Reduced side-effect hemoglobin compositions

• US 6,670,323 B1
• Filed: 11/10/2000
• Issued: 12/30/2003
• Est. Priority Date: 11/12/1999
• Status: Expired due to Fees

• Current Assignee: Baxter Healthcare SA (Baxter International Inc.), Baxter International Inc.
• Original Assignee: Baxter Healthcare SA (Baxter International Inc.), Baxter International Inc.
• Inventors: Pereira, David, Doyle, Michael P., Burhop, Kenneth E., Hai, Ton-That, Hartman, James C., Lemon, Douglas D., Brucker, Eric A., Schick, Michael R., Moore, Edwin G., Apostol, Izydor Z., Trimble, Stephen P., Glascock, Christopher B., Lee, Geoffrey F., Foster, David L., Looker, Douglas L., Richards, Jane P.
• Primary Examiner(s): CARLSON, KAREN C
• Application Number:
  US09/709,914
• Time in Patent Office: 1,145 Days
• Field of Search: 514/6, 530/385
• US Class Current: 424/85.2
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/06 Antianaemics
• 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: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/805 Haemoglobins; Myoglobins

Abstract

The invention relates to novel hemoglobin compositions, particularly novel recombinant mutant hemoglobin compositions, which eliminate or substantially reduce 1) the creation of heart lesions, 2) gastrointestinal discomfort, 3) pressor effects, and 4) endotoxin hypersensitivity associated with the administration of extracellular hemoglobin compositions in various therapeutic applications. Applications described include treatments for anemia, head injury, hemorrhage or hypovolemia, ischemia, cachexia, sickle cell crisis and stroke; enhancing cancer treatments; stimulating hematopoiesis; improving repair of physically damaged tissues; alleviating cardiogenic shock; and shock resuscitation.

Claims

1. A hemoglobin comprising one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Tyr),

B10(Leu→

Trp),

B10(Leu→

TrP)+E11(Val→

Phe)+G8(Leu→

Ala),

B10(Leu→

Trp)+E7(His→

Gln),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val),

B10(Leu→

TrP)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val),

B10(Leu→

TrP)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val),

B10(Leu→

Phe)+E11(Val→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Trp),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile),

B10(Leu→

Phe)+E7(His→

Gln), and

B9(Ala→

Phe)

and/or said hemoglobin comprising one or more mutations selected from the group of beta globin mutations consisting of;

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp),

B9(Ala→

Leul+E11(Val→

Leu)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu),

B9(Ala→

Phe)+E11(Val→

Leu),

B9(Ala→

TrP)+E11(Val→

Leu),

B9(Ala→

Leu)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Phe).

B9(Ala→

Phe)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Trp),

B9(Ala→

Phe),

B9(Ala→

Trp),

B9(Ala→

Leu),

B9(Ala→

Met),

B10(Leu→

Phe),

B10(Leu→

Phe)+B14(Leu→

Phe),

B10(Leu→

Phe)+E4(Val→

Trp)

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp),

E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Met),

E11(Val→

Met)+G8(Leu→

Phe),

E11(Val→

Trp)+G8(Leu→

Trp),

EF6(Lys→

Asp),

G8(Leu→

Trp), and

G8(Leu→

Phe).

2. The hemoglobin of claim 1 wherein the hemoglobin is reduced gastrointestinal effect hemoglobin.

3. The hemoglobin of claim 1 wherein the hemoglobin is low gastrointestinal effect hemoglobin.

4. The hemoglobin of claim 1 wherein the hemoglobin is low pressor effect hemoglobin.

5. The hemoglobin of claim 1 wherein the hemoglobin exhibits decreased aggregate formation when stored in solution in deoxygenated form for prolonged periods.

6. The hemoglobin of claim 5, wherein the hemoglobin further comprises a mutation in a beta globin at F3Thr.

7. The hemoglobin of claim 6 wherein the hemoglobin further comprises a mutation chosen from the group consisting of F3 Thr→

Asp, Glu, Arg, Lys, His, Tyr, Gln, and Trp.

8. The hemoglobin of claim 7 wherein the hemoglobin further comprises a mutation chosen from the group consisting of F3 Thr→

Asp, Gln, Glu, Arg, Lys, and His.

9. The hemoglobin of claim 8 wherein the mutation comprises F3 Thr→

Gln.

10. The hemoglobin of claim 1 wherein the hemoglobin is reduced lesion hemoglobin.

12. The hemoglobin of claim 10 or 11 wherein the hemoglobin is a reduced pressor effect hemoglobin.

13. The hemoglobin of claim 11, 10 or 12 wherein the hemoglobin is reduced gastrointestinal effect hemoglobin.

14. The hemoglobin of claim 11, 10 or 12 wherein the hemoglobin is low gastrointestinal effect hemoglobin.

15. The hemoglobin of claim 11, 10 or 12 wherein the hemoglobin exhibits decreased aggregate formation when stored in solution in deoxygenated form for prolonged periods.

