen and carbon dioxide carrying compound through the vascular system. In particular, the microencapsulated composition is useful as an artificial blood substitute.

Background

A specific type of microcapsulant material and a method of making such material is disclosed in U.S. Patent No. 3,959,457 (of common assignment and partial common inventorship herewith) . This material is comprised of the reaction product produced at the interface boundary of a finely dispersed emulsion, comprising a water immiscible solution of a Lewis base and an aqueous solution of a partially hydrophilic, partially lipophilic Lewis acid.

An improvement in the invention of the '457 patent is disclosed in a presently pending U.S. application (also of common assignment herewith and partial common inventorship) , at least one foreign counterpart of the parent of which has been published prior to the conception of the present invention (see European Patent Application, Publication Number 0 299 205, Publication Date, January 18, 1989) . According to specific aspects of this prior improvement, polyoxethylene-polyoxypropylene or block copolymers thereof comprise a core-forming adjuvant and/or ethylene oxide adducts of ethylenediamine ("tetronomers") comprise a wall-forming adjuvant for use in systems of the '457 patent type. The resultant microcapsules are more robust and more stable physically than the non-adjuvant microcapsules disclosed in the "457 patent.

Perfluorocarbons are known to be useful as blood substitutes but only in the form of dispersions or emulsions. One such known prior art emulsion of perfluorocarbons is marketed under the trade name Fluosol DA. It is understood that this and similar dispersed emulsions exhibit low stability and must be stored at -40°C. Microspheres containing cross-linked hemoglobin and encapsulated hemoglobin have also been prepared for use as a blood substitute. (See U.S. patents 3,875,510 - Kitajima et al., 4,133,874 - Miller et al. and 4,376,059 - Davis et al. )

Encapsulated fluorocarbons have previously been disclosed as tracers for use in the petroleum and petrochemical fields, but the known encapsulants (see U.S. Patents 4,520,109 - Sim onds et al. and 3,964,294 - Shair et al.) are quite different from those used in the present invention.

Heretofore, such perfluorocarbons have been considered poor candidates for encapsulation in systems of the '457 type because their polar character would be expected to destabilize the capsule wall structure.

The extremely low polarity of perfluorocarbons causes them to be incapable of solvating the regions of high charge density associated with ionic centers in systems of the '457 type or of forming hydrogen bonds with water solvating those ionic centers. As a practical result, it has heretofore been impossible to capture perfluorocarbons in charged film microcapsules of the type produced by '457. A technique is described in '457, currently pending U.S. application 07/417,590 and European Patent Application 0 299 205, by which nonpolar materials may be utilized as the dispersed phase and encapsulated. In this technique a solvent of intermediate polarity, such as chloroform, is added to a core material of low polarity, such as light liquid petrolatum, to increase polarity of the dispersed phase. This technique is not applicable to perfluorocarbons because they exhibit such low polarity that they are immiscible with solvents such as chloroform or the even less polar material light liquid petrolatum.

Three other references identified as of possible interest in a patentability search on the present invention are U.S. Patents 4,107,288 - Oppenheim et al., 4,512,762 - Spears and 4,812,445 - Eden et al. The first and third of these pertain to encapsulation techniques different from that used in the present invention and the second, at column 2, lines 37-42, makes reference (though negative) to blood exchange with perfluorocarbons chemicals. Brief Description of the Invention

This invention comprises a microencapsulated perfluorocarbons composition. The invention makes use of a Lewis acid - Lewis base salt microparticulate material similar to that of U. S. Patent No. 3,959,457, with a polyoxyalkylene adduct of an amine (POAAM) , (for example, a polyoxyalkylene adduct of a mono-, di- or poly-amine) as the Lewis base reactant or through use of a polyoxyalkylene derivative of a polymeric acid (PODPA) , for example, a polyoxyalkylene partial ester of a polymeric acid, as the Lewis acid reactant. More specifically, when said Lewis base is a polyoxyalkylene adduct of an amine, said Lewis acid is a polyacid; when said Lewis acid is a polyoxyalkylene derivative of a polymeric acid, said Lewis base is a simple amine.

In a preferred embodiment, the Lewis base polyoxyalkylene adduct of an amine is a tetrapolyoxyethylene adduct of ethylenediamine, also known as a tetronomer. In another preferred embodiment, the Lewis acid polyoxyalkylene derivative of a polymeric acid is the polyethylene glycol ester of polyacrylic acid in which nominally 30 percent of the carboxyl groups of polyacrylic acid are esterified with the glycol.

