G-CSF conjugates
Physiologically active PEG-GCSF conjugates having a formula as follows: The manual explains how to pick up Headings and Sideheadings Use “uppercase” when are described, as well as compositions containing a mixture of each conjugates in which m and n can be different integers for the conjugates in the composition.
1 . A physiologically active conjugate having the formula
wherein G is a granulocyte colony stimulating factor less the amino group or groups thereof which form an amide linkage with a polyethylene glycol moiety in the conjugate; R is lower alkyl; n is an integer from 420 to 550; and m is an integer from 1 to 5.
2 . The conjugate of claim 1 wherein R is methyl.
3 . The conjugate of claim 2 wherein n is from 450 to 490.
4 . The conjugate of claim 2 wherein m is an integer from 1 to 4.
5 . The conjugate of claim 4 wherein m is 2.
6 . The conjugate of claim 1 wherein the granulocyte colony stimulating factor is GCSF Mutein having the sequence shown in FIG. 1 .
7 . The conjugate of claim 1 wherein n is from 450 to 490.
8 . The conjugate of claim 1 wherein m is an integer from 1 to 4.
9 . The conjugate of claim 8 wherein m is 2.
10 . The conjugate of claim 1 wherein the granulocyte colony stimulating factor is GCSF Mutein having the sequence shown in FIG. 1 .
11 . The conjugate of claim 1 , which has a longer circulating half-life and greater in vivo granulopoietic activity than the corresponding unconjugated granulocyte colony stimulating factor.
12 . The conjugate of claim 11 , wherein the granulocyte colony stimulating factor is GCSF Mutein having the sequence shown in FIG. 1 .
13 . A physiologically active conjugate having the formula
wherein R is methyl; n is an integer from 450 to 490; m is 2; and G is GCSF Mutein having the sequence shown in FIG. 1 less the amino groups thereof which form an amide linkage with a polyethylene glycol moiety in the conjugate.
14 . A composition comprising physiologically active conjugates having the formula
wherein G in each of the conjugates is a granulocyte colony stimulating factor less the amino group or groups thereof which form an amide linkage with a polyethylene glycol moiety in the conjugates; R in each of the conjugates is independently lower alkyl; n in each of the conjugates is independently an integer from 420 to 550; in each of the conjugates m is independently an integer from 1 to 4; the percentage of conjugates where m is 1 is from eighteen to twenty-five percent; the percentage of conjugates where m is 2 is from fifty to sixty-six percent; the percentage of conjugates where m is 3 is from twelve to sixteen percent; and the percentage of conjugates where m is 4 is up to five percent.
15 . The composition of claim 14 , wherein R is methyl in each of the conjugates.
16 . The composition of claim 14 , wherein n and R are the same in each of the conjugates.
17 . The composition of claim 14 wherein n is from 450 to 490.
18 . The composition of claim 14 where in each of the conjugates the granulocyte colony stimulating factor is GCSF Mutein having the sequence shown in FIG. 1 .
19 . A composition comprising physiologically active conjugates having the formula
wherein R is methyl; n in each of the conjugates is the same and is an integer from 450 to 490; G is GCSF Mutein having the sequence shown in FIG. 1 less the amino groups thereof which form an amide linkage with a polyethylene glycol moiety in the conjugate; in each of the conjugates m is independently an integer from 1 to 4; the percentage of conjugates where m is 1 is from eighteen to twenty-five percent; the percentage of conjugates where m is 2 is from fifty to sixty-six percent; the percentage of conjugates where m is 3 is from twelve to sixteen percent; and the percentage of conjugates where m is 4 is up to five percent.
20 . A method for producing a PEG-GCSF conjugate having a longer circulating half-life and greater in vivo granulopoietic activity than the corresponding unconjugated GCSF, which method consists of covalently linking a reagent of the formula
to the GCSF to produce said conjugate.
21 . The method of claim 21 , wherein the GCSF is GCSF Mutein having the sequence shown in FIG. 1 .