Methods to treat fibrotic diseases, disorders or conditions using adipogenic promoting factors

Abstract

Provided are methods for treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof by administering a pharmaceutically acceptable composition comprising one or more pro-adipogenic factors that transform or reprogram fibroblasts or myofibroblasts into adipocyte-like cells. Also provided are methods to transform or reprogram fibroblasts or myofibroblasts into adipocyte-like cells by contacting fibroblasts or myofibroblasts with a composition comprising ONO 1301 or (Z)-ONO 1301.

Description

Atorney docket No. 00058-095W01METHODS TO TREAT FIBROTIC DISEASES, DISORDERS OR CONDITIONS USING ADIPOGENIC PROMOTING FACTORS CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority under 35 U.S. C. §119 from Provisional Application Serial No. 63 / 780,165, filed March 28, 2025, the disclosure of which is incorporated herein by reference.TECHNICAL FIELD

[0002] The disclosure provides methods for treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof by administering a pharmaceutically acceptable composition comprising one or more pro-adipogenic factors that reprogram fibroblasts or myofibroblasts into adipocyte-like cells.BACKGROUND

[0003] Dupuytren disease (DC) is a common fibrotic condition of the palm and digits. It may lead to flexion deformities of the digits and impaired hand function known as Dupuytren contracture (DC). Myofibroblasts are the primary cells responsible for the development of the disease. Surgical treatments encompassing a variety of approaches, percutaneous release, and collagenase Clostridium histolyticum (CCH) are all standard treatments for Dupuytren disease. However, although these modalities have a reasonable safety profile, the tendency for recurrence with all modalities is high and usually occurs within 2 years of treatment.SUMMARY

[0004] Dupuytren’s disease is a fibroproliferative disorder that affects the palm, causing a stiff, persistent flexion of the fingers due to contracting fibrous cords. The pathogenesis of Dupuytren’s involves pathologic myofibroblasts in the palmar fascia that secrete abnormal collagens and induce contraction. The standard surgical treatment is fasciectomy. However, this surgery is associated with a significant recovery time, necessitates skilled therapy, and has an undesirable complication profile. The FDA approved collagenase Clostridium histolyticum (CCH) as a non-operative treatment for DC. However, even though this is safe and non-operative, the tendency of recurrence is within 2 years. Thus, novel therapeutic options are needed.

[0005] The reprogramming of myofibroblasts into adipocytes represents a means to prevent or treat fibrosis. Identified herein are pro-adipogenic factors which act to reprogram fibroblasts and myofibroblasts into adipocyte-like cells. The disclosure provides methods forAtorney docket No. 00058-095W01treating subjects with Dupuytren disease comprising administering therapeutically effective amounts of these pro-adipogenic factors disclosed herein. The methods of treatment disclosed herein would reduce the need for moderate or radical fasci ectomy and limit the surgery to a percutaneous contracture release, and growth factor or small molecule injection, aimed at reducing fibrosis and replenishing subdermal fat loss and reducing recurrence. Accordingly, the methods of disclosure provide a small molecule solution to treat Dupuytren disease, instead of the current surgical and cellular therapeutic treatments. The pro-adipogenic factors disclosed herein can be administered as an injectable therapeutic, optionally with fat graft components and / or antifibrotic treatments. Additionally, the pro-adipogenic factors disclosed herein can be used as a topical formulation or microneedle patch formulation as the molecules are low molecular weight allowing for percutaneous absorption.

[0006] In a particular embodiment, the disclosure provides a method to treat Dupuytren disease in a subject in need thereof, comprising: administering therapeutically effective amounts of pro-adipogenic factors to the subject in need thereof, either alone or in combination. In a further embodiment, the combination of tw o or more pro-adipogenic factors includes IGF-1 or an IGF-1 related agent, and HGF or a HGF related agent. In yet a further embodiment, the combination of two or more pro-adipogenic factors includes IGF-1 and HGF. In another embodiment, the combination of two or more pro-adipogenic factors includes at least one IGF-1 related agent, and at least one HGF related agent. In yet another embodiment, the at least one IGF-1 related agent comprises des-(l-3) IGF-I (des-IGF-I). glycine-proline-glutamate (GPE), and / or cyclic gly cine-proline (cGP). In a further embodiment, the at least one IGF-1 related agent comprises cGP. In a certain embodiment, the at least one HGF related agent comprises dihexa and / or ONO-1301. In another embodiment, the two or more pro-adipogenic factors comprises cGP. dihexa, and ONO- 1301. In yet another embodiment, the combination of two or more pro-adipogenic factors are formulated for sterile injection. In a further embodiment, the combination of two or more pro-adipogenic factors are formulated for topical administration. In yet a further embodiment, the combination of two or mor pro-adipogenic factors act in a synergistic manner to reprogram myofibroblasts into adipocytes. In another embodiment, the method is used in combination with one or more Dupuytren disease treatments.

[0007] In a particular embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprisingAtorney docket No. 00058-095W01therapeutically effective amounts of one or more pro-adipogenic factors that transform or reprogram fibroblasts or myofibroblasts into adipocyte-like cells. In a further embodiment, the fibrotic disease, disorder or condition is associated with fascia fibrosis. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition is selected from plantar fibroma, Dupuytren's disease, adhesive capsulitis, Peyronie's disease, eosinophilic fasciitis, scleroderma, and chronic myofascial pain syndrome. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition is Dupuytren's disease. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition is associated with cutaneous fibrosis. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the cutaneous fibrosis results from abnormal matrix or fibroblast accumulation, neoplasia, injury-induced fibrosis, or immune dysregulation. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition results from abnormal matrix or fibroblast accumulation in skin and is selected from stiff skin syndrome, Winchester syndrome, hyaline fibromatosis syndrome, Buschke-Ollendorf syndrome, Hutchinson-Gilford progeria syndrome, restrictive dermopathy, and atypical Wemer syndrome. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition results from neoplasia in skin and is selected from dermatofibrosarcoma protuberans, desmoid tumor, fibrous hamartoma of infancy, infantile myofibromatosis, Proteus syndrome, sclerotic fibroma, dermatofibroma, and other fibrohistiocytic / fibroblastic / myofibroblastic tumors and sarcomas. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition results from injury’ -induced fibrosis in skin and is selected from bum, physiologic scar, hypertrophic scar, keloidal scar, radiation-induced fibrosis, nephrogenic systemic fibrosis, knuckle pads, lipodermatosclerosis, and dermatofibroma. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease, disorder or condition results from immune dysregulation in skin and is selected from lichen sclerosus, morphea, scleroderma, scleredema, scleromyxedema, sclerodermatous GVHD, eosinophilic-myalgia syndrome, toxic oil syndrome, acrodermatitis chronica atrophicans, acne keloidalis nuchae, and dermatofibroma. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic disease or condition is associated with fibrosis in an organ selected from lung, liver, heart and kidney. InAtorney docket No. 00058-095W01another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibrotic diseases or condition is selected from idiopathic pulmonary fibrosis, fibrosis of the liver, liver cirrhosis, cardiac fibrosis, and renal fibrosis. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the subject is a human patient. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the subject is anon-human subject. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition further comprises a pharmaceutically acceptable diluent, excipient and / or carrier. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition is formulated for oral administration. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition is formulated for subcutaneous administration. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition is formulated for topical administration. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition is formulated for administration by a microneedle patch. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition further comprises one or more anti-fibrotic medications. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more anti-fibrotic treatments are selected from nintedanib, pirfenidone. verapamil, collagenase, corticosteroids, mycophenolate mofetil, methotrexate, rituximab, ruxolitinib, belumosudil, celastrol, 5-fluorouracil, autologous fat grafting, tocilizumab, and triamcinolone. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the method further comprises administering to the subject one or more anti-fibrotic treatments. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more anti-fibrotic treatments are administered concurrently or sequentially with the pharmaceutically acceptable composition. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more anti-fibrotic treatments are selected from nintedanib, pirfenidone, verapamil, collagenase, corticosteroids, mycophenolate mofetil, methotrexate, rituximab, ruxolitinib, belumosudil, celastrol, 5-fluorouracil, autologous fat grafting, tocilizumab, and triamcinolone. In another embodiment, or in a further embodiment of any of the foregoing embodiments, at least one of the pro-adipogenic factors is ONO 1301 having the structure ofAtorney docket No. 00058-095W01. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises ONO 1301 at a concentration from 10 pM to 100 pM. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises ONO 1301 at a concentration from 40 pM to 70 pM. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises ONO 1301 at a dose of 4 mg / kg to 50 mg / kg. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises ONO 1301 at a dose of 10 mg / kg to 30 mg / kg. In another embodiment, or in a further embodiment of any of the foregoing embodiments, ONO 1301 is polymerized with poly-lactic co-glycolic acid (PLGA) microspheres to provide sustained or extended release of ONO 1301 when administered. In another embodiment, or in a further embodiment of any of the foregoing embodiments, at least one of the pro-adipogenic factors is (Z)-ONO 1301 having the structureN . In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a concentration from 10 pM to 100 pM. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a concentration from 40 pM to 70 pM. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a dose of 5 mg / kg to 200 mg / kg. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a dose of 10 mg / kg to 50 mg / kg. In another embodiment, or in a further embodiment of any of the foregoing embodiments, (Z)-ONO 1301 is polymerized with polyAtorney docket No. 00058-095W01lactic co-glycolic acid (PLGA) microspheres to provide sustained or extended release of (Z)-ONO 1301 when administered. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more pro-adipogenic factors comprises at least Insulin-like Growth Factor 1 (IGF-1) or an IGF-1 -related agent, and at least hepatocyte growth factor (HGF) or an HGF -related agent. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more pro-adipogenic factors comprises IGF-1 and HGF. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more pro-adipogenic factors comprises at least an IGF-1 -related agent, and at least an HGF-related agent. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the at least one IGF-1 -related agent comprises des-(l-3) IGF-I (des-IGF-I), glycine-proline-glutamate (GPE). and / or cyclic gly cine-proline (cGP). In another embodiment, or in a further embodiment of any of the foregoing embodiments, the at least one IGF-l-related agent comprises cGP. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the at least one HGF-related agent comprises ONO-1301 and / or dihexa having the structure ofNHj / G. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the one or more pro-adipogenic factors comprises cGP, dihexa, and ONO-1301. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the method is used in combination with one or more Dupuytren disease treatments. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the Dupuytren disease treatments is selected from collagenase injections or needle aponeurotomy, surgical fasciotomy, steroid injections, radiation therapy, and fas ci ectomy.

[0008] In a particular embodiment, the disclosure also provides a method to transform or reprogram fibroblasts or myofibroblasts into adipocytes, comprising: contacting fibroblasts or myofibroblasts with a composition comprising ONO- 1301 having the structure of:Atorney docket No. 00058-095W01of any of the foregoing embodiments, ONO-1301 or (Z)-ONO 1301 is used at a concentration from 10 pM to 100 pM. In another embodiment, or in a further embodiment of any of the foregoing embodiments, ONO-1301 or (Z)-ONO 1301 is used at a concentration from 40 pM to 70 pM. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibroblasts or myofibroblasts are contacted in vitro. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibroblasts or myofibroblasts are contacted in vivo. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the fibroblasts or myofibroblasts are myofibroblasts of a fibrotic disease, disorder or condition. In another embodiment, or in a further embodiment of any of the foregoing embodiments, the myofibroblasts are Dupuytren myofibroblasts.

[0009] In a certain embodiment, the disclosure provides a composition, a method or a kit as substantially described herein.DESCRIPTION OF DRAWINGS

[0010] FIG. 1 presents a Dupuytren animal model and in vitro studies with Dupuytren myofibroblasts (MFs). Phase I, in vivo studies, presents the treatment and timing schedule for mu / mu male rats that have been implanted with Dupuytren MFs or normal human dermal fibroblasts into the paws. Phase II, in vitro studies, Dupuytren MFs were treated and assessed for adipocyte formation.