Most importantly, a perfluorocarbons (examples of which include perfluorotetralin, perfluorobutylamine and perfluorobutyl ether) is disposed in the core of the microcapsule.

It is believed that the pendant polyethers of the POAAM or of the PODPA provide a layer of moderate polarity interposed between the ionic microcapsule wall and the encapsulated perfluorocarbon of low polarity, thus protecting the ionic wall from destabilization by the perfluorocarbons. Preferably, the microcapsules are of about the same size as or somewhat smaller than red blood cells (i.e. about 3 microns in diameter) so that they can circulate in the vascular system as supplements or replacements for red blood cells, either in a blood substitute or in natural blood. For such purposes, they would ordinarily be combined, in a sterile suspension, with suitable additives, such as physiologic electrolytes and macromolecules contributing to the ionic and osmotic balance required of a blood substitute.

Preferably also, the microcapsules of the invention can be associated with a compound or compounds capable of conferring stoichiometric equilibrium to the microcapsule when the microcapsule is administered to an animal. Preferably, the compound is a protein, and more preferably, the compound is albumin. Association of the microcapsule with such a compound may render it more compatible with the blood of the animal into which it is introduced, thereby reducing or eliminating potential adverse side-effects to the animal. This may be especially important when large volumes of suspended microcapsules are administered to the animal.

Detailed Description of the Invention ^'

Generally, Lewis acid - Lewis base salt film microcapsules, of the type to which this invention is directed, are made as follows:

A solution of a suitable Lewis base in a perfluorocarbons is dispersed in an aqueous solution of a Lewis acid to produce a momentary, finely divided emulsion of perfluorocarbon droplets in a continuous aqueous phase. The perfluorocarbons may be, for example, perfluoroalkanes or perfluoroalkenes such as perfluorohexane, perfluoromethyleyclohexane, perfluorodimethylcyclohexane, perfluorotetralin, perfluoromethyltetralin, perfluoromethyldecalin, or perfluoro-5-decene, perfluoroamines such as perfluorotributylamine, perfluoroethers such as perfluorobutyl ether, or halogen substituted perfluorocarbons such as perfluorooctyl bromide.

The Lewis base reactant is selected from a simple amine capable of reacting with a polyoxyalkylene derivative of a polymeric acid and a polyoxyalkylene adduct of a mono-, di-, or poly-amine (POAAM). The POAAM has the following generic structure:

CH₂-R₂ / R-_^-N

\

CH--R3

in which, independently,

R-_j_ = H(-0-CH₂-CH)a, a=7-26 R₂ = -CH2-0-R_! or CH₂-N-(R_!)₂

R-.

I R₃ = R₂ or - (CH₂-N-CH₂)_b-R₂, b=7-26

and

R4 = H, CH3 or alkyl. The Lewis acid reactant is selected from a polyacid capable of reacting with a polyoxyalkylene adduct of an amine and a polyoxyalkylene derivative of a polymeric acid (PODPA) . The PODPA has the following generic structure:

H(C-C-)_c(C-C)_dH C IOOH CIOOR,

C=70-3150 and d=70-1750

in which c- and d- type repeat units are randomly distributed within the polymer, c=70 to 3150, c-type repeat units comprise from 50-90% of the total polymer, d=70-1750, the sum of c + d is in the range from about 140 to about 3500

and in which:

R = - (CH- -O-) - ( H-CH-O)_fH

where

R'and R" independently = H or CH3 and e=7 to 75, f=7-75, and the sum of e + f = 14 to 85. In the preferred embodiment, c type repeat units comprise approximately 70% of the total polymer, R'=H, e=14 and f=0.

For reasons not entirely clear, the general class of POAAM as the Lewis base and the general class of PODPA as the Lewis acid are not well adapted for concurrent use. However, there may be specific compounds having the PODPA and POAAM structures which are capable of producing the microcapsule of the claimed invention. When the Lewis base reactant is a POAAM, the Lewis acid is a polymeric acid capable of reacting with a polyoxyalkylene adduct of an amine, such as acacia, Arabic acid, carboxymethylcellulose, ghatti gum, guar gum, polyacrylic acid and the like. As such, the polyacid is not likely to include the polyoxyalkylene derivative of a polymeric acid (PODPA) . The preferred concentration range for the Lewis acid is 1%-10% w/v.

