Nucleic acids and protein variants of hG-CSF with granulopoietic activity
The invention relates to novel granulopoietic activity (GPA) proteins and nucleic acids. The invention further relates to the use of the GPA proteins in the treatment of G-CSF related disorders.
1. A variant hG-CSF protein comprising amino acid substitutions at positions 17 and 28 as compared to the protein of SEQ ID NO:2, wherein said HG-CSF protein has increased thermal stability as compared to the protein of SEQ ID NO: 2.
2. A variant hG-CSF orotein according to claim 1 wherein said hG-CSF protein comprises at least one further substitution at a cosition selected from the arouo consisting of positions 14, 20, 21, 24, 27, 31, 32, 34, 35, 38, 78, 79, 82, 85, 89, 91, 92, 99, 102, 103, 108, 107, 109, 110, 113, 116, 117, 120, 145, 148, 147, 148, 151, 153, 155, 156, 157, 160, 161, 164, 167, 168 and 170.
3. A variant hG-CSF protein according to claim 1 , wherein said hG-CSF protein further comprises at least one of further substitution at a position selected from the group consisting of the positions 78, 85, 89, 92, 103, 110, 113, 151, 153, and 168.
4. A variant hG-CSF protein according to claim 3 , wherein said substitutions are 17A and 28A, and said further substitution is 110I (SEQ ID NO: 19).
5. A variant hG-CSF orotein according to claim 3 , wherein said substitutions are 17L and 28A, and said further substitutions are 78F, 85F, 151I, 153I, and 168F (SEQ ID NO: 20).
6. A variant hG-CSF protein according to claim 3 , wherein said substitutions are 17L and 28A, and said further substitutions are 78F, 85F, and 168F (SEQ ID NO: 21).
7. A variant hG-CSF protein according to claim 3 , wherein said substitutions are 17A and 28A, and said further substitutions are 89F, 92F, 103V, and 110I (SEQ ID NO: 22).
8. A variant hG-CSF protein according to claim 3 , wherein said substitutions are 17A and 28A, and said further substitutions are 89F, and 92F (SEQ ID NO: 23).
9. A variant hG-CSF protein according to claim 3 , wherein said substitutions are 17A and 28A (SEQ ID NO: 24).
10. A variant hG-CSF protein according to claim 3 , wherein said substitutions are 17L and 28A, and said further substitutions are 78F, 85F, 113L, 151I and 153I (SEQ ID NO: 25).
11. A recombinant nucleic acid encoding the an hG-CSF Drotein of claim 1 .
12. A pharmaceutical composition comprising an hG-CSF protein according to claim 1 and a pharmaceutically acceptable carrier.