Novel polypeptide and process for producing the same, and collagenase inhibitor

• US 20070207955A1
• Filed: 03/02/2006
• Published: 09/06/2007
• Est. Priority Date: 03/02/2006
• Status: Abandoned Application

• Current Assignee: Phg Corporation
• Original Assignee: Phg Corporation
• Inventors: Tanihara, Masao, Aoshima, Hisae
• Application Number:
  US11/365,936
• Publication Number:
  US 20070207955A1
• Time in Patent Office: Days
• US Class Current: 530/356
• CPC Class Codes: C07K 14/8146 Metalloprotease (E.C. 3.4.24) inhibitors, e.g. tissue inhibitor of metallo proteinase, TIMP

Abstract

The present invention provides a novel biodegradable and biosorbable polypeptide which is free from a risk of an infection by a pathogenic organism or a transmission of a causative factor, or a risk of an undesirable side effect, and a process for producing the same, as well as a collagenase inhibitor comprising the polypeptide and having a high collagenase inhibitory action. The polypeptide contains a peptide unit having an amino acid sequence represented by the following formula (1), and a peptide unit having an amino acid sequence represented by the following formula (2):
-Pro-X-Gly-  (1)
-Pro-Y-Gly-Z-Ala-Gly-  (2) wherein X represents Pro or Hyp; Y represents Gln, Asn, Leu, Ile, Val or Ala; and Z represents Ile or Leu. The molar ratio of the peptide unit (1) relative to the peptide unit (2) is about 99/1 to 1/99.

Claims

1. A novel polypeptide containing

a peptide unit having an amino acid sequence represented by the following formula (1), and

a peptide unit having an amino acid sequence represented by the following formula (2):

-Pro-X-Gly- 

 

(1)
-Pro-Y-Gly-Z-Ala-Gly- 

 

(2)

wherein X represents Pro or Hyp;

Y represents Gln, Asn, Leu, Ile, Val or Ala; and

Z represents Ile or Leu.

2. A polypeptide according to claim 1, wherein the proportion (molar ratio) of the peptide unit (1) relative to the peptide unit (2) is 99/1 to 1/99.

3. A polypeptide according to claim 1, wherein

(i) X is Hyp,

Y is Gln, Asn, Leu, Ile, Val or Ala, and

Z is Ile or Leu;

or

(ii) X is Pro,

Y is Gln, Asn, Leu, Ile, Val or Ala, and

Z is Ile or Leu.

4. A polypeptide according to claim 1, which is degradable with a collagenase.

5. A polypeptide according to claim 1, which shows positive Cotton effect at a wavelength in range of 220 to 230 nm and negative Cotton effect at a wavelength in range of 195 to 205 nm in a circular dichroism spectrum, and wherein at least part of the polypeptide is capable of forming a triple helical structure.

6. A polypeptide according to claim 1, which shows a peak of the molecular weight in the range from 5×

10² to 500×

10⁴ in the molecular weight distribution.

7. A polypeptide according to claim 1, which is capable of forming a collagenous tissue.

8. A process for producing a polypeptide recited in claim 1, which comprises condensing an amino acid component or peptide component which at least contains an amino acid or peptide fragment corresponding to the formula (1) recited in claim 1 and an amino acid or peptide fragment corresponding to the formula (2) recited in claim 1.

9. A process for producing a polypeptide recited in claim 1, which comprises

(a) condensing a peptide component which at least contains a peptide having the both amino acid sequences represented by the formulae (1) and (2) recited in claim 1, respectively;

or

(b) condensing a peptide component which at least contains a peptide having an amino acid sequence represented by the formula (1) and a peptide having an amino acid sequence represented by the formula (2).

10. A process according to claim 8 or 9, wherein the condensation step is carried out in the presence of (i) a dehydrating and condensing agent, or (ii) a dehydrating and condensing agent and a condensing auxiliary.

11. A process according to claim 10, wherein the dehydrating and condensing agent comprises at least one member selected from the group consisting of a carbodiimide-series condensing agent, a fluorophosphate-series condensing agent, and a diphenylphosphorylazide.

12. A process according to claim 10, wherein the condensation step is carried out in the presence of a non-aqueous solvent, and the proportion of the dehydrating and condensing agent is 0.7 to 5 mol relative to 1 mol of the total amount of the amino acid component or peptide component.

13. A process according to claim 10, wherein the condensation step is carried out in the presence of an aqueous solvent, and the proportion of the dehydrating and condensing agent is 2 to 500 mol relative to 1 mol of the total amount of the amino acid component or peptide component.

14. A process according to claim 10, wherein the condensing auxiliary comprises at least one member selected from the group consisting of an N-hydroxypolycarboxylic acid imide, an N-hydroxytriazole, a triazine, and ethyl ester of 2-hydroxyimino-2-cyanoacetic acid.

15. A process according to claim 10, wherein the proportion of the condensing auxiliary is 0.5 to 5 mol relative to 1 mol of the total amount of the amino acid component or the peptide component.

16. A collagenase inhibitor comprising a polypeptide recited in claim 1.

17. A cosmetic preparation comprising a polypeptide recited in claim 1, and inhibiting collagenase activity.

18. A food composition comprising a polypeptide recited in claim 1, and inhibiting collagenase activity.

19. A method for inhibiting a collagenase activity which comprises acting a polypeptide recited in claim 1 on a collagenase.