Bonding together tissue with adhesive containing polyfunctional crosslinking agent and protein polymer

• US 5,817,303 A
• Filed: 05/05/1995
• Issued: 10/06/1998
• Est. Priority Date: 05/05/1995
• Status: Expired due to Term

• Current Assignee: Protein Polymer Technologies, Inc., Spine Wave Incorporated
• Original Assignee: Protein Polymer Technologies, Inc.
• Inventors: Stedronsky, Erwin R., Cappello, Joseph
• Primary Examiner(s): Naff, David M.
• Application Number:
  US08/435,641
• Time in Patent Office: 1,250 Days
• Field of Search: 435/174, 435/180, 424/486, 424/78.02, 514/2, 106/124, 106/124.1, 530/300, 530/350, 530/402, 530/810
• US Class Current: 424/78.02
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61L 24/106 Fibrin; Fibrinogen
• CPC Class Codes: A61L 24/108 Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 41/00 Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: Y10S 530/81 Carrier - bound or immobilized peptides or proteins and the preparation thereof, e.g. biological cell or cell fragment as carrier

Abstract

Proteinaceous polymers having repetitive units from naturally occurring structural proteins are employed as backbones for functionalities for crosslinking to provide strongly adherent tissue adhesives and sealants. Particularly, block copolymers having repeating units of elastin and fibroin are employed having lysine substitutions in spaced apart units, where the amino group can be crosslinked using difunctional crosslinking agents. The protein polymer contains at least 40 weight percent of repetitive units of 3 to 15 amino acids of at least one naturally occurring protein and in at least two units an amino acid is substituted by an amino acid containing a functional group capable of reacting with a crosslinking agent to form a strongly adherent adhesive composition for bonding together separated tissue or for sealing tissue defects. A preferred adhesive composition contains glutaraldehyde or polymethylene diisocyanate and a protein block copolymer of at least 30 kD containing at least 70 weight percent of repetitive units of Gly-Ala-Gly-Ala-Gly-Ser and Gly-Val-Gly-Val-Pro, where in at least two units an amino acid is substituted with lysine and the copolymer has a lysine equivalent weight in the range of 1 to 20 kD. The protein polymer is produced by recombinant DNA technology, and a kit can be formed containing the crosslinking agent and protein polymer.

Claims

1. A method of maintaining separated viable tissue in proximate relationship, said method comprising:

applying to said separated viable tissue to hold together said tissue when said tissue is in contiguous relationship, a precursor composition comprising a polyfunctional crosslinking agent and a protein polymer comprising at least 40 weight % of repetitive units of from 3 to 15 amino acids of at least one naturally occurring structural protein, where in at least two units an amino acid is substituted by an amino acid which comprises a functional group capable of reacting with said crosslinking agent, whereby said precursor composition sets up to a strongly adherent adhesive composition and said separated viable tissue is held in proximate relationship.

2. A method according to claim 1, wherein said functional group capable of reacting with said crosslinking agent is amino.

3. A method according to claim 1, wherein said functional group capable of reacting with said crosslinking agent is as a result of reacting a naturally occurring amino acid in said protein polymer with a polyfunctional compound without setting up of said composition.

4. A method according to claim 1, wherein said units are selected from the group consisting of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01), Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) and GXX, wherein X_-- is any amino acid, at least 10% and not more than 45% of X'"'"'s being proline.

5. A method according to claim 1, wherein said units are Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01) and Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) and wherein both of said units are present to form a block copolymer protein.

6. A method of maintaining separated viable tissue in proximate relationship, said method comprising:

applying to said separated viable tissue to hold together said tissue when said tissue is in contiguous relationship, a precursor composition comprising a polyfunctional crosslinking agent reactive with amino groups and a protein block copolymer comprising at least 70 weight % of repetitive units of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO;

01) and Gly-Val-Gly-Val-Pro (SEQ ID NO;

02), where in at least two units an amino acid is substituted with lysine, which lysine reacts with said crosslinking agent, whereby said precursor composition sets up to a strongly adherent adhesive composition and said separated viable tissue is held in proximate relationship.

7. A method according to claim 6, wherein the equivalent weight of said protein per lysine is in the range of 1 to 20 kD.