[0011] FIG. 2A-E presents a model of Dupuytren disease in vivo. (A) Normal human dermal fibroblasts and Dupuytren MFs were implanted into the paws of mu / mu rats. After approximately 2 months, the paw tissue was harvested. Masson’s tri chrome and IHC staining was conducted to detect Collagen ty pe I, Collagen type III and a-SMA (n = 3 / group).Representative images are shown and labeled. Scale bar = 100 pm. ImmunohistochemistryAtorney docket No. 00058-095W01staining was quantified using ImageJ for (B) Collagen type I and (C) Collagen type III. (D) The ratio of Collagen type III to Collagen type I is presented. (E) Immunohistochemistry (IHC) staining was quantified using ImageJ for a-SMA. The data are presented as the mean ± SD. *P < .05. AU; arbitrary units.

[0012] FIG. 3A-E shows adipose-derived stem cells (ASC) + platelet rich plasma (PRP) treatment reduces fibrosis markers in vivo. Dupuytren MFs were implanted into the paws of mu / mu rats. After approximately 2 months, the paw tissue was treated with saline, ASC + PRP, or CCH once a week for 3 weeks. The paw tissue was harvested 1 week after each treatment. (A) Masson’s trichrome was conducted. A represented image of saline-treated Dupuytren MF-implanted paws served as a comparison. Scale bar = 100 pm.Immunohistochemistry was performed to detect (B) Collagen type I and (C) Collagen type III. (D) The ratio of Collagen type III to Collagen type I is presented. (E) IHC staining was performed to detect a-SMA. ImageJ was used to quantify the staining (n = 3 / group). The data are presented as the mean ± SD. *P < .05; **P < .01. AU; arbitrary units.

[0013] FIG. 4 presents ASC + PRP treatment induces adipocyte reemergence in vivo. Dupuytren MFs were implanted into the paws of mu / mu rats. After approximately 2 months, the paw tissue was treated with saline, ASC + PRP, or collagenase Clostridium histolyticum (CCH) once a week for 3 weeks. The paw tissue was harvested 1 week after each treatment. Immunohistochemistry was performed to detect perilipin. Perilipin served as a marker of adipocytes. Adipocytes were counted using ImageJ and the Adipocyte Tools plugin. The data are presented as the mean ± SD. *P < .05; **P < .01.

[0014] FIG. 5 provides Dupuytren MFs treated with ASC + Insulin-like grow th factor-1 (IGF-1) transform into lipid-filled cells. Dupuytren MFs were left untreated, cocultured with ASCs and PRP (1%) as a positive control, treated with IGF-1 (100 ng / mL), or treated with ASCs + IGF-1 (100 ng / mb). The cells were fixed, stained with Oil Red O, and imaged at magnification x 10. Representative images are shown. The scale bar represents 50 pm.

[0015] FIG. 6 provides the results of an IGF-1 escalating dose experiment in lipid-filled cells, to discover an optimal dose of IGF-1, which was found to be 100 ng / mL.

[0016] FIG. 7 provides the results of an ELISA study to confirm depletion of IGF-1 from pooled PRP.

[0017] FIG. 8A-B demonstrates that Dupuytren MFs treated with IGF-depleted PRP and ASCs show reduced lipid-filled cells. Dupuytren MFs were left untreated, co-culturedAtorney docket No. 00058-095W01with ASCs and PRP (1%) as a positive control, treated with ASCs + IGF-1 (100 ng / mL), or treated with ASCs and IGF-1 -depleted PRP (1%). (A) The cells were subjected to BODIPY staining and imaged at magnification * 10. Representative images are shown. The scale bar represents 50 pm. (B) The staining was quantified using ImageJ. The data are presented as the mean ± SD. *P < .05; **P < .01.

[0018] FIG. 9 shows IGF-1 receptor inhibition prevents Dupuytren MF transformation into lipid-filled cells. Dupuytren MFs were left untreated, cocultured with ASCs + PRP (1%), treated with ASCs + IGF-1 (100 ng / mL), or treated with an IGF-1 receptor inhibitor in the combination with ASCs + PRP or ASCs + IGF-1. The cells were fixed, stained with Oil Red O, and imaged at magnification x 10. Representative images are shown. The scale bar represents 50 pm.

[0019] FIG. 10 demonstrates that IGF-1 and hepatocyte grow th factor (HGF) induce lipid-filled cell accumulation. Duyputren MFs were untreated, treated with IGF-1 (100 ng / mL) and HGF (10 ng / mL) in a transwell format. Oil Red O staining was used to identify lipid-laden cells.

[0020] FIG. 11 demonstrates that Dihexa and IGF-1 induce lipid-filled cell accumulation. Duyputren MFs were untreated, treated with Dihexa (100 ng / mL) alone IGF-I + Dihexa (both 100 ng / mL) in trans well format. Oil Red O staining was used to identify lipid-laden cells. The lower panels represent the boxed area from the upper panels.

[0021] FIG. 12 demonstrates that ONO 1301 and IGF-I induce lipid-filled cell accumulation. Duyputren ’s myofibroblasts were untreated, treated with ONO 1301 (1 pM) alone IGF-I+ONO-1301 (100 ng / mL and 1 pM) in a transwell format. Oil Red O staining w as used to identify' lipid-laden cells. The lower panels represent the boxed area from the upper panels.

[0022] FIG. 13 demonstrates adiponectin secretion from myofibroblasts after small molecule treatment.

[0023] FIG. 14 demonstrates that the optimal concentration of Dihexa and ONO 1301 in various combinations and alone, were 50 pM.

[0024] FIG. 15 demonstrates that ONO 1301 (50 pM) on its own was most effective at inducing lipid droplets in Dupuytren MFs.

[0025] FIG. 16 show s that gene expression of C ollagen I and a-SMA w ere both significantly downregulated, while the gene expression of Collagen III was reduced in Dupuytren MFs that were treated with ONO 1301.Atorney docket No. 00058-095W01

[0026] FIG. 17 presents that gene expression of PPARG and Perilipin 1 and 2 were upregulated in Dupuytren’s myofibroblasts after treatment with ONO 1301.

[0027] FIG. 18 demonstrates that treatment with ONO 1301 induced lipid-laden cells in morphea-derived myofibroblasts.

[0028] FIG. 19 shows that Dupuytren MFs when treated with (Z)-ONO 1301 induced lipid droplet formation.DETAILED DESCRIPTION

[0029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although many methods and reagents are similar or equivalent to those described herein, the exemplary methods and materials are disclosed herein.

[0030] All publications mentioned herein are incorporated by reference in full for the purpose of describing and disclosing methodologies that might be used in connection with the description herein. The publications are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior disclosure. Moreover, with respect to any term that is presented in one or more publications that is similar to, or identical with, a term that has been expressly defined in this disclosure, the definition of the term as expressly provided in this disclosure will control in all respects.

[0031] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a fibrotic disorder" includes a plurality of such fibrotic disorders and reference to "the pro-adipogenic factor" includes reference to one or more pro-adipogenic factors and equivalents thereof known to those skilled in the art, and so forth.

[0032] Also, the use of “or” means “and / or” unless stated otherwise. Similarly, "comprise," "comprises," "comprising," "include," "includes," and "including" are interchangeable and not intended to be limiting.

[0033] It is to be further understood that where descriptions of various embodiments use the term "comprising," those skilled in the art would understand that in some specific instances, an embodiment can be alternatively described using language "consisting essentially of' or "consisting of."

[0034] The term "about," as used herein, is intended to quality' the numerical values which it modifies, denoting such a value as variable within a margin of error. When noAtorney docket No. 00058-095W01particular margin of error, such as a standard deviation to a mean value given in a chart or table of data, is recited, the term “about” should be understood to mean that range which would encompass the recited value and the range which would be included by rounding up or down to that figure as well, taking into account significant figures.

[0035] The term " adipocyte-like cells" as used herein refers to cells that are adipocytes, or cells that have many of the properties of adipocyte cells including, but not limited to, containing lipid droplets, expressing adipocyte markers like PPARG, Perilipin 1, Perilipin 2, and secreting adiponectin.

[0036] The term "disorder" as used herein is intended to be generally synonymous, and is used interchangeably with, the terms "disease," "syndrome," and "condition" (as in medical condition), in that all reflect an abnormal condition of the human or animal body or of one of its parts that impairs normal functioning, is typically manifested by distinguishing signs and symptoms.

[0037] The term "HGF -related agent" refers to compounds or small molecules, that are either analogs of HGF, truncated versions of HGF, peptides derived from HGF or are compounds or peptides that have activity that is similar to HGF. Specific examples of HGF -related agents include, but are not limited to, PG-001, CM1021, HGF alternative peptides, ONO 1301, and dihexa. In a particular embodiment, the HGF-related agent is ONO 1301.

[0038] The term " IGF- 1 -related agent" refers to compounds or small molecules that are either analogs of IGF-1, truncated versions of IGF-1, peptides derived from IGF-1 or are compounds or peptides that have activity similar to IGF-1. Specific examples of IGF-1 -related agents include, but are not limited to, des-(l-3) IGF-1 (des-IGF-1), glycine-proline-glutamate (GPE), and / or cyclic glycine-proline (cGP). In a particular embodiment, the IGF-1 -related agent is cGP.

[0039] The term "pharmaceutically acceptable" as used herein, refers to a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each component should be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of a pharmaceutical formulation. It should also be suitable for use in contact with the tissue or organ of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit / risk ratio. Examples of "pharmaceutically acceptable carriers" and "pharmaceutically acceptable excipients" can be found in the following, Remington: The Science and Practice ofAtorney docket No. 00058-095W01Pharmacy, 21st Edition; Lippincott Williams & Wilkins: Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients, 5th Edition; Rowe et al., Eds., The Pharmaceutical Press and the American Pharmaceutical Association: 2005; and Handbook of Pharmaceutical Additives, 3rd Edition; Ash and Ash Eds., Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, Gibson Ed., CRC Press LLC: Boca Raton, Fla., 2004.

[0040] The term "pro-adipogenic factor" as used herein refers to molecules or compounds that promote the transformation or reprogramming of fibroblasts or myofibroblasts into adipocyte-like cells. In a particular embodiment, the "pro-adipogenic factor" is a small molecule compound, a peptoid or a peptide. In a certain embodiment, the pro-adipogenic factor transforms or reprograms myofibroblasts, including myofibroblasts from a fibrotic disease, disorder or condition into adipocyte-like cells. In a further embodiment, fibroblasts or myofibroblasts that are contacted with a pro-adipogenic factor disclosed herein have lower expression of a-SMA, collagen type I and collagen type III, than fibroblasts or myofibroblasts not contacted with the pro-adipogenic factor.

[0001] The term "subject" as used herein, refers to an animal, including, but not limited to, a primate (e.g, human, monkey, chimpanzee, gorilla, and the like), rodents (e.g, rats, mice, gerbils, hamsters, ferrets, and the like), lagomorphs, swine (e , pig, miniature pig), equine, canine, feline, and the like. The terms "subject" and "patient" are used interchangeably herein. For example, a mammalian subject can refer to a human patient.

[0042] The term “therapeutically effective amount” refers to the amount of a compound that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disease, disorder or condition being treated. The term “therapeutically effective amount” also refers to the amount of a compound that is sufficient to elicit the biological or medical response of a cell, tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor, or clinician.

[0043] The terms "treat," "treating," and "treatment," as used herein, refers to ameliorating symptoms associated with a disease or disorder (e.g., fibrosis), including preventing or delaying the onset of the disease or disorder symptoms, and / or lessening the severity or frequency of symptoms of the disease or disorder.

[0044] Fibrotic diseases and tumor-associated fibrosis constitute a worldwide health problem that, together, are responsible for enormous morbidity and mortality. Approximately 1 in 8 people worldwide suffer from fibrotic-related diseases. Fibrotic diseases encompass a broad spectrum of clinical disorders, including systemic sclerosis, idiopathic pulmonaryAtorney docket No. 00058-095W01fibrosis, macular degeneration, chronic kidney disease, fibrosis of the liver, liver cirrhosis, and cardiac fibrosis. Similar to fibrotic diseases originating following tissue injury, fibrosis also occurs with tumor formation in pancreatic cancer and hepatocellular carcinoma.