Alternatively, in the absence of a POAAM Lewis base, the Lewis acid reactant is a PODPA, and the Lewis base is a simple amine capable of reacting with a polyoxyalkylene derivative of a polymeric acid, a sufficiently strongly basic mono-, di- or poly- amine such as n-hexylamine, octadecylamine, ethylenediamine, piperazine, 2,4- imethyIpentylamine, tetraethylenetriamine, polyethylenei ine and the like. As such, the simple amine is not likely to include the POAAM. The preferred concentration range of the Lewis base is restricted by the limiting solubility of the Lewis base in the perfluorocarbons employed as core material. Typically, the concentration of amine is in the range 0.1 to 10%. While the exact concentration of the solution of amine serving as Lewis base is not critical, it is important to use a total amount of base stoichiometrically equivalent to the non-esterified ionizable carboxylate functions of the PODPA.

In accordance with the present invention, the Lewis base is dissolved in the perfluorocarbons (PFC) resulting in a PFC-Lewis Base (PFC-LB) phase. The Lewis acid is dissolved in the aqueous solution, prior to combining the aqueous solution with the PFC-LB phase. A momentary, finely dispersed emulsion of PFC droplets in a continuous aqueous phase is produced with rapid stirring. The Lewis acid and Lewis base must be substantially soluble in the appropriate phase. Accordingly, in one of the preferred embodiments, the Lewis base, a POAAM, is dissolved in the PFC phase, and Arabic acid is dissolved in the aqueous phase. In the other preferred embodiment, the Lewis base 2,4-dimethylpentylamine is dissolved in the PFC phase and the Lewis acid, a PODPA, is dissolved in the aqueous phase.

In the present invention, perfluorocarbons encapsulation efficiencies near 60% are easily achieved with a 1:1 ratio of aqueous (Lewis acid-containing) phase to non-aqueous (Lewis base-PFC-containing) phase, essentially 100% of the perfluorocarbons may be encapsulated if the initial nonaqueous to aqueous phase ratio is adjusted to 2:5.

General Procedures for Forming Microcapsules

In all instances where an aqueous solution is utilized as the continuous phase for the dispersion of the PFC-Lewis base, it is preferred, but not essential, that the PFC-Lewis base be slowly and steadily added to the aqueous solution over a period of approximately 30 seconds. Generally, solutions are prepared and reactions take place at room temperature, unless otherwise stated. Any of several means to disperse the PFC-Lewis base in the aqueous medium may be employed including:

a. vigorously stirring the solution with a magnetically driven stirring bar at a nominal shear rate, generally 700 or more cm/s;

b. vigorously mixing the solution with a multi-orifice axial turbine (such as a Brinkmann homogenizer PT10/35 and generator PST/10, Brinkmann Instruments, Westbury, N.Y.) at a nominal setting of 5; or c. vigorously agitating the solutions with an ultrasonic probe (such a Heat Systems model W185D, Ultrasonics, Inc., Plainview, N.Y.) at a nominal output of 100 watts.

General Method of Preparing A Microencapsulated Perfluorocarbons

A 10 ml volume of a 10% w/v aqueous solution of Arabic acid was mixed with a stoichiometrically equivalent solution of a tetronomer, (i.e., ca 2.2g), such as Tetronic 702, a product of BASF-Wyandotte Corp., Wyandotte, IL, in 5 ml of perfluorotetralin. The mixture was insonated at 20 kilohertz for 1-5 minutes at a power output of about 100 watts. The sonic energy dispersed the perfluorocarbons phase into small droplets which had the momentary characteristic of an emulsion stabilized in part by the surfactant properties of Arabic acid. Studies with small volume (2-5 ml) formulations show that longer msonation produces finer dispersions but does not appear to markedly increase encapsulation efficiency.

