NEW CONJUGATES OF PEPTIDES AND POLYSACCHARIDE

Abstract

Provided is a conjugate formed between one or more linear polysaccharide chains and a peptide selected from one or more of the peptide components (a), (b), (c) or (d) as defined below: (a) a peptide component of formula (I), A-Q-B I wherein Q represents a structural fragment of formula (II), wherein: the squiggly lines and m have meanings given in the description and A and B have meanings given in the description, but may represent a peptide component of the amino acid sequence: [W-Lys-X1-Ser-U-X2-Y] n-W-Lys-X1-Ser-U-X2-Y--- (SEQ ID No: 3), wherein the dashed line, n, W, X1, U, X2 and Y have meanings given in the description; (b) a peptide component of the amino acid sequence: [Ala-Lys-X1-Ser-U-X2-Y] p-Ala-Lys-X1-Ser-U-X1-Y-G (SEQ ID No: 4) wherein p, G, X1, U, X2 and Y have meanings given in the description; (c) a peptide component of the amino acid sequence: W-Lys-X1-Ser-U-X2-Y-G (SEQ ID No: 5), wherein W, X1, U, X2, Y and G have meanings given in the description; or (d) a peptide component of the amino acid sequence: K-W1-Lys-X1-Ser-U1-X2-Y1-I-J (SEQ ID No: 6), wherein K, W1, U1, Y1, I, J, X1 and X2 have meanings as well as regioisomers, stereoisomers, and pharmaceutically- or cosmetically-acceptable salts of said conjugates, which conjugates are particularly useful in the treatment of conditions characterised by inflammation, including wounds, burns, and disorders of the mucosa, such as anorectal diseases, inflammatory bowel diseases, gynaecological diseases and dental diseases. Preferred linear polysaccharides include hyaluronic acid.

##STR00001##

Claims

1. A conjugate formed between one or more linear polysaccharide chains and a peptide, which peptide component is selected from one or more of the peptide components (a), (b), (c) or (d) as defined below: (a) a peptide component of formula I,
A-Q-B  I wherein: A and B independently represent Z or A.sup.1-Q.sup.1-B.sup.1; Q represents a structural fragment of formula II, ##STR00005## wherein: the squiggly lines represent points of attachment of Q to A and/or B; and m represents an integer 1 to 4; A.sup.1 and B.sup.1 independently represent Z or A.sup.2-Q.sup.2-B.sup.2; A.sup.2 and B.sup.2 independently represent Z or Z-Q.sup.3-Z; Q.sup.1, Q.sup.2 and Q.sup.3 independently represent structural fragments of formula III, ##STR00006## wherein: the squiggly lines adjacent to the NH groups represent the points of attachment of Q.sup.1, Q.sup.2 and Q.sup.3 to A.sup.1 and/or B.sup.1, A.sup.2 and/or B.sup.2, and Z, respectively; and the squiggly line adjacent to the O atom represents the point of attachment of Q.sup.1, Q.sup.2 and Q.sup.3 to Q, Q.sup.1 and Q.sup.2, respectively; and m is as defined above; on each occasion that it is employed, Z represents a peptide component of the amino acid sequence: TABLE-US-00028 (SEQ ID No: 3) [W-Lys-X.sup.1-Ser-U-X.sup.2-Y].sub.n-W-Lys-X.sup.1-Ser-U-X.sup.2-Y--- wherein: the dashed line represents the point of attachment of Z to the rest of the molecule; n represents 0 or an integer 1 to 4; and, on each occasion that they are employed: W is absent or represents a 1 or 2 amino acid sequence, in which the amino acids are selected from one or more of the group Lys, Ala, DOPA and a 3,4-dihydrocinnamic acid (HCA) residue, provided that, when present, the HCA residue is located at the N-terminus of the peptide sequence Z; X.sup.1 represents Pro, Hyp or diHyp; U represents Tyr, DOPA or a single bond; X.sup.2 represents Ser, Pro, Hyp or diHyp; Y represents a single bond or represents a 1 to 5 amino acid sequence, in which the amino acids are selected from one or more of the group Lys, Ala, Pro, Hyp, diHyp, Thr, DOPA and Tyr; or (b) a peptide component of the amino acid sequence: TABLE-US-00029 (SEQ ID No: 4) [Ala-Lys-X.sup.1-Ser-U-X.sup.2-Y].sub.p-Ala-Lys-X.sup.1-Ser-U-X.sup.2-Y-G wherein: p represents an integer 1 to 4; G is absent or represents DOPA or dopamine; and X.sup.1, U, X.sup.2 and Y are as defined above; or (c) a peptide component of the amino acid sequence: TABLE-US-00030 (SEQ ID No: 5) W-Lys-X.sup.1-Ser-U-X.sup.2-Y-G wherein W, X.sup.1, U, X.sup.2, Y and G are as defined above; or (d) a peptide component of the amino acid sequence: TABLE-US-00031 (SEQ ID No: 6) K-W.sup.1-Lys-X.sup.1-Ser-U.sup.1-X.sup.2-Y.sup.1-I-J wherein K represents an optional N-terminal HCA group; W may be absent or represents a 1 or 2 amino acid sequence, in which the amino acids are selected from one or more of the group Ser, Lys, Ala and DOPA; U.sup.1 represents Tyr, DOPA or a single bond; Y.sup.1 represents a single bond or represents Y; I represents Pro, Hyp, diHyp, Thr, DOPA or Tyr; and J represents Lys or is absent; and X.sup.1, X.sup.2 and Y are as defined above, as well as regioisomers, stereoisomers, and pharmaceutically- or cosmetically-acceptable salts of said conjugate.