[0005] Despite the remarkable difference in clinical manifestations and diseasecausing mechanisms, these disorders share a similar uncontrolled and progressive accumulation of fibrotic tissue in affected organs, causing their dysfunction and failure.

[0006] Recent evidence confirms that extracellular matrix stiffening plays a significant role in the initiation and progression of fibrosis. Matrix stiffness increases considerably during tissue fibrosis. For example, the stiffness of the skin, lung and liver increases from 0.5 to 1 kPa in homeostasis up to 25-100 kPa in experimental models of fibrosis. Moreover, matrix stiffening in response to tissue injury or tumours promotes mechano-activation of myofibroblasts (activated fibroblasts), which are responsible for replacing normal tissue with non-functional fibrotic tissue.

[0047] Across organs, pathological fibrosis follows similar dynamics: aberrant fibroblast activation, excessive extracellular matrix (ECM) deposition, and inadequate tissue repair. This process involves multiple cellular and molecular players that generate a fibrotic niche, characterized by a complex network of cell-to-cell and cell-to-ECM interactions.

[0048] Pathological fibrosis can result in any organ, including skin. Cutaneous (skin) fibrosis can lead to a multitude of pathologies. While these pathologies can have quite different etiologies, they all entail the supraphysiological accumulation of the ECM, fibroblasts, or their progenitors. Nearly all cutaneous fibrosis diseases, disorders and conditions can be categorized into one of four major etiological families — abnormal matrix or fibroblast accumulation, neoplasia, injury -induced fibrosis, and immune dysregulation grouped under primary or secondary etiological classes.

[0049] Primary cutaneous fibroses originate from intrinsic abnormalities in the ECM, fibroblasts, or their progenitors. The majority are caused by genetic mutations. Many of them are rare and, except for Stiff skin syndrome (SSS), can affect multiple organ systems such as the musculoskeletal system and the viscera. Abnormal matrix characterizes a group of disorders with ECM structure or turnover defects (SSS, Winchester syndrome, and hyaline fibromatosis syndrome), nuclear lamina anomalies (laminopathies), or nuclear envelope protein dysfunction (Buschke-Ollendorff syndrome). Other examples of cutaneous fibrosis resulting from abnormal matrix or fibroblast accumulation include, but are not limited to,Atorney docket No. 00058-095W01Hutchinson-Gilford progeria syndrome, restrictive dermopathy, and atypical Wemer syndrome.

[0050] Neoplastic fibroses emerge directly from the hyperproliferation of fibroblasts or their mesenchymal progenitors [dermatofibrosarcoma protuberans (DFSP), desmoid tumor, fibrous hamartoma of infancy, infantile myofibromatosis, sarcomas, and other fibrohistiocytic neoplasms] or indirectly from the transforming effects of oncogenes in the dermis (sclerotic fibroma from phosphatase and tensin homolog mutations in Cowden syndrome and connective tissue nevi from AKT serine / threonine kinase 1 mutations in Proteus syndrome). Other examples of cutaneous fibrosis resulting from neoplasia include, but are not limited to, dermatofibroma, and other fibrohistiocytic / fibroblastic / myofibroblastic tumors and sarcomas.

[0051] Secondarily, cutaneous fibroses can stem from physical, chemical, and radiation injuries or from an overactive immune system, all of which stimulate fibroblast activity and ECM production. Injury-induced cutaneous fibroses are the most heterogeneous given the countless sources of injury. Even trauma or local inflammation, for example, in acne, can lead to differential scarring — physiologic (flat, atrophic), hypertrophic, or keloidal — depending on body location and patient characteristics like ethnicity. Examples of cutaneous fibrosis resulting from injury7-induced fibrosis include, but are not limited to, bums, physiologic scar, hypertrophic scar, keloidal scar, radiation-induced fibrosis, nephrogenic systemic fibrosis, knuckle pads, lipodermatosclerosis, and dermatofibroma. Within the family of cutaneous fibrosis associated with widespread immune dysregulation, scleroderma, a progressive fibrosing disorder of multiple organs including the skin, lungs, and kidneys, is the most intensely studied due to its morbidity and mortality7. Conditions like scleromyxedema and scleredema also present with scleroderma-like rigid skin. Although both are associated with plasma cell dyscrasias, marked improvement of scleromyxedema with immunomodulatory therapy more strongly implicates the immune system in its pathogenesis than in scleredema, for which other triggers (diabetes and infections) suggest a more complex, less well-defined etiology. Despite multitudes of fibrosing disorders, there are only a handful of pathways toward fibrosis, and they all involve the activity of fibroblasts, immune cells, or the enveloping matrix. Other examples of cutaneous fibrosis resulting from immune dysregulation include, but are not limited to, lichen sclerosus, morphea, scleroderma, sclerodermatous GVHD, eosinophilic-myalgia syndrome, toxic oil syndrome, acrodermatitis chronica atrophicans, acne keloidalis nuchae, and dermatofibroma.Atorney docket No. 00058-095W01

[0052] Fibrosis can also affect non-organ tissues like connective tissue or fascia. Fascia is a dense fibrous sheath that surrounds and interpenetrates the muscles, bones, nen es, and blood vessels, binding all these structures together into a firm, compact mass. It is composed of two to three layers of dense connective tissue (collagen type I, collagen type III, and elastin fibers), and several layers of loose connective tissue (adipose tissue, glycosaminoglycan (GAG), and hyaluronic acid (HA)). It also forms continuous kinetic chains that cross multiple joints and muscle groups. This helps it store and transmit forces for more efficient movement. Fascia fibrosis is the thickening, stiffening, and scarring of connective tissue caused by excessive collagen deposition, often resulting from chronic inflammation, trauma, surgery, or aging. It reduces tissue elasticity and impairs the sliding capacity between fascial layers, leading to chronic pain, reduced mobility, and myofascial dysfunction. Examples of fascia fibrosis diseases, disorders, and conditions, include but are not limited to, plantar fibroma, Dupuytren's disease, adhesive capsulitis, Peyronie's disease, eosinophilic fasciitis, scleroderma, and chronic myofascial pain syndrome.

[0053] Dupuytren’s disease is the most common heritable disease affecting connective tissues. As many as 10 million Americans have clinical disease. Dupuytren’s contracture is characterized by extreme fibrotic changes to the fascia in the palm, which cause irreversible flexion of the fingers and substantial functional impairment for patients. Dysregulated dermal fibroblasts contribute to the development of fibroproliferative diseases, such as Dupuytren’s disease, and the resultant fascial contractures. The fibrotic microenvironment induces dermal fibroblasts to become myofibroblasts through factors, such as TGFP-1. These myofibroblasts produce disorganized collagen fibers. Moreover, alphasmooth muscle actin (a-SMA) expression by these resident myofibroblasts contributes to the persistent contracture within the palmar fascia.

[0054] Surgical treatments encompassing a variety of approaches, percutaneous release, and FDA-approved collagenase Clostridium histolyticum (CCH) are all standard treatments for Dupuytren’s contracture. In Dupuytren's disease, there is an increase in total collagen with an increased ratio of type III to I collagen in the diseased tissue. Collagenase Clostridium histolyticum, as a nonsurgical treatment, decreases collagen by enzymatically breaking it down. It also decreases a-smooth muscle actin (a-SMA) levels in myofibroblasts (MFs) in vitro, but its effect is temporary.

[0055] Dupuytren’s contracture affects patients' quality of life, makes them more self-conscious, and interferes with daily activities. Because there is no cure and recurrence isAtorney docket No. 00058-095W01unavoidable despite a multitude of surgical options, investigations into interventions with greater durability of results are highly desired. CCH was developed as a therapeutic to achieve a more minimally invasive approach to treatment. A CCH injection into the cord, followed by a manipulation procedure 24 to 72 hours later to rupture the cord, is an effective and widely adopted treatment offered as an alternative to surgery. However, patients still find this treatment painful and there can be wounds, which must be left to heal secondarily, thus limiting the minimally invasive nature of the procedure. In addition, the result is not always a complete correction of the Dupuytren’s contracture, and there is recurrence at a rate higher than many operative interventions.

[0056] Previously, tumor necrosis factor (TNF) was identified as a target to treat Dupuytren’s contracture, as it was shown to induce fibroblasts to become MFs. Thus, a clinical trial was established in the UK to determine whether anti-TNF therapy (adalimumab) altered the expression of a-SMA and thus affected collagen deposition in Dupuytren’s.Adalimumab downregulated the myofibroblast phenotype as demonstrated by reduced a-SMA and type I procollagen at 2 weeks. These data led to a phase 2b clinical trial assessing the efficacy of an intranodular injection of adalimumab in patients with early-stage Dupuytren's contracture. The results of this trial revealed that this evidence-based approach decreased nodule hardness and size. However, patient follow-up over 10 years is necessary to determine the efficacy of the anti-TNF treatment.

[0057] Anti-fibrotic treatments typically aim to target excess extracellular matrix (ECM) deposition. However, fibrosis is also associated with the loss or reduction of lipid-laden cells, a process known as lipodystrophy. Loss of these cells critically impacts tissue function and integrity, especially in the skin. Understanding the mechanisms and implications of fibrotic lipodystrophy is crucial when considering therapeutic strategies in fibrosis.Adipose-derived stem cells (ASCs), as a source of mesenchymal stem cells, are postulated to be a means of decreasing fibrosis. Adipose tissue is an excellent source for obtaining many ASCs (tens of thousands per milliliters of lipoaspirate) using liposuction, a minimally invasive procedure. In addition, the therapeutic potential of ASCs for antifibrosis is enhanced when the ASCs are cultured with platelet-rich plasma (PRP), a concentration of platelets from whole blood containing numerous antifibrotic and pro-adipogenic growth factors.

[0058] In addition, autologous fat grafting (lipoaspirate tissue) has been used to augment treatment for Dupuytren’s disease by replenishing subdermal fat loss and limitingAtorney docket No. 00058-095W01myofibroblast proliferation. Often the terminology regarding the use of ASC therapy and fat grafting in the clinical setting is incorrectly interchanged. A review focused on differentiating between fat grafting and ASC therapy with respect to the indications, harvesting, processing, application techniques, outcomes, and complications addressed this topic. Autologous fat grafting is advantageous for reconstructive and cosmetic procedures, while pure ASCs are an optimal source of cells for regenerative medicine such as soft tissue therapy for irradiated, scarred, or chronic wounds. However, no studies have examined purified homogenous ASCs to treat Dupuytren’s disease in the clinic. From an in vitro perspective, ASCs decrease contractile myofibroblasts derived from Dupuytren’s disease tissue. Fat grafting has mainly been used in addition to percutaneous aponeurotomy. These studies show varying degrees of success, and there is little consistency in the method of lipoaspirate collection.

[0059] In previous pilot studies, it was demonstrated that PRP and ASCs provide factors that induce MFs derived from Dupuytren tissue to transform into adipocyte-like lipid-laden cells in vitro. In addition, this treatment reduced fibrosis markers in a rodent model. However, adipocyte-like cells were not detected in vivo under the treatment regime in the pilot studies. Preliminary studies presented herein were to develop a treatment regime to determine whether the adipocyte-like cells generated in vitro could also be generated in vivo. The components of PRP were examined, to determine if any of these factors resulted in the production of lipid-filled adipocyte-like cells in the presence of the ASC secretome as a replacement for PRP. Based upon the foregoing studies, insulin-like growth factor 1 (IGF-1) was selected as a candidate for further research, and additional studies were performed with IGF-1 to demonstrate whether IGF-1 had similar lipogenic effects to PRP.

