Treatment of gastrointestinal diseases

Abstract

Disclosed herein is the use of thiazolidinediones for the treatment of gastrointestinal disorders, including gastroparesis.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 332,778, filed April 20, 2022, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates to therapeutic agents and methods for the treatment of gastrointestinal disorders such as gastroparesis. [Background technology]

[0003] A digestive disorder is a disorder of the digestive tract, also known as the gastrointestinal (GI) tract. A GI disorder or disease is any illness related to the GI system, which includes the entire GI tract from the mouth to the anus, including the esophagus, stomach, and intestines. Diagnoses may include acute illnesses, or illnesses that are short-term. GI disorders may also include more chronic diagnoses, which may require long-term, specialized treatment.

[0004] Conditions range from mild to severe. Some common problems include heartburn, cancer, irritable bowel syndrome (IBS), and lactose intolerance. Other digestive diseases include gallstones, cholecystitis, cholangitis, rectal diseases (e.g., anal fissures, hemorrhoids, proctitis, and rectal prolapse), esophageal diseases (e.g., strictures and achalasia and esophagitis), gastric diseases (e.g., gastritis, gastric ulcers and cancer usually caused by Helicobacter pylori infection), liver diseases (e.g., hepatitis B or C, cirrhosis, liver failure, and autoimmune and alcoholic hepatitis), pancreatitis and pancreatic pseudocysts, intestinal diseases (e.g., polyps and cancer, infections, celiac disease, Crohn's disease, ulcerative colitis, diverticulitis, malabsorption, short bowel syndrome, and intestinal ischemia), gastroesophageal reflux disease (GERD), peptic ulcer disease, and hiatal hernia.

[0005] Among all digestive diseases, gastroparesis is a disorder characterized by delayed gastric emptying (DGE) in the absence of mechanical obstruction. Symptoms are chronic with episodic exacerbations. The most common idiopathic form of the disorder affects mainly young adult women. Gastroparesis is also frequently associated with diabetes (diabetic gastroparesis), likely due to impaired neural control of gastric motility. In addition, acute hyperglycemia can slow gastric emptying and reduce the effectiveness of prokinetic drugs.

[0006] At the molecular level, gastroparesis may be caused by the loss of neuronal nitric oxide expression, as cells in the GI tract secrete nitric oxide. This important signaling molecule has a variety of responsibilities in the GI tract and muscles throughout the body. When nitric oxide levels are low, smooth muscles and other organs may not be able to function properly. Another important component of the stomach is the interstitial cells of Cajal (ICC), which act as pacemakers by transducing signals from motor neurons to generate electrical potential rhythms in smooth muscle cells. Low nitric oxide levels also correlate with the loss of ICC cells, which may ultimately result in the loss of smooth muscle function in the stomach and other areas of the GI tract. The pathogenesis of symptoms in diabetic gastroparesis includes (1) loss of nitric oxide synthase (NOS)-containing gastric neurons, which causes defective accommodative reflexes and leads to early satiety and postprandial bloating; (2) impaired electromechanical activity in the myenteric plexus, which causes delayed gastric emptying and results in nausea and vomiting; (3) sensory neuropathy in the stomach wall, which may cause epigastric pain; and (4) abnormal pacemaker activity (tachybradyalthemia), which may generate noxious signals that are transmitted to the CNS, causing nausea and vomiting.

[0007] Macrophages also play a role in the development and progression of gastroparesis. In particular, macrophages infiltrate the smooth muscle layer of the stomach in people with gastroparesis, releasing proinflammatory molecules that cause abdominal muscle damage and dysfunction. In diabetic gastroparesis, high blood glucose levels activate macrophages, leading to inflammation and damage to the nerves and muscles of the stomach. There are two different subtypes of macrophages with various functions in the immune system: M1 macrophages and M2 macrophages. M1 macrophages are involved in the inflammatory response and are activated in response to proinflammatory signals. They are important in fighting infections. M1 macrophages produce inflammatory cytokines and reactive oxygen species, killing invading pathogens and promoting the recruitment of other immune cells at the site of infection. On the other hand, M2 macrophages are involved in tissue repair and immune regulation and are activated in response to anti-inflammatory signals. They are important in resolving inflammation and promoting tissue remodeling and healing. M2 macrophages produce anti-inflammatory cytokines and growth factors that help repair damaged tissues and promote angiogenesis. Overall, M1 and M2 macrophages represent two distinct functional states of macrophages, allowing them to respond to different signals and play different roles in the immune system. The balance between M1 and M2 macrophages is important in maintaining immune homeostasis and promoting appropriate immune responses. Drugs that can inhibit M1 macrophages, promote M2 macrophages, and / or allow repolarization from M1 to M2 macrophages may be able to treat gastrointestinal diseases, including gastroparesis.

[0008] Because signs and symptoms of gastroparesis overlap with other GI conditions, gastroparesis may occasionally be misdiagnosed as ileus, functional dyspepsia, irritable bowel syndrome, or peptic ulcer disease. In patients with signs and symptoms suggestive of gastroparesis, findings of DGE without obstruction or alternative diagnoses provide important support for the diagnosis of gastroparesis and can be evaluated using either gastric emptying scintigraphy, gastric emptying breath testing, or the SmartPill™ Motility Testing System.

[0009] Approved drugs for treating gastroparesis can be divided into two different groups. The first group of drugs, including diphenhydramine and ondansetron, and prochlorperazine, only provide temporary relief for some symptoms, such as nausea and vomiting. The second group of drugs is for stimulating the abdominal muscles and includes metoclopramide (e.g., Reglan®) and erythromycin. However, both metoclopramide and erythromycin carry the risk of serious side effects. Clearly, there is an urgent medical need for the development of safe and effective therapies to treat patients with gastroparesis.

[0010] Thiazolidinediones represent a novel therapeutic strategy in diseases involving digestive disorders such as gastroparesis that can slow, halt or reverse the course of the underlying disease. Summary of the Invention

[0011] Disclosed herein is a method for treating GI disease, comprising administering to a subject in need thereof an effective amount of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone.In some embodiments, the thiazolidinedione is lobeglitazone.In some embodiments, the GI disease is gastroparesis.In some embodiments, the gastroparesis is idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis, or pharmaceutical-induced gastroparesis.

[0012] Lobeglitazone is a thiazolidinedione with the chemical name 5-(4-(2-((6-(4-methoxyphenoxy)pyrimidin-4-yl)(methyl)amino)ethoxy)benzyl)thiazolidine-2,4-dione. Lobeglitazone has been approved as an antidiabetic drug under the trade name Duvie®. As both a PPARα and PPARγ agonist, it acts as an insulin sensitizer by binding to PPAR receptors in fat cells and making the cells more responsive to insulin. Lobeglitazone has shown much greater potency in PPARγ activation than pioglitazone. Lobeglitazone is represented by the following chemical structure:

[0013] [ka]

[0014] Gastroparesis is a disorder characterized by delayed gastric emptying (DGE) in the absence of mechanical obstruction. Symptoms are chronic, with episodic exacerbations. The most common idiopathic form of the disorder primarily affects young adult women. Gastroparesis is also frequently associated with diabetes (diabetic gastroparesis), likely due to impaired neural control of gastric motility. Macrophages play a role in the development and progression of gastroparesis.

[0015] In some embodiments, the GI disorder affects the digestive tract or tissues selected from the mouth, pharynx (throat), esophagus, stomach, small intestine, large intestine, rectum, anus, salivary glands, liver, gallbladder, and pancreas.

[0016] In some embodiments, the method comprises treating a cell of the GI system selected from an absorptive cell (intestinal epithelial cell), a goblet cell, a pancreatic islet cell, a gastrointestinal endocrine cell, a hepatocyte, a Paneth cell, a fenestrated hepatic endothelial cell, a Kupffer cell, a serous cell, a gastric chief cell, a mucous cell, a smooth muscle cell, a gastric parietal cell, a myoepithelial cell, a stem cell, a gastric surface mucous cell, a pancreatic acinar cell, a taste bud, a type of interstitial cell of Cajal (ICC), and a neuronal cell.

[0017] In some embodiments, the cells are animal cells. In some embodiments, the cells are human cells. In some embodiments, the cells are treated in vitro. In some embodiments, the cells are treated ex vivo. In some embodiments, the cells are treated in vivo.

[0018] In some embodiments, the cells are used to treat conditions such as abdominal adhesions, acid reflux in adults (gastroesophageal reflux disease or GERD), acid reflux in children (GERD), anatomical problems of the lower GI tract, appendicitis, Barrett's esophagus, bowel control problems (fecal incontinence), celiac disease, colon polyps, constipation, Crohn's disease, cyclic vomiting, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, pseudomyelitis, bronch ... The disease or disorder may be in or originate from a subject having or at risk for a GI disease or disorder selected from any one or more of: chronic ileus, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcer (gastric ulcer), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, Zollinger-Ellison syndrome, or symptoms associated therewith. In some embodiments, the disease or disorder is gastroparesis. In some embodiments, the gastroparesis is idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis, or medicinal drug-induced gastroparesis.

[0019] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone to alleviate one or more signs and / or symptoms of gastroparesis. In some embodiments, the thiazolidinedione is lobeglitazone. In some embodiments, the signs and symptoms of gastroparesis are measured by ANMS GCSI-DD. In some embodiments, the signs and symptoms of gastroparesis are measured by the change in GCSI-DD. In some embodiments, the signs and symptoms of gastroparesis are measured by the change in GCSI-DD score from baseline to 4 weeks of treatment. In some embodiments, the signs and symptoms of gastroparesis are measured by the change in GCSI-DD score from baseline to 8 weeks of treatment. In some embodiments, signs and symptoms of gastroparesis are measured by the change in GCSI-DD score from baseline to 12 weeks of treatment. In some embodiments, signs and symptoms of gastroparesis are measured by the change in GCSI-DD score from baseline to one year of treatment. In some embodiments, signs and symptoms of gastroparesis are measured by the change in GCSI-DD score from baseline to more than one year of treatment.

[0020] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone to reduce the severity of nausea in subjects with gastroparesis.In some embodiments, the thiazolidinedione is lobeglitazone.In some embodiments, the reduction in the severity of nausea is measured by the change in the severity of nausea from baseline to 4 weeks of treatment.In some embodiments, the reduction in the severity of nausea is measured by the change in the severity of nausea from baseline to 8 weeks of treatment.In some embodiments, the reduction in the severity of nausea is measured by the change in the severity of nausea from baseline to 12 weeks of treatment. In some embodiments, the reduction in severity of nausea is measured by the change in severity of nausea from baseline to one year of treatment.