The reaction mixture was centrifuged (10,000 gravity minutes) and the supernatant removed. Distilled water was added and the pellet of microcapsules was resuspended by vortexing. The suspension was allowed to stand a few minutes to permit unencapsulated perfluorocarbons phase to separate, and the microcapsular suspension was harvested. The microcapsules were resuspended in distilled water and centrifuged as needed to remove unreacted manufacturing components. The washed product was rendered as a flowable concentrate by a final centrifugation. The resulting preparation of microencapsulated perfluorocarbons is stable at 20°C for about one month. Similar experiments were conducted to compare the encapsulation of perfluorocarbons with different Lewis acids. As set forth in Table I below, the control 1, polyethylene glycol alone is not capable of encapsulating perfluorocarbons nor does control 2, polyacrylic acid alone, or control 3, the simple non- esterified combination of polyethylene glycol mixed with polyacrylic acid encapsulate perfluorocarbons, in an attempted wall forming reaction using 2,4- dimethylpentamine as Lewis base.

In contrast, experiment 4, using polyacrylic acid (ca 30%) esterified with polyethylene glycol as the Lewis acid and the same 2,4-dimethylpentamine as Lewis base does effectively 100% encapsulate perfluorocarbons.

- 12 -

Table I

_Controls Experiment

1 2 4

where

PEG = polyethylene glycol 600 ⁽ave MW = 600⁾

PAA = polyacrylic acid (ave MW = 250000, mEqWT = 72 mg⁾

PODPA = polyethylene glycol 600 polyacrylate

₍degree of es_terification: ca 30%, mEqWt == 350 mg^} AMI_NE = 2,₄-dimethylpentylamine (MW = 115, mEqWt = ¹¹5 mg) AQ = distilled water PFC = perfluorocarbons (perfluorodec-5-ene⁾

To assess the effect of perfluorocarbons microcapsules m vivo in a mammal, the following experiment was performed. Mice were administered suspensions of perfluorocarbons microcapsules (or an equal volume of sterile saline as a control) intravenously into the tail vein. The microcapsules contained the polyethylene glycol poly-acrylate macromolecule and were prepared as described above. Each mouse received 0.05 mL of suspension which is approximately 5 to 7% of the total blood volume of the animals. The animals were sacrificed in groups of five at logarithmically incrementing times of 3, 10, 30, 100, 300, 1000 and 3000 minutes. Importantly, all of the test and control mice lived until sacrificed and none displayed any indication of toxicity except for transient quiescence. Thus, perfluorocarbons encapsulation, as described above, is tolerated by these animals without undue or remarkable distress.

Additives such as physiologic electrolytes and macromolecules may be incorporated in the microencapsulated perfluorocarbons suspensions. These additives contribute to and adjust for the ionic and osmotic balance required of a blood substitute, whose serum specifications are well known, or a perfusion fluid.

Additives may be especially important when encapsulated perfluorocarbons are used as a means to replace larger volume fractions of blood. Blood not only carries oxygen, but also contains nutrients, ions and colloids, the most abundant of which is albumin. Such compounds, when dissolved in blood plasma, contribute to the osmotic tension of blood and are vital to maintaining fluid balance in all perfused tissues. It is well known in the art that perfluorocarbons emulsions exert a "protein stripping" action on blood. The general phenomenon of aggregration of proteins with perfluorocarbons is described in Biro et al. (CRC Critical Reviews in Oncology Hematology, Vol. 6, No. 4, pp 311-374, "perfluorocarbons Blood Substitutes") . "Protein stripping" is understood to be the association of protein which no-πtially exists in solution in plasma with microparticulate material to form a bound complex comprising the microparticle (emulsion droplet) , the adherent protein, and possibly other macromolecules. Thus, replacement of large volume fractions of blood with encapsulated perfluorocarbons may result in dilution of ions and colloids, in "protein stripping", and in subsequent hypo-osmolarity of blood.

To circumvent this potential problem, the microencapsulated perfluorocarbons suspension can be combined with, or be treated with, a compound or compounds which when associated with microcapsules, may reduce or eliminate depletion of blood components, in particular albumin, which are essential for maintaining normal osmolarity of blood. Larger volumes of microcapsules when so treated, can then be administered to an animal without inducing potentially adverse side- effects. Elimination or reduction of potentially adverse effects may be accomplished by simply administering microcapsules to animals in conjunction with a sufficient quantity of a compound, without prior treatment of the microcapsule with that compound. Alternatively, the microcapsule of the invention may be pretreated with such a compound. An example of such pretreatment is now described below.