2. The conjugate as claimed in claim 1, wherein the polysaccharide chain is hyaluronic acid.

3. The conjugate as claimed in claim 1, wherein X.sup.1 represents Pro.

4. The conjugate as claimed in claim 1, X.sup.2 represents Pro or Hyp.

5. The conjugate as claimed in claim 1, wherein W represents HCA, HCA-Ala-, Ala, DOPA, Lys-Ala or DOPA-Ala-.

6. (canceled)

7. The conjugate as claimed in claim 1, wherein Y represents a 4 amino acid sequence, in which the amino acids are selected from one or more of the group Lys, Ala, Hyp, Thr, DOPA and Tyr.

8. The conjugate as claimed in claim 7, wherein Y represents an amino acid sequence selected from the group -Hyp-Y.sup.1-Y.sup.2-Lys- (SEQ ID No: 156) and -Thr-Y.sup.1-Y.sup.2-Lys- (SEQ ID No: 157), wherein Y.sup.1 and Y.sup.2 are each independently selected from the group Ala, Hyp, Thr, DOPA and Tyr.

9. The conjugate as claimed in claim 7, wherein the amino acid sequence defined by Y is selected from the group -Pro-Thr-DOPA-Lys- (SEQ ID No: 158), -Pro-Thr-Tyr-Lys- (SEQ ID No: 159), -Thr-Tyr-Pro-Lys- (SEQ ID No: 160), -Thr-DOPA-Pro-Lys- (SEQ ID No: 161), -Hyp-Thr-Tyr-Lys- (SEQ ID No: 162), -Hyp-Thr-DOPA-Lys- (SEQ ID No: 163), -Hyp-Thr-Ala-Lys- (SEQ ID No: 164), -Thr-Tyr-Hyp-Lys- (SEQ ID No: 165), -Thr-DOPA-Hyp-Lys- (SEQ ID No: 166), -Thr-Ala-Hyp-Lys- (SEQ ID No: 167), -Hyp-Thr-, -Thr-Tyr-, -Pro-Thr-, -Thr-DOPA-, -Thr-Tyr-Lys-, -Tyr-Pro-Lys-, -DOPA-Pro-Lys, -Hyp-Thr-Tyr-, -Thr-Tyr-Hyp-Lys-DOPA- (SEQ ID No: 169), -Hyp-Thr-Tyr-Hyp-Lys- (SEQ ID No: 168) and -Hyp-Thr-DOPA.

10. The conjugate as claimed in claim 1, wherein m represents 4.

11. The conjugate as claimed in claim 1, wherein A and B both represent Z, or both represent A.sup.1-Q.sup.1-B.sup.1.

12. The conjugate as claimed in claim 1, wherein A.sup.1 and B.sup.1 both represent Z, or both represent A.sup.2-Q.sup.2-B.sup.2.

13. The conjugate as claimed in claim 1, wherein A.sup.2 and B.sup.2 both represent Z, or both represent Z-Q.sup.3-Z.

14. The conjugate as claimed in claim 1, wherein n is 0.

15. (canceled)

16. The conjugate as claimed in claim 1, wherein G is absent and the peptide component as defined under (b) comprises the amino acid sequence: TABLE-US-00032 (SEQ ID No: 7) Ala-Lys-Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys-Ala-Lys- Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys; (SEQ ID No: 8) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys- Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser- Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp- Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr- Tyr-Lys; (SEQ ID No: 9) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 10) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys- Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro- Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser- DOPA-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 11) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 12) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys- Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 13) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys- Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro- Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys; or (SEQ ID No: 14) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys- Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro- Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser- Tyr-Hyp-Hyp-Thr-DOPA-Lys.

17. The conjugate as claimed in claim 1, wherein U represents Tyr and/or W represents Ala-.

18. The conjugate as claimed in claim 17, wherein Z in the peptide component defined under (a) is selected from the group: TABLE-US-00033 (SEQ ID No: 2) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 1) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 15) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 16) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 17) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys-DOPA---; (SEQ ID No: 18) Ala-Lys-Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys-Ala-Lys- Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys--;  and (SEQ ID No: 19) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys- Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser- Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp- Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr- Tyr-Lys---.

19. The conjugate as claimed in claim 1, wherein U represents Tyr and/or W represents Lys-Ala-.

20. The conjugate as claimed in claim 19, wherein Z in the peptide component defined under (a) is selected from the group: Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr--- (SEQ ID No: 37); Lys-Ala-Lys-Hyp-Ser-Tyr-Hyp-Hyp-Thr-DOPA--- (SEQ ID No: 38); and Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA--- (SEQ ID No: 39).

21. The conjugate as claimed in claim 1, wherein U represents Tyr and/or W represents HCA-, HCA-Ala-, DOPA- or DOPA-Ala-.

22. The conjugate as claimed in claim 21, wherein Z in the peptide component defined under (a) is selected from the group: TABLE-US-00034 (SEQ ID No: 45) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 46) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 47) HCA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys---; (SEQ ID No: 48) HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---; (SEQ ID No: 49) DOPA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---; (SEQ ID No: 50) DOPA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys---; (SEQ ID No: 51) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---;  and (SEQ ID No: 52) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---.