[0021] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone to reduce the severity of early satiety in subjects with gastroparesis. In some embodiments, the thiazolidinedione is lobeglitazone. In some embodiments, the reduction in the severity of early satiety is measured by the change in the severity of early satiety from baseline to 4 weeks of treatment. In some embodiments, the reduction in the severity of early satiety is measured by the change in the severity of early satiety from baseline to 8 weeks of treatment. In some embodiments, the reduction in the severity of early satiety is measured by the change in the severity of early satiety from baseline to 12 weeks of treatment. In some embodiments, the reduction in severity of early satiety is measured by the change in severity of early satiety from baseline to one year of treatment. In some embodiments, the reduction in severity of early satiety is measured by the change in severity of early satiety from baseline to more than one year of treatment.

[0022] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone to reduce the severity of postprandial fullness in subjects with gastroparesis.In some embodiments, the thiazolidinedione is lobeglitazone.In some embodiments, the reduction in the severity of postprandial fullness is measured by the change in the severity of postprandial fullness from baseline to 4 weeks of treatment.In some embodiments, the reduction in the severity of postprandial fullness is measured by the change in the severity of postprandial fullness from baseline to 8 weeks of treatment.In some embodiments, the reduction in the severity of postprandial fullness is measured by the change in the severity of postprandial fullness from baseline to 12 weeks of treatment. In some embodiments, the reduction in severity of postprandial bloating is measured by the change in severity of postprandial bloating from baseline to one year of treatment. In some embodiments, the reduction in severity of postprandial bloating is measured by the change in severity of postprandial bloating from baseline to more than one year of treatment.

[0023] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone for reducing the severity of upper abdominal pain in subjects with gastroparesis.In some embodiments, the thiazolidinedione is lobeglitazone.In some embodiments, the reduction in the severity of upper abdominal pain is measured by the change in the severity of upper abdominal pain from baseline to 4 weeks of treatment.In some embodiments, the reduction in the severity of upper abdominal pain is measured by the change in the severity of upper abdominal pain from baseline to 8 weeks of treatment.In some embodiments, the reduction in the severity of upper abdominal pain is measured by the change in the severity of upper abdominal pain from baseline to 12 weeks of treatment. In some embodiments, the reduction in severity of upper abdominal pain is measured by the change in severity of upper abdominal pain from baseline to one year of treatment. In some embodiments, the reduction in severity of upper abdominal pain is measured by the change in severity of upper abdominal pain from baseline to more than one year of treatment.

[0024] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone to reduce the frequency of vomiting episodes in subjects with gastroparesis. In some embodiments, the thiazolidinedione is lobeglitazone. In some embodiments, the reduction in the severity of vomiting episodes is measured by the change in the severity of vomiting episodes from baseline to 4 weeks of treatment. In some embodiments, the reduction in the severity of vomiting episodes is measured by the change in the severity of vomiting episodes from baseline to 8 weeks of treatment. In some embodiments, the reduction in the severity of vomiting episodes is measured by the change in the severity of vomiting episodes from baseline to 12 weeks of treatment. In some embodiments, the reduction in severity of emesis episodes is measured by the change in severity of emesis episodes from baseline to one year of treatment. In some embodiments, the reduction in severity of emesis episodes is measured by the change in severity of emesis episodes from baseline to more than one year of treatment.

[0025] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone for reducing the overall severity of gastroparesis. In some embodiments, the thiazolidinedione is lobeglitazone. In some embodiments, the reduction in the severity of the overall severity of gastroparesis is measured by the change in the severity of the overall severity of gastroparesis from baseline to 4 weeks of treatment. In some embodiments, the reduction in the severity of the overall severity of gastroparesis is measured by the change in the severity of the overall severity of gastroparesis from baseline to 8 weeks of treatment. In some embodiments, the reduction in the severity of the overall severity of gastroparesis is measured by the change in the severity of the overall severity of gastroparesis from baseline to 12 weeks of treatment. In some embodiments, the reduction in severity of the general severity of gastroparesis is measured by the change in severity of the general severity of gastroparesis from baseline to one year of treatment. In some embodiments, the reduction in severity of the general severity of gastroparesis is measured by the change in severity of the general severity of gastroparesis from baseline to more than one year of treatment.

[0026] Also disclosed herein is the use of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone to slow, halt, or reverse disease progression of GI disorders. In some embodiments, the thiazolidinedione is lobeglitazone.

[0027] Also disclosed herein is a method for treating GI disorders in a mammal, comprising administering to the mammal an effective amount of a thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, and balaglitazone.In some embodiments, the GI disorder is gastroparesis or IBD.In some embodiments, the thiazolidinedione is lobeglitazone.

[0028] In some embodiments, the gastroparesis is idiopathic gastroparesis, diabetic gastroparesis, post-operative gastroparesis, or medication-induced gastroparesis.

[0029] Also disclosed herein is a method of inhibiting M1 macrophage differentiation in an individual, comprising administering to the individual an effective amount of lobeglitazone. In some embodiments, as a result of inhibiting M1 macrophage differentiation, a GI system disorder is treated in the individual.

[0030] Also disclosed herein is a method of promoting M1 macrophage differentiation in an individual, comprising administering to the individual an effective amount of pioglitazone or rosiglitazone.

[0031] Also disclosed herein is a method of promoting M2 macrophage differentiation in an individual, comprising administering an effective amount of lobeglitazone, pioglitazone, or rosiglitazone to the individual. In some embodiments, as a result of the promotion of M2 macrophage differentiation, a GI system disorder is treated for the individual. Also disclosed herein is a method of promoting repolarization of M1 macrophages to M2 macrophages in an individual, comprising administering an effective amount of lobeglitazone, pioglitazone, or rosiglitazone to the individual. In some embodiments, as a result of the promotion of repolarization of M1 macrophages to M2 macrophages, a GI system disorder is treated for the individual. In some embodiments, the thiazolidinedione is lobeglitazone. In some embodiments, the GI disorder is selected from abdominal adhesions, adult acid reflux (GERD), pediatric acid reflux (GERD), anatomical problems of the lower GI tract, appendicitis, Barrett's esophagus, bowel control problems (fecal incontinence), celiac disease, colonic polyps, constipation, Crohn's disease, cyclic vomiting, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, intestinal pseudo-obstruction, IBD, IBS, lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcers (gastric ulcers), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, and Zollinger-Ellison syndrome, or symptoms associated therewith. [Brief description of the drawings]

[0032] [Figure 1] FIG. 1 represents the experimental timeline for lobeglitazone studies in a mouse model of inflammatory bowel disease (IBD). [Diagram 2] Figure 2 shows the body weights of animals in the untreated group, 5% dextran sulfate sodium (DSS) model group (model), lobeglitazone group, and cyclosporine A group (mpk indicates mg / kg). Compared to the model control group, * indicates p<0.05, and ** indicates p<0.01. [Diagram 3]3 shows the changes in body weight of animals in the untreated group, 5% DSS model group, lobeglitazone group, and cyclosporine A group. Compared to the model control group, * indicates p<0.05, and ** indicates p<0.01. [Figure 4] 4 shows the Disease Activity Index (DAI) scores for the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. Compared to the model control group, ** indicates p<0.01, and *** indicates p<0.001. [Diagram 5] FIG. 5 shows colon weights after treatment in the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. [Figure 6] 6 shows colon length after treatment in the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. Compared to the model control group, * indicates p<0.05, and *** indicates p<0.001. [Figure 7] 7 shows the colon weight / length ratio after treatment in the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. Compared to the model control group, * indicates p<0.05. [Figure 8] 8 shows TNF-α gene expression after treatment in the colon of the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. Compared to the model control group, * indicates p<0.05, and *** indicates p<0.001. [Figure 9] FIG. 9 shows IL-10 gene expression after treatment in the colon of the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. [Figure 10] 10 shows the expression of CCL2 gene after treatment in the colon of the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. Compared to the model control group, * indicates p<0.05, and ** indicates p<0.01. [Figure 11] 11 shows IL-1β gene expression after treatment in the colon of the untreated group, the 5% DSS model group, the lobeglitazone group, and the cyclosporine A group. Compared to the model control group, *** is p<0.001. [Figure 12]Figure 12 shows the inflammation / tissue damage scores (total hematoxylin and eosin (H&E) scores) after treatment for the untreated group, 5% DSS model group, lobeglitazone group, and cyclosporine A group. Compared to the model control group, ** is p<0.01, ** is p<0.001. [Figure 13] Figure 13 shows the number of goblet cells (Periodic Acid Schiff (PAS) staining) after treatment in the untreated group, 5% DSS model group, lobeglitazone group, and cyclosporine A group. Compared to the model control group, * is p<0.05. [Figure 14] FIG. 14 depicts M1 macrophage cytokine IL-23 expression following differentiation with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 15] FIG. 15 depicts M1 macrophage cytokine IL-1b expression following differentiation with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 16] FIG. 16 depicts M1-like macrophage cytokine TNF-alpha expression following differentiation with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 17] FIG. 17 depicts M1-like macrophage cytokine IL-12p70 expression following differentiation with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 18] FIG. 18 depicts M1-like macrophage cytokine IL-6 expression following differentiation with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 19] FIG. 19 depicts M2 macrophage surface marker expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone (donor 1). [Figure 20] FIG. 20 depicts M2 macrophage surface marker expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone (donor 2). [Figure 21] FIG. 21 depicts M2 macrophage surface marker expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone (donor 3). [Figure 22] FIG. 22 depicts M1 macrophage-associated cytokine expression in cells from three donors following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Diagram 23] FIG. 23 depicts M2 macrophage-associated cytokine expression in cells from three donors following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 24] FIG. 24 depicts post-treatment macrophage-derived chemokine (MDC) expression in cells from three donors following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Diagram 25] FIG. 25 depicts post-treatment eotaxin-2 expression in cells from three donors following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 26] FIG. 26 depicts post-treatment IL-1RA expression in cells from three donors following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 27] FIG. 27 depicts M2 macrophage surface marker expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone (donor 1). [Figure 28] FIG. 28 depicts M1 macrophage-associated cytokine expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Figure 29] FIG. 29 depicts M2 macrophage-associated cytokine expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Diagram 30] FIG. 30 depicts MDC expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Diagram 31] FIG. 31 depicts eotaxin-2 expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone. [Diagram 32] FIG. 32 depicts IL-1RA expression following treatment with lobeglitazone, pioglitazone, or rosiglitazone. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0033] Disclosed herein are methods of treating a GI disease or disorder with a thiazolidinedione disclosed herein.

