EA042390B1 - METHOD OF THERAPEUTIC TREATMENT OF FIBROSIS - Google Patents

Description

Technical field

Compositions and methods according to the present description generally belong to the field of treatment of fibrotic disease.

State of the art

Fibrosis is a pathogenic feature of a wide range of conditions affecting a wide variety of tissues, including the liver (examples: non-alcoholic steatohepatitis, glycogen storage disease type IX, cirrhosis), lung (examples: chronic interstitial lung disease, pneumoconiosis, silicosis, emphysema, fibrosing lung diseases , idiopathic pulmonary fibrosis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia), vascular system (examples: diffuse interstitial fibrosis; atherosclerosis), heart (examples: cardiac fibrosis; atrial fibrosis; endomyocardial fibrosis), skin (examples: keloid lesions , nephrogenic systemic fibrosis, scleroderma), joints and interstitial tissues (examples: arthrofibrosis, Dupuytren's disease), pancreas (example: pancreatitis), mouth (examples: fibrous proliferative lesions of the oral cavity), intestines (examples: fibrosing strictures, e.g. associated with Crohn's disease), the brain (glial scarring, leptomeningeal fibrosis associated with bacterial meningitis). Fibrosis can also occur as a result of negative environmental influences or various traumas, such as, for example, exposure to ionizing radiation (such as during cancer treatment), as a result of rupture of a breast cyst (causing palpable damage to breast tissue) and in general in the result of excess collagen deposition after injury or tissue damage, such as after trauma or surgery.

Although some types of fibrosis are associated with existing genetic predispositions (example: Dupuytren's disease), most types of fibrosis are associated with long-term inflammation of damaged tissue (examples: liver fibrosis and pneumoconial fibrosis). Symptoms can be as minor as itching and aesthetic problems (eg, in the case of keloid skin lesions) or as severe as lung failure and death (eg, as symptoms of end-stage pulmonary fibrosis and cardiac fibroses). Although fibrosis is essentially irreversible once established, treatments exist to slow the progression of various fibrotic conditions or to improve the course of fibrosis or fibrotic conditions. Current anti-fibrotic treatments include anti-inflammatory compounds such as pirfenidone and the fibroblast growth factor receptor antagonist nintedanib. For cutaneous and subcutaneous fibrosis, examples of current therapies include surgery, phototherapy, and injections of collagenase from the bacteria Clostridium histolyticum. However, due to the irreversible nature of various fibroses, as well as the limited efficacy of current therapies, there remains a need for additional therapeutic approaches for this class of conditions.

Brief description of the invention

Disclosed herein is a method for the therapeutic treatment of fibrosis resulting in a reduction in the amount of extracellular matrix proteins present in one or more tissues of a subject, comprising administering to a subject in need thereof one or more compounds having a structure selected from the group consisting of

or their pharmaceutically acceptable salts.

- 1 042390

According to the methods described herein, the compound to be administered may include one or more compounds having a structure selected from the group consisting of

(Compound 2), or their pharmaceutically acceptable salts.

According to the methods disclosed herein, the compounds described above may be administered for the treatment, amelioration, prevention, or cure of one or more fibrotic conditions selected from glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), non-alcoholic steatohepatitis (NASH), cirrhosis, hepatitis, scleroderma, alcoholic fatty liver disease, atherosclerosis, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone fibrosis brain, liver fibrosis, cirrhosis of the liver and gallbladder, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic int interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, chronic kidney disease, type II diabetes, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, fibrosis after injection, complications after surgery, fibrous chronic allograft vasculopathy and/or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, postvasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis, Dupuytren's disease, dermatomyositis-polymyositis, mixed connective tissue disease, fibrosis x proliferative lesions of the oral cavity, fibrosing intestinal strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, radiation-induced fibrosis, breast cyst rupture fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, psoriasis, or any combination of them. Methods and compositions as described herein may include primary fibrosis or a condition in which said fibrosis, fibrotic condition, or fibrotic symptom is secondary to or symptomatic of another condition.

In some embodiments of the methods disclosed herein, said fibrosis, fibrotic condition, or fibrotic symptom may include one or more of scleroderma, atherosclerosis, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial lung disease, chronic interstitial lung disease, pneumoconiosis, silicosis, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis , macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, injection fibrosis, fibrotic complications after surgery, fibrous chronic allograft vasculopathy, fibrosis associated with ischemic reperfusion injury, arthrofibrosis, Dupuytren's disease, fibrous proliferative lesions of the oral cavity, fibrosing intestinal strictures, glial scarring, leptomeningeal fibrosis, radiation-induced fibrosis, breast cyst rupture fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, or any of these combinations.

In some other embodiments, said fibrosis, fibrous condition, or fibrotic

The symptom may be secondary to one or more of glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), non-alcoholic steatohepatitis (NASH), cirrhosis, hepatitis, scleroderma, alcoholic fatty liver disease, atherosclerosis, asthma, cirrhosis of the gallbladder, diffuse parenchymal lung disease, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, emphysema, chronic kidney disease, type II diabetes, macular degeneration, chronic organ transplant rejection, postvasectomy pain syndrome, rheumatoid arthritis, dermatomyositis-polymyositis, mixed connective tissue disease, Crohn's disease, meningitis, systemic lupus erythematosus or their symptoms or sequelae, or other diseases or conditions resulting in excessive deposition of extracellular matrix components such as collagen, which may be affected by interference with the TRe pathway, or any combination of them.

In some other embodiments, said fibrosis, fibrotic condition, or fibrotic symptom may include a symptom or consequence of GSD III, GSD IX, non-alcoholic steatohepatitis, cirrhosis of the liver or pancreas, Dupuytren's disease, scleroderma, idiopathic pulmonary fibrosis, or alcoholic fatty liver disease, or any combination thereof. .

The methods of the present disclosure may also include a method of treating fibrosis, a fibrotic condition, or a fibrotic symptom in a subject, comprising administering one or more of

subject that needs it.

Also provided are methods according to the present disclosure for administering to a subject one or more of the compounds listed above, wherein said subject has one or more conditions selected from glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), type IX glycogen storage (GSD IX), hepatitis, scleroderma, atherosclerosis, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, gallbladder cirrhosis, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, kidney fibrosis, chronic kidney disease, type II diabetes, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, fibrosis after injection, complications after surgery, fibrous chronic allograft vasculopathy and/or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, postvasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis , Dupuytren's disease, dermatomyositis-polymyositis, mixed connective tissue disease, fibrous proliferative lesions of the oral cavity, fibrosing intestinal strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, radiation-induced fibrosis, breast cyst rupture fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, or any combination thereof. The methods of the present disclosure involve said administration, wherein said condition is primary fibrosis, secondary fibrosis, or a fibrotic symptom of the condition.

In the methods of the present disclosure, primary fibrosis may include one or more of scleroderma, atherosclerosis, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, spleen fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, idiopathic interstitial fibrosis. fibrosis, diffuse interstitial fibrosis, interstitial lung disease, chronic interstitial lung disease, pneumoconiosis, silicosis, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, macular degeneration, keloid lesions, hypertrophic scar, systemic nephrogenic fibrosis, fibrosis after injection, fibrotic complications after surgery, fibrous chronic allograft vasculopathy, fibrosis associated with ischemic reperfusion injury, arthrofibrosis, Dupuytren's disease, fib oral proliferative lesions, fibrosing intestinal strictures, glial scarring, leptomeningeal fibrosis, radiation-induced fibrosis, breast cyst rupture fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, or any combination thereof.

According to the methods of the present disclosure, secondary fibrosis may include fibrosis associated with one or more of glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), hepatitis, scleroderma , atherosclerosis, asthma, cirrhosis of the gallbladder, diffuse parenchymal lung disease, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, emphysema, chronic kidney disease, type II diabetes, macular degeneration, chronic organ transplant rejection, post-vasectomy pain syndrome, rheumatoid arthritis, dermatomyositis-polymyositis, mixed connective tissue diseases, Crohn's disease, meningitis, systemic lupus erythematosus or their symptoms or consequences, or other diseases or conditions resulting in excessive deposition of extracellular matrix components such as collagen, which may be affected by interference with the TRe pathway, or any combination thereof.

In the methods of the present disclosure, primary fibrosis may include a symptom or consequence of GSD III, GSD IX, pancreatic cirrhosis, Dupuytren's disease, scleroderma, idiopathic pulmonary fibrosis, or alcoholic fatty liver disease, or any combination thereof.

According to the methods of the present disclosure, the compositions to be administered may further comprise one or more pharmaceutically acceptable excipients and may be formulated for oral, intravenous, intra-arterial, intestinal, rectal, vaginal, nasal, pulmonary, topical, intradermal, transdermal, buccal, translingual , sublingual or ophthalmic administration, or any combination thereof.

According to the methods of the present disclosure, a subject to be administered the compositions listed above may exhibit abnormal or excessive deposition of type I, Ia, or III collagen. In some embodiments of the methods and compositions of the present disclosure, administration of said composition results in the prevention, amelioration, or cure of said fibrosis, fibrotic condition, or fibrotic symptom, and may also result in a decrease in the amount of extracellular matrix proteins present in one or more tissues of said subject. In some embodiments, said reduction in the amount of extracellular matrix proteins present in one or more tissues of said subject may include a decrease in the amount of collagen present in one or more tissues of said subject, and may also include a decrease in type I, type Ia, or type III collagen. present in one or more tissues of the specified subject.

Brief description of the drawings

In FIG. 1 shows total liver hydroxyproline in mice after 8 weeks of treatment with vehicle, low dose compound 2 (see example 1), high dose compound 2 (see example 1), compound 1 or elafibranor (control). Animals treated with compound 2 show lower levels of total hydroxyproline in the liver than animals treated with control or sham treatment.

