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EP-4735017-A1 - SUBSTRATES COMPRISING ELASTIN-LIKE POLYPEPTIDES AND CALCIUM IONS

EP4735017A1EP 4735017 A1EP4735017 A1EP 4735017A1EP-4735017-A1

Abstract

The disclosure is directed towards polypeptide substrates and methods of synthesis thereof. Such substrates can be embedded with calcium ions from a number of ionic sources. These calcium-embedded, polypeptide substrates can be used to grow a variety of crystal structures including flower-shaped, onion-shaped, and needle-like crystal structures. As such, the disclosure is additionally directed towards methods of crystal growth from polypeptide substrates. Compositions of the disclosure can be used in a wide variety of medical and other applications.

Inventors

  • HASAN, Abshar
  • ELSHARKAWY, SHERIF AHMED ABDELSALAM
  • CHAVARRIA, ALVARO MATA

Assignees

  • Mintech-V, LLC

Dates

Publication Date
20260506
Application Date
20240628

Claims (20)

  1. 1. A composition comprising: a disordered polypeptide substrate; and calcium ions embedded within the polypeptide substrate, wherein the calcium ions enhance the formation of ordered structures in the polypeptide substrate compared to a comparative polypeptide substrate consisting essentially of the same polypeptide substrate but without the calcium ions.
  2. 2. A substrate according to claim 1 wherein the calcium ions are Ca 2+ .
  3. 3. A substrate according to claim 1, wherein the calcium ions are provided by CaCL.
  4. 4. A substrate according to claim 1, wherein the calcium ions are present in an amount of at least 0.001% by weight of the substrate.
  5. 5. A substrate according to claim 4, wherein the calcium ions are present in an amount of at least 0.005 - 1.5% by weight of the substrate.
  6. 6. A substrate according to claim 1, wherein the polypeptide is a pentapeptide Elastin- like-polyptide selected from the group consisting of Gly-X-X-X-X, X-Gly-X-X-X, X- X-Gly-X-X, X-X-X-Gly-X and X-X-X-X-Gly, (GXXXX, XGXXX, XXGXX, XXXGX, XXXXG), wherein X is any amino acid apart from proline.
  7. 7. A substrate according to claim 1, wherein the polypeptide substrate has a thickness of from 0.5 mm - 1.5 mm.
  8. 8. A process for forming an elastin-like polypeptide membrane according to claim 1, the process comprising the steps of: a) dissolving elastin-like polypeptides with a source of calcium ions and a solvent to form an ELP solution; and b) applying the solution onto a surface to form a membrane.
  9. 9. A process according to claim 8, wherein the ELP is present in an amount of from 1- 20% by weight of the solution.
  10. 10. A process according to claim 8, wherein the source of calcium ions may be present in an amount of 0.005 - 1.5% by volume of the solution.
  11. 11. A process according to claim 8, wherein the step a) further comprises the step of mixing the ELP solution with a cross-linker.
  12. 12. A process according to claim 11, wherein the cross-linker is hexamethyl diisocyanate.
  13. 13. A crystal formed from and at least partly embedded in a substrate according to claim 1.
  14. 14. A crystal according to claim 13, wherein the crystal is located at least partly inside the bulk of the polypeptide substrate.
  15. 15. A crystal according to claim 13, wherein the crystal is located partly inside the bulk of the polypeptide substrate and partly on the surface of the polypeptide substrate.
  16. 16. A crystal according to claim 13, wherein the crystal has a hierarchical structure.
  17. 17. A crystal according to claim 13, wherein the crystal comprises nanocrystals.
  18. 18. A crystal according to claim 17, wherein the nanocrystals are arranged in concentric layers.
  19. 19. A crystal according to claim 17, wherein the nanocrystals have a needle shape.
  20. 20. A crystal according to claim 19, wherein the needle shaped nanocrystals are located on the polypeptide substrate surface and orientated perpendicular to the polypeptide substrate surface.