[0060] Antifibrotic treatments typically aim to target excess extracellular matrix (ECM) deposition. However, fibrosis is also associated with the loss or reduction of lipid-laden cells, a process known as lipodystrophy. Loss of these cells critically affect tissue function and integrity, especially in the skin. Understanding the mechanisms and implications of fibrotic lipodystrophy is important when considering therapeutic strategies in fibrosis.

[0061] As shown herein, ASCs + PRP can be used as a treatment for Dupuytren disease in vivo, as demonstrated by a rebalancing of collagen type I and collagen type III in rodent paws implanted with Dupuytren MFs. Furthermore, it was show n that ASCs + PRP transformed Dupuytren MFs into lipid-filled adipocyte-like cells. ASCs + PRP treatment again in a rodent model were tested herein with altered treatment schedules and incubation times. The rodent paws implanted with Dupuytren MFs and treated with ASCs + PRPAtorney docket No. 00058-095W01demonstrated a rebalance in collagen type I and collagen type III, reduction in a-SMA, and induction of adipocytes. Emerging evidence indicates that lipid-fdled cells secrete fibro-protective factors in tissues, and their loss during fibrotic transformation contributes to tissue dysfunction. The replenishment of fat cells within these paws after ASCs + PRP occurred after all treatment time points but was significant after two treatments. Collagenase Clostridium histolyticum (CCH) treatment did not induce adipocyte formation, which is expected as CCH targets collagen, not myofibroblasts. These findings suggest that an injection of ASCs + PRP might reduce the fibrotic condition and replenish lipid-fdled cells, decreasing the fibrotic condition. From a translational perspective, PRP has limitations, however.

[0062] The PRP used in these studies was pooled from 10 donors. In the clinic, this would likely be an autologous treatment. Thus, it was important to determine which factor in PRP contributed to adipocyte-like cell conversion in the presence of ASCs. In the studies presented herein, IGF-1 was found to function like PRP to generate adipocyte-like cells from Dupuytren MFs cocultured with ASCs. IGF-1 is a viable candidate because it induces adipogenesis in vivo by recruiting endogenous cells. Finding a factor from PRP that replaces its function in vitro is a means to alleviate the translational limitations of PRP utilization and offers a target for future mechanistic studies.

[0063] Depleting IGF-1 from PRP revealed a reduction in the formation of lipid-filled cells when Dupuytren MFs were cocultured with ASCs. This slight production of lipid-laden cells by the depleted PRP suggests that another PRP component may also contribute to this process or that the small amount of IGF-1 that remained contributed to the process. Two IGF-1 receptor inhibitors reduced the ability of IGF-1 and ASCs to transform Dupuytren MFs into lipid-laden cells to a greater degree than IGF-1 -depleted PRP. These findings suggest that Dupuytren MFs use IGF-1 receptor-mediated signaling to generate lipid-laden cells.Interestingly, IGF-1 is not capable of doing this alone. Co-culturing with ASCs was needed. Thus, synergy occurs with IGF-1 and an ASC-secreted factor that induces this process.

[0064] Thus, identifying afactor from the ASC secretome works with IGF-1 to transform Dupuytren MFs into adipocyte-like cells would yield a cell-free therapy.Furthermore, this strategy could serve to reduce the need for moderate or radical fasciectomy and limit the surgery7to a percutaneous contracture release and biologic injection aimed at reducing fibrosis, replenishing subdermal fat loss, and reducing recurrence.Attorney docket No. 00058-095W01

[0065] Dermal fat has largely been overlooked in fibrosis research. It is crucial to healthy skin, and its lipid depletion in a fibrotic context is likely to have a profound impact on skin health. The current study offers mechanisms for discovering new therapeutic targets for treating skin fibrosis, using Dupuytren disease as a model. However, research directed at fibrotic fat loss in the skin may also have implications for other organs that exhibit fibrotic lipodystrophy, such as the lungs and liver. Targeting Dupuytren’s disease by converting myofibroblasts into adipocyte-like cells offers a strategy that would reduce the need for moderate or radical fasci ectomy and limit the surgery to a percutaneous contracture release and cell-free biologic injection, aimed at reducing fibrosis and replenishing subdermal fat loss and reducing recurrence. An injectable therapeutic small molecule targeted technology' is novel to this space. A topical preparation is also possible.

[0066] It was found in the studies presented herein that fibrotic diseases, disorders, or conditions (e.g., Dupuytren's disease and Morphea) could be effectively treated by administering one or more pro-adipogenic factors that reprogram fibroblasts or myofibroblasts into adipocyte-like cells. While the studies presented herein were directed to fibrosis in fascia or skin, it should be noted that pathological fibrosis follow similar dynamics, irrespective of the tissue type or organ type: aberrant fibroblast activation, excessive extracellular matrix (ECM) deposition, and inadequate tissue repair. As the compositions and methods of the disclosure address the underlying causes for pathological fibrosis, the compositions and methods can be used to address any number of fibrotic diseases, disorders, or conditions known in the art. For example, the compositions and methods of the disclosure can be used to treat fibrotic diseases, disorders or conditions in various organs, such as in skin, lung, liver, heart and kidney. Moreover, the studies presented herein demonstrate the effectiveness of the composition and methods of the disclosure for treating fibrosis in fascia or connective tissue. Accordingly, in a particular embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that reprogram fibroblasts or myofibroblasts into adipocyte-like cells.

[0067] In the studies presented herein it was found that ONO 1301 is pro-adipogenic factor in transforming or reprogramming myofibroblasts to adipocyte-like cells. The result of which is surprising in that ONO 1301 is generally recognized in the art as being a prostacyclin agonist with thromboxane-synthase inhibitory activity. ONO 1301 wasAtorney docket No. 00058-095W01developed as a new type of PGI2 / IP receptor agonist lacking the typical prostanoid structures, including a five-membered ring and allylic alcohol. ONO 1301 has the structure of:O y Osoluble in organic solvents like dimethyl sulfoxide (DMSO). ONO 1301 is chemically and biologically stable. In mice, it has been shown that peroral administration of ONO-1301 increased hepatic gene expression and protein levels of HGF. As such. ONO 1301 for the purposes of this disclosure is classified as a HGF related compound in addition to being a pro-adipogenic factor. In the studies presented herein ONO 1301 was found to be the most effective compound in inducing lipid droplets in Dupuytren MFs. Moreover, ONO 1301 also induced lipid-laden adipocyte-like cells from morphea-derived myofibroblasts. ONO 1301 was also found to significantly suppress gene expression of Collagen type I and a-SMA and reduce expression of Collagen type III in Dupuytren MFs. Sustained release formulations of ONO 1301 (SR ONO 1301) have also been developed where ONO 1301 has been polymerized with poly (D,L-lactic co-gly colic acid). A single subcutaneous administration of SR ONO 1301 maintains effective circulating levels of ONO 1301 for 7 days, while a single subcutaneous administration of ONO 1301 is barely detectable at day 7.

[0068] In a particular embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that transform or reprogram fibroblasts or myofibroblasts into adipocyte-like cells, wherein at least one of the pro-adipogenic factors is ONO 1301 or SR ONO-131. In a further embodiment, the pharmaceutically acceptable composition comprises ONO 1301 or SR ONO-31 at a concentration selected from 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 11 pM, 12 pM, 13 pM, 14 pM, 15 pM, 16 pM, 17 pM, 18 pM, 19 pM, 20 pM, 21 pM, 22 pM, 23 pM, 24 pM, 25 pM, 26 pM, 27 pM, 28 pM, 29 pM, 30 pM, 31 pM, 32 pM, 33 pM, 34 pM, 35 pM, 36 pM, 37 pM, 38 pM, 39 pM, 40 pM. 41 pM, 42 pM, 43 pM, 44 pM, 45 pM. 46 pM. 47 pM, 48 pM, 49 pM, 50 pM, 51 pM, 52 pM, 53 pM, 54 pM. 55 pM. 56 pM, 57 pM, 58 pM, 59 pM, 60 pM, 61 pM, 62 pM, 63 pM, 64 pM, 65 pM, 66 pM, 67 pM, 68 pM, 69 pM, 70 pM, 71 pM, 72 pM, 73 pM, 74 pM, 75 pM, 76 pM, 77 pM, 78 pM, 79Atorney docket No. 00058-095W01pM, 80 pM, 81 pM, 82 pM, 83 pM, 84 pM, 85 pM, 86 pM, 87 pM, 88 pM, 89 pM, 90 pM, 91 pM. 92 pM. 93 pM, 94 pM, 95 pM, 96 pM, 97 pM, 98 pM, 99 pM, 100 pM, 110 pM, 120 pM, 130 pM, 140 pM, 150 pM, 160 pM, 170 pM, 180 pM, 190 pM, 200 pM, 250 pM, 300 pM, 350 pM, 400 pM, 450 pM, 500 pM, or a range that includes or is between any two of the foregoing concentrations (e.g., 10 pM to 100 pM, 40 pM to 70 pM, etc.), including fractional increments thereof. In another embodiment, the pharmaceutically acceptable composition provides a dose of ONO 1301 selected from 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg. 34 mg / kg, 35 mg / kg. 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 51 mg / kg, 52 mg / kg, 53 mg / kg, 54 mg / kg, 55 mg / kg, 56 mg / kg, 57 mg / kg, 58 mg / kg, 59 mg / kg, 60 mg / kg, 61 mg / kg, 62 mg / kg, 63 mg / kg, 64 mg / kg, 65 mg / kg, 66 mg / kg, 67 mg / kg. 68 mg / kg, 69 mg / kg. 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg, 100 mg / kg, 110 mg / kg, 120 mg / kg, 130 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg, 170 mg / kg, 180 mg / kg, 190 mg / kg, 200 mg / kg, 300 mg / kg, 400 mg / kg, 500 mg / kg, or a range that includes or is between any two of the foregoing concentrations (e.g., 4 mg / kg to 50 mg / kg, 10 mg / kg to 30 mg / kg, etc.), including fractional increments thereof.

[0069] An even more surprising finding presented herein is that the inactive geometric isomer of ONO 1301 , the Z isomer of ONO 1301 ((Z)-ONO 1301) was also found to also be a pro-adipogenic factor. (Z)-ONO 1301 is generally only used as an experimental control for ONO 1301, as it is recognized in the art as being inactive, i.e., lacks prostacyclin agonizing activity and lacks thromboxane-synthase inhibitory activity(e.g., see invivochem.com / product / V61065). (Z)-ONO 1301 has the structure of:. The disclosure also provides for a sustained release formulation of (Z)-ONO 1301 (SR (Z)-ONO 1301) where (Z)-ONO 1301 has beenAtorney docket No. 00058-095W01polymerized with poly (D,L-lactic co-glycolic acid) in the same manner as ONO 1301 to make SR ONO 1301.