8. A method according to claim 6, wherein each block of said block copolymer has at least two units and the ratio of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01) to Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) is in the range of 1;

1-16.

9. A method according to claim 6, wherein said crosslinking agent has a plurality of amino reactive groups selected from the group consisting of aldehyde, isocyanate, thioisocyanate and activated carboxy.

10. A method according to claim 9, wherein said reactive group is aldehyde.

11. A method of maintaining separated viable tissue in proximate relationship, said method comprising:

applying to said separated viable tissue to hold together said tissue when said tissue is in contiguous relationship, a precursor composition comprising a polyfunctional crosslinking agent selected from the group consisting of glutaraldehyde and polymethylene diisocyanate and a protein block copolymer of at least 30 kD comprising at least 70 weight % of repetitive units of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO;

01) and Gly-Val-Gly-Val-Pro (SEQ ID NO;

02), where in at least two units an amino acid is substituted with lysine, said copolymer having a lysine equivalent weight in the range of 3 to 15 kD, whereby said precursor composition sets up to a strongly adherent adhesive composition and said separated viable tissue is held in proximate relationship.

12. A method of sealing a defect in viable tissue, said method comprising:

applying to said defect a precursor composition comprising a polyfunctional crosslinking agent and a protein polymer comprising at least 40 weight % of repetitive units of from 3 to 15 amino acids of at least one naturally occurring structural protein, where in at least two units an amino acid is substituted by an amino acid which comprises a functional group capable of reacting with said crosslinking agent, whereby said precursor composition sets up to a strongly adherent adhesive composition and said defect is sealed.

13. A method according to claim 12, wherein said units are selected from the group consisting of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01), Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) and GXX, wherein X_-- is any amino acid, at least 10% and not more than 45% of X'"'"'s being proline.

14. A method according to claim 12, wherein said units are Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01) and Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) and wherein both of said units are present to form a block copolymer protein.

15. A method according to claim 12, wherein said tissue is the wall of a vessel of the cardiovascular system.

16. A method of maintaining separated viable tissue in proximate relationship, said method comprising:

applying to said separated viable tissue to hold together said tissue when said tissue is in contiguous relationship, a precursor composition comprising a polyfunctional crosslinking agent and a protein polymer comprising at least 40 weight % of repetitive units of from 3 to 15 amino acids of at least one naturally occurring structural protein, where said protein polymer comprises at least two amino acids which comprise a functional group capable of reacting with said crosslinking agent, whereby said precursor composition sets up to a strongly adherent adhesive composition and said separated viable tissue is held in proximate relationship.

17. A kit comprising a polyfunctional crosslinking agent and a protein polymer comprising at least 40 weight % of repetitive units of from 3 to 15 amino acids of at least one naturally occurring structural protein, where in at least two units an amino acid is substituted by an amino acid comprising a functional group capable of reacting with said crosslinking agent.

18. A kit according to claim 17, wherein said functional group capable of reacting with said crosslinking agent is amino.

19. A kit according to claim 17, wherein said functional group capable of reacting with said crosslinking agent is as a result of reacting a naturally occurring amino acid in said protein polymer with a polyfunctional compound without setting up of said composition.

20. A kit according to claim 17, wherein said units are selected from the group consisting of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01), Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) and GXX, wherein X is any amino acid, at least 10% and not more than 45% of X'"'"'s being proline.

21. A kit according to claim 17, wherein said units are Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01) and Gly-Val-Gly-Val-Pro (SEQ ID NO;

02) and wherein both of said units are present to form a block copolymer protein.

22. A kit according to claim 17, wherein said crosslinking agent has a plurality of reactive groups selected from the group consisting of aldehydo, isocyanate, thioisocyante and activated carboxy.

23. A kit comprising a protein block copolymer of at least 30 kD comprising at least 70 weight % of repetitive units of Gly-Ala-Gly-Ala-Gly-Ser (SEQ ID NO:

01) and Gly-Val-Gly-Val-Pro (SEQ ID NO;

02), where in at least two units an amino acid is substituted with lysine, said copolymer having a lysine equivalent weight in the range of 1 to 20 kD and a crosslinking agent selected from the group consisting of glutaraldehyde and aliphatic diisocyanates.