[0034] The term "thiazolidinedione" refers to a group of heterocyclic glitazone compounds containing a five-membered C3NS ring, including their prodrugs, salts, solvates, hydrates, cocrystals, enantiomers, and deuterated forms. As used herein, thiazolidinediones include, but are not limited to, one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, balaglitazone, and other thiazolidinedione molecules. In some embodiments disclosed herein, the method includes using any thiazolidinedione. In some embodiments, the thiazolidinedione is lobeglitazone. In some embodiments, the method specifically excludes the use of one or more thiazolidinediones disclosed herein. In some embodiments, the thiazolidinedione is not pioglitazone. In some embodiments, the thiazolidinedione is not pioglitazone or rosiglitazone.

[0035] Lobeglitazone (Duvie®, Chong Kun Dang) is a thiazolidinedione with the chemical name 5-(4-(2-((6-(4-methoxyphenoxy)pyrimidin-4-yl)(methyl)amino)ethoxy)benzyl)thiazolidine-2,4-dione. As an agonist of both peroxisome proliferator-activated receptors alpha (PPARα) and gamma (PPARγ), lobeglitazone binds to PPAR receptors in fat cells and acts as an insulin sensitizer by making the cells more responsive to insulin.

[0036] Pioglitazone (Actos®, Takeda) is a thiazolidinedione with the chemical name (±)-5-[p-[2-(5-ethyl-2-pyridyl)ethoxy]benzyl]-2,4-thiazolidinedione monohydrochloride. Pioglitazone is a potent and highly selective agonist for PPARγ, improving sensitivity to insulin in muscle and adipose tissue and inhibiting hepatic gluconeogenesis. Pioglitazone reduces circulating insulin concentrations while improving glycemic control.

[0037] Rosiglitazone (Avandia®, GlaxoSmithKline) is a thiazolidinedione with the chemical name (RS)-5-[4-(2-[methyl(pyridin-2-yl)amino]ethoxy)benzyl]thiazolidine-2,4-dione. Like other thiazolidinediones, the mechanism of action of rosiglitazone is through activation of intracellular PPARs, specifically PPARγ. Rosiglitazone is a selective ligand for PPARγ and has no PPARα binding activity.

[0038] As used herein, the terms "treat," "treating," or "treatment" mean to alleviate, reduce, or abolish one or more symptoms or characteristics of a disease, and may cure, palliate, prevent, or slow the progression of a disease.

[0039] The term "effective amount" refers to an amount that reduces the corresponding or specified damage to the digestive system and produces the desired effect or result. The term "therapeutically effective amount" refers to an amount of thiazolidinedione, including but not limited to one or more of pioglitazone, rosiglitazone, lobeglitazone, ciglitazone, darglitazone, deuterium-stabilized R-pioglitazone, englitazone, leriglitazone, netoglitazone, rivoglitazone, troglitazone, balaglitazone, and other thiazolidinedione molecules, that is effective, alone or in combination with other active ingredients, to induce the desired biological or pharmacological response, for example, to prevent, alleviate, or improve the symptoms of a disease or disorder; to slow, stop, or reverse the course or progression of an underlying disease; to partially or completely restore cellular function; or to prolong the survival of the subject being treated. In some embodiments, the thiazolidinedione is not pioglitazone.

[0040] The term "patient" or "subject" includes mammals, including non-human mammals and humans. In one embodiment, the patient or subject is a human.

[0041] The terms "significant" or "significantly" are determined by a t-test at a significance level of 0.05.

[0042] The method is anticipated based on the surprising finding that thiazolidinediones can significantly protect the digestive system from damage as evidenced by inflammation. Thus, disclosed herein is the use of one or more thiazolidinediones for the treatment of GI disorders.

[0043] As used herein, the terms "digestive" and "digestive tract" are used interchangeably and refer to the organs, cells, tissues, and diseases associated with the gastrointestinal (GI) tract, including, but not limited to, the mouth, pharynx (throat), esophagus, stomach, small intestine, large intestine, rectum, and anus, along with the salivary glands, liver, pancreas, and gallbladder.

[0044] Digestive disorders (gastrointestinal disorders) caused by damage to the digestive system include abdominal adhesions, acid reflux in adults (gastroesophageal reflux disease or GERD), acid reflux in children (GERD), anatomical problems of the lower GI tract, appendicitis, Barrett's esophagus, bowel management problems (fecal incontinence), celiac disease, colon polyps, constipation, Crohn's disease, cyclic vomiting, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gastric cancer, gastrointestinal problems, These conditions include, but are not limited to, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, intestinal pseudo-obstruction, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcer (stomach ulcer), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, Zollinger-Ellison syndrome, or conditions associated with these.

[0045] Intestinal macrophages are the primary immune cells in the maintenance of intestinal immune homeostasis and have a role in the pathogenesis of GI diseases such as inflammatory bowel disease. According to microenvironmental cues, macrophages polarize into two distinct phenotypes: classically activated (M1) or alternatively activated (M2) macrophages. For example, when intestinal barrier function is compromised, efferocytosis effects result in macrophages functionally polarizing to the M2 subtype, producing cytokines, chemokines, and lipid mediators involved in healing the intestinal mucosal barrier and maintaining homeostasis. Various cytokines and other soluble factors, including prostaglandin E2 (PGE2), bone morphogenetic protein 2 (BMP2), and WNT ligands, can be produced by intestinal macrophages that stimulate the expansion of epithelial cells and participate in the enteric nervous system or the intestinal mucosal barrier to help maintain tissue homeostasis. Intestinal macrophages inhibit antigen-specific CD4 expression, primarily by producing IL-10, to inhibit uncontrolled inflammation in response to harmless commensal microorganisms. + CD25 + Promotes the proliferation of regulatory T (T-reg) cells to maintain tolerance.

[0046] Macrophages infiltration and activation can phagocytose pathogens, as well as produce various cytokines under certain circumstances, and cooperate with distinct immune cells in many aspects of the pathogenesis of GI diseases. Imbalance in macrophage polarization results in the exacerbation of GI diseases, and the production of certain cytokines and / or chemokines depends on the ratio of proinflammatory M1 subset to anti-inflammatory M2 subset. Therefore, targeted therapy of macrophages is a novel option to adjust immune microenvironment and remodel intestinal tissue.

[0047] Gastroparesis is a disorder characterized by delayed gastric emptying (DGE) in the absence of mechanical obstruction. Symptoms are chronic, with episodic exacerbations. The idiopathic form of the disorder, which accounts for the majority of cases, mainly affects young adult women. Gastroparesis is also often associated with diabetes (diabetic gastroparesis), which likely occurs due to impaired neural control of gastric motility. Macrophages play a role in the development and progression of gastroparesis. In particular, macrophages infiltrate the smooth muscle layer of the stomach in people with gastroparesis and release proinflammatory molecules that contribute to abdominal muscle damage and dysfunction. There are two distinct subtypes of macrophages that have various functions in the immune system: M1 macrophages and M2 macrophages. M1 macrophages are involved in the inflammatory response and are activated in response to proinflammatory signals. M2 macrophages produce anti-inflammatory cytokines and growth factors that help repair damaged tissue and promote angiogenesis. Overall, M1 and M2 macrophages represent two different functional states of macrophages, allowing them to respond to different signals and play different roles in the immune system. The balance between M1 and M2 macrophages is important in maintaining immune homeostasis and promoting appropriate immune responses. Drugs that can inhibit M1 macrophages, promote M2 macrophages, and / or allow repolarization from M1 macrophages to M2 macrophages may be able to treat gastrointestinal diseases, including gastroparesis.

[0048] Thus, the present disclosure also provides for the use of a thiazolidinedione in inhibiting or promoting M1 macrophage differentiation and / or promoting M2 macrophage differentiation. In some embodiments, the thiazolidinedione is lobeglitazone.

[0049] The signs and symptoms of gastroparesis are nausea, vomiting, postprandial bloating, early satiety, and epigastric pain. Patients may experience any combination of signs and symptoms of varying degrees of severity. Pain is more prevalent in patients with idiopathic gastroparesis than in patients with diabetic gastroparesis. Patients with diabetic gastroparesis may also experience disturbances in glycemic control due to unpredictable gastric emptying and altered absorption of orally administered hypoglycemic drugs.

[0050] Understanding the associated symptoms of gastroparesis is important in treating patients with this disorder. In gastroparesis, the experience and severity of symptoms are derived from the patient. As a result, a patient-reported symptom scale that captures the overall severity of gastroparesis is necessary to evaluate treatments for gastroparesis. A clearly defined patient-reported outcome (PRO) instrument that measures clinically significant signs and symptoms of gastroparesis would be a useful evaluation tool for clinical trials to support labeling claims for treatments for gastroparesis. The American Society of Neurogastroenterology and Motility Gastroparesis Predominant Symptom Index Diary (ANMS GCSI-DD) is a patient-reported outcome instrument that captures the daily associated symptoms of gastroparesis. The ANMS GCSI-DD is designed to assess GI symptoms associated with idiopathic gastroparesis and diabetic gastroparesis. The ANMS GCSI-DD is designed to assess symptoms associated with idiopathic gastroparesis and diabetic gastroparesis. The ANMS GCSI-DD includes five items: nausea, vomiting, early satiety, postprandial fullness, and epigastric pain. Four of these items (nausea, early satiety, postprandial fullness, and epigastric pain) are rated on a scale of none (0), mild (1), moderate (2), severe (3), and severe (4) based on the worst severity of symptoms over the past 24 hours. Vomiting is rated as the number of vomiting episodes over the past 24 hours, with the maximum number limited to four (range 0–4). The ANMS GCSI-DD total score is a patient-reported outcome useful for gastroparesis and as an endpoint in clinical trials of gastroparesis.

[0051] In certain aspects, the disclosure provides a method of exerting a protective effect in a cell, the method comprising contacting the cell with an effective amount of a thiazolidinedione. As used herein, the term "effective amount" refers to an amount of a thiazolidinedione that results in a desired effect or result, e.g., an amount that results in a protective effect.

[0052] In certain aspects, the disclosure provides a method of reducing inflammation comprising contacting a cell with an effective amount of a thiazolidinedione.

[0053] In certain aspects, the disclosure provides a method of extending the lifespan of a cell, the method comprising contacting the cell with an effective amount of a thiazolidinedione.

