Patent EP0195558A2 successfully refreshed from Google Patents.
EP0195558A2
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EP
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Composition of alpha-alpha- cross-linked hemoglobins
Assignee: University of Iowa Research Foundation
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Filing: 1986-03-06
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Granted: —
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Expires: 2006-03-06
Abstract
Abstract A high yield method of production of a cross-linked hemoglobin derivative suitable for use as a blood substitute and plasma expander which is cross-linked between Lys 99 Alpha, and Lys 99 Alpha 2 in high percentage by adding the cross-linker in the presence of an added polyanion which blocks competing reactions at other sites of the protein.
Inventors
- 👤 Joseph Alan Walder
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 09:39
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/795; C07K14/805; A; A61; A61P; A61P7/00; A61P7/08; A61K; A61K38/00
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
62 words
A pharmaceutical composition for use as a blood substitute and blood plasma expander characterized in that it comprises a therapeutically effective amount of Lys 99 Alpha 1 to Lys 99 Alpha 2 intra-molecularly cross-linked, stroma-free hemoglobin substantially free of hemoglobin modified at other sites, soluble in aqueous and physiological fluids and capable of reversibly binding oxygen and a pharmaceutically acceptable carrier. 1
Claim 2
Dependent Claim
62 words
A pharmaceutical composition for use as a blood substitute and blood plasma expander characterized in that it comprises a therapeutically effective amount of Lys 99 Alpha 1 to Lys 99 Alpha 2 intra-molecularly cross-linked, stroma-free hemoglobin substantially free of hemoglobin modified at other sites, soluble in aqueous and physiological fluids and capable of reversibly binding oxygen and a pharmaceutically acceptable carrier. 1
Claim 3
Dependent Claim
19 words
A composition according to claim 1, characterized in that the alpha-alpha cross-link is with an amino group-specific cross-linking agent.
Claim 4
Dependent Claim
16 words
A composition according to claim 2, characterized in that the cross-linking agent is an acylating agent.
Claim 5
Dependent Claim
17 words
A composition according to claim 3, characterized in that the cross-linking agent is a diester cross-linking agent.
Claim 6
Dependent Claim
18 words
A composition according to claim 4, characterized in that the cross-linking agent is a phenyl ester cross-linking agent.
Claim 7
Dependent Claim
15 words
A composition according to claim 5, characterized in that the cross-linking agent is bis(5,5-dibromasalicyl) fumerate.
Claim 8
Dependent Claim
25 words
A composition according to claim 1, characterized in that the carrier is liquid and the composition contains from about 1% to about 10% of hemoglobin
Claim 9
Dependent Claim
28 words
A composition according to claim 1, characterized in that the hemoglobin is also modified with a second reagent introducing a negatively charged group at the 2,3-diphosphoglycerate binding site.
Claim 10
Dependent Claim
14 words
A composition according to claim 8, characterized in that the reagent is mono(3,5-dibromosalicyl) fumerate.
Claim 11
Independent Claim
74 words
A method of replacing or increasing the circulating blood volume or increasing oxygen delivery to tissues in man or animal species characterized in that it comprises transfusing into the blood circulatory system a blood volume expander which consists essentially of Lys 99 Alpha to Lys 99 Alpha 2 intramolecularly cross-linked, stroma-free hemoglobin which is substantially free of hemoglobin derivatives modified at the sites, having a molecular weight of about 64,000 in an isotonic solution.
Claim 12
Independent Claim
74 words
A method of replacing or increasing the circulating blood volume or increasing oxygen delivery to tissues in man or animal species characterized in that it comprises transfusing into the blood circulatory system a blood volume expander which consists essentially of Lys 99 Alpha to Lys 99 Alpha 2 intramolecularly cross-linked, stroma-free hemoglobin which is substantially free of hemoglobin derivatives modified at the sites, having a molecular weight of about 64,000 in an isotonic solution.
Claim 13
Dependent Claim
24 words
A method according to claim 10, characterized in that the hemoglobin is also modified with a negatively charged group at the 2,3-diphosphoglycerate binding site.
Claim 14
Independent Claim
82 words
A method of producing in commercially significant yields, a modified hemoglobin which is capable of functioning as a blood substitute and blood plasma expander characterized in that it comprises isolated unmodified hemoglobin or preparing a crude red cell lysate, deoxygenating the hemoglobin cross-linking the deoxygenated hemoglobin intramolecularly between Lys 99 Alpha 1 and Lys 99 Alpha 2 in the presence of an added polyanion selected from the group consisting of 2,3-diphosphoglycerate, inositol hexaphosphate, or inositol hexasulfate and purifying the alpha-alpha cross-linked hemoglobin.
Claim 15
Independent Claim
82 words
A method of producing in commercially significant yields, a modified hemoglobin which is capable of functioning as a blood substitute and blood plasma expander characterized in that it comprises isolated unmodified hemoglobin or preparing a crude red cell lysate, deoxygenating the hemoglobin cross-linking the deoxygenated hemoglobin intramolecularly between Lys 99 Alpha 1 and Lys 99 Alpha 2 in the presence of an added polyanion selected from the group consisting of 2,3-diphosphoglycerate, inositol hexaphosphate, or inositol hexasulfate and purifying the alpha-alpha cross-linked hemoglobin.
Claim 16
Dependent Claim
13 words
A method according to claim 12, characterized in that the polyanion is 2,3-diphosphoglycerate.
Claim 17
Dependent Claim
14 words
A method according to claim 12, characterized in that the polyanion is inositol hexasulfate
Claim 18
Dependent Claim
14 words
A method according to claim 12, characterized in that the polyanion is inositol hexaphosphate.
Claim 19
Dependent Claim
25 words
A method according to claim 12, characterized in that the reaction is conducted at a temperature of from about 0°C to about 40 0 C.
Claim 20
Dependent Claim
23 words
A method according to claim 16, characterized in that the reaction is conducted at a temperature of from about 35°C to about 400C.
Claim 21
Dependent Claim
19 words
A method according to claim 16, characterized in that the reaction is conducted at a temperature of about 37°C.
Claim 22
Dependent Claim
30 words
A method according to claim 12, characterized in that the cross-linking between Lys 99 Alpha 1 and Lys 99 Alpha 2 is with a cross-linking agent which is amino group-specific
Claim 23
Dependent Claim
18 words
A method according to claim 19, characterized in that the cross-linking agent is a phenyl ester cross-linking agent.
Claim 24
Dependent Claim
15 words
A method according to claim 20, characterized in that the cross-linking agent is bis(3,5-dibromosalicyl) fumarate.
Claim 25
Dependent Claim
27 words
A method according to claim 19, characterized in that the amount of cross-linking agent employed is up to three fold molar excess of the amount of hemoglobin.
Claim 26
Dependent Claim
29 words
A method according to claim 22, characterized in that the amount of cross-linking agent is from about 1.3 times to about 2.0 times the molar amount of the hemoglobin.
Claim 27
Dependent Claim
34 words
A method according to claim 22, characterized in that the amount of the polyanion is from an amount equimolar to the amount of hemoglobin up to a twenty fold excess of the equimolar amount.
Claim 28
Dependent Claim
28 words
A method according to claim 24, characterized in that the amount of the polyanion is from about 5 to about 10 times the molar amount of the hemoglobin.
References Cited (Backward Citations)
No citation records indexed.
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