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EP-4739277-A1 - CONSENSUS PEPTIDES IN HYA

EP4739277A1EP 4739277 A1EP4739277 A1EP 4739277A1EP-4739277-A1

Abstract

Pharmaceutical and/or cosmetic formulations are described and their uses in medicine comprising artificial peptides characterized by a proline sequence prevalent in key matricellular proteins that partake in wound healing as well as in bone and cartilage formation and connective tissue maintenance in all vertebrates. The current invention relates to a pharmaceutical formulation in the form of a gel comprising a. an artificial peptide comprising the amino acid sequence of Pro-X-X-Pro-Y-Y-Y-Pro-X-X-Pro-Y-Y-Pro-X-X-Pro-X-Pro-Y-Y-Y-Y-Y-Y-Pro-Y-Y-Y-Y-Y-Y-Pro-X-X-Pro-X-Pro-Y-Y-Y-Pro-Y-Y-Pro-Y-Pro-X-X-Pro-Y-Pro-Y-Y-Pro-X-X- Pro-Y-Y-Pro-X-X-Pro-Y-Y- Pro-X-X-Pro-Y-Pro-Pro-X-Pro-Pro-X-X-X-X-X-X-X-X-Pro-X-X-Pro-X-X-X-X (SEQ ID NO 1 ) and/or an artificial peptide comprising or consisting of the amino acid sequence of Pro-X-X-Pro-Y-Y-Pro-X-X-Pro-Y-Y- Pro-X-X-Pro-Y-Y- Pro-Y-Pro-Pro-X-Pro-Pro (SEQ ID NO 2), wherein i) Pro is proline; ii) X is an amino acid selected from the group consisting of Ala, lie, Leu, Met, Phe, Trp and Vai; iii) Y is an amino acid selected from the group consisting of Asn, Cys, Gin, Ser, Thr and Tyr, b. cross-linked Hyaluronic fibres (HA-XL), and c. linear hyaluronic acid fibres (HA).

Inventors

  • ØVREBØ, Øystein
  • ZAIDI, Rija Fatima
  • PERALE, GIUSEPPE
  • LYNGSTADAAS, Ståle Petter

Assignees

  • NuPep AS

Dates

Publication Date
20260513
Application Date
20240705

Claims (20)

