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US-20260124244-A1 - MEGAKARYOCYTE DERIVATIVES FOR TREATMENT OF DRY EYE DISEASES

US20260124244A1US 20260124244 A1US20260124244 A1US 20260124244A1US-20260124244-A1

Abstract

Described herein are methods for treating, repairing or ameliorating diseases, disorders, or injuries related to dry eye disease, wound healing, and osteoarthritis with megakaryocyte derivatives. Also, described herein are methods for generating megakaryocyte derivatives.

Inventors

  • Bradford DYKSTRA
  • Ryan Carpenter
  • Sunita PATEL-HETT
  • Sophia PETE

Assignees

  • STELLULAR BIO, INC.

Dates

Publication Date
20260507
Application Date
20250702

Claims (20)

  1. 1 .- 44 . (canceled)
  2. 45 . A method of treating a condition comprising: differentiating an ex-vivo culturing of a population of induced pluripotent stem cells (iPSCs) into mature megakaryocyte-like cells (MLCs); lysing the mature MLCs to produce lysates; processing the lysates to form a therapeutic composition, wherein the therapeutic composition comprises one or more of: epidermal growth factor (EGF)>10% higher or interleukin (IL)-10>10% higher as compared to plasma rich in growth factors (PRGF), or EGF>10% higher, IL-10>10% higher as compared to crude lysates; and administering to a subject suffering from a dry eye disease an effective amount of the therapeutic composition, thereby treating a dry eye disease.
  3. 46 . The method of claim 45 , wherein the lysates further comprise one or more of: EGF>50% higher or IL-10>50% higher as compared to PRGF.
  4. 47 . The method of claim 45 , wherein the lysates further comprise one or more of: matrix metalloproteinase (MMP)-1>10% higher, MMP-9>10% higher, MMP-12, tissue inhibitors of metalloproteinases (TIMP)-1>10% higher, TIMP-3>10% higher, transforming growth factors (TGF)-B1>10% higher, TGF-B2>10% higher, Endoglin>10% higher, Endothelin-3>10% higher, Follistatin>10% higher, heparin-binding (HB)-EGF>10% higher, hepatocyte growth factor (HGF)>10% higher, placental growth factor (PLGF)>10% higher, vascular endothelial growth factor (VEGF)-A>10% higher, soluble CD40L (sCD40L)>10% higher, Eotaxin>10% higher, fibroblast growth factors (FGF)-2>10% higher, FLT-3L>10% higher, Fractalkine>10% higher, granulocyte colony stimulating factor (G-CSF)>10% higher, growth-regulated oncogene a (GROa)>10% higher, interferon (IFN)-a2>10% higher, IFNγ>10% higher, IL-1a>10% higher, IL-1B>10% higher, IL-1RA>10% higher, IL-2>10% higher, IL-3>10% higher, IL-4>10% higher, IL-5>10% higher, IL-6>10% higher, IL-8>10% higher, IL-9>10% higher, IL-10>10% higher, IL-12p40>10% higher, IL-12p70>10% higher, IL-13>10% higher, IL-15>10% higher, IL-17A>10% higher, IL-17E>10% higher, IL-25>10% higher, IL-17F>10% higher, IL-22>10% higher, IL-27>10% higher, interferon gamma-induced protein (IP)-10>10% higher, monocyte chemoattractant protein (MCP)-1>10% higher, MCP-3>10% higher, macrophage-derived chemokine (MDC)>10% higher, macrophage inflammatory protein (MIP)-1a>10% higher, MIP-1b>10% higher, platelet-derived growth factor (PDGF)-AA>10% higher, transforming growth factor (TGF) a>10% higher, tumor necrosis factor (TNF) a>10% higher, TNFB>10% higher, a proliferation-inducing ligand (APRIL)>10% higher, B-cell activating factor (BAFF)>10% higher, regulated upon activation, normal T cell expressed and secreted (RANTES)>10% lower, IL-7>10% lower, VEGF-D>10% lower, VEGF-C>10% lower, Leptin>10% lower, angiopoietin (Ang) 2>10% lower, transforming growth factor (TGF) B-3>10% lower, MMP-10>10% lower, MMP-7>10% lower, MMP-3>10% lower, or MMP-2>10% lower as compared to PRGF as compared to PRGF.
  5. 48 . The method of claim 45 , wherein the lysates further comprise one or more of: MMP-7>10% higher, TIMP-1>10% higher, TIMP-3>10% higher, IL-2>10% higher, IL-15>10% higher, IL-17F>10% higher, granulocyte colony stimulating factor (G-CSF)>10% higher, MMP-13>10% lower, TGFB-1>10% lower, IL-5>10% lower, APRIL>10% lower, or BAFF>10% lower as compared to crude lysates.
  6. 49 . The method of claim 45 , wherein the method improves one or more signs and/or symptoms of dry eye disease.
  7. 50 . The method of claim 45 , wherein the dry eye disease is caused by Sjogren's syndrome or non-Sjogren's syndrome.