23. The conjugate as claimed in claim 1, wherein U represents DOPA and/or W represents Ala or Lys-Ala-.

24. The conjugate as claimed in claim 23, wherein Z in the peptide component defined under (a) is selected from the group: TABLE-US-00035 (SEQ ID No: 65) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 66) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 67) Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA---; (SEQ ID No: 68) Lys-Ala-Lys-Hyp-Ser-DOPA-Hyp-Hyp-Thr-DOPA---; (SEQ ID No: 69) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala- lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---;  and (SEQ ID No: 70) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala- Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys- Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro- Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser- DOPA-Hyp-Hyp-Thr-DOPA-Lys---.

25. The conjugate as claimed in claim 1, wherein U represents DOPA and/or W represents HCA-, HCA-Ala-, DOPA- or DOPA-Ala-.

26. The conjugate as claimed in claim 25, wherein Z is selected from the group: TABLE-US-00036 (SEQ ID No: 94) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 95) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 96) HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---; (SEQ ID No: 96) HCA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---; (SEQ ID No: 98) DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---; (SEQ ID No: 99) DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---; (SEQ ID No: 100) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 101) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys---; and (SEQ ID No: 102) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---.

27. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (a), A and B both represent Z, and one or both Z groups represent: TABLE-US-00037 (SEQ ID No: 1) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 2) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 15) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 16) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 45) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 48) HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---; (SEQ ID No: 51) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 65) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 67) Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA---; (SEQ ID No: 94) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 99) DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---; or (SEQ ID No: 100) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---, and Q represents a Lys residue.

28. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (a), A and B both represent A.sup.1-Q.sup.1-B.sup.1, A.sup.1 and B.sup.1 both represent Z, and one or both Z groups represent: TABLE-US-00038 (SEQ ID No: 1) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 2) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 15) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 16) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 38) ;Lys-Ala-Lys-Hyp-Ser-Tyr-Hyp-Hyp-Thr-DOPA---; (SEQ ID No: 39) Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA---; (SEQ ID No: 45) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 46) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 51) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 65) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 66) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 96) HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---; (SEQ ID No: 98) DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---; or (SEQ ID No: 100 DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---, and Q.sup.1 represents a Lys residue.

29. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (a), A and B both represent A.sup.1-Q.sup.1-B.sup.1, A.sup.1 and B.sup.1 both represent A.sup.2-Q.sup.2-B.sup.2, A.sup.2 and B.sup.2 both represent Z, one or both Z groups represent: TABLE-US-00039 (SEQ ID No: 1) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---; (SEQ ID No: 2) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 15) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 16) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---; (SEQ ID No: 45) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 51) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---; (SEQ ID No: 52) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---; (SEQ ID No: 66) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---; or (SEQ ID No: 100) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---, and Q.sup.1 and Q.sup.2 both represent Lys residues.

30. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (a), A and B both represent A.sup.1-Q.sup.1-B.sup.1, A.sup.1 and B.sup.1 both represent A.sup.2-Q.sup.2-B.sup.2, A.sup.2 and B.sup.2 both represent Z-Q.sup.3-Z, one or both Z groups represent: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys--- (SEQ ID No: 1); or Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 2), and Q.sup.1, Q.sup.2 and Q.sup.3 all represent Lys residues.

31. The conjugate as claimed in claim 17, wherein the peptide component defined under (c) comprises the amino acid sequence: TABLE-US-00040 (SEQ ID No: 20) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys-DOPA; (SEQ ID No: 21) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys-Dopamine; (SEQ ID No: 22) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Dopamine; (SEQ ID No: 23) Ala-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-DOPA-Lys; (SEQ ID No: 24) Ala-Lys-Pro-Ser-Tyr-Pro-Thr-Tyr-Pro-Lys; (SEQ ID No: 25) Ala-Lys-Pro-Ser-Tyr-Pro-Thr-DOPA-Pro-Lys; (SEQ ID No: 26) Ala-Lys-Pro-Ser-Tyr-Pro-Hyp-Thr-Tyr-Lys; (SEQ ID No: 27) Ala-Lys-Pro-Ser-Tyr-Pro-Hyp-Thr-DOPA-Lys; (SEQ ID No: 28) Ala-Lys-Pro-Ser-Tyr-Hyp-Pro-Thr-Tyr-Lys; (SEQ ID No: 29) Ala-Lys-Pro-Ser-Tyr-Hyp-Pro-Thr-DOPA-Lys; (SEQ ID No: 30) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 31) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 32) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-DOPA; (SEQ ID No: 33) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-DOPA; (SEQ ID No: 34) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Dopamine; (SEQ ID No: 35) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys-DOPA; or (SEQ ID No: 36) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys-Dopamine.

32. The conjugate as claimed in claim 19, wherein the peptide component defined under (c) comprises the amino acid sequence: TABLE-US-00041 (SEQ ID No: 40) Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA; (SEQ ID No: 41) Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Dopamine; and (SEQ ID No: 42) Lys-Ala-Lys-Hyp-Ser-Tyr-Hyp-Hyp-Thr-DOPA; (SEQ ID No: 43) Lys-Ala-Lys-Hyp-Ser-Tyr-Hyp-Hyp-Thr-Tyr; or (SEQ ID No: 44) Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr.