[0054] In some embodiments, the cell is an animal cell, e.g., a mammalian cell. In some embodiments, the cell is a human cell or a non-human cell. In some embodiments, the cell is treated in vitro, in vivo, or ex vivo. In some embodiments, the cell is a diseased cell. In some embodiments, the cell is a diseased cell from a patient suffering from a disease or disorder disclosed herein.

[0055] Also disclosed herein is a method of treating a mammal having a disease or disorder, to benefit from a protective effect on cells or to prevent or reduce the risk of acquiring the disease or disorder in the mammal, comprising administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a thiazolidinedione. In some embodiments, the mammal is a human or non-human mammal. In some embodiments, the mammal is a human. In some embodiments, the disease or disorder is caused by an injury that reduces the function of the digestive system. In some embodiments, the disease is selected from one or more digestive diseases or symptoms associated therewith. In some embodiments, the disease is gastroparesis. In some embodiments, the disease is idiopathic gastroparesis, diabetic gastroparesis, or mediation-induced gastroparesis.

[0056] Also disclosed herein is a method for treating a disease or disorder, which method extends the lifespan of cells in a patient having a disease or disorder that results in damage or inflammation of the digestive system in a mammal, or that results in damage to the digestive system.

[0057] In some embodiments, the method includes administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising a thiazolidinedione. In some embodiments, the mammal is a human or a non-human mammal. In one embodiment, the disease or disorder is selected from, but is not limited to, abdominal adhesions, acid reflux in adults (gastroesophageal reflux disease or GERD), acid reflux in children (GERD), anatomical problems of the lower GI tract, appendicitis, Barrett's esophagus, bowel control problems (fecal incontinence), celiac disease, colonic polyps, constipation, Crohn's disease, cyclic vomiting, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, pseudo-obstruction, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcers (gastric ulcers), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, Zollinger-Ellison syndrome, or symptoms associated therewith.

[0058] Also disclosed herein is a method for treating gastroparesis.In some embodiments, gastroparesis includes idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis and drug-induced gastroparesis.In some embodiments, primary gastroparesis is idiopathic.In some embodiments, secondary gastroparesis is caused by disease such as diabetes, cancer or infection, or by drug side effects, or by surgery.

[0059] In another embodiment, provided herein is a method of reducing inflammation in the GI tract for the treatment of gastrointestinal disorders such as gastroparesis and IBD.

[0060] In another aspect, provided herein is a method of reducing the risk of nausea, early satiety, postprandial bloating, epigastric pain, episodes of vomiting, and the general severity of gastroparesis.

[0061] In another aspect, provided herein is a method of slowing, halting, or reversing disease progression to gastrointestinal disorders such as gastroparesis and IBD.

[0062] In another aspect, provided herein is a method of slowing, halting, or reversing the progression of the disease as indicated by nausea, early satiety, postprandial bloating, epigastric pain, episodes of vomiting, and general severity of gastroparesis.

[0063] In another aspect, disclosed herein is a method of treating a mammal having a disease or disorder whose condition is prevented, alleviated, or ameliorated by cell protection, or whose disease process or progression is slowed, halted, or reversed by cell protection, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a thiazolidinedione. In some embodiments, the thiazolidinedione is lobeglitazone.

[0064] In a related aspect, disclosed herein is a method of treating gastroparesis, which in one embodiment includes idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis, and pharmaceutical-induced gastroparesis.

[0065] The present disclosure further provides the use of a thiazolidinedione for the preparation of a medicament for treating a human having any one of the diseases or disorders disclosed herein, or for use in any method of the disclosure involving administration of a thiazolidinedione to a human.

[0066] The pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of thiazolidinedione and at least one pharma- ceutically acceptable excipient. The term "excipient" refers to a pharma-ceutically acceptable inactive substance used as a carrier for the pharma-ceutically active ingredient thiazolidinedione, and includes anti-adherents, binders, coatings, disintegrants, fillers, diluents, solvents, flavorings, bulkants, colorants, glidants, dispersants, wetting agents, lubricants, preservatives, adsorbents and sweeteners. The choice of excipient will depend on factors such as the specific administration method and nature of the dosage form. The solutions or suspensions used for injection or infusion may contain the following components: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycol, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetic acid, citric acid or phosphate, and agents for adjusting tonicity such as sodium chloride or dextrose. pH may be adjusted using acids or bases such as hydrochloric acid or sodium hydroxide. Parenteral preparations may be enclosed in ampoules, disposable syringes, including autoinjectors, or multi-dose vials made of glass or plastic.

[0067] The pharmaceutical formulation of the present disclosure may be in any pharmaceutical dosage form. The pharmaceutical formulation may be, for example, a tablet, a capsule, a nanoparticle material, e.g., a granular particle material or powder, a lyophilizate for reconstitution, a liquid solution, a suspension, an emulsion or other liquid form, a suspension, a solution, an emulsion, etc. for injection, a suppository, or a topical or transdermal formulation or patch. The pharmaceutical formulation generally comprises about 1% to about 99% by weight of the thiazolidinedione and 99% to 1% by weight of suitable excipients. In one embodiment, the dosage form is an oral dosage form. In another embodiment, the dosage form is a parenteral dosage form. In another embodiment, the dosage form is an enteral dosage form. In another embodiment, the dosage form is a topical dosage form. In one embodiment, the pharmaceutical dosage form is a unit dose. The term "unit dose" refers to the amount of thiazolidinedione administered to a patient in a single dose.

[0068] In some embodiments, the pharmaceutical compositions disclosed herein are delivered to a subject by parenteral, enteral, or topical routes.

[0069] Examples of parental routes suitable for use with the disclosed pharmaceutical compositions include the following: intraperitoneal, intraamniotic, intra-arterial, intra-articular, intrabiliary, intra-bronchial, intra-vesical, intracardiac, intracartilaginous, intrasacral, intracavity, intracavity, intracerebral, intracisternal, intracorneal, intracoronary, intracoronary, intracavernous, intracranial, intradermal, intraspinal, intraductal, intraduodenal, intradural, intraepidermal, intraesophageal, intragastric, intragingival, intraileal, intralesional, intraluminal, pulmonary, intraluminal ... The administration route may include, but is not limited to, any one or more of intraglandular, intramedullary, intrameningeal, intramuscular, intraocular, intraovarian, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intraocular, intrasinusoidal, intraspinal, intrasynovial, intratendinous, intratesticular, intraspinal, intrathoracic, intratubular, intratumoral, intratympanic, intrauterine, intravascular, intravenous (bolus or infusion), intracerebral, intravesical, and / or subcutaneous. In some embodiments, the administration route is not buccal or sublingual. In some embodiments, the administration route is not nasal.

[0070] Enteral routes of administration include administration to the GI tract via the mouth (oral), stomach (intragastric), and rectum (intragastric). Intragastric administration typically involves the use of a tube through the nasal passages (NG tube) or a tube in the esophagus directly into the stomach (PEG tube). Rectal administration typically requires a rectal suppository.

[0071] Topical administration, including pulmonary administration, includes administration to a body surface, such as the skin or mucous membranes. Transdermal forms include creams, foams, gels, lotions or ointments. Pulmonary forms include solutions and powders, such as liquid sprays.

[0072] The dosage can vary depending on the dosage form used, the sensitivity of the patient, and the route of administration. The dosage is adjusted to give sufficient levels of the active agent or to maintain the desired effect. Factors to be considered include the severity of the condition, the general health of the subject, the age, weight, and sex of the subject, diet, number and frequency of administration, drug combinations, reaction sensitivities, and tolerance / response to treatment.

[0073] In one embodiment, the daily amount of thiazolidinedione administered to the patient is selected from up to 200 mg, 175 mg, 150 mg, 125 mg, 100 mg, 90 mg, 80 mg, 70 mg, 60 mg, 50 mg, 30 mg, 25 mg, 20 mg, 15 mg, 14 mg, 13 mg, 12 mg, 11 mg, 10 mg, 9 mg, 8 mg, 7 mg, 6 mg, 5 mg, 4 mg, 3 mg, 2 mg, 1 mg, 0.9 mg, 0.8 mg, 0.7 mg, 0.6 mg, 0.5 mg, 0.45 mg, 0.4 mg, 0.3 mg, 0.2 mg, 0.1 mg, 0.08 mg, 0.05 mg, 0.03 mg, 0.02 mg or up to 0.01 mg. In another embodiment, the daily dose is at least 0.01 mg, 0.02 mg, 0.05 mg, 0.08 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.45 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 12 mg, 13 mg, 14 mg, 15 mg g, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1,000 mg, 2,000 mg, 3,000 mg, 4,000 mg, or at least 5,000 mg.In another embodiment, the daily dose is 0.01-0.0.2 mg, 0.02-0.05 mg, 0.05-0.08 mg, 0.08-0.1 mg, 0.1-0.2 mg, 0.2-0.4 mg, 0.4-0.6 mg, 0.6-0.8 mg, 0.8-1 mg, 1-2 mg, 2-4 mg, 1-5 mg, 5-7.5 mg, 7.5-10 mg, 10-15 mg, 10-12.5 mg, 12.5-15 mg, 15-17.7 mg, 17.5-20 mg, 20-25 mg, 20-22.5 mg, 22.5-25 mg, 25-30 mg, 25-27.5 mg, 26-30 mg, 27-31 mg, 28-32 mg, 29-40 mg, 30-33 mg, 31-34 mg, 32-35 mg, 33-36 mg, 34-37 mg, 35-38 mg, 36-39 mg, 37-40 mg, 38-41 mg, 38-42 mg, 39-43 mg, 40-44 mg, 41-45 mg, 42-46 mg, 43-47 mg, 44-48 mg, 45-49 mg, 46-50 mg, 47-51 mg, 48-52 mg, 49-53 mg, 54-55 mg, 55-56 mg, 56-57 mg, 57-58 mg, 58-59 mg, 59-60 mg, 60-61 mg, 61-62 mg, 62-63 mg, 63-64 mg, 27.5-30 mg, 30-35 mg, 35-40 mg, 40-45 mg, or 45-50 mg, 50-75 mg, 75-100 mg, 100-125 mg, 125-150 mg, 150-175 mg, 175-200 mg, 5-200 mg, 5-300 mg, 5-400 mg, 5-500 mg, 5-600 mg, 5-700 mg, 5-800 mg, 5-900 mg, 5-1,000 mg, 5-2,000 mg, 5-5,000 mg, or greater than 5,000 mg, or any range bounded by a pair of these values.