  1. 1. A pharmaceutical and/or cosmetic formulation in the form of a gel comprising a. an artificial peptide comprising, or consisting of, the amino acid sequence: Pro-X-X-Pro-Y-Y-Y-Pro-X-X-Pro-Y-Y-Pro-X-X-Pro-X- Pro-Y-Y-Y-Y-Y-Y- Pro-Y-Y-Y-Y-Y-Y-Pro-X-X-Pro-X-Pro-Y-Y-Y-Pro-Y-Y-Pro-Y-Pro-X-X-Pro-Y- Pro-Y-Y-Pro-X-X- Pro-Y-Y-Pro-X-X-Pro-Y-Y-Pro-X-X-Pro-Y-Pro-Pro-X-Pro- Pro-X-X-X-X-X-X-X-X-Pro-X-X-Pro-X-X-X-X (SEQ ID NO 1), wherein i) Pro is proline; ii) X is an amino acid selected from the group consisting of Ala, lie, Leu, Met, Phe, Trp and Vai; iii) Y is an amino acid selected from the group consisting of Asn, Cys, Gin, Ser, Thr and Tyr, and b. 1-40mg/mL linear hyaluronic acid fibres (HA) and 1-40 mg/mL cross-linked hyaluronic acid fibres (HA-XL).
  2. 2. A pharmaceutical and/or cosmetic formulation in the form of a gel comprising a. an artificial peptide comprising, or consisting of, the amino acid sequence: Pro-X-X-Pro-Y-Y-Pro-X-X-Pro-Y-Y- Pro-X-X-Pro-Y-Y- Pro-Y-Pro-Pro-X- Pro-Pro (SEQ ID NO 2), wherein i) Pro is proline; ii) X is an amino acid selected from the group consisting of Ala, lie, Leu, Met, Phe, Trp and Vai; and iii) Y is an amino acid selected from the group consisting of Asn, Cys, Gin, Ser, Thr and Tyr, and b. 1-40mg/mL linear hyaluronic acid fibres (HA) and 1-40 mg/mL cross-linked hyaluronic acid fibres (HA-XL).
  3. 3. A pharmaceutical and/or cosmetic formulation according to claim 1 or 2, wherein the artificial peptide is at least 90% identical to an artificial peptide selected from the group consisting of the amino acid sequences of SEQ ID NO 1 , SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, and SEQ ID NO 5, such as wherein the artificial peptide is one or more artificial peptide(s) selected from the group consisting of artificial peptides comprising the amino acid sequence of SEQ ID NO 4 and SEQ ID NO 5.
  4. 4. A pharmaceutical and/or cosmetic formulation according to any of the preceding claims, comprising a. 0.1-250 pg/mL, such as 0.1 , 1.0, 5.0, 10, 50, 100, 200 or 250 pg/mL of said artificial peptide, b. 1-40 mg/mL, such as1.0, 2.5, 4, 10, 20, 25, 30 or 40 mg/mL cross-linked hyaluronic acid fibres (HA-XL), and c. 1-40 mg/mL, such as 1.0, 2.5, 4, 10, 20, 25, 30 or 40 mg/mL linear hyaluronic acid fibres (HA).
  5. 5. A pharmaceutical and/or cosmetic formulation according to any one of the preceding claims, wherein the cross-linked hyaluronic fibres (HA-XL) comprise or consist of 1 ,4-butanediol diglycidyl ether cross-linked hyaluronic acid fibres (HA- XL(BDDE)) and/or poly(ethylene glycol) diglycidyl ether (PEGDE) cross-linked hyaluronic acid fibres (HA-XL(PEGDE)).
  6. 6. A pharmaceutical and/or cosmetic formulation according to any of the preceding claims comprising a. 50 pg/mL of an artificial peptide with the amino acid sequence of SEQ ID NO 4 and/or SEQ ID NO 5, b. 20 mg/mL 1 ,4-butanediol diglycidyl ether cross-linked hyaluronic acid fibres (HA-XL(BDDE)) and/or poly(ethylene glycol) diglycidyl ether cross-linked hyaluronic acid fibres (HA-XL(PEGDE)), and c. 2.5 mg/mL linear hyaluronic acid fibres (HA).
  7. 7. A pharmaceutical and/or cosmetic formulation according to any of the preceding claims comprising a. 2 pg/mL of an artificial peptide with the amino acid sequence of SEQ ID NO 4 and/or SEQ ID NO 5, b. 20 mg/mL 1 ,4-butanediol diglycidyl ether cross-linked hyaluronic acid fibres (HA-XL(BDDE)) and/or poly(ethylene glycol) diglycidyl ether cross-linked hyaluronic acid fibres (HA-XL(PEGDE)), and c. 2.5 mg/mL linear hyaluronic acid fibres (HA). 8. A pharmaceutical and/or cosmetic formulation according to any of the preceding claims, wherein the hyaluronic acid fibres are between 0.7-4.0 MDa, such as between 1 .0-2.0, between 1.5-1 .
  8. 8, or between 3.0-3.3 MDa, such as 1.5MDa.
  9. 9. A pharmaceutical and/or cosmetic formulation according to claim 6, 7 or 8, further comprising d. one or more buffering agent(s) e. a source of fluoride, f. one or more salt(s), and g. water.
  10. 10. A pharmaceutical and/or cosmetic formulation according to claim 6, 7 or 8, further comprising d. Water, e. Sodium Fluoride (NaF), f. Citric Acid, g. Sodium Hydroxide, h. Sodium Chloride, i. Disodium Phosphate and j. Sodium Phosphate.
  11. 11 . A pharmaceutical and/or cosmetic formulation according to any of the preceding claims, wherein the artificial peptide is released at a controlled rate of 0.01-10 ug pr hour, such as about 0.01 , 0.1 , 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10 ug per hour.
  12. 12. A pharmaceutical and/or cosmetic formulation according to any of the preceding claims, wherein the composition further comprises Mesenchymal Stromal Cells (MSCs).
  13. 13. A pharmaceutical and/or cosmetic formulation according to claim 12, wherein the composition comprises between 100.000-10.000.000 MSCs pr. ml, preferably about 1.000.000 MSCs pr. ml.
  14. 14. A pharmaceutical formulation according to any of claims 1-13, wherein the composition promotes MSC homing.
  15. 15. A pharmaceutical formulation according to any of the preceding claims for use as a medicament.
  16. 16. A pharmaceutical formulation according to any of the preceding claims for use in soft tissue healing, for inducing neovascularization, for inducing reepithelization, for stimulating collagen production, for promoting oriented collagen formation, for graft taking, post-grafting, for chronic wound healing, for use in an antiinflammatory and/or antimicrobial treatment, and/or for use in treating periodontitis, periimplantitis, peri-mucositis, gingivitis, aphthous stomatitis and/or infections and/or inflammations in the soft tissue of the craniomaxillofacial complex.
  17. 17. A pharmaceutical formulation according to any of claims 1-13, for use as an antiinflammatory composition.
  18. 18. A pharmaceutical formulation for use according to claim 17, wherein the antiinflammatory effect is measured as a reduction in proinflammatory cytokines.
  19. 19. A pharmaceutical formulation for use according to claim 18, wherein the proinflammatory cytokines comprise one or more cytokines selected from the group consisting of IL-23, IL-1 alpha, IL-1 beta, TNF-alpha, MCP-1 , IL-12P70, IFN-Y, IFN-beta, IL-6, IL-10, IL-27, IL-17A and GM-CSF.
  20. 20. A method for producing a pharmaceutical and/or cosmetic formulation according to any of the preceding claims, said method comprises, a. providing an artificial peptide as defined in any one of claims 1-3, b. providing hyaluronic acid with a molecular weight of 0.7-4 MDa, such as between 1 .0-2.0, between 1.5-1 .8, or between 3.0-3.3 MDa, such as 1.5MDa, c. mixing 0.1-250 pg/mL of said artificial peptide and 1-40 mg/mL of said HA and d. optionally, adding a source of fluoride to said mixture, wherein, said pharmaceutical and/or cosmetic formulation has an osmolarity of 50- 400 mOsm/L, such as between 100 and 310 mOsm/L, or such as between 125 and 175 mOsm/L, such as about 150 mOsm/L, or such as between 275 and 325 mOsm/L, such as about 300 mOsm/L.