33. The conjugate as claimed in claim 21, wherein the peptide component defined under (c) comprises the amino acid sequence: TABLE-US-00042 (SEQ ID No: 53) DOPA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys; (SEQ ID No: 54) DOPA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys; (SEQ ID No: 55) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 56) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys; (SEQ ID No: 57) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 58) DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 59) HCA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys; (SEQ ID No: 60) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 61) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 62) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys; (SEQ ID No: 63) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys; or (SEQ ID No: 64) HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys.

34. The conjugate as claimed in claim 23, wherein the peptide component defined under (c) comprises the amino acid sequence: TABLE-US-00043 (SEQ ID No: 71) Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Dopamine; (SEQ ID No: 72) Lys-Ala-Lys-Hyp-Ser-DOPA-Hyp-Hyp-Thr-DOPA; (SEQ ID No: 73) Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA; (SEQ ID No: 74) Ala-Lys-Pro-Ser-DOPA-Pro-Pro-Thr-Tyr-Lys; (SEQ ID No: 75) Ala-Lys-Pro-Ser-DOPA-Pro-Pro-Thr-DOPA-Lys; (SEQ ID No: 76) Ala-Lys-Pro-Ser-DOPA-Pro-Thr-Tyr-Pro-Lys; (SEQ ID No: 77) Ala-Lys-Pro-Ser-DOPA-Pro-Thr-DOPA-Pro-Lys; (SEQ ID No: 78) Ala-Lys-Pro-Ser-DOPA-Pro-Hyp-Thr-Tyr-Lys; (SEQ ID No: 79) Ala-Lys-Pro-Ser-DOPA-Pro-Hyp-Thr-DOPA-Lys; (SEQ ID No: 80) Ala-Lys-Pro-Ser-DOPA-Hyp-Pro-Thr-Tyr-Lys; (SEQ ID No: 81) Ala-Lys-Pro-Ser-DOPA-Hyp-Pro-Thr-DOPA-Lys; (SEQ ID No: 82) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 83) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 84) Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr; (SEQ ID No: 85) Lys-Ala-Lys-Hyp-Ser-DOPA-Hyp-Hyp-Thr-Tyr; (SEQ ID No: 86) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys-DOPA; (SEQ ID No: 87) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-DOPA; (SEQ ID No: 88) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys-Dopamine; (SEQ ID No: 89) Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Dopamine; (SEQ ID No: 90) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-Tyr-Hyp-Lys-DOPA; (SEQ ID No: 91) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys-DOPA; (SEQ ID No: 92) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-Tyr-Hyp-Lys-Dopamine; or (SEQ ID No: 93) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys-Dopamine.

35. The conjugate as claimed in claim 25, wherein the peptide component defined under (c) comprises the amino acid sequence: TABLE-US-00044 (SEQ ID No: 103) DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys; (SEQ ID No: 104) DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys; (SEQ ID No: 105) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 106) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys; (SEQ ID No: 107) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 108) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 109) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 110) HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys; (SEQ ID No: 111) HCA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys; (SEQ ID No: 112) DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 113) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-Tyr-Hyp-Lys; or (SEQ ID No: 114) HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys.

36. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), W.sup.1 represents Ala or Ser, or is absent.

37. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), X.sup.2 represents Pro, Hyp or diHyp.

38. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), when K is not present, W.sup.1 represents Ala or is absent and J represents Lys, then I represents Pro, Hyp, diHyp or Thr.

39. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), in the sequence defined by Y.sup.1: the amino acid DOPA, Thr or Tyr is linked to I; and/or the amino acid Pro, Hyp or Thr is linked to X.sup.2.

40. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), K is absent.

41. The conjugate as claimed in claim 40, wherein the peptide component defined under (d) comprises the amino acid sequence: TABLE-US-00045 (SEQ ID No: 115) Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 116) Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 117) Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 118) Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 119) Ser-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys; (SEQ ID No: 120) Ser-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys; (SEQ ID No: 121) Ser-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 122) Ser-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 123) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 124) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 125) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys; or (SEQ ID No: 126) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys.

42. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), J is absent.

43. The conjugate as claimed in claim 42, wherein the peptide component defined under (d) comprises the amino acid sequence: TABLE-US-00046 (SEQ ID No: 127) Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp; (SEQ ID No: 128) Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 129) Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp; (SEQ ID No: 130) Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 131) Ser-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr; (SEQ ID No: 132) Ser-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp; (SEQ ID No: 133) Ser-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 134) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp; (SEQ ID No: 135) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 136) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp; (SEQ ID No: 137) Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 138) Ser-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA; (SEQ ID No: 146) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr; (SEQ ID No: 147) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 148) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp; (SEQ ID No: 149) Ala-Lys-Pro-Ser-Pro-Thr-Tyr-Pro; (SEQ ID No: 150) Ala-Lys-Hyp-Ser-Tyr-Hyp-Thr-Tyr-Hyp; (SEQ ID No: 151) Ala-Lys-Hyp-Ser-DOPA-Hyp-Thr-DOPA-Hyp; or (SEQ ID No: 152) Ala-Lys-Hyp-Ser-Tyr-Hyp-Thr-DOPA-Hyp.

44. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), K is an N-terminal HCA group.