16. The hemoglobin of claim 15 wherein the hemoglobin further comprises a mutation in a beta globin at F3Thr.

17. The hemoglobin of claim 16 wherein the hemoglobin further comprises a mutation chosen from the group consisting of F3 Thr→

Asp, Glu, Arg, Lys, His, Tyr, Gln, and Trp.

18. The hemoglobin of claim 17 wherein the hemoglobin further comprises a mutation chosen from the group consisting of F3 Thr→

Asp, Gln, Glu, Arg, Lys, and His.

19. The hemoglobin of claim 18 wherein the mutation comprises F3 Thr→

Gln.

20. The hemoglobin of claim 10 or 11 wherein the hemoglobin is a low pressor effect hemoglobin.

21. The hemoglobin of claim 10 or 11, wherein said hemoglobin is a reduced endotoxin effect hemoglobin.

22. The hemoglobin of claim 12 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 130 to 500 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

23. The hemoglobin of claim 12 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 190 to 350 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises three or more hemoglobin-like proteins.

24. The hemoglobin of claim 12 wherein the hemoglobin includes about 40 to about 60% derivatized monomeric hemoglobin with an average molecular weight between 65 and less than about 130 kilodaltons, and about 40 to about 60% oligomeric hemoglobin with an average molecular weight between 130 and 650 kilodaltons, wherein the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

25. The hemoglobin of claim 1 wherein the hemoglobin reduces the rate of NO scavenging.

26. The hemoglobin of claim 25 wherein the hemoglobin has nitric oxide kinetics of less than 10 μ

M<sup>−

1</sup>s<sup>−

1</sup>.

27. The hemoglobin of claim 1 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than 50% in the range of 130 to 500 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

28. The hemoglobin of claim 1 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than 50% in the range of 190 to 350 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises three or more hemoglobin-like proteins.

29. The hemoglobin of claim 1 wherein the hemoglobin has an average molecular weight between 65 and 650 kilodaltons.

30. The hemoglobin of claim 1 wherein the hemoglobin includes 40 to 60% derivatized monomeric hemoglobin with an average molecular weight between 65 and 130 kilodaltons, and 40 to 60% oligomeric hemoglobin with an average molecular weight between 130 and 650 kilodaltons, wherein the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

31. The hemoglobin of claim 1, wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

[B10(Leu-Trp)+E7(His-Gln),

B10(Leu-Phe),

B10(Leu-Trp), and

B10(Leu-Tyr)]

B10(Leu→

Tyr),

B10(Leu→

Trp), and

B10(Leu→

Trp)+E7(His→

Gln)

and/or wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of;

E11(Val→

Met)+G8(Leu→

Phe),

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp), and

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp).

32. The hemoglobin of claim 1 wherein the hemoglobin comprises the alpha globin mutations B10(Leu→

Trp)+E7(His→

Gln).

33. The hemoglobin of claim 1 wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Tyr);

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe); and

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile);

or B9(Ala→

Phe).

34. The hemoglobin of claim 1 wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of:

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu);

B9(Ala→

Phe)+E11(Val→

Leu);

B9(Ala→

Trp)+E11(Val→

Leu);

B9(Ala→

Phe);

B9(Ala→

Trp);

B9(Ala→

Leu);

B9(Ala→

Met);

E11(Val→

Met);

B10(Leu→

Phe)+B14(Leu→

Phe);

B9(Ala→

Leu)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Phe);

B9(Ala→

Phe)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Trp);

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp); and

E11(Val→

Trp)+G8(Leu→

Trp).

52. The hemoglobin of claim 1 wherein the hemoglobin is polymerized.

11. A hemoglobin, said hemoglobin being polymerized and being lesion free, said hemoglobin comprising one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Phe),

B10(Leu→

Tyr),

B10(Leu→

Trp),

B10(Leu→

Trp)+E11(Val→

Phe),

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala),

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ile),

B10(Leu→

Trp)+E11(Val→

Leu)+G8(Leu→

Phe),

B10(Leu→

Trp)+E11(Val→

Trp),

B10(Leu→

Trp)+E11(Val→

Trp)+G8(Leu→

Trp),

B10(Leu→

Trp)+E7(His→

Gln),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Trp),

B10(Leu→

Phe)+E11(Val→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Trp),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile),

B10(Leu→

Phe)+E7(His→

Gln),

E11(Val→

Leu),

E11(Val→

Phe),

E11(Val→

Trp), and

B9(Ala→

Phe)

and/or said hemoglobin comprising one or more mutations selected from the group of beta globin mutations consisting of;