In FIG. 2 shows representative images of the liver stained with Picro-Sirius Red (to visualize collagen I and III deposition, red spot) at the end of treatment after 8 weeks of vehicle, low dose compound 2 (see example 1), high dose compound 2 (see example 1), compound 1 or elafibranor (control) (magnification 10x, scale bar=200 µm).

In FIG. 3 shows representative images of a liver stained against type I collagen.

- 4 042390 (collal) (Southern Biotech, Cat. 131001) at the end of treatment after 8 weeks of vehicle, low dose compound 2 (see example 1), high dose compound 2 (see example 1), compound 1 or elafibranor ( control) (magnification 20x, scale bar=100 µm).

In FIG. 4 shows total liver ColA1 after biopsy after 8 weeks of vehicle, low dose compound 2 (see example 1), high dose compound 2 (see example 1), compound 1 or elafibranor (control). Compound 2 treated animals show lower total liver ColA1 levels than control or sham treated animals.

In FIG. 5 shows liver biopsy staining with Picro-Sirius Red (PSR) according to histological quantification after 8 weeks of vehicle, low dose Compound 2 (see Example 1), high dose Compound 2 (see Example 1), Compound 1 or elafibranor (control). Total (mg/liver) liver collagen 1 and 3 was determined by morphometry after staining with Picro-Sirius Red. Compound 1-treated liver sections showed lower PSR staining than liver sections from sham treated animals; liver sections of Compound 2 treated animals showed lower PSR staining than liver sections of control or sham treated animals. Data are expressed as mean ± standard error of the mean (n=11-12).

In FIG. 6 shows the percentage reduction in total hepatic triglycerides, cholesterol and lipids, and the percentage reduction in NAS for Compound 2 treated animals compared to vehicle controls after 8 weeks of Compound 2 administration, studies were performed in a diet-induced NASH model in mice (see example 6).

In FIG. 7 shows the percent reduction in hepatic fibrosis in Compound 2 treated animals compared to vehicle controls after 8 weeks of Compound 2 administration in a diet-induced NASH mouse model (see Example 6). Liver fibrosis was assessed by fibrosis score, the level of type I collagen present, and the level of hydroxyproline present in post-treatment liver samples.

In FIG. 8 shows the percentage reduction in profibrogenic gene expression in Compound 2 treated animals compared to vehicle controls after 8 weeks of Compound 2 administration, studies were performed in a diet-induced NASH mouse model (see Example 6). The profibrogenic gene expression was assessed by the expression levels of Cola1, Col3a1, aSMA and Galectin 1 in post-treatment liver samples.

Detailed description of the invention

Provided herein are compounds and methods for treating fibrosis, fibrotic conditions, or fibrotic symptoms by administering thyroid hormone β receptor (TRe) agonists. In some embodiments, such fibrosis, fibrotic conditions, or conditions causing fibrotic symptoms may include glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), non-alcoholic steatohepatitis (NASH), cirrhosis, hepatitis, scleroderma, alcoholic fatty liver disease, atherosclerosis, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, cirrhosis of the liver and gallbladder, spleen fibrosis, scleroderma pulmonary fibrosis, idiopathic pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, kidney fibrosis, chronic kidney disease, type II diabetes, macular degeneration, keloid Lesions, hypertrophic scar, nephrogenic systemic fibrosis, fibrosis after injection, complications after surgery, fibrous chronic allograft vasculopathy and/or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, postvasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis , Dupuytren's disease, dermatomyositis-polymyositis, mixed connective tissue disease, fibrous proliferative lesions of the oral cavity, fibrosing intestinal strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, radiation-induced fibrosis, breast cyst rupture fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, or symptoms or sequelae thereof, or other diseases or conditions resulting in excessive deposition of extracellular matrix components such as like collagen, which can be affected by interference in the TRe pathway. Such conditions may be associated with inflammation and/or trauma and, in addition, may be associated with responses mediated

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TGF-β-dependent pathways that can be modulated by thyroid hormones (see, for example, Alfonso-Merino et al., Proc. Nat. Acad. Sci., 113(24):E3451-60 (2016), which is included in present description because it discloses the ability of thyroid hormones to modulate transforming growth factor beta (TGF-β) signaling and associated fibrosis in mice). Because the

TGF-B dependent pathways are involved in fibroblast differentiation and stimulation of collagen production and thyroid hormones such as triiodothyronine (T3) and thyroxine (T4) can act on these pathways via the TRe receptor, compositions and methods for preventing, eliminating or reducing collagen deposition in one or more tissues of the subject by administering TRe agonist compounds.

Definitions.

The term mammal is used in its usual biological sense. Thus, it specifically includes humans and non-human mammals such as dogs, cats, horses, donkeys, mules, cows, domestic buffaloes, camels, llamas, alpacas, bison, yaks, goats, sheep, pigs, elk, deer, domestic antelopes and non-human primates, as well as many other species.

Subject as used herein means a human or non-human mammal including, but not limited to, dog, cat, horse, donkey, mule, cow, domestic buffalo, camel, llama, alpaca, bison, yak, goat, sheep, pig , elk, deer, domestic antelope or non-human primate selected for treatment or therapy.

A subject with suspected presence means a subject exhibiting one or more clinical signs of a disease or condition. In some embodiments, said disease or condition is one or more fibroses, fibrotic conditions, or fibrotic symptoms. In some embodiments, said disease or condition is scleroderma. In some embodiments, said disease or condition is non-alcoholic steatohepatitis. In some embodiments, said disease or condition is cirrhosis. In some embodiments, said disease or condition is non-alcoholic fatty liver disease. In some embodiments, said disease or condition is idiopathic pulmonary fibrosis. In some embodiments, said disease or condition is atherosclerosis. In some embodiments, said disease or condition is hepatitis, alcoholic fatty liver disease, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, cirrhosis of the liver and gallbladder, spleen fibrosis, scleroderma , pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis; interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, chronic kidney disease, type II diabetes, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, injection fibrosis, complications after surgery, fibrous chronic allograft vasculopathy and/ or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, post-vasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis, Dupuytren's disease, dermatomyositis-polymyositis, mixed connective tissue disease, fibrous proliferative lesions of the oral cavity, fibrosing bowel strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, radiation-induced fibrosis, cyst rupture fibrosis breast, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis or their symptoms or consequences, or other diseases or conditions leading to excessive deposition of extracellular matrix components.

In the context of the present description, fibrosis refers to the abnormal deposition of extracellular matrix proteins. Such proteins include, but are not limited to, collagen, elastin, fibronectin, laminin, keratin, keratin, keratin sulfate, fibrin, perlecan, agrin or agrecan. As used herein, collagen refers to any of the collagen subtypes, including, but not limited to, I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI , XVII or XVIII type. Illustrative types and subtypes of collagen specifically include type I, type Ia, type II, type III, type IV, and type V. As used herein, fibrosis may occur on its own or as a symptom or consequence of another condition. As used herein, fibrosis may result from a genetic condition, genetic predisposition, environmental stress, trauma, trauma healing, an autoimmune condition or chronic inflammation, a chronic inflammatory condition, or other condition resulting in abnormal or excessive deposition of extracellular components. matrix. Fibrosis as referred to herein can be assessed by assaying or determining the presence or amount of one or more biomarkers. Biomarkers for the presence of fibrosis include, but are not limited to, expression of the Cola1, Col3a1, aSMA and/or Galectin1 genes, or any combination or product thereof. Diagnosis or evaluation of fibrosis can also be performed by determining the presence or amount of type I collagen and/or hydroxyproline, or any combination or product thereof. Diagnosis or assessment of fibrosis can also be performed using histological, histochemical or immunohistochemical analysis of one or more samples taken from the subject.

Glycogen storage disease refers to any one or more disorders from a group of disorders characterized by dysfunction in the synthesis, transport, or utilization of glycogen, usually due to loss of activity of an essential enzyme. Glycogen storage diseases are generally classified by type according to their symptoms and etiology. Known types include GSD type 0 (aglycogenesis, glycogen synthase deficiency); GSD type 1 (Girke's disease, glucose-6-phosphatase translocase/transporter deficiency, GSD I); GSD type 2 (Pompe disease, alpha-1-4-glucosidase deficiency, GSD II); GSD type 3 (Corey's disease, Forbes' disease, limit dextrinosis, debranching enzyme disease; amyl 1-6-glucosidase deficiency due to loss of glucosidase and/or transferase activity, GSD III); GSD type 4 (Andersen's disease, glycogen phosphorylase deficiency, branching enzyme deficiency, amylopectinosis, glycogen branching enzyme deficiency; amyl-1,4-1,6-transglucosidase deficiency, GSD IV); GSD type 5 (McArdle disease; glycogen phosphorylase deficiency (muscle type), GSD V); GSD type 6 (Hers disease; glycogen phosphorylase E deficiency (liver type), GSD VI); GSD type 7 (Tarui disease; phosphofructokinase deficiency, GSD VII); GSD type 8, 9 (GSD with defects in the phosphorylase activation system; phosphorylase kinase deficiency (liver or muscle isoforms), GSD VIII and GSD IX); GSD type 10 (cyclic adenosine monophosphate (AMP)-dependent kinase deficiency, GSD X); GSD type 11 (Fanconi-Bickel syndrome; deficiency of glucose transporter type 2 (GLUT2), GSD XI) and GSD type 12 (aldolase A deficiency, GSD XII). There are also known subtypes of glycogen storage diseases, in particular GSD 1a, which result from mutations in the gene encoding glucose-6-phosphatase (G6PC), and, among other symptoms, lead to excessive accumulation of glycogen and lipids in the liver tissue, hepatomegaly, adenomas liver and hepatocellular carcinoma. Symptoms of glycogen storage diseases can include high or low blood sugar, insulin insensitivity, myopathies, and hepatic symptoms such as steatosis, hyperlipidemia, hypercholesterolemia, cardiomegaly, hepatomegaly, fibrosis, cirrhosis, hepatocellular adenoma, and hepatocellular carcinoma. Symptoms may also include insulin insensitivity, high or low blood glucose, kidney dysfunction, and/or fibrosis.