To explore whether treatment of microcapsules with such a compound could be tolerated in mice, a microcapsule suspension prepared as described above, was treated with albumin prior to administration of the suspension to mice. A solution of mouse albumin (Sigma Chemical Co.) at a concentration of 20 mg in 2 mL of sterile 0.9% saline was prepared. This solution was added to an equal volume of encapsulated perfluorocarbons suspension and the mixture was incubated at 38°C for 30 minutes. Albumin treated microcapsules were separated from unreacted albumin by centrifugation and resuspended in 2 mL of sterile saline. Five mice were inoculated with 0.2 L aliquots of treated perfluorocarbons microcapsules into the tail vein. This volume represents approximately 20-28% of the total blood volume of a mouse. All five animals remained viable for more than 300 minutes following inoculation. One animal died within one day and another died within two days following inoculation. The remaining animals survived and appeared healthy for a least one week. Because the sterility of the suspension could not be guaranteed and because the large volume of the injection represents a substantial fluid stress to the animals, the death of the two animals was not surprising. Notwithstanding, the fact that three of the animals survived and remained healthy for greater than one week is evidence that pretreatment of microcapsules with albumin can be tolerated by the animals. Furthermore, these data demonstrate that this treatment has potential use when circumstances necessitate that large volumes of microcapsules be administered to an animal. It is well known in the art that animals such as mice represent excellent models for therapies and treatments of other animals and humans. Because mice which had received either albumin-treated or untreated perfluorocarbons microcapsules not only survived but also appeared to be healthy, such microcapsules may also be tolerated and provide useful therapies to other animals and to humans. It is not necessary to limit pretreatment of microcapsules to albumin alone. Conceivably, perfluorocarbons microcapsules can be pretreated with any protein, colloids, or even inorganic ion components of blood such that they are adjusted to stoichiometric equilibrium with that of the blood of the animal or human to whom they will be administered. One of ordinary skill in the art, without undue experimentation will know following the general methods described above, how to pretreat microcapsules with other compounds such as non- blood proteins, other blood proteins, and colloids and ions associated with blood. Similarly, when microcapsules are to be administered to an animal in - conjunction with a compound, the invention need not be limited to albumin alone. Other compounds such as those described above, are also suitable for administration in conjunction with microcapsules.

When albumin is the blood component of choice, the type of albumin used should be compatible with the species of animal being treated. Many species specific preparations of albumin, including human albumin, are available commercially. In addition, when albumin is administered to the animal in conjunction with the microcapsule of the invention, preferably the suspension of microcapsules and albumin should contain up to 5% weight by volume of albumin, which is the approximate maximal concentration of albumin in blood. Higher concentrations of albumin may be used if medical conditions so indicate.

Generally, the microencapsulated perfluorocarbons of the present invention is stable at room temperature. It should be capable of circulating freely in the vascular system; taking up, carrying and releasing dissolved oxygen to tissues; and capturing carbon dioxide from tissues and releases it in the lungs.

It is believed that the material of the present invention will be useful as (or in) perfusion fluids in hospital surgical suites, and as (or in) a temporary substitute or replacement for human blood, for example, when supplies of real human blood are limited or unavailable, when inadequate time or facility is available to determine the blood type of the intended recipient, such as at sites of accidents, disasters, catastrophe scenes, battlefields and hospital emergency rooms; when there is concern for transmission of blood- borne diseases, such as AIDS and hepatitis, from donor to recipient; when the use of human blood is rejected for religious or ethical reasons. The microencapsulated perfluorocarbons of this invention may also benefit athletes, marathon runners for example, anemics, and animals, such as race horses, by providing a means to improve the oxygen and carbon dioxide exchange capacity of the vascular system of such subjects.

Problems with other known perfluorocarbons used as artificial blood are overcome by the present invention, including stability problems, as the microencapsulated perfluorocarbons of the present invention are stable, and do not coalesce, at 20°C for about one month. The microencapsulated perfluorocarbons of the present invention eliminates the need for thawing the blood substitute and oxygenating the perfluorocarbons prior to administration. Furthermore, the microencapsulated perfluorocarbons of the invention eliminates the need for blood typing prior to use. Accordingly, the microencapsulated perfluorocarbons of the claimed invention may be used more rapidly than conventional blood substitutes. In addition to use as a blood substitute or a perfusion fluid, the microencapsulated perfluorocarbons of the present invention also has utility in other applications in which stable compositions including perfluorocarbons have been shown to be useful, such as a contrast agent in medical imaging. Perfluorocarbon- containing perfusion fluid may also be used during surgery and for wound healing to increase oxygen carrying capacity to damaged and surrounding tissues.