[0070] In a particular embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that reprogram fibroblasts or myofibroblasts into adipocyte-like cells, wherein at least one of the pro-adipogenic factors is (Z)-ONO 1301, or SR (Z)-ONO 1301. In afurther embodiment, the pharmaceutically acceptable composition comprises (Z)-ONO 1301 or SR (Z)-ONO 1301 at a concentration selected from 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 11 pM, 12 pM, 13 pM, 14 pM, 15 pM. 16 pM. 17 pM. 18 pM, 19 pM, 20 pM, 21 pM, 22 pM, 23 pM, 24 pM, 25 pM, 26 pM, 27 pM, 28 pM, 29 pM, 30 pM, 31 pM, 32 pM, 33 pM, 34 pM, 35 pM, 36 pM, 37 pM, 38 pM, 39 pM, 40 pM, 41 pM, 42 pM, 43 pM, 44 pM, 45 pM, 46 pM, 47 pM, 48 pM, 49 pM, 50 pM, 51 pM, 52 pM, 53 pM, 54 pM, 55 pM, 56 pM, 57 pM, 58 pM, 59 pM, 60 pM, 61 pM, 62 pM, 63 pM. 64 pM, 65 pM, 66 pM, 67 pM, 68 pM, 69 pM, 70 pM, 71 pM, 72 pM, 73 pM, 74 pM, 75 pM, 76 pM. 77 pM. 78 pM. 79 pM, 80 pM, 81 pM, 82 pM, 83 pM, 84 pM, 85 pM, 86 pM, 87 pM, 88 pM, 89 pM, 90 pM, 91 pM, 92 pM, 93 pM, 94 pM, 95 pM, 96 pM, 97 pM, 98 pM, 99 pM, 100 pM, 110 pM, 120 pM, 130 pM, 140 pM, 150 pM, 160 pM, 170 pM. 180 pM, 190 pM, 200 pM, 250 pM. 300 pM, 350 pM, 400 pM. 450 pM, 500 pM, or a range that includes or is between any two of the foregoing concentrations (e.g., 10 pM to 100 pM, 40 pM to 70 pM, etc.) including fractional increments thereof. In another embodiment, the pharmaceutically acceptable composition provides a dose of (Z)-ONO 1301 selected from 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg. 11 mg / kg, 12 mg / kg. 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg. 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 51 mg / kg, 52 mg / kg, 53 mg / kg, 54 mg / kg, 55 mg / kg, 56 mg / kg, 57 mg / kg, 58 mg / kg, 59 mg / kg, 60 mg / kg, 61 mg / kg, 62 mg / kg, 63 mg / kg, 64 mg / kg, 65 mg / kg, 66 mg / kg, 67 mg / kg, 68 mg / kg, 69 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg, 100 mg / kg, 110 mg / kg, 120 mg / kg, 130 mg / kg, 140 mg / kg, 150 mg / kg. 160 mg / kg, 170 mg / kg, 180 mg / kg, 190 mg / kg. 200 mg / kg. 300 mg / kg, 400Atorney docket No. 00058-095W01mg / kg, 500 mg / kg, or a range that includes or is between any two of the foregoing concentrations e.g., 4 mg / kg to 50 mg / kg, 10 mg / kg to 30 mg / kg, etc.), including fractional increments thereof.

[0071] In a further embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition that is associated with fascia fibrosis in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that reprogram fibroblasts or myofibroblasts into adipocytes. Examples of fibrotic diseases, disorders or conditions that are associated with fascia fibrosis include, but are not limited to, plantar fibroma, Dupuytren's disease, adhesive capsulitis, Peyronie's disease, eosinophilic fasciitis, scleroderma, and chronic myofascial pain syndrome. In another embodiment, the fibrotic disease, disorder or condition associated with fascia fibrosis is Dupuytren's disease.

[0072] In another embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition that is associated with cutaneous (skin) fibrosis in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that transform or reprogram fibroblasts or myofibroblasts into adipocytelike cells. In a further embodiment, the cutaneous fibrosis results from abnormal matrix or fibroblast accumulation, neoplasia, injury -induced fibrosis, or immune dysregulation.Examples of fibrotic diseases, disorders or conditions that are associated with skin or cutaneous fibrosis resulting from abnormal matrix or fibroblast accumulation include, but are not limited to, stiff skin syndrome, Winchester syndrome, hyaline fibromatosis syndrome, Buschke-Ollendorf syndrome, Hutchinson-Gilford progeria syndrome, restrictive dermopathy, and atypical Wemer syndrome. Examples of fibrotic diseases, disorders or conditions that are associated with skin or cutaneous fibrosis resulting from neoplasia include, but are not limited to, dermatofibrosarcoma protuberans, desmoid tumor, fibrous hamartoma of infancy, infantile myofibromatosis, Proteus syndrome, sclerotic fibroma, dermatofibroma, and other fibrohistiocytic / fibroblastic / myofibroblastic tumors and sarcomas. Examples of fibrotic diseases, disorders or conditions that are associated with skin or cutaneous fibrosis resulting from injury-induced fibrosis include, but are not limited to, bums, physiologic scar, hypertrophic scar, keloidal scar, radiation-induced fibrosis, nephrogenic systemic fibrosis, knuckle pads, lipodermatosclerosis, and dermatofibroma.Attorney docket No. 00058-095W01Examples of fibrotic diseases, disorders or conditions that are associated with cutaneous fibrosis resulting from immune dysregulation include, but are not limited to, lichen sclerosus, morphea, scleroderma, scleredema, scleromyxedema, sclerodermatous GVHD, eosinophilic-myalgia syndrome, toxic oil syndrome, acrodermatitis chronica atrophicans, acne keloidalis nuchae, and dermatofibroma.

[0073] In another embodiment, the disclosure provides a method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising: administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that transform or reprogram fibroblasts or myofibroblasts into adipocyte-like cells, wherein the fibrotic disease, disorder or condition affects an organ. Examples of organs include, but are not limited to skin, lung, liver, heart and kidney. Examples of fibrotic diseases, disorders or conditions that affect specific organs, include the skin or cutaneous diseases, disorders or conditions mentioned above, and also include, but are not limited to, idiopathic pulmonary fibrosis, fibrosis of the liver, liver cirrhosis, cardiac fibrosis, and renal fibrosis.

[0074] For the methods of treatment presented herein, the subject in need of treatment thereof is typically a human patient. However, the methods of treatment presented herein are not limited to human subjects, but can also be applied to non-human subjects, including other mammalian subjects, like cats, dogs, horses, rats, mice, etc.

[0075] The disclosure further provides for use of pro-adipogenic factors disclosed herein for the manufacture of a medicament for the treatment of a fibrotic disease, disorder or condition in a subject in need of treatment thereof. In further embodiments, the fibrotic disease, disorder, or condition is a fibrotic disease, disorder, or condition disclosed herein. In a particular embodiment, the disclosure provides for the use of pro-adipogenic factors disclosed herein for the manufacture of a medicament for the treatment of Dupuytren's disease in a subject in need of treatment thereof. In another embodiment, the disclosure provides for the use of ONO1301, (Z)-ONO 1301, dihexa, and / or cyclic gly cine-proline (cGP) for the manufacture of a medicament for the treatment of a fibrotic disease, disorder or condition in a subject in need of treatment thereof. In yet another embodiment, the disclosure provides for the use of ONO1301 or (Z)-ONO 1301 for the manufacture of a medicament for the treatment of a fibrotic disease, disorder or condition in a subject in need of treatment thereof. In yet a further embodiment, the disclosure provides for the use of ONO1301 orAtorney docket No. 00058-095W01(Z)-ONO 1301 for the manufacture of a medicament for the treatment of Dupuytren's disease in a subject in need of treatment thereof.

[0076] The disclosure also provides for pro-adipogenic factors disclosed herein for use in the treatment of a fibrotic disease, disorder or condition in a subject in need of treatment thereof. In further embodiments, the fibrotic disease, disorder, or condition is a fibrotic disease, disorder, or condition disclosed herein. In a particular embodiment, the disclosure provides for pro-adipogenic factors disclosed herein for use in the treatment of Dupuytren's disease in a subject in need of treatment thereof. In another embodiment, the disclosure provides ONO1301, (Z)-ONO 1301, dihexa, and / or cyclic glycine-proline (cGP) for use in the treatment of a fibrotic disease, disorder or condition in a subject in need of treatment thereof. In yet another embodiment, the disclosure provides ONO 1301 or (Z)-ONO 1301 for use in the treatment of a fibrotic disease, disorder or condition in a subject in need of treatment thereof. In yet a further embodiment, the disclosure provides for ONO1301 or (Z)-ONO 1301 for use in the treatment of Dupuytren's disease in a subject in need of treatment thereof.

[0077] The disclosure further provides for a pharmaceutically acceptable composition comprising one or more pro-adiopgenic factors disclosed herein. A pharmaceutically acceptable composition comprising one or more pro-adipogenic factors disclosed herein can be in a form suitable for administration to a subject using carriers, excipients, and additives or auxiliaries. Frequently used carriers or auxiliaries include magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, milk protein, gelatin, starch, vitamins, cellulose and its derivatives, animal and vegetable oils, polyethylene glycols and solvents, such as sterile water, buffered saline, alcohols, glycerol, and polyhydric alcohols.Intravenous vehicles include fluid and nutrient replenishers. Preservatives include antimicrobial, chelating agents, and inert gases. Other pharmaceutically acceptable carriers include aqueous solutions, non-toxic excipients, including salts, preservatives, buffers and the like, as described, for instance, in Remington's Pharmaceutical Sciences, 15th ed., Easton: Mack Publishing Co., 1405-1412, 1461-1487 (1975), and The National Formulary XIV.. 14th ed., Washington: American Pharmaceutical Association (1975), the contents of which are hereby incorporated by reference. The pH and exact concentration of the various components of the pharmaceutical composition are adjusted according to routine skills in the art. See Goodman and Gilman's, The Pharmacological Basis for Therapeutics (7th ed.).Atorney docket No. 00058-095W01

[0078] The disclosure further provides for a pharmaceutically acceptable composition comprising one or more pro-adipogenic factors disclosed herein that is administered by oral administration, by injection, by topical application, or by transdermal application. Depending on the route of administration, the pharmaceutical composition can be coated with a material to protect the pharmaceutical composition from the action of enzymes, acids, and other natural conditions that may inactivate the pharmaceutical composition. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.

[0079] Pharmaceutically acceptable compositions suitable for inj ectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the composition should be sterile and should be fluid to the extent that easy syringability exists. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size, in the case of dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be typical to include isotonic agents, for example, sugars, polyalcohols, such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0080] Sterile inj ectable solutions can be prepared by incorporating the pharmaceutical composition in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, follow ed by filtered sterilization. Generally, dispersions are prepared by incorporating the pharmaceutical composition into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above.

[0081] In a particular embodiment, a pharmaceutically acceptable composition comprising one or more pro-adipogenic factors disclosed herein is administered by a microneedle transdermal patch. The delivery of therapeutical molecules through the skin,Atorney docket No. 00058-095W01particularly to its deeper layers, is impaired due to the stratum comeum layer, which acts as a barrier to foreign substances. Thus, for the past years, scientists have focused on the development of more efficient methods to deliver molecules to skin distinct layers.Microneedles, as anew class of biomedical devices, consist of an array of microscale needles. This particular biomedical device has been drawing attention due to its ability to breach the stratum comeum, forming micro-conduits to facilitate the passage of therapeutical molecules. The microneedle device has several advantages over conventional methods, such as better medication adherence, easiness, and painless self-administration. Through the formation of micron-level pores on the skin’s surface, microneedles enable the penetration of macromolecular drugs into the stratum comeum in a minimally invasive manner, increasing penetration efficiency and facilitating the direct delivery of drug molecules to the dermis. A microneedle device can also provide a rapid or long-term controlled release of drugs as a result of the rapid or slow degradation of the microneedle tip.

[0082] Thus, a “pharmaceutically acceptable carrier” is intended to include solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the pharmaceutical composition, use thereof in the therapeutic compositions and methods of treatment is contemplated. Supplementary active compounds can also be incorporated into the compositions.

[0083] It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. “Dosage unit form” as used herein, refers to physically discrete units suited as unitary dosages for the individual to be treated; each unit containing a predetermined quantity of pharmaceutical composition is calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifics for the dosage unit forms of the disclosure are related to the characteristics of the pharmaceutical composition and the particular therapeutic effect to be achieve.

[0084] The principal pharmaceutical composition is compounded for convenient and effective administration in effective amounts with a suitable pharmaceutically acceptable carrier in an acceptable dosage unit. In the case of compositions containing supplementary' active ingredients, the dosages are determined by reference to the usual dose and manner of administration of the said ingredients.Atorney docket No. 00058-095W01

[0085] Kits and articles of manufacture are also described herein. Such kits can comprise a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers can be formed from a variety of materials such as glass or plastic.

[0086] For example, the container(s) can comprise one or more pro-adipogenic factors disclosed herein, optionally in a composition or in combination with additional antifibrotic treatments as disclosed herein. The container(s) typically are made to exclude or limit light exposure to the contents of the container. Such kits optionally comprise one or more pro-adipogenic factors disclosed herein with an identifying description or label or instructions relating to its use in the methods described herein.