45. The conjugate as claimed in claim 44, wherein the peptide component defined under (d) comprises the amino acid sequence: TABLE-US-00047 (SEQ ID No: 139) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys; or (SEQ ID No: 140) HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys.

46. The conjugate as claimed in claim 1, wherein, in the peptide component defined under (d), W.sup.1 is Ala and J is Lys.

47. The conjugate as claimed in claim 46, wherein the peptide component defined under (d) comprises the amino acid sequence: TABLE-US-00048 (SEQ ID No: 141) Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys; (SEQ ID No: 142) Ala-Lys-Pro-Ser-Pro-Thr-Tyr-Pro-Lys; (SEQ ID No: 143) Ala-Lys-Hyp-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys; (SEQ ID No: 144) Ala-Lys-Hyp-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys; or (SEQ ID No: 145) Ala-Lys-Hyp-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys.

48-49. (canceled)

50. A pharmaceutical formulation comprising a conjugate as defined in claim 1 and a pharmaceutically- or cosmetically-acceptable, adjuvant, diluent or carrier.

51. The pharmaceutical formulation as claimed in claim 50 that is suitable for, adapted for, and/or packaged and presented for, topical administration, wherein the pharmaceutically- or cosmetically-acceptable adjuvant, diluent or carrier is a topical adjuvant, diluent or carrier.

52. The pharmaceutical formulation as claimed in claim 50, which is in the form of a gel, a spray, a cream, an ointment or a dry powder.

53. The pharmaceutical formulation as claimed in claim 50 that is suitable for, adapted for, and/or packaged and presented for, administration by injection.

54. (canceled)

55. The pharmaceutical formulation as claimed in claim 50, which further includes a pharmaceutically-active ingredient.

56. A kit of parts comprising components: (A) the conjugate as defined in claim 1; and (B) a pharmaceutical formulation including a pharmaceutically-active ingredient in admixture with a pharmaceutically-acceptable adjuvant, diluent or carrier, which components (A) and (B) are each provided in a form that is suitable for administration in conjunction with the other.

57. The pharmaceutical formulation as claimed in claim 55, wherein the pharmaceutically-active ingredient is an anti-inflammatory agent, a pro-inflammatory agent, an antibiotic, an anti-bacterial and/or antiprotozoal agent, an antiviral agent, an anaesthetic and/or a wound recovery drug.

58. The pharmaceutical formulation as claimed in claim 57, wherein the pharmaceutically-active ingredient is an anti-inflammatory agent.

59-60. (canceled)

61. A method of treatment of inflammation, an inflammatory disorder, and/or of a disorder characterised by inflammation, which method comprises the administration of a conjugate as defined in claim 1 to a patient in need of such treatment.

62. The method as claimed in claim 61, wherein the inflammation, inflammatory disorder, and/or disorder characterised by inflammation is selected from the group of radiation vaginitis, fibrosis of the vagina and/or nasosinusitis.

63. The method as claimed in claim 61, wherein the disorder characterised by inflammation is, or results in, a wound or a burn.

64. The method as claimed in claim 63, wherein the disorder resulting in a wound is hemorrhoids or ulcerative colitis.

65. The method as claimed in claim 61, wherein the conjugate(s) or salt thereof is administered topically in the form of a topical formulation.

66. The method as claimed in claim 65 wherein the relevant condition is treated by way of direct topical administration to the skin.

67. The method as claimed in claim 65, wherein the administration is carried out by way of direct topical administration to a mucosal surface.

68. The method as claimed in claim 61, wherein the conjugate(s) is/are administered by oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal, pulmonary or anorectal delivery.

69. (canceled)

70. The method as claimed in claim 61, wherein the conjugate is crosslinked, before or after administration to the patient.

71-77. (canceled)

Description

EXAMPLES

Illustrative Example 1

(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys).SUB.2 .(SEQ ID No: 11)

[0267] Fmoc-Lys(Boc)-Wang resin (9.15 g, GLS180322-41301, GL Biochem, Shanghai, China) was loaded into a glass reaction column.

[0268] Methylene chloride (DCM, 200 mL; Shandong Jinling Chemical Industry Co. Ltd., Shandong, China) was added to the column and allowed to soak the resin for about half an hour. The DCM was then removed by vacuum filtration.

[0269] The resin was washed 3 times with N,N-dimethylformamide (DMF, 200 mL; Shandong Shitaifeng Fertilizer Industry Co. Ltd., Shandong, China).

[0270] A 20% piperidine solution in DMF (200 mL; Shandong Shitaifeng Fertilizer Industry Co. Ltd., Shandong, China) and was added as deprotection solution and reacted for 20 minutes. The solution was then removed by vacuum filtration and the column was washed with DMF six times.

[0271] Fmoc-DOPA(Acetonide)-OH (4.14 g; GLS190219-21003, GL Biochem. Shanghai, China) and 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium tetrafluoroborate (TBTU, 2.89 g; GLS170805-00705. GL Biochem. Shanghai, China) were added to the resin. DMF (150 mL) was added to the reaction column, followed by N,N-diisopropylethylamine (DIPEA, 2.33 g; Suzhou Highfine Biotech Co. Ltd., Jiangsu, China). A Kiaser test was carried out with few of the resin after 30 min reaction. colour reaction was detected in the resin after 30 minutes, indicating the reaction was complete. The solvent was removed by vacuum filtration.