Description

CONSENSUS PEPTIDES IN HYA TECHNICAL FIELD The present invention relates to the field of soft tissue healing and/or improvement. Pharmaceutical and/or cosmetic formulations are described comprising artificial peptides characterized by a proline sequence prevalent in key matricellular proteins that partake in wound healing as well as in bone and cartilage formation and connective tissue maintenance in all vertebrates and their uses in medicine. The artificial peptides are herein formulated in hydrogels, comprising combinations of hyaluronic acids and cross- linked hyaluronic acids for minimally invasive administration and/or controlled release, for use in the treatment of soft tissue conditions, diseases and/or disorders, as well as for cosmetic uses, such as improved skin elasticity and/or hydration. BACKGROUND Soft tissue is a collective term for all the tissue in the body that is not hardened by the processes of ossification or calcification, such as bones and teeth. Soft tissue connects, surrounds or supports internal organs and bones, and includes muscle, tendons, ligaments, fat, fibrous tissue, lymph and blood vessels, fasciae, and synovial membranes. The term “soft tissue” is commonly used to describe muscles, tendons, ligaments and/or fascia, but several other tissue types and body systems contain soft tissue as well, including fat, skin, nerves, and blood vessels. Soft tissue disorders, diseases and damages, are medical conditions affecting soft tissue. They include trauma, wounds and soft tissue injuries to connective and/or epithelial tissue. In the craniomaxillofacial complex, soft tissue disorders include periodontitis, periimplantitis, peri-mucositis, gingivitis, aphthous stomatitis and other oral infections and/or inflammations. Skin and soft tissue infections remain among the most frequently encountered infections in surgery, and their severity ranges from mild cellulitis to severe, necrotizing infections with high incidences of morbidity and mortality. Most commonly, these disorders result from skin lesions in a susceptible host, but sometimes develop following hematogenous spread from a previously unknown focus. Biomaterial scientists design in particular organic bone substitutes based on the biochemical properties of the mimicked tissue to achieve near native functionality. Several non-collagenous proteins in bone are known as intrinsically disordered proteins (IDPs), as they lack detectible ordered domains and a fixed 3D structure under physiological conditions. EP2118136 discloses artificial IDPs peptides with improved properties for induction and/or stimulation of mineralization, in vivo and in vitro. Such peptides are provided which are easy to synthesize and methods of using the peptides for the induction and/or stimulation of mineral precipitation and/or biomineralization. The characterising sequence of amino acids of the artificial peptides disclosed in EP2118136 is a proline rich sequence prevalent in key matricellular proteins that partake in wound healing as well as in bone and cartilage formation and connective tissue maintenance in all vertebrates. This core sequence of prolines is highly conserved in vertebrates, is intrinsically disordered and does not induce any immunogenic response in humans. In contrast to the successful application of peptides for hard tissue healing, there is today still a largely unmet need for equally potent solutions for soft tissue healing and regeneration. In addition, because of the susceptibility of soft tissue to develop postsurgery and/or post-treatment complications and disorders, it would be preferable to find ways for minimally invasive administrations for the delivery of such advanced peptide- technology. Hydrogels are an extensively investigated class of biomaterials, and an increasing number of products have reached the clinic. Hydrogels represent a group of biomaterials consisting of water- swollen polymer or colloidal networks. Hydrogels are viscoelastic materials that have attracted attention in regenerative medicine due to their ability to structurally mimic the extracellular matrix (ECM), thereby creating a conducive environment for cell proliferation and tissue regeneration. The viscoelastic properties of hydrogels allow them to function as stem cell carriers or scaffolds for controlled drug release. There is a need for widely different applications for diverse tissues with different loading modes and levels and different clinical requirements of the material. Consequently, a one-fit-all hydrogel is an unlikely strategy. The current invention addresses both the need for advanced peptide-technology to promote healing, growth and differentiation of soft tissue, as well as the need for novel means of minimally invasive administration of such active substances to and/or into the soft tissue affected, in particular the soft tissue of the craniomaxillofacial complex. SUMMARY The current inventors have shown that the artificial IDPs pept