[0087] A kit will ty pically comprise one or more additional containers, each with one or more of various materials (such as reagents, optionally in concentrated form, and / or devices) desirable from a commercial and user standpoint for use of pro-adipogenic factors described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents; carrier, package, container, vial and / or tube labels listing contents and / or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.

[0088] A label can be on or associated with the container. A label can be on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself; a label can be associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. A label can be used to indicate that the contents are to be used for a specific analytical application. The label can also indicate directions for use of the contents, such as in the methods described herein.

[0089] The disclosure further provides that the methods and compositions described herein can be further defined by the following aspects (aspects 1 to 50):1. A method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising:administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that reprogram or transform fibroblasts or myofibroblasts into adipocyte-like cells.Attorney docket No. 00058-095W012. The method of aspect 1, wherein the fibrotic disease, disorder or condition is associated with fascia fibrosis.3. The method of aspect 1 or aspect 2, wherein the fibrotic disease, disorder or condition is selected from plantar fibroma, Dupuytren's disease, adhesive capsulitis, Peyronie's disease, eosinophilic fasciitis, scleroderma, and chronic myofascial pain syndrome.4. The method of any one of aspects 1 to 3, wherein the fibrotic disease, disorder or condition is Dupuytren's disease.5. The method of aspect 1, wherein the fibrotic disease, disorder or condition is associated with cutaneous fibrosis.6. The method of aspect 5, wherein the cutaneous fibrosis results from abnormal matrix or fibroblast accumulation, neoplasia, injury-induced fibrosis, or immune dysregulation.7. The method of aspect 5 or aspect 6, wherein the fibrotic disease, disorder or condition results from abnormal matrix or fibroblast accumulation in skin and is selected from stiff skin syndrome, Winchester syndrome, hyaline fibromatosis syndrome, Buschke-Ollendorf syndrome, Hutchinson-Gilford progeria syndrome, restrictive dermopathy, and aty pical Wemer syndrome.8. The method of aspect 5 or aspect 6, wherein the fibrotic disease, disorder or condition results from neoplasia in skin and is selected from dermatofibrosarcoma protuberans, desmoid tumor, fibrous hamartoma of infancy, infantile myofibromatosis, Proteus syndrome, sclerotic fibroma, dermatofibroma, and other fibrohistiocytic / fibroblastic / myofibroblastic tumors and sarcomas.9. The method of aspect 5 or aspect 6, wherein the fibrotic disease, disorder or condition results from injury -induced fibrosis in skin and is selected from bum, physiologic scar, hypertrophic scar, keloidal scar, radiation-induced fibrosis, nephrogenic systemic fibrosis, knuckle pads, lipodermatosclerosis, and dermatofibroma.10. The method of aspect 5 or aspect 6, wherein the fibrotic disease, disorder or condition results from immune dysregulation in skin and is selected from lichen sclerosus, morphea, scleroderma, scleredema, scleromyxedema, sclerodermatous GVHD, eosinophilic-myalgia syndrome, toxic oil syndrome, acrodermatitis chronica atrophicans, acne keloidalis nuchae, and dermatofibroma.Atorney docket No. 00058-095W0111. The method of aspect 1, wherein the fibrotic disease or condition is associated with fibrosis in an organ selected from lung, liver, heart and kidney.12. The method of aspect 11, wherein the fibrotic diseases or condition is selected from idiopathic pulmonary fibrosis, fibrosis of the liver, liver cirrhosis, cardiac fibrosis, and renal fibrosis.13. The method of any one of aspects 1 to 12, wherein the subject is a human patient.14. The method of any one of aspects 1 to 12, wherein the subject is a non-human subject.15. The method of any one of aspects 1 to 14, wherein the pharmaceutically acceptable composition further comprises a pharmaceutically acceptable diluent, excipient and / or carrier.16. The method of any one of aspects 1 to 15, wherein the pharmaceutically acceptable composition is formulated for oral administration.17. The method of any one of aspects 1 to 15, wherein the pharmaceutically acceptable composition is formulated for subcutaneous administration.18. The method of any one of aspects 1 to 15, wherein the pharmaceutically acceptable composition is formulated for topical administration.19. The method of any one of aspects 1 to 15, wherein the pharmaceutically acceptable composition is formulated for administration by a microneedle patch.20. The method of any one of aspects 1 to 19, wherein the pharmaceutically acceptable composition further comprises one or more anti-fibrotic medications.21. The method of aspect 20, wherein the one or more anti-fibrotic treatments are selected from nintedanib, pirfenidone, verapamil, collagenase, corticosteroids,my cophenolate mofetil, methotrexate, rituximab, ruxolitinib, belumosudil, celastrol, 5-fluorouracil, autologous fat grafting, tocilizumab, and triamcinolone.22. The method of any one of aspects 1 to 21, wherein the method further comprises administering to the subject one or more anti-fibrotic treatments.23. The method of aspect 22, wherein the one or more anti-fibrotic treatments are administered concurrently or sequentially with the pharmaceutically acceptable composition.24. The method of aspect 22 or aspect 23, wherein the one or more anti -fibrotic treatments are selected from nintedanib, pirfenidone, verapamil, collagenase, corticosteroids,Atorney docket No. 00058-095W01my cophenolate mofetil, methotrexate, rituximab, ruxolitinib, belumosudil, celastrol, 5-fluorouracil, autologous fat grafting, tocilizumab, and triamcinolone.25. The method of any one of aspects 1 to 24, wherein at least one of the pro-adipogenic factors is ONO 1301 having the structure of:26. The method of aspect 25. wherein the pharmaceutically acceptable composition comprises ONO 1301 at a concentration from 10 pM to 100 pM.27. The method of aspect 25 or aspect 26, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a concentration from 40 pM to 70 pM.28. The method of aspect 25, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a dose of 4 mg / kg to 50 mg / kg.29. The method of aspect 28, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a dose of 10 mg / kg to 30 mg / kg.30. The method of any one of aspects 25 to 29, wherein ONO 1301 is polymerized with poly-lactic co-glycolic acid (PLGA) microspheres to provide sustained or extended release of ONO 1301 when administered.31. The method of any one of aspects 1 to 30, wherein at least one of the pro-adipogenic factors is (Z)-ONO 1301 having the structure of:O32. The method of aspect 31, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a concentration from 10 pM to 100 pM.33. The method of aspect 31 or aspect 32, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a concentration from 40 pM to 70 pM.Atorney docket No. 00058-095W0134. The method of aspect 31 or aspect 32, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a dose of 5 mg / kg to 200 mg / kg.35. The method of aspect 31, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a dose of 10 mg / kg to 50 mg / kg.36. The method of any one of aspects 31 to 35, wherein (Z)-ONO 1301 is polymerized with poly-lactic co-gly colic acid (PLGA) microspheres to provide sustained or extended release of (Z)-ONO 1301 when administered.37. The method of any one of aspects 1 to 36, wherein the one or more pro-adipogenic factors comprises at least Insulin-like Grow th Factor 1 (IGF-1) or an IGF-1-related agent, and at least hepatocyte growth factor (HGF) or an HGF-related agent.38. The method of any one of aspects 1 to 37, wherein the one or more pro-adipogenic factors comprises IGF-1 and HGF.39. The method of any one of aspects 1 to 38, wherein the one or more pro-adipogenic factors comprises at least an IGF-l-related agent, and at least an HGF-related agent.40. The method of aspect 39, wherein the at least one IGF-l-related agent comprises des-(l-3) IGF-I (des-IGF-I), glycine-proline-glutamate (GPE), and / or cyclic glycine-proline (cGP).41. The method of aspect 40. wherein the at least one IGF-l-related agent comprises cGP.42. The method of aspect 39, wherein the at least one HGF-related agent comprises ONO-1301 and / or dihexa having the structure of:OH43. The method of any one of aspects 1 to 42, wherein the one or more pro-adipogenic factors comprises cGP, dihexa, and ONO- 1301.44. The method of any one of aspects 1 to 4, wherein the method is used in combination with one or more Dupuytren disease treatments.Atorney docket No. 00058-095W0145. The method of aspect 44, wherein the Dupuytren disease treatments is selected from collagenase injections or needle aponeurotomy, surgical fasciotomy, steroid injections, radiation therapy, and fasciectomy.46. A method to reprogram or transform fibroblasts or myofibroblasts into adipocytes, comprising:contacting fibroblasts or myofibroblasts with a composition comprising ONO-1301 having the structure of:or (Z)-ONO 1301 having the structure of:OQ47. The method of aspect 46, wherein ONO-1301 or (Z)-ONO 1301 is used at a concentration from 10 pM to 100 pM.48. The method of aspect 46 or aspect 47, wherein ONO-1301 or (Z)-ONO 1301 is used at a concentration from 40 pM to 70 pM.49. The method of any one of aspects 46 to 48, wherein the fibroblasts or myofibroblasts are contacted in vitro.50. The method of any one of aspects 46 to 48, wherein the fibroblasts or myofibroblasts are contacted in vivo.51. The method of any one of aspects 46 to 50, wherein the fibroblasts or myofibroblasts are myofibroblasts of a fibrotic disease, disorder or condition.52. The method of any one of aspect 51, wherein the myofibroblasts are Dupuytren myofibroblasts.

[0090] The following examples are intended to illustrate but not limit the disclosure. While they are ty pical of those that might be used, other procedures known to those skilled in the art may alternatively be used.Atorney docket No. 00058-095W01EXAMPLES

[0091] Fibroblast isolation and culture. After institutional review board approval, the palmar fascia from discarded surgical tissue collected at the University of California Irvine Medical Center was used to isolate Dupuytren myofibroblasts (MFs). The normal human dermal fibroblasts (NHDFs) were purchased from ATCC. The Dupuytren tissue was trimmed to remove excess fat tissue, soaked in dispase (5 U / mL; Stem Cell Technologies), washed in enzyme-free cell dissociation buffer (Millipore), and then in HEPES-buffered saline (Sigma- Aldrich). The tissue was digested using type II collagenase (Gibco) and halted with 10% fetal bovine serum (FBS; Gibco). The digested tissue was filtered through a cell strainer (Fisher), centrifuged, and resuspended in Minimum Essential Media (Gibco) containing 10% FBS and 1% penicillin and streptomycin (P / S; Coming). The cells were grown to confluence and split using trypsin (Gibco). The cells used for the experiments were from passages 4-8.

[0092] Characterization of MFs. After the MFs were isolated, the expression of a-SMA was assessed. Normal human dermal fibroblasts were plated, left untreated, or treated with TGF-pi (15 ng / mL for 48 hours) to induce MF differentiation, and Dupuytren MFs ere plated simultaneously without treatment. Immunofluorescence was performed by fixing the cells and using a primary antibody to detect a-SMA (Cell Signaling Technologies) and a fluorescently conjugated secondary' antibody (Alexa Fluor 488, Invitrogen). The nuclei were counter-stained with DAPI (Millipore). Fluorescence was quantified using ImageJ. The percent of a-SMA-positive cells were NHDF 7.6% (±2.1), NHDF ± TGF- 1 29.8% (±4.4), and Dupuytren MFs 43.3% (±9.9), verifying the isolation of MFs from the Dupuytren disease tissue.