[0272] The above coupling steps were repeated to couple the remaining amino acids in the same amounts (by mols): Fmoc-Thr(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-Tyr(tBu)—OH, Fmoc-Ser(tBu)—OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH and Fmoc-Ala-OH.

[0273] After the Fmoc-Ala-OH was coupled to the resin, the above coupling steps were repeated starting with Fmoc-Lys(Boc)-OH and followed by Fmoc-DOPA(Acetonide)-OH, Fmoc-Thr(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-Tyr(tBu)—OH, Fmoc-Ser(tBu)—OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH and Fmoc-Ala-OH.

[0274] In a separate procedure, after Fmoc-Ala-OH was coupled on the resin, a deprotection step was carried out to remove the Fmoc protection on Dopa. The resin was washed 3 times with DMF (200 mL each time). A 20% piperidine solution in DMF (200 mL) was added as a deprotection solution and reacted for 20 minutes. Then, the resin was washed three times each with the following solvents, DMF (200 mL each time), DCM (200 mL each time) and methanol (200 mL each time; Xilong Scientific Co. Ltd., Guangdong, China). The resin was dried under vacuum for about 2 hours.

[0275] 160.0 mL (i.e. 10 mL per gram of the dried resin) of lysate, which comprised of 95% trifluoroacetic add (TFA), 2.5% water and 2.5% triisopropylsilane (Tis), were added to immerse the resin-bounded peptide-containing compound. After cleavage for about 2 hours, the solid support was removed by filtration and the filtrate was collected under reduced pressure. The filtrate was precipitated with 1600 mL (i.e. 10 mL per ml of the filtrate) of diethyl ether (Xilong Scientific Co., Ltd., Guangdong, China) and the sediment was collected by filtration. The sediment was dried by vacuum for about 2 hours, yielding 7.53 g of the crude title compound.

[0276] The crude product was firstly analyzed as a 1 mg/mL sample in pure water and detected using a Shimadzu LCMS-8050 system. The analysis column was an Agilent ZORBAX Eclipse SB-C18 (4.6×250 mm, 5 μm column; detection: UV at 220 nm; solvent A: 0.1% TFA in MeCN, solvent B: 0.1% TFA in water, with a linear gradient from 5%-90% solvent A concentration in 50 minutes; flow rate 1.0 mL/min; sample volume: 10 μL).

[0277] The target peak was eluted at 11.926 minutes and had the expected molecular weight, with a purity of 60.345%.

MS: m/z 2380.6

[0278] 7.5 g of the crude product was then dissolved in 80 mL of pure water and purified using LC3000 semi-preparation equipment. The preparation column model was a Dubhe-C18 model (Hanbon Sci. &Tech. Co., Ltd., Jiangsu, China) (50*250 mm, 100 Å column; detection: UV at 220 nm). The appropriate gradient for elution was calculated from LCMS detection step (Solvent A: 0.1% TFA in MeCN, solvent B: 0.1% TFA in water, with a linear gradient from 5%-20% solvent A concentration in 30 minutes; flow rate 60.0 mL/min). The fractions were collected and analyzed using a Shimadzu LC-20 HPLC system (column as above, except with a linear gradient from 5%-30% solvent A concentration in 25 minutes).

[0279] Fractions with a purity of 98% were then mixed for an anion exchange step. This was achieved using a LC3000 semi-preparation equipment (preparation column model: Dubhe-C18 model (as above). The fractions were diluted one time with pure water and loaded to the column directly, after that the column was washed with 0.37% of ammonium acetate in pure water for about 20 minutes followed by pure water for another 20 minutes at the flow rate of 60 mL/min, then eluted with the following gradient (Solvent A: 0.1% HAc in MeCN, solvent B: 0.1% HAc in water, with a linear gradient from 5%-20% solvent A concentration in 30 minutes; flow rate 60.0 mL/min). The fractions were collected and analyzed using Shimadzu LC-20 HPLC system (column and conditions as above). Fractions with a purity of 98% were mixed and freeze-dried to give 3.06 g of the purified title compound.

Illustrative Example 2

[0280]

TABLE-US-00023 (SEQ ID No: 153) (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys).sub.2-Lys

[0281] Fmoc-Lys(fmoc)-Wang resin (9.9 g, GLS191010-41303, GL Biochem, Shanghai, China) was loaded into a glass reaction column.

[0282] The method was the same as described in Illustrative Example 1, except that Fmoc-Lys(Boc)-OH was coupled to the resin first followed by Fmoc-Dopa(Acetonide)-OH, Fmoc-Thr(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-Tyr(tBu)—OH, Fmoc-Ser(tBu)—OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH and Fmoc-Ala-OH, and the amounts of the amino adds, TBTU and DIPEA were doubled (by mols) compared to Illustrative Example 1.

MS: m/z 2508.8

[0283] Repeating essentially the same procedure gave a further batch of crude title compound (yield 7.89 g). Analysis showed a target peak that was eluted at 11.376 minutes with the expected molecular weight (MS: m/z 2508.8). The purity was 68.985%.

[0284] 7.8 g of the crude product was then purified as described in Illustrative Example 1 above to give 2.57 g of pure title compound after freeze-drying.

MS: m/z 2508.8

Illustrative Example 3

[0285]

TABLE-US-00024 (SEQ ID No: 154) {(Dopa-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr- Lys)2-Lys]2-Lys}2-Lys

[0286] Fmoc-Lys(fmoc)-Wang resin (9.9 g, GLS191010-41303, GL Biochem, Shanghai, China) was loaded into a glass reaction column.