[0074] In another embodiment, The single dose of thiazolidinedione administered to patients is 0.01mg, 0.02mg, 0.05mg, 0.08mg, 0.1mg, 0.2mg, 0.3mg, 0.4mg, 0.45mg, 0.5mg, 0.6mg, 0.7mg, 0.8mg, 0.9mg, 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg, 9mg, 10mg, 12mg, 13mg, 14mg, 15mg, 16mg, 17mg, 18mg, 19mg, 20mg, 21mg, 22mg, 23mg, 24mg, 25mg, 26mg, 27mg, 28mg, 29mg, 30mg, 35mg, 40mg, 45mg, 50mg, 100mg, 110mg, 120mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 350mg, 360mg, 370mg, 380mg, 390mg, 400mg, 410mg, 420mg, 430mg, 440mg, 450mg, 460mg, 470mg, 480mg, 490mg, 500mg, 500mg, 510mg, 520mg, 530mg, 540 In some embodiments, the compound is selected from 0 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, 450 mg, 460 mg, 470 mg, 480 mg, 490 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1,000 mg, 2,000 mg, 3,000 mg, 4,000 mg, or 5,000 mg, or any range bounded by a pair of these values. In one embodiment, the single dose is administered by oral administration. In another embodiment, the single dose is administered by injection, e.g., subcutaneous injection, intramuscular injection, or intravenous injection. In another embodiment, the single dose is administered by inhalation administration. In some embodiments, the thiazolidinedione is lobeglitazone.

[0075] As a non-limiting example, the dosage of a thiazolidinedione, such as lobeglitazone, administered by oral administration is about 0.01-50 mg per day, which may be administered in divided doses. A single dose of a thiazolidinedione, such as lobeglitazone, administered by subcutaneous injection may be about 0.01-50 mg, preferably about 0.1-10 mg, 0.2-1 mg, 0.3-0.6 mg, or 0.5 mg, or any range bounded by these value pairs. Other embodiments include a range of about 0.05-5,000 mg, preferably about 0.1-10 mg, 0.2-5 mg, 0.3-1 mg, or 0.5 mg, or any range bounded by these value pairs. For those patients who require more than 10 divided injections per day, subcutaneous infusion may be preferred.

[0076] A fine particle dose of a thiazolidinedione such as lobeglitazone administered by pulmonary administration, e.g., by inhalation using a pressurized metered dose inhaler (pMDI), dry powder inhaler (DPI), soft mist inhaler, nebulizer, or other device, can be in the range of about 0.1 to 50 mg, preferably about 0.2 to 10 mg, 0.3 to 1 mg, or 0.5 mg, or any range bounded by these value pairs. Other embodiments include in the range of about 0.05 to 5,000 mg, preferably about 0.1 to 1,000 mg, 0.2 to 100 mg, 0.3 to 1 mg, 0.4 to 0.5 mg, or 0.5 mg, or any range bounded by these value pairs. The nominal dose (ND), i.e., the amount of drug metered into a container of a thiazolidinedione such as lobeglitazone administered by pulmonary administration (also known as a metered dose inhaler), can be, for example, within the range of 0.1-15 mg, 0.1-10 mg, 0.1-1 mg, 0.2-0.3 mg, 0.3-0.4 mg, 0.4-0.5 mg, 0.5-0.6 mg, 0.6-0.7 mg, 0.7-0.8 mg, 0.8-0.9 mg, or 0.9-1 mg, or any range bounded by these value pairs. Other embodiments include the range of about 0.05-5,000 mg, preferably about 0.1-1,000 mg, 0.2-10 mg, 0.3-1 mg, 0.4-0.5 mg, or 0.5 mg, or any range bounded by these value pairs.

[0077] Long-acting pharmaceutical compositions may be administered once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times or more than ten times per day (preferably ≦10 times per day), every other day, every 3-4 days, every week, or once every two weeks depending on the half-life and clearance rate of the particular formulation.

[0078] In one embodiment of any of the above methods and compositions, the thiazolidinedione, or salts, solvates, hydrates, and cocrystals thereof, is a racemic mixture of the R and S enantiomers, or is R-enantiomer-rich (i.e., the ratio of R to S enantiomers administered is between 1.1:1 and 1,000:1, 10:1 and 10,000:1, or 100:1 and 100,000:1, or the total of all thiazolidinedione enantiomers in the composition is at least 98% R enantiomer, 99% S enantiomer, 99.5% thiazolidinedione enantiomer, or 100% S enantiomer). or is S enantiomer-rich (i.e., the ratio of S enantiomer to R enantiomer is between 1.1:1 and 1,000:1, 10:1 and 10,000:1, or 100:1 and 100,000:1, or all of the thiazolidinedione enantiomers in the composition as a whole are at least 98% S enantiomer, 99% enantiomer, 99.5% enantiomer, 99.9% enantiomer, or contain no detectable amount of R enantiomer).

[0079] The present disclosure further provides an in vitro or ex vivo method of reducing cell damage comprising contacting a cell with an effective amount of a thiazolidinedione.

[0080] Suitably, the cells are used to treat conditions such as abdominal adhesions, acid reflux in adults (gastroesophageal reflux disease or GERD), acid reflux in children (GERD), anatomical problems of the lower GI tract, appendicitis, Barrett's esophagus, bowel management problems (fecal incontinence), celiac disease, colon polyps, constipation, Crohn's disease, cyclic vomiting, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, pseudo-obstruction. have or be at risk for or be at risk of acquiring a disease or disorder selected from any one or more of: inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcer (stomach ulcer), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, Zollinger-Ellison syndrome, or symptoms associated therewith.

[0081] Preferably, in the methods, compositions and / or second medical uses of the presently disclosed compositions, the thiazolidinedione is administered or formulated for administration in a dosage of 0.01 mg or more. Preferably, the thiazolidinedione is administered in a dosage of 0.1 to 5000 mg per day.

[0082] Suitably, in the methods, compositions and / or secondary medical uses of the presently disclosed compositions, the thiazolidinedione may be administered or formulated for administration in any suitable manner, such as parenterally, enterally, or topically.

[0083] Suitably, in the methods, compositions and / or secondary medical uses of the presently disclosed compositions, the thiazolidinedione may be administered or formulated for administration by oral, sublingual, buccal, pulmonary, intravenous, intramuscular, or subcutaneous administration.

[0084] Another embodiment of the present disclosure includes the use of thiazolidinediones to reduce cell damage or improve cell survival.

[0085] Another embodiment of the present disclosure includes the use of thiazolidinediones to treat gastroparesis or IBD.

[0086] Another embodiment of the present disclosure includes the use of thiazolidinediones to reduce the core signs and symptoms of gastroparesis as measured by the ANMS GCSI-DD.

[0087] Another embodiment of the present disclosure includes the use of a thiazolidinedione to reduce the severity of nausea.

[0088] Another embodiment of the present disclosure includes the use of a thiazolidinedione to reduce the severity of early satiety.

[0089] Another embodiment of the present disclosure includes the use of a thiazolidinedione to reduce the severity of post-prandial bloating.

[0090] Another embodiment of the present disclosure includes the use of a thiazolidinedione to reduce the severity of upper abdominal pain.

[0091] Another embodiment of the present disclosure involves the use of thiazolidinediones to reduce the severity of vomiting episodes.

[0092] Another embodiment of the present disclosure includes the use of a thiazolidinedione to reduce the overall severity of gastroparesis.

[0093] Working Example The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will be apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0094] It should be further understood that all values ​​are approximate and are given for illustrative purposes. All references cited and discussed herein are incorporated by reference in their entirety and to the same extent as if each reference was individually incorporated by reference.

[0095] Example 1: Efficacy of Lobeglitazone to Treat GI Inflammation procedure

[0096] The aim of this study was to test the efficacy of the compound lobeglitazone in a DSS-induced IBD model in female Balb / c mice after oral administration. The study was performed in an AAALAC-accredited facility and all animal experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC).

[0097] This study was carried out according to the procedures outlined below.

[0098] Preparation of vehicle and test articles

[0099] A total of 5 g of carboxymethylcellulose (CMC) powder was weighed and transferred into a clean bottle, followed by the addition of 1 L of Milli-Q water. A stir bar was then added and the solution was stirred for 6 hours until clear. This 0.5% CMC was stored at 4°C for further use.

[0100] A precise amount of lobeglitazone sulfate (Lot: EW34258-13-P1; purity: 97.6%) was weighed into a clean tube, 0.5% CMC was added, vortexed thoroughly, and then sonicated for 2-3 min to obtain a homogenous suspension. The concentrations of the solutions were 0.361 mg / mL, 0.120 mg / mL, and 0.036 mg / mL. Dosing solutions were made up daily until the end of the study. The compound was administered orally (PO) at 10 mL / kg daily for 10 days.

[0101] A total of 125.82 mg of cyclosporine A (CSA, 98% purity) was weighed into a clean tube, 2.055 mL of castor oil was added first, then 39.046 mL of 95% CMC (containing 0.1% tween 80) was added, vortexed thoroughly, and then sonicated for 10 minutes to obtain a clear solution. The concentration of the solution was 3 mg / mL. 10 mL / kg of compound was administered PO daily for 10 days.

[0102] 5% DSS preparation: 5 g of DSS powder was weighed into a clean bottle, followed by the addition of 1 L of Milli-Q water and stirring the solution until clear. The 5% DSS was then transferred to the animals' water bottles. The 5% DSS was made up daily until the end of the study.

[0103] Animal husbandry

[0104] A total of 50 female Balb / c (7 weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The animals were acclimated for one week prior to the experiment. All in vivo experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC). All euthanasia was performed using carbon dioxide inhalation, and every effort was made to minimize animal suffering.

[0105] Group and dose design

[0106] Randomization was performed based on body weight at the start of the study. All animals were randomly assigned to one of six groups (Table 1). The study schedule is described in Figure 1.

[0107] [Table 1]

[0108] Animal Health Monitoring

[0109] During the study, the general condition (appearance and activity) of all animals was closely monitored daily by a veterinarian.

[0110] Disease Activity Index (DAI) score

[0111] DAI was assessed using the criteria listed in Table 2. The DAI score was calculated as the sum of the weight loss score, stool consistency score, and occult-bleeding score. Weight loss was calculated relative to the body weight on day 1. Fecal samples were collected from each animal and assessed for stool consistency and occult-bleeding according to Table 2. Stool consistency was assessed from the appearance of the stool. Occult-bleeding was assessed using fecal occult blood test strips. If a light or dark purple color develops within 20 seconds and lasts longer than 15 seconds, an occult-bleeding score of 1 is assigned. If a light or dark purple color develops within 15 seconds, an occult-bleeding score of 2 is assigned. If blood is seen in the stool, an occult-bleeding score of 3 is assigned.