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu),

B9(Ala→

Phe)+E11(Val→

Leu),

B9(Ala→

Trp)+E11(Val→

Leu),

B9(Ala→

Leu)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Phe),

B9(Ala→

Phe)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Trp),

B9(Ala→

Phe),

B9(Ala→

Trp),

B9(Ala→

Leu),

B9(Ala→

Met),

B10(Leu→

Phe),

B10(Leu→

Phe)+B14(Leu→

Phe),

B10(Leu→

Phe)+E4(Val→

Leu),

B10(Leu→

Phe)+E4(Val→

Trp),

B10(Leu→

Trp)+E4(Val→

Leu),

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp), and

B13(Leu→

Met)+E11(Val→

Trp),

E7(His→

Gln)+E11(Val→

Trp),

E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Met),

E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Met)+G8(Leu→

Phe),

E11(Val→

Trp)

E11(Val→

Trp)+G8(Leu→

Trp),

E11(Val→

Phe),

E11(Val→

Phe)+G8(Leu→

Phe),

E11(Val→

Leu),

E11(Val→

Leu)+G8(Leu→

Phe),

E11(Val→

Leu)+G8(Leu→

Trp),

EF6(Lys→

Asp),

G8(Leu→

Trp),

G8(Leu→

Trp)+G12(Leu→

Trp),

G8(Leu→

Phe), and

G8(Leu→

Phe)+G12(Leu→

Trp).

35. The hemoglobin of claim 11 wherein the hemoglobin reduces the rate of NO scavenging.

36. The hemoglobin of claim 35 wherein the hemoglobin has nitric oxide kinetics of less than 10 μ

M<sup>−

1</sup>s<sup>−

1</sup>.

37. The hemoglobin of claim 11 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than 50% in the range of 130 to 500 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

38. The hemoglobin of claim 11 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than 50% in the range of 190 to 350 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises three or more hemoglobin-like proteins.

39. The hemoglobin of claim 11 wherein the hemoglobin has an average molecular weight between 65 and 650 kilodaltons.

40. The hemoglobin of claim 11 wherein the hemoglobin includes 40 to 60% derivatized monomeric hemoglobin with an average molecular weight between 65 and 130 kilodaltons, and 40 to 60% oligomeric hemoglobin with an average molecular weight between 130 and 650 kilodaltons, wherein the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

41. The hemoglobin of claim 11 wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu-Trp)+E7(His-Gln),

B10(Leu-Phe),

B10(Leu-Trp), and

B10(Leu-Tyr)

and/or wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of;

E11(Val→

Trp),

E11(Val→

Met)+G8(Leu→

Phe),

E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Leu)+G8(Leu→

Trp),

G8(Leu→

Phe)+G12(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp), and

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp).

42. The hemoglobin of claim 11 wherein the hemoglobin comprises the alpha globin mutations B10(Leu→

Trp)+E7(His→

Gln).

43. The hemoglobin of claim 11 wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Tyr);

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe); and

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile);

or B9(Ala→

Phe).

44. The hemoglobin of claim 11 wherein the hemoglobin comprises the beta globin mutation E11(Val→

Trp).

45. The hemoglobin of claim 11 wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of:

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu);

B9(Ala→

Phe)+E11(Val Leu);

B9(Ala→

Trp)+E11(Val→

Leu);

B9(Ala→

Phe);

B9(Ala→

Trp);

B9(Ala→

Leu);

B9(Ala→

Met);

E11(Val→

Met);

B10(Leu→

Phe)+B14(Leu→

Phe);

B9(Ala→

Leu)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Phe);

B9(Ala→

Phe)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Trp);

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp); and

E11(Val→

Trp)+G8(Leu→

Trp).

53. The hemoglobin of claim 11 wherein the hemoglobin is polymerized.

46. A hemoglobin comprising a combination of mutations in the alpha and beta subunits selected from the group mutations consisting of:

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ala);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ser);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Met);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

FG1(Asp→

Asn);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

HC3(His→

Gln);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

NA2(His→

Arg);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G4(Asn→

Tyr);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G10(Asn→

Lys);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp); and

α

₁B10(Leu<

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln)+β

E11(Val→

Met)+β

G8(Leu→

Trp).

47. A hemoglobin comprising one or more mutations selected from the group of delta globin mutations consisting of:

E11(Val→

Ttp),

E11(Val→

Trp)+EF6(Lys→

Asp),

A6(Thr→

Cys)+E11(Val→

Trp),

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp), and

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp)+F9(Lys→

Ala).

48. A hemoglobin comprising one or more mutations selected from the group of dialpha mutations consisting of:

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln),

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp),

α

₁B10(Leu→

Phe)+(Gly, Gly)+α

₂B10(Leu→

Trp), and

α

₁B10(Leu→

Phe)+(Gly, Gly, Gly)+α

₂B10(Leu→

Trp).