Subject in need means a subject identified as in need of therapy or treatment.

The therapeutic effect relieves to some extent one or more of the symptoms of the disease or disorder and includes curing said disease or disorder. Cure means that the symptoms of the active disease have been eliminated. However, some long-term or irreversible effects of the disease (such as extensive tissue damage) may exist even after treatment.

To treat, treat or carry out treatment in the context of the present description refers to the administration of a pharmaceutical composition for prophylactic and/or therapeutic purposes. The term prophylactic treatment refers to the treatment of a patient who does not yet have the underlying disease or disorder, but who is susceptible to or otherwise at risk for a particular disease or disorder, said treatment reducing the likelihood that the patient will develop said disease or disorder. The term therapeutic treatment refers to the administration of treatment to a patient who already has a disease or disorder.

Prevention or avoidance refers to delaying or preventing the onset, development, or progression of a condition or disease over a specified period of time, including weeks, months, or years.

An improvement in course means a decrease in the severity of at least one indicator of a condition or disease. In some embodiments, improving the course includes delaying or slowing the progression of one or more indicators of the condition or disease. The severity of the indicators can be determined using subjective or objective criteria that are known to experts in this field of technology.

Modulation means changing a function or activity. In some embodiments, modulation means an increase in gene expression. In some embodiments, modulation means a reduction in gene expression. In some embodiments, modulation means an increase or decrease in the total serum content of a particular protein. In some embodiments, modulation means an increase or decrease in serum levels of free, specific protein. In some implementations, modulation means increasing or decreasing

- 7 042390 total serum content of a specific non-protein factor. In some embodiments, modulation means an increase or decrease in serum levels of a specific free non-protein factor. In some embodiments, modulation means an increase or decrease in the overall bioavailability of a particular protein. In some embodiments, modulation means an increase or decrease in the overall bioavailability of a particular non-protein factor.

Administration means providing a pharmaceutical agent or composition to a subject and includes, but is not limited to, administration by a healthcare professional and self-administration.

Administration of the compounds disclosed herein, or their pharmaceutically acceptable salts, may be by any of the routes of administration accepted for agents of similar use, including, but not limited to, orally, subcutaneously, intravenously, intranasally, topically, transdermally, intraperitoneally, intramuscularly, intrapulmonaryly, vaginally, rectally or intraocularly. Oral and parenteral routes of administration are traditional in the treatment of indications, which are the subject of preferred embodiments.

Parenteral administration means administration by injection or infusion. Parenteral administration includes, but is not limited to, subcutaneous administration, intravenous administration, intramuscular administration, intra-arterial administration, and intracranial administration.

Subcutaneous administration means administration directly under the skin.

Intravenous administration means administration into a vein.

Intra-arterial administration means administration into an artery.

The term agent includes any substance, molecule, element, compound, structure, or combination thereof. It includes, but is not limited to, for example, a protein, a polypeptide, a peptide or mimetic, a small organic molecule, a polysaccharide, a polynucleotide, and the like. It may be a natural product, a synthetic compound, or a chemical compound, or a combination of two or more substances.

A pharmaceutical agent means a substance that provides a therapeutic effect when administered to a subject.

Pharmaceutical composition means a mixture of substances suitable for administration to an individual, which contains a pharmaceutical agent. For example, the pharmaceutical composition may contain a modified oligonucleotide and a sterile aqueous solution.

An active pharmaceutical ingredient means a substance in a pharmaceutical composition that provides the desired effect.

The term pharmaceutically acceptable salt refers to salts that retain the biological efficacy and properties of the compounds to which they are associated, and which are not biologically or otherwise undesirable. In many cases, the compounds according to the present description are able to form acidic and/or basic salts due to the presence of phenolic and/or phosphonate groups or similar groups. One skilled in the art will recognize that the degree of protonation of any or all of these compounds may vary depending on the pH and ionic characteristics of the surrounding solution, and thus the present description assumes several states of charge for each compound. Pharmaceutically acceptable acid addition salts can be prepared using inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be prepared using inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, hydroxides of sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like; particularly preferred are the ammonium, potassium, sodium, calcium and magnesium salts. Organic bases from which salts may be derived include, for example, primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, such as, in particular, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. Many such salts are known in the art as described in WO 87/05297 by Johnston et al., published September 11, 1987 (incorporated herein by reference in its entirety).

A solvate refers to a compound obtained by reacting a solvent and an EPI, a metabolite, or a salt thereof. Suitable solvates are pharmaceutically acceptable solvates, including hydrates.

The term atherosclerosis refers to a condition characterized by unevenly distributed lipid deposits in the intima of large and medium-sized arteries, where such deposits provoke fibrosis and calcification. Atherosclerosis increases the risk of angina, stroke, heart attack, or other heart or cardiovascular conditions.

Connections.

In some embodiments, TRe agonists for the use described herein include compounds according to formula I

B ⁹ B ¹ B'

Formula I wherein G is selected from the group consisting of -O-, -S-, -S(=O)-, -S(=O) ₂ -, -Se-, -CH ₂ -, -CF ₂ -, - CHF-, -C(O)-, -CH(OH)-, -CH(C ₁ -C ₄ -αkyl)-, -CH(C ₁ -C ₄ -αlkoxy)-, -C(=CH ₂ ) -, -NH- and -N(C ₁ -C ₄ -alkyl)-;

T is selected from the group consisting of -(CR ^a 2)k-, -CR ^b =CR ^b -(CR ^a 2)n-, -(CR ^a 2)n-CR ^b =CR ^b -, -(CR ^a 2)-, CR ^b =CR ^b -(CR ^a 2)-, -O(CR ^b 2)(CR ^a 2)n-, -S(CR ^b 2)(CR ^a 2)n-, N(R ^c )(CR ^b 2)(CR ^a 2)n-, N(R ^b )C(O)(CR ^a 2)n, -C(O)(CR ^a 2)m-, -(CR ^a 2) _m C(O)-, -(CR ^a 2)C(O)(CR ^a 2)n, -(CR ^a 2)nC(O)(CR ^a 2)- and -C(O)NH(CR ^b 2)(CR ^a 2)p-;

k is an integer of 1-4;

m is an integer of 0-3;

n is an integer of 0-2;

p is an integer of 0-1;

each R ^a is independently selected from the group consisting of hydrogen, optionally substituted -C1-C4-αalkyl, halogen, -OH, optionally substituted -O-C1-C4-αalkyl, -OCF3, optionally substituted -S-C1-C4-αalkyl , -NR ^b R ^c , optionally substituted -C2-C ₄ -alkenyl and optionally substituted -C ₂ -C ₄ -alkenyl; with the proviso that if one R ^a is attached to C through an O, S or N atom, then the other R ^a attached to the same C is hydrogen or attached through a carbon atom;

each R ^b is independently selected from the group consisting of hydrogen and optionally substituted -C ₁ -C ₄ -αkyl;

each R ^c is independently selected from the group consisting of hydrogen and optionally substituted -C ₁ -C ₄ -αkyl, optionally substituted -C(O)-C ₁ -C ₄ -αkyl, and -C(O)H;

each of R ¹ and R ² is independently selected from the group consisting of halogen, optionally substituted -C ₁ -C ₄ -αalkyl, optionally substituted -SC ₁ -C ₃ -αalkyl, optionally substituted -C ₂ -C ₄ -αalkenyl, optionally substituted -C ₂ -C ₄ -αkynyl, -CF ₃ , -OCF3, optionally substituted -OC ₁ -C ₃ -αkyl and cyano;

each of R ⁶ , R ⁷ , R ⁸ and R ⁹ is independently selected from the group consisting of hydrogen, halogen, optionally substituted -C C1-C4-αkyl, optionally substituted -S-C1-C3-αlkyl, optionally substituted -C2- C4-alkenyl, optionally substituted -C2-C4-αkynyl, -CF3, -OCF3, optionally substituted -O-C1-C3-αkyl and cyano; or R ⁶ and T are combined together with the carbon atoms to which they are attached to form a ring of 5-6 atoms, including 0-2 heteroatoms, independently selected from -NR ¹ -, -O- and -S-, provided that if there are 2 heteroatoms in the ring and both heteroatoms are other than nitrogen, then both heteroatoms must be separated by at least one carbon atom; and X is attached to said ring via a direct bond to a ring carbon atom or via a -(CR ^a 2)- or -C(O)- group linked to a ring carbon atom or a ring nitrogen atom;

R ¹ is selected from the group consisting of hydrogen, -C(O)C ₁ -C ₄ -αkyla, -C ₁ -C ₄ -αkyla and -C ₁ -C ₄ -aryl;

R ³ and R ⁴ are independently selected from the group consisting of hydrogen, halogen, -CF ₃ , -OCF3, cyano, optionally substituted -C ₁ -C ₁₂ -αkyl, optionally substituted -C ₂ -C ₁₂ -αalkenyl, optionally substituted - C2-C12-αkynyl, -SR ^d , -S(=O)R ^e , -S(=O)2R ^e , -S(=O)2NR ^f R ^g , -C(O)OR ^h , -C( O)R ^e , -N(R ^b )C(O)NR ^f R ^g , -N(R ^b )S(=O)2R ^e , -N(R ^b )S(=O)2NR ^f R ^g and -NR ^f R ^g ;

