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Modified human hemoglobin, blood substitutes containing the same, and vectors for expressing the modified hemoglobin
| Details |
Inventors: Fronticelli, Clara; Bucci, Enrico; Brinigar, William;
Assignee: Research Corporation Technologies, Inc. (Tucson, AZ)
Primary Examiner: Nucker; Christine M.
Assistant Examiner: Sidberry; H. F.
Attorney, Agent or Firm: Birch, Stewart, Kolasch & Birch
The present invention relates to modified .beta.-chains of human hemoglobin having an oxygen binding affinity equal to or less than natural human hemoglobin. The modified the .beta.-chain of human hemoglobin of the invention preferably has at least one chloride binding site not present in .beta.-chain of natural human hemoglobin. |
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DETAILED DESCRIPTION The present invention relates to modified .
beta.
-chains of human hemoglobin having an oxygen binding affinity equal to or less than natural human hemoglobin.
Preferably, the oxygen binding affinity P.
sub.
50 of the modified .
beta.
-chains is in the range of about 3 to 70 mmHG as measured at temperatures in the range of about 10.
degree.
to 40.
degree.
C.
, at pH 7.
4, in the presence of 0.
1 to 0.
2 M chloride.
The modified the .
beta.
-chain of human hemoglobin of the invention preferably has at least one chloride binding site not present in the .
beta.
-chain of natural human hemoglobin.
Also, the modified .
beta.
-chain of human hemoglobin of the invention has additional negative charges at the N-terminal residues of the .
beta.
-chains.
Moreover, the modified .
beta.
-chain of human hemoglobin of the invention may have hydrophobic residues, (e.
g.
, Ala, Gly, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr or Val) at the N-terminal residues of the .
beta.
-chains.
Representative examples of the modified .
beta.
-chain of human hemoglobin include the following site specific mutants: 1) NA1 (1) Val.
fwdarw.
Met; delete NA2 (2) His.
2) HCl (144) Lys.
fwdarw.
Arg.
3) NA1 (1) Val.
fwdarw.
Met; NA2 (2) His deleted; HCl (144) Lys.
fwdarw.
Arg.
4) NA1(1) Val.
fwdarw.
Met; NA2 (2) His deleted; E20 (76)Ala.
fwdarw.
Lys.
5) NA1(1) Val.
fwdarw.
Met; NA2 (2) His deleted; E20 (76) Ala.
fwdarw.
Lys; HCl (144) Lys.
fwdarw.
Arg.
6) NA3 (3) Leu.
fwdarw.
Glu or Asp.
7) Al (4) Thr.
fwdarw.
Glu or Asp.
8) A2 (5) Pro.
fwdarw.
Asp or Glu.
9) A2 (5) Pro.
fwdarw.
Gly, Ala or an amino acid having greater backbone flexibility.
10) E13 (69) Gly.
fwdarw.
Glu or Asp.
11) E14 (70) Ala.
fwdarw.
Thr or Ser.
12) E19 (75) Leu.
fwdarw.
Cyst or Met.
13) E20 (76) Ala.
fwdarw.
Arg, His, or Lys.
14) NA1 (1) Val.
fwdarw.
Met or Leu.
15) NA2 (2) His.
fwdarw.
Leu, Tyr or Asp.
16) NA1 (1) Val, NA2 (2) Hist.
fwdarw.
Met-Leu, Met-Asp, Met-Tyr, Leu-Leu, Leu-Asp or Leu-Tyr.
17) NA1 (1) Val.
fwdarw.
(Gly).
sub.
n - Cys, where n is 1 to 20.
18) E11 (67) Val.
fwdarw.
Thr.
The number in the parenthesis in the above list of mutants represents the sequence number of the amino acids in the
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Drugs 
