Synthetic bioconjugates
Provided herein are bioconjugates comprising a backbone and at least one branched or unbranched peptide having at least one collagen-binding unit covalently bonded thereto via a spacer and methods of use thereof.
1. A bioconjugate comprising a glycan and from 1 to about 50 binding unit(s) of formula (I) covalently bonded thereto:
(( X 1 ) m X 2 ) n X 3 - L (I)
wherein:
X 1 is an amino acid sequence comprising a collagen-binding unit;
X 2 and X 3 are independently absent, are an amino acid sequence having from 1 to 15 amino acids, or area moiety
where p and q are each independently an integer from 1 to 10;
L is a spacer of from 7 to 20 amino acids selected from the group consisting of glycine (G), serine (S), arginine (R), and lysine (K), provided L comprises at least two arginines (R) within the first five amino acids from the glycan, and wherein L further comprises an optional linking moiety which covalently bonds to the glycan;
m is 1 or 2; and
n is 1 or 2.
2. The bioconjugate of claim 1 , wherein:
X 2 is an amino acid sequence comprising GSG; and
X 3 is an amino acid sequence comprising XRR, where X is absent or is a natural or non-natural amino acid having a side chain capable of forming an amide bond.
3. The bioconjugate of claim 1 , wherein the binding unit of formula (I) is X 1 -GSGSGSRR— (SEQ ID NO: 4).
4. The bioconjugate of claim 1 , wherein X 1 comprises from 3 to about 10 amino acids.
5. The bioconjugate of claim 1 , wherein X 1 comprises the amino acid sequence RRANAALKAGELYKSILY (SEQ ID NO: 9).
6. The bioconjugate of claim 1 , wherein X 1 comprises the amino acid sequence RRANAALKAGQLYKSILY (SEQ ID NO: 13).
7. The bioconjugate of claim 1 , wherein X 2 comprises from 3 to 5 amino acids and X 3 comprises from 2 to 4 amino acids.
8. The bioconjugate of claim 1 , wherein the glycan comprises from about 1 to about 50% functionalization, or from about 5 to about 30% functionalization, or about 25% functionalization, wherein the percent (%) functionalization is determined by a percent of disaccharide units on the glycan which are functionalized with the binding unit of formula (I).
9. The bioconjugate of claim 1 , wherein the glycan is heparin or a derivative thereof.
10. The bioconjugate of claim 1 , wherein the bioconjugate comprises about 8 or about 5 binding unit(s) of formula (I) covalently bonded thereto.
11. The bioconjugate of claim 1 , wherein the binding unit is RRANAALKAGELYKSILYGSGSGSRR—NHNH— (SEQ ID NO: 128).
12. The bioconjugate of claim 1 , wherein the binding unit is RRANAALKAGQLYKSILYGSGSGSRR—NHNH— (SEQ ID NO: 129).
13. The bioconjugate of claim 1 , wherein the binding unit is GQLYKSILYGSGSGSRR—NHNH— (SEQ ID NO: 6).
14. The bioconjugate of claim 1 , wherein the binding unit is (GQLYKSILYGSG) 2 -KSGSRR—NHNH— (SEQ ID NO: 59 and SEQ ID NO: 130).
15. A bioconjugate comprising heparin conjugated to GQLYKSILYGSGRRGSG (SEQ ID NO: 95) in a 1:8 ratio or in a 1:5 ratio.
16. A binding unit of formula (II):
(( X 1 ) m X 2 ) n X 3 - L (II)
wherein:
X 1 is an amino acid sequence comprising a collagen-binding unit comprising from 3 to about 10 amino acids;
X 2 and X 3 are independently absent, are an amino acid sequence having from 1 to 15 amino acids, or area moiety
where p and q are each independently an integer from 1 to 10;
L is a spacer of from 7 to 20 amino acids selected from the group consisting of glycine (G), serine (S), arginine (R), and lysine (K), provided L comprises at least two arginines (R) within the first five amino acids from the terminus, and wherein L further comprises an optional linking moiety;
m is 1 or 2; and
n is 1 or 2;
provided that the binding unit is not RRRKKIQGRSKR (SEQ ID NO: 2) or RRGGRKWGSFEG (SEQ ID NO: 3).
17. The binding unit of claim 16 , wherein:
X 2 is an amino acid sequence comprising GSG;
X 3 is an amino acid sequence comprising RR, KRR, GSG, or KGSG (SEQ ID NO: 73).
18. The binding unit of claim 16 , wherein X 1 comprises the amino acid sequence GQLYKSILY (SEQ ID NO: 5) or the amino acid sequence RRANAALKAGELYKSILY (SEQ ID NO: 9).
19. A peptide having the amino acid sequence RRANAALKAGELYKSILYGSGSGSRR (SEQ ID NO: 128), RRANAALKAGELYKSILYGSGSGSRR—NHNH 2 (SEQ ID NO: 128), RRANAALKAGQLYKSILYGSGSGSRR (SEQ ID NO: 6), RRANAALKAGQLYKSILYGSGSGSRR—NHNH 2 (SEQ ID NO: 6), (GQLYKSILYGSG) 2 -KSGSRR (SEQ ID NO: 59 and SEQ ID NO: 130), or (GQLYKSILYGSG) 2 -KSGSRR—NHNH 2 (SEQ ID NO: 59 and SEQ ID NO: 130).
20. A method for treating peripheral artery disease in a patient in need thereof, comprising administering an effective amount of a bioconjugate of claim 19 .
21. A method for treating fibrosis in a patient in need thereof, comprising administering an effective amount of a bioconjugate of claim 19 .
22. The bioconjugate of claim 19 , wherein L is selected from the group consisting of GSGSGRR (SEQ ID NO: 112), GSGSGSRR (SEQ ID NO: 4), GSGSGSGRR (SEQ ID NO: 113), GSGSGSGSRR (SEQ ID NO: 114), GSGSGSGSGSGRR (SEQ ID NO: 115), GSGSGSGSGSGSRR (SEQ ID NO: 116), GSGSGSGSGSGSGRR (SEQ ID NO: 117), GSGRRGSG (SEQ ID NO: 119), GSGRRRGSG (SEQ ID NO: 120), GSGRRRR (SEQ ID NO: 122), GSGSGSRRR (SEQ ID NO: 123), GSGSGSRRRR (SEQ ID NO: 124), GSGSGSRRRRR (SEQ ID NO: 125), GSGSGSRRRRRR (SEQ ID NO: 126), and GSGSGSGSRRR (SEQ ID NO: 127).