^d each R ^d is selected from the group consisting of optionally substituted -C1-C12-αkyl, optionally substituted -C2-C12-αalkenyl, optionally substituted -C2-C12-αkynyl, optionally substituted -(CR ^b 2) _n -aryl, optionally substituted -(CR ^b 2)n-cycloαalkyl, optionally substituted -(CR ^b 2) _n -heterocycloαalkyl, and -C(O)NR ^f R ^g ;

each Re ^is selected from the group consisting of optionally substituted -C1-C12-αkyl, optionally substituted -C2-C12-αalkenyl, optionally substituted -C2-C12-αkynyl, optionally substituted -(CR ^a 2) _n -aryl, optionally substituted -(CR ^a 2)n-cycloαalkyl and optionally substituted -(CR ^a 2) _n -heterocycloαalkyl;

each of R ^f and R ^g is independently selected from the group consisting of hydrogen, optionally substituted -C1-C12-αlkyl, optionally substituted -C2-C12-αalkenyl, optionally substituted -C2-C12-αkynyl, optionally substituted -(CR ^b 2 ) _n -aryl, optionally substituted -(CR ^b ₂ ) _n -cycloαalkyl, and optionally substituted -(CR ^b 2)n-heterocycloalkyl, or R ^f and R ^g together may form an optionally substituted heterocyclic ring which may contain a second heterogroup selected from the group consisting of O, NRC and S, wherein said optionally substituted heterocyclic ring may be substituted with 0-4 substituents selected from the group consisting of optionally substituted -C1- _C4 alkyl, -OR ^b , oxo, cyano, -CF3, optionally substituted phenyl and -C(O)OR ^h ;

each R ^h is selected from the group consisting of optionally substituted -C1-C12-alkyl, optionally substituted -C2-C12-alkenyl, optionally substituted -C2-C12-alkynyl, optionally substituted -(CR ^b 2) _n aryl, optionally substituted - (CR ^b ₂ ) _n cycloalkyl and optionally substituted -(CR ^b 2) n heterocycloalkyl;

R ⁵ is selected from the group consisting of -OH, optionally substituted -OC1-C6 alkyl, OC(O)R ^e , -OC(O)OR ^h , -F, -NHC(O)R ^e , -NHS(= O)R ^e , -NHS(=O)2R ^e , -NHC(=S)NH(R ^h ) and -NHC(O)NH(R ^h );

X is P(O)YR ¹¹ Y'R ¹¹ ;

each of Y and Y' is independently selected from the group consisting of -O- and -NRv-;

when Y and Y' are -O-, R ¹¹ attached to -O- is independently selected from the group consisting of -H, alkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, optionally substituted CH ₂ -heterocycloalkyl, wherein cyclic the fragment contains a carbonate or thiocarbonate of an optionally substituted alkylaryl, -C(R ^z )2OC(O)NR ^z 2, -NR ^z -C(O)-R ^y , -C(R ^z )2-OC(O)R ^y , -C(R ^z )2-OC(O)OR ^y , -C(R ^z )2OC(O)SR ^y , -alkyl-SC(O)R ^y , -alkyl-SS-alkylhydroxy and -alkyl-SSS -alkylhydroxy;

if Y and Y' are -NR ^v -, then R ¹¹ attached to -NR ^v - is independently selected from the group consisting of -H, -[C(R ^z b]q-COOR ^y , -C(R ^x bCOOR ^Y , -[C(R ^z )2]qC(O)SR ^y and -cycloalkylene-COOR ^y ;

when Y is -O- and Y' is NRv, then R ¹¹ attached to -O- is independently selected from the group consisting of -H, alkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, optionally substituted CH2-heterocycloalkyl, while the cyclic fragment contains a carbonate or thiocarbonate, optionally substituted -alkylaryl, -C(R ^z )2OC(O)NR ^z 2, -NR ^z -C(O)-R ^y , -C(R ^z )2-OC( O)R ^y , -C(R ^z )2-OC(O)OR ^y , -C(R ^z hOC(O)SR ^y , -alkyl-SC(O)R ^y , -alkyl-S-Salkylhydroxy and - alkyl-SSS-alkylhydroxy, and R ¹¹ attached to -NRv- is independently selected from the group consisting of H, -[C(R ^z )2]q-COOR ^y , -C(R ^x )2COOR ^y , -[ C(R ^z )2] _q -C(O)SR ^y and -cycloalkylene-COOR ^y ; or if Y and Y' are independently selected from -O- and NRv, then R ¹¹ and R ¹¹ together are -alkyl-SS -alkyl- to form a cyclic group, or R ¹¹ and R ¹¹ together represent a group

where V, W and W' are independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted aralkyl, heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, optionally substituted 1-alkenyl, and optionally substituted 1-alkynyl; or

V and Z are linked to each other through an additional 3-5 atoms to form a cyclic group containing 5-7 atoms, where 0-1 atom is a heteroatom and the remaining atoms are carbon substituted by hydroxy, acyloxy, alkylthiocarbonyloxy, alkoxycarbonyloxy, or aryloxycarbonyloxy attached to a carbon atom, i. e. three atoms at both Y-groups attached to phosphorus; or

V and Z are bonded to each other through an additional 3-5 atoms to form a cyclic group where 0-1 atom is a heteroatom and the remaining atoms are carbon, which is fused to an aryl group in the beta and gamma positions relative to the Y attached to phosphorus; or

V and W are bonded to each other via an additional 3 carbon atoms to form an optionally substituted cyclic group containing 6 carbon atoms and substituted with one substituent selected from the group consisting of hydroxy, acyloxy, alkoxycarbonyloxy, alkylthiocarbonyloxy and aryloxycarbonyloxy attached to one of the indicated atoms carbon, i.e. three atoms at Y attached to phosphorus; or

Z and W are linked to each other through an additional 3-5 atoms to form a cyclic group, where 0-1 atom is a heteroatom and the remaining atoms are carbon and V must be aryl, substituted aryl, heteroaryl or substituted heteroaryl; or

W and W' are bonded to each other through an additional 2-5 atoms to form a cyclic group, where 0-2 atoms are heteroatoms and the remaining atoms are carbon and

V must be aryl, substituted aryl, heteroaryl or substituted heteroaryl;

Z is selected from the group consisting of -CHRzOH, -CHR ^z OC(O)R ^y , -CHR ^z OC(S)R ^y , -CHR ^z OC(S)OR ^y , -CHR ^z OC(O)SR ^y , -CHR ^z OCO2R ^y , -OR ^z , -SR ^z , -CHR ^z N3, -CH ₂ -aryl, -CH(aryl)OH, -CH(CH=CR ^z ₂ )OH, -CH(C-CR ^z )OH, -R ^z , -NR ^z 2 , -OCOR ^y , -OCO2R ^y , -SCOR ^y , -SCO2R ^y , -NHCOR ^z , -NHCO2R ^y , -CH2NH-aryl, -(CH2)q-OR ^z and -(CH2)q- ^SRz ;

q is an integer of 2 or 3;

each R ^z is selected from the group consisting of R ^y and -H;

each R ^y is selected from the group consisting of alkyl, aryl, heterocycloalkyl and aralkyl;

each R ^x is independently selected from the group consisting of -H and alkyl, or R ^x and R ^x together form a cyclic alkyl group;

each Rv is selected from the group consisting of -H, lower alkyl, acyloxyalkyl, alkoxycarbonyloxyalkyl and lower acyl;

and their pharmaceutically acceptable salts.

In some embodiments, a compound of formula I satisfies the following conditions:

a) if G is -O-, T is -CH2-, R ¹ and R ² are each bromine, R ³ is isopropyl, R ⁴ is hydrogen and R ⁵ is -OH, then X is not is P(O)(OH) ₂ or P(O)(OCH ₂ CH ₃ ) ₂ ;

b) not all of V, Z, W, W' are -H; And

c) if Z is -R ^z , then at least one of V, W and W' is not -H, alkyl, aralkyl or heterocycloalkyl;

d) if G is -O-, T is -(CH ₂ ) ₁ . ₄ -, R ¹ and R ² are independently halogen, alkyl and cycloalkyl, R ³ is alkyl, R ⁴ is hydrogen, and R ⁵ is -OH, then X is not -P(O)(OH) ₂ or -P(O)(O-lower alkyl) ₂ ; And

e) if G is -O-, R ⁵ is -NHC(O)R ^e , -NHS(=O)1.2R ^e , -NHC(S)NH(R ^b ) or -NHC(O)NH( R ^h ), T is -(CH2) ^m -, -CH=CH-, -O(CH ₂ ) ₁ . ₂ - or -NH(CH ₂ ) ₁ . ₂ -, then X is not -P(O)(OH) ₂ or -P(O)(OH)NH ₂ .

In some embodiments, said compound is selected from one or more of the following compounds:

0 °Kz ϊ 1 ο ^ζ \ HO Ο Ρ ο jjx - II ¹ XI _? ^ο ^Hz ? ⁿ ? H ₃ C'''^''^O'' ^X '''T — he o H ₃ C H3C ΛΑΛΑ ⁿ ° -| _sC ' ^x x_^X. _o / _{Xp_he} o or their pharmaceutically acceptable salts. E^O (Compound 1) Oh 7 (Compound 2) (Connection 3) or (Compound 4)

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In other embodiments, said compound is selected from

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0 sn ₃ NSG H ₃ C' CFI ₃ Cl '^erV ⁴ « H ₃ C Ύ Τ T1 no h ₃ c^ ^- cn ₃ Cl ^H!C> XXYj CH ₃ ^H,c 0O00 ZhU H ₃ r CH ₃ ZSTl ⁴ NSU H ₃ C^ 41 but ^^\V° IT p II I ® SG ί\ ii ^27-2 / ^OH HO 7-7 /\ HO OH ^-Ji ⁴¹ ' ^{3 p} \ / OH o HO 1 o 24-4 P-- OH 1 OH 1 0 7-8 K^p< / ΌΗ HO

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OH or their pharmaceutically acceptable salts.