[0112] [Table 2]

[0113] Sampling

[0114] At the end of the study, CO 2 The animals were euthanized at 0 °C. Colon length and colon weight (not including the cecum) were recorded and gross changes were scored based on Table 2. Physiological saline was used to wash the colon and all feces were removed before weight measurement. The middle part of the colon was collected in a cassette and fixed in 10% neutral buffered formalin (NBF) for H&E and PAS staining. For the left colon, the mucosa from the colon surface was removed and then snap frozen in liquid nitrogen and stored at -80 °C for QT-PCR analysis of TNF-α, IL-10, CCL2, and IL-1β. QT-PCR primers were purchased from Sangon Biotech Co.,Ltd.

[0115] Treatment of tissues prior to RNA extraction was as follows. (a) Remove the mucosa from the colon surface. Cut the colon tissue into small pieces on ice. (b) Weigh out 30 mg or less of tissue and transfer the tissue to a tube containing lysing matrix D. Immediately add a total of 700 μL of Trizol to the tissue and lysing matrix D. (c) Place the lysing matrix D tube containing the tissue, lysing matrix, and 700 μL of Trizol into the FastPrep™-24 Sample Preparation System. (d) Set to speed 6 and grind for 2 minutes. (e) Add 140 μL of chloroform to the tube and vigorously shake the tube by vortexing for 15 seconds. Incubate at room temperature for 2 minutes. (f) Centrifuge at 12,000 rpm for 10 minutes at 4°C. (g) Pipette the supernatant into a 96-well 2.2 mL deep-well plate.

[0116] The RNA sample preparation procedure is as follows. (a) 600 μL of binding buffer was added to columns 1 and 7. (b) 600 μL of DNase stop buffer was added to column 2 / 8. (c) 600 μL of wash buffer was added to columns 3, 4 and 9, 10. (d) 100 μL of elution buffer was added to column 5 / d 11. (e) 190 μL of DEPC water and 10 μL of MagaBio® reagent were added to columns 6 and 12. (f) Transfer 300 μL of the colorless upper phase containing RNA to columns 1 and 7. (g) Place the test plate in the nucleic acid purification device. (h) Set up the equipment as per Table 3 below.

[0117] [Table 3]

[0118] The eluate was transferred to a clean nuclease-free centrifuge tube and the RNA sample was stored at -80°C.

[0119] To use the Takara Prime Script™ Reverse Transcription System, mix each component and briefly centrifuge before use. Collect the volumes of the following components as shown in Table 4.

[0120] [Table 4]

[0121] (a) Keep on ice until adding the reverse transcription mix. Centrifuge for 10 seconds in a microcentrifuge. Heat in a 42°C heat block for 2 minutes and immediately chill in ice-water at 4°C for at least 5 minutes. (b) For each cDNA reaction, prepare reverse transcription reaction mix, 10 µL. Assemble on ice in the order listed. Volumes of components in step b) (Table 5).

[0122] [Table 5]

[0123] (c) Combine 20 μL of reverse transcription mix and centrifuge in a microcentrifuge for 10 seconds. (d) Anneal in heat block at 37° C. for 15 min, 85° C. for 5 s, and store at 4° C. on an S100™ thermal cycler. (e) Aliquot the cDNA sample and store it at -20°C.

[0124] QT-PCR procedure (a) Prepare qPCR reaction mix in one tube based on the number of assays and replicates. (b) Prepare the primer and master mix mixture according to the manufacturer's instructions and dispense 9 μL of the mixture into each well of a 384 qPCR plate. (c) Transfer 1 μL of cDNA to each well of the 384 qPCR plate already containing 9 μL of mixture. Briefly centrifuge the plate. (d) Seal the plate (e) Mix thoroughly and swirl gently. (f) Running the PCR reaction (g) Place the reaction plate into the instrument. (h) Set up cycling conditions on the Applied Biosystems ViiA™ 7 per Table 6.

[0125] [Table 6]

[0126] Real-time quantitative QT-PCR statistics

[0127] Data were expressed as mean ± SD. The 2-ΔΔCT method was used to analyze mRNA expression levels. Results were normalized using glyceraldehyde-3-phosphate dehydrogenase (GAPDH or other internal standard) for RNA input. Statistical analysis was performed using one-way ANOVA and post-hoc Dunnett's test in cases of significance. When N was very small or the data did not follow a Gaussian distribution, non-parametric tests such as the Mann-Whitney and Kruskal-Wallis tests were used. Differences were considered significant if p<0.05.

[0128] Histological analysis

[0129] H&E staining: Colon segments were cut to a thickness of 4 μm, dried in an oven for 1 h, and stained with H&E solution using our standard protocol. Briefly, segments were stained with hematoxylin solution for 90 s, followed by staining in eosin solution for 30 s. They were then dehydrated and coverslipped for later image analysis. H&E stained images were scanned using a Leica Aperio® GT450. The severity of inflammatory cell infiltration and tissue damage was semiquantitatively assessed according to the pathology score criteria in Table 7.

[0130] [Table 7]

[0131] PAS staining: Colon sections were cut to a thickness of 4 μm, dried in an oven for 1 h, and stained with the PAS kit (Gefan#M035) using our standard protocol. Briefly, sections were stained with periodic acid reaction solution for 8 min. Then washed 3 times with distilled water for 5 min. Stained in Schiff's solution for 10 min. Transformed into distilled water 3 times for 5 min. Then dehydrated and coverslipped for later image analysis.

[0132] The total length of the colon was measured with a ruler and then divided into three parts, and the central 100 microns of each part was taken for goblet cell count. The number of goblet cells was calculated for each 100 microns of colonic tissue, and then the average number of goblet cells from the three parts was used for further analysis.

[0133] 4. Data Analysis

[0134] In vivo animal data were expressed as mean ± SEM. Statistical analysis was performed by one-way ANOVA followed by Dunnett's multiple comparisons. When N was very small or the data did not follow a Gaussian distribution, non-parametric tests such as Whitney were used. Differences were considered significant when p<0.05.

[0135] result

[0136] Weight and weight change

[0137] As shown in Figure 2, body weight did not show significant differences in all groups, but as shown in Figure 3, body weight change was significantly decreased on day 2 after 3.61 mg / kg lobeglitazone treatment and on days 2, 3, 4, 5, and 6 after 30 mg / kg CSA treatment compared to control (no drug) group mice. There was no significant body weight change after 1.20 mg / kg and 0.36 mg / kg lobeglitazone treatment compared to control group mice.

[0138] DAI Score

[0139] As shown in Figure 4, DSS treatment significantly increased mouse DAI on days 7, 8, 9, and 10. Compared to the control group, treatment with 30 mg / kg CSA significantly decreased mouse DAI on days 8, 9, and 10. Treatment with lobeglitazone did not significantly reduce the DAI scores of the animals.

[0140] Colon length and colon weight

[0141] As shown in Figure 5, DSS treatment did not significantly change colon weight. Compared to the control group, treatment with 30 mg / kg CSA and lobeglitazone did not significantly change colon weight. As shown in Figure 6, DSS treatment significantly shortened colon length. Compared to the control group, treatment with 30 mg / kg CSA significantly extended colon length. Treatment with lobeglitazone did not significantly extend colon length. As shown in Figure 7, DSS treatment significantly increased colon length / weight ratio. Compared to the control group, treatment with 30 mg / kg CSA significantly decreased colon length / weight ratio. Treatment with lobeglitazone significantly decreased colon length / weight ratio.

[0142] Gene expression levels in the colon

[0143] As shown in Figure 8, the gene expression level of TNF-α was significantly increased in the colon after DSS treatment compared to the untreated group. Treatment with lobeglitazone (0.36 and 1.20 mg / kg) significantly decreased the gene expression level of TNF-α. Treatment with 30 mg / kg CSA decreased TNF-α expression. Treatment with lobeglitazone (3.61 mg / kg) did not show any obvious change in the gene expression level of TNF-α.

[0144] As shown in Figure 9, the gene expression level of IL-10 is decreased after DSS treatment compared to the untreated group, which did not change significantly after lobeglitazone and 30 mg / kg CSA treatment.

[0145] As shown in Figure 10, the gene expression level of CCL2 was significantly increased in the colon after DSS treatment compared to the untreated group. The gene expression level of CCL2 was significantly decreased after 3.61 mg / kg lobeglitazone and 30 mg / kg CSA treatment. Treatment with lobeglitazone (0.36 and 1.20 mg / kg) resulted in a decrease in the level of CCL2 gene expression.

[0146] As shown in Figure 11, the gene expression level of IL-1β was significantly increased in the colon after DSS treatment compared to the untreated group. The gene expression level of IL-1β was significantly decreased after treatment with the compounds lobeglitazone (0.36 and 1.20 mg / kg) and 30 mg / kg CSA, but the level of IL-1β gene expression was unchanged after treatment with 3.61 mg / kg lobeglitazone.

[0147] Colon H&E staining and scoring

[0148] The untreated group showed intact mucosal structure and many goblet cells, while the 5% DSS-induced model group showed severe inflammatory cell infiltration, glandular separation, crypt loss and damage to mucosal structure, and significantly increased the H&E total score. Treatment with 30 mg / kg CSA significantly decreased the H&E total score. Treatment with lobeglitazone (0.36, 1.20 and 3.61 mg / kg) resulted in a decrease in the H&E total score (Figure 12).

[0149] Compared with the untreated group, goblet cells were significantly decreased in the DSS-treated group. Goblet cells were significantly increased after 30 mg / kg CSA treatment. Treatment with compound lobeglitazone (0.36 and 1.20 mg / kg) tended to increase the number of goblet cells, and treatment with 3.61 mg / kg lobeglitazone did not change the number of goblet cells (Figure 13).

[0150] conclusion

[0151] Lobeglitazone was evaluated in a mouse IBD model (DSS model) at 0.36, 1.20, and 3.61 mg / kg. Treatment with lobeglitazone was shown to significantly reduce GI inflammation as evidenced by gene expression data. Lobeglitazone did not significantly reduce DAI score and colon weight / length ratio.

[0152] Example 2: M1 macrophage differentiation assay procedure

[0153] The objective of this study was to evaluate the effects of lobeglitazone and other thiazolidinediones on M1 macrophage differentiation.The test articles and vehicles used in this study were lobeglitazone sulfate, pioglitazone HCl, and rosiglitazone.