49. A hemoglobin being polymerized and being lesion free, said hemoglobin comprising a combination of mutations in the alpha and beta subunits selected from the group mutations consisting of:

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ala);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ser);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Met);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

FG1(Asp→

Asn);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

HC3(His→

Gln);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

NA2(His→

Arg);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G4(Asn→

Tyr);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G10(Asn→

Lys);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp); and

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln)+β

E11(Val→

Met)+β

G8(Leu→

Trp).

50. A hemoglobin being polymerized and being lesion free, said hemoglobin comprising one or more mutations selected from the group of delta globin mutations consisting of:

E11(Val→

Trp),

E11(Val→

Trp)+EF6(Lys→

Asp),

A6(Thr→

Cys)+E11(Val→

Trp),

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp), and

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp)+F9(Cys→

Ala).

51. A hemoglobin being polymerized and being lesion free, said hemoglobin comprising one or more mutations selected from the group of dialpha mutations consisting of:

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln),

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp),

α

₁B10(Leu→

Phe)+(Gly, Gly)+α

₂B10(Leu→

Trp), and

α

₁B10(Leu→

Phe)+(Gly, Gly, Gly)+α

₂B10(Leu→

Trp).

54. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of hemoglobin per deciliter of pharmaceutically acceptable carrier, said hemoglobin comprising one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Tyr),

B10(Leu→

Trp),

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala),

B10(Leu→

Trp)+E7(His→

Gln),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val),

B10(Leu→

Phe)+E11(Val→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Trp),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile),

B10(Leu→

Phe)+E7(His→

Gln), and

B9(Ala→

Phe)

and/or said hemoglobin comprising one or more mutations selected from the group of beta globin mutations consisting of;

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu),

B9(Ala→

Phe)+E11(Val→

Leu),

B9(Ala→

Trp)+E11(Val→

Leu),

B9(Ala→

Leu)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Phe),

B9(Ala→

Phe)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Trp),

B9(Ala→

Phe),

B9(Ala→

Trp),

B9(Ala→

Leu),

B9(Ala→

Met),

B10(Leu→

Phe),

B10(Leu→

Phe)+B14(Leu→

Phe),

B10(Leu→

Phe)+E4(Val→

Trp),

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp),

E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Met),

E11(Val→

Met)+G8(Leu→

Phe),

E11(Val→

Trp)+G8(Leu→

Trp),

EF6(Lys→

Asp),

G8(Leu→

Trp), and

G8(Leu→

Phe).

55. The pharmaceutical composition of claim 54 wherein the hemoglobin is reduced gastrointestinal effect hemoglobin.

56. The pharmaceutical composition of claim 55 wherein the hemoglobin is reduced lesion hemoglobin.

57. The pharmaceutical composition of claim 54 wherein the hemoglobin is low gastrointestinal effect hemoglobin.

58. The pharmaceutical composition of claim 54 wherein the hemoglobin is low pressor effect hemoglobin.

59. The pharmaceutical composition of claim 58, wherein the hemoglobin is polymerized.

60. The pharmaceutical composition of claim 54 wherein the hemoglobin exhibits decreased aggregate formation when stored in solution in deoxygenated form for prolonged periods.

61. The pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of the hemoglobin of claim 60, wherein said hemoglobin further comprises a mutation in a beta globin at F3Thr.

62. The pharmaceutical composition of claim 61, wherein the amino acid at F3Thr is selected from the group consisting of Asp, Glu, Arg, Lys, His, Tyr, Gln, and Trp.

63. The pharmaceutical composition of claim 62, wherein the amino acid at F3Thr is selected the group consisting of Asp, Gln, Glu, Arg, Lys, and His.

64. The pharmaceutical composition of claim 63, wherein the amino acid at F3Thr is Gln.

65. The pharmaceutical composition of claim 54 wherein the hemoglobin is reduced lesion hemoglobin.

68. The pharmaceutical composition of claim 65 or 66 wherein the hemoglobin is low gastrointestinal effect hemoglobin.

69. The pharmaceutical composition of claim 65 or 66 wherein the hemoglobin is reduced pressor effect hemoglobin.

70. The pharmaceutical composition of claim 69 wherein the hemoglobin is reduced gastrointestinal effect hemoglobin.

71. The pharmaceutical composition of claim 69 wherein the hemoglobin is low gastrointestinal effect hemoglobin.

72. The pharmaceutical composition of claim 69 wherein the hemoglobin exhibits decreased aggregate formation when stored in solution in deoxygenated form for prolonged periods.

73. The pharmaceutical composition of claim 72, wherein said hemoglobin further comprises a mutation in a beta globin at F3Thr.