The compounds described above may be prepared according to known methods, including those described in US Pat. No. 7,829,552, which is incorporated herein by reference in its entirety. Additional TRe agonists are described in US Pat. No. 7,514,419; U.S. Patent Application Publication No. 2009/002895; U.S. Patent Application Publication No. 2010/0081634; US Patent Application Publication No. 2012/0046364; and PCT Application Publication No. WO 2011/038207, all of which are hereby incorporated by reference in their entirety.

pharmaceutical compositions.

Compounds suitable for the use described above may be formulated into pharmaceutical compositions for use in the treatment of the conditions described herein. Standard methods for the preparation of pharmaceutical formulations are used, such as those disclosed in Remington's The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins (2005), incorporated herein by reference in its entirety. Accordingly, some embodiments include pharmaceutical compositions comprising (a) a safe and therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier, diluent, excipient, or combination thereof.

The term pharmaceutically acceptable carrier or pharmaceutically acceptable excipient includes any and all solvents, diluents, emulsifiers, binders, buffers, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. or any other similar compound known to those skilled in the art to be suitable for use in the preparation of pharmaceutical formulations. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional vehicle or agent is incompatible with the active ingredient, it is intended to be used in therapeutic compositions. Additional active ingredients may also be included in these compositions. In addition, various adjuvants, such as adjuvants commonly used in the art, may be included. These and other similar compounds are described in the sources, for example, Merck Index, Merck & Company, Rahway, NJ. Considerations regarding the inclusion of various components in pharmaceutical compositions are described, for example, in Gilman et al. (eds.) (1990); Goodman and Gilman's: The Pharmacological Basis of Therapeutics, 8th ed., Pergamon Press.

Some examples of substances that can serve as pharmaceutically acceptable carriers or components thereof are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and methylcellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter; polyols such as propylene glycol, glycerin, sorbitol, mannitol and polyethylene glycol; alginic acid; emulsifiers such as Twins; wetting agents such as sodium lauryl sulfate; coloring agents; flavoring agents; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline and phosphate buffer solutions.

The choice of a pharmaceutically acceptable carrier for use in conjunction with the target compound is determined by the route of administration of said compound.

The compositions described herein are preferably presented in unit dosage form. In the context of the present description, a unit dosage form is a composition containing an amount of a compound that is suitable for administration to a subject, in a single dose in accordance with good medical practice. The receipt of a single or single dosage form, however, does not mean that the specified dosage form is administered once a day or once per course of therapy. A single dosage form may include a single daily dose or a fractional sub-dose (sub-dose), while several single dosage forms must be administered to receive a full daily dose during the day. According to the present description, a single dosage form may be administered more or less than once a day, and may be administered more than once during the course of therapy. Such dosage forms may be administered by any route consistent with their formulation, including orally, parenterally, and may be administered as an infusion over a period of time (eg, from about 30 minutes to about 2-6 hours). While single administrations are specifically contemplated, compositions administered according to the methods described herein may also be administered as a continuous infusion or via an implantable infusion pump.

The methods described herein can use any of a variety of suitable forms for various routes of administration, for example, oral, nasal, rectal, topical (including transdermal), ocular, intracerebral, intracranial, intrathecal, intraarterial, intravenous, intramuscular, or other parenteral routes of administration. One of skill in the art will appreciate that compositions for oral and nasal administration include compositions that are administered by inhalation and that are prepared using available techniques. Depending on the particular route of administration desired, a variety of pharmaceutically acceptable carriers well known in the art may be used. Pharmaceutically acceptable carriers include, for example, solid or liquid excipients, diluents, hydrotropes, surfactants, and encapsulating agents. Optional pharmaceutically active agents may be included that do not substantially interfere with the activity of the compound. The amount of carrier used in combination with the compound is sufficient to provide a practical amount of substance to be administered per unit dose of said compound. Methods and compositions for preparing dosage forms suitable for use in the methods described herein are described in the following references, all of which are incorporated herein by reference: Modern Pharmaceutics, 4th ed., chapters 9 and 10 (Banker & Rhodes, editors, 2002); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1989) and Ansel, Introduction to Pharmaceutical Dosage Forms 8th Edition (2004).

Various oral dosage forms may be used, including solid forms such as tablets, capsules, granules, and bulk powders. Tablets may be compressed, may be trituration tablets, may be enteric-coated, sugar-coated, film-coated or may be obtained by multiple compression (multiple-compressed), may contain suitable binders, lubricating agents, diluents, disintegrating agents , coloring agents, flavoring agents, flow agents, and melting agents. Liquid dosage forms for oral administration include aqueous solutions, emulsions, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules and contain suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners , melting agents, coloring agents and flavoring agents.

Pharmaceutically acceptable carriers suitable for the preparation of unit dosage forms for oral administration are well known in the art. Tablets usually contain conventional pharmaceutically compatible adjuvants such as inert diluents such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders such as starch, gelatin and sucrose; disintegrants such as starch, alginic acid and croscarmellose; lubricants such as magnesium stearate, stearic acid, microcrystalline cellulose, carboxymethyl cellulose and talc. Tablets may also contain solubilizers or emulsifiers such as poloxamers, Cremophor/Kolliphor/Lutrol®, methylcellulose, hydroxypropyl methylcellulose, or other substances known in the art. Glidants such as silica can be used to improve the flow characteristics of the powder mixture. Coloring agents such as food, drug and cosmetic (FD&C, food, drugs and cosmetic) dyes may be added to provide appearance. Sweetening and flavoring agents such as aspartame, saccharin, menthol, peppermint and fruit flavors are valuable adjuvants for chewable tablets. Capsules usually contain one or more of the solid diluents disclosed above. The choice of carrier components depends on secondary considerations such as taste, cost, and storage stability, which can easily be made by one skilled in the art.

Compositions for oral (PO) administration also include liquid solutions, emulsions, suspensions, and the like. Pharmaceutically acceptable carriers suitable for the preparation of such compositions are well known in the art. Typical carrier components for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. For suspensions, typical suspending agents include methylcellulose, sodium carboxymethylcellulose, AVICEL RC-591 (AVICEL RC-591), tragacanth, and sodium alginate; typical wetting agents include lecithin and polysorbate 80; and typical preservatives include methyl paraben and sodium benzoate. Liquid compositions for oral administration may also contain one or more of the ingredients, such as sweeteners, flavoring agents, and coloring agents, as disclosed above.

Such compositions can also be coated using conventional methods, usually pH or time dependent coatings, so that the target compound is released in the gastrointestinal tract in close proximity to the desired topical application or at different times to prolong the desired action. Such dosage forms typically contain, but are not limited to, one or more of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate, ethylcellulose, Eudragit coatings, waxes, and shellac.

The compositions described herein may optionally contain other active drug substances.

Other compositions suitable for use in achieving systemic delivery of target compounds include sublingual, buccal and nasal dosage forms. Such compositions typically contain one or more soluble excipients such as

- 37 042390 sucrose, sorbitol and mannitol, and binders such as gum arabic (acacia), microcrystalline cellulose, carboxymethylcellulose and hydroxypropylmethylcellulose. The glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.

The liquid composition, which is designed for topical ophthalmic use, is prepared in such a way that it can be administered topically to the eye. Comfort can be maximized as far as possible, although sometimes it may be necessary for reasons of preparation (eg stability of the drug) that the comfort is less than optimal. In the event that comfort cannot be maximized, the liquid may be formulated to be tolerated by the patient for topical ophthalmic application. In addition, the ophthalmologically acceptable liquid may either be packaged for single use or may contain a preservative to prevent contamination over multiple uses.

Solutions or drugs for ophthalmic use are often prepared using saline as the main carrier medium. Ophthalmic solutions can preferably be maintained at a comfortable pH with a suitable buffer system. The compositions may also contain conventional pharmaceutically acceptable preservatives, stabilizers and surfactants.

Preservatives that may be used in the pharmaceutical compositions disclosed herein include, but are not limited to, benzalkonium chloride, polyhexamethylene biguanide (PHMB), chlorobutanol, thimerosal, phenylmercuric acetate, and phenylmercuric nitrate. A suitable surfactant is, for example, Tween 80. In addition, various suitable carrier media can be used in the ophthalmic formulations disclosed herein. These carrier media include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropylmethylcellulose, poloxamers, carboxymethylcellulose, hydroxyethylcellulose, and purified water.

Tonicity controls may be added as needed or for convenience. These include, but are not limited to, salts such as sodium chloride, potassium chloride, mannitol and glycerin, or any other suitable ophthalmologically acceptable tonicity regulator.

Various buffers and pH adjusters can be used, provided that the resulting formulation is ophthalmologically acceptable. For many formulations, the pH will be between 4 and 9. Suitable buffers include acetate buffers, citrate buffers, phosphate buffers, and borate buffers. Acids or bases may be used to adjust the pH of these formulations, if necessary.

Ophthalmologically acceptable antioxidants include, but are not limited to, sodium metabisulphite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole, and butylated hydroxytoluene.

Other auxiliary components that can be included in ophthalmic preparations are chelating agents. A suitable chelating agent is disodium edetate, although other chelating agents may be used instead or in combination with it.

For topical use, including transdermal administration, creams, ointments, gels, solutions or suspensions, etc. containing the compound disclosed in the present description are used. Topical formulations may generally consist of a pharmaceutical carrier, a co-solvent, an emulsifier, a penetration enhancer, a preservative system, and an emollient.