[0287] The method was the same as described in Illustrative Example 2, except that Fmoc-Lys(Fmoc)-OH was coupled to the resin first followed by Fmoc-Lys(Fmoc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Tyr(tBu)—OH, Fmoc-Thr(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-4-Hyp(tBu)—OH, Fmoc-Tyr(tBu)—OH, Fmoc-Ser(tBu)—OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH, Fmoc-Ala-OH and Fmoc-Dopa(Acetonide)-OH, and the amounts of the amino acids, TBTU and DIPEA were quadrupled (by mols) compared to Illustrative Example 2.

MS: m/z 11671.1

[0288] Repeating essentially the same procedure gave a further batch of crude title compound (yield 28.89 g). Analysis showed a target peak that was eluted at 11.896 minutes with the expected molecular weight (MS: m/z 11671.1). The purity was 29.985%.

[0289] 28.8 g of the crude product was then purified as described in Illustrative Example 1 above to give 5.57 g of pure title compound after freeze-drying.

Illustrative Example 4

[0290]

TABLE-US-00025 (SEQ ID No: 2) Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys

[0291] The title peptide was synthesised using essentially the same procedure as that described in Illustrative Example 1 above, except that the appropriate amino acids were used in the appropriate peptide coupling sequences.

MS: m/z 1183.3

Example 1

(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys).SUB.2.-Lys (SEQ ID No: 153) HA Conjugate

[0292] A 1% HA solution (50 mM of carboxyl groups) was made by dissolving HA-EP2 (1 g, 19072911, Furida Biotech., Shandong, China) in 100 mL of pure water for 8 hours.

[0293] DMTMM (275 mg, Sigma) and the product of Example 2 (22 mg) were dissolved in 20 mL of pure water to make a DMTMM (50 mM) and peptide (2.5 mM of amino groups) solution. 20 mL of the 1% HA solution was added to the 20 mL DMTMM and peptide solution and stirred at room temperature for one day.

[0294] After one day, 0.1 g of the reacted gel was taken and mixed well with 0.4 mL 96% ethanol. The sample was then centrifuged at 15000 rpm for 15 minutes. The supernatant was taken and detected using a Shimadzu LC-20 HPLC system equipped with LC-20AT binary pump, a DGU-20A5 degasser, a SIL-20AC autosampler, a CTO-20AC column oven, and a SPD-M20A photodiode array detector (Shimadzu, Japan).

[0295] The sample was analysed on an Agilent SB-C18 column (5 μm, 4.6*250 mm; Agilent, USA) under the following conditions: [0296] the mobile phase: 0.1% TFA in acetonitrile (solvent A) and 0.1% TFA in water (solvent B); [0297] linear gradient program: 0-25 min, 10%-35% solvent A; [0298] flow rate: 1.0 mL/min; [0299] column temperature: 30° C.; [0300] the PDA detector recorded UV spectra in the range of from 190 nm to 400 nm; [0301] the HPLC chromatogram was monitored at 220 nm.

[0302] The concentration of the product of Example 2 in the supernatant (un-coupled free peptide) was calculated according its standard curve to be 0.052 mg/mL. Thus, about 0.29 mg/mL (=22/40−0.052*5) of peptide was coupled to HA-EP2, which is about 11.6 mg (=0.29*40) peptide per 200 mg (0.2 g=20*1%) HA-EP2.

[0303] Products are isolated by precipitation by dropwise addition of 96% ethanol (100 mL) to the reaction mixture. The white powder is obtained and thoroughly washed with water:ethanol 1:4, ethanol 96% and finally with absolute ethanol. The product is dried under vacuum for 3 days at 38° C.

Example 2

Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No: 2) HA Conjugate

[0304] HA (200 mg, HA-EP2, Bloomage Biotechnology Co. Ltd., Jinan, China) was taken in a test tube, 2 mL of 0.2M NaOH was added to dissolve the HA powder and mixed well.

[0305] The peptide (SEQ ID No: 2) (220 mg, P200921, USUN Pharmaceutical Co. Ltd., Jiangyin, China) was taken into another test tube. 1 mL of 0.4M HCl was used to dissolve the peptide powder, DMTMM (275 mg, Aladdin, Shanghai, China) was then added to dissolve and mixed well.

[0306] The mixture was then transferred to the HA solution's tube and mixed well, the test tube was then covered, heated at 35° C., and let to react for about 16 hours.

[0307] After 16 hours, the solution became a colorless rigid gel. The gel was removed and cut with 20 mesh screen once. The cut gel was then immersed into 100 mL HA buffer (which contained 9.0 g NaCl, 30 mg KH.sub.2PO.sub.4 and 140 mg Na.sub.2HPO.sub.4.12H.sub.2O in a total volume of 1 L pure water) for one hour. The gel swelled after soaking. The HA buffer was removed by filtration and the gel was cut with 200 mesh screen three times to make small and even particles.

[0308] The small particles were then immersed into 100 mL HA buffer for 1 hour to remove the condensation agent (DMTMM) (dialysis-like step), the HA buffer was then removed by filtration. This step was repeated for additional 7 times to exhaustively remove DMTMM. The smaller particles were obtained at a concentration of about 30 mg/g.