[0154] Reconstitution of media and solutions: (1) Prepare complete media: Add 50 mL of HI FBS (10%) and 5 mL of 100x penicillin / streptomycin (1%) to a 500 mL bottle of RPMI-1640; (2) Reconstitute LPS in sterile water to a stock concentration of 1 mg / ml; (3) Reconstitute human IL-4 by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (4) Reconstitute human M-CSF by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (5) Reconstitute human GM-CSF by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (6) Reconstitute test articles (M107, pioglitazone, rosiglitazone) in DMSO to a stock concentration of 10 mM.

[0155] The timeline for the M1 macrophage differentiation assay was as shown in Table 8.

[0156] [Table 8]

[0157] Drug concentrations tested in the M1 differentiation assay are shown in Table 9.

[0158] [Table 9]

[0159] Experimental procedure (1) Reactivate frozen monocytes. (2) Count the cells and adjust the cell density to 0.5 × 10 cells in fresh complete differentiation medium (RPMI1640 + 10% FBS + 1% PS supplemented with 50 ng / ml GM-CSF or M-CSF). 6 Adjust to pieces / ml. (3) Seed the isolated monocytes at 100 μL per well in a 96-well plate. (4) Using the D300e, add the test substances according to the plate map (final concentrations in μM). (5) Differentiate monocytes into macrophages, M1 in the presence of GM-CSF, or M2 with M-CSF, for 5 days. (6) On day 5, cytokines are added at twice the final concentration. (6a) Prepare medium containing GM-CSF for M1 and M-CSF for M2. (6b) Add the corresponding activator to achieve 2x the final reaction concentration: (i) IFNg stock 0.2 mg / ml: for 100 ng / ml, add 2.5 μL of stock to 5 ml of full differentiation medium; (ii) LPS stock 1.0 mg / ml: dilute to 20 μg / ml by mixing 2 μL of stock into 100 μL of medium; then for 40 ng / ml, add 10 μL of 20 μg / ml LPS to 5 ml of full differentiation medium; (iii) IL-4 stock 0.1 mg / ml: for 40 ng / ml, dilute 2 μL of stock into 5 ml of full differentiation medium. (6c) Add 100 μL of cytokine-containing differentiation medium to each well, for a total volume of 200 μL in each well. (6d) Add test compound again using the D300e, making sure to set the final volume to 200 μL. (7) The cytokines and test compounds are incubated for 48 hours. (8) On day 7, spin down plates at 300×g for 5 min and collect supernatants using MSD for cytokine quantification. (9) M1-associated macrophage cytokines TNF-α, IL-1b, IL-6, IL-12, and IL-23 were quantified.

[0160] result

[0161] The results of the M1 macrophage cytokine expression assay are shown in Figure 14. The results of the M1-like macrophage cytokine expression assay are shown in Figures 14 to 18.

[0162] conclusion

[0163] This study evaluated the effects of lobeglitazone (M107), pioglitazone (pigl), and rosiglitazone (Rosig) on ​​the differentiation of M1 and M2 macrophages using cytokine assays. Among the three test substances, pioglitazone and rosiglitazone promoted M1 macrophage cytokines in a dose-dependent manner, whereas lobeglitazone showed no promotion of M1 macrophage cytokines.

[0164] Example 3: M2 macrophage differentiation assay procedure

[0165] The objective of this study was to evaluate the effects of lobeglitazone and other thiazolidinediones on M2 macrophage differentiation.The test articles and vehicles used in this study were lobeglitazone sulfate, pioglitazone HCl, and rosiglitazone.

[0166] Reconstitution of media and solutions: (1) Prepare complete media: Add 50 mL of HI FBS (10%) and 5 mL of 100x penicillin / streptomycin (1%) to a 500 mL bottle of RPMI-1640; (2) Reconstitute LPS in sterile water to a stock concentration of 1 mg / ml; (3) Reconstitute human IL-4 by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (4) Reconstitute human M-CSF by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (5) Reconstitute human GM-CSF by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (6) Reconstitute test articles (M107, pioglitazone, rosiglitazone) in DMSO to a stock concentration of 10 mM.

[0167] The timeline for the M2 macrophage differentiation assay was as shown in Table 10.

[0168] [Table 10]

[0169] The drug concentrations tested in the M2 differentiation assay are shown in Table 11.

[0170] [Table 11]

[0171] Experimental Procedures (Monocyte Isolation, Treatment and Plating) (1) Isolate monocytes from PBMCs using the StemCell® Monocyte Isolation Kit (2) Count the cells and adjust the cell density to 0.5 × 10 cells in fresh complete differentiation medium. 6 Adjust to cells / ml. Add 5 μL of M-CSF stock (final concentration of 100 μg / ml) to each 5 ml cell mixture. (3) Seed the isolated monocytes at 100 μL per well in a 96-well plate. (4) Add test articles according to plate map using D300e (final concentration in μM). (5) Differentiate monocytes into macrophages over a 4-day period and into M4 cells in the presence of M-CSF. (6) On day 4, additional medium containing M-CSF is added. (7) Add test compound using the D300e (final volume = 100 μL). (8) On day 6, add IL-4 to 5x final concentration (IL-4 stock 0.1 mg / ml: for 100 ng / ml, dilute 10 μL of stock into 10 ml of medium. Add 50 μL of 100 ng / ml IL-4 to each well containing 200 μL of medium to give 20 ng / ml). (9) On day 8, spin down plates at 300×g for 5 min and collect supernatants using MSD for cytokine quantification. (10) M2-associated macrophage cytokines of interest are listed in Table 12 below.

[0172] [Table 12]

[0173] (11) Harvest cells for flow analysis of viability / CD11c / CD163 / CD206.

[0174] result

[0175] Flow cytometry and surface markers CD163, CD11c, and CD206 are applied to characterize M2 repolarization. Unstained and stained with surface marker antibodies all confirm optimization of the gating strategy.

[0176] Detailed cell phenotypes are shown in FIG. 19, FIG. 20 and FIG.

[0177] Cytokine and chemokine expression was quantified using MSD. Figure 22 and Figure 23 show the overall M1- and M2-associated cytokine expression, respectively. The detailed M2-associated cytokines MDC, Eotaxin-2, and IL-1RA are shown in Figure 24, Figure 25, and Figure 26.

[0178] conclusion

[0179] This study evaluated the effects of lobeglitazone, pioglitazone, and rosiglitazone on M2 macrophage differentiation using flow cytometry and cytokine assays. As shown by flow cytometry studies, M2 macrophage surface marker characterization demonstrated that M107 promoted M2 repolarization at lower concentrations (~0.025 μM) compared to pioglitazone HCl (~10 μM) and rosiglitazone (~0.4 μM) in donors 1 and 2. Cytokine analysis also showed a significant promotion of MDC expression (in donor 3) and eotaxin-2 expression (in donor 1).

[0180] Example 4: M1 to M2 macrophage repolarization assay procedure

[0181] The objective of this study was to evaluate the effects of lobeglitazone and other thiazolidinediones on the repolarization of M1 to M2 macrophages.The test articles and vehicles used in this study were lobeglitazone sulfate, pioglitazone HCl, and rosiglitazone.

[0182] Reconstitution of media and solutions: (1) Prepare complete media: add 50 mL of HI FBS (10%) and 5 mL of 100x penicillin / streptomycin (1%) to a 500 mL bottle of RPMI-1640; (2) Reconstitute LPS in sterile water to a stock concentration of 1 mg / ml; (3) Reconstitute hIL-4 in sterile water to a stock concentration of 100 μg / ml; (4) Reconstitute human M-CSF by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (5) Reconstitute human GM-CSF by solubilizing in sterile water to 0.1 mg / ml by pipetting the mixture; (6) Reconstitute test articles (M107, pioglitazone, rosiglitazone) in DMSO to a stock concentration of 10 mM.

[0183] The timeline for the M1 to M2 macrophage repolarization assay was as shown in Table 13.

[0184] [Table 13]

[0185] The drug concentrations tested in the M1 differentiation assay are shown in Table 14.

[0186] [Table 14]

[0187] Experimental Procedures (Monocyte Isolation, Treatment and Plating) (1) Isolate monocytes from PBMCs using the StemCell® Monocyte Isolation Kit. (2) Count the cells and adjust the cell density to 0.5 × 10 cells in fresh complete differentiation medium. 6 Adjust to cells / ml (add 10 μL of M-CSF stock (100 ug / ml) to 10 ml of cell mixture for a final concentration of 50 ng / ml). (3) Seed the isolated monocytes at 100 μL per well in a 96-well plate. (4) On the fourth day, add 100 ul of medium containing M-CSF. (5) On day 6, stimuli are added at 5x final concentration (200 μL of medium per well) to give a final concentration of 50 ng / ml. (6) On day 8, the medium is removed and replenished with 200 μL of cytokine-free medium, after which test compounds (final concentration in μM) are added and incubated for 4 days. (7) On day 12, add IL-4 and incubate for 2 days (50 μL of 100 ng / ml IL-4 is added to each well containing 200 μL of medium to give 20 ng / ml). (8) On day 14, spin down plates at 300×g for 5 minutes and collect supernatants for cytokine quantification. (9) The M1-associated macrophage cytokines and M2-associated macrophage cytokines of interest are as shown in Table 15.

[0188] [Table 15]

[0189] Cells are then harvested for flow analysis of viability / CD11c / CD163 / CD206.

[0190] result

[0191] Cell surface markers CD163, CD11c, and CD206 are applied to characterize M2 repolarization via flow cytometry. To confirm the optimization of the gating strategy, controls include no staining and staining with all surface marker antibodies. Detailed cell phenotypes are shown in Figure 27.

[0192] Cytokine and chemokine expression was quantified using MSD. Figures 28 and 29 show the overall M1- and M2-associated cytokine expression, respectively. The detailed M2-associated cytokines MDC, eotaxin-2, and IL-1RA are shown in Figures 30-32.

[0193] conclusion

[0194] This study evaluated the effects of lobeglitazone, pioglitazone, and rosiglitazone on M1 to M2 repolarization using flow cytometry and cytokine assays. As shown by flow cytometry studies, M2 macrophage surface marker characterization showed that M107 promoted M1 to M2 repolarization at concentrations as low as approximately 0.05 μM. Cytokine analysis showed that M107 also showed a significant promotion of MDC expression at 0.1 μM, in contrast to pioglitazone and rosiglitazone at 5 μM and 6.4 μM, respectively.