For intravenous administration, the compounds and compositions described herein may be dissolved or dispersed in a pharmaceutically acceptable diluent such as saline or dextrose solution. Suitable excipients may be included to achieve the desired pH, including but not limited to NaOH, sodium carbonate, sodium acetate, HCl, and citric acid. In various embodiments, the pH of the finished composition is from 2 to 8, or preferably from 4 to 7. Antioxidant excipients may include sodium bisulfite, sodium acetone bisulfite, sodium sulfoxylate formaldehyde, thiourea, and EDTA. Other non-limiting examples of suitable adjuvants that may be found in the intravenous formulation may include sodium or potassium phosphate, citric acid, tartaric acid, gelatin, and carbohydrates such as dextrose, mannitol, and dextran. Other acceptable excipients are described in Powell, et al., Compendium of Excipients for Parenteral Formulations, PDA J. Pharm. sci. and Tech., 1998, 52, 238-311; and Nema et al., Excipients and Their Role in Approved Injectable Products: Current Usage and Future Directions, PDA J. Pharm. sci. Tech., 2011, 65, 287332, both of which are incorporated herein by reference in their entirety. Antimicrobial agents may also be included to provide a bacteriostatic or fungistatic solution, including but not limited to phenylmercury nitrate, thimerosal, benzethonium chloride, benzalkonium chloride, phenol, cresol, and chlorobutanol.

Compositions for intravenous administration may be provided to caregivers

- 38 042390 for patients, in the form of one or more solids, which are reconstituted with a suitable diluent such as sterile water, saline or dextrose in water shortly before administration. In other embodiments, said compositions are provided in a solution ready for parenteral administration. In other embodiments, said compositions are provided in a solution that is further diluted prior to administration. In embodiments that involve administering a combination of a compound described herein and another agent, said combination may be provided to caregivers as a mixture, or caregivers may mix said two agents prior to administration, or said two agent can be administered separately.

The actual unit dose of the active compounds described herein depends on the particular compound and the condition being treated. In some embodiments, the dosage may be from about 0.01 mg/kg to about 120 mg/kg of body weight or more, from about 0.05 mg/kg or less to about 70 mg/kg, from about 0.1 mg /kg to about 50 mg/kg body weight, from about 1.0 mg/kg to about 10 mg/kg body weight, from about 5.0 mg/kg to about 10 mg/kg body weight, or from about 10.0 mg/kg to about 20.0 mg/kg body weight. In some embodiments, the indicated dose may be less than 100, 90, 80, 70, 60, 50, 40, 30, 25, 20, 10, 7.5, 6, 5, 4, 3, 2.5, 1, 0 .5, 0.1, 0.05 or 0.005 mg/kg body weight. In some embodiments, the actual unit dose is 0.05, 0.07, 0.1, 0.3, 1.0, 3.0, 5.0, 10.0, or 25.0 mg/kg body weight. Thus, when administered to a 70 kg individual, the dosage range would be from about 0.1 to 70 mg, from about 1 mg to about 50 mg, from about 0.5 mg to about 10 mg, from about 1 mg to about 10 mg. , from about 2.5 mg to about 30 mg, from about 35 mg or less to about 700 mg or more, from about 7 mg to about 600 mg, from about 10 mg to about 500 mg, or from about 20 mg to about 300 mg, or from about 200 mg to about 2000 mg. In some embodiments, the actual unit dose is 5 mg. In some embodiments, the actual unit dose is 10 mg. In some embodiments, the actual unit dose is 25 mg. In some embodiments, the actual unit dose is 250 mg or less. In some embodiments, the actual unit dose is 100 mg or less. In some embodiments, the actual unit dose is 70 mg or less.

These compounds may also be included in formulations for delivery outside the systemic circulation. Such formulations may include enteric-coated capsules, tablets, soft gels, spray-dried powders, polymer matrices, hydrogels, enteric-coated solids, crystalline solids, amorphous solids, vitreous solids, coated micronized particles, sprayable liquids. liquids, aerosols or microcapsules.

Methods of administration.

The compositions described above may be administered using any suitable route of administration, for example by injection, for example subcutaneously, intramuscularly, intraperitoneally, intravenously or intraarterially; topically, for example, with a cream, lotion or patch; orally, for example, by means of a pill, a diluted liquid, an oral suspension, a buccal film, or a mouthwash; nasally, for example, using a nasal aerosol, powder or spray; or ocularly, for example, using eye drops. In some embodiments, said composition may be administered once, twice, three times, or four times per day. In other embodiments, said composition may be administered once, twice, or three times per week. In other embodiments, said composition is administered every other day, once every three days, or once every four days. In other embodiments, said composition is administered once every two weeks, once every three weeks, or once every four weeks. In other embodiments, said composition is administered once a month or twice a month.

In some embodiments, an initial loading dose is administered that is higher than subsequent doses (maintenance doses). The dosage form or route of administration for the maintenance dose may be different from the dosage form or route of administration used for the loading dose. In any of the embodiments disclosed herein, administration of a maintenance dose may include administration of a single dosage form according to any dosage regimen described herein, including, but not limited to, administration monthly or several times a month, once every two weeks or several times in two weeks, weekly or several times a week, daily or several times a day. In the present description, it is assumed that drug holidays (dosing holidays) can be included in the period of administration of the maintenance dose. Such drug holidays may occur immediately after the administration of the loading dose or at any time during the period of administration of the maintenance dose. In some embodiments, the loading dose is 300 mg or less, 250 mg or less, 200 mg or less, 150 mg or less, or 100 mg or less. In some embodiments, the maintenance dose is 300 mg or less, 200 mg or less, 100 mg or less, 50 mg or less, 25 mg or less, 10 mg or less, 5 mg or less, or 1 mg or less.

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Methods of treatment.

Some embodiments of the methods and compositions described herein relate to a method of reducing or preventing the deposition of extracellular matrix proteins, comprising administering an effective amount of a compound described herein to a subject in need thereof. In some embodiments, said deposition of extracellular matrix proteins may include abnormal or excessive deposition of said proteins. In some embodiments, said extracellular matrix proteins may include one or more of collagen, keratin, elastin, or fibrin. In some embodiments, said extracellular matrix proteins may include collagen. In some embodiments, these extracellular matrix proteins may include type I collagen. In some embodiments, these extracellular matrix proteins may include type Ia collagen. In some embodiments, said extracellular matrix proteins may include type III collagen. Some embodiments of the compositions and methods described herein relate to a method of treating fibrosis or its symptoms or consequences, comprising administering an effective amount of a compound described herein to a subject in need thereof.

In some embodiments, the compounds and compositions containing the compounds described herein may be used to treat a variety of conditions resulting from fibrosis or inflammation and specifically including conditions associated with abnormal collagen deposition. Illustrative conditions include glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), non-alcoholic steatohepatitis (NASH), cirrhosis, hepatitis, scleroderma, alcoholic fatty liver disease, atherosclerosis, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, liver and gallbladder cirrhosis, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, chronic kidney disease, type II diabetes, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, injection fibrosis, post-surgical complications, fibrotic chronic allograft vasculopathy and/or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, post-vasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis, Dupuytren's disease, dermatomyositis-polymyositis, mixed connective tissue disease, fibrous proliferative lesions of the oral cavity, fibrosing bowel strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, radiation-induced fibrosis, fibrosis-induced fibrosis breast cyst rupture, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis or their symptoms or consequences, or other diseases or conditions leading to excessive deposition of extracellular matrix components such as collagen.

In some embodiments, the methods of the present disclosure include methods for treating, improving, or preventing a fibrotic condition. In some embodiments, said fibrotic condition may be secondary to another condition. In some embodiments, said fibrotic condition or primary condition may also include chronic inflammation of an organ, tissue, spatial region, or fluid-connected area of the subject's body. In some embodiments, said inflammation may include activation of one or more TGF-beta dependent signaling pathways. In some embodiments, these TGF-B dependent signaling pathways may contain one or more T3 or T4 responsive elements. In some embodiments, said fibrotic condition may include abnormal or excessive deposition of one or more of collagen, keratin, or elastin. In some embodiments, said fibrotic condition may include abnormal or excessive collagen deposition. In some embodiments, said fibrotic condition may include abnormal or excessive deposition of type I collagen. In some embodiments, said fibrotic condition may include abnormal or excessive deposition of type Ia collagen. In some embodiments, said fibrotic condition may include abnormal or excessive deposition of type III collagen. In some embodiments, said fibrotic condition may include one or more of glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), non-alcoholic steatohepatitis (NASH), cirrhosis , hepatitis, scleroderma, alcoholic fat

- 40 042390 liver disease, atherosclerosis, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, cirrhosis of the liver and gallbladder, spleen fibrosis, scleroderma, pulmonary fibrosis, idiopathic pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia , pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, x chronic kidney disease, type II diabetes, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, fibrosis after injection, complications after surgery, fibrous chronic allograft vasculopathy and/or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, postvasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis, Dupuytren's disease, polymyositis dermatomyositis, mixed connective tissue disease, fibrous proliferative lesions of the oral cavity, fibrosing bowel strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, fibrosis, induced by radiation exposure, fibrosis caused by rupture of a breast cyst, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis. In some embodiments, said fibrotic condition may include one or more of GSD III, GSD IX, non-alcoholic steatohepatitis, liver and/or pancreatic cirrhosis, scleroderma, idiopathic pulmonary fibrosis, psoriasis, alcoholic fatty liver disease, Dupuytren's disease, and/or any of them. combinations.

According to the methods and compositions of the present disclosure, thyroid receptor agonists, such as the agonists disclosed herein and particularly comprising Compounds 1-4, may be administered to a subject to treat, ameliorate, prevent, or cure a fibrotic condition or a condition in which fibrosis is a symptom or consequence. According to the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may also include chronic inflammation. According to the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may also include activation of one or more TGF-β-dependent signaling pathways. According to the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may also include activation and/or repression of one or more thyroid receptor beta (TRe) dependent signaling pathways. According to the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may also involve involvement of signaling pathways sensitive to triiodothyronine (T3), thyroxine (T4), any combination thereof, or mimetics thereof. According to the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may also involve involvement of receptors sensitive to T3, T4, any combination thereof, or mimetics thereof. In some embodiments of the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may involve TRe. In some embodiments of the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may include one or more conditions that are prevented, ameliorated, or cured by administration of one or more TRe agonists. In some embodiments of the methods and compositions disclosed herein, said fibrotic condition or condition resulting in fibrosis may include one or more conditions that are prevented, ameliorated, or cured by administration of one or more of Compounds 1-4. In some embodiments, said one or more TRe agonists or said one or more of Compounds 1-4 may be co-administered with one or more active drug compounds and/or one or more excipients. In some embodiments, said one or more TRe agonists or said one or more of Compounds 1-4 may be administered before, during, or after surgery, phototherapy, or ultrasound therapy. In some embodiments, said one or more TRe agonists or said one or more of Compounds 1-4 may be co-administered with one or more of Pirfenidone, nintedanib, and/or a fibroblast growth factor receptor antagonist, and/or a collagenase such as collagenase Clostridium histolyticum.

