Claims
- 1. An oxidation-resistant IL-2 mutein having therapeutically useful biological activity, wherein each methionine residue of a reference protein susceptible to chloramine T or peroxide oxidation is replaced by a conservative amino acid, and wherein additional, non-susceptible methionine residues are not so substituted.
- 2. The oxidation-resistant mutein of claim 1 wherein the conservative amino acid is selected from the group consisting of glycine, alanine, serine, threonine, valine, isoleucine, leucine, asparagine, glutamine, glutamate, tyrosine, and phenylalanine.
- 3. The oxidation-resistant mutein of claim 2 wherein the reference protein is human.
- 4. The oxidation-resistant mutein of claim 2 wherein the conservative amino acid is selected from the group consisting of alanine, serine, leucine, isoleucine, glutamate and valine.
- 5. The oxidation-resistant mutein of claim 4 selected from the group consisting ala.sub.104 ser.sub.125 IL-2, ala.sub.104 ala.sub.125 IL-2, val.sub.104 ser.sub.125 IL-2, val.sub.104 IL-2, val.sub.104 ala.sub.125 IL-2, des-ala.sub.1 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 ala.sub.104 IL-2, des-ala.sub.1 ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 val.sub.104 ser.sub.125 IL-2, des-ala.sub.1 val.sub.104 IL.sub.2, des-ala.sub.1 val.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 ala.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 val.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 val.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 val.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 ala.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 val.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 val.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 val.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 ala.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 val.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 val.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 val.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 ala.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 val.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 val.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 val.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 des-ser.sub.6 ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 des-ser.sub.6 ala.sub.104 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 des-ser.sub.6 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 des-ser.sub.6 ala.sub.104 ser.sub.125 IL-2, des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 des-ser.sub.6 ala.sub.104 IL-2, and des-ala.sub.1 des-pro.sub.2 des-thr.sub.3 des-ser.sub.4 des-ser.sub.5 des-ser.sub.6 ala.sub.104 ser.sub.125 IL-2.
- 6. The oxidation-resistant mutein of claim 5 selected from the group consisting of ala.sub.104 ser.sub.125 IL-2, ala.sub.104 IL.sub.2, ala.sub.104 ala.sub.125 IL-2, des-ala.sub.1 ala.sub.104 IL-2, des-ala.sub.1 ala.sub.104 ser.sub.125 IL-2, or des-ala.sub.1 ala.sub.104 ala.sub.125 IL-2.
- 7. A human IL-2 oxidation-resistant mutein having a conservative amino acid substitution at position 104, wherein the reference IL-2 protein contains a methionine at position 104, and wherein said mutein exhibits the biological activity of human IL-2.
- 8. The mutein of claim 7 wherein the conservative amino acid is selected from the group consisting of glycine, alanine, serine, threonine, valine, isoleucine, leucine, asparagine, glutamine, glutamate, tyrosine, and phenylalanine.
- 9. The mutein of claim 8 wherein the amino acid is selected from the group consisting of alanine, serine, leucine, glutamate, and valine.
- 10. The mutein of claim 9 wherein the protein is unglycosylated and the amino acid is serine or alanine.
- 11. A therapeutic formulation comprising an effective amount of the mutein of claim 7 and an inert, non-allergenic, pharmaceutically compatible carrier.
- 12. The formulation of claim 11 wherein the mutein is an ala.sub.104 IL-2 mutein, and the carrier is selected from the group consisting of distilled water, physiological saline, Ringer's solution, and Hank's solution.
- 13. The formulation of claim 12, further comprising non-toxic stabilizers and solubilizers.
Parent Case Info
This is a division of Ser. No. 893,186, filed Aug. 5, 1986, now U.S. Pat. No. 4,752,585 which is a continuation-in-part application of copending U.S. application Ser. No. 810,656 filed Dec. 17, 1985 now abandoned which is a continuation-in-part application of copending U.S. application Ser. No. 692,596 filed Jan. 18, 1985 now abandoned.
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Number |
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8700817 |
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WOX |
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Divisions (1)
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Number |
Date |
Country |
Parent |
893186 |
Aug 1986 |
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Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
810656 |
Dec 1985 |
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Parent |
692596 |
Jan 1985 |
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