74. The pharmaceutical composition of claim 73, wherein the amino acid at F3Thr is selected from the group consisting of Asp, Glu, Arg, Lys, His, Tyr, Gln, and Trp.

75. The pharmaceutical composition of claim 74, wherein the amino acid at F3Thr is selected from the group consisting of Asp, Gln, Glu, Arg, Lys, and His.

76. The pharmaceutical composition of claim 75, wherein the amino acid at F3Thr is Gln.

77. The pharmaceutical composition of claim 69 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 130 to 500 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

78. The pharmaceutical composition of claim 69 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 190 to 350 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises three or more hemoglobin-like proteins.

79. The pharmaceutical composition of claim 69 wherein the hemoglobin includes about 40 to about 60% derivatized monomeric hemoglobin with an average molecular weight between 65 and less than about 130 kilodaltons, and about 40 to about 60% oligomeric hemoglobin with an average molecular weight between 130 and 650 kilodaltons, wherein the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

80. The pharmaceutical composition of claim 65 or 66 wherein the hemoglobin exhibits decreased aggregate formation when stored in solution in deoxygenated form for prolonged periods.

81. The pharmaceutical composition of claim 65 or 66 wherein the hemoglobin is low pressor effect hemoglobin.

82. The pharmaceutical composition of claim 65 or 66 wherein the hemoglobin is reduced endotoxin hemoglobin.

83. The pharmaceutical composition of claim 54 wherein the hemoglobin reduces the rate of NO scavenging.

84. The pharmaceutical composition of claim 83 wherein the hemoglobin has nitric oxide kinetics of less than about 10 μ

M<sup>−

1</sup>s<sup>−

1</sup>.

85. The pharmaceutical composition of claim 54 wherein the composition:

a) is substantially free of red blood cell stroma, b) is substantially free of pyrogens, and c) comprises less than about 10% methemoglobin.

86. The pharmaceutical composition of claim 54 wherein the composition is further characterized in that the composition has a P₅₀ measured under human physiologic conditions which is greater than that of cell free human hemoglobin A_o.

87. The pharmaceutical composition of claim 54 wherein the composition is further characterized in that the composition has a P₅₀ measured under human physiologic conditions which is between 20 and 50.

88. The pharmaceutical composition of claim 54 wherein the composition is further characterized in that the composition has a P₅₀ measured under human physiologic conditions which is between 25 and 45.

89. The pharmaceutical composition of claim 54 wherein the hemoglobin has an average molecular weight between 65 and 650 kilodaltons.

90. The pharmaceutical composition of claim 54 wherein the hemoglobin includes 40 to 60% derivatized monomeric hemoglobin with an average molecular weight between 65 and 130 kilodaltons, and 40 to 60% oligomeric hemoglobin with an average molecular weight between 130 and 650 kilodaltons, wherein the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

91. The pharmaceutical composition of claim 54 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 130 to 500 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

92. The pharmaceutical composition of claim 54 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 190 to 350 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises three or more hemoglobin-like proteins.

93. The pharmaceutical composition of claim 54, wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

[B10(Leu-Trp)+E7(His-Gln),

B10(Leu-Phe),

B10(Leu-Trp), and

B10(Leu-Tyr)]

B10(Leu→

Tyr),

B10(Leu→

Trp), and

B10(Leu→

Trp)+E7(His→

Gln)

and/or wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of;

E11(Val→

Met)+G8(Leu→

Phe),

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp), and

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp).

94. The pharmaceutical composition of claim 54 wherein the hemoglobin comprises the alpha globin mutations B10(Leu→

Trp)+E7(His→

Gln).

95. The pharmaceutical composition of claim 54 wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Tyr);

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe); and

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile);

or B9(Ala→

Phe).

96. The pharmaceutical composition of claim 54 wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of:

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu);

B9(Ala→

Phe)+E11(Val→

Leu);

B9(Ala→

Trp)+E11(Val→

Leu);

B9(Ala→

Phe);

B9(Ala→

Trp);

B9(Ala→

Leu);

B9(Ala→

Met);

E11(Val→

Met);

B10(Leu→

Phe)+B14(Leu→

Phe);

B9(Ala→

Leu)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Phe);

B9(Ala→

Phe)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Trp);

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp); and

E11(Val→

Trp)+G8(Leu→

Trp).

66. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of hemoglobin per deciliter of pharmaceutically acceptable carrier, said hemoglobin being polymerized and being lesion free, said hemoglobin comprising one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Phe),

B10(Leu→

Tyr),

B10(Leu→

Trp),

B10(Leu→

Trp)+E11(Val→

Phe),

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala),

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ile),

B10(Leu→

Trp)+E11(Val→

Leu)+G8(Leu→

Phe),

B10(Leu→

Trp)+E11(Val→

Trp),

B10(Leu→

Trp)+E11(Val→

Trp)+G8(Leu→

Trp),

B10(Leu→

Trp)+E7(His→

Gln),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val),

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Trp),

B10(Leu→

Phe)+E11(Val→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Trp),

B10(Leu→

Phe)+E7(His→

Gln)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met),

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile),

B10(Leu→

Phe)+E7(His→

Gln),

E11(Val→

Leu),

E11(Val→

Phe),

E11(Val→

Trp), and

B9(Ala→

Phe)

and/or said hemoglobin comprising one or more mutations selected from the group of beta globin mutations consisting of;

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Leu),

B9(Ala→

Phe)+E11(Val→

Leu),

B9(Ala→

Trp)+E11(Val→

Leu),

B9(Ala→

Leu)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Phe),

B9(Ala→

Phe)+G8(Leu→

Trp),

B9(Ala→

Trp)+G8(Leu→

Trp),

B9(Ala→

Phe),

B9(Ala→

Trp),

B9(Ala→

Leu),

B9(Ala→

Met),

B10(Leu→

Phe),

B10(Leu→

Phe)+B14(Leu→

Phe),

B10(Leu→

Phe)+E4(Val→

Leu),

B10(Leu→

Phe)+E4(Val→

Trp),

B10(Leu→

Trp)+E4(Val→

Leu),

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp), and

B13(Leu→

Met)+E11(Val→

Trp),

E7(His→

Gln)+E11(Val→

Trp),

E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Met),

E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Met)+G8(Leu→

Phe),

E11(Val→

Trp)

E11(Val→

Trp)+G8(Leu→

Trp),

E11(Val→

Phe),

E11(Val→

Phe)+G8(Leu→

Phe),

E11(Val→

Leu),

E11(Val→

Leu)+G8(Leu→

Phe),

E11(Val→

Leu)+G8(Leu→

Trp),

EF6(Lys→

Asp),

G8(Leu→

Trp),

G8(Leu→

Trp)+G12(Leu→

Trp),

G8(Leu→

Phe), and

G8(Leu→

Phe)+G12(Leu→

Trp).

67. The pharmaceutical composition of claim 66 wherein the hemoglobin is reduced gastrointestinal effect hemoglobin.

97. The pharmaceutical composition of claim 66 wherein the hemoglobin reduces the rate of NO scavenging.

98. The pharmaceutical composition of claim 97 wherein the hemoglobin has nitric oxide kinetics of less than about 10 μ

M<sup>−

1</sup>s<sup>−

1</sup>.

99. The pharmaceutical composition of claim 66 wherein the composition:

a) is substantially free of red blood cell stroma, b) is substantially free of pyrogens, and c) comprises less than about 10% methemoglobin.

100. The pharmaceutical composition of claim 66 wherein the composition is further characterized in that the composition has a P₅₀ measured under human physiologic conditions which is greater than that of cell free human hemoglobin A_o.

101. The pharmaceutical composition of claim 66 wherein the composition is further characterized in that the composition has a P₅₀ measured under human physiologic conditions which is between 20 and 50.

102. The pharmaceutical composition of claim 66 wherein the composition is further characterized in that the composition has a P₅₀ measured under human physiologic conditions which is between 25 and 45.

103. The pharmaceutical composition of claim 66 wherein the hemoglobin has an average molecular weight between 65 and 650 kilodaltons.

104. The pharmaceutical composition of claim 66 wherein the hemoglobin includes 40 to 60% derivatized monomeric hemoglobin with an average molecular weight between 65 and 130 kilodaltons, and 40 to 60% oligomeric hemoglobin with an average molecular weight between 130 and 650 kilodaltons, wherein the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

105. The pharmaceutical composition of claim 66 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 130 to 500 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises two or more hemoglobin-like proteins.

106. The pharmaceutical composition of claim 66 wherein the hemoglobin is an oligomeric hemoglobin with a molecular weight distribution of greater than about 50% in the range of 190 to 350 kilodaltons, and wherein at least 80% of the oligomeric hemoglobin comprises three or more hemoglobin-like proteins.

107. The pharmaceutical composition of claim 66, wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu-Trp)+E7(His-Gln),

B10(Leu-Phe),

B10(Leu-Trp), and

B10(Leu-Tyr)

and/or wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of;

E11(Val→

Trp),

E11(Val→

Met)+G8(Leu→

Phe),

E11(Val→

Met)+G8(Leu→

Trp),

E11(Val→

Leu)+G8(Leu→

Trp),

G8(Leu→

Phe)+G12(Leu→

Trp),

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp), and

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp).

108. The pharmaceutical composition of claim 66 wherein the hemoglobin comprises the alpha globin mutations B10(Leu→

Trp)+E7(His→

Gln).