While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and equivalent variations of this invention may be devised by those skilled in the art without departing from the true spirit and scope of this invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

What is Claimed:

1. A Lewis acid-Lewis base salt film microcapsule having a microcapsule wall and perfluoro¬ carbon as a core material, said Lewis acid is selected from a polyacid capable of reacting with a polyoxyalkylene adduct of an amine and a polyoxyalkylene derivative of a polymeric acid having the structure

H(C-C-)_c(C-C)_dH C IOOH CIOOR

in which c- and d- type repeat units are randomly distributed within the polymer, c=70 to 3150, c-type repeat units comprise from 50-90% of the total polymer, d=70-1750, the sum of c + d is in the range from about 140 to about 3500 and in which:

R = - ( H- -O-) - (CH- -O)_fH

where

R'and R" independently = H or CH3 and e=7-75, f=7-75, and the sum of e + f = 14 to 85,

and said Lewis base is selected from a simple amine capable of reacting with a polyoxyalkylene derivative of a polymeric acid and a polyoxyalkylene adduct of an amine having the structure CH₂-R₂ / R-.-N \ CH2-R3 in which, independently,

R₄

I R-_ = H(-0-CH₂-CH)_a, a=7-26

R₂ = -CH2-0-R_! or CH -N-(R₁)

Ri I

R₃ = R₂ or - (CH₂-N-CH₂)b-R₂, b=7-26 and

R₄ = H, CH3 or alkyl,

wherein when said Lewis base is a polyoxyalkylene adduct of an amine said Lewis acid is a polyacid, and when said Lewis acid is polyoxyalkylene derivative of a polymeric acid said Lewis base is a simple amine.

2. A microcapsule of claim 1 wherein said per¬ fluorocarbon is selected from the group consisting of perfluoroalkanes, perfluorocycloalkanes, perfluoroalkenes, perfluoroamines, perfluoroethers, or halogen substituted perfluorocarbons.

3. A microcapsule of claim 2 wherein said perfluorocarbons is selected from the group consisting of perfluorotetralin, perfluoromethyldecalin, perfluorohexane, perfluoromethylcyclohexane, and perfluoro-1,3-dime hylcyclohexane.

4. A microcapsule of claim 2 wherein said perfluorocarbons is perfluorotributylamine.

5. A microcapsule of claim 2 wherein said perfluorocarbons is perfluorobutyl ether.

6. A microcapsule of claim 2 wherein said perfluorocarbons is perfluorooctyl bromide.

7. A microcapsule of claim 1 wherein said polyacid is selected from the group consisting of Arabic acid, acacia gum, carboxymethylcellulose, ghatti gum, guar gum, and polyacrylic acid.

8. A microcapsule of claim 1 wherein said simple amine is selected from the group consisting of n-hexylamine, 2,4-dimethylpentylamine, octadecylamine, ethylenediamine, piperazine, tetraethylenetriamine and polyethyleneimine.

9. A microcapsule of claim 1 wherein said polyacid is Arabic acid.

10. A microcapsule of claim 9 wherein said polyoxyalkylene adduct of an amine is a polyoxyethylene adduct of ethylenediamine.

11. A microcapsule of claim 1 wherein said polyoxyalkylene adduct of an amine has polyoxyalkylene chains disposed between said microcapsule wall and said core material.

12. In a blood substitute, comprising the improvement as an artificial red blood cell, a Lewis acid-Lewis base salt film microcapsule having a microcapsule wall and perfluorocarbons as a core material, wherein said Lewis acid is selected from a polyacid capable of reacting with a polyoxyalkylene adduct of an amine and a polyoxyalkylene derivative of a polymeric acid having the structure

H(C-C-)_c(C-C)_dH C IOOH CIOOR

R = - (CH-CH-O-)p- ( H- -O)fH

where

R'and R" independently = H or CH₃ and e=7-75, f=7-75, and the sum of e + f = 14 to 85,

and said Lewis base is selected from a simple amine capable of reacting with a polyoxyalkylene derivative of a polymeric acid and a polyoxyalkylene adduct of an amine having the structure

CH₂-R₂

/

Rl-N

\ CH₂-R₃ in which, independently,

R₄

I

R₁ = H(-0-CH₂-CH)_a, a=7-26 R₂ -s -CH2-O-R-. or CH₂-N-(R₁)₂

*ι

I

R₃ = R₂ or - (CH₂-N-CH₂)b-R₂, b=7-26 and

R₄ = H, CH3 or alkyl,

13. A blood substitute of claim 12 wherein said polyacid is selected from the group consisting of acacia, Arabic acid, carboxymethylcellulose, ghatti gum, guar gum, and polyacrylic acid.