In some embodiments of the compositions and methods described herein, compositions and methods are provided for treating, improving, preventing, or curing collagen deposition. In some embodiments, said collagen deposition includes and

- 41 042390 abnormal or excessive collagen deposition. In some embodiments, said collagen deposition may include abnormal or excessive type I collagen deposition. In some embodiments, said collagen deposition may include abnormal or excessive type Ia collagen deposition. In some embodiments, said collagen deposition may include abnormal or excessive type III collagen deposition. According to the methods and compositions disclosed in the present description, the specified deposition of collagen may also include the participation of receptors sensitive to T3, T4, any combination thereof or their mimetics. In some embodiments of the methods and compositions disclosed herein, said collagen deposition may involve TRe. In some embodiments of the methods and compositions disclosed herein, said collagen deposition can be prevented, attenuated, or treated by administering one or more TRe agonists. In some embodiments of the methods and compositions disclosed herein, said collagen deposition can be prevented, attenuated, or treated by administering one or more of Compounds 1-4. In some embodiments, said one or more TRe agonists or said one or more of Compounds 1-4 may be co-administered with one or more active drug compounds and/or one or more excipients. In some embodiments, said one or more TRe agonists or said one or more of Compounds 1-4 may be administered before, during, or after surgery, phototherapy, or ultrasound therapy. In some embodiments, said one or more TRe agonists or said one or more of Compounds 1-4 may be co-administered with one or more of Pirfenidone, nintedanib, and/or a fibroblast growth factor receptor antagonist, and/or a collagenase such as collagenase Clostridium histolyticum.

In some embodiments, administration of Compounds 1-4, Compound 2, or any of the compounds or compositions disclosed herein results in decreased expression of the Cola1, Col3a1, aSMA, and/or Galectinl genes, or any combination or product thereof, in a subject administered said connection or composition. In some embodiments, administration of Compounds 1-4, Compound 2, or any of the compounds or compositions disclosed herein results in a reduction in the degree of fibrosis as observed by histology, histochemistry, immunohistochemistry, or the like, and/or a reduction in the amount, accumulation, or distribution of type I collagen and/or hydroxyproline, or any combination thereof, in a subject to whom said compound or composition is administered. In some embodiments, administration of Compounds 1-4, Compound 2, or any of the compounds or compositions disclosed herein results in a decrease in total serum lipids, total serum cholesterol, total serum triglycerides, total liver lipids, total cholesterol liver, total liver triglycerides, or any combination thereof.

The methods described herein are further illustrated by the following examples.

Example 1

DIO-NASH mice were acclimatized for 3 weeks, pre-liver biopsy specimens were collected prior to acclimatization. Mice were randomly assigned to one of five dosing groups, 12 mice per group. Doses prescribed: Compound 2 (low dose) 3 mg/kg; compound 2 (high dose) - 10 mg/kg; compound 1 - 10 mg/kg; and elafibranor (30 mg/kg). One group received sham treatment with vehicle control only. Dosage forms were administered orally once a day. After 8 weeks, the animals were sacrificed and liver samples were taken. Liver samples were analyzed for total liver hydroxyproline and subjected to immunohistochemical analysis to determine the stage of fibrosis, as well as the degree of staining Cola1 (collagen type Ia). As shown in FIG. 1, Compound 2-treated animals exhibited lower levels of total hydroxyproline in the liver than control or sham treated animals. Because hydroxyproline is a significant component of collagen, and collagen is the most significant source of hydroxyproline in animal tissue, hydroxyproline levels provide a reliable indication of the presence of collagen in a sample.

Samples of the final liver biopsy were also subjected to histochemical staining and immunohistochemical staining. Representative images of the liver stained with Picro-Sirius Red (to visualize collagen I and III deposition, red spots) at the end of treatment after 8 weeks of vehicle, low dose Compound 2, high dose Compound 2, Compound 1 or Elafibranor are shown in FIG. 2. As shown in FIG. 5, total liver collagen 1 and 3 (mg/liver) was determined by morphometry after Picro-Sirius Red staining. Liver sections from Compound 1 treated animals showed lower PSR staining than from sham treated animals; Liver sections from Compound 2 treated animals showed lower PSR staining than control or sham treated animals. Representative images of a liver stained against type I collagen (col1a1) (Southern Biotech, Cat. 131001) at the end of the treatment period after 8 weeks of vehicle, low dose Compound 2, high dose Compound 2, Compound 1, or elafib-42 042390 ranor are shown in fig. 3. The degree of ColAl content was calculated as the total ColAl staining of the liver in the samples of the final liver biopsy. As shown in FIG. 4, in animals treated with compound

2, the total content of ColA1 in the liver was lower than in animals that were administered control or sham treatment.

Fibrosis scores were also calculated based on observation of end liver biopsy specimens. In addition, a higher proportion of animals treated with Compound l or Compound 2 showed a reduction in fibrosis score after treatment compared to animals treated with sham treatment. Animals treated with Compound 2 did not show an increase in fibrosis score after treatment.

Example 2

In healthy male Dunkin-Hartley guinea pigs, pulmonary fibrosis was induced by intratracheal administration of bleomycin. Control subjects were subjects who received intratracheal saline administration. After the establishment of pulmonary fibrosis in animals that received bleomycin, experimental preparations containing any of the compounds 1-4 or any other compound disclosed herein were administered to each subject in accordance with its composition, daily or as needed for 6-10 weeks. Unilateral lung biopsies were taken before the first administration of the experimental drugs and again after sacrifice after the last administration of the experimental drugs. Biopsy samples were analyzed as described in example I, with the addition of immunohistochemical staining for type III collagen. The lungs of animals treated with the compounds disclosed herein, especially the lungs of animals treated with compound 2, showed reduced levels of hydroxyproline, reduced collagen III staining, and a reduced fibrosis score relative to levels shown prior to administration of the experimental drugs. Sham treated animals showed little or no reduction in fibrosis, hydroxyproline, or collagen III.

Example 2a.

Fibrosis of the palmar fascia was induced in athymic mice by injecting fibroblasts from the fibrous cords of Dupuytren's disease patients as described in Stish, L. et al, BMC Musculoskelet. Disord., 16:138-148 (2015), which is incorporated herein by reference with respect to its description of the establishment of an animal system model for the study of volar fascia fibrosis. After fibrosis of the palmar fascia was established in fibroblast-treated animals, each subject was administered experimental formulations containing any of Compounds 1-4 or any other compound disclosed herein, according to its formulation, daily or as needed for 6-10 weeks. Unilateral forepaw biopsies were taken before the first administration of the experimental drugs and again after sacrifice after the last administration of the experimental drugs. Biopsy samples were analyzed as described in Example 1 with the addition of immunohistochemical staining for type III collagen. In the palmar fascia of animals treated with the compounds described herein, especially those treated with Compound 2, there was a decrease in hydroxyproline levels, a decrease in collagen III staining, and a decrease in fibrosis score relative to levels shown prior to type administration. Sham treated animals showed little or no reduction in fibrosis, hydroxyproline, or collagen III.

Example 3

In Sprague Dawley rats, subcutaneous injection of capsaicin induced hypertrophic skin lesions as described in Wallengren, J. et al., Skin Pharm. Appl. Skin Physiol., 15(3):154-165 (2002), which is incorporated herein by reference with respect to describing the induction of hypertrophic skin lesions in rats; or in C57BL or other appropriate mouse strains by subcutaneous administration of CCl ₄ and/or bleomycin as described in Alonso-Merino et al., Proc. Nat. Acad. Sci., 113(24):E3451-60 (2016), which is incorporated herein to disclose information on the induction of fibrotic skin lesions in mice. After the establishment of hypertrophic skin lesions in animals treated with capsaicin, CCl ₄ and/or bleomycin, experimental preparations containing any of the compounds 1-4 or any other compound disclosed herein were administered to each animal subject in accordance with its composition daily or as needed for 6-10 weeks. Skin biopsies from the injection site were taken before the first administration of the experimental drugs and again after sacrifice after the last administration of the experimental drugs. Biopsy samples were analyzed as described in Example 1 with the addition of immunohistochemical staining for type III collagen. Skin samples at the injection site from animals that received the compounds disclosed herein, especially from animals that received compound 2, showed reduced levels of hydroxyproline, reduced collagen III staining, and a reduced fibrosis score relative to levels shown before administration of the experimental drugs. Sham treated animals showed little or no reduction in fibrosis, hydroxyproline, or collagen III.

- 43 042390

Example 4

Glucose-6-phosphatase-a deficient mice exhibiting GSD-3-like hepatic symptoms including hypercholesterolemia and hyperlipidemia (Agl-/-, see e.g. Liu, K. M. et al., Mol. Genet. Metabol. , 111(4):467-76 (2014)) received experimental preparations containing any of the compounds 1-4 or any other compound disclosed herein, and each subject was administered according to its composition, daily or as needed in within 6-10 weeks. Liver biopsies were taken before the first administration of experimental drugs and again after sacrifice after the last administration of experimental drugs. Biopsy samples were analyzed as described in Example I. Liver samples from animals that received the compounds described herein, especially those animals that received compound 2, showed reduced levels of hydroxyproline, reduced collagen I staining, and a reduced fibrosis score relative to levels shown before administration of experimental drugs. Sham treated animals showed little or no reduction in fibrosis, hydroxyproline, or collagen I.