109. The pharmaceutical composition of claim 66 wherein the hemoglobin comprises one or more mutations selected from the group of alpha globin mutations consisting of:

B10(Leu→

Tyr);

B10(Leu→

Trp)+E11(Val→

Phe)+G8(Leu→

Ala);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Phe)+G8(Leu→

Val);

B10(Leu→

Trp)+E7(His→

Gln)+E11(Val→

Leu)+G8(Leu→

Val);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Phe); and

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Met);

B10(Leu→

Phe)+E7(His→

Gln)+E11(Val→

Met)+G8(Leu→

Ile);

or B9(Ala→

Phe).

110. The pharmaceutical composition of claim 66 wherein the hemoglobin comprises the beta globin mutation E11(Val→

Trp).

111. The pharmaceutical composition of claim 66 wherein the hemoglobin comprises one or more mutations selected from the group of beta globin mutations consisting of:

B9(Ala→

Leu)+E11(Val→

Met)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu)+G8(Leu→

Trp);

B9(Ala→

Leu)+E11(Val→

Leu);

B9(Ala→

Phe)+E11(Val→

Leu);

B9(Ala→

Trp)+E11(Val→

Leu);

B9(Ala→

Phe);

B9(Ala→

Trp);

B9(Ala→

Leu);

B9(Ala→

Met);

E11(Val→

Met);

B10(Leu→

Phe)+B14(Leu→

Phe);

B9(Ala→

Leu)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Phe);

B9(Ala→

Phe)+G8(Leu→

Trp);

B9(Ala→

Trp)+G8(Leu→

Trp);

B10(Leu→

Ala)+E7(His→

Phe)+G8(Leu→

Trp); and

E11(Val→

Trp)+G8(Leu→

Trp).

112. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of reduced lesion hemoglobin per deciliter of pharmaceutically acceptable carrier, wherein the hemoglobin comprises a combination of mutations in the alpha and beta subunits selected from the group mutations consisting of:

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ala);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ser);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Met);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

FG1(Asp→

Asn);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

HC3(His→

Gln);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

NA2(His→

Arg);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G4(Asn→

Tyr);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G10(Asn→

Lys);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp); and

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln)+β

E11(Val→

Met)+β

G8(Leu→

Trp).

119. A method for increasing the delivery of oxygen to living cells within a mammal comprising administering to the mammal an oxygen delivery assisting amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 61.

120. The method according to claim 119 wherein the oxygen delivery assisting amount of the pharmaceutical composition is between 1 mg and 15,000 mg per kilogram body weight.

121. The method according to claim 119 wherein the oxygen delivery assisting amount of the pharmaceutical composition is between 100 mg and 7,500 mg per kilogram body weight.

122. The method according to claim 119 wherein the oxygen delivery assisting amount of the pharmaceutical composition is between 500 mg and 5,000 mg per kilogram body weight.

123. The method according to claim 119 wherein the oxygen delivery assisting amount of the pharmaceutical composition is between 700 mg and 3,000 mg per kilogram body weight.

124. A method for priming a surgical device comprising filling a fluid void volume of the surgical device with the pharmaceutical composition of any one of claims 54, 66, 112, 115, 113, and 114.

125. The method of claim 124 wherein the surgical device is selected from a group consisting of an angioplasty catheter, a blood oxygenator, and a heart-lung machine.

126. A method of effecting hemodilution of a patient'"'"'s blood comprising the administration of a hemodilution effecting amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

127. The method of claim 126 wherein the patient is hemodiluted to a hypervolemic state.

128. The method according to claim 126 wherein the hemodilution is carried out perioperatively.

129. The method according to claim 126 wherein the hemodilution effecting amount of the pharmaceutical composition is between 1 ml and 100 ml per kilogram of body weight.

130. A method for treating hypovolemia in a mammal comprising administering to the mammal a hypovolemia alleviating amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

131. The method of claim 130 wherein the hypovolemia is the result of hemorrhage.

132. The method of claim 130 wherein the hypovolemia is accompanied by hypovolemic shock.

133. The method of claim 130 wherein the pharmaceutical composition is administered within the three hours of hypovolemic onset.

134. The method of claim 130 wherein the pharmaceutical composition is administered within the first hour of hypovolemic onset.

135. The method of claim 130 wherein the hypovolemia alleviating amount of pharmaceutical composition is administered as a single intravenous injection.

136. The method of claim 130 wherein the hypovolemia alleviating amount of pharmaceutical composition is administered as an intravenous infusion.

137. A method for treating cardiogenic shock in a mammal comprising administering to the mammal a shock alleviating amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

138. The method of claim 137 wherein the shock alleviating amount is from about 1 ml to about 6000 ml per kilogram of body weight.

