Scaled-up production of liposome-encapsulated hemoglobin

• US 4,776,991 A
• Filed: 08/29/1986
• Issued: 10/11/1988
• Est. Priority Date: 08/29/1986
• Status: Expired due to Fees

• Current Assignee: the united states of america as represented by the secretary of the navy
• Original Assignee: the united states of america as represented by the secretary of the navy
• Inventors: Beissinger, Richard L., Farmer, Martha C.
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US06/901,710
• Time in Patent Office: 774 Days
• Field of Search: 264/4.3, 264/4.6, 264/4.1, 428/402.2, 424/450, 436/829, 514/6, 514/832, 514/833
• US Class Current: 264/4.3
• CPC Class Codes: A61K 38/42 Haemoglobins; Myoglobins
• CPC Class Codes: A61K 9/1277 Processes for preparing; Proliposomes
• CPC Class Codes: A61P 7/08 Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock artificial tears A61P27/04
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

A method for making sterile liposome-encapsulated hemoglobin having an oxn carrying capacity of 20 vol/%, a half life of 15-20 hours as measured in mice, with the liposome made from chloroform, HSPC, cholesterol, negatively charged DMPG and alpha-tocopherol.

Claims

1. A process for producing at least 10 mls/min of hemoglobin encapsulated in a liposome wherein said encapsulated hemoglobin has an in vivo half-life in mice of at least 15 hours comprising the steps of:

forming a liposome from a combination consisting essentially of;

(a) a compound selected from the group consisting of hydrogenated soy phosphatidylcholine and distearoyl phosphatidylcholine,

(b) Cholesterol,

(c) Dimyristoyl phosphatidyl gylcerol, and

(d) alpha-tocopherol,

combining said liposome with a sterile dispersion of stoma-free hemoglobin to form a hemoglobin/liposome mixture;

encapsulating the hemoglogin in said liposome by pressurizing said mixture in a first chamber, projecting said mixture into a second chamber having a lower pressure than said first chamber through at least two orifices forming streams which impinge against one another, said impinging streams causing cavitation and agitation in the mixture contained in said second chamber, recirculating said mixture until the average particle size is about 0.20 microns, and

filtering said encapsulated hemoglobin mixture to remove impurities including unencapsulated hemoglobin to produce a sterilized product.

2. The process of claim 1 wherein said orifices are disposed at 180°

to one another so that said fluid streams impinge directly against one another.

3. The process of claim 2 wherein said orifices are rectangularly defined micro-channels.

4. The process of claim 1 wherein said mixture is pressurized in said first chamber to a pressure between 2000 and 5000 psi.

5. The process of claim 1 wherein liposome combination is in the molar ratio of 5:

4;

1;

0.2 respectively.