  8. 51 . The method of claim 45 , wherein the therapeutic composition is diluted to a physiological concentration in a carrier, wherein the carrier comprises a diluent or an excipient.
  9. 52 . The method of claim 45 , wherein the therapeutic composition further comprises a wound healing agent, a tissue regeneration agent, an antiapoptotic agent, an anti-inflammatory agent, a neurotropic agent, an anti-hormonal agent or an immunomodulatory agent or a combination thereof.
  10. 53 . The method of claim 52 , wherein the tissue regeneration agent is one or more of (a) a growth factor selected from one or more of transforming growth factors (TGF), fibroblast growth factors (FGF), platelet-derived growth factors (PDGF), epidermal growth factors (EGF), vascular endothelial growth factors (VEGF), insulin-like growth factors (IGF), platelet-derived endothelial growth factors (PDEGF), platelet-derived angiogenesis factors (PDAF), platelet factors 4 (PF-4), hepatocyte growth factors (HGF) or combinations thereof; and (b) a cytokine selected from one or more of IL-1B, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, IL-17A, IL-23, TNF alpha or combinations thereof.
  11. 54 . The method of claim 45 , wherein the population of cells comprises cells derived from immortalization, reprogramming or differentiation of a somatic cell, a progenitor cell or a stem cell.
  12. 55 . The method of claim 45 , wherein the therapeutic composition further comprises lysates, serum, plasma, plasma rich in growth factors (PRGF), platelet rich plasma (PRP), or any other blood derivative derived from the subject.
  13. 56 . The method of claim 45 , wherein the therapeutic composition in formulated in a buffer, diluent, or excipient or a combination thereof.
  14. 57 . The method of claim 45 , wherein the therapeutic composition comprises: a) between 0.01 and 100 wt % of the lysates, b) between 0 and 90 wt. % of a bulking agent, and/or c) between 0 and 90 wt. % of at least one excipient or carrier and optionally d) platelet-rich plasma (PRP), PRGF, plasma, serum, or other blood derivative derived from the subject.
  15. 58 . The method of claim 45 , wherein the therapeutic composition is lyophilized.
  16. 59 . The method of claim 45 , wherein the administering comprises locally administering the therapeutic composition at one or more of a site of or near an injury or a disease, wherein the therapeutic composition is administered by any one of route of administrations selected from topical, transdermal, or systemic route of administration, or wherein the therapeutic composition is administered by any one of route of administrations selected from subconjunctival, sub-Tenon's, intravitreal, intracameral, intravenous, intraarterial, intramuscular, subcutaneous, inhalation, rectal, buccal, vaginal, intraperitoneal, intraarticular, intraglandular into a lacrimal gland, topical, otic, or oral route of administration.
  17. 60 . A method of treating a condition comprising: administering to a subject suffering from a dry eye disease an effective amount of a therapeutic composition, the therapeutic composition comprising lysates of megakaryocyte-like cells (MLCs), wherein, after processing, the lysates comprise one or more of: epidermal growth factor (EGF)>10% higher or interleukin (IL)-10>10% higher as compared to plasma rich in growth factors (PRGF), or EGF>10% higher, IL-10>10% higher as compared to crude lysates.
  18. 61 . A method for producing a therapeutic composition comprising an effective amount of processed lysates of MLCs, the method comprising: an ex-vivo culturing of a population of iPSCs for a duration to differentiate the iPSCs to mature MLCs; concentrating the MLCs; lysing the MLCs; and processing the lysed MLCs to form the therapeutic composition; wherein, after processing, the therapeutic composition comprises one or more of: epidermal growth factor (EGF)>10% higher or interleukin (IL)-10>10% higher as compared to plasma rich in growth factors (PRGF), or EGF>10% higher, IL-10>10% higher as compared to crude lysates.
  19. 62 . The method of claim 61 , wherein the processing comprises filtering the lysed MLCs.
  20. 63 . The method of claim 61 , wherein the processing comprises centrifuging the lysed MLCs.