[0093] ASC isolation and culture. The lipoaspirate was obtained from healthy individuals undergoing elective liposuction at the University of California Irvine Medical Center. The tissue was w ashed in phosphate-buffered saline and digested with type I collagenase I (Stem Cell Technologies). Dulbecco’s modified Eagle medium (Coming) supplemented with 10% FBS and 1% P / S was used to stop the digestion. The solution was filtered through a cell strainer (Fisher), centrifuged, and the cell pellet was resuspended in Dulbecco’s modified Eagle medium with 10% FBS and 1% P / S. The ASCs were characterized as described in Banyard et al., J Cell Mol Med. 19(1): 21 e30 (2015). The cells were used in passages 4-8.Atorney docket No. 00058-095W01

[0094] Dupuytren animal model. The isolated Dupuytren MFs and NHDFs were implanted (8 x io6cells / paw) into the left and right forepaws of 27 male nude rats (Rowett Nude mu / mu athymic)) respectively, to establish a Dupuytren disease animal model under Institutional Animal Care and Use Committee approval. The rats were randomly allocated to three groups 2 months after implanting the cells. Based on a pilot study, power analysis revealed that two rats are needed to achieve at least 95% power to test using a two-sided paired t test at a 2.5% significance level. Bonferroni correction was applied to account for multiple group comparisons. Three animals were selected per group to account for any issues with the experiment. The forepaws were treated with 100 rnL of saline or a mix of PRP (1%) + ASCs (1 x 106cells / paw in 100 mL), or CCH (0.58 mg / mL in 100 mL) to compare the clinical treatment (n = 9 / group). Previous in vitro data revealed that lipid-filled cells were only induced when ASCs and PRP were combined. Therefore, these components were not assessed alone in the rats. The forepaw tissue was harvested after 1 week for nine rats (n = 3 / group). The 18 remaining rats received the treatments described above. After another week, the forepaw tissue was harvested from nine rats (n = 3 / group), and the nine remaining rats received the last treatments. Finally, 1 week later, the forepaw tissue from the final nine rats was harvested. The harvested tissues were fixed in formalin and paraffin-embedded following standard protocols. After sectioning, Masson’s trichrome staining (Polysciences) was performed to assess collagen (e.g.. see procedures outlined in FIG. 1).

[0095] Immunohistochemistry (IHC) was performed to evaluate a-SMA. collagen type I, collagen type III, and perilipin expressions. Briefly, the sections were blocked in hydrogen peroxide after deparaffinization and rehydration. Antigen retrieval was performed using a trypsin enzy matic kit (Abeam). A Mouse and Rabbit-Specific HRP / DAB (ABC) Detection IHC kit (Abeam) was used, and the primary antibodies were a-SMA (1 : 100; Cell Signaling), Collagen I alpha 1 (1:100; Novus Biologicals), Collagen III alpha 1 (1: 100;Novus Biologicals), and perilipin (1:100; Cell Signaling). The slides were mounted and imaged at 4. Five fields of view' per slide w ere used to quantify the protein expression (Image J [NIH]).

[0096] Immunohistochemistry images were deconvoluted to obtain the DAB-positive area. The mean gray value w as used to determine the signal intensity of each image. Intensity units were then converted to optical density by using the following formula: optical densify = log(maximum intensity / mean intensify), where maximum intensify = 255. Adipocyte count and area were calculated using the Adipocyte Tools plugin. Adipocytes were segmentedAtorney docket No. 00058-095W01using the percentile thresholding method with a minimum adipocyte size of 100 and a maximum size of 200,000.

[0097] Cell culture and treatments. Dupuytren MFs were seeded (0.5 * 105cells / well) into 24-well plates. The cells were treated at approximately 80% confluency (~1.9 x 105cells / well). Untreated Dupuytren MFs served as a negative control. Dupuytren MFs cocultured with ASCs (transwell culture system; ASCs were added at 1 x 105cells / insert) and treated with PRP ( 1 %) served as a positive control, as determined in Ziegler et al. , J Hand SurgAm. 48(9):914e922 (2023). The PRP was acquired from healthy donors collected at the University of California, Irvine using an Eclipse PRP (3.2 platelet multiplier and leukocyte poor; Eclipse). The PRP (10 donors) was pooled to account for individual variability. For the experimental treatments. Dupuytren MFs were treated with recombinant human IGF-I (Irvine Scientific; 10 ng / mL or 100 ng / mL) alone or with the ASCs coculture. To inhibit IGF, NVPAEW541 (MedChemExpress), an inhibitor of the IGF-I receptor (IGF-1R), or BMS-536924 (Med-ChemExpress), a competitive and selective IGF-1R inhibitor, was used at 10 mM to treat the Dupuytren MFs. The cells were assessed at 4 weeks using Oil Red O (MilliporeSigma) or BODIPY (ThermoFisher) to reveal lipid droplets. The cells were imaged and analyzed using ImageJ (e g., see procedures outlined in FIG. 1).

[0098] IGF depletion from PRP and validation. Pooled PRP was rotated with an IGF-I antibody (2.5 mg; Peprotech) or an isotype control antibody overnight at 4 °C. Then, protein A / G beads (SellekChem) were added to the PRP, which was rotated for 20 minutes at 4 °C. The beads were collected using a magnet, and the depleted PRP was collected. An ELISA (Ray BioTech) was conducted following the manufacturer’s instructions to verify the depletion of IGF from the PRP.

[0099] Small molecule treatment of Duputren myofibroblasts. Dupuytren MFs were untreated or treated with HGF (50 ng / mL) + IGF (100 ng / mL), IGF-I + Dihexa (both 100 ng / mL), IGF + ONO 1301 (both 100 ng / mL), cGP (100 nM) + HGF (50 ng / mL), or cGP (100 nM) + Dihexa (100 ng / mL) + ONO 1301 (100 ng / mL). Treatments were applied every 3-4 days, and the supernatant was collected after 2 weeks of treatment. ELISA detected the secretion of adiponectin. (n=2)

[0100] Statistical analysis. All the quantitative data are expressed as the mean ± SEM. To assess the differences between the groups, an unpaired, two-tailed t test was performed. A value ofP < .05 was considered statistically significant.Atorney docket No. 00058-095W01

[0101] Validation of the Dupuytren rodent model. Paws implanted with Dupuytren MFs demonstrated more collagen accumulation than those implanted with NHDFs, as shown by the Masson’s Trichrome staining (e.g., see FIG. 2A). The Dupuytren MF paws had significantly (P < .05) more collagen type I (see Fig. 2A-B) and higher collagen type III expression (not significant; P > .05; e.g., see FIG. 2A, C) than those implanted with NHDFs. Furthermore, the type III to I collagen ratio was higher in the Dupuytren MF paws (e.g., see FIG. 2D). Finally, the expression of a-SMA within the tissue (not associated with the blood vessels) was significantly greater in the Dupuytren MF paws (P < .05; e.g., see FIG. 2A, E).These increased disease markers in the paws implanted with Dupuytren MFs compared to those with NHDFs validated the original data.

[0102] Fibrotic marker reduction with ASCs + PRP treatment. Masson' s trichrome staining was similar across all time points for the Dupuytren MF paws (a representative image is shown in FIG. 3A). After the first two treatments with CCH or ASCs + PRP, there was a reduction in overall collagen. How ever, after the third treatment with ASCs + PRP, the reduction was greater than the CCH-treated paws (e.g, see FIG. 3A). Collagenase Clostridium histolyticum and ASCs + PRP reduced collagen type I expression after the first treatment. However, after the second treatment, collagen type I expression increased with CCH but declined with ASCs + PRP compared with the control (not significant; P > .5; e.g., see FIG. 3B). After one treatment, CCH and ASCs + PRP reduced collagen type III expression. The difference between the reduced levels of the treatment groups was significant (P < .01); the ASCs + PRP treatment demonstrated a more substantial reduction. For the second treatment, the decrease w as significant only for the ASCs + PRP treated paw s. After the third treatment, collagen type III expression w as unchanged with CCH and lower with ASCs + PRP (P > .05; see FIG.3C). After three treatments, the collagen III / I ratio was 1.4 for the Dupuytren MF group, 1.9 for the CCH treatment, and 1.0 for the ASCs + PRP treatment (see FIG. 3D). Thus, the overall decrease in collagen w as likely because of the reduction of collagen ty pe III.

[0103] The expression of a-SMA was the greatest in the paws implanted with Dupuytren MFs at the tw o-treatment time point, likely reflecting the highest activity of the MFs. Collagenase Clostridium histolyticum and ASCs + PRP significantly inhibited a-SMA expression (45% [P < .05] and 64% [P < .01], respectively; e.g., see FIG. 3E). However, the inhibition was greater for ASCs + PRP. Thus, treating rodent paws with ASCs + PRP effectively reduced collagen and a-SMA.Atorney docket No. 00058-095W01

[0104] Emergence of adipocytes with ASCs + PRP treatment. The adipocyte counts for the NHDF paws were significantly greater (P < .01) than the Dupuytren MFs at the 2-week treatment time. Collagenase Clostridium histolyticum treatment did not change the adipocyte counts in the Dupuytren MF paws. In contrast, ASCs + PRP treatment led to a significant increase (P < .01) in adipocyte counts, reaching a number similar to the NHDF paws. After one and three treatments, the adipocyte count was the greatest for the ASCs + PRP group (e.g., see FIG.4).

[0105] IGF-I replaces PRP transforming DMFs into lipid-filled cells in the presence of ASCs. Insulin-like grow th factor 1 (10 ng / mL) demonstrated lipogenic effects similar to PRP (ASCs + PRP:87% lipid-laden cells and ASCs + IGF-1:83% lipid-laden cells; e.g., see FIG. 5). Insulin-like growth factor 1 was then used at increasing concentrations to optimize the transformation, and 100 ng / mL was selected (e.g, see FIG.6).

[0106] Next, IGF-l was depleted from the pooled PRP, and the depletion was confirmed by ELISA (P < .01; e.g., see FIG. 7). When Dupuytren MFs were cocultured with IGF- 1 -depleted PRP and ASCs, lipid-laden cells were reduced by 64%. In contrast, the cells treated with ASCs + PRP and ASCs + IGF-1 showed significant induction of lipid-filled cells (P < .05 and P < .01, respectively; e.g., see FIG. 8A-B).

[0107] Finally, treating Dupuytren MFs with IGF-1 receptor inhibitor (NVP-AEW541) blocked the induction of lipid-filled cells when Dupuytren MFs were treated with ASCs + PRP and ASCs + IGF-1. ASCs + PRP showed 84% lipid-laden cells, and IGF-1 receptor inhibition reduced it to 13%. ASCs + IGF-1 showed 80% lipid-laden cells, and IGF-1 receptor inhibition lowered it to 11% (e.g. , see FIG. 9). Similar results were demonstrated with BMS-536924, another IGF-1 receptor inhibitor.

[0108] Evaluating growth factors and other small molecules that work in a synergistic manner in converting myofibroblasts into adipocytes. It was recognized that stem cell transplantation therapy has limitations, including potential tumorigenicity, cell rejection, emboli formation, undesired differentiation, and infection transmission. Cell-free-based therapies avoid these issues. Thus, examining the ASC secretome, a set of complex soluble factors, including cytokines, chemokines, growth factors, and vesicles, appears to offer therapeutic promise. Working in a paracrine fashion, ASC-secreted factors have been demonstrated to target lung, liver, and cardiac fibrosis and systemic sclerosis.

[0109] It was found herein that hepatocyte growth factor (HGF), a secreted factor from ASCs, demonstrated lipogenic effects when combined with IGF-1 (e.g.. see FIG. 10).Atorney docket No. 00058-095W01Thus, IGF-1 and HGF served as adipogenic factors that could transform myofibroblasts into adipocytes. Often growth factors do not meet the clinical trial endpoint due to unfavorable pharmacological properties. However, peptide growth factor mimetics or analogs are favorable alternatives. Thus, the next phase of discovery involved a search for mimetics or analogs to replace IGF-1 and HGF.

[0110] In the circulation, unbound IGF-1 can be enzymatically metabolized to des-(l-3) IGF-1 (des-IGF-1) and glycine-proline-glutamate (GPE). GPE is enzymatically unstable and is quickly metabolized into single amino acids and dipeptides, including cyclic glycineproline (cGP). Thus, it was postulated herein that cGP could be used in place of IGF-1 to transform myofibroblasts into lipid-filled cells. When cGP was combined with HGF or with ASCs, lipid-filled cell accumulation resulted.