[0309] HA (10 mg, HA-EP2, Bloomage Biotechnology Co. Ltd., Jinan, China) was dissolved into 1 mL HA buffer, 1 g of the prepared small particles was then added and mixed well. The final product was obtained at 20 mg/g.

[0310] The obtained product was then filled in the final packing material and underwent moist heat sterilization.

[0311] The obtained product can be used for filling out wrinkles and folds.

Example 3

[0312] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No: 2) HA Conjugate Combined with Celecoxib

[0313] The HA conjugate is obtained as described in Example 2.

[0314] The HA conjugate and celecoxib are mixed according to the mass ratio of 10:1 and stirred to uniform. The obtained product can be used for intra-articular injection to relieve the symptoms of osteoarthritis and relieve pain.

Example 4

[0315] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No: 2) HA Conjugate Combined with Diclofenac Sodium

[0316] The HA conjugate is obtained as described in Example 2.

[0317] The HA conjugate and diclofenac sodium are mixed according to the mass ratio of 5:1 and stirred to uniform. The obtained product can be used for intra-articular injection as an analgesic and/or anti-inflammatory drug.

Example 5

[0318] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No: 2) HA Conjugate Combined with Rifampicin

[0319] The HA conjugate is obtained as described in Example 2.

[0320] The HA conjugate and Rifampicin are mixed according to the mass ratio of 20:1 and stirred to uniform. The obtained product can be used in ophthalmic surgery as an anti-inflammatory, detumescence and/or analgesic.

Example 6

Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No: 2) HA Conjugate Against Bleeding

[0321] A 49-years old women had allergic rhinitis for about 5 years. She was subject to seasonal allergy every spring and autumn. She was prescribed Fluticasone Propionate Nasal Spray to control the symptoms but started to have dry nose since the previous winter. She had nose bleeds after constant usage of fluticasone propionate nasal spray. When she had another allergy attack in Spring, she used Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No: 2) HA conjugate spray (comprising small particles of the conjugate prepared according to Example 2 above (0.5 mg) dissolved in 1 mL HA buffer in addition to the prescribed drug. She used the conjugate spray 4-5 times during the day. The bleeding stopped and the dry nose felt much better. This indicated that the title compound can be used to relief dry nose symptoms.

Example 7

[0322] {[(Dopa-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys).sub.2-Lys].sub.2-Lys}.sub.2-Lys (SEQ ID No. 154) HA gel

[0323] A gel comprising {[(Dopa-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys).sub.2-Lys].sub.2-Lys}.sub.2-Lys (SEQ ID No. 154) was made by mixing an appropriate amount of the peptide component with methyl cellulose (2.2 g; Shandong Guangda Technology Development Co., Ltd., ShanDong, China) and purified water (75.3 g), and stirring until a homogeneous colloidal suspension was formed. Then, 0.5 g of HA powder (HA-EP2, Bloomage Biotechnology Co. Ltd., Jinan, China), glycerin (11 g) and propanediol 11 g (both from Sinopharm Chemical Reagent Co. Ltd.) were added to the methyl cellulose/water mixture, and the resultant mixture was quickly stirred for 5 minutes to obtain the title product.

[0324] The product of Example 7 is referred to hereinafter as ‘HA gel’.

Example 8

Mouse Ear Swelling Model

[0325] 20 healthy male BALB/c mice of 6-8 weeks of age and average body weight of 18-25 g were supplied by Changzhou Cvens Experimental Animal Co., Ltd. and housed and cared for about for 1 week prior to the experiment. The housing temperature was 25-27° C. with 74% humidity, alternating 12-hour periods of light and darkness, and free access to food and water. The mice were randomly divided into 4 groups as described in Table 1 below, with 5 mice in each group.

[0326] The left ear of each mouse was used as autologous control. The right ear of each mouse was treated by various different treatments, as summarized in Table 1 below. 20 μL of xylene (Shanghai Aladdin Bio-Chem Technology Co., Ltd., Shanghai, China) was applied to the right ear of each mouse, both inside and outside. The ear started to swell in about 4 minutes. Then, 40 μL of treatments or vehicle were applied to the right ears in each group. The mice were put back into their cages.

[0327] Dexamethasone Acetate Cream (DEX, Tianjin Jinyao Pharmaceutical Co., Ltd.) was used as positive control. The HA gel was synthesized as described in Example 7 above.

TABLE-US-00026 TABLE 1 Drug Total amount Drug administration of drugs Group concentration on right ear (μg/mouse) Model / Xylene / Vehicle / Xylene + gel containing / HA (without peptide) DEX 10 μg/μL Xylene + Dex cream 400 HA gel 0.5 mg/g Xylene + ‘HA gel’ 40

[0328] The mice were sacrificed by cervical dislocation after 40 minutes. The left and right ears were cut off. A skin pouch (Electron Microscopy Sciences, Hatfield, Pa., USA) with a diameter of 8 mm was used to take a piece of the ear from the same site of both ears. The weights were recorded, and the swelling rates were calculated as follows:

Swelling rate=(right ear weight−left ear weight)/left ear weight×100%

and the results showed in Table 2 below.

TABLE-US-00027 TABLE 2 Model Vehicle DEX HA gel Mean 97.4 97% 45% 50% SD 0.03 0.35 0.19 0.25

[0329] The above results show that HA gel could reduce the xylene induced swelling.