Description

SUBSTRATES COMPRISING ELASTIN-LIKE POLYPEPTIDES AND CALCIUM IONS PRIORITY AND INCORPORATION BY REFERENCE [0001] This application claims priority to U.S. Provisional Application Serial No. 63/511,035, filed June 29, 2023, the entire disclosure of which is hereby incorporated by reference herein for any and all purposes. This application hereby incorporates by reference the entire disclosure of U.S. Patent Application Serial No. 17/588,579, filed on January 31, 2022, including the sequence listings by reference herein for any and all purposes. TECHNICAL FIELD OF INVENTION [0002] The present invention relates to polypeptide substrates incorporating calcium ions and processes for their formation. The present invention also relates to crystals produced from these substrates and processes for their production. BACKGROUND [0003] Elastin-like polypeptides (ELPs) are a type of protein-like molecule that consist of repeating pentapeptide sequences of Val-Pro-Gly-Xaa-Gly (VPGXG), where X is any amino acid apart from proline. These molecules undergo a phase transition at a certain transition temperature (Tt), which results in the transition from a soluble to an insoluble form. In solutions with a temperature lower than Tt, free polymer chains remain in an unordered state showing full hydration (the soluble form). In solutions with temperatures exceeding Tt, polymer chains show a more ordered structure (known as the P-spiral), stabilized by hydrophobic interactions and intramolecular type P structures increasing the association of polymer chains. [0004] ELP membranes are known to form crystals that mimic natural enamel as outlined in WO2017168183. [0005] However, ELP membranes are limited in their ability to access more varied crystal morphologies and form crystal structures inside the membrane. [0006] Therefore, there exists a need to address limitations of membranes existing in the art. SUMMARY [0007] It is to be understood that this summary is not an extensive overview of the disclosure. This summary is exemplary and not restrictive and it is intended to neither identify key or critical elements of the disclosure nor delineate the scope thereof. The sole purpose of this summary is to explain and exemplify certain concepts of the disclosure as an introduction to the following complete and extensive detailed description. [0008] The present disclosure relates to a polypeptide substrate which can form crystal structures on both the interior and exterior of the membrane. Such substrates can be embedded with calcium ions. [0009] The present disclosure relates to a polypeptide substrate which can form more varied crystal structures including flower-shaped, onion-shaped, and needle-like crystals. [0010] The present disclosure relates to providing crystals having improved stiffness, toughness, hardness, wear resistance, compressive strength, and acid resistance. The present disclosure relates to a process of growing the mineralized structures epitaxially from the surrounding enamel or dentine tissues or other underlying crystal structures. Underlying crystal structures include bone tissue and surrounding dental enamel. This is facilitated by the incorporation of calcium ions into polypeptide substrates. [0011] The present disclosure relates to a process for forming a crystal structure from a polypeptide substrate incorporating calcium ions. [0012] The present disclosure relates to enhanced formation of amyloid-like ensembles from polypeptide molecules using Ca ions. [0013] The present disclosure relates to incorporating polypeptide substrates with or without crystal structures into a variety of devices and applications. [0014] The present disclosure relates to methods and compositions for overcoming or mitigating at least one problem of the prior art, whether expressly disclosed herein or not. BRIEF DESCRIPTION OF THE DRAWINGS [0015] The features and components of the following figures are illustrated to emphasize the general principles of the present disclosure. Corresponding features and components throughout the figures can be designated by matching reference characters for the sake of consistency and clarity. [0016] FIGS. 1A-1E display onion-shaped crystal structures according to aspects of the present disclosure. [0017] FIGS. 2A-2H display different stages of onion-shaped crystal growth according to aspects of the present disclosure. [0018] FIGS. 3A-3B display topography of perpendicular spiky crystal structures at varied calcium ion concentrations. [0019] FIGS. 4A-4G display flower-shaped crystal structures according to aspects of the present disclosure. [0020] FIGS. 5A-5D display crystal fusion inside polypeptide membranes according to the present disclosure. [0021] FIGS 6A-6D display mineralization (e.g., crystallization) occurring within thicker membrane cross-sections according to the present disclosure. [0022] FIGS. 7A-7B display crystal structure formation on elastin-like polypeptide microparticle substrates