Example 5

Phosphorylase kinase-deficient mice exhibiting GSD-8/9-like hepatic symptoms including hypercholesterolemia and hyperlipidemia (PhKc-/-, see e.g. Varsanyi, M. et al., Biochem. Genet., 18(3 -4):247-61 (1980)), received experimental preparations containing any of the compounds 1-4 or any other compound disclosed herein, and each subject was administered according to its composition, daily or as needed for 6-10 weeks. Liver biopsies were taken before the first administration of experimental drugs and again after sacrifice after the last administration of experimental drugs. Biopsy samples were analyzed as described in Example 1. Liver samples from animals that received the compounds described herein, especially those animals that received compound 2, showed reduced levels of hydroxyproline, reduced collagen I staining, and a reduced fibrosis score relative to levels shown before administration of experimental drugs. In animals treated with sham treatment, there was a slight decrease in fibrosis, hydroxyproline or collagen I levels.

Example 6

Compound 2 was evaluated in a diet-induced NASH mouse model (see, e.g., Hansen, H. et al., Drug Discovery Today, 22(17):1701-1718 (2017), which is incorporated herein by reference with respect to the disclosure information on diet-induced, genetic, chemical and other models of NASH in mice). The model line of diet-induced NASH mice does not rely on chemical/toxin exposure to generate steatohepatitis/fibrosis. Animals were biopsied prior to study and only animals with NASH and fibrosis were selected for study. Selected animals were acclimated and randomized, with experimental groups of 11-12 animals per group receiving oral dosing of Compound 2 according to the following schedules: daily dosing for 8 weeks; or daily dosing for weeks 0-1 followed by weekly dosing for weeks 2-8. At week 8, the animals were sacrificed and tissues analyzed. Plasma enzymes (P-ALT (alanine aminotransferase) and P-AST (aspartate aminotransferase)), plasma total triglycerides, and plasma total cholesterol were measured and a final autopsy of each liver was performed, analyzing total liver biochemistry including total liver triglycerides, total liver cholesterol, as well as histological assessment of NAFLD activity score (from the English. Nonalcoholic fatty liver disease; non-alcoholic fatty liver disease) (before and after treatment), fibrosis stage (also before and after treatment), steatosis, Col1a1 level and galactin-3 level. Tissue samples were stored for characterization using RNA sequencing; RNA sequencing was used to determine expression levels of genes showing differential expression in Compound 2 treated and vehicle treated animals and/or genes known to be involved in fibrosis. Significant reductions in hepatic triglycerides and cholesterol levels were observed in treated groups compared to control groups. As shown in FIG. 6, total liver lipids were reduced by about 80% with a similar reduction in plasma lipids and a significant improvement in NAS scores. No significant toxicity was observed.

As shown in FIG. 7, significant reductions in fibrosis, collagen type I deposition, and hydroxyproline (by 50.2%, 60.2%, and 46.3%, respectively) were also observed relative to pre-treatment samples. After treatment, expression of the profibrotic genes Col1a1, Col3a1, aSMA and Galectin 1 was reduced by 36.3%, 27.1%, 37% and 64.7%, respectively (FIG. 8), confirming the histological findings. Thus, 8 weeks of Compound 2 administration resulted in marked improvements in histological, biochemical, and genetic markers associated with steatosis, fibrosis, and non-alcoholic steatohepatitis in a diet-induced NASH model.

With regard to the use in this document of any terms in the plural and/or

- 44 042390 singular, those skilled in the art may translate them from the plural to the singular and/or from the singular to the plural, depending on the context and/or application. The various singular/plural permutations may be explicitly stated here for the sake of clarity.

Those skilled in the art will appreciate that, as a general rule, terms used herein, and especially in the appended claims (e.g., in the text of the appended claims), are generally intended to be open terms (e.g., the term including should be interpreted as including , but not limited to, the term having should be interpreted as having at least, the term includes should be interpreted as including, but not limited to, etc.). In addition, those skilled in the art will appreciate that if the wording of a claim is intended to indicate a specific number, that intent will be explicitly stated in the claims, and in the absence of such an indication, there is no such intent. For example, to facilitate understanding, in the following appended claims, introductory phrases of at least one and one or more may be used to introduce features of the claims. However, the use of such phrases should not be construed as implying that the introduction of a feature in an unspecified form limits any particular claim containing such introduction of a feature to embodiments containing only one such feature, even when that claim includes introductory the phrases one or more or at least one and the indefinite singular (eg, the indefinite singular should be interpreted as at least one or one or more); the same applies to the use of certain forms used to state the features of the claims. In addition, even if a claim specifies an amount explicitly, those skilled in the art will appreciate that such an indication is to be interpreted as meaning at least the number indicated (e.g., a mere statement of two indications without other modifiers means at least two repetitions, or two or more repetitions). Moreover, in cases where an indication similar to at least one of A, B, C, etc. is used, in general, such a construction is presented in the sense that a person skilled in the art should understand this indication (for example , a system having at least one of A, B, and C would include, but not be limited to, systems having A only, B only, C only, A and B together, A and C together, B and C together, and/ or A, B and C together, etc.). Where an indication similar to at least one of A, B, or C, etc. is used, such a construction is presented in the sense that a person skilled in the art should understand this indication (for example, a system having at least one of A, B, or C may include, but is not limited to, systems that have A only, B only, C only, A and B together, A and C together, B and C together, and/or A, B and C together, etc.). Furthermore, those skilled in the art will appreciate that virtually any separating word and/or phrase representing two or more alternative terms, whether in the description, claims, or drawings, should be understood to encompass the possibility of including one of the terms, any from terms or two terms at once. For example, the phrase A or B will be understood to include the possibilities of A or B or A and B.

Moreover, if features or aspects of a disclosure are described in terms of Markush groups, those skilled in the art will appreciate that the disclosure is thus described in terms of any individual member or subgroup of members of a Markush group.

As will be appreciated by one of skill in the art, for any and all purposes, such as providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of their subranges. Any of the listed ranges can be easily recognized as being sufficiently descriptive and allowing one and the same range to be broken down into at least equal halves, thirds, quarters, fifths, tenths, and so on. As a non-limiting example, each range discussed herein can be easily broken down into lower third, middle third, and upper third, and so on. As will be appreciated by one skilled in the art, all expressions such as before, at least more, less, etc. include the number listed and refer to ranges that may subsequently be sub-ranged as discussed above. Finally, as one of skill in the art will appreciate, the range includes every single element. So, for example, a group consisting of 1-3 elements refers to groups having 1, 2 or 3 elements. Similarly, a group having 1-5 elements refers to groups having 1, 2, 3, 4, or 5 elements, and so on.

Although various aspects and implementations are disclosed in the present description, other aspects and implementations will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are intended to be illustrative and not intended to be limiting, the true scope and spirit being set forth in the following claims.

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Claims (6)

CLAIM 1. A method for the therapeutic treatment of fibrosis resulting in a reduction in the amount of extracellular matrix proteins present in one or more tissues of a subject, comprising administering to a subject in need thereof one or more compounds having a structure selected from the group consisting of or their pharmaceutically acceptable salts. 2. The method of claim 1, wherein said subject has one or more fibrotic conditions selected from glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX) , hepatitis, scleroderma, atherosclerosis, asthma, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, gallbladder cirrhosis, spleen fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, diffuse parenchymal lung disease, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial pneumonitis, desquamative interstitial pneumonia, respiratory bronchiolitis, interstitial lung disease, chronic interstitial lung disease, acute interstitial pneumonitis, hypersensitivity pneumonitis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, lymphocytic interstitial pneumonia, pneumoconiosis, silicosis, emphysema, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, chronic kidney disease, type II diabetes, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, fibrosis after injections, complications after surgery, fibrous chronic allograft vasculopathy and/or chronic organ transplant rejection, fibrosis associated with ischemic reperfusion injury, postvasectomy pain syndrome, fibrosis associated with rheumatoid arthritis, arthrofibrosis, Dupuytren's disease, dermatomyositis-polymyositis, mixed connective tissue disease tissue, fibrous proliferative lesions of the oral cavity, fibrosing bowel strictures, Crohn's disease, glial scarring, leptomeningeal fibrosis, meningitis, systemic lupus erythematosus, exposure-induced fibrosis radiation, fibrosis caused by rupture of a breast cyst, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, or any combination thereof. 3. The method of claim 2 wherein said condition is primary fibrosis. 4. The method according to any one of claims 2 and 3, wherein fibrosis is a symptom of said condition. 5. The method of claim 1, wherein said subject has one or more of scleroderma, atherosclerosis, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, bone marrow fibrosis, liver fibrosis, spleen fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial lung disease, chronic interstitial lung disease, pneumoconiosis, silicosis, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, atrial fibrosis, endomyocardial fibrosis, renal fibrosis, macular degeneration, keloid lesions, hypertrophic scar, nephrogenic systemic fibrosis, fibrosis after injection, fibrotic complications after surgery, fibrous chronic allograft vasculopathy, fibrosis associated with ischemic reperfusion injury, arthrofibrosis, Dupuytren's disease, fibrous proliferative lesions of the oral cavity cavities, fibrosing bowel strictures, glial scarring, leptomeningeal fibrosis, radiation-induced fibrosis, breast cyst rupture fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, or any combination thereof. 6. The method of claim 1, wherein said subject has one or more of glycogen storage disease type III (GSD III), glycogen storage disease type VI (GSD VI), glycogen storage disease type IX (GSD IX), hepatitis, scleroderma , atherosclerosis, asthma, cirrhosis of the gallbladder, diffuse parenchymal lung disease, interstitial pneumonitis, desquamative interstitial -