[0111] Next, small molecule HGF alternatives was investigated. Two small molecule compounds were identified, Dihexa and ONO 1301. Dihexa is an HGF mimetic that is a synthetically derived Angiotensin IV analog. ONO 1301 is a prostaglandin 12 agonist that increases the production of HGF in various cell types, fibroblasts. Dihexa and ONO 1301 both induced lipid-filled cell accumulation either alone or with IGF-1 (e.g., see FIG. 11 and FIG. 12, respectively).

[0112] Preliminary' experiments with cGP, Dihexa, and ONO 1301used in combination dramatically greatly induced adiponectin secretion in comparison to combinations that did not include all three agents, or agents used alone (e.g., see FIG. 13).

[0113] Optimizing cGP, Dihexa and ONO 1301 concentrations for lipid droplet formation in normal human dermal fibroblasts. The next step was exploring cGP, Dihexa, and ONO 1301 to optimize concentrations for lipid droplet formation. After several rounds, it was confirmed that cGP was not adding any boost in lipid droplet formation when cotreated with Dihexa and ONO- 1301. cGP w as not used in further experiments. Dihexa and ONO 1301 were then tested at higher concentrations (10, 50, and 100 pM) in various combinations and alone. The optimal concentration for both w as determined to be 50 pM (e.g., see FIG. 14). 100 pM concentrations of Dihexa and ONO 1301 caused cell death.

[0114] Evaluating the effect of Dihexa and ONO 1301 on Dupuytren myofibroblasts using optimized concentrations. Next, Dihexa and ONO 1301 were evaluated and compared for their abilities to induce lipid droplet in Dupuytren MFs. After multiple experiments, it was determined that ONO 1301 (50 pM) on its own was most effective at inducing lipid droplets in Dupuytren’s myofibroblasts (e.g.. see FIG. 15).Atorney docket No. 00058-095W01

[0115] Evaluating the effect of ONO 1301 on gene expression of Collagen I, Collagen II, a-SMA, PPARG, Perilipin 1 and Perilipin 2 in Dupuytren myofibroblasts.Next, Dupuytren MFs were treated with ONO 1301 (50 pM) for two weeks (4 treatments) and harvested for RNA isolation. qPCR was conducted to evaluate the expression of key genes associated with fibrosis. Collagen type I and a-SMA were both significantly downregulated, while Collagen type III was reduced (e.g., see FIG. 16). Gene expression of PPARG and Perihpin 1 and 2 were also evaluated after ONO 1301 treatment. The upregulation of Perilipin 2 revealed replenished lipid droplet formation in Dupuytren’s myofibroblasts. In addition, a subset of these cells was Perilipin 1 positive, suggesting differentiation into mature adipocytes, as supported by PPARG upregulation (e.g., see FIG.17).

[0116] Determining the effect of ONO 1301 treatment in other types of myofibroblasts. To determine whether ONO 1301 induced lipid droplet in another type of myofibroblasts, cells were isolated from biopsy tissue from a patient with morphea (a sclerosing skin disorder). Treatment with ONO 1301 induced lipid-laden cells in morphea-derived myofibroblasts (e.g., see FIG. 18).

[0117] Determining the effect of other enantiomeric forms of ONO 1301 in Dupuytren’s myofibroblasts. (Z)-ONO 1301 is described in the art as being an inactive isomer of ONO-1301 and is only used as an experimental control (see invivochem.com / product / V61065). It was surprisingly found that Dupuytren MFs. when treated with the “inactive form” of ONO 1301, (Z)-ONO 1301, induced lipid droplet formation (e.g., see FIG. 19).

[0118] A number of embodiments have been described herein. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of this disclosure. Accordingly, other embodiments are within the scope of the following claims.

Claims

Atorney docket No. 00058-095W01WHAT IS CLAIMED IS:

1. A method of treating a fibrotic disease, disorder or condition in a subject in need of treatment thereof, comprising:administering to the subject a pharmaceutically acceptable composition comprising therapeutically effective amounts of one or more pro-adipogenic factors that transform or reprogram fibroblasts or myofibroblasts into adipocyte-like cells.

2. The method of claim 1, wherein the fibrotic disease, disorder or condition is associated with fascia fibrosis.

3. The method of claim 2, wherein the fibrotic disease, disorder or condition is selected from plantar fibroma, Dupuytren's disease, adhesive capsulitis, Peyronie's disease, eosinophilic fasciitis, scleroderma, and chronic myofascial pain syndrome.

4. The method of claim 3, wherein the fibrotic disease, disorder or condition is Dupuytren's disease.

5. The method of claim 1, wherein the fibrotic disease, disorder or condition is associated with cutaneous fibrosis.

6. The method of claim 5, wherein the cutaneous fibrosis results from abnormal matrix or fibroblast accumulation, neoplasia, injury-induced fibrosis, or immune dysregulation.

7. The method of claim 6, wherein the fibrotic disease, disorder or condition results from abnormal matrix or fibroblast accumulation in skin and is selected from stiff skin syndrome, Winchester syndrome, hyaline fibromatosis syndrome, Buschke-Ollendorf syndrome, Hutchinson-Gilford progeria syndrome, restrictive dermopathy, and atypical Wemer syndrome.

8. The method of claim 6, wherein the fibrotic disease, disorder or condition results from neoplasia in skin and is selected from dermatofibrosarcoma protuberans, desmoid tumor, fibrous hamartoma of infancy, infantile myofibromatosis, Proteus syndrome, scleroticAtorney docket No. 00058-095W01fibroma, dermatofibroma, and other fibrohistiocytic / fibroblastic / myofibroblastic tumors and sarcomas.

9. The method of claim 6, wherein the fibrotic disease, disorder or condition results from inj ury-induced fibrosis in skin and is selected from bum, physiologic scar, hypertrophic scar, keloidal scar, radiation-induced fibrosis, nephrogenic systemic fibrosis, knuckle pads, lipodermatosclerosis, and dermatofibroma.

10. The method of claim 6, wherein the fibrotic disease, disorder or condition results from immune dysregulation in skin and is selected from lichen sclerosus, morphea, scleroderma, scleredema, scleromyxedema, sclerodermatous GVHD, eosinophilic-myalgia syndrome, toxic oil syndrome, acrodermatitis chronica atrophicans, acne keloidalis nuchae, and dermatofibroma.

11. The method of claim 1, wherein the fibrotic disease or condition is associated with fibrosis in an organ selected from lung, liver, heart and kidney.

12. The method of claim 11, wherein the fibrotic diseases or condition is selected from idiopathic pulmonary fibrosis, fibrosis of the liver, liver cirrhosis, cardiac fibrosis, and renal fibrosis.

13. The method of claim 1, wherein the subject is a human patient.

14. The method of claim 1, wherein the subject is a non-human subject.

15. The method of claim 1, wherein the pharmaceutically acceptable composition further comprises a pharmaceutically acceptable diluent, excipient and / or carrier.

16. The method of claim 1 , wherein the pharmaceutically acceptable composition is formulated for oral administration.

17. The method of claim 1, wherein the pharmaceutically acceptable composition is formulated for subcutaneous administration.Atorney docket No. 00058-095W0118. The method of claim 1, wherein the pharmaceutically acceptable composition is formulated for topical administration.

19. The method of claim 1, wherein the pharmaceutically acceptable composition is formulated for administration by a microneedle patch.

20. The method of claim 1, wherein the pharmaceutically acceptable composition further comprises one or more anti-fibrotic medications.

21. The method of claim 20, wherein the one or more anti-fibrotic treatments are selected from nintedanib, pirfenidone, verapamil, collagenase, corticosteroids, mycophenolate mofetil, methotrexate, rituximab, ruxolitinib, belumosudil, celastrol, 5-fluorouracil, autologous fat grafting, tocilizumab, and triamcinolone.

22. The method of claim 1, wherein the method further comprises administering to the subject one or more anti-fibrotic treatments.

23. The method of claim 22, wherein the one or more anti-fibrotic treatments are administered concurrently or sequentially with the pharmaceutically acceptable composition.

24. The method of claim 22, wherein the one or more anti-fibrotic treatments are selected from nintedanib, pirfenidone, verapamil, collagenase, corticosteroids, mycophenolate mofetil, methotrexate, rituximab, ruxolitinib. belumosudil. celastrol. 5-fluorouracil, autologous fat grafting, tocilizumab, and triamcinolone.

25. The method of any one of claims 1 to 24, wherein at least one of the pro-adipogenic factors is ONO 1301 having the structure of:Atorney docket No. 00058-095W0126. The method of claim 25, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a concentration from 10 p.M to 100 pM.

27. The method of claim 26, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a concentration from 40 pM to 70 pM.

28. The method of claim 25, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a dose of 4 mg / kg to 50 mg / kg.

29. The method of claim 27, wherein the pharmaceutically acceptable composition comprises ONO 1301 at a dose of 10 mg / kg to 30 mg / kg.

30. The method of claim 25, wherein ONO 1301 is polymerized with poly-lactic co-gly colic acid (PLGA) microspheres to provide sustained or extended release of ONO 1301 when administered.

31. The method of any one of claims 1 to 24, wherein at least one of the pro-adipogenic factors is (Z)-ONO 1301 having the structure of:

32. The method of claim 31, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a concentration from 10 pM to 100 pM.

33. The method of claim 32, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a concentration from 40 pM to 70 pM.

34. The method of claim 31, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a dose of 5 mg / kg to 200 mg / kg.Atorney docket No. 00058-095W0135. The method of claim 34, wherein the pharmaceutically acceptable composition comprises (Z)-ONO 1301 at a dose of 10 mg / kg to 50 mg / kg.

36. The method of claim 31, wherein (Z)-ONO 1301 is polymerized with poly -lactic co-glycolic acid (PLGA) microspheres to provide sustained or extended release of (Z)-ONO 1301 when administered.

37. The method of any one of claims 1 to 24, wherein the one or more pro-adipogenic factors comprises at least Insulin-like Growth Factor 1 (IGF-1) or an IGF-l-related agent, and at least hepatocyte growth factor (HGF) or an HGF-related agent.

38. The method of claim 35, wherein the one or more pro-adipogenic factors comprises IGF-1 and HGF.

39. The method of claim 37, wherein the one or more pro-adipogenic factors comprises at least an IGF-l-related agent, and at least an HGF-related agent.

40. The method of claim 39, wherein the at least one IGF-l-related agent comprises des-(1-3) IGF-I (des-IGF-I), glycine-proline-glutamate (GPE), and / or cyclic glycine-proline (cGP).

41. The method of claim 40, wherein the at least one IGF-l-related agent comprises cGP.

42. The method of claim 37, wherein the at least one HGF-related agent comprises ONO- 1301 and / or dihexa having the structure of:Atorney docket No. 00058-095W0143. The method of any one of claims 1 to 24, wherein the one or more pro-adipogenic factors comprises cGP, dihexa, and ONO-1301.

44. The method of claim 1, wherein the method is used in combination with one or more Dupuytren disease treatments.

45. The method of claim 44, wherein the Dupuytren disease treatments is selected from collagenase injections or needle aponeurotomy, surgical fasciotomy, steroid injections, radiation therapy, and fasciectomy.

46. A method to reprogram or transform fibroblasts or myofibroblasts into adipocyte-like cells, comprising:contacting fibroblasts or myofibroblasts with a composition comprising ONO-1301 having the structure ofor (Z)-ONO 1301 having the structure of:

47. The method of claim 46, wherein ONO-1301 or (Z)-ONO 1301 is used at a concentration from 10 pM to 100 pM.

48. The method of claim 47, wherein ONO-1301 or (Z)-ONO 1301 is used at a concentration from 40 pM to 70 pM.

49. The method of claim 46, wherein the fibroblasts or myofibroblasts are contacted inAttorney docket No. 00058-095W01vitro.

50. The method of claim 46, wherein the fibroblasts or myofibroblasts are contacted in vivo.

51. The method of claim 46, wherein the fibroblasts or myofibroblasts are myofibroblasts of a fibrotic disease, disorder or condition.

52. The method of claim 51, wherein the my ofibroblasts are Dupuytren myofibroblasts.

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