[0195] Unless otherwise indicated, all numbers expressing properties such as amounts of ingredients, molecular weights, reaction conditions, and the like used in the specification and claims should be understood in all instances to be modified by the term "about." As used herein, the terms "about" and "approximately" mean within 10-15%, preferably within 5-10%. Thus, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations and may vary depending upon the desired properties sought to be obtained by the present invention. At the very least, and without intending to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0196] The terms "a," "an," "the," and similar reference words used in the context of describing the present invention (particularly in the context of the claims below) should be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value within the range. Unless otherwise indicated herein, each separate value is incorporated herein as if individually set forth herein. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples or representative language (e.g., "such as") provided herein is merely to better illustrate the present invention and does not otherwise limit the scope of the claimed invention. No term in this specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0197] Grouping of alternative elements or embodiments of the invention disclosed herein should not be construed as limiting. The members of each group may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. It is anticipated that for reasons of convenience and / or patentability, one or more members of a group may be included in or excluded from a group. When any such inclusion or exclusion occurs, the specification shall be deemed to include the modified group so fulfilling all Markush group descriptions used in the appended claims.

[0198] Certain embodiments of the present invention, including the best mode known to the inventors for carrying out the invention, have been described herein. Of course, variations on these described embodiments will become apparent to those skilled in the art upon reading the foregoing description. The inventors expect those skilled in the art to use such variations as appropriate, and the inventors intend the invention to be carried out other than as specifically described herein. Accordingly, the present invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed herein unless otherwise indicated herein or otherwise clearly contradicted by context.

[0199] Certain embodiments disclosed herein may be further limited in the claims using the terms "consisting of" or "consisting essentially of." When used in the claims, whether as filed or added by amendment, the transition term "consisting of" excludes any element, step, or ingredient not specified in the claim. The transition term "consisting essentially of" limits the scope of the claim to those materials or steps specified and which do not materially affect the basic and novel characteristics. Embodiments of the invention so claimed are essentially or explicitly described and operable herein.

[0200] Additionally, throughout this specification, numerous references are made to patents and printed publications. Each of the above cited references and printed publications is individually incorporated herein by reference in its entirety.

[0201] Finally, it should be understood that the embodiments of the invention disclosed herein are illustrative of the principles of the invention. Other variations that may be employed are within the scope of the invention. Thus, by way of example, but not of limitation, alternative configurations of the invention may be utilized in accordance with the teachings herein. Thus, the invention is not limited to exactly as shown and described.

Claims

1. A method for protecting the digestive system from damage, comprising contacting cells of the digestive tract (GI) system with an effective amount of thiazolidinedione selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone.

2. The method according to claim 1, wherein the thiazolidinedione is robeglitazone.

3. The method according to claim 1, wherein the cells are derived from the mouth, pharynx (throat), esophagus, stomach, small intestine, large intestine, rectum, anus, salivary gland, liver, gallbladder, or pancreas.

4. The method according to claim 1, wherein the GI-type cells are absorptive cells (intestinal epithelial cells), goblet cells, pancreatic islet cells, gastrointestinal endocrine cells, hepatocytes, Paneth cells, fenestrated hepatic endothelial cells, Kupffer cells, serous cells, gastric chief cells, mucinous cells, smooth muscle cells, parietal cells, myoepithelial cells, stem cells, gastric surface mucinous cells, pancreatic acinar cells, taste buds, intercalated cells of Cajal (ICCs), or nerve cells.

5. The method according to any one of claims 1 to 4, wherein the cells are mammalian cells.

6. The method according to claim 5, wherein the cells are human cells.

7. The method according to any one of claims 1 to 4, wherein the cells are treated in vitro.

8. The method according to any one of claims 1 to 4, wherein the cells are treated ex vivo.

9. The method according to any one of claims 1 to 4, wherein the cells are treated in vivo.

10. The method according to any one of claims 1 to 4, wherein the cells have a GI system disease or disorder, or the subject is at risk of a GI system disease or disorder.

11. The method according to claim 10, wherein the GI system disease or disorder is selected from abdominal adhesions, adult acid reflux (gastroesophageal reflux disease or GERD), pediatric acid reflux (GERD), anatomical problems of the lower GI duct, appendicitis, Barrett's esophagus, bowel management problems (fecal incontinence), celiac disease, colon polyps, constipation, Crohn's disease, cyclic vomiting syndrome, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, pseudo-obstruction, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcer (gastric ulcer), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, and Zollinger-Ellison syndrome, or symptoms associated therewith.

12. The method according to claim 11, wherein the GI-related disease or disorder is gastroparesis.

13. The method according to claim 12, wherein the gastroparesis is idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis, or drug-induced gastroparesis.

14. A method for treating a GI system disease or disorder in a subject requiring such treatment, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a thiazolidinedione selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone.

15. The method according to claim 14, wherein the thiazolidinedione is robeglitazone.

16. The method according to claim 14, wherein the GI system disease or disorder is selected from abdominal adhesions, adult acid reflux (GERD), pediatric acid reflux (GERD), anatomical problems of the lower GI duct, appendicitis, Barrett's esophagus, bowel management problems (fecal incontinence), celiac disease, colon polyps, constipation, Crohn's disease, cyclic vomiting syndrome, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, indigestion (dyspepsia), inguinal hernia, pseudo-obstruction, IBD, IBS, lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcer (gastric ulcer), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, and Zollinger-Ellison syndrome, or symptoms associated therewith.

17. The method according to claim 16, wherein the GI-related disease or disorder is gastroparesis.

18. The method according to claim 17, wherein the gastroparesis is idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis, postoperative gastroparesis, or drug-induced gastroparesis.

19. The method according to any one of claims 14 to 18, wherein the subject is a mammal.

20. The method according to claim 19, wherein the mammal is a non-human mammal.

21. The method according to claim 19, wherein the mammal is a human.

22. The method according to any one of claims 14 to 18, wherein the thiazolidinedione is administered in a dose of 0.01 mg or more.

23. The method according to any one of claims 14 to 18, wherein the thiazolidinedione is administered in a dose of 0.01 to 5000 mg / day.

24. The method according to any one of claims 14 to 18, wherein the thiazolidinedione is administered parenterally, enterally, topically, orally, sublingually, orally, intrapulmonaryly, intranasally, intramuscularly, or subcutaneously.

25. Use of thiazolidinediones selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone, to alleviate the signs and symptoms of gastric paresis.

26. The use according to claim 25, wherein the severity of the signs and symptoms of gastroparesis is measured by the Gastroparalysis Major Symptom Index Diary (GCSI-DD) of the American Society of Neurogastropology and Exercise.

27. The use according to claim 25, wherein the severity of the signs and symptoms of gastroparesis is measured by the change in the GCSI-DD score from baseline to 4 weeks of treatment, 8 weeks of treatment, 12 weeks of treatment, or 1 year of treatment.

28. Use of thiazolidinediones selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone, to reduce the severity of nausea in subjects with gastroparesis.

29. Use of thiazolidinediones selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone, to reduce the severity of early satiety in subjects with gastroparesis.

30. Use of thiazolidinediones selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone, to reduce the severity of postprandial bloating in subjects with gastroparesis.

31. Use of thiazolidinediones selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone, to reduce the severity of upper abdominal pain in subjects with gastroparesis.

32. Use of thiazolidinedione selected from one or more of pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone to reduce the severity or number of vomiting cases in subjects with gastric paresis.

33. Use of thiazolidinediones selected from one or more of the following to reduce the overall severity of gastroparesis: pioglitazone, rosiglitazone, lobeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone.

34. Use of thiazolidinediones selected from one or more of pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone to reduce GI inflammation, as suggested by a decrease in the expression of one or more of the TNF-α, IL-10, CCL2, and IL-1β genes in target cells experiencing GI inflammation.

35. Use of thiazolidinediones selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone, to delay, halt, or reverse the progression of gastric paresis.

36. The use according to any one of claims 25 to 35, wherein the thiazolidinedione is lobeglitazone.

37. A method for treating GI disease in a mammal, comprising administering to the mammal an effective amount of thiazolidinedione selected from one or more of the following: pioglitazone, rosiglitazone, robeglitazone, siglitazone, dalglitazone, deuterium-stabilized R-pioglitazone, englitazone, religlitazone, netoglitazone, riboglitazone, troglitazone, and paraglitazone.

38. The method according to claim 37, wherein the GI disease is gastroparesis.

39. The method according to claim 38, wherein the gastroparesis is idiopathic gastroparesis, diabetic gastroparesis, postoperative gastroparesis, or drug-induced gastroparesis.

40. The method according to any one of claims 37 to 39, wherein the thiazolidinedione is lobeglitazone.

41. A method for inhibiting M1 macrophage differentiation in an individual, comprising administering an effective amount of lobeglitazone to the individual.

42. The method according to claim 41, wherein a GI system disorder is treated in the individual as a result of the inhibition of M1 macrophage differentiation.

43. A method for promoting M1 macrophage differentiation in an individual, comprising administering an effective amount of pioglitazone or rosiglitazone to the individual.

44. A method for promoting M2 macrophage differentiation in an individual, comprising administering an effective amount of robeglitazone, pioglitazone, or rosiglitazone to the individual.

45. The method according to claim 44, wherein, as a result of the promotion of M2 macrophage differentiation, a GI system disorder is treated in the individual.

46. A method for promoting the repolarization of M1 macrophages to M2 macrophages in an individual, comprising administering an effective amount of robeglitazone, pioglitazone, or rosiglitazone to the individual.

47. The method according to claim 46, wherein a GI system disorder is treated in the individual as a result of the aforementioned promotion of repolarization from M1 macrophages to M2 macrophages.

48. The method according to claim 44 or 46, wherein the thiazolidinedione is robeglitazone.

49. The method according to claim 45 or 47, wherein the GI system disorder is selected from abdominal adhesions, adult acid reflux (GERD), pediatric acid reflux (GERD), anatomical problems of the lower GI duct, appendicitis, Barrett's esophagus, bowel management problems (fecal incontinence), celiac disease, colon polyps, constipation, Crohn's disease, cyclic vomiting syndrome, diarrhea, diverticulosis and diverticulitis, dumping syndrome, food poisoning, gallstones, gas, gastritis, gastroparesis, GI bleeding, hemorrhoids, dyspepsia, inguinal hernia, pseudo-obstruction, IBD, IBS, lactose intolerance, liver disease, microscopic colitis, intestinal stoma surgery, pancreatitis, peptic ulcer (gastric ulcer), proctitis, short bowel syndrome, ulcerative colitis, viral gastroenteritis, and Zollinger-Ellison syndrome, or symptoms associated therewith.

Images