Sterilization of self-assembling peptides by irradiation

Patent metadata

Field	Value
Publication number	US-11534528-B2
Application number	US-202117217911-A
Country	US
Kind code	B2
Filing date	Mar 30, 2021
Priority date	Mar 31, 2020
Publication date	Dec 27, 2022
Grant date	Dec 27, 2022

Abstract

Gamma ray and e-beam irradiation provided efficient sterilization of certain self-assembling peptides (including RADA16 in solution) without substantial degradation of the major peptide, while, e.g., another self-assembly peptide, QLEL12 was significantly degraded following irradiation. Irradiation sterilization enhances the rheological property of, for example, RADA16 hydrogel once applied to tissue at a physiological pH. The rheological property increase can result in higher efficacy in a variety of biomedical applications.

First claim

The invention claimed is:

1. A method of sterilizing a self-assembling peptide (SAP) solution, the method comprising:
(a) placing one or more containers with the solution of self-assembling peptide into an irradiation machine, said self-assembling peptide selected from the group consisting of RADA16 (SEQ ID NO:1), KLD12 (SEQ ID NO:2), and IEIK13 (SEQ ID NO:3), and capable of forming a hydrogel when applied to a biological tissue at about neutral pH; and
(b) exposing the container to gamma ray, X-ray, and/or e-beam irradiation at a predetermined dose so that the self-assembling peptide solution is sterilized to a pre-determined Sterility Assurance Level (SAL) without substantial degradation of the peptide wherein the concentration of the degradation products of a full-length peptide in the solution post-irradiation ranges from 0.1% to 5% w/v while its desired biological or physical properties are maintained substantially at the same level or improved;
wherein the desired biological or physical properties are selected from the group consisting of: hemostatic, anti-adhesion, prevention of re-bleeding, anti-stenosis, tissue occlusion, mucosa elevation, wound healing, storage modulus, viscosity, and tissue void filling property.

2. The method of claim 1 , wherein the dose is 15-50 kGy.

3. The method of claim 1 , wherein the solution is irradiated by e-beam.

4. The method of claim 1 , wherein the peptide solution contains about 2.5% w/v of RADA16 (SEQ ID NO:1) and the dose is 15-24kGy.

5. The method of claim 1 , wherein the amount of total peptides that are degraded after the irradiation does not exceed 20% by weight of the amount of the self-assembling peptide prior to the irradiation.

6. The method of claim 5 , wherein the pre-irradiation bioburden of the self-assembling peptide solution is 9 CFU per product unit or less.

7. The method of claim 1 , wherein irradiation dose achieves sterility assurance level (SAL) of at least 10 −6 .

8. The method of claim 1 , wherein the pH of the solution pre- and post-irradiation ranges from about 1.8 to 3.5.

9. The method of claim 1 , wherein the one or more solutions containers is/are a plastic syringe(s).

10. The method of claim 1 , wherein the storage modulus of the hydrogel is increased at least by 10% post-irradiation.

11. The method of claim 1 , further comprising: c) shearing the solution to reduce or restore its storage modulus.

12. The method of claim 1 , wherein the self-assembling peptide solution exhibits a post-irradiation mass spectrometric (MS) profile having major M z peaks at 836/1670, 1100, and 1513 m/z.

Assignees

• 3 D Matrix Ltd

Inventors

• Rioult Marika G
• Gil Eun Seok
• Aleksi Elton
• Yamamoto Naoki

Classifications

• A61L27/227 Primary

  Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24 · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• A61K38/00

  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 title

• C07K7/08

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title

• A61L2/26

  Accessories · CPC title