139. The method of claim 138 wherein the pharmaceutical composition is administered within the first hour of cardiogenic shock.

140. A method for resuscitating a mammal in a shock state comprising administering to the mammal a resuscitating amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

141. The method of claim 140 wherein the shock state is selected from the group consisting of hypovolemic shock, hemorrhagic shock, and cardiogenic shock.

142. A method for increasing tissue perfusion in a mammal comprising administering to the mammal a tissue perfusion increasing amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113 and 114.

143. The method of claim 142 wherein the mammal is suffering from a stroke.

144. The method of claim 142 wherein the mammal has tissue in an ischemic state.

145. The method of claim 142 wherein the mammal has tissue which has suffered physical damage.

146. The method of claim 142 wherein the mammal has a burn or wound.

147. A method for stimulating hematopoiesis in a mammal comprising administering to the mammal a hematopoiesis stimulating amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

148. The method of claim 147 further comprising the co-administration of one or more other hematopoietic factors.

149. The method of claim 148 wherein the hematopoietic factors are chosen from the group consisting of GM-CSF, M-CSF, G-CSF, SCF, EPO, AND IL1 through IL13.

150. A method for treating a cytopenia in a mammal comprising administering to the mammal a cytopenia alleviating amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

151. The method of claim 150 wherein the cytopenia is an anemia.

152. The method of claim 150 wherein the cytopenia is an AIDS or AZT therapy-related anemia.

153. A method for treating cachexia in a mammal comprising administering to the mammal a cachexia alleviating amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

154. A method for treating a head injury in a mammal comprising administering to the mammal an intracranial pressure relieving amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

155. A method for enhancing a cancer therapy in a mammal comprising administering to the mammal, prior to or about contemporaneously with the therapy, a therapy enhancing amount of the pharmaceutical composition of of any one of claims 54, 66, 112, 115, 113, and 114.

113. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of reduced lesion hemoglobin per deciliter of pharmaceutically acceptable carrier, wherein the hemoglobin comprises one or more mutations selected from the group of delta globin mutations consisting of:

E11(Val→

Trp),

E11(Val→

Trp)+EF6(Lys→

Asp),

A6(Thr→

Cys)+E11(Val→

Trp),

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp), and

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp)+F9(Lys→

Ala).

118. The pharmaceutical composition of any one of claims 54, 66, 113, and 61, wherein the hemoglobin is polymerized.

114. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of reduced lesion hemoglobin per deciliter of pharmaceutically acceptable carrier, wherein the hemoglobin comprises one or more mutations selected from the group of dialpha mutations consisting of:

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln),

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp),

α

₁B10(Leu→

Phe)+(Gly, Gly)+α

₂B10(Leu→

Trp), and

α

₁B10(Leu→

Phe)+(Gly, Gly, Gly)+α

₂B10(Leu→

Trp).

115. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of hemoglobin per deciliter of pharmaceutically acceptable carrier, said hemoglobin being polymerized and being lesion free, wherein the hemoglobin comprises a combination of mutations in the alpha and beta subunits selected from the group mutations consisting of:

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ala);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Ser);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

F9(Cys→

Met);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

FG1(Asp→

Asn);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

HC3(His→

Gln);

α

B10(Leu→

Trp)+α

E7(His→

Gln)+β

E11(Val→

Trp)+β

NA2(His→

Arg);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G4(Asn→

Tyr);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

G10(Asn→

Lys);

α

B10(Leu→

Phe)+β

E11(Val→

Trp)+β

EF6(Lys→

Asp); and

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln)+β

E11(Val→

Met)+β

G8(Leu→

Trp).

116. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of hemoglobin per deciliter of pharmaceutically acceptable carrier, said hemoglobin being polymerized and being lesion free, wherein the hemoglobin comprises one or more mutations selected from the group of delta globin mutations consisting of:

E11(Val→

Trp),

E11(Val→

Trp)+EF6(Lys→

Asp),

A6(Thr→

Cys)+E11(Val→

Trp),

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp), and

A6(Thr→

Cys)+E11(Val→

Trp)+EF6(Lys→

Asp)+F9(Cys→

Ala).

117. A pharmaceutical composition for transporting and releasing oxygen comprising a suspension from 5 to 20 grams of hemoglobin per deciliter of pharmaceutically acceptable carrier, said hemoglobin being polymerized and being lesion free, wherein the hemoglobin comprises one or more mutations selected from the group of dialpha mutations consisting of:

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp)+α

₂E7(His→

Gln),

α

₁B10(Leu→

Phe)+α

₂B10(Leu→

Trp),

α

₁B10(Leu→

Phe)+(Gly, Gly)+α

₂B10(Leu→

Trp), and

α

₁B10(Leu→

Phe)+(Gly, Gly, Gly)+α

₂B10(Leu→

Trp).