Description

CROSS-REFERENCED APPLICATION This application is a bypass continuation patent application of PCT International Patent Application No. PCT/US2024/010568, filed Jan. 5, 2024, which claims the benefit of and priority to U.S. Provisional Application No. 63/437,578, filed on Jan. 6, 2023, the entire contents of each of which are incorporated herein by reference. BACKGROUND Blood derivatives, such as platelet rich plasma (PRP), plasma rich in growth factors (PRGF), and autologous serum (AS) have been implicated to play a role in medical treatment of wound healing or speeding repairs of damaged tissues such as cartilage, tendons, ligaments and bone, or in treating dry eye and other diseases. However, there is a paucity of critical data regarding the beneficial effects of these blood derivatives in clinical procedures because of limited sample size of reported data from platelet donors. Moreover, every platelet donor is different, therefore, there is no consistency in the quality of PRP or other derivatives produced for medical procedures. Autologous blood derivatives, currently in use, are plagued with several other disadvantages, such as batch to batch variability because every patient provides his or her own blood as a source of PRP, PRGF, or AS. Other disadvantages include the short life-span of donor platelets, impurities or contaminations, or lack of availability or shortages in supply. As a result of a lack of standardization of these products, clinical effectiveness of blood derivatives is still under debate. Hence, the field is in dire need of a safe, standardized, defined, off-the-shelf product that could be developed and approved for use in humans by regulatory bodies, instead of having to make separate, unreliable preparations for each patient, which has raised skepticism regarding its potential efficacy and use. Dry eye disease (DED) is an example of a disease for which there is a significant unmet need, and a treatment is urgently desired. In the current DED treatment, patients are prescribed one of a number of treatments, for example Restasis®, which the active ingredient is Cyclosporin A (CsA). Unfortunately, current medications are not effective for many dry eye patients, and their disease often progresses. Dry eye diseases fall into two categories, which are (i) tear-deficient and (ii) evaporative. The evaporative causes of dry eye disease are due to oil deficit, lid changes, use of contact lenses, or ocular surface diseases (OSD), as allergic conjunctivitis, and some of the iatrogenic dry eye that occurs after the use of systemic or topical medications or after surgeries or nonsurgical procedures. If symptoms continue to progress, patients are deemed to require a surgery and are referred to an ophthalmologist. Hence, novel, less aggressive and better therapeutic options for the treatment of dry eye disease are needed and are described in this application. SUMMARY Provided herein are methods of treating, repairing or ameliorating dry eye disease in a subject in need of such treatment. In some embodiments, the methods comprise administering to the subject an effective amount of a composition comprising a megakaryocyte derivative, wherein an administration route is ophthalmic or intraglandular, wherein the megakaryocyte derivative includes megakaryocyte-like cell (MLC) lysates, platelet-like cells (PLC), or combinations thereof. Provided herein are methods of treating, repairing or ameliorating a condition in a subject in need of such treatment comprising administering to the subject more than one dose of an effective amount of a composition comprising a megakaryocyte derivative, wherein the condition is selected from dry eye disease, ocular surface disease, osteoarthritis, and wound healing, and wherein the megakaryocyte derivative includes MLC lysates, PLCs, exosomes, megakaryocytes, or combinations thereof, and wherein the megakaryocyte derivative comprises one or more biomarkers selected from fibroblast growth factor-2 (FGF-2); hepatocyte growth factor (HGF); insulin-like growth factor 1 (IGF-1), regulated upon activation, normal T cell expressed and secreted (RANTES); nerve growth factor (NGF); vascular endothelial growth factor (VEGF-A); vascular endothelial growth factor (VEGF-C); epidermal growth factor (EGF); transforming growth factor-β1 (TGF-β1); transforming growth factor-β2 (TGF-β2); platelet-derived growth factor-AA (PDGF-AA); platelet-derived growth factor-BB (PDGF-BB); platelet-derived growth factor-AA/BB (PDGF-AA/BB); interleukin 2 (IL-2); interleukin 4 (IL-4); interleukin 12p40 (IL-12p40); interleukin 12p70 (IL-12p70); tissue inhibitors of metalloproteinases-1 (TIMP-1); tissue inhibitors of metalloproteinases-2 (TIMP-2); and tissue inhibitors of metalloproteinases-3 (TIMP-3). Provided herein are method for producing megakaryocyte derivatives, the methods comprise: an ex-vivo culturing of a population of progenitor cells for a duration to differentiate the progenitor cells to mature