14. A microcapsule of claim 12 wherein said simple amine is selected from the group consisting of n-hexylamine, 2,4-dimethylpentylamine, octadecylamine, ethylenediamine, piperazine, tetraethylenetriamine and polyethyleneimine.

15. In a perfusion fluid, comprising the improvement as an artificial red blood cell, a Lewis acid -Lewis base salt film microcapsule having a microcapsule wall and perfluorocarbons as a core material, wherein said Lewis acid is selected from a polyacid capable of reacting with a polyoxyalkylene adduct of an amine and a polyoxyalkylene derivative of a polymeric acid having the structure

H(C-C-)_c(C-C)_dH

C IOOH CIOOR

in which c- and d- type repeat units are randomly distributed within the polymer, c=70 to 3150, c-type repeat units comprise from 50-90% of the total polymer, d=70 to 1750, the sum of c + d is in the range from about 140 to about 3500 and in which:

R = - (CH-CH-O-)p- (CH-CH-O)_fH

where

CH₂-R₂

/

Rl-N

\

CH₂-R₃ in which, independently,

R₄

I

R-_ = H(-0-CH₂-CH)_a, a=7-26 R₂ = -C^-0-R_! or CH₂-N-(R_!)₂

I R₃ « R₂ or - (CH₂-N-CH₂)b-R₂, b=7-26 and

R = H, CH₃ or alkyl,

16. A blood substitute of claim 15 wherein said polyacid is selected from the group consisting of acacia, Arabic acid, carboxymethylcellulose, ghatti gum, guar gum, and polyacrylic acid.

17. A blood substitute of claim 15 wherein said simple amine is selected from the group consisting of n-hexylamine, 2,4-dimethylpentylamine, octadecylamine, ethylenediamine, piperazine, tetraethylenetriamine and polyethyleneimine.

18. A method of making a microencapsulated perfluorocarbon having a wall and a core, comprising combining: a. an aqueous phase including a Lewis acid wherein said Lewis acid is selected from a polyacid and a polyoxyalkylene derivative of a polymeric acid with b. a non-aqueous phase including

(1) a stoichiometric equivalent of a Lewis base, wherein said Lewis base is selected from a simple amine and a polyoxyethylene adduct of an amine and

(2) a perfluorocarbons selected from the group consisting of perfluoroalkanes, perfluorocycloalkanes, perfluoroalkenes, perfluoroamines, perfluoroethers, or halogen substituted perfluorocarbons; c. dispersing said aqueous and said non-aqueous phases together to momentarily form an emulsion, wherein said perfluorocarbon is disposed in said core.

19. The microcapsule of claim 1, wherein said microcapsule further comprises a compound capable of conferring stoichiometric equilibrium to said microcapsule when said microcapsule is administered to an animal.

20. A blood substitute comprising the microcapsule of claim 19.

21. A perfusion fluid comprising the microcapsule of claim 19.

22. The method of claim 18, said method further comprising adjusting said microcapsule to stoichiometric equilibrium with mammalian blood, said adjustment comprising mixing said microcapsule with a compound capable of conferring stoichiometric equilibrium to said microcapsule, and then removing unreacted compound from said microcapsule.

23. The microcapsule of claim 19, wherein said compound is a protein.

24. The microcapsule of claim 23, wherein said protein is albumin.

25. A method of treating a mainmal requiring a blood substitute comprising administering to said mammal the microcapsule of claim 1.

26. A method of treating a mammal requiring a blood substitute comprising administering the microcapsule of claim 19.

27. The microcapsule of claim 1 being associated with albumin.

28. The microcapsule of claim 27 being associated with up to 5% weight by volume of albumin.

29. A method of treating a mammal requiring a blood substitute comprising administering to said mammal